Compounds inhibiting (blocking) bitter taste, methods for use and production thereof


FIELD: chemistry.

SUBSTANCE: invention relates to a compound of general formula (I) or pharmaceutically acceptable salts thereof, where Alk is an C1-C6alkyl group; G is C=O and Q is CR51R52 or NR51, where R51 and R52, being identical or different, independently denote H, C1-C6alkyl, optionally substituted with a substitute selected from a group comprising carboxy, phenoxy, benzyloxy, C1-C6alkoxy or hydroxy; C3-C6cycloalkylC1-C6alkyl; phenylC1-C6alkyl, optionally substituted with a halogen; phenylamidoC1-C6alkyl; phenylC1-C6alkylamidoC1-C6alkyl, optionally substituted with a C1-C6alkoxy group; or R51 and R52, together with a carbon atom with which they are bonded form a C=O or C2-C6alkenyl group, optionally substituted with a phenyl; M1 is CR49, where R49 is H; M2 is CR50, where R50 is H; R38 is H, C1-C6alkyl, substituted with a phenoxy group; C3-C6cycloalkylC1-C6alkyl; arylC1-C6alkyl, optionally substituted with 1 or 2 substitutes selected from a group comprising C1-C6alkyl, C1-C6alkoxy, C1-C6alkoxycarbonyl, carboxyl, N-methylamido, hydroxy, C1-C6alkoxyC1-C6alkoxy, C1-C6alkylthio, C1-C6alkylsulphanyl, cyano, halogen, perfluoroC1-C6alkyl, nitro, formyl, hydroxyC1-C6alkyl and amino, wherein the aryl moiety is a phenyl or naphthyl; and heteroarylC1-C6alkyl, where the heteroaryl moiety is pyridinyl, optionally substituted with 1 or 2 groups selected from C1-C6alkoxy or hydroxyC1-C6alkyl, pyrazolyl or isoxazolyl, substitute with 1 or 2 C1-C6alkyl groups; R47 and R48 is C1-C6alkyl. The invention also relates to specific compounds, a method of reducing or weakening bitter taste, a composition of a food/non-food product or beverage or drug for reducing or lightening bitter taste and a method of producing a compound of formula (I).

EFFECT: obtaining novel compounds which are useful as bitter taste inhibitors or taste modulators.

37 cl, 6 dwg, 12 tbl, 186 ex

 

The present application claims priority under provisional patent application U.S. ser. No. 60/957129, filed August 21, 2007, and provisional application U.S. ser. No. 61/047187, filed April 23, 2008, and refers to the application of U.S. ser. No. 11/766974 representing a partial continuation of application U.S. ser. No. 11/555617, filed November 1, 2006, which, in turn, is a partial continuation of application U.S. ser. No. 10/191058, filed July 10, 2002, and also is a partial continuation of application U.S. ser. No. 10/742209, filed December 1, 2003, which is a divisional application of U.S. ser. No. 09/825882 filed April 5, 2001, now U.S. patent No. 7105650, all of these applications are included in this text in full by reference.

The technical FIELD

The present application relates to the identification of gustatory receptors in human type 2 (hT2R) and their application in analysis tools to identify ligands that activate specific T2R. In previous patent applications of the present authors, they described the functional expression of bitter taste receptors in humans, including hT2R8 and hT2R14. In this application it is reported that hT2R8 and hT2R14 activated fraction coffee enriched with bitter compounds. Also reported the identification of antagonists of hT2R8 and hT2R14 using high-performance screening analysis, and that the combination is of antagonists can reduce the bitter taste of coffee and fractions coffee. According to the present invention, a method of changing and improving the taste of the coffee drinks.

In particular, according to the present invention, the application of hT2R8 and/or hT2R14 in the screening analysis and taste samples to identificatie compounds inhibiting (blocking) the bitter taste of coffee and other beverages.

Also, the present invention relates to the identification of the ligand with the antagonist properties of bitter taste, broad-spectrum, i.e. ligand, which would have the ability to substantially block or inhibit activation of multiple (13) bitter taste receptors a number of different ligands bitter taste, and to block or inhibit the activation of six other bitter taste receptors and to inhibit bitter taste caused some bitter compounds, which to date have not identified receptor(s) bitter taste.

In particular, the invention relates to the identification of the ligand specified in the present description as the Connection with the antagonist properties of bitter taste, broad-spectrum, i.e. substantially blocking or inhibiting activation hT2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 64, 65 and 71 of bitter taste receptors variety of bitter ligands, and blocking or inhibiting the activation of the other six Retz is Perov bitter taste, i.e. hT2R5, 9, 13, 54, 67 and 75, as well as inhibiting the bitter taste caused some bitter compounds, which to date have not identified receptor(s) bitter taste.

Also, in particular, in accordance with the present invention found that such a connection antagonist reduces the bitter taste of salicin, antagonist hT2R16, and phenyltoloxamine, agonist hT2R51.

Also, in particular, according to the present invention, the detected block the specified connection-antagonist bitter taste caused by bitter compounds that activate different receptors, bitter taste, including omeprazole, activating hT2R10, 14 and 75; Rebaudioside A, a natural sweetener, activating at least 7 of bitter taste receptors; and further suppression of the same antagonist bitter taste caused by bitter compounds, bitter taste receptors which are unknown, including dextromethorphan and diphenhydramine.

As described above, according to the present invention, the proposed use of such compounds in foods, beverages, medicines and other products consumption in order to reduce their bitter taste, including the bitter taste caused by unidentified bitter ligands or compounds, when the feeling of bitterness is caused by activation of multiple receptors is of Oriago taste, or when its specificity to the receptor is not installed.

Also, on the basis described above, according to the present invention proposed the use of this antagonist in order to identify conservative motif presented in different T2R person involved in ligand binding, and activation of T2R and construction of chimeric and mutant receptors associated with G-protein (GPCR), designed in such a way that they contain the specified motif.

Further, the present invention relates to any of the compounds identified using the described screening analysis, and its application in foods, beverages and medicines, including coffee and foods with added coffee and drinks and medicines.

The LEVEL of TECHNOLOGY

One of the basic tastes that are recognized by the person, is bitter. Until recently, physiology bitter taste was very poorly understood. Recently started research to clarify the biology of taste (Lindemann, Nature (2001)). It is now known that many bitter compounds cause a bitter taste due to interaction with receptors on the cell surface. These receptors belong to the family of receptors consists of seven transmembrane domains, which interact with intracellular G-proteins. the humans and rodents has been allocated a new family of GPCR, named T2R (Adler et al., Cell 100(6):693-702 (2000); Chandrashekar et al., Cell 100(6): 703-711 (2000); Matsunami H, Montmayeur J P, Buck L B. Nature 404(6778): 601-4 (2000)). According to the claimed invention offers a number of evidence that T2R are mediators of response to bitter compounds. First, T2R genes specific manner expressed in subpopulations of cells gustatory receptor language and epithelium of the sky. Secondly, the gene of one of the T2R person (hT2R1) is the locus of a chromosome that is associated with sensitivity to the bitter compound 6-n-propyl-2-thiouracil in humans (Adler et al., (Id) (2000)). Thirdly, one of the T2R mouse (mT2R5) is the locus of a chromosome that is associated with sensitivity to the bitter compound to cycloheximide in mice. It was also shown that mT2R5 can activate gustducin, G-protein-specific manner expressed in taste cells and is associated with the transformation of the bitter stimulus (Wong et al., Nature 381:796-800 (1996)). Activation of gustducin mT2R5 occurs only in response to cycloheximide (Chandrashekar et al., (Id.) (2000). Thus, an assumption was made that the family mT2R is the mediators of the response to bitter taste in mice, while the family hT2R are mediators of response to bitter taste in humans. Only T2R suggested the presence of identified bitter ligand. It was shown that hT2R4 activated by denatonium (Chandrashekar et al., (Id.) 2000). Although effective to the ncentratio denatonium, used in research (1.5 mm), were unusually high, i.e. in 105times higher than okazavshayasya earlier boundary perception of bitter for denatonium in humans (Saroli, Sciences) 71:428-429 (1984)). Thus, not detected specific bitter ligands, fully compatible with any of the hT2R. It was also made the assumption that each of the hT2R is able to bind a lot of bitter ligands. This assumption is based on the fact that the family hT2R consists of only 25 identified receptors, while people can identify hundreds of different compounds as bitter. Sequence hT2R were previously published and are disclosed in published PCT applications Zuker et al. (WO 01/18050 A2, (2001)) and Adler et al. (WO 01/77676 A1 (2001)), both applications incorporated into the present text in full by reference.

One of the difficulties in the study of functions T2R is that these receptors cannot easily Express in cultured cell lines mammals. To improve the expression T2R to sequences T2R was added N-terminal sequence of well-expressed GPCR, rhodopsin (Chandrashekar et al., (Id.) 2000). This N-terminal fragment was also possible to easily control the expression of protein using available antibodies. Additionally, to improve the expression T2R used the SSTR3 terminal fragment (Bufe et al., Nat. Genet. 32:397-400 (2002)), the other N-terminal fragment. Despite the fact that the introduction of adoptinga terminal fragments were improved T2R expression of some cell lines mammals, many of them not expressibility sufficient for research functions. When using other methods, mT2R5 successfully expressed in insect cells Sf9, and they were used in the research functions using biochemical analysis of the binding of GTPγS (Chandrashekar et al., (Id.) 2000).

In my earlier filed application U.S. ser. No. 09/825882, now U.S. patent No. 7105650, the applicants of the present patent application identified and presented to the nucleic acid sequences and sequence of the polypeptides of several new at the time of publication of taste receptors of human rights, including hT2R51, hT2R54, hT2R55, hT2R61, hT2R63, hT2R64, hT2R65, hT2R67, hT2R71, and hT2R75. In addition, in patent applications U.S. ser. No. 11/182942and 10/628464 included in the present description in its entirety by reference, applicants represent sequences of polypeptides and DNA of another selected new taste receptor, named hT2R76.

Also, in the patent application U.S. ser. No. 10/191058 fully incorporated into the present text by reference, applicants have identified ligands specific way of activating three different T2R person. In addition, applicants recently applied for a U.S. patent ser. No. 11/455693, placed in this text in its entirety by reference, which further identify the bitter ligands specific way communicating with other T2R person, and provide the appropriate tests.

Also, in regards to the practical implementation of the present invention, were published evidence that both types of taste receptors: T2R and T1R are expressed in the gastrointestinal tract. For example, Wu et al., Proc, Natl. Acad. Sci USA 99(4):2392-7(2002) reported that T2R expressed in endocrine cells of the gastrointestinal tract (cells STC1), as well as subunits of gustducin and transducin, and that these cells probably respond to bitter ligands in the gastrointestinal tract. Also, Chen et al., AM J. Physiol. Cell Phyisol. 291(4):C726-39 (2006) reported that these incentives bitter taste induce the transfer of Vignola with the participation of Ca++ and the release of cholecystokinin (CCK) in endocrine cells of STC-1. Also, Rozengurt, A. J. Physiol. Gastrointes Liver Physiol. 291(2):G171-7 (2006) report that the taste receptors in the intestine, probably play a role in molecular sensitivity and control of digestive function and hormonal and/or neural pathways, and that they may play a role in the recognition of harmful drugs and the reactions of survival. Next, Sternini Am. J. Physiol. Gastrointest Liver Physiol. 292(2):G457-61 (2007) reported that taste receptors in the intestine does not necessarily participate in p is separately functions, such as molecular sensitivity, the absorption of nutritive compounds, protection from harmful compounds, and, further, suggested that the understanding of these mechanisms may be important in the treatment of these painful conditions like digestive disorders and inflammation. Further, recently, Mace et al., J. Physiol. 2007 (Epub) an assumption was made that T2R and T1R activate phospholipase C beta 2, PLC-beta2, and probably in the intestine there is a molecular sensor system similar to the system existing in the cells of the language, and these intestinal cells, as cells of the ciliated epithelium or dynamics of single cells expressing taste buds, can cause increased levels of GLUT2 and participate in the recognition of nutritional compounds, as well as in the regulation of nutrition in the treatment of obesity and diabetes. Also, Cui et al., Curr. Pharm. Des. 12(35):4591-600 (2006) suggest that the T1R expressed in the intestine, can be used in the study of compounds intended for the treatment of obesity and diabetes, as well as artificial sweeteners.

However, despite all the published data and the understanding that the receptors of the T2R family regulate the perception of bitter taste, and, optionally, participate in digestive function, there is a need to identify specific ligands, to enable the General taste buds human T2R. A better understanding of the binding capacity of different T2R, in particular T2R person, will be very useful because it will facilitate their use in the selection of compounds with the desired properties modulation of taste, i.e. blocking or inhibiting individual taste bitter compounds. Also, it will facilitate the identification of compounds suitable for the treatment and modulation of gastrointestinal function and related diseases such as obesity, diabetes, malabsorption of food, recognition of food, eating disorders, and in the regulation associated with the function of hormones and peptides, such as GLUT2, cholecystokin, and others

BRIEF description of the INVENTION

Accordingly, the present invention relates to detection of the fact that hT2R8 and hT2R14 enriched activated by bitter compounds faction coffee.

Also, according to the present invention the application of the invention to identify antagonists of hT2R8 and hT2R14, inhibiting or blocking the bitter taste of coffee and related to coffee products, beverages and medicines.

Further, the present invention relates to specific compounds-antagonists (blockers bitter taste), the overwhelming bitter taste of coffee and food, beverages and medicines with the addition of coffee.

Also, the present invention apply is to identify ligand, with the antagonist properties of bitter taste, broad-spectrum, i.e., it substantially blocks or inhibits the activation of many (13) bitter taste receptors variety of bitter ligands and blocks or inhibits the activation of six other bitter taste receptors, and inhibits bitterness caused some bitter compounds, the receptor(s) bitter taste which is not revealed in the present time.

In particular, the present invention relates to the identification of the ligand specified in the present description as compounds with antagonist properties bitter taste, broad-spectrum, i.e., it substantially blocks or inhibits the activation hT2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 64, 65 and 71 of bitter taste receptors variety of bitter ligands and blocks or inhibits the activation of six other bitter taste receptors, i.e. hT2R5, 9, 13, 54, 67 and 75, and also inhibits the bitterness caused some bitter compounds, the receptor(s) bitter taste which is not revealed in the present time.

Also, in particular, in accordance with the present invention found that such a connection antagonist reduces the bitter taste of salicin, antagonist hT2R16, and phenyltoloxamine, agonist hT2R51.

Also, in particular, according to the present invention discovered that the same connection-antag the NIST block bitter taste, called bitter compounds that activate multiple bitter taste receptors, including omeprazole, activating hT2R10, 14 and 75; Rebaudioside A, a natural sweetener, activating at least 7 receptor bitter taste; and that the same connection antagonist also inhibited bitter taste caused by bitter compounds that are not known the bitter taste receptors with which they interact, including dextromethorphan and diphenhydramine.

The present invention relates also to the use of compounds according to the present invention and related compounds in foods, beverages, drugs, or other means that provides the weakening of their bitter taste, including the bitter taste caused by unidentified bitter ligands or compounds, when the feeling of bitterness is caused by activation of multiple bitter taste receptors, or when its specificity to the receptor(s) is not installed.

Also, the present invention relates to food products, beverages and medicines containing at least one of the identified compounds antagonists Gorky, in a quantity sufficient to inhibit or block their bitter taste.

The facts on which is based the present invention, were found at the same time cell analysis, in which the measured activity T2R, using cells expressing a specific T2R in the presence or in the absence of specific ligands. In particular, as described below in the detailed description, in cell analysis to determine changes in the concentration of intracellular calcium, which used a cell line HEK expressing on its surface above specific T2R, then Express tropic chimeric G-protein, functionally associated with the specified T2R found that they are specific manner activates certain bitter compounds, while other hT2R in similar conditions is not active.

Thus, the present invention encompasses the use of the above receptors of the taste of a person means (methods) analysis, preferably in high-end analysis tools, to identify other compounds modulating, preferably blocking the activation of such receptors specified and other bitter compounds present in coffee and coffee related food and beverages.

Also, according to the present invention it is proposed to use these receptors to identify compounds that cause the sensation of bitter taste, in particular, present in coffee and food, beverages and medicines with doba is of coffee.

Also, according to the present invention it is proposed to use the connection-antagonist, has the properties of the antagonist bitter taste a wide spectrum of action, analysis tools, and taste samples of in vitro and in vivo to determine Gorky(s) connection(s) or bitter factions, for which this compound suppresses the sensation of bitter taste from drinking which inhibit this connection, and/or inhibits the activation of one or more bitter taste receptors specified bitter compound or fraction containing the specified bitter connection.

Further, in particular, according to the present invention it is proposed to use the specified connection with the antagonist properties of the bitter taste of a wide spectrum of action, in foods, beverages, medicines and other commodities intended for humans or animals, the intensity of the bitter taste which it is desirable to reduce.

The present invention also encompasses methods of analysis, including the additional step of assessing the impact of the identified modulating compounds for humans and other taste samples, and, in particular, the assessment of the impact of the identified compounds in bitter taste, in particular the bitter taste of coffee and fractions coffee containing one or more compounds that cause the sensation Gore is on taste.

Further, the present invention includes obtaining coffee and food, beverages and medicines with the addition of coffee, of which remove connection-specific way of activating the receptors bitter taste, for example, food and beverages processed with the removal or reduction of bitter compounds present in them.

According to some aspects of the present invention, also proposed structural classes of compounds, represented by the two following Frame Structures. On the Frame Structure 1 shows an example ourazalinova frame, and the Frame Structure 2 is an example of gigantinho frame.

Another specific object of the present invention is the use of the above compounds and analogues of structures 1 and 2 as inhibitors of the bitter-mediated receptor T2R8 applicable in the food/pharmaceutical sectors to mitigate the bitterness, in particular, coffee and food, beverages and medicines with the addition of coffee.

Another objective of the present invention is the evidence that established connections modulate, preferably inhibit, or block, the bitter taste in coffee and food, beverages and medicinal sredstvo coffee is added, using taste tests involving human or animal, preferably using taste tests involving human subjects. Example 1 illustrates the typical data sensitivity to one of these compounds. The data clearly show a significant decrease in bitterness for specific agonist T2R8 and a significant increase in efficiency in comparison with the known inhibitor T2R8.

Another objective of the present invention is the use of compounds according to the present invention as additives or flavor enhancers in compositions for inhibiting or blocking the bitter taste caused by compounds that are specific ways of activating receptors bitter taste. A preferred object of the invention is the use of compounds that inhibits activation of the receptor T2R8, to block the bitter taste of the compounds present in coffee and food, beverages and medicines with the addition of coffee.

Compounds identified according to the present invention, can be added to foods, beverages, cosmetics or medical compositions to modulate, preferably block bitter taste caused by activation of hT2R8 bitter compounds present in coffee and coffee related food products, beverages and medicines, Il is similar in structure compounds or other bitter compounds, for example, found in foods and beverages, or medicines and cosmetics, and causing the sensation of bitter taste.

OBJECTIVES of the INVENTION

The present invention is the presentation of research methods with the use of hT2R8 and/or hT2R14, and their chimeric variants and modifications determining compounds and compositions containing such compounds, causing or blocking the bitter taste of coffee and food, beverages and medicines with the addition of coffee.

The specific objective of the present invention is the provision of methods of analysis, to identify compounds that activate or blocking or modulating the activation and/or binding of hT2R8 with the compounds or compositions containing compounds are associated with a bitter taste of coffee.

Also a specific object of the present invention is the provision of methods of analysis, to identify compounds that activate or blocking or modulating the activation and/or binding hT2R14 with the compounds or compositions containing compounds are associated with a bitter taste of coffee.

Another specific object of the present invention is the provision of specific connections established using the methods of analysis according to the present from which retenu and compositions containing them, in particular, in the composition of coffee and food, beverages and medicines with the addition of coffee.

A specific object of the present invention is the provision of compounds below, which are antagonists T2R8 and T2R14 for which it was shown that they block the bitter taste of coffee.

Another specific object of the present invention is the use of compounds depicted above, as well as analogues Connection And Connection and Connection as inhibitors bitter taste to ease the bitter taste, mediated by receptors T2R8 and/or T2R14, food/pharmaceutical areas, particularly in coffee and food, beverages and medicines with the addition of coffee.

Another specific object of the present invention is the use of Connection and its analogues as inhibitors of the bitter taste of a wide range of actions to mitigate the sensation of bitter taste is mediated by any of the listed receptors person: T2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 64, 65 or 71 and/or T2R5, 9, 13, 54, 67 or 75, in the food/pharmaceutical fields, in particular in foods, beverages and medicines that contain a large number of bitter compounds, bitter compounds, interacting with multiple receptors bitter in the USA, or bitter compounds, the specificity of the receptors are not known.

Another specific object of the present invention is to provide compounds which can be represented by the following formulas.

According to the first aspect of the proposed compound of structural formula (I):

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar1represents a five - or six-membered aryl, heteroaryl or cycloalkyl ring;

m is 0, 1, 2 or 3;

R1represents the SO2; O=C; C=S; or C=NOR4;

X is selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R1' are independently selected from the group comprising hydrogen, halogen, ALK is l, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

or, alternatively, X and/or at least one of R1' together with the atoms to which they are linked, form an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cycloheptadiene or substituted cycloheptatriene ring, this ring may be fused with another aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cyclogeranyl or substituted cyclogeranyl ring;

R4-R8independently selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, zamestnavatelsky, heteroaryl, substituted heteroaryl, heteroaromatic and substituted heteroaromatic, or, alternatively, R5and R6, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring;

A and B are independently selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic; and

b is 0, 1 or 2.

According to some versions of the invention, A and B together with the nitrogen atom to which they are attached, form a ring which may be fused with additional substituted or unsubstituted rings and may contain at least one double bond. A non-limiting example of such a ring include a group having the formula:

According to the second aspect, the present invention proposed compounds of structural formula (II)below:

or a salt, hydrate, MES or N-oxides of these compounds, where

Ar1, Ar2and Ar3independently represent a five - or six-membered aryl, heteroaryl the second or cycloalkyl ring;

m is 0, 1, 2 or 3;

n and p independently are 0, 1, 2, 3 or 4;

r and t are independently 0, 1 or 2;

Y and Z are independently selected from the group including CR6R7, C=O, C=S, C=NOR6, O, NR6and S(O)b;

R1selected from the group comprising the SO2, C=O, C=S and C=NOR4;

X can be selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, -OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

X is preferably selected from the group comprising hydrogen, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, S(O)bR6, CONR6R7, -CO2R6, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6).

it is jdy R 1' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R2' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6) and P(O)(OR5)(OR6);

each R3/sup> ' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

or, alternatively, X and/or at least one of R1' together with the atoms to which they are linked, form an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cycloheptadiene or substituted cycloheptatriene ring, such ring may be fused with another aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cyclogeranyl or substituted cyclogeranyl ring;

R4-R8independently represent hydrogen, alkyl, substituted alkyl, aryl, Sames the config aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic or, alternatively, R5and R6, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring;

b is 0, 1 or 2.

According to another aspect, the invention proposed compounds having the structural formula (III)below:

or a salt, hydrate, MES or N-oxides of these compounds, where

Ar1, Ar2and Ar3independently represent a five - or six-membered aryl, heteroaryl or cycloalkyl ring, while Ar2and Ar3may be missing;

m is 0, 1, 2 or 3;

n and p independently are 0, 1, 2, 3 or 4;

each R1' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R2' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R3' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7/sup> R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

R5-R8independently represent hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic or, alternatively, R5and R6, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring;

b is 0, 1 or 2.

According to another aspect, the invention proposed a compound having the structure shown below:

or a salt, hydrate, MES or N-oxide of the specified connection.

According to another aspect, the invention proposed compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, salt is at or N-oxides of these compounds.

According to a related aspect, the proposed compound of structural formula (IV):

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar4and Ar5independently represent a five - or six-membered aryl or heteroaryl ring;

W is selected from the group including CR6R7, C=O, C=S; C=NOR6O, NR6, S, SO, SO2and (CH2)n;

n is 0, 1, 2 or 3;

G is selected from the group including CR6R7, C=O, C=S, C=NOR6and S(O)b;

R20selected from the group comprising hydrogen, arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R21selected from the group comprising arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R6and R7independently selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic, or, alternatively, R6and R7together with the atoms to which they are linked, form a loop which heteroalkyl or substituted cycloheptatriene ring; and

b is 0, 1 or 2.

According to yet another related aspect, the proposed compound of structural formula (V):

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar4and Ar5independently represent a five - or six-membered aryl or heteroaryl ring;

n is 0, 1, 2 or 3;

R21selected from the group comprising arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R35selected from the group comprising hydrogen, alkyl and substituted alkyl.

According to an additional variant of implementation, the proposed invention the compound of structural formula (VI)

or a salt, hydrate, MES or N-oxide of the compounds, where

R30selected from the group comprising arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R35selected from the group comprising hydrogen, alkyl and substituted alkyl.

According to an additional variant of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, with ivat or N-oxides of these compounds,

where each R independently represents Cl, MeO, CN, EtO, OH, Me, -SO2Me, F or H, and

n is 0, 1, 2, 3, or 4.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds,

where each R independently represents a MeO or OH, and

n is 0, 1, 2, 3, or 4.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds,

where R represents H, Me, Et, OCOMe, CH2OH, OMe or Ph.

According to the following variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

or Sol is, hydrate, MES or N-oxides of these compounds.

According to the following variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds.

According to one aspect, the present invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Ar6and Ar7that may be the same or different and independently from each other, represent a five - or six-membered aryl group or a five - or six-membered heteroaryl group;

Alk represents an alkyl group, optionally containing a heteroatom;

R36and R37that may be the same or different independently of one another, represent H, alkyl, or R36and R37together with the atoms to which they are attached, form an optionally substituted five - or six-membered heterocycle; and

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted shall heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated.

According to one aspect, the compounds according to the invention contain the five-membered heterocycle. According to one implementation variant, the five-membered heterocycle is an as or a substituted or unsubstituted cyclic urea.

According to one implementation variant, as is a as the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Ar6and Ar7that may be the same or different and independently from each other, represent a five - or six-membered aryl group or a five - or six-membered heteroaryl group;

Alk represents an alkyl group, optionally containing a heteroatom;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, C is displaced or the unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated; and

R39and R40that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated, or R39and R40together with the carbon atom to which they are attached, form a C=O group, or a substituted or unsubstituted alkenylphenol group.

According to another aspect, the compounds according to the invention contain the five-membered heterocycle, which is orasol. According to one implementation variant, orasol is orasol formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Ar6and Ar7that may be the same or different independently the Rog from each other, represent a five - or six-membered aryl group or a five - or six-membered heteroaryl group;

Alk represents an alkyl group, optionally containing a heteroatom;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated; and

R41represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated.

According to another aspect of the, compounds according to the invention contain six-membered heterocycle. According to one implementation variant, six-membered heterocycle represents a six-membered heterocycle of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated; and

R42, R43, R44, R45and R46that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, or R42and R43or R45and R46together with the carbon atoms to which they are both attached, form a C=O is the group.

According to another aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50represents H or substituted or unsubstituted alkyl;

R36and R37that may be the same or different independently of one another, represent H, alkyl, or R36and R37together with the atoms to which they are attached, form an optionally substituted five - or six-membered heterocycle; and

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl alkyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents the FDS is th N, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen.

According to another aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

T1represents C=O, and Q represents CR51R52or NR51,where R51and R52that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket substituted is whether the unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated, or R51and R52together with the carbon atom to which they are attached, form a C=O group, or a substituted or unsubstituted alkenylphenol group;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50represents H or substituted or unsubstituted alkyl;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or asamese the hydrated arylalkyl or halogen.

According to another implementation variant, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds.

According to another aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

T2represents the C=S, C=O or S(O)2;

R53represents a substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl;

M1represents N or CR54where R54represents H or substituted or unsubstituted alkyl;

M2before the hat is N or CR 55where R55represents H or substituted or unsubstituted alkyl;

R56represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R57represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen;

while this method involves reacting a compound of the formula:

where R56, R57and Alk are defined above, and J is a leaving group;

with the compound of the formula:

where M1and M2defined above, to obtain compounds of formula

,

contains NO2group;

the restoration of NO2group to obtain compounds containing NH2group; and

the interaction of compounds containing NH2the group, with the compound of the formula

where J2represents a leaving group, and T2and R53defined above.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or Sol is, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

R51and R52that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated, or R51and R52together with the carbon atom to which they are attached, form a substituted or unsubstituted alkenylphenol group;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50represents H or substituted or unsubstituted alkyl;

R38represents H, substituted or unsubstituted alkyl, substituted or asamese the hydrated cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen;

while this method involves the heating of the compounds of formula:

where R47, R48, Alk, M1and M2defined above;

for turning-CON3group-N=C=O group, and the subsequent interaction with the compound of the formula:

where J3represents a leaving group, and R38, R51and R52defined above.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or what if, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

R52represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50represents H or substituted or unsubstituted alkyl;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or not alseny arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen;

while this method involves the heating of the compounds of formula:

where R47, R48, Alk, M1and M2defined above;

for turning-CON3group-N=C=O group, and the subsequent interaction with a hydrazine of the formula:

where R38defined above.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar1, Ar2and Ar3independently represent a five - or six-membered aryl, heteroaryl or cycloalkyl ring, while Ar2and Ar3may be missing;

m is 0, 1, 2 or 3;

n and p independently is any 0, 1, 2, 3 or 4;

each R1' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R2' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR 6) and P(O)(R5)(OR6);

each R3' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

R5-R8independently represent hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic or, alternatively, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring;

b is 0, 1 or 2;

while this method involves reacting a compound of the formula:

where J represents the departure of the next group;

with the compound of the formula:

with the receipt of the product; and

the interaction of the specified product with the compound of the formula:

where J is a leaving group.

According to another aspect of the invention it is proposed to apply the substances identified using the assays described in the present description, as agents, or modulators of taste in the compositions in order to inhibit or block the bitter taste caused by substances that specifically activate specified in this description of the taste buds. A preferred object of the invention is the use of a substance that inhibits the activation of at least one of the above receptors T2R person to block the bitter taste of the substances present in coffee and food, beverages and medicines with the addition of coffee.

Another object of the invention is the use of the substances according to the present invention as inhibitors of the bitter taste of a wide range of actions in order to inhibit or block the bitter taste caused by substances that specifically activate the taste buds hT2R8, ligands that activate multiple taste buds, bitter substances with unknown specificity of the receptors, or compositions containing unknown bitter compounds, or the many bitter compounds. According to one variant of implementation, the substances according to the present invention is used for inhibiting the activation of at least one of the above receptors T2R person and, thus, block the bitter taste of the substances present in coffee and food, beverages and medicines with coffee flavor. Another object of the invention is to proof that the substances identified in accordance with the present invention, modulate, preferably inhibit or block the bitter taste caused, for example, coffee and foods, drinks and medicines with coffee flavor, with taste samples of human or animal, preferably using the taste of human samples.

Another object of the invention is the use of the substances identified using the assays described in the present text, as agents, or modulators of taste in the compositions in order to inhibit or block the bitter taste caused by substances that specifically activate specified in this description of the taste buds. A preferred object of the invention is the use of a substance that inhibits the activation of at least one specified the above receptors T2R person to block the bitter taste substances, present in coffee and food, beverages and medicines with the addition of coffee.

In the most preferred embodiment, the Substance and its analogues are used as inhibitors of the bitter taste of a wide spectrum of action for the inhibition or blocking of the bitter taste caused by substances that specifically activate the taste buds hT2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 64, 65, 71 and/or hT2R5, 9, 13, 54, 67 and 75, ligands that activate multiple taste buds, bitter substances with unknown specificity to receptors, or compositions containing unknown bitter compounds, or numerous bitter compounds. Given the antagonistic properties of the Substance, it must, in essence, to soften the bitter taste of the most bitter substances and containing such substances compositions. A preferred object of the invention is the use of a substance that inhibits the activation of at least one of the above receptors T2R person, such as a Substance, or its analog, in order to block the bitter taste of the substances present in coffee and food, beverages and medicines with the addition of coffee.

DETAILED DESCRIPTION of FIGURES

FIGURE 1 refers to experiments in which he used a partially purified bitter faction coffee for screening 25 T2R is the ne temporarily in transfected HEK cells, as described in previous patent applications included in the present description by reference. As shown in figure 1, the fraction coffee activated cells HEK293, temporarily transfetsirovannyh hT2R8 and hT2R14 in the analysis, using the methodology of the calcium image. To reduce the level of fluorescence fraction coffee, which could interfere with the analysis used a blue dye FD&C.

Figure 2 presents curve ΔF/F from the logarithm of the fraction of coffee, showing the dependence of the response hT2R8 dose fractions with a bitter taste, obtained from coffee. The analysis was performed using cell lines stably expressing hT2R8 and hT2R14, and automatic detector fluorescence FLIPR.

3 shows the curve of inhibition of the activity of hT2R8 in percent depending on the logarithm of the concentration of the substance showing the dependence of the inhibition dose-for Substances A and B in cell lines stably expressing hT2R8.

4 shows the curve of dependence of the inhibition activity hT2R14 a percentage of the logarithm of the concentration of Substances, demonstrating the dependence of the inhibition dose-for substances in cell lines stably expressing hT2R8.

FIGURE 5 shows the inhibitory activity of Substances for various (2) taste buds of the person.

Figure 6 presents the curve based cocktail recipes. the activity from the logarithm of the concentration of saccharin. Figure 6 shows the ratio of dose-response; and the influence of saccharin on the activity of receptors in transfected cells expressing variants hT2R43, hT2R44 and hT2R8. Answer hT2R8 on saccharin weaker in the in vitro than in alleles "tasters" hT2R43-W35 and hT2R44-W35, but it is stronger than the alleles of sadegustacio" hT2R43-S35 and hT2R44-R35.

DETAILED description of the INVENTION

In accordance with the present invention, the compounds according to the present invention can be used to mitigate or reduce the bitter taste of the compositions, for example, compositions for oral administration. In the present description, the term "composition for oral administration includes any substance intended for oral use, either individually or together with another substance. The composition for oral administration includes both "food or drink"and "food products". The term "food or drink" is understood as any food product intended for human or animal consumption, including solids, semi-solid substance or liquid (e.g., drinks). The term "non-food" or "food composition" includes supplements, nutraceuticals, functional foods (e.g., any fresh or processed food claimed as possessing properties, contributing the mi health promotion and/or properties to prevent diseases beyond basic nutritional function of supplying nutrients), pharmaceutical and over-the-counter medicines, products for care of the oral cavity, such as cleaning teeth and liquid mouthwash, cosmetic products such as lip balms, and other personal hygiene products.

The composition for oral administration also includes pharmaceutical, medicinal or food composition or, alternatively, the composition, for example, pharmaceutical or pharmaceutical composition or a food or beverage or composition.

Compounds according to the present invention can also be offered separately or in combination with any known or later open composition for ingestion. For example, the composition for oral administration can be a food or a food composition. The term "food composition" is understood any song that can be consumed as food by humans or animals, including solid, gel, paste, foam-like substance, semi-solid substances, liquids or mixtures of these substances. The term "non-song" understand any song that is intended for consumption or use by humans or animals not for food, including solid, gel, paste, foam-like substance, semi-solid substances, liquids or mixtures of these substances. Non-the song includes, without limitation, a pharmaceutical composition, which relates to food compositions intended for use by humans or animals for therapeutic purposes. The term "animal" includes any animal except man, such as farm animals and Pets.

In one implementation options of the compounds according to the present invention can be added to a food composition or food product, such as supplements, nutraceuticals, functional foods (e.g., any fresh or processed food claimed as possessing properties that promote health and/or properties to prevent diseases beyond basic nutritional function of supplying nutrients), pharmaceutical and over-the-counter medicines, products for care of the oral cavity, such as cleaning teeth and liquid mouthwash, cosmetic products such as lip balms, and other personal hygiene products.

In General, over-the-counter (OTC) product and a product for oral hygiene, as a rule, refer to the product for home and/or personal use, which can be sold without a prescription and/or without a visit to the doctor. Examples without prescription productdownload, without limitation, vitamins and dietary supplements; local analgesics and/or anesthetic; cough, colds and allergies; antihistamines and/or allergies; and combinations of these substances. Vitamins and dietary supplements include, without limitation, vitamins, food supplements, tonic beverages/bottled nutritive drinks, vitamins for children, food additives, and any other products that provide nutrients or related thereto, and combinations of these products. Local anaesthetics and/or anesthetic include any creams/ointments/gels for local application that is used to reduce the surface or deeply buried pain, for example, muscle pain; gel for pain relief during teething, patches analgesic component; and combinations of these substances. Cough, cold and Allergy include, without limitation, anti-inflammatory remedies, cough remedies, fringillinae medicines, medical confectionery, antihistamines and cough, colds and allergies for children; and combined products. Antihistamines and/or allergies include, without limitation, any system medicines for hay fever, nasal allergies, bites us comah. Examples of tools for oral hygiene include, without limitation, the cleaning strips for the mouth, toothpaste, toothbrushes, elixirs for oral/dental potions, care of dentures, mouth fresheners, bleach for teeth for use in the home, and floss.

In another embodiment, the implementation of the compounds according to the present invention can be added to foods and drinks or compositions. Examples of foods and drinks or compositions include, without limitation, coatings, glazes and glazing for food, and any substance included in the soups category dry technologically processed food, beverage category, the category of ready meals, category canned or tinned food, a category technologically frozen processed products category chilled technologically processed foods, snacks category, category, bakery products, category confectionery category dairy products, ice cream category, the category of substitutes food category pasta and noodles category and sauces, dressings, seasonings, category baby food and or category or pastes.

In General, the category of soups applies to canned/konservirovannoye, dry, fast food, chilled, the W-processed and frozen soup. For purposes of this definition, "soup (soups)" means a food made from meat, poultry, fish, vegetables, cereals, fruits and other ingredients, cooked in a liquid, which may contain visible pieces of some or all of the above ingredients. It can be transparent (broth) or thick (as thick stew), homogeneous, puree or contain pieces, ready-to-use, poluspuschennymi or condensed and can be served hot or cold as a first course or a main dish or as a smart snack (drink as a drink). The soup can be used as an ingredient for cooking other food components and can vary from soup (consommé) to sauces (butter or cheese soup).

The term "category dehydrated foods cooking" generally means (i) auxiliary products, such as powders, granules, pastes, concentrated liquid products, including concentrated broth, broth and polonophobia products in extruded cubes, tablets or powder or granulated form, which are sold separately in the form of a finished product or an ingredient in the product, sauces and mixtures prescription (regardless of technology); (ii) soluble foods, such as dry the UPA and soups freeze-drying, including dry soup mixes, dry instant soups, dry soup - semi-dry or not frozen compositions ready-to-eat foods, meals and snacks per serving, including pasta, potato and rice dishes; and (iii) products for decorating, such as seasonings, marinades, dressings for salads, toppings for salads, sauces for makanya, bread bread crumbs, whipped protein mix for breading, UHT pastes, sauces barbecue, liquid prescription mixes, concentrates, sauces or mixture of sauces, including salad, sold as a finished product or an ingredient in the product, dry or liquid, or frozen.

Category of drinks usually means drinks, mixes and concentrates beverages, including, without limitation, carbonated and non-carbonated drinks, alcoholic and non-alcoholic drinks, ready-to-drink beverages, concentrated liquid preparations for making beverages, such as soda water, and dry powder mixture precursor drinks. Drink category also includes alcoholic beverages, soft drinks, sports drinks, isotonic drinks and hot drinks. Alcoholic beverages include, without limitation, beer, cider/Perry, flavored alcoholic beverages (FABS), wine and spirits. Bezalkogolnoye include, without limitation, carbonated drinks, such as colas and fizzy drinks are not based on Cola, fruit juices, such as juices, nectars, juice drinks and drinks with the fruit taste; bottled water, which includes sparkling water, spring water and purified/dining room water, functional beverages which can be carbonated or non-carbonated, and include energy drinks or elixirs; concentrates, such as liquid and powder concentrates in ready-to-use dosage. Hot drinks include, without limitation, coffee, such as fresh (for example, welded), instant, combo coffee, liquid, ready-to-use, instant and dry coffee drinks, mixes and concentrates of coffee drinks, syrups, in pure form, in part or in powdered form; example "powder form" is a product containing coffee, sweetener and Dodge, all in powder form); tea, such as black, green, white, Oolong and flavored tea and other hot drinks, including flavored, malt or vegetable powders, granules, cubes, or pellets, mixed with milk or water.

Category snacks in General refers to any food product, which may be a light informal meal, including, without limitation, the smooth and savoury snacks and bars. Examples of snacks include, without limitation, fruit snacks, chips/tasted, extruded snacks, tortilla/corn chips, popcorn, pretzels, nuts, other sweet and savoury snacks. Examples of bars include, without limitation, the bars of granola/muesli bars for Breakfast, energy bars, fruit bars and other bars.

Category bakery products in General refers to any food product, the method of preparation which includes exposure to excessive heat or sunlight. Examples of bakery products include, without limitation, bread, buns, biscuits, muffins, a dish of cereal products, bread, confectionery, pastries, waffles, tortillas, biscuits, pies, donuts, tarts, quiches and a variety of fillings, cake, any baked products and any combination of these products.

Category ice cream in General relates to a frozen dessert containing cream and sugar, and flavoring. Examples include ice cream, without limitation, impulse ice cream; ice cream for consumption at home; frozen yogurt and homemade ice cream; soy, oatmeal, legumes (e.g., from red beans and mung beans) and rice ice cream.

Category confectionery generally refers to food PR is the product, sweet taste. Examples of confectionery products include, without limitation, candy, jelly, chocolate confectionery, confectionery sugar, chewing gum, etc. and any combination of products.

Category of meal replacements in General refers to any food product, intended for replacement of standard dishes, in particular for people who care about health or external form. Examples of meal replacements include, without limitation, slimming products and health products.

The ready meals category in General refers to any food product that may serve as food without the long cooking or processing. Ready meals include foods that contain added by the manufacturer prescription "on", resulting in a high degree of readiness, completeness and convenience. Examples of ready-made dishes include, without limitation canned/canned, frozen, chilled ready meals; dining mixture; frozen pizza; chilled pizzas; prepared salads.

Category pasta and noodles includes any pasta/noodles, including, without limitation, canned pasta, dried and chilled/fresh pasta; and a simple, instant, chilled, frozen noodles and noodles diner.

ategory canned/canned food includes, without limitation, canned/preserved meat and meat products, fish/seafood, vegetables, tomatoes, beans, fruit, ready meals, soup, pasta, and other canned/preserved food.

Category frozen food, processed, includes, without limitation, technologically frozen processed red meat, technologically processed poultry meat, technologically processed fish/seafood, technologically processed vegetables, meat substitutes, technologically processed potatoes, bakery products, desserts, ready meals, pizza, soup, noodles and other frozen food products.

Category dry food, processed, includes, without limitation, rice, dessert mixes, dried ready meals, dry soup, instant soup, dried pasta, plain noodles and instant noodles.

Category refrigerated food products, processed includes, without limitation, chilled technologically processed meat, technologically processed fish/seafood, kits for lunch, fresh-cut fruit, ready meals, pizza, salads, soup, fresh pasta, and noodles.

Category of sauces, dressings and condiments on the chaet, without limitation, tomato pastes and purees, bouillon/stock cubes, herbs and spices, monosodium glutamate (MSG), table sauces, soy sauces, sauces for pasta, wet/cooking sauces, dry sauces/powder mixes, ketchup, mayonnaise, mustard, dressings for salads, dressings with vinegar, sauces for makanya, pickles and other sauces, dressings and condiments.

Category baby food includes, without limitation, dairy or soy mixture; and cooked, dried and other baby food.

Category pastes include, without limitation, jams and preserves, honey, chocolate spread, peanut paste and yeast paste.

Category dairy products in General relates to a food product made from the milk of the mammal. Examples of dairy products include, without limitation, drinking dairy products, cheese, yoghurt and sour milk drinks and other dairy products.

Additional examples of food compositions, in particular food and drinks, or compounds as follows. Typical food compositions include one or more confectionery, chocolate confectionery, tiles, products intended for consumption on the go, chocolates with soft or hard stuffing in bags, assorted in boxes, standard assorted boxed chocolates with a backlog of the Oh ends "highly exacting twist wrapping applications", seasonal chocolate, chocolate with toys, alfajores (Argentine biscuits), other chocolate confectionery, mints, standard mints, strong mint candy, rock sugar, caramel, candy, chewing gum, jelly candies and chewing candies, toffees, caramels and nougat, medical confectionery, lollipops, licorice candy and other confectionery sugar, chewing gum, chewing gum with sugar, sugar-free gum, functional gum, inflatable chewing gum, bread, packaged/factory bread, unpackaged/homemade bread, flour pastries, cakes, packaged/factory cakes, unpackaged/home-made cakes, biscuits, chocolate-glazed cookies, layered biscuits, biscuits with fillings, savoury biscuits and crackers, bread substitutes, a mixture of grains for Breakfast, ready-to-eat cereal mixture, a mixture of grains for Breakfast for the whole family, flakes, muesli, other cereal mixture, a mixture of grains for Breakfast for children, hot cereal mixes, ice cream, impulse ice cream, dairy ice cream per portion ice cream on the basis of water per portion dairy ice cream Multipack, ice cream the basis of water Multipack, ice cream for consumption at home dairy ice cream to eat the house is, desserts ice cream, wholesale ice cream, dairy ice cream for consumption of home-based water, frozen yogurt, homemade ice cream, dairy products, milk, fresh/pasteurised milk, full fat fresh/pasteurised milk, semi-skimmed fresh/pasteurised milk, milk with a long shelf life/milk VIPs, full-fat milk with a long shelf life/milk VIPs, semi-skimmed milk with a long shelf life/milk VIPs, skim milk with a long shelf life/milk VIPs, goat milk, condensed/condensed sterilized milk, easy condensed milk/condensed sterilized milk, flavoured, functional and other condensed milk, flavored milk drinks, flavored milk drinks milk-based, flavored milk drinks with fruit juice, soy milk, sour milk drinks, fermented dairy drinks, clarifiers coffee (e.g., dairy and non-dairy cream or clarifiers for coffee drinks), milk powder, flavoured powder milk drinks, cream, cheese, processed cheese, spreads, processed cheese, nepostoyanniy melted cheese, napravlenii cheese pasty napravlenii cheese, hard cheese, packaged hard cheese, unpackaged hard cheese, yoghurt, p is hostoi/natural yoghurt, flavored yoghurt, fruit yoghurt, probiotic yoghurt, drinking yoghurt, regular drinking yoghurt, probiotic drinking yoghurt, chilled desserts and UHT desserts, dairy desserts, soy desserts, chilled snacks, soft cheese and curd low-fat, simple soft cheese and curd low-fat, flavored soft cheese and curd low-fat, salty soft cheese and curd low-fat, sweet and salty snacks, fruit snacks, chips/tasted, extruded snacks, tortilla/corn chips, popcorn, pretzels, nuts, other sweet and savoury snacks, snack bars, bars of granola, bars for Breakfast, energy bars, fruit bars, other snack bars, meal replacements, slimming products, health drinks, ready meals, canned ready meals, frozen ready meals, dried ready meals, chilled ready meals, dinner mixes, frozen pizza, chilled pizza, soup, canned soup, dry soup, instant soup, chilled soup, hot soup, frozen soup, pasta, canned pasta, dried pasta, chilled/fresh pasta, noodles, plain noodles, noodles quick cooking instant noodles in cups/bowls, noodles fast is prigotovleniya in packages, chilled noodles, eat noodles, canned goods, meat and meat products, canned fish/seafood, canned vegetables, canned tomatoes, canned beans, canned fruit, canned ready meals, canned soup, canned pasta, other canned foods, frozen foods, frozen technologically processed red meat, frozen technologically processed poultry, frozen technologically processed fish/seafood frozen technologically processed vegetables, frozen meat substitutes, frozen potatoes, baked potato chips, other baked potato products, nezamechennye frozen potatoes, frozen bakery, frozen desserts, frozen ready meals, frozen pizza, frozen soup, frozen noodles, other frozen foods, dehydrated products, dessert mixes, dried ready meals, dry soup, instant soup, dried pasta, plain noodles, instant noodles, instant noodles in cups/bowls, noodle packages, chilled food, chilled technologically processed meats, chilled technologically processed fish/seafood chilled technologically processed fish, chilled curd fish, chilled smoked fish, chilled set for lunch, chilled ready meals, chilled pizza, chilled soup, chilled/fresh pasta, chilled noodles, oils and fats, olive oil, vegetable oil and seed oil, cooking fats, butter, margarine, spreads, oils and fats, functional pasty oils and fats, sauces, dressings and condiments, tomato pastes and purees, bouillon/stock cubes, stock cubes, gravy granules, liquid broths and bases, herbs and spices, fermented sauces, soy sauces, sauces for pasta, wet sauces, dry sauces/powder mixes, ketchup, mayonnaise, regular mayonnaise, mustard, dressings for salads, conventional dressings for salads, low-fat dressings for salads, dressings with vinegar, sauces for makanya, pickles, other sauces, dressings and condiments, baby food, infant formula, the standard for infant formula, milk formula for children over one year of age, milk formula for toddlers who, hypoallergenic infant formula, prepared baby foods, dehydrated foods baby food, other baby foods, pasta, jams and preserves, honey, chocolate spread, peanut paste and yeast paste. Typical food compositions also include pastries, bread is baked goods, ice cream, dairy products, sweet and savory snacks, snack bars, meal replacements, ready meals, soups, pastas, noodles, canned foods, frozen foods, dehydrated foods, chilled foods, oils and fats, baby food or pastes or mixtures of these products. Typical food compositions also include a mixture of grains for Breakfast, sweet drinks or solid or concentrated liquid compositions for making beverages. Typical food compositions also include flavored coffee products (e.g. ice cream with the taste of coffee).

As a rule, sufficient to mitigate or reduce the bitter taste associated with the composition, for example a composition for oral administration add to the composition, resulting in the mitigation or reduction of the bitter taste associated with the specified composition in comparison with compositions obtained without compounds according to the present invention, focusing on people or animals. Or, in the case of the test compounds based on the majority of the group, for example, eight tasters (people)by procedures well-known in this field.

The concentration of the compounds according to the present invention, effective to mitigate or reduce the bitter taste, include the aqueous composition, of course, will depend on many factors, including the specific type of food composition and various other components of the natural genetic variability and individual preferences and health status of different people trying composition, and the subjective effects of certain compounds on the taste of the dynamics of such compounds. In some embodiments of the concentration of the compounds according to the present invention, effective to mitigate or reduce the bitter taste associated with the composition, is from about 0,001 million parts (ppm) to about 100 million parts, for example, from about 0.1 million parts to about 100 million units, from about 1 million to about 25 million parts, from about 1 million to about 10 million units, from about 0.1 million parts to about 10 million units, from about 0.01 million parts to about 30 million units, from about 0.05 million parts to about 10 million units, from about 0.01 million parts to about 5 million units, from approximately 0.02 million parts to about 2 million parts, or from about 0.01 million parts to about 1 million parts.

In some embodiments of the present invention provides that a mixture of one or more compounds according to the present invention will be used to mitigate or reduce the bitter taste associated with the composition. The end of the acidity of one or more compounds may be the same or concentration of each connection may be different.

Before further description of the present invention proposed the following definitions.

The term "T2R family" includes polymorphic variants, alleles, mutants and homologues, which (1) have about 30-40% identity of amino acid sequences, in particular about 40, 50, 60, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identity of amino acid sequences with receptors T2R described below and in the claims Zuker (Id) (2001) and Adler (Id.) (2001)included in the present description by reference, on a plot of approximately 25 amino acids, optimally 50-100 amino acids; (2) specifically bind to antibodies, raised to immunogen containing the amino acid sequence selected from the group comprising T2R sequence, described below, and conservatively modified variants of these sequences; (3) specifically hybridize (with a size of at least 100, optionally at least about 500-1000 nucleotides) in stringent conditions of hybridization to a sequence selected from the group comprising DNA sequences T2R described below, and conservatively modified variants of these sequences; (4) contain a sequence of at least 40% identical to the amino acid sequence selected from the group comprising amino acid sequences T2R, description is installed below or (5) of amplificatory primers that specifically hybridize in stringent conditions of hybridization in the above sequence T2R.

In particular, these receptors T2R include taste GPCR receptors, referred to in the present description hT2R8 and hT2R14 containing nucleic acid sequences and amino acid sequences proposed in this application, and variants, alleles, mutants, orthologues and chimeras of these sequences, which are specifically associated with bitter ligands identified in the present description, and other structurally related compounds and bitter compounds.

While T2R genes have significant sequence divergence at the level of proteins and DNA, it was found that all T2R allocated to date, contain some consensus sequences in certain areas, which are identical or have 70-75% sequence identity with the consensus sequence T2R previously identified in Adler et al. (WO 01/77676 A1 (2001) and Zuker et al. WO 01/18050 A2, both of which are fully incorporated into the present description by reference.

Topologically, some dynamics of GPCR contain N-terminal domain, extracellular domain, "transmembrane domain", containing seven transmembrane what Blasta and corresponding cytoplasmic and extracellular loops, "cytoplasmic region and the C-terminal region (see, e.g., Hoon et al., Cell, 96:541-51 (1999); Buck & Axel, Cell 65:175-87 (1991)). These areas can be structurally identified using methods known to experts in the art, such as for sequencing, to identify the hydrophobic and hydrophilic domains (see, for example, Stryer, Biochemistry, (3rd ed. 1988); see also any of a number of Internet programs for sequencing, such as on the website dot.imgen.bcm.tmc.edu). These areas are suitable for the creation of chimeric proteins for in vitro according to the present invention, for example, analyses of the ligand binding. For example, chimeric T2R can be obtained by combining the extracellular region of one T2R and the transmembrane region of another T2R same or different types.

Therefore, the term "extracellular region" refers to the domains of the polypeptides T2R, which protrude from the cell membrane and on the outward side of the cell. Such areas would include the N-terminal domain, which is on the outward side of the cell and the extracellular loops of the transmembrane domain, which is facing to the outside of the cell, i.e. the extracellular loop between transmembrane regions 2 and 3, transmembrane regions 4 and 5 and transmembrane regions 6 and 7. "The N-terminal domain starts at the N-terminal and p is ontiretse to the area close to the beginning of the transmembrane region. These extracellular region suitable for analysis of the ligand binding in vitro, as soluble, and such solid phase. In addition, the transmembrane region, described below, may also be included in the binding of ligands or in combination with extracellular region, or separately, and, therefore, also suitable for analysis of the ligand binding in vitro.

In the present description, the term "cell expressing T2R includes recombinant cells that Express a sequence T2R man according to the present invention, as well as endogenous cells expressing T2R. Such cells are found in lingual system and the gastrointestinal tract and include cells in the oral cavity, such as taste buds located on the tongue and cells in the gastrointestinal tract and related organs, such as brush cells in the gastrointestinal tract, enteroendocrine cells, such as cells STC-1. These cells can also Express the G-protein, such as gustducin, transducin, Gα15or Gα16. Cells that Express a specific T2R, can be identified and selected by known methods, as for example, by separation of cells by FACS method and/or procedures for selection of cells using magnetic microneedle./p>

"Transmembrane domain", containing seven transmembrane regions"refers to the domain T2R polypeptide, which is located inside the cell membrane, and may also include appropriate cytoplasmic (intracellular and extracellular loops, also called transmembrane "areas". The seven transmembrane regions and the extracellular and cytoplasmic loops can be identified by standard methods described in the Kyte &Doolittle, J. Mol. Biol., 157:105-32 (1982)) or in Stryer, above.

The term "cytoplasmic domains" refers to the domain T2R proteins, which are converted to the internal side of the cell, for example, "C-terminal domain and the intracellular loops of the transmembrane domain, the intracellular loops between transmembrane regions 1 and 2, a transmembrane regions 3 and 4 and transmembrane regions 5 and 6. The term "C-terminal domain" refers to the area that extends from the end of the last transmembrane region to the C-terminal of the protein and which, as a rule, is located in the cytoplasm.

The term "7-transmembrane receptor" means a polypeptide belonging to the superfamily of transmembrane proteins that contain seven areas covering the cell membrane seven times (thus, the seven areas are called "transmembrane" or " domains "TM" TM I-TM VII). Family obon is positive and some sensory receptors belong to this superfamily. The polypeptides of the 7-transmembrane receptor have similar and characterized by primary, secondary and tertiary structures, discussed in more detail below.

The term "binding ligand" refers to sequences derived from dynamics or gustatory receptor, which essentially include the transmembrane domains II-VII (TM II-VII). The specified region can bind the ligand and, in particular, the connection, causing the taste.

The term "translocation domain of the cell membrane" or simply "translocation domain" means a domain of the polypeptide, which when aminoterminal encoding the polypeptide sequence may with high efficiency "to accompany (to be a chaperone)or translational hybrid ("Chimera") protein in the cell membrane. For example, there may be used a specific "translocation domain", originally obtained from aminoterminal polypeptide adoptinga receptor human 7-transmembrane receptor. Another translocation domain was obtained from a sequence of bovine rhodopsin and is also suitable to facilitate translocation. Derived from the rhodopsin sequence, particularly effective in the translocation of 7-transmembrane hybrid proteins in the cell membrane.

The term "functional equivalence" means the ability and the power domain in the translocation of new translated proteins in the cell membrane as efficiently as a typical translocation domain, such as domain derived from rhodopsin in similar conditions; the relative effectiveness can be measured (in quantitative terms) and compared, as indicated in the present description. The domains included in the scope of the present invention, can be determined by standard screening their effectiveness in the translocation of newly synthesized polypeptides in the cell membrane in the cell (mammalian, Xenopus, and so on) with the same efficiency that the translocation domain of length 20 amino acids of SEQ ID NO:1.

The term "functional effects" in the context of assays for testing compounds that modulates the transmission of taste is mediated by a family member T2R, includes the determination of any parameter that is indirectly or directly under the influence of the receptor, e.g., functional, physical and chemical effects. It includes ligand binding, changes in ion flux, membrane potential, current flow, transcription, binding to G-protein, phosphorylation or dephosphorylation GPCR, signaling, interactions, receptor-ligand, the concentration of the second messenger (e.g. camp, cGMP, IP3, or intracellular Ca2+), in vitro, in vivo and ex vivo and also includes other physiologic effects such as increase or decrease the release of neuromedia the ditch or hormones.

The term "determining the functional effect" refers to tests for the presence of a compound that increases or decreases a parameter that is indirectly or directly under the influence of the T2R family member, e.g., functional, physical and chemical effects. Such functional effects can be measured by any means known to experts in the art, for example, changes in spectroscopic characteristics (e.g., fluorescence, absorbance, refractive index), hydrodynamic (e.g., shape), chromatographic properties or solubility, fixation capacity, potentialcustomers dyes, whole cell flows, radioisotope outflow induced markers, T2R gene expression in oocytes; T2R expression of tissue culture cells; transcriptional activation of T2R genes; analysis of ligand binding; voltage, membrane potential and conductance; analyses of ion flow; changes in intracellular second messengers such as camp cGMP and Insectivora (IP3); changes in intracellular levels of calcium; the release of neurotransmitters, etc.

The term "inhibitors," "activators," and "modulators" of protein receptors T2R are used interchangeably to refer to inhibitory, activating, or modulating molecules, identifizierung the x using in vitro and in vivo for the transfer of taste, for example, ligands, agonists, antagonists, and their homologs and mimetics. Inhibitors are compounds that, e.g., contact, partially or totally block stimulation, decrease, prevent, delay activation, inactivate, desencibiliziruta or inhibit the transmission of taste, e.g., agonists. Activators are compounds that, e.g., contact, stimulate, increase, start, activate, facilitate, enhance activation, sensibiliser or activate the transmission of taste, e.g., agonists. Modulators include compounds that, e.g., alter the interaction of the receptor with extracellular proteins that are associated with activators or inhibitor (e.g., barin and other members of the family of hydrophobic media); G-proteins; kinases (e.g., homologues of rhodopsin kinase and kinase beta-adrenergic receptors are involved in deactivation and desensitization of the receptor); and arrestin that also inactivate and desencibiliziruta receptors. Modulators include genetically modified variants of the T2R family members, e.g., with altered activity, as well as naturally occurring and synthetic ligands, antagonists, agonists, small chemical molecules and the like

Such assays for inhibitors and activators include, for example, the R, the implementation of the expression of members of the T2R family in the cells or cell membranes, applying alleged modulating compounds in the presence or in the absence of compounds that modulate, e.g bitter compounds, and then determining the functional effects on the transmission of taste, as described above. Samples or assays that contain members of the T2R family, which are a potential activator, inhibitor, or modulator are compared to control samples without the inhibitor, activator, or modulator, resulting examine the degree of modulation. Reference samples (not exposed to modulators) are assigned a relative activity T2R equal to 100%. Inhibition T2R carried out when the T2R activity value relative to the control is about 80%, optionally 50% or 25-0%. Activation of T2R carried out when the T2R activity value relative to the control is 110%, optionally 150%, optionally 200-500%, or higher 1000-3000%.

In the present description, the terms "purified", "essentially purified" and "isolated" refer to the condition of absence other than compounds with which the connection according to the present invention, generally associated in the natural state. Preferably, the terms "purified", "essentially purified" and "isolated" means that the components of ice contains at least about 0.5%, 1%, 5%, 10% or 20%, and most preferably at least 50% or 75% of the mass, the mass of this sample. In one of the preferred variants of realization of these terms refer to the connection according to the present invention, containing at least 95% weight by weight of this sample. In the present description, when the terms "purified", "essentially purified" and "isolated" refers to nucleic acid or protein, nucleic acids or proteins, also indicate the state of purification or concentration different from that which occurs in nature in the body of a mammal, especially man. Any purification or concentration, more naturally occurring in the body of a mammal, especially human, including (1) clearance from other related structures or compounds or (2) communicate with the structures or compounds with which it is usually not connected in the body of a mammal, especially man, is included in the meaning of the term "isolated". Nucleic acid or protein or classes of nucleic acids or proteins mentioned in the present description, may be highlighted or otherwise associated with structures or compounds with which they usually are not associated in nature according to a series of methods and processes known to specialists in this field of technology.

In the present description, the term "vydeleny is" in relation to nucleic acid or polypeptide refers to a state of purification or concentration, different from that which occurs in nature in the body of a mammal, especially man. Any purification or concentration, more naturally occurring in the body, including (1) clearance from other naturally occurring associated structures or compounds or (2) communicate with the structures or compounds with which it is usually not connected in the body, in the present description is included in the meaning of the term "isolated". Nucleic acids or polypeptides described in the present description, may be highlighted or otherwise associated with structures or compounds with which they usually are not associated in nature according to a series of methods and processes known to specialists in this field of technology.

In the present description, the terms "amplificating" and "amplification" refers to the use of any suitable amplification method for receiving or detecting or recombinant expressed in the nature of nucleic acids, as described in more detail below. For example, according to the present invention methods and reagents (e.g., specific pairs of oligonucleotide primers for amplification (e.g., by polymerase chain reaction, PCR) expressed in nature (e.g., genomic or mRNA) or recombinant (e.g., cDNA) nucleic acids according to the present image the structure (for example, sequences according to the present invention, communicates with the connection, causing the taste) in vivo or in vitro.

The term "expression vector" refers to any system for recombinant expression for the expression of nucleic acid sequence according to the present invention in vitro or in vivo, constitutively or induced in any cell, including prokaryotic, yeast, fungal, plant cell, insect cell or mammal. The term includes linear or ring system for expression. The term includes systems for expression, which remain episomal form or integrate into the genome of the host cell. System for the expression may or may not be able or infect other programs, i.e., to exercise only a temporary expression in the cell. The term includes recombinant expression "cassette"containing only the minimum number of elements required for transcription of the recombinant nucleic acids.

The term "library" means a composition that is a mixture of different nucleic acid molecules or polypeptides, as for example, a library of recombinant received touch, in particular regions of the binding ligand taste buds obtained by amplification of nucleic acid with pairs of degenerate primers or the dedicated set of vectors, containing amplificatoare region binding ligand, or mixture of cells, each randomly transfusiona at least one vector encoding a taste receptor.

The term "nucleic acid" or "nucleic acid sequence" refers to an oligonucleotide, deoxyribonucleotide or ribonucleotide, either single-stranded or double-stranded form. The term includes nucleic acids, i.e. oligonucleotides, containing known analogues of natural nucleotides. The term also includes structures, such as nucleic acids, synthetic cores.

Unless otherwise stated, provided that the specific sequence of nucleic acids also include conservatively modified variants of the sequence (for example, substitution of degenerate codons) and complementary sequences, as well as a clearly defined sequence. In particular, substitution of degenerate codons can be carried out by receiving, for example, sequences in which the third position of one or more selected codons substituted with mixed-base and/or the remains of deoxyinosine (Batzer et al., Nucleic Acid Res., 19:5081 (1991); Ohtsuka et al., J. Biol. Chem., 260:2605-08 (1985); Rossolini et al., Mol. Cell. Probes, 8:91-98 (1994)). The term "nucleic acid" is used interchangeably with gene, cDNA,mRNA, the oligonucleotide and polynucleotide.

In the present description, the terms "polypeptide", "peptide" and "protein" are used interchangeably and refer to a polymer of amino acid residues. These terms of use to amino acid polymers in which one or more amino acid residues is an artificial chemical mimetic of a corresponding naturally occurring amino acids, as well as to naturally occurring amino acid polymers and not naturally occurring amino acid polymer.

In the present description, the term "alkyl", as well as other groups with the prefix "ALK"such as, for example, alkoxy, alkanoyl, alkenyl, quinil and the like, means carbon chains which may be linear or branched, or a combination of these circuits. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec - and tert-butyl, pentyl, hexyl, heptyl and the like, Preferred alkyl groups contain 1-4 carbon atoms. The term "alkenyl" and other similar terms include carbon chain containing at least one unsaturated bond carbon-carbon. The term "quinil" and other similar terms include carbon chain containing at least one triple bond of carbon-carbon.

The term "cycloalkyl" means carbocycle, not containing heteroatoms, and includes the em mono-, bi - and tricyclic saturated carbocycles, as well as the condensed ring system. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, decahydronaphthalene, adamantane, indanyl, indenyl, fluorenyl, 1,2,3,4-tetrahydronaphthalen etc.

The term "aryl" means aromatic Deputy, which is a single ring or multiple rings fused with each other. Typical aryl groups include, without limitation, phenyl, naftalina, antarctilyne, pyridinoline, personilnya, pyrimidinyl, triazinyl, tiffaniejoy, fornillo, pyrrolidino, oxazolidinyl, imidazolidinyl, triazolyl and tetrazolyl group. Aryl groups containing one or more heteroatoms (e.g., pyridinyl), often referred to as "heteroaryl groups". In the formation of multiple rings at least one of the members of the rings is aromatic. In some embodiments of at least one of the multiple rings contains a heteroatom, thus forming an aryl group containing a heteroatom. Aryl group containing a heteroatom include, without limitation, benzoxazolyl, benzimidazolyl, khinoksalinona, benzofuranyl and 1H-benzo[d][1,2,3]triazolyl group. Aryl group containing a heteroatom, that the same include, without limitation, 2,3-dihydrobenzo[b][1,4]dioxinlike and benzo[d][1,3]dioxolo group. Aryl group containing a heteroatom include aromatic rings, condensed with heterocyclic ring containing at least one heteroatom and at least one carbonyl group. Such groups include, without limitation, dioxotetrahydrofuran and dioxotetrahydrofuran group.

The term "Allakaket" means aryl group associated with alkoxygroup.

The term "arylamidase" means an aryl-C(O)NR-alkyl or aryl-NRC(O)-alkyl.

The term "arylalkylamines" means an aryl-alkyl-C(O)NR-alkyl or aryl-alkyl-NRC(O)-alkyl, where R is any suitable group, below.

The term "arylalkyl" refers to an aryl group linked to an alkyl group.

The term "halogen" or "halogen" refers to chlorine, bromine, fluorine or iodine.

The term "leaving group" refers to a functional group or atom which can be substituted with another functional group or another atom in a substitution reaction, such as reaction of nucleophilic substitution. As an example, a typical leaving groups include chlorine, bromine and iodine group; groups, sulfonic ester, such as mesilate, toilet, brasilit, nosrat and the like; and alloctype, such as acetoxy, triflora is ethoxy etc.

The term "halogenated" means an alkyl group containing one or more halogen atoms (for example, CF3).

The term "heteroalkyl" refers to an alkyl group containing a heteroatom such as N, O, P, B, S, or Si. The heteroatom may be connected to the rest heteroalkyl group means a saturated or unsaturated communication. Thus, alkyl containing as substituents group, such as heteroseksualci, substituted heteroseksualci, heteroaryl, substituted heteroaryl, alkoxy, aryloxy, boryl, phosphino, amino, silyl, thio or selenium, is included in the term "heteroalkyl". Examples of heteroalkyl include, without limitation, cyano, benzoyl, 2-pyridyl and 2-furyl.

The term "heteroaromatic" means a heteroaryl group that is attached to an alkyl group.

The term "heterocycle" means a monocyclic or polycyclic ring containing carbon atoms and hydrogen, optionally containing one or two multiple bonds; however, the atoms in the ring contains at least one heteroatom, in particular 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur. Patterns of heterocyclic rings include, without limitation, mono-, bi - and tricyclic compounds. Specific compounds are monocyclic or bicyclic. Typical heterocycles include cyclo is acheinu, morpholinyl, pyrrolidinyl, pyrrolidinyl, piperidinyl, piperazinil, hydantoinyl, valerolactam, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridine, tetrahydropyrimidines, tetrahydrothiophene, tetrahydrothiopyran, tetrazolyl and ursolic. The heterocyclic ring can be substituted or unsubstituted, preferably, the heterocycles are 5 - or 6-membered heterocycles, in particular, hydantoinyl and ursolic.

The term "heteroseksualci" refers to cycloalkyl group in which at least one of the carbon atoms in ring substituted on a heteroatom (such as O, S or N).

The term "geterotsiklicheskikh" means geterotsyklicescoe group is attached to an alkyl group.

The term "containing substituents, in particular, provides for and allows one or more substitutions accepted in the art. However, specialists in the art it is obvious that the deputies should be chosen in such a way as not to adversely impact on the useful characteristics of the connection, or not to interfere with its functions. Suitable substituents can include, for example, galactography, performanceline group, performancebuy, alkyl groups, alkeline group, alkyline group, a hydroxy-group, carbonyl group, mercaptopropyl, is Citigroup, alkoxygroup, aryl or heteroaryl group, aryloxy or heterokaryosis, kalkilya or heteroalkyl group, Alcoxy or heteroaromatic, amino groups, alkyl - and dialkylamino, carbamoyl group, alkylcarboxylic group, carboxyl group, alkoxycarbonyl group, alkylaminocarbonyl group, dialkylaminoalkyl group, arylcarbamoyl group, aryloxyalkyl group, alkylsulfonyl group, arylsulfonyl group, cycloalkyl group, ceanography, ancilliary C1-C6, aristocraty, nitro, geograpy, acyl group, boronate or boronlike groups, phosphate or postonline group, sulfanilimide group, sulfonylurea group, sulfinyl groups and combinations of these groups. In the case of containing substituents combinations, such as "containing substituents arylalkyl", or aryl or alkyl group may contain substituents, or aryl, and alkyl groups may contain one or more substituents. In addition, specialists in the art it is known that in some cases, suitable substituents may be combined to form one or more rings.

The compounds specified in the present description, contain one or more double bonds and thus the time can lead to CIS/TRANS-isomers, and other conformational isomers. The present invention includes all the possible isomers of this kind, as well as mixtures of such isomers.

The compounds specified in the present description, and in particular the above substituents can have one if several centers of asymmetry and thus can lead to diastereomers and optical isomers. In the present invention includes all possible diastereomers of this kind, as well as their racemic mixtures, their essentially pure allocated enantiomers, all possible geometric isomers, and acceptable salts of these isomers. In addition, also included a mixture of stereoisomers, as well as the specific selected stereoisomers. In the process of synthesis procedures used to obtain these connections or using procedures racemization and epimerization, well-known specialists in the field of technology products such procedures can be a mixture of stereoisomers.

In the present description, the terms "salt" and "pharmaceutically acceptable salts" refer to derivatives of these compounds, when the original connection modify by obtaining acidic or basic salts of this compound. Examples of pharmaceutically acceptable salts include, without limitation, salts of inorganic or organic acids of the basic group is, such as amines; alkali or organic salts of acidic groups such as carboxylic acid. Pharmaceutically acceptable salts include conventional non-toxic salts or the salts of Quaternary ammonium parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric, Hydrobromic, sulfuric, sulfamic, phosphoric and nitric; and salts derived from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, Panova, maleic, hydroxymaleimide, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluensulfonate, methanesulfonate, ethicality, oxalic and setinova etc.

Pharmaceutically acceptable salts according to the present invention can be synthesized from the parent compound that contains a basic or acidic group, standard chemical methods. Generally, such salts can be obtained by interaction of free acidic or basic forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or organic process is the or mixtures thereof; in General, preferred are anhydrous environment like ether, ethyl acetate, ethanol, isopropanol or acetonitrile. Lists of suitable salts are listed in theRemington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, the disclosure of which is incorporated into this description by reference.

The term "MES" means a compound or salt of the compounds, which further comprises a stoichiometric or non-stoichiometric amount of solvent, ecovalence associated forces of intermolecular interaction. When the solvent is a water, MES is a hydrate.

The term "prodrug" means a derivative of a compound that can be either hydrolyzed, oxidize, or otherwise react in biological conditions (in vitroorin vivowith the formation of the active compounds, in particular compounds according to the present invention. Examples of prodrugs include, without limitation, derivatives and metabolites of the compounds according to the present invention, which include biohydrology compounds such as biokerosene amides, biohydrology esters, biohydrology carbamates, biohydrology carbonates, biohydrology ureides and biokerosene analogues of phosphate. Specific prodrugs of compounds with carboxyl functional g is uppada constitute the lower alkylether carboxylic acid. Carboxylate esters are conveniently obtained by implementing the esterification of any of the carboxylic acid group in the molecule. Typically, the prodrug can be obtained by well known methods such as describedBurger''s Medicinal Chemistry and Drug Discovery6th ed. (Donald J. Abraham ed., 2001, Wiley) andDesign and Application of Prodrugs(H. Bundgaard ed., 1985, Harwood Academic Publishers Gmfh).

In the present description, unless otherwise specified, the terms "biohydrology amide", "biohydrology ether", "biohydrology carbamate", "biohydrology carbonate", "biohydrology of wreid", "biohydrology phosphate" means amide, ester, carbamate, carbonate, wreid or phosphate, respectively, compounds that either 1) does not interfere with the biological activity of the compound but can confer this connection is useful properties in vivo, such as uptake, duration of action, or onset of action; or 2) is biologically inactive but is converted in vivo to the biologically active connection. Examples biohydrology esters include, without limitation, lower alkylamine, alkoxylation, alkylarylpolyglykelether and kalinovye esters. Examples biohydrology amides include, without limitation, lower alkylamide, amides of alpha-amino acids, alkoxysilane and alkylaminocarbonyl. Examples biohydrology carbamates include, without limitation, lower alkylamines followed, with the holding deputies ethylendiamine, amino acids, hydroxyethylamine, heterocyclic and heteroaromatic amines and polyetheramines.

In the present description, the term "analogue of the specified connection" in the context of the compounds specified in the present description includes the diastereomers, hydrates, solvate, salt, prodrugs and N-oxides of the compounds.

"Translocation domain", "binding ligand" and the composition of the chimeric receptors, referred to in this description, also include "analogs," or "conservative variants" and "mimetics" ("peptidomimetics")with structures and activity that essentially corresponds to the typical sequences. Thus, in the present description the terms "conservative variant" or "analog"or "mimetic" refer to a polypeptide containing a modified amino acid sequence, such that the change (change) is not essentially change the structure and/or activity of the polypeptide (conservative version). These terms include conservatively modified variations of amino acid sequences, i.e. substitutions, additions of amino acids or deletions those residues that do not have critical values for the protein activity or substitution of amino acid residues having similar properties (e.g., acidic, basic, positively or negatively the charger, side buttons is installed, polar or non-polar etc), so that overrides even the key amino acids is not essentially change the structure and/or activity.

More specifically, the term "conservatively modified variants" applies to both amino acid sequences and sequences of nucleic acids. In respect of specific sequences of nucleic acids, the term "conservatively modified variants" means those nucleic acids that encode identical or essentially identical amino acid sequences, or in the case where the nucleic acid does not encode the amino acid sequence, refers to essentially identical sequences. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein.

For example, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be replaced by any of the described respective codons without changing the encoded polypeptide.

Such variations of nucleic acids are "silent variations", which are one type of conservatively modified variations. In the present description, each sequence of nucleic acid, Cody the respective polypeptide, also describes every possible silent variation of the nucleic acid. Specialist it is obvious that each codon in a nucleic acid (except AUG, which, as a rule, is the only codon for methionine, and TGG, which, as a rule, is the only codon for tryptophan) can be modified to obtain a functionally identical molecule. Therefore, each silent variation of a nucleic acid encoding a polypeptide is implicit in each described sequence.

Table a conservative substitution, which are functionally similar amino acids are well known in the art. For example, one of the typical principles select conservative substitutions includes (original residue followed by a typical substitution): ala/gly or ser; arg/lys; asn/gln or his; asp/glu; cys/ser; gin/asn; gly/asp; gly/ala or pro; his/asn or gln; ile/leu or val; leu/ile or val; lys/arg or gln or glu; met/leu or tyr or ile; phe/met or leu or tyr; ser/thr; thr/ser; trp/tyr; tyr/trp or phe; val/ile or leu. According to an alternative of the typical principle uses the following six groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), serine (S), Threonine (T); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (I);5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); (see also, e.g., Creighton, Proteins, W. H. Freeman and Company (1984); Schultz and Schimer, Principles of Protein Structure, Springer-Verlag (1979)). Specialist in the art it is obvious that videopreteen substitution are not the only possible one conservative substitution. For example, for some purposes can be considered to be all charged amino acid conservative substitutions for one another, positive or negative. In addition, individual substitutions, deletions or additions which alter, add, or delete a single amino acid or a small percentage of amino acids in the encoded sequence can also be viewed as a "conservatively modified variations".

The terms "mimetic" and "peptidomimetic" refer to a synthetic chemical compound which has essentially the same structural and/or functional characteristics of the polypeptides, for example, translocation domains, regions of the binding ligand or chimeric receptor according to the present invention. The mimetic can be either entirely composed of synthetic, non-natural analogues of amino acids, or may be a chimeric molecule of partly natural peptide amino acids and partly non-natural analogs of amino acids. Mimetic is the same to contain any number of conservative substitution of natural amino acids, provided that such substitution is also not fundamentally change the structure and/or activity of mimetica.

As in the case of polypeptides according to the present invention which are conservative variants, routine testing will determine whether a mimetic in the scope of the present invention, i.e., that its structure and/or function is not essentially changed. Composition of mimetic polypeptide can contain any combination of non-natural structural components, which generally are composed of three structural groups: a) group clutch residues other than the natural amide bonds (peptide bonds"); (b) non-natural residues instead of the naturally occurring amino acid residues; or C) residues which induce the mimicry of the secondary structure, i.e. induce or stabilize a secondary structure, for example, beta-turn, gamma-turn, beta-layer conformation of alpha-helix and the like Polypeptide may be characterized as a mimetic, when all or several residues are connected by chemical rather than natural peptide bonds. Individual coworkers peptide residues can be joined by peptide bonds, other chemical bonds or by conjugation, such as glutaraldehyde, N-hydroxysuccinimide esters, bifunctional maleimides, N,N'-dicyclohexylcarbodiimide (DCC or N,N'-diisopropylcarbodiimide (DIC). Linking groups that can be an alternative to the traditional amide linkages ("peptide bond") include, for example, kilometre (for example, --C(.=O)--CH2for --C(.=O)--NH--), iminomethylene (CH2NH), ethylene, olefin (Snavely swazis), ether (CH2O), thioether (CH2--S), tetrazole (CN4), thiazole, retreated, thioamide or ester (see, for example, Spatola, Chemistry and Biochemistry of Amino Acids, Peptides and Proteins, Vol. 7, 267-357, Marcell Dekker, Peptide Backbone Modifications, NY (1983)). The polypeptide may also be characterized as a mimetic, if it contains all or some non-natural residues instead of the naturally occurring amino acid residues; non-natural residues are well described in the scientific and patent literature.

"Label" or "contrast agent" is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, or chemical means. For example, suitable labels include32P, fluorescent dyes, electron-dense reagents, enzymes (e.g., commonly used in an ELISA), Biotin, digoxigenin or haptens and proteins which can be made detectable, e.g., by introducing a radioactive label to the peptide, or which can be used to detect antibodies specifically reactive with the peptide.

"Labeled probe nucleic acid or oligonucleotide" not only is em a probe, which is associated either covalently, through a linker or a chemical bond, or are not covalently bound through ionic, van der Waals, electrostatic, or hydrogen bonds to a label such that the presence of the probe may be detected by detecting the presence of label associated with the probe.

In the present description, the term "probe nucleic acid or oligonucleotide" is defined as a nucleic acid capable of contacting the target nucleic acid of complementary sequence through one or more types of chemical bonds, usually through complementary mating grounds, usually by formation of hydrogen bonds. In the present description, the probe may include natural (i.e. A, G, C or T) or modified bases (7-deazaguanosine, inosine and so on). In addition, the bases in the probe can be connected by a connection other than phosphodiester bond, provided that it does not interfere with hybridization. Thus, probes can be a peptide-nucleic acids in which the constituent bases are joined by peptide bonds rather than phosphodiesterase links. Specialist in the art it is obvious that the probes may be contacted with the target sequences lacking complete complementarity with the sequence of the probe, depending on with whom agosti conditions of hybridization. The probes can be directly outline as with isotopes, chromophores, phosphors, Chromogens, or indirectly in the state as in the case of Biotin, which streptavidin complex can be contacted later. By conducting analysis on the subject the presence or absence of the probe to detect the presence or absence of the selected sequence or subsequence.

The term "heterologous" when used in relation to the parts of the nucleic acid indicates that the nucleic acid contains two or more subsequences that nature is in a different relationship with each other. For example, nucleic acid, typically receive a recombinant method, and it contains two or more sequences from unrelated genes arranged to make a new functional nucleic acid, for example, a promoter from one source, and the coding region from another source. Similarly, the term "heterologous protein" means that the protein contains two or more sequences, which in nature is in a different relationship to each other (for example, a hybrid protein).

The term "promoter" is defined as a set of nucleic acids sequences which direct transcription of the nucleic acid. In the present description is a promoter includes necessary nucleic acid sequences near the site of transcription initiation, such as in the case of promoter polymerase type II, a TATA element. The promoter also optionally includes distal elements enhancer (amplifier) or repressor, which can be located at a distance of up to several thousand base pairs from the site of transcription initiation. "Constitutive promoter" is a promoter that is active under most environmental conditions environmental and experimental conditions. "Inducible promoter" is a promoter that is active in the regulation of environmental and experimental regulation. The term "functionally linked" refers to functional relationships between the nucleic acid sequence controlling the expression (such as a promoter or set of centers (sites) binding of transcription factors), and the second sequence of nucleic acid, when controlling the expression sequence directs transcription of the nucleic acid corresponding to the second sequence.

In the present description, the term "recombinant" refers to polynucleotide, synthesized or otherwise affected in vitro (e.g., "recombinant polynucleotide"), to methods of using recombinant polynucleotides for obtaining gene products in cells or other biological systems, or to a polypeptide (the recombinant protein"), encoded by recombinant polynucleotide. The term "recombinant means" also includes ligation of nucleic acids containing various coding region or domains or sequences of promoters from different sources in the expression cassette or vector for expression of, e.g., inducible or constitutive expression of a hybrid protein containing the translocation domain according to the present invention and the sequence of the nucleic acid amplified using the primer according to the present invention.

The term "selectively (or specific) hybridizes" refers to the binding, the formation of duplex or hybridizing of a molecule only to a particular nucleotide sequence in the hard hybridization conditions when that sequence is present in a complex mixture (e.g., total cellular or library DNA or RNA).

The term "stringent hybridization conditions" refers to conditions under which a probe will be gibridizatsiya in the target sequence, typically in a complex mixture of nucleic acid, but not in the other sequences. Hard conditions depend on the sequence and will be different in different circumstances. Longer sequences, in particular, hybridize at higher temperatures. P is the fractional guide to the hybridization of nucleic acids described in Tijssen, Techniques in Biochemistry and Molecular Biology--Hybridization with Nucleic Probes, "Overview of principles of hybridization and the strategy of nucleic acid assays" (1993). In General, stringent conditions are selected so as to be about 5-10°C. below the melting temperature (Tm) for the specific sequence at a defined ionic strength pH. TA represents the temperature (under defined ionic strength, pH, and concentration of nucleic acid)at which 50% of the probes complementary to the target hybridize to the target sequence at equilibrium (as the target sequences are present in excess, at Tm, 50% of the probes are in equilibrium). Stringent conditions are conditions where the salt concentration is less than about 1.0 M concentration of sodium ion, typically about 0.01 to 1.0 in M concentration of sodium ion (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes (e.g., 10 to 50 nucleotides) and at least about 60°C for long probes (e.g., more than 50 nucleotides). Stringent conditions can also be obtained by the addition of destabilizing agents such as formamide. For selective or specific hybridization, a positive signal represents at least twice, not necessarily ten times background hybridization. Typical stringent hybridization conditions can be as follows: 50% of formamide, 5×SSC, and 1% of the LTO, implementation incubation at 42°C, or 5×SSC, 1% LTOs, implementation incubation at 65°C with washing in 0.2×SSC and 0.1% LTOs at 65°C. Such stages of hybridization and washing can be performed, for example, within 1, 2, 5, 10, 15, 30, 60 or more minutes.

Nucleic acids that do not hybridize to each other under stringent conditions, however, are essentially related, if the polypeptides which they encode, are essentially related. This occurs, for example, when a copy of a nucleic acid is produced using the maximum degeneracy of codon permitted by the genetic code. In such cases, nucleic acid, typically hybridize in moderately stringent conditions of hybridization. Typical "moderately stringent hybridization conditions" include a hybridization in a buffer of 40% formamide, 1 M NaCl, 1% LTOs at 37°C and washing in 1×LTOs at 45°C. Such stages of hybridization and washing can be performed, for example, within 1, 2, 5, 10, 15, 30, 60 or more minutes. Positive hybridization is at least twice background. Any specialist it is obvious that the alternative conditions of hybridization and washing can be used to obtain conditions similar stiffness.

The term "antibody" refers to a polypeptide that contains a wireframe plot of the immunoglobulin gene or its fragments is s, which specifically binds and recognizes an antigen. The recognized immunoglobulin genes include the Kappa, lambda, alpha, gamma, Delta, Epsilon and mu-genes constant region, as well as countless immunoglobulin gene variable regions. Light chains are classified into the Kappa or lambda. Heavy chains are classified into gamma, mu, alpha, Delta, or Epsilon, which in turn define classes of immunoglobulins, IgG, IgM, IgA, IgD and IgE, respectively.

The structural unit of a typical immunoglobulin (antibodies) contains a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each of which contains one "light" (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The N-terminal of each chain defines a variable region of about 100-110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) attributable to the said light and heavy chains, respectively.

"Chimeric antibody" is an antibody molecule in which (a) the constant region or part thereof altered, replaced or exchanged so that antigennegative centre (variable region) is linked to a constant region of a different or altered class, effector function and/or widely completely different molecules, which adds new properties to the chimeric antibody, e.g., enzyme, toxin, hormone, growth factor, drug, etc.; or (b) a variable region or part thereof altered, replaced or exchanged for a variable region having a different or altered specificity towards the antigen.

Antibody anti-T2R" is an antibody or antibody fragment which is specific binds to the polypeptide encoded T2R gene, cDNA, or a subsequence.

The term "immunological analysis" is an analysis using antibodies for specific binding to the antigen. Immunological analysis is characterized by the use of specific binding properties of a particular antibody to isolate, target, and/or to quantify the antigen.

The term "specific (or selectively) binds" to an antibody or specific (or selectively) immunoreactive with respect to" when used to a protein or peptide means the binding reaction, which determines the presence of the protein in a heterogeneous population of proteins and other biological forms. Thus, in the indicated conditions immunoassay specific antibodies bind to a particular protein at least twice and essentially does not bind in significant the considerable number of other proteins, present in the sample. For specific binding to an antibody under such conditions may require an antibody that is chosen on the basis of its specificity towards a particular protein.

For example, polyclonal antibodies, raised to the T2R family member of a specific species such as rat, mouse, or human can be selected to obtain only those polyclonal antibodies that are specific immunoreactive against the T2R polypeptide or immunogenic part, and not to other proteins with the exception of orthologues or polymorphic variants and alleles of the T2R polypeptide. This selection can be done by removing antibodies that give cross-react with molecules T2R other species or other molecules T2R. Can also be selected only those antibodies that recognize GPCR only the T2R family, and not other GPCR families. Can be used a number of formats immunological analysis to select antibodies specific immunoreactive against a specific protein. For example, solid phase immunological assays ELISA assays (ELISA) are usually used to select antibodies specific immunoreactive with respect to the protein (see, for example, Harlow &Lane, Antibodies, A Laboratory Manual, (1988) for a description of the formats and conditions immunoassay that can be used to determine the SPE is oficinas immunoreactivity). Typically a specific or selective reaction will constitute at least twice background signal or noise, and mostly more than 10-100 times the background signal or noise.

The term "selectively associated with" refers to the ability of the nucleic acid selectively gibridizatsiya" with another nucleic acid, as defined above, or to the ability of antibodies "selectively (or specific) to contact with the protein, as defined above.

The term "expression vector" refers to any system for recombinant expression for the expression of nucleic acid sequence according to the present invention in vitro or in vivo, constitutively or induced in any cell, including prokaryotic, yeast, fungal, plant cell, insect cell or mammal. The term includes linear or ring system for expression. The term includes systems for expression, which remain episomal form or integrate into the genome of the host cell. System for the expression may or may not be able or infect other programs, i.e., to exercise only a temporary expression in the cell. The term includes recombinant expression "cassette"containing only the minimum number of elements required for transcription of the recombinant nucleic acids.

the term "a host cell" means the cell, which contains an expression vector and promotes the replication or expression of the expression vector. Cell host can be a prokaryotic cell such as E. coli, or eukaryotic cells such as yeast cells, insect, amphibian or mammal, such as CHO, HeLa, HEK-293 and the like, for example, cultured cells, explants and cells in vivo.

Based on the foregoing, according to the present invention proposed assays to identify compounds that modulate, preferably block, specific activation of previously identified receptor bitter taste of the person by means of bitter compounds, for example, bitter compounds present in coffee and obtained from him the extracts, and structurally related and other bitter compounds. In particular, according to the present invention proposed tests based on the cells to perform identification of compounds that modulate (e.g. inhibit) the activation of hT2R8 and hT2R14. These connections will modulate bitter taste associated with these taste receptors in humans. This will be confirmed by tests on the taste.

According to the present invention is also identified and proposed antagonist with a variety of antagonistic properties, which can be used in foods, beverages, Lek is stannah tools and other substances for ingestion by humans or animals, containing known and unknown bitter compounds, in which the bitter taste appropriately minimized or eliminated.

The fact that the above taste buds in particular respond to bitter connection (connection), present in coffee, and in particular bitter compounds that interact with one, multiple, or unknown receptor bitter taste, was determined essentially by the system for the expression of HEK293 and calcium imaging methods described in other publications, and patent applications filed by the present assignee, for example, U.S. ser. No. 10/191058 and 09/825882, both of which are fully incorporated into this description by reference. More specifically, the present inventors have transfusional HEK293 cells specific hT2R labeled rodopoulou label 35 amino acids (SEQ ID NO:1), together with the chimeric G-protein (G16gust44), which contains the sequence of a G protein Gα16that is modified by the replacement of the carboxy-44 amino acid residues on the remains of gustducin, and recorded the reaction of these cells to specific bitter ligands using calcium imaging methods.

In particular, the inventors used the analysis on the basis of mammalian cells for monitoring the activity of the hT2R. For calcium imaging assays cell height is ivali in 48-well plates to tissue culture. After 24 hours, cells were temporarily transfusional the expression plasmid (pEAK10)containing the nucleic acid sequence hT2R, and plasmid (pEAK10)containing a chimeric G protein (G16gust44). Even after 24 hours, cells were incubated with a fluorescent dye, specific calcium (Fluo-4; Molecular Probes). Loaded cells exposed to various bitter molecules, and activation hT2R leads to activation G16gust44, which in turn leads to mobilization of calcium inside the cells. This increase in calcium concentration changes in fluorescent properties of the calcium dye inside the cells. These changes are controlled by fluorescence microscopy.

The inventors have also used an automated fluorimetric sighting system FLIPR using a slightly different Protocol. Line HEK293 cells, stably expressing G16gust44, transfusional the expression plasmid hT2R, after 24 hours, cells were loaded and analyzed on FLIPR.

After identification of the ligand to the subject specific hT2R receive line HEK293 cells, stably expressing how hT2R and G16gust44, facilitating subsequent screening assays to identify other ligands that activate specific hT2R or which modulate (inhibit or enhance) the activation hT2R with another bitter ligand, Taco is about as bitter compound, contained in coffee. This allows you to avoid transient transfection.

As shown in the figures, as a result of these experiments it was revealed that hT2R8 and hT2R14 react to present in coffee, bitter compounds, and identified compounds that inhibit or block the bitter taste of coffee. In experiments 5 and in Example 3 below identifies the main antagonistic Connection properties, in particular.

These results show that cells that have identified taste receptors hT2R, can be used in assays to identify ligands for modulation of the bitter taste associated with at least one of these specific hT2R, as well as in assays to detect compounds that are responsible for the bitter taste.

Preferably, these analyses will be used to test cell which expresses DNA encoding hT2R containing one of the amino acid sequence identified below. However, it is expected that the fragments orthologues, variants or chimeras polypeptides of these receptors, which retain the functional properties of these receptors bitter taste, i.e. react to some bitter compounds will also be useful in these assays. Examples of such variants include splice variants, single nucleotide polymorphisms, allelic variations who you and mutation, the obtained recombinant or chemical methods or found in nature. Methods of isolation and expression of receptors T2R, which are used in the analyses according to the present invention and analyses, which are provided for use in the present invention to identify compounds that inhibit the activation of these receptors, below.

Isolation and expression of T2R

Isolation and expression of T2R or their fragments or variants according to the present invention can be implemented using conventional cloning methods using samples or primers designed based on the sequences of nucleic acids T2R presented in this application. Sequences related T2R, you can also identify the databases of the human genome and other species using the sequences disclosed in the present description and known computer search technologies, for example, the search program sequences BLAST. In one embodiment, pseudogenes described in this application can be used for identification of functional alleles or related genes.

Vectors expresii then you can apply for infection or transliterowany in cell host for functional expression of the target sequences. These genes and in story you can create and Express in vitro or in vivo. The person skilled in the art will understand that the phenotypes suitable for changing and controlling the expression of nucleic acids can be obtained by modulating the expression or activity of genes and nucleic acids (for example, promoters, enhancers, etc) in the composition of the vectors according to the present invention. You can apply any of the known methods described decrease or increase the expression or activity. The present invention can be used in combination with any of the methods or protocols known in the field, and is described in detail in the scientific and patent literature.

Also nucleic acids can be synthesized in vitro using well-known methods of chemical synthesis, as described in, for example, Carruthers, Cold Spring Harbor Symp. Quant. Biol. 47:411-18 (1982); Adams, Am. Chem. Soc., 105:661 (1983); Belousov, Nucleic Acids Res. 25:3440-3444 (1997); Frenkel, Free It. Biol. Med. 19:373-380 (1995); Blommers, Biochemistry 33:7886-7896 (1994); Narang, Meth. Enzymol. 68:90 (1979); Brown, Meth. Enzymol. 68:109 (1979); Beaucage, Tetra. Lett. 22:1859 (1981); U.S. patent No. 4458066. Double-stranded DNA fragments can be obtained by synthesizing a complementary chain and annealing of chains under appropriate conditions or by adding the complementary chain using DNA polymerase with an appropriate sequence of primer.

Methods of manipulation of nucleic acids, for example, to obtain mutations in the sequence is uncloneable, making labels in the sample, sequencing, hybridization, and the like, are described in detail in the scientific and patent literature. See, for example, Sambrook, ed., Molecular Cloning: A Laboratory Manual (2nd ed.), Vols. 1-3, Cold Spring Harbor Laboratory (1989); Ausubel, ed., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., New York (1997); Tijssen, ed., Laboratory Techniques in Biochemistry and Molecular Biology: Hybridization With Nucleic Acid Probes, Part I : Theory and Nucleic Acid Preparation, Elsevier, N.Y. (1993).

Nucleic acids, vectors, capsid, polypeptides, similar can be analyzed and quantified by any of the many conventional methods, well known to specialists in this field. Such methods include, for example, methods of analytical biochemistry, such as NMR, spectrophotometry, radiography, electrophoresis, capillary electrophoresis, high performance liquid chromatography (HPLC), thin layer chromatography (TLC), and hyperdiffusion chromatography, various immunological methods, for example, the reaction of precipitation in gel or liquid, immunodiffusion, immunoelectrophoresis, radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, southern blot, Northern blot, dot blot, gel electrophoresis (e.g., polyacrylamide gel electrophoresis with sodium dodecyl sulfate), PCR real-time quantitative PCR, other methods of nucleic acid amplification, target or signal amplification, in the Eseniya a radioactive label, pulse counting, and affinity chromatography.

For amplification of the nucleic acid encoding the binding site of the ligand T2R, you can apply oligonucleotide primers. Nucleic acids described in the present application, it is also possible to clone and to determine their number using amplification techniques. Methods of amplification are also well known in the field and include, for example, polymerase chain reaction (PCR) (Innis ed., PCR Protocols, a Guide to Methods and Applications, Academic Press, N.Y. (1990); Innis, ed., PCR Strategies, Academic Press, Inc., N.Y. (1995)); ligase chain reaction (LCR) (Wu, Genomics, 4:560 (1989); Landegren, Science, 241:1077 (1988); Barringer, Gene, 89:117 (1990)); transcription amplification (Kwoh, PNAS, 86:1173 (1989)); self-sustained replication sequences (Guatelli, PNAS, 87:1874 (1990)); Qβ replicato amplification (Smith, J. Clin. Environ., 35:1477-91 (1997)); automated amplication analysis replicate Qβ (Burg, Mol. Cell. Probes, 10:257-71 (1996)); and other methods using RNA polymerase (for example, first NASBA, Cangene, Mississauga, Ontario). Cm. also, Berger, Methods Enzymol., 152:307-16 (1987); Sambrook; Ausubel; U.S. Pat. Nos. 4,683,195 and 4,683,202; Sooknanan, Biotechnology, 13:563-64 (1995).

Amplificatoare nucleic acid, either separately or in the form of libraries, if needed, can be cloned according to methods known in this field, in any of a number of vectors using conventional molecular biology techniques; methods Cloner is of amplified nucleic acids in vitro described, for example, in U.S. patent No. 5426039. To facilitate cloning of the amplified sequences, a pair of primers for PCR can be embedded sites enzymatic restriction. For example, a pair of primers according to the present invention was built sites Pst I and Bsp E1. These special restriction sites contain the sequence, so if they were put to legirovanie, they will be inside reading frames with the "donor" of the coding sequence of the receptor, which consists of 7 transmembrane domains, into which their splitinput (the coding sequence of the binding site of the ligand is located inside relative to the polypeptide consisting of 7 transmembrane polypeptides, thus, if you want the design was broadcast in the forward direction from splanirovano restriction site enzyme products outside reading frames are undesirable; this may be required if the included area ligand mainly represents the transmembrane domain VII). To preserve the original sequence of the donor receptor, consisting of 7 transmembrane domains, you can create primers. Alternatively, the primers can encode amino acid residues that represent conservative substitutions (e.g., hydrophobic to hydrophobic residue, see what's above data), or functionally useful sequence (for example, non-limiting Assembly of the plasma membrane, causing cleavage by peptidases, causing incorrect fold of the receptor, and the like).

You can create a pair of primers for selective amplification of binding sites of the ligand protein family T2R. These plots of binding ligand may vary their ligands; therefore, the area that can serve as a minimal binding site for a single ligand, may be too limited for the second potential ligand. Consequently, it is possible to amplify parts of the binding T2R (with 7 transmembrane domains) of different sizes, consisting of different domains; for example, the transmembrane (TM) domains II through VII, III, VII, III, VI or II-VI or their variants (e.g., only a subsequence of a particular domain, a sequence with a changed order of the domains, and the like) or T2R, including the 7 transmembrane domains.

Since the domain structure and sequence of many proteins of the T2R family, composed of 7 transmembrane domains known specialist in this field can choose flanking and internal domain sequence as model sequences to create a degenerate primer pairs for amplificati is. For example, the sequence of the nucleic acid encoding the second and VII domains, you can create a method of amplification using PCR reaction using a pair of primers. For amplification of nucleic acid that carries the sequence I transmembrane domain (TM I), you can create a degenerate primer of the nucleic acid that encodes a 1 consensus sequence of the T2R family, described above. Such a degenerate primer can be used to create a binding site, including TM I-III, TM I to IV, TM I-V, TM I to VI or TM I-VII). Based on the consensus sequences of proteins of T2R family presented in this application, you can create other degenerate primers. Such a degenerate primer can be used to create a binding site, including TM from III to IV, with TM III through V, with TM III through VI or TM from III to VII.

Schema create pairs of degenerate primers is well known in this field. For example, the computer program COnsensus-DEgenerate Hybrid Oligonucleotide Primer (CODEHOP) is available at http://blocks.fhcrc.org/codehop.html and directly related to the website BlockMaker, performing alignment of multiple sequences to determine the hybrid primers, starting with a series of related protein sequences, such as the well known sites of binding of the ligand gustatory receptor (see, for example, Rose, Nucleic Acids es., 26:1628-35 (1998); Singh, Biotechniques, 24:318-19 (1998)).

Tools of the synthesis of pairs of oligonucleotide primers is well known in this field. You can use the "natural" base pairs or synthetic base pairs. For example, the use of artificial nucleotide bases provides optionality versatile approach to the manipulation of the sequence of the primer and the creation of a more complex mixture of amplification products. Different collections of artificial nucleotide bases capable of acquiring multiple directions of the hydrogen bonds by shifts of the internal connections for receiving neobyazatelnostyu degenerate molecular recognition. The introduction of such analogues in one position of the primers for PCR allows you to create complex library of amplification products. See, for example, Hoops, Nucleic Acids Res., 25:4866-71 (1997). To obtain structural analogues of natural reason in the structure of DNA can also be used and nonpolar molecules. Structural analogue without hydrogen bonding of adenine can successfully and selectively replicated against nonpolar structural analogue of thymine (see, for example, Morales, Nat. Struct. Biol., 5:950-54 (1998)). For example, two degenerate bases can be pyrimidine base 6H, 8H-3,4-dihydropyrimido[4,5-c][1,2]oxazin-7-one or purine base N6-methoxy-2,6-diaminopurine (see, for example the EP, Hill, PNAS, 95:4258-63 (1998)). Examples of degenerate primers according to the present invention include the analogue of a nucleotide base 5'-dimethoxytrityl-N-benzoyl-2'-deoxycytidine, 3'-[(2-cyanoethyl)-(N,N-aminobutiramida)]phosphoramidite (the term "P" in the sequences, see above). This pyrimidine analog form hydrogen bonds with the purine, including the remains of A and G.

Polymorphic variants, alleles, and interspecies homologues, is essentially identical to the receptor described in the present application, can be distinguished using a sample of nucleic acids described above. In another embodiment, cloning and polypeptides, polymorphic variants, alleles and interspecies homologs T2R you can use libraries of expressed sequences, defining expressed homologues immunologically method using antisera or purified antibodies to the T2R polypeptide, which also recognize and selectively bind homolog T2R.

Nucleic acid encoding the binding sites of ligands taste receptors can be obtained by amplification (e.g., using PCR) corresponding sequences of nucleic acids, using the appropriate (ideal or degenerate) pairs of primers. Nucleic acid, which will amplify, can be a DNA of any cell or tissue, or mRNA or cDNA obtained the th cell, expressing taste receptor.

In one embodiment, the design of hybrid sequences encoding proteins comprising a nucleic acid encoding T2R, merged with the sequences responsible for the translocation. Also proposed hybrid T2R, including the translocation sites and the sites of binding of the compounds that cause the sensation of taste, other families dynamics of receptors, in particular palate. These nucleic acid sequences can functionally associate with the control elements of transcription or translation, for example, with sequences to initiate transcription and translation, promoters and enhancers, terminators of transcription and translation, polyadenylation sequences, and other sequences involved in the process of transcription of DNA into RNA. In the construction of polygenic expressed clusters, vectors and transgenes, in order to achieve the accurate expression of a target nucleic acid in all target cells or tissues, you can use a fragment of the promoter.

In another embodiment, fusion proteins can enable C-terminal or N-terminal sequence of the translocation. Additionally, fusion proteins may contain additional elements, for example, for detection of proteins, their eyes the weave or for other purposes. Detection and purification of such subsidiary domains include, for example, the use of peptides that form complexes with metals such as polyhistidine channels, histidine-tryptophan elements, or other domains that allow purification on immobilized metals; maltose-binding protein; protein a domains that allow purification on immobilized immunoglobulin; or the domain used in the purification system, based on the use of the label FLAG and affinity (Immunex Corp, Seattle Wash.).

The inclusion of degradable linker sequences such as Factor Xa (see, for example, Ottavi, Biochimie, 80:289-93 (1998)), the motif of recognition subtilisin protease (see, e.g., Polyak, Protein Eng., 10:615-19 (1997)); enterokinase (Invitrogen, San Diego, Calif.), and the like, between the translocation domain (for sufficient expression of the plasma membrane) and the rest of the translated peptide, it may be useful to facilitate purification. For example, one structure may include a polypeptide encoding nucleic acid sequence linked to six histidine residues followed by thioredoxin, the site of cleavage (restriction) for enterokinase (see, for example, Williams, Biochemistry, 34:1787-97 (1995)), and C-terminal translocation domain. Residues of histidine facilitate the detection and purification, while the AIT splitting (cm) for enterokinase provides optionality clean target(s) protein(s) from the remaining protein fusion. Technology related vectors encoding fusion proteins, and methods of using the proteins of the merger, described in detail in the scientific and patent literature (see, for example, Kroll, DNA Cell. Biol,. 12:441-53 (1993)).

The expression vectors, as a separate expression vectors, and their libraries, comprising sequences encoding the binding site of the ligand can be incorporated into the genome, or in the cytoplasm or in the nucleus of the cell, and to Express by using many conventional techniques are described in detail in the scientific and patent literature. See, for example, Roberts, Nature, 328:731 (1987); Berger, supra; Schneider, Protein.. Purif., 6435:10 (1995); Sambrook; Tijssen; Ausubel. Technical descriptions provided by the manufacturers of biological reagents also contain information regarding known biological methods. Vectors can be isolated from natural sources, obtained from such sources as ATCC or GenBank library, or obtain by using synthetic or recombinant methods.

Nucleic acids can be Express in polygenic clusters, expression vectors or viruses, permanently or temporarily expressed in cells (e.g., epilimnia expression systems). To make the cells and the sequences subjected to transformation phenotype, providing optionality selection, polygenic clusters and expression vectors can include Mar the career selection. For example, the marker selection can encode information about maintaining episomal replication, eliminating the need to integrate into the genome of the host cell. For example, the marker may encode resistance to antibiotics (such as chloramphenicol, kanamycin, G418, bleomycin, hygromycin) or resistance to herbicides (for example, chlorsulfuron or Basta) for selection of cells that have been subjected to transformation target DNA sequences (see, for example, Blondelet-Rouault, Gene, 190:315-17 (1997); Aubrecht, J. Pharmacol. Exp. Ther., 281:992-97 (1997)). Since genes markers selection, giving resistance to such substrates as neomycin or hygromycin, can only be used in tissue culture, as markers for selection in vitro and in vivo also use genes chemoresistance.

The chimeric nucleic acid sequence can encode the binding site of the ligand T2R within any of the polypeptide consisting of 7 transmembrane domains. Due to the fact that the polypeptide receptors, consisting of 7 transmembrane domains have the same primary sequence and the secondary and tertiary structure, structural domains (e.g., the extracellular domain, transmembrane the domain, cytoplasmic domain, and so on) can be identified by sequence analysis. For example, using homological modeling, the Fourier analysis and determining the periodicity of the helix can identify and describe the seven domains with sequence the corresponding receptors, consisting of 7 transmembrane domains. For the assessment of prevailing periods characterizing the hydrophobicity profiles and variability of the analyzed sequences, we can apply the algorithm of fast Fourier transform (FFT). Improved methods of determining the frequency and coefficient frequency of alpha-helix can provide, for example, methods according to Donnelly, Protein Sci., 2:55-70 (1993). Other alignment algorithms and modeling are well known in the art (see, for example, Peitsch, Receptors Channels, 4:161-64 (1996); Kyte &Doolittle, J. Md. Biol., 157:105-32 (1982); and Cronet, Protein Eng., 6:59-64 (1993).

The present invention includes not only the molecules of nucleic acids and polypeptides having specific sequences of nucleic acids and amino acids, but also fragments thereof, particularly fragments consisting of, for example,, 40, 60, 80, 100, 150, 200 or 250 nucleotides, or more, as well as fragments of polypeptides consisting of, for example,, 10, 20, 30, 50, 70, 100 or 150 amino acids, or more. The nucleic acid fragments do not necessarily encode an antigenic polypeptide that is capable of contacting the antibody to the receptor of the T2R family. Next, a fragment of the protein according to the present invention can be antigenic fragment able to bind with the antibody to the receptor of the T2R family.

Also proposed chimeric proteins consisting of at m the d 10, 20, 30, 50, 70, 100, or 150 amino acids, or more, at least one of the T2R polypeptides described in the present application, in pairs associated with additional amino acids, representing another GPCR or part thereof, preferably a member of a superfamily of proteins that contain 7 transmembrane domains. Described chimeras can be obtained from the receptors according to the present invention and another GPCR, or they can be created by combining two or more of the presented receptors. In one embodiment, one portion of the chimeric protein match, or it is derived from the transmembrane domain of the T2R polypeptide according to the present invention. In another embodiment, one portion of the chimeric protein corresponds to, or receive from one or more transmembrane fragments of a T2R polypeptide, described in this application, and the remaining part or parts can be obtained from another GPCR. Chimeric receptors are well known in this field and are also well-known techniques for their creation and selection, as well as boundaries of domains or fragments of receptors associated with G-protein (GPCR). Thus, the person skilled in the art can apply this knowledge to create such chimeric receptors. The use of such chimeric receptors can provide a requirement, for example, to describe the taste specification is of one of the receptors, described in this application, as well as to characterize the transmission of the signal through another receptor, such as is well known, the receptor used in the methods of analysis according to the prior art.

For example, such a plot as the binding site binding ligand, extracellular domain, transmembrane domain, cytoplasmic domain, N-terminal domain, C-terminal domain, or any combination of them, can be covalently linked to a heterologous protein. For example, the transmembrane region T2R may be linked to the transmembrane region of the heterologous GPCR, or the extracellular domain of a heterologous GPCR may be associated with the transmembrane region T2R. Other heterologous proteins, among which you can choose include, for example, green fluorescent protein, polypeptides, beta-galactosidase, glutamate receptor, and polypeptides of rhodopsin, for example, N-terminal fragments of rhodopsin, for example, rhodopsin bull.

The present invention also encompasses the use of different host cells for the expression of T2R, their fragments or variants according to the present invention. To achieve high level expression of a cloned gene or nucleic acid, such as a cDNA molecule encoding T2R, fragments or variants according to the present invention, a specialist in this area usually the om case will subclavian target nucleic acid sequences in the expression vector, contains a strong promoter to direct the transcription terminator of transcription/translation, and, to a nucleic acid that encodes a protein, the binding site of the ribosome to initiate translation. Suitable bacterial promoters are well known in this field and are described, for example, in Sambrook et al. Preferably, expression of the receptor hT2R use of expression systems in eukaryotic cells.

For introducing foreign nucleotide sequences into a cell host, you can apply any of the well known methods. These methods include the use of calcium phosphate transfection, polybrene, fusion of protoplasts, electroporation, liposomes, microinjection, plasma vectors, viral vectors, and other well known methods for introducing cloned genomic DNA, cDNA, synthetic DNA or other foreign genetic material into the cell host (see, for example, Sambrook et al.). The only necessary condition is that the specific process of genetic design made it possible to make at least one molecule of nucleic acid in the cell-master, able to Express T2R, its fragment or a target option.

After introduction of the expression vector into the cells, transfetsirovannyh cells are cultured under conditions favoring expression of the receptor, is ragment, or the target version, which is then separated from the culture using standard techniques. Examples of such techniques are well known in this field. See, for example, WO 00/06593 placed in the present description by reference in the corresponding opening.

Methods of analysis for the detection of compounds modulating the activity hT2R according to the present invention

Below are methods and compositions for determining whether associated specific way test the connection with the T2R polypeptide according to the present invention in vitro or in vivo. To measure the impact of ligand binding chimeric T2R relatively natural, you can control many aspects of cell physiology. These methods of analysis can be performed using intact cells expressing the T2R polypeptide, cells with permeabilities membrane, or membrane fractions obtained by standard methods.

Taste receptors bind compounds that cause taste, and initiate the conversion of chemical stimuli into electrical signals. Activated or inhibited G-protein is, therefore, to modify the properties of the enzyme targets, channels and other proteins and effectors. Some examples are the activation of cGMP phosphodiesterase by transducin in the visual system, adenilatziklaznuu G-protein, phospholipase C Gq and other G-proteins, and modulation of various channels Gi and other G-proteins. You can also assess the impact, such as the production of diacylglycerol and IP3 by phospholipase C, and, further, the mobilization of calcium IP3.

Proteins hT2R, which are the objects of study, usually chosen from the polypeptides having the sequence shown in the list of sequences in the present description before formula, or its fragments or variants, the resulting conservative modifications.

Alternatively, the proteins or polypeptides T2R for analysis can be obtained from the eukaryotic host cell, and they can contain an amino acid sequence having a certain percentage identity to the polypeptides hT2R, or conservatively modified variants. Usually, the identity of the amino acid sequence comprises at least 30%, preferably 30-40%, more preferably 50-60, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. Proteins or polypeptides T2R for analysis is not necessarily contain any stretch of the T2R polypeptide, such as an extracellular domain, transmembrane domain, cytoplasmic domain, the binding site of the ligand, etc. are not Necessarily shown in the present description as an example of a T2R polypeptide or part thereof, may be covalently linked to a heterologous protein by the research chimeric protein, used in the methods of analysis described in this application.

Modulators T2R activity can be analyzed with the use of proteins or T2R polypeptides described above, as recombinant and natural. Proteins or polypeptides T2R you can select, to Express in the cells that Express the membrane isolated from a cell, to Express in tissue culture or in an animal's body, both recombinant and natural. For example, you can use slices of tongue, cells isolated from the language, transformed cells, or membranes. Modulation can be analyzed by one way analysis of in vitro or in vivo, described in this application.

The detection of modulators

Compositions and methods for determining in vitro and in vivo, binds investigated whether the connection-specific way T2R receptor according to the present invention, is described below. To measure the binding of ligand to the T2R polypeptide according to the present invention can control many physiological parameters of cells. Describes how analysis can be performed on intact cells expressing the dynamics of the receptor cells with permeabilities membrane, or membrane fractions obtained by standard methods, or in vitro, using proteins synthesized de novo.

In vivo, taste buds contact with what disiniame, modulating the taste, and initiate the transduction of chemical stimuli into electrical signals. Activated or inhibited G-protein is, therefore, to modify the properties of the enzyme targets, channels and other effector proteins. Some examples are the activation of cGMP phosphodiesterase by transducin in the visual system, adenylate cyclase stimulatory G-protein, phospholipase C Gq and other G-proteins, and modulation of various channels Gi and other G-proteins. You can also analyze further the effects on parameters such as the production of diacylglycerol and IP3 by phospholipase C, followed by mobilization of calcium IP3.

Alternatively, the proteins or polypeptides T2R used in the analysis, can be obtained from eukaryotic host cells, and they can include an amino acid sequence with an identity of amino acids with T2R polypeptides described in this application or their fragments, or variants, obtained by conservative modifications. Usually the identity of the amino acid sequence is at least 35 to 50%, or optional, 75%, 85%, 90%, 95%, 96%, 97%, 98% or 99%. Optional, proteins or T2R polypeptides used in the methods of analysis may consist of domain T2R protein, such as an extracellular domain, a transmembrane domain, transmembrane region, cytoplasmatic the cue domain, the binding site of the ligand, and the like. Further, as described above, T2R protein or its domain can be covalently to contact heterologous protein with the formation of a chimeric protein used in the methods of analysis described in this application.

Modulators of the activity of the receptor T2R investigate, using proteins or polypeptides T2R, as described above, both recombinant and natural origin. Proteins or polypeptides T2R you can select, to Express in the cell to Express the membrane isolated from a cell, to Express in the tissue or the body of the animal, as recombinant or natural origin. For example, you can use slices of tongue, cells isolated from the language, transformed cells, or membranes. Modulation can be investigated by applying one of the described in the present application methods in vitro or in vivo.

Analyses of binding in vitro

The taste transduction can also be examined in vitro in reactions in the solid or liquid phase using T2R polypeptides according to the present invention. In one embodiment, the binding site of the ligand T2R can be used in reactions in vitro, in solid or liquid phase for analysis of ligand binding.

Optional receipt of the binding site of the ligand of the N-terminal region with additional parts of the extracellular domain, such as wncl the exact loops of the transmembrane domain.

Also carried out the analysis of binding in vitro with other GPCR, such as metabotropic glutamate receptors (see, for example, Han and Hampson, J. Biol. Chem. 274:10008-10013 (1999)). Such analyses may include the adoption of a ligand with a radioactive or fluorescent label, the measurement of changes in the characteristic fluorescence or changes in proteolytic sensitivity, etc.

The binding of ligand to the T2R polypeptide according to the present invention can be explored in solution, on a double-layer membrane, optionally immobilized on a solid phase, in a lipid monolayer, or in liposomes. Binding of the modulator can be investigated by measuring for example, changing spectroscopic (e.g., fluorescence, absorbance, refractive index), hydrodynamic (e.g., shape), chromatographic characteristics, or properties of solubility.

In a preferred embodiment of the invention, applied to the analysis of binding[35S]GTPγS. As described above, in relation to activation of the GPCR, the Gα subunit of the G-protein stimulates conversion of HDF in the GTP. The stimulation of metabolic activity of G-protein-mediated ligand can be measured using biochemical methods of analysis, by assessing binding[35S]GTPγS with a radioactive label and G-protein in the presence of presumptive ligand. Usually membrane, stereoselective dynamics of the receptor, mixed with G-protein. Potential inhibitors and/or activators and[35S]GTPγS added to the reaction mixture, and evaluate binding[35S]GTPγS G-protein. Binding can be assessed by scintillation counting fluid, or by any other method known in this field, including scintillation analysis of approximation (SPA). In other methods of analysis can be used GTPγS labeled.

Fluorescence polarization analysis

In another embodiment, for detection and control of ligand binding is possible to apply methods of analysis based on fluorescence polarization ("OP"). Fluorescence polarization is a universal laboratory technique for measurement of equilibrium binding, hybridization of nucleic acids, and enzymatic activity. Fluorescence polarization assay is a homogeneous, as it does not require phase separation, such as centrifugation, filtration, chromatography, precipitation or electrophoresis. This analysis is performed in real time, directly in solution without phase immobilization. The values of polarization can be evaluated again after adding reagents, as the measurement of polarization occurs quickly and does not destroy the sample. In General, these techniques can be used to measure values of polarization of the fluorophores in the range is from low picomolar to micromolar levels. In this section, the optional use of fluorescence polarization in a simple and quantitative analysis of the sample to assess the binding of ligands with T2R polypeptides according to the present invention.

Upon excitation of the molecule with a fluorescent label using linear plane-polarized light, it emits light, the degree of polarization of which is inversely proportional to its molecular rotation. Large molecules with a fluorescent label remain relatively stationary during the stage of initiation (in the case of fluorescein, is 4 nanoseconds), and the polarization of the light remains relatively constant during the transition from the excitation radiation (emission). Small molecules with a fluorescent label slowly during the stage of excitement, and polarization changes significantly during the transition from excitation and emission. Thus, small molecules have low values of polarization, and large molecules have large values of polarization. For example, single-stranded oligonucleotide with the included fluorescent label has a relatively low value of the polarization, but by hybridization with a complementary chain is its polarization increases. When using the OP for the detection and control of the binding compounds that cause taste sensations that can activate or ing biavati dynamics of the receptors according to the present invention, it is possible to use compounds that cause taste, with a fluorescent label, or autofluorescent compounds that cause taste.

Fluorescence polarization (P) is determined as follows:

,

where Intparrepresents the intensity of light emitted parallel to the plane of the exciting light, and Intperpis the intensity of the light emitted perpendicular to the plane of the exciting light. P, which is the ratio of light intensities, is a dimensionless quantity. In combination with this analysis can be applied, for example, Beacon™ Beacon 2000™. Such systems usually expressed polarization in units of millionaireasia (1 unit polarization=1000 mP units).

The relationship between molecular rotation and size is described by the Perrin equation, see Jolley, M. E. (1991) in the Journal of Analytical Toxicology, pp. 236-240 included in the present description by reference, which describes the given equation. Briefly, according to the Perrin equation, the degree of polarization is directly proportional to the magnitude of the relaxation period, the time needed for a molecule to rotate at an angle of about 68,5°. The relaxation period of rotation is associated with the viscosity (η), absolute temperature (T), molecular volume (V) and the gas constant (R) as ur is the ranking: 2 (relaxation period of rotation)=3 V RT.

The relaxation period of rotation has small values (~ nanosecond) for small molecules (e.g. fluorescein) and large (~100 NS) for large molecules (e.g., immunoglobulins). If the viscosity and temperature constant support, the relaxation period of rotation and, accordingly, the polarization is directly proportional to the size of the molecule. Changes in molecular volume can occur due to interactions with other molecules, dissociation, polymerization, decomposition, hybridization or conformational changes of the molecule with a fluorescent label. For example, fluorescence polarization was used to assess enzymatic degradation by proteases, Dnisone and RNase large polymers with fluorescent label. Also it was used to estimate the equilibrium binding for protein-protein interactions, interactions of antigen-antibody, and the interactions protein-DNA.

High-performance methods of analysis in the liquid and in the solid phase

In yet another embodiment, the proposed analysis in the liquid phase with the use of a T2R polypeptide, or cell or tissue expressing the T2R polypeptide. In another embodiment according to the present invention is proposed performance analysis in vitro, in solid phase, in which the T2R polypeptide, or a cell or tissue expressing polypep the d T2R, fixed on the solid phase substrate or a compound that stimulates the taste, and are put in contact with the T2R receptor, binding determine using the appropriate labels or antibodies to receptor T2R.

High-performance methods of analysis according to the present invention provide a requirement to carry out screening to several thousand different modulators or ligands in a single day. In particular, each well of the microtiter tablet can be used to conduct separate analyses with selected potential modulator, or, if you examine the influence of the concentration or the incubation time, you can explore one modulator in every 5-10 wells. Thus, a single standard microtiter tablet, you can analyze about 100 (e.g., 96) modulators. If using tablets with 1536 wells, in one tablet, you can easily analyze from about 1000 to about 1500 different compounds. Also optional to successfully carry out the analysis of several compounds in each well of the plate. Not necessary to analyze multiple tablets per day; the application of integrated systems according to the present invention provides for the optional implementation of screening to approximately 6000-20000 different compounds. Were later developed the Ana microfluidic approaches for manipulation of the reagents.

The target molecule can be attached to solid state component, directly or indirectly, via covalent or non-covalent connection, for example, by using labels. The label may be any of a plurality of components. In General, a molecule that binds the label, fixed on a solid basis, and the target molecule (for example, the target molecule transduction of taste) entered into the label associated with the solid substrate by interaction of the tag and molecules that bind tag.

You can use a variety of labels and molecules that bind a label based on molecular interactions are well described in the literature. For example, if the label exists linking molecule of natural origin, for example, Biotin, protein a, or G-protein, it can be used in conjunction with the relevant molecules, binding tag (avidin, streptavidin, neutravidin, Fc fragment of immunoglobulin, etc). Antibodies to these molecules with natural binding molecule, such as Biotin, are also widely available, as appropriate molecule that binds the tag (see, SIGMA Immunochemicals 1998 catalogue SIGMA, St. Louis Mo.).

Also, to obtain a pair tag/binding label compound can be any compound heptanol or antigenic nature in combination with a corresponding antibody. Thousands of specific and which antibodies are commercially available, many other antibodies described in the literature. For example, in one conventional configuration, the label is the first antibody, and binding the label compound is a second antibody that recognizes the first antibody. In addition to the interactions of antigen-antibody interaction, receptor-ligand can also be used in pairs tag/binding tag connection. Examples are agonists and antagonists of receptors of the cell membrane (for example, such interaction cell receptor-ligand as transferrin, c-kit, viral ligands of receptors, cytokine receptors, receptors chemokines, receptors, interleukin receptors, immunoglobulin and antibodies, family catherinew, the integrin family, the family of selectins, and the like; see, e.g., Pigott &Power, The Adhesion Molecule Facts Book I (1993)). Similarly, with different receptors of the cells can interact toxins and venoms, viral epitopes, hormones (e.g., opiates, steroids, etc), intracellular receptors (e.g., which are the mediators of the effects of various small ligands, including steroids, thyroid hormone, retinoids and vitamin D; peptides), drugs, lectins, sugars, nucleic acids (both linear and cyclic polymer configurations), oligosaccharides, proteins, phospholipids and antibodies.

The appropriate label or a binding label is connected to the I can also be obtained from synthetic polymers, such as polyurethanes, polyesters, polycarbonates, polyureas, polyamides, polyethylenimine, Polyarylamide, polysiloxanes, polyimides and polyacetate. Many other couples tag/connecting the label compound is also applicable in systems analysis, described in the present application, as will be clear to the person skilled in the art from a review of this opening.

Also as targets, you can use basic linkers, such as peptides, polyethers, and the like, including a polypeptide sequence, such as polyglycine sequence length from about 5 to about 200 amino acids. Such flexible linkers known to specialists in this field. For example, poly(etilenglikolevye) linkers available in Shearwater Polymers, Inc. Huntsville, Ala. Such linkers may contain amide linkages, sulfhydryl communication, or heterofunctional connection.

Connecting the connection mark immobilized on solid substrates using any of the many currently available methods. Solid substrates usually get or functionalitywith through the implementation of the interaction of the substrate, in whole or in part with a chemical reagent, fixing chemical group on the surface, reacting with a portion connecting the connection mark. Examples of groups suitable for attachment to a long chain which include amine, hydroxyine, tirinya and carboxyl groups. For functionalliteracy various surfaces, such as glass surface, you can apply aminoalkylsilane and hydroxylysine. The receipt of such solid phase biopolymer structures are well described in the literature. See, for example, Merrifield, J. Am. Chem. Soc., 85:2149-2154 (1963) (describing solid phase synthesis, for example, peptides); Geysen et al., J. Immun. Meth., 102:259-274 (1987) (describing synthesis of solid phase components on pins); Frank &Doring, Tetrahedron, 44:60316040 (1988) (describing synthesis of various sequences of peptides on cellulose disks); Fodor et al., Science, 251:767-777 (1991); Sheldon et al., Clinical Chemistry, 39(4):718-719 (1993); and Kozal et al., Nature Medicine, 2(7):753759 (1996) (all materials listed above describe the synthesis of biopolymers attached to solid substrates). Non-chemical approaches to attaching the binding label of compounds to substrates include conventional methods such as heating, the formation of cross-links under the action of UV radiation, etc.

Cellular analyses

In one of the preferred embodiments T2R protein Express in eukaryotic cells as in unmodified form or in the form of a chimeric modified or truncated receptor with or preferably without heterologous sequences chaperone, promoting its maturation and targeting to the secretory pathway. That is their T2R polypeptides can be Express in any eukaryotic cell, such as cells HEK-293. In the preferred case, the cells contain a functional G protein, e.g., G.α15, or chimeric G.α16, gustducin or transducin, or chimeric G-protein, such as G16gust44 able to contact chimeric receptor intracellular signaling pathway or to a signaling protein such as phospholipase C. Activation of T2R receptors in such cells can be determined using any standard method, such as determining changes in the content of intracellular calcium using FURA-2 dependent fluorescence in the cell. The described analysis is the basis of the experimental results presented in this application.

Activated GPCR receptors are often substrates for kinases, fosfauriliruetsa C-terminal fragment of the receptor (and optionally, also other sites). Thus, activators help to transfer with 32P labeled ATP receptor, which can be measured using a scintillation counter. Phosphorylation of the C-terminal end will contribute to the binding of arrestin-like proteins, and will prevent the binding of G-proteins. A General overview of GPCR signal transduction and analysis of signal transduction is shown, for example, Methods in Enzymology, vols. 237 and 238 (1994) and volume 96 (1983); Bourne et al., Nature, 10:349:117-27 (1991); Bourne et al., Nature, 348:125-32 (1990); Pitcher et al., Annu. Rev. Biochem., 67:653-92 (1998).

Modulation T2 can be investigated by comparing the response of a T2R polypeptide, processed putative modulator T2R, answer untreated control sample or a sample containing a known positive control. Such estimated T2R modulators may include molecules that either inhibit or activate the activity of a T2R polypeptide. In one embodiment, the control samples treated with compound that activates T2R, assign relative T2R activity value of 100. Inhibition of the T2R polypeptide reach when T2R activity value relative to the control sample is 90%, optionally 50%, optionally 25-0%. Activation of T2R polypeptide reach when T2R activity value relative to the control is 110%, optionally 150%, 200-500%, or 1000-2000%.

Changes in ion flux may be measured by determining changes in polarization of the ions (i.e. electric potential) of the cell or membrane expressing the T2R polypeptide. One way to determine changes in cellular polarization is by measuring changes in current (measuring changes in polarization) with the help of fixation potential methods of registration capacity (volt-clamp) and patch-clamp (see, for example, the method "attached cells", the method "inside out", and the "whole cell", for example, Ackerman et al., New Engl. J Med., 336:1575-1595 (1997)). Currents of whole cells successfully measured using the standard and requirements. Other known methods of analysis include studies of radiolabelled ion currents using dyes that are sensitive to electric voltage (see, for example, Vestergarrd-Bogind et al., J. Membrane Biol., 88:67-75 (1988); Gonzales & Tsien, Chem. Biol., 4:269-277 (1997); Daniel et al., J. Pharmacol. Meth., 25:185-193 (1991); Holevinsky et al., J. Membrane Biology, 137:59-70 (1994)).

The influence of the studied compounds on the function of polypeptides can be determined by evaluation of any of the parameters described above. To measure the influence of the tested compound on the polypeptides according to the present invention can be measured by any suitable physiological change in the activity of GPCR. When determining the physiological effects on intact cells or animals, one can also measure effects such as the secretion of transmitter, the secretion of hormones, changes in transcription as well known and not described genetic markers (e.g., assays, Northern blot), measurement in cell metabolism such as cell growth and pH changes, and changes in the content of intracellular second messengers, such as Ca2+, IP3, cGMP, or camp.

Preferred methods for analysis of GPCR include cells loaded with dyes that are sensitive to ions or electric voltage, to describe the activity of the receptors. In the methods of analysis for determining the activity of such receptors also is possible to apply known agonists and antagonists for other pair associated with G-protein receptors as a control for measuring the activity of the investigated compounds. In the methods of analysis to identify modulating compounds (e.g., agonists, antagonists), monitor changes in the level of ions in the cytoplasm or electrical voltage on the membrane using a fluorescent indicator that is sensitive to ions or electrical stress on the membrane, respectively. The indicators that are sensitive to ions, and the sample values of an electrical voltage that can be used are described in Molecular Probes 1997 Catalog. For receptor pair associated with G-protein, the chosen method of analysis it is possible to use a random G-proteins, such as Gα15 and Gα16 (Wilkie et al., Proc. NAT'l Acad. Sci., 88:10049-10053 (1991)). Alternatively, you can use other G-proteins, such as gustducin, transducin, and chimeric G-proteins, such as Gα16gust44 or Galpha16t25.

Activation of the receptor initiates subsequent intracellular events, such as the increase in the content of secondary messengers. Activation of some receptors associated with G-proteins, stimulates the formation of Inositol triphosphate (IP3) through the hydrolysis of phosphatidylinositol, mediated by phospholipase C (Berridge &Irvine, Nature, 312:315-21 (1984)). IP3 in turn, stimulates the release of calcium from intracellular stores. Thus, changes in the level of calcium ions in the cytoplasm or changes in the levels of second messengers such as IP3, can be used on lesofat to assess the functions of receptors, associated with G-proteins. Cells expressing such receptors, G-proteins coupled, can detect elevated levels of cytoplasmic calcium, which are the result of the exit of calcium from endocellular depots and entry of extracellular calcium through ion channels in the plasma membrane.

In a preferred embodiment, the activity of the T2R polypeptide is measured by the T2R gene expression in heterologous cell with any G-protein that links the receptor transmitted signal involving phospholipase C (see Offermanns &Simon, J. Biol. Chem., 270:15175-15180 (1995)). Preferably, the cell line is HEK-293 (OK not expressing genes T2R), and G-protein is Gα15 (Offermanns &Simon, supra), or chimeric G-protein, such as Gα16gust44. Modulation of the transmission of taste signals are examined by measuring changes in levels of intracellular Ca2+that occur in response to the modulated transmission signal T2R with the introduction of molecules that communicates with the T2R polypeptide. Changes in the levels of Ca2+ can be measured using fluorescent dyes as indicators of Ca2+ and methods for fluorescence images.

In another embodiment, analyze hydrolysis phosphatidylinositol (PI) in accordance with U.S. patent No. 5436128 included in the present description by reference. Briefly, the analysis includes the introduction is tion in cell labels 3H-myoinositol 48 hours or more. Cells with introduced them in the label of the process of the studied compound in an hour. The treated cells are lysed and release in an aqueous solution of chloroform-methanol, after which inositolfosfatov separated by the method of ion exchange chromatography, and register the scintillation. The frequency of stimulation is determined by calculating the relationship cpm (counts per minute) in the presence of agonist, to the cpm in the presence of buffer control. Similarly, the rate of inhibition is determined by calculating the relationship cpm in the presence of agonist, to the cpm in the presence of buffer control (which may contain or not contain the agonist).

Other methods of analysis of receptors may involve determining the level of intracellular cyclic nucleotides, e.g., camp or cGMP. In cases where activation of the receptor leads to reduced levels of cyclic nucleotides, it may be preferable to subject the cells to exposure to agents that increase intracellular levels of cyclic nucleotides, for example, Forskolin, before adding a compound that activates the receptor to the investigated cells. In one embodiment, changes in the intracellular content of camp or cGMP can be measured using immunological assays. To determine the level of cGMP is possible to apply the method described in Offermanns &Simon, J. Bio. Chem., 270:15175-15180(1995). Also, to determine the level of cGMP is possible to apply the method described Felley-Bosco et al., Am. J. Resp. Cell and Mol. Biol., 11:159-164 (1994). Further, the kit for measuring levels of camp and/or cGMP is described in U.S. patent No. 4115538 included in the present description by reference.

In another embodiment, it is possible to measure the levels of transcription to assess the impact of the compounds on the signal transmission. Cell host containing the target polypeptide T2R lead in contact with the test compound for a time sufficient for it to have influenced the interaction, and then measure the level of gene expression. Sufficient time to effect any interactions, can be determined empirically, for example, by determining the temporal relationships and measuring the level of transcription, depending on time. The transcription level can be measured using any method known to specialists in this field. For example, the expression of the target protein mRNA can be determined by means of Northern blot, or can also be identified polypeptide products using immunological assays. Alternatively, you can apply the analysis of transcription using a reporter gene, as described in U.S. patent No. 5436128 included in the present description by reference. Reporter genes may represent,for example, genes chloramphenicol, acetyltransferase, luciferase, beta-galactosidase, beta-lactamase and alkalinity. Next, the target protein can be used as an indirect reporter via attachment to the secondary mount, such as green fluorescent protein (see, for example, Mistili & Spector, Nature Biotechnology, 15:961-964 (1997)).

Then the transcription level compared with the level of transcription in the same cell in the absence of the compounds, or it can be compared with the level of transcription in approximately identical cell that lacks(s) target(s) polypeptide(s). Essentially identical to the cell can be obtained from the same cells, which are recombinant cell, but not modified by the incorporation of heterologous DNA. Any difference in the level of transcription indicates that the test compound to some extent modifies the activity of the target polypeptide T2R.

Transgenic animals other than man, the body dynamics expressed receptors

Animals (not related to the mind of man), in the body which are expressed one or more sequences of receptors according to the present invention can also be used for receptor analysis. This expression can be used to determine whether associated investigational compound specific the way with a transmembrane complex gustatory receptor mammals in vivo, by bringing into contact of the body of the animal (non-human), stable or temporarily transfitsirovannykh nucleic acids encoding the dynamics of the receptor or binding site of the ligand of these receptors with a test compound, and determining, if an animal reacts to a monitored connection-specific binding polypeptide receptor complex.

Animals, transfetsirovannyh or infected vectors according to the present invention is particularly useful for methods of analysis for the identification and study of gustatory stimuli which are able to bind with specific receptor sites. Such animals infected with the vector expressing the sequence of taste receptors may be useful in screening in vivo gustatory stimuli and their effects, for example, at the cellular physiology (e.g., gustatory neurons in the Central nervous system or behavior.

Means for infection/expression of nucleic acids and vectors, both individually and as part of the libraries are well known in this field. Multiple parameters of individual cells, organs or whole organisms of the animal can be measured by various means. Sequence T2R according to the present invention can, for example, to Express in taste tissues of the animal, through the m introduction using the infectious agent, for example, adenoviral expression vector.

Genes endogenous taste receptor can retain the functional activity and the activity of wild-type (natural). In other cases, when it is desirable that all the functional activity of the receptor was reported he introduced exogenous hybrid receptor, it is preferable to use a line with a knockout. How to create transgenic animals (not related to the mind of man), in particular transgenic mice, as well as the selection and creation of recombinant constructions to obtain transformed cells, are well known in this field.

The construction of "knockout" cells and animal based on the assumption that the level of expression of a particular gene in a cell of a mammal can reduce or completely eliminate the expression by introducing into the genome of a new DNA sequence that serves to interrupt part of the DNA sequence that you want to suppress. Insert gene trap can also be used for the destruction of the gene of the host, and to receive the "knock-out" transgenic animals can be used stem cells from mouse embryos (ES) (see, for example, Holzschu, Transgenic Res 6:97-106 (1997)). Insert aksogan usually carried out using homologous recombination between complementary nucleotide sequences. Exogenous for sledovatelnot represents the portion of the target genes modification, such as an exon, intron, or sequences that regulate transcription, or any sequence of the genome that may affect the level of expression of the target genes, or a combination thereof. Targeting genes by homologous recombination in multipotent stem cells of the embryo allows you to modify the target sequence of the genome with high precision. For creating, screening, breeding "knock-out" animals can be applied to any technique, see, for example, Bijvoet, Hum. Mol. Genet. 7:53-62 (1998); Moreadith, J. Mol. Med. 75:208-216 (1997); Tojo, Cytotechnology 19:161 to 165 (1995); Mudgett, Methods Mol. Biol. 48:167-184 (1995); Longo, Transgenic Res. 6:321-328 (1997); U.S. patent№ 5616491; 5464764; 5631153; 5487992; 5627059; 5272071; WO 91/09955; WO 93/09222; WO 96/29411; WO 95/31560; WO 91/12650.

Nucleic acids according to the present invention can also be used as reagents for producing "knock-out" of human cells and their proginov. In this way the nucleic acid according to the present invention can be used as reagents to obtain the "products activate" (knock-in) mouse. Orthologues T2R in the mouse genome can be replaced by T2R human or rat. In this way we obtain a mouse expressing T2R human or rat. Then this mouse can be used to analyze functions T2R humans and rats, and identificirovat ligands for such T2R.

Modulators

Compounds tested as m is of deleterow receptors of the T2R family, can be any small chemical compound, or biological entity, such as a protein, sugar, nucleic acid or lipid. Alternatively, modulators can be genetically modified variants of the T2R family member. Typically, test compounds are small chemical molecules or peptides. In principle, as a potential modulator or ligand in the methods of analysis according to the present invention can be used any chemical compound, although the most frequently used compounds dissolved in aqueous or organic (especially on the basis of DMSO, DMSO) solvent. Can be designed for the analysis of extensive screening of chemical libraries by automating the analysis and providing compounds from any suitable source, for tests, usually carried out in parallel (e.g., in microtiter formats or the microtiter plates in robotic assays). The person skilled in the art it is clear that there are many suppliers of chemical compounds, including Sigma (St. Louis, Mo.), Aldrich (St. Louis, Mo.), Sigma-Aldrich (St. Louis, Mo.), Fluka Chemika-Biochemica Analytika (Buchs, Switzerland), etc.

In one embodiment, the methods of high-performance screening analysis include providing a combinatorial library chemically the substances or peptides, containing a large number of potential therapeutic compounds (potential modulator or ligand). Such "combinatorial chemical libraries, or libraries of ligands" are then subjected to screening in one or more of the methods of analysis described in this application, to identify members of the library (individual species or subclasses of chemical compounds), with the target activity. Compounds identified in this way can serve as conventional "lead compounds" or can be used as potential or actual consumption.

A combinatorial library of chemical substances is a collection of various chemical compounds obtained as by chemical synthesis, and biological synthesis, by combining a number of chemical "building blocks", such as chemical compounds. For example, a linear combinatorial chemical library such as the library of polypeptides, obtained by combining a number of chemical building blocks (amino acids) in any way possible, to obtain the compounds of a given length (i.e. the number of amino acids in the polypeptide). Through such combinatorial mixing of chemical building blocks it is possible to synthesize millions of chemical compounds.

Receiving the s and screening of combinatorial chemical libraries is widely known to experts in this field. Such combinatorial chemical libraries include, but are not limited to, peptide libraries (see, for example, U.S. patent No. 5010175, Furka, Int. J. Pept. Prot. Res., 37:487-93 (1991) and Houghton et al., Nature, 354:84-88 (1991)). To create combinatorial chemical libraries is possible to use other chemical compounds. Such chemicals include, but are not limited to, the following: peptide (for example, WO 91/19735), encoded peptides (e.g., WO 93/20242), random bialogora (for example, WO 92/00091), benzodiazepines (for example, U.S. patent No. 5288514), diversionary, such as hydantoins, benzodiazepines and dipeptides (Hobbs et al., PNAS., 90:6909-13 (1993)), vinylogous polypeptides (Hagihara et al., J. Amer. Chem. Soc., 114:6568 (1992)), nonprotein peptidomimetics with glucose frame (Hirschmann et al., J. Amer. Chem. Soc., 114:9217-18 (1992)), similar to the organic combination of small compound libraries (Chen et al., J. Amer. Chem. Soc., 116:2661 (1994)), oligocarbonate (Cho et al., Science 261:1303 (1993)), peptideprophet (Campbell et al., J. Org. Chem., 59:658 (1994)), library of nucleic acids (Ausubel, Berger, Sambrook, all, see above), the library of nucleic acids encoding peptides (U.S. patent No. 5539083), libraries of antibodies (Vaughn et al., Nature Biotechnology, 14(3):309-14 (1996) and PCT/US96/10287), library of carbohydrates (Liang et al., Science, 274:1520-22 (1996) and U.S. patent No. 5593853), libraries of small organic molecules (benzodiazepines, Baum, C&EN, Jan. 18, page 33 (1993); thiazolidinone and metatietoseminaari, Pat the t U.S. No. 5549974; pyrrolidine, U.S. patent No. 5525735 and 5519134; connections on the basis of the research, U.S. patent No. 5506337; benzodiazepines, 5288514, etc).

Equipment for creating combinatorial libraries are commercially available (see, e.g., 357 MPS, 390 MPS (Advanced Chem Tech, Louisville Ky.), Symphony (Rainin, Woburn, Mass.), 433A (Applied Biosystems, Foster City, Calif.), 9050 Plus, Millipore, Bedford, Mass.)). In addition, several commercially available combinatorial libraries (see, e.g., ComGenex, Princeton, N.J.; Tripos, Inc., St. Louis, Mo.; 3D Pharmaceuticals, Exton, Pa.; Martek Biosciences; Columbia, Md.; etc.).

According to one aspect of the present invention, T2R modulators can be used in any food product, a confectionery product, pharmaceutical composition or ingredients for directional modulation of taste of the product, composition or ingredient. For example, T2R modulators, reinforcing the sense of bitter taste, you can add to give bitter taste to the product or composition, while the T2R modulators that inhibit the sensation of bitter taste, can be added to block the bitter taste of the product or composition. Also, according to the present invention, methods of identification of bitter compounds in foods, beverages and medicines and receiving of food, beverages and medicines with improved taste, which is absent or weakened it.

The use of compounds Ident is infected according to the present invention

Compounds identified according to the present invention, can be added to foods, drinks, cosmetics or medical compositions to modulate, preferably block bitter taste caused by activation of at least one of hT2R8 and/or hT2R14 bitter compound present in coffee and coffee related products, beverages and medicines, or structurally close their connections, or other bitter compounds, for example compounds present in foods and beverages, or medicines and cosmetics, causing a feeling bitter taste.

In particular, the Connection C and its analogues, due to their properties antagonist broad-spectrum, can be used as an additive in foods, beverages, drugs, or other connection, intended for consumption by humans and animals, bitter taste which it is desirable to soften. Given the properties of this Connection, these compounds may contain bitter ligands that interact with receptors bitter taste, such as hT2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 64, 65, or 71 and/or hT2R5, 9, 13, 54, 67 and 75, or their combination, or they may contain bitter compounds, bitter taste receptors are not installed. Particularly preferred variants of PR the changes are connection, activating multiple bitter taste receptors.

In addition, the compounds according to the present invention, including Connection, can be applied in the analysis of competitive binding and functional analyses, and testing in taste samples for identification of bitter compounds, bitter taste which blocks or inhibits the Connection With.

As mentioned above, it is preferable that the ability of the compounds identified in cellular assays with T2R according to the present invention, to modulate taste, preferably block bitter taste, was confirmed through taste tests involving human or animal, preferably in a taste test involving human subjects.

Sets

T2R genes and their homologs are useful tools for identification of cells of the taste buds in forensics and paternity definition, as well as for studies of taste signaling. Reagents that are specific to members of the T2R family, subjected to specific image hybridization with nucleic acids T2R, such as samples and primers T2R, and reagents that are specific to the T2R specific way communicating with the T2R protein, for example, antibodies to T2R, used for studies of the expression of taste cells and the regulation of transmission taste (gustatory signaling).

When estva (methods) analysis of nucleic acids for the presence in the sample DNA and RNA family member T2R include numerous techniques, well-known experts in this field, such as southern analysis, Northern analysis, the method of the dot - blotting, protection from RNase, analysis S1, amplification techniques such as PCR and in situ hybridization. In the method of in situ hybridization, for example, nucleic acid, which is the target, released from its cellular environment so that it was available for hybridization within the cell, while retaining the morphology of the cells for subsequent research and analysis. The following articles contain a review of in situ hybridization: Singer et al., Biotechniques, 4:230250 (1986); Haase et al., Methods in Virology, vol. VII, 189-226 (1984); and Names et al., eds., Nucleic Acid Hybridization: A Practical Approach (1987). In addition, recombinant T2R protein can be detected using various methods of immunological analysis, described above. The test sample is typically compared with a positive control (e.g., a sample expressing recombinant protein T2R, and negative control.

According to the present invention also offered kits for screening for modulators of receptors of the T2R family. Such kits can be made from readily available materials and reagents. For example, such kits can include any of the following materials, and other materials: nucleic acids and proteins T2R, tubes for reactions, how to study the activity of T2R. Set optional is about, includes a set of functional T2R polypeptide. According to the present invention can manufacture a wide range of kits and components depending on user tasks and their specific needs.

After a General description of the invention a more complete understanding of the information provided by the following examples included in this description to illustrate, and not considered as limiting. It is obvious that described in the present application examples of embodiments of the invention it is possible to make various modifications and changes without deviating from the idea and scope of the invention.

EXAMPLES

Example 1

hT2R8 and hT2R14 activated bitter faction coffee

For screening 25 T2R person temporarily in transfected HEK cells was used partially purified fraction coffee, as described in previous patent applications. Briefly, (a more detailed description see the publication of the patent application U.S. No. 2003/0170608 included in the present description by reference), primary kidney cells (mesonephros) man, stably expressing the large T-cell antigen and protein G15 (HEK-G15) was temporarily transfusional the expression plasmid hT2R (for example, using phosphate Ca2+or systems based on lipids). Additionally, other cell line HEK-G15 was temporarily transfusional other T2R man. Then used fluorescence analysis to determine changes in concentrations of calcium in temporarily transfected cells. The interaction of test compounds (compounds) with transfitsirovannykh cells causes stimulation of the signaling cascade leading to activation of PLC and subsequent increase in the concentration of intracellular calcium, leading to the emergence of fluorescence, which was detected using a fluorescent dye that is sensitive to calcium. These changes were tracked using, for example, fluorescent microscopy and the appropriate software (such as Imaging Workstation, Axon).

Fraction coffee has a high level of fluorescence that interfere with the analysis. To overcome these obstacles have tested several blue dyes on the ability to block fluorescence fraction coffee. As shown in figure 1, the fraction coffee activated hT2R8 and hT2R14 in calcium-dependent analysis using temporarily transfected cells. In the experiment, which is illustrated by figure 1, used blue dye FD&C 1 at a concentration of 1.9 mm. Found that this fraction coffee activates several other hT2R. Using various blue dyes activated by various combinations of hT2R (table 1).However, hT2R8 and hT2R14 considered stable reacherous the mi on the fraction coffee, and activity of these two receptors depend on the number of fractions coffee (Figure 2). In the experiment, which illustrates Figure 2, used blue dye tripney blue.

Table 1
hT2R activated fraction coffee, with various blue dyes
Blue dyeIdentified receptors hT2R
activatedweakly activated
FD&C 18, 14---
Tripney1, 8, 1410, 75
Kumasi14---

When applying this method of analysis, it was found that the addition of bitter faction of coffee to cells expressing hT2R8 and hT2R14, activated by intracellular G-proteins. When applying the same method of analysis bitter faction coffee, in contrast, is not activated cells HEK-G15, which was temporarily transfusional other hT2R. This experiment confirms the conclusion that the taste buds hT2R8 and hT214 specific way respond to bitter(bitter) connection(connection), present in coffee.

Example 2

Identification of antagonists hT2R8 and hT2R14

To identify antagonists were established cell line, stably expressing hT2R8 and hT2R14, respectively, together with promiscuous chimeric protein G16g44, as described in previous patent applications. Conducted performance analysis using stable cell lines and devices FLIPR (Fluorescent Imaging Plate Reader). To activate receptors to 70-80% of their maximum relative activity used agonist hT2R8 or hT2R14. Agonist for hT2R8 was Andrographolide (200 μm); and agonist hT2R14 was Aristolochia acid (3 μm). To identify antagonists were added compounds with different chemical structure together with the agonist. Compounds that caused a statistically significant decrease in the activity of the receptor, were combined and confirmed the results with inhibition curves showing the dependence of the dose. Compound a and Compound B was identified as antagonists of hT2R8 (Figure 3). Compound C was identified as the antagonist hT2R14 (Figure 4).

Example 2a

The combination of antagonists hT2R8 and hT2R14 weaken the bitter taste of coffee

Conducted taste tests with combinations of antagonists hT2R8 and hT2R14 in the faction of coffee and two kinds of instant coffee (medium roast and roasted to medium-t is a lot of state), using the method of forced choice with 2 alternatives panel tastes involving 4-5 tasters. Samples of coffee with antagonists gave tasters taste together with the same sample without antagonists, tasters were asked to identify the more bitter the sample in the pair. As shown in Table 2, tasters each time was determined by the sample fraction coffee without antagonists as more bitter than samples with antagonists, suggesting that antagonists reduced bitter taste in the faction coffee. Also, as shown in Table 3, antagonists reduced the bitter taste of both instant coffee.

As was shown with the help of taste samples from this example, the feeling of bitterness in the compositions (e.g., in food products, beverages and/or drugs), manifested as a bitter taste, can be mitigated or eliminated by making such compositions antagonists hT2R8 and/or hT2R14.

To determine the contribution of individual antagonist conducted taste tests with sredneoblastnymi coffee Connection With. As shown in Table 4, antagonist hT2R14 (Connection C) is sufficient to reduce the bitter taste of coffee in this example.

Example 3

Connection C is an antagonist of the receptor bitter taste a comp the tra actions

The above Example 2 shows that the Compound is an antagonist With T2R person, as determined using high-performance screening analysis using hT2R14. Additional experiments revealed that the antagonist is a wide spectrum of action for 13 T2R person and antagonist narrower spectrum of action for 6 other T2R person. Moreover, in the taste test this compound blocks the intensity of the bitter taste caused near various bitter substances.

In particular, to assess the selectivity of inhibition of Connection, this connection was tested with 22 T2R person, the ligands were determined Senomyx. On these receptors and bitter ligands that activate these T2R man, were described in previous patent applications included in the present description by reference. These 22 T2R are hT2R1, 3, 4, 5, 7, 8, 9, 10, 13, 14, 16, 44, 51, 54, 55, 61, 63, 64, 65, 67, 71 and 75. Amino acid sequence and nucleic acids all of these T2R can be found in earlier patent applications. These T2R person, individually, have to temporarily transfusional in HEK293 cells, stably expressing the promiscuous G-protein G16g44, and with the use of these receptors was carried out functional analyses, as described in those patent applications. In these experiments, each recipe is R activated one of the ligands, selected from the bitter molecules, previously described, to activate specific T2R. The ligands used in the EC80 concentration levels. List of used bitter ligands and concentration of the investigated ligands are given in Table 5 in this example.

Next, to confirm the activity of the compounds in vitro analysis of receptors, the inventors conducted a paired comparison taste tests to determine the effect of compounds in vivo. Tasters were asked to taste the bitter substance with or without a Connection With, and to determine which of the samples is more bitter. Each taster tested several pairs, which resulted in increase of the sample, and the results were analyzed by using appropriate statistical methods. The order of the samples with the Connection With and without him was randomizirana and balanced.

In order to ensure a broad spectrum of antagonistic properties of the compounds, it is tested for the ability to block the bitter taste caused by different bitter ligands, as well as the bitter taste caused bitter ligands whose ability to activate multiple bitter taste receptors are known, and bitter ligands that are not demonstrated the ability to activate specific hT2R. Researched several bitter molecules whose ability to activate the receptor Gorky known that activation of the I which is inhibited by Compound C. In particular, salicin is a bitter molecule that activates hT2R16, and taste test showed that the Connection With the concentration of 40 μm can reduce its bitter taste. Phenyltoloxamine is a bitter molecule that activates hT2R51, and the Connection has reduced its bitter taste at a concentration of 25 ám.

Several bitter molecules able to activate multiple T2R, were similarly tested with the Connection With. the Activation of bitter taste receptors for some of these molecules partially inhibited Connection With. Omeprazole is a bitter molecule that activates hT2R10, 14 and 75. Despite its bitter taste optional mediate multiple receptors bitter, its bitter taste is also significantly reduced the Connection With. Rebaudioside a is a natural sweetener with a strong bitter taste, activating at least 7 T2R person. Its bitter taste is also reduced Connection With.

Additionally, the Connection With inhibited bitter taste for some compounds, the receptors for which are unknown, such as dextromethorphan and diphenylhydramine. Investigated the influence of Connections on these compounds and found that their bitter taste also declined.

In connection with the above data, Figure 5 contains the results of experiments in which the Connection is tested with various compounds, antagony is Tami. Activity inhibition is represented by a decrease in the activity of the receptor in the presence of Compound C. Figure 5 shows that 13 different hT2R significantly (>30%) inhibited by Compound C. These 13 hT2R are hT2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 64, 65 and 71. Six other receptors, including hT2R5, 9, 13, 54, 67 and 75 are also inhibited, but to a lesser extent.

Table 5
The list of ligands and concentrations used for each investigated T2R
hT2RAgonistConcentration
1Picric acid0.05 mm
3Chloroquine pH6,550 µm
4Chloroquine pH6,55 mm
5Picoline10 mm
7Chloroquine pH6,510 mm
8Andrographolide0,5 mm
9Ofloxacin1 mm
10Strychnine50 µm
13Oxepanone1 mm
14Aristolochia acid2 µm
16Salicin1 mm
44Denatonium0.5 µm
51Prop2.5 μm
54Ranitidine5 mm
55Cinchonine150 microns
61Aristolochia acid25 nm
63Caffeine1 mm
63Andrographolide100 mm
64 Aristolochia acid1 micron
65Oleuropein1 mm
67Andrographolide5 µm
71Picric acid10 µm
75Strychnine1 micron

Example 4

Antagonists hT2R8: obtain the compounds according to the invention

Typical compounds according to the invention are synthesized as follows.

Example 4-1: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)picolinate

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (400 mg, 2.1 mmol), Pikalyovo acid (256 mg, 2.1 mmol) and HOBt (388 mg, of 2.50 mmol) were mixed in DHM (7 ml). The reaction mixture was treated with triethylamine (670 ml, 4.8 mmol) and was stirred for 15 minutes at room temperature in a nitrogen atmosphere. Added EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide) (598 mg, 3.1 mmol) and the reaction mixture was stirred for additional 4 hours. Then the reaction mixture was diluted with dichloromethane (5 ml) and washed with aqueous saturated solution is m NaHCO 3(5 ml, 2×) and then aqueous saturated NaCl solution (5 ml). The organic phase was collected, dried and filtered. The solvents were removed under vacuum. The crude product is re-suspended in EtOH (5 ml) and was purified by the method of reversed-phase HPLC (with a gradient of 5%-95% acetonitrile in H2O: 25 minutes). Pure fractions were combined and concentrated to obtain N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)picolinate (372 mg, 60%) as a white solid.1H NMR (CDCl3, 400 MHz): δ of 2.21 (s, 3H), of 2.44 (s, 3H), of 5.05 (s, 2H), 7,49-7,47 (m, 1H), to 7.59 (s, 1H), 7,93-7,88 (dt,J=14, 2 Hz, 1H), 8,07 (s, 1H), 8,24-8,21 (l,J=8 Hz, 1H), 8,61-8,56 (m, 1H), 9,83 (users, 1H). LC/MS; [M+H] calculated for C15H15N5O2; the expected value 297,1; experimentally 298,3. Melting point: 135-137ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.57 μm.

Example 4-1a: 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride

Tert-butyl-1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylcarbamate (example 4-1b) (592 mg, 2 mmol) was stirred in a solution of 4N HCl in dioxane (20 ml) at ambient temperature for 2 hours. The solvent was removed under reduced pressure and the residue was transferred into a mixture of 1/1 ethyl acetate/hexane (30 ml) and concentrated (twice). The solid was ground into powder with hexane and collected by filtration, the floor of the th 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (500 mg, 99%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), of 2.38 (s, 3H), 5,16 (s, 2H), 7,51 (s, 1H), 8,03 (s, 1H), 10,27 (users, 3H).

Example 4-1b: tert-butyl 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylcarbamate

3,5-Dimethyl-4-((4-nitro-1H-pyrazole-1-yl)methyl)isoxazol (example 4-1c) (14.6 g, 66 mmol), Boc anhydride and 10% Pd/C (3.8 g) was stirred in MeOH (400 ml) under 1 atmosphere of H2for 16 hours at ambient temperature. The mixture was filtered and the solution was removed under reduced pressure. The residue was purified by chromatography on silica gel (20% ethyl acetate in hexane) to give tert-butyl 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylcarbamate (12.7 g, 66%) as a light pink solid.1H NMR (CDCl3, 400 MHz): δ of 1.41 (s, 9H), 2,10 (s, 3H), 2,32 (s, 3H), of 4.90 (s, 2H), to 6.19 (users, 1H), 7,19 (s, 1H), 7,50 (s, 1H). MS 293 (MH+).

Example 4-1c: 3,5-dimethyl-4-((4-nitro-1H-pyrazole-1-yl)methyl)isoxazol

To 4-nitro-1H-pyrazole (example 4-1d) (3.8 g, 34 mmol) in DMF (80 ml), cooled to 0ºC with bath ice/water, was addedt-BuOK (4,2 g, 38 mmol). After adding the base bath with ice was removed and the mixture was stirred for 30 minutes followed by the addition of 4-(chloromethyl)for 3,5-dimethylisoxazole (5 g, 34 mmol). The reaction mixture is boiled under reflux for 16 hours, then was cooled to ambient temperature. To the reaction mixture was added a 2O and the precipitate was collected by filtration. The precipitate was washed with additional amount of H2O, then dried in high vacuum to obtain 3,5-dimethyl-4-((4-nitro-1H-pyrazole-1-yl)methyl)isoxazol (5.8 g, 78%) as a pale yellow solid.1H NMR (CDCl3, 400 MHz): δ of 2.23 (s, 3H), of 2.46 (s, 3H), to 5.08 (s, 2H), 8,02 (s, 1H), 8,08 (s, 1H).

Example 4-1d: 4-nitro-1H-pyrazole

Pyrazole (10 g, 147 mmol) was added in portions to concentrated sulfuric acid (100 ml), keeping the temperature of the reaction mixture in the vessel below 50ºC using a bath of ice water. Then was added dropwise concentrated nitric acid (10 ml), maintaining the temperature of the reaction mixture below 50ºC using a bath of ice water. Bath with ice water was removed and the reaction mixture was heated to 60ºC and was stirred for 4 hours. The reaction mixture was cooled with a bath of ice water, and was podslushivaet to ~pH 8 through 18 N aqueous NaOH (150 ml). The product which has precipitated in the form of a white solid substance was collected by filtration, washed with H2O and dried in high vacuum to obtain 4-nitro-1H-pyrazole (7 g, 42%) as a white solid.13C NMR (100 MHz, CDCl3): δ 126,4, 137,0.

Example 4-2: 3-chloro-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-4-(methylsulphonyl)thiophene-2-carboxamide

With stirring to a mixture of ((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (500 mg, 2 mmol) in DHM (20 ml), cooled to 0ºC with a bath of ice water, was added triethylamine (600 mg, 6 mmol). The mixture was stirred until all the solid is not passed into the solution (~10 minutes). 3-Chloro-4-(methylsulphonyl)thiophene-2-carbonyl chloride (543 mg, 2.1 mmol) in 2 ml of CH3CN was added via syringe to the specified free amine at 0ºC. Deleted the bath with ice and the mixture was stirred for 2 hours. The reaction mixture was diluted with dichloromethane (100 ml) and the organic phase is washed with H2O (200 ml). The organic phase was dried over sodium sulfate, filtered and concentrated under reduced pressure. The solid was ground into powder with ethyl acetate/hexane (1/5) and received 3-chloro-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-4-(methylsulphonyl)thiophene-2-carboxamid (375 mg, 45%) as a white solid.1H NMR (CDCl3, 400 MHz): δ of 2.20 (s, 3H), 2,43 (s, 4H), up 3.22 (s, 3H), of 5.05 (s, 2H), EUR 7.57 (s, 1H), 7,94 (s, 1H), to 8.41 (s, 1H), 8,59 (users, 1H). LC/MS; [M+H] 415,5. Melting point: 202-204ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 2,09 microns.

Example 4-3: (S)-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylpropanamide

1-((3,5-Dimethylisoxazol the-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (200 mg, 1 mmol), (S)-2-phenyl propionic acid (156 mg, 1 mmol) and PyBop (650 mg, 1.3 mmol) was added to DMF (4 ml) followed by the addition of triethylamine (0.3 ml, 2.1 mmol). The reaction mixture was stirred for 4 hours at room temperature in a nitrogen atmosphere, and then was diluted with ethyl acetate (20 ml), washed with aqueous saturated solution of NaHCO3(2×15 ml)and then aqueous saturated NaCl solution (15 ml). The organic phase was dried, filtered and concentrated on a rotary evaporator. The crude product is re-suspended in methanol (3 ml) and was purified by the method of reversed-phase HPLC (with a gradient of 5%-95% acetonitrile in H2O: 25 minutes). The fractions containing pure product were concentrated to obtain (S)-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylpropanamide (200 mg, 60%) as a white solid.1H NMR (400 MHz, DMSO-d6) δ 1,36 (l,J=7,2 Hz, 3H), of 2.09 (s, 3H), of 2.36 (s, 3H), 3,71-3,66 (m, 1H), of 5.05 (s, 2H), 7,33-7,17 (m, 5H), 7,37 (s, 1H), of 7.90 (s, 1H), of 10.05 (s, 1H). MS 325 (M+H). Melting point 108ºC - 110ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 were 0.41 μm.

Example 4-4: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(4-hydroxy-3,5-acid)ndimethylacetamide

1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (376 mg, 1.7 mmol), 2-(4-hydro is si-3,5-acid)acetic acid (350 mg, 1.7 mmol), PyBop (1 g, 2 mmol) and triethylamine (605 mg, 6 mmol) were stirred together in DMF (10 ml) at room temperature for 2 hours. The reaction mixture was diluted with aqueous 1N HCl (100 ml) and was extracted with DHM (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by chromatography on silica gel (30% ethyl acetate in hexane) to give N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(4-hydroxy-3,5-acid)ndimethylacetamide (189 mg, 29%) as a white solid.1HNMR (CDCl3, 400 MHz) δ 2,10 (s, 3H), of 2.36 (s, 3H), 3,40 (s, 2H), 3,70 (s, 6H), 5,07 (s, 2H), 6,53 (s, 2H), 7,39 (s, 1H), 7,92 (s, 1H), 8,18 (s, 1H), there is a 10.03 (s, 1H).LC/MS; [M+H] calculated for C19H22N4O5; the expected value 387,16; experimentally 387,6. Melting point: 187-188ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0,46 mm.

Example 4-5: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylpropanamide

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (300 mg, 1.3 mmol), 2-phenylpropane acid (225 mg, 1.5 mmol), triethylamine (300 mg, 3 mmol), DMAP (4-dimethylaminopyridine) (61 mg, 0.5 mmol) and EDC (386 mg, 2 mmol) were stirred together in DHM (10 ml) at room temperature for 4 hours. Reacciona the mixture was diluted aqueous solution of 1N HCl (100 ml) and was extracted with DHM (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by chromatography on silica gel (30% ethyl acetate in hexane) to give N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylpropanamide(272 mg, 81%) as a white solid.1HNMR (CDCl3, 400 MHz) δ of 1.36 (d, 3H, J=7.2 Hz), 2,10 (s, 3H), is 2.37 (s, 3H), 3,70 (m, 1H, J=6,8 Hz), is 5.06 (s, 2H), 7,20 (t, 1H, J=8,4 Hz), 7,31-7,28 (m, 4H), 7,38 (s, 1H), to $ 7.91 (s, 1H), 10,10 (s, 1H).LC/MS; [M+H] calculated for C18H20N4O2; the expected value 325,16; experimentally 325,5. Melting point: 129-130ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was of 0.32 μm.

Example 4-6: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylacetamide

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (230 mg, 1 mmol) and triethylamine (300 mg, 3 mmol) was stirred DHM (10 ml), cooled to 0ºC with bath ice/water. Under stirring to the reaction mixture was added dropwise 2-phenylacetyl chloride (184 mg, 1.3 mmol). When you are finished adding bath with ice was removed and the reaction mixture was stirred for 1 hour. The mixture was diluted DHM (50 ml), washed with 1N aqueous HCl (100 ml)and then 1N aqueous NaOH (100 ml) and then H2O (100 ml). The combined organization is organic extracts were dried over sodium sulfate, was filtered and solvent was removed on a rotary evaporator. The resulting residue was purified by chromatography (50% ethyl acetate in hexane) to give 210 mg of the solid product was ground into powder in ethyl acetate/hexane (1/9) to obtain N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylacetamide (188 mg, 68%) as a white solid.1H NMR (CDCl3, 400 MHz): δ of 2.15 (s, 3H), of 2.38 (s, 3H), of 3.69 (s, 2H), equal to 4.97 (s, 2H), 7,15 (users, 1H), 7,40-7,27 (m, 6H), to 7.84 (s, 1H). LC/MS; [M+H] calculated for C17H18N4O2; the expected value 311,14; experimentally 311,40. Melting point: 106-108ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.53 μm.

Example 4-7: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-methoxybenzamide

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (300 mg, 1.3 mmol), 3-methoxybenzoic acid (172 mg, 1.3 mmol), EDC (386 mg, 2 mmol) and triethylamine (303 mg, 3 mmol) was stirred DHM (5 ml) at ambient temperature for 6 hours. The reaction mixture was diluted DHM (50 ml) and the organic phase is washed with water of 0,1N HCl (150 ml)and then 1N aqueous NaOH (150 ml). The organic phase was dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The crude product was purified by chromatography on Sealy is agile (40% ethyl acetate in hexane) to give 225 mg off-white solid. The solid was ground into powder with ethyl acetate/hexane (1/9) and the white solid was collected by filtration. Pure product was dissolved in absolute ethanol and concentrated on a rotary evaporator (4×, 25 ml) to give N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-methoxybenzamide (185 mg, 43%) as a white solid.1H NMR (CDCl3, 400 MHz): δ of 2.20 (s, 3H), 2,42 (s, 3H), 3,85 (s, 3H), of 5.03 (s, 2H), 7,09-7,06 (m, 1H), 7,37-7,35 (m, 2H), 7,41 (m, 1H), 7,51 (s, 1H), 7,93 (users, 1H), 8,03 (s, 1H). LC/MS; [M+H] calculated for C17H18N4O3; the expected value 327,14; experimentally 327,30. Melting point: 127-129ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.39 μm.

Example 4-8: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzo[d][1,3]dioxol-5-carboxamid

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 1a) (8 mg, 35 mmol) and benzo[d][1,3]dioxol-5-carboxylic acid (7 mg, 42 μmol) each was dissolved in 200 μl of dimethylformamide. Resin-based Si-carbodiimide (70 mg, 70 μmol) were placed in 1.2 ml 96-well plate Greiner, and then added amine and acid. Hydroxybenzotriazole (6 mg, 42 μmol) was dissolved in 100 μl of dimethylformamide and was added into the reaction well. The reaction mixture was shaken overnight at room temperature. Claudine excess carboxylic acid and hydroxybenzotriazole in the reaction mixture was added PS-trisamino resin (35 mg, 70 mmol) and was shaken overnight at room temperature. In the reaction well was added 200 μl of acetonitrile, and shake for 1 minute. The upper clear solution was transferred into a new tablet. The extraction process was repeated more than two times. The solution was evaporated under vacuum to obtain the target product. Yield 6%. MS M+H calculated 341,1, experimentally 341,2.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 amounted to 0.2 μm.

Example 4-9: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dimethoxybenzamide

Was obtained as in example 4-8 of the 2.5-dimethoxybenzoic acid and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a). Exit 13%. MS M+H calculated 357,5 experimentally 357,3.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.17 μm.

Example 4-10: 3-cyano-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzamid

Was obtained as in example 4-8 from 3-cyanobenzoic acid and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a). Output 15%. MS M+H calculated 322,6, experimentally it was 322.3.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 amounted to 0.2 μm.

Example 4-11: N-(1((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-phenylcyclopropanecarboxylic

Was obtained as in example 4-8 1-phenylcyclopropanecarboxylic acid and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a). Yield 6%. MS M+H calculated 337,6 experimentally 337,5.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.25 μm.

Example 4-12: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenylbutane

Was obtained as in example 4-8 from 3-phenylbutanoate acid and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a). Yield 6%. MS M+H calculated 339,6 experimentally 339,5.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.28 μm.

Example 4-13: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1H-pyrrol-2-carboxamide

Was obtained as in example 4-8 from 1H-pyrrole-2-carboxylic acid and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a). Yield 18%. MS M+H calculated 286,6 experimentally 286,3.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.57 μm.

Example 4-14: 2-cyclohexyl-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)ndimethylacetamide

Received in Primera-8 2-cyclohexyloxy acid and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a). Exit 17%. MS M+H calculated 317,6 experimentally 317,4.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.73 ám.

Example 4-15: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)cinnamamide

Was obtained as in example 4-8 from cinnamic acid and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a). Exit 4%. MS M+H calculated 322,6 experimentally 322,4.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.7 μm.

Example 4-16: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)adamantane

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (300 mg, 1.56 mmol), adamantane-1-carboxylic acid (281 mg, 1.56 mmol), PyBop (972 mg, of 1.87 mmol) and triethylamine (438 ml of 3.12 mmol) were mixed in DMF (5 ml). The reaction mixture in an atmosphere of nitrogen was stirred at room temperature for 4 hours. The reaction mixture was diluted with ethyl acetate (4 ml) and washed with saturated solution of NaHCO3(2×, 3 ml) and then saturated NaCl solution (3 ml). The organic phase was extracted, dried and filtered. The solvents were removed under vacuum. The crude product is re-suspended in methanol (4 ml) and purified by HPLC. Pure product was re-dissolved in this is OLE and concentrated under vacuum (3×3 ml) to give N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)adamantane in the form of a white solid in 60% yield. 1H NMR (400 MHz, CDCl3): δ 1,79 is 1.70 (m, 6H), 1.93 and-of 1.92 (m, 6H), 2,08 (users, 3H), of 2.18 (s, 3H), 2,41 (s, 3H), to 4.98 (s, 2H), 7,37 (s, 1H), 7,38 (s, 1H), 7,92 (s, 1H). MS 355 (M+H). Melting point 167-169ºC.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.88 mm.

Additional compounds were synthesized according to similar procedures described in examples 4-1 - 4-16, and experimentally investigated. As was found, these additional compounds have a relatively high level of effectiveness as inhibitors of the bitter taste receptor hT2R8. The results of this study are presented below in table A.

Table A
No. of connectionsConnectionhT2R8 IC50(µm)
4-17
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-4-methoxybenzamide
0,26
4-18
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl) - for 3,5-dimethoxybenzamide
0,28
4-19
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzamide
0,39

4-20
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-4-hydroxy-3-methoxybenzamide
0,48
4-21
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3,4-dimethoxybenzamide
0,56
4-23
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-4-perbenzoic
1,56
4-24
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzamid
2,62
4-25
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,3-dimethoxybenzamide
0,61
4-26
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-(methylsulphonyl)benzamid
0,72

4-33
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(4-methoxyphenyl)ndimethylacetamide
0,99
4-34
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzamide
1,11
4-35
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(2-methoxyphenyl)ndimethylacetamide
1,16
4-36
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzamide
1,85
4-37
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylbutane
0,50

4-38
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzamide
0,53
4-39
2-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylamino)-2-oxo-1-fenilatilamin
0,83
4-40
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-hydroxy-2-phenylpropanamide
0,96
4-41
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzamide
2,10
4-42
(R)-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-phenylpropanamide
3,72

4-43
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxy-2-phenylacetamide
3,43
4-44
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,2-diphenylacetamide
5,19
4-45
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-4-methyl-1H-Pirro is-2-carboxamide
0,72
4-46
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)nicotinamide
1,05
About 4-47
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzamide
1,35
4-48
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-6-methylpyridine
1,95

4-49
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-1H-pyrrol-2-carboxamide
3,40
4-50
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)isonicotinamide
4,59
4-51
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methylfuran-3-carboxamide
7,87
4-52
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3-dimethyl-1H-pyrazole-5-carboxamide
18,05
4-53
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dimethylfuran-3-carboxamide
3,03
4-54
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylfuran-2-carboxamide
3,19

4-55
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenylpropanamide
0,49
4-56
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-4-phenylbutyramide
0,83
4-57
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzamide
4,16
4-58
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(furan-3-yl)ndimethylacetamide
8,66
4-59
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-ethoxybenzene
0,22
4-60
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-methylpentanoic
1,94

4-61
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)cyclohexanecarboxylic
2,20
4-62
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(adamant-1-yl)ndimethylacetamide
of 3.77
4-63
1-((3,5-dimethylisoxazol-4-yl)methyl)-N-methyl-1H-pyrazole-4-amine
8,76
4-64
N,N-bis(2,3-dimethoxybenzyl)-1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine
12,60
4-65
N-(1-((3,5-dimethylisoxazol the-4-yl)methyl)-1H-pyrazole-4-yl)-N-methyl-2-phenylpropanamide
2,81

4-66
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-N-methyl-1-phenylcyclopropanecarboxylic
8,23
4-67
(S)-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxy-2-phenylacetamide
8,6
4-68
(R)-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxy-2-phenylacetamide
5,0
4-69
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(4-hydroxy-3-methoxyphenyl)ndimethylacetamide
the 4.7
4-70
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(3-hydroxy-4-methoxyphenyl)ndimethylacetamide
3,1

4-71
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-hydroxy-3-methoxime Samid
2,9
4-72
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzo[d]oxazol-5-carboxamid
2,6
4-73
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3,4-dihydroxy-5-methoxybenzamide
2,2
4-74
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-methoxy-4-methylbenzamide
2,2
4-75
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methylbenzofuran-5-carboxamid
2,1

4-76
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)cinoxacin-5-carboxamid
1,7
4-77
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzofuran-5-carboxamide
1,4
4-78
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,2-debtorrent[d][1,3]dioxol-5-carboxamid
1,3
4-79
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-1H-benzo[d]imidazol-5-carboxamid
1,0
4-80
3-chloro-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzamid
0,9

4-81
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-ethylbenzo[d]oxazol-5-carboxamid
0,8
4-82
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)cinoxacin-6-carboxamide
0,8
4-83
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methylbenzo[d]oxazol-6-carboxamide
0,7
4-85
N-(1-((3,5-demetrisom the azole-4-yl)methyl)-1H-pyrazole-4-yl)-1H-benzo[d]imidazol-5-carboxamid
0,6
4-86
2-(1H-benzo[d]imidazol-6-yl)-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)ndimethylacetamide
0,6

4-87
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1H-benzo[d][1,2,3]triazole-5-carboxamide
0,6
4-88
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide
0,4
4-89
7-bromo-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide
0,3
4-90
7-chloro-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide
0,1
4-91
8-chloro-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide
0,1

4-96
1-((3,5-dimethylisoxazol-4-yl)methyl)-N-methyl-1H-pyrazole-4-amine
8,764
4-97
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-N-methyl-2-phenylpropanamide
2,126
4-98
(S)-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-N-methyl-2-phenylpropanamide
2,811
4-99
2-cyano-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzosulfimide
1,358
4-100
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-forbindelsesfaneblad
8,510
4-101
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dimethoxybenzenesulfonamide
1,631

4-102

N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methylbenzenesulfonamide
2,153
4-103
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methoxybenzenesulfonamide
3,801
4-104
methyl-3-(N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)sulfamoyl)thiophene-2-carboxylate
1,252
4-105
1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-(2-forfinal)thiourea
1,629
4-106
1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-(2-methoxyphenyl)thiourea
2,607
4-107
1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-(pyridin-3-yl)thiourea
2,999

4-108
1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-o-tolylthiourea
3,013
4-109
1-(3-cyanophenyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)thiourea
0,783
4-110
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)thiourea
1,097
4-111
1-(2-cyanophenyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)thiourea
2,347
4-112
1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenyltoloxamine
2,492
4-113
1-(2,5-acid)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)thiourea
5,240

4-114
N-(2-((3,5-dimethylisoxazol-4-yl)methyl)-2H-tetrazol-5-yl)-3-methoxybenzamide
1,866
4-115
N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-3-methyl-1H-pyrazole-4-yl)-3-methoxybenzamide
9,248
4-116
3-((3,5-dimethylisoxazol-4-yl)methyl)-2-oxo-N-(thiophene-2-ylmethyl)-2,3-dihydrothiazolo-5-carboxamid
2,279

EXAMPLES 4-67 - 4-91:

Was obtained as in example 4-73 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) and the corresponding functionalized carboxylic acids. The characterization was performed by LC/MS, were found the required mass.

Example 4-73: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3,4-dihydroxy-5-methoxybenzamide

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) (228 mg, 1 mmol), 3,4-dihydroxy-5-methoxybenzoic acid (184 mg, 1 mmol), HOBt (135 mg, 1 mmol) and EDC (191 mg, 1 mmol) was dissolved in 2 ml of DMF in a vessel for use in a microwave system, after which was added triethylamine (101 mg, 1 mmol). The reaction mixture was placed in a microwave reactor at 165ºC for 5 minutes. The crude product was purified immediately, using the Varian HPLC (gradient 10%-95% acetonitrile in H2O:25 minutes). Pure fractions were combined and concentrated to obtain N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3,4-dihydroxy-5-methoxybenzamide.(280 mg, 70%). LC/MS; [M+H] calculated for C17H18N4O5; the expected value 359,1; experimentally 359,1.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 2.2 μm.

Example 4-92: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-7-methoxybenzo[d][1,3]dioxol-5-carboxamid

N-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3,4-dihydroxy-5-methoxybenzamide (example 73) (50 mg, 0.14 mmol) and cesium carbonate (113 mg, 2.5 mmol) was dissolved in 1 ml of acetone, and then added dibromoethane (239 mg, 1.4 mmol). The reaction mixture was placed in a microwave reactor at 120ºC for 20 minutes. The obtained transparent solution was removed and evaporated under vacuum. The crude product was dissolved in 1 ml ethanol and purified by HPLC Varian (gradient 10%-95% acetonitrile in H2O: 25 minutes). Pure fractions were combined and concentrated to obtain N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(7-methoxybenzo[d][1,3]dioxol-5-yl)ndimethylacetamide.(12 mg, 23%). LC/MS; [M+H] calculated for C18H18N4O5; the expected value 371,1; experimentally 371,1.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.7 μm.

Example 4-3: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-8-methoxy-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide

Was obtained as in example 4-92 from N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3,4-dihydroxy-5-methoxybenzamide (example 4-73), cesium carbonate and dibromethane. A yield of 20%. MS M+H calculated 385,1, experimentally 385,1.

IC50connection specified in the inhibition of bitter taste receptor hT2R8 was 0.7 μm.

Example 4-94: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-7-methoxy-2-methylbenzo[d][1,3]dioxol-5-carboxamid

Was obtained as in example 4-92 from N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3,4-dihydroxy-5-methoxybenzamide (example 4-73), cesium carbonate and 1,1-dibromoethane. Yield 25%. MS M+H calculated 385,1, experimentally 385,1.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 was 0.7 μm.

Example 4-95: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-7-methoxy-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxamide

Was obtained as in example 4-73 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 4-1a) and 7-methoxy-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxylic acid (example 4-95a). The yield is 50%. MS M+H calculated 385,1, experimentally 385,1.1H NMR (400 MHz, DMSO): δ 2,136 (s, 3H), 2,410 (s, 3H), 3,851 (s, 3H), 4,214 (users, 2H), 4,296 (users, 2H), 5,120 (s, 2H), 6,688 (s, 1H), 7,290 (s, 1H), 7,6006 (s, 1H), 8,069 (s,1H), 9,856 (s, 1H).

IC50connection specified in the inhibition of bitter taste receptor hT2R8 was 0.7 μm.

Example 4-95a: 7-Methoxy-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxylic acid

In a vessel with a capacity of 2 ml for use in the microwave system was dissolved methyl 7-bromo-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxylate (273 mg, 1 mmol) and CuBr (14.3 mg, 0.1 mmol) in dried DMF and placed in an ice bath. (540 mg, 10 mmol) In the reaction mixture under stirring was added dropwise sodium methoxide at 0ºC. The reaction mixture was heated to room temperature and was stirred for 45 minutes. Then the reaction mixture was placed in a microwave reactor for 5 min at 135ºC. The reaction mixture was dissolved in water and washed with ethyl acetate. The aqueous phase was collected and acidified to pH 4 with 1M HCl. The product was extracted using ethyl acetate, then dried over sodium sulfate. The solvent was evaporated under vacuum to obtain the desired intermediate compounds, representing 7-methoxy-2,3-dihydrobenzo[b][1,4]dioxin-6-carboxylic acid which was used directly without further purification. Yield 57%. MS M+H calculated 211,1, experimentally 211,1.

Example 5 Antagonists hT2R14: obtain the compounds according to the invention

The following examples are given to illustrate the I various typical variants of realization of the invention and in no way limit the present invention.

Example 5-1: 4-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Benzyl-4-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-1a) (517 mg, 1 mmol) was stirred solution 10/1/2 6N NaOH(aqueous)/THF/MeOH (27 ml) at ambient temperature for 6 hours. The solution was acidified 3N HCl (aqueous) to pH ~3 (about 50 ml) and the aqueous phase was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate and concentrated on a rotary evaporator. The residue was transferred in MeOH (15 ml) and was purified by the method of reversed-phase HPLC (with a gradient of 5-95% acetonitrile in H2O: 25 minutes), receiving three aliquots of 5 ml Pure fractions were combined and concentrated to a white solid. The product was dissolved in 15 ml of absolute ethanol and evaporated on a rotary evaporator (4×) to give the pure 4-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)benzoic acid (174 mg, 42%) as a white solid. TPL 161-163ºC.1H NMR (CDCl3, 400 MHz): δ of 3.78 (s, 3H), 4,32 (s, 2H), 4,36 (s, 2H), 6,78 (l,J=8,4 Hz, 2H), 6,99 (l,J=8,8 Hz, 2H), 7,09-7,07 (m, 2H), 7,27-7,24 (m, 3H), of 7.93 (d,J=8,8 Hz, 2H), 8,24 (e,J=8 Hz, 2H).MS 412(MH+).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 amounted to 0.22 μm.

Example 5-1a: Benzyl 4-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)benzo is t

4-(N-(4-methoxybenzyl)sulfamoyl)benzoic acid (example 5-1b) (450 mg, 1.4 mmol), benzylbromide (770 mg, 4.5 mmol) and cesium carbonate (1.5 g, 4.5 mmol) in DMF (10 ml) was stirred at 80ºC for 2 hours. The solution was cooled to room temperature, diluted with H2O (200 ml) and was extracted with ethyl acetate (3×, 100 ml). The combined organic phases were dried over sodium sulfate, filtered and concentrated under vacuum. The residue was purified by chromatography on silica gel (10% ethyl acetate in hexane) to give benzyl-4-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)benzoate (517 mg, 73%) as a white solid.1H NMR (400 MHz, CDCl3)δ3,76 (s, 3H), 4,28 (s, 2H), 4,32 (s, 2H), of 5.40 (s, 2H), 6,79 (l,J=8 Hz, 2H), of 6.96 (d,J=8 Hz, 2H),? 7.04 baby mortality-7,07 (m, 2H), 7,21-of 7.23 (m, 3H), 7,35-7,47 (m, 5H), the 7.85 (d,J=8 Hz, 2H), 8,17 (l,J=8,4 Hz, 2H).

Example 5-1b: 4-(N-(4-methoxybenzyl)sulfamoyl)benzoic acid

4-(Chlorosulfonyl)benzoic acid (5 g, 22.7 mmol) as a solid substance was added in three portions with stirring to a solution of 4-methoxybenzylamine (6,1 g, 45 mmol) and triethylamine (2.3 g, 22.7 mmol) in acetone (100 ml), cooled to 0ºC with a bath of ice water for 10 minutes. Bath ice was removed and the reaction mixture was stirred for additional 4 hours. The reaction mixture was diluted with a solution of 5% acetic acid in H2O (150 ml) and was extracted with ethylacetate the (3×, 100 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The resulting white solid was ground into powder with hexane/ethyl acetate (9/1) to obtain 4-(N-(4-methoxybenzyl)sulfamoyl)benzoic acid (5.1 g, 70%) as a white solid.1H NMR (400MHz, DMSO-d6) δ 3,68 (s, 3H), 3,91 (s, 2H), 6,79 (l,J=8,4 Hz, 2H), 7,10 (l,J=8,8 Hz, 2H), 7,80 (l,J=8,8 Hz, 2H), 8,02 (l,J=8,4 Hz, 2H).

Example 5-2: 4-(N-(furan-2-ylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

4-Methoxybenzyl-4-(N-(furan-2-ylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-2a) (750 mg, 1.4 mmol) was stirred in a mixture 2/2/1 2N aqueous LiOH/THF/MeOH (45 ml) at ambient temperature for 3 hours. The solution was acidified using 1N aqueous HCl to pH ~3 (about 100 ml) and was extracted with ethyl acetate (3×, 100 ml). The combined organic extracts were dried over sodium sulfate and concentrated on a rotary evaporator. The residue was transferred in MeOH (9 ml) and was purified by the method of reversed-phase HPLC (with a gradient of 5-95% acetonitrile in H2O: 40 minutes), receiving three aliquots of 3 ml Pure fractions were combined and concentrated to a white solid. The product was dissolved in absolute ethanol and evaporated (4×20 ml) to obtain pure 4-(N-(furan-2-ylmethyl)-N-(4-shall ethoxybenzyl)sulfamoyl)benzoic acid (205 mg, 36%) as a white solid. TPL 151-152ºC.1H NMR (DMSO-d6, 400 MHz): δ 3,71 (s, 3H), 4,24 (s, 2H), 4,27 (s, 2H), 6,14 (d, 1H, J=3.2 Hz), of 6.26 (m, 1H), 6.87 in (d, 2H, J=9,2 Hz), 7,14 (d, 2H, J=8,8 Hz), 7,41 (s, 1H), 7,89 (d, 2H, J=8 Hz), of 8.06 (d, 2H, J=8,4 Hz), 13,48 (users, 1H). MS 400 (M-H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0.59 μm.

Example 5-2a: 4-methoxybenzyl 4-(N-(furan-2-ylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoate:

4-(N-(furan-2-ylmethyl)sulfamoyl)benzoic acid (example 5-2b) (500 mg, 1.8 mmol), p-methoxybenzylamine (624 mg, 4.0 mmol) and cesium carbonate (1.3 g, 4.0 mmol) was dissolved in DMF (10 ml) and stirred at 80ºC for 1 hour. The mixture was cooled to ambient temperature, diluted with H2O (200 ml) and was extracted with ethyl acetate (3×100 ml). The combined organic substances were dried over sodium sulfate and concentrated on a rotary evaporator. The product was purified by chromatography on silica gel (15% ethyl acetate in hexane) to give 4-methoxybenzyl-4-(N-(furan-2-ylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoate (753 mg, 80%) as a clear oil.1H NMR (DMSO-d6, 400 MHz): δ 3,70 (s, 3H), of 3.75 (s, 3H), 4,22 (s, 2H), 4.26 deaths (s, 2H), of 5.39 (s, 2H), 6,14 (d, 1H, J=3.2 Hz), and 6.25 (m, 1H), 6.87 in (d, 2H, J=8,8 Hz), 6,97 (d, 2H, J=8,8 Hz), 7,13 (d, 2H, J=8,4 Hz), 7,39 (m, 1H), the 7.43 (d, 2H, J=8,4 Hz), to $ 7.91 (d, 2H, J=8,4 Hz), 8,07 (d, 2H, J=8,4 Hz).

Example 5-2b: 4-(N-(furan-2-ylmethyl)sulfamoyl)benzoic acid:

4-(Chlorosulfonyl)benzoic acid (5.0 g, 22.7 mmol) under stirring was added in three portions over 10 minutes to a solution of furfurylamine (6.6 g, 68 mmol) in acetone (200 ml), cooled to 0ºC with a bath of ice water. After adding sulphonylchloride bath with ice was removed and the solution was stirred for 1 hour at ambient temperature. The mixture was concentrated and treated by chromatography on silica gel (90% ethyl acetate, 8% hexane and 2% acetic acid) to obtain 4.4 g of 4-(N-(furan-2-ylmethyl)sulfamoyl)benzoic acid (4.4 g, 68%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ Android 4.04 (d, 2H, J=6 Hz), 6,13 (d, 1H, J=3.2 Hz), and 6.25 (m, 1H), 7,43 (m, 1H), 7,83 (d, 2H, J=8,4 Hz), with 8.05 (d, 2H, J=8,4 Hz), at 8.36 (t, 1H, J=6 Hz), 13,4 (users, 1H).

Example 5-3: 4-(N-(4-ethoxybenzyl)-N-(furan-2-ylmethyl)sulfamoyl)benzoic acid

4-Cyano-N-(4-ethoxybenzyl)-N-(furan-2-ylmethyl)benzosulfimide (example 5-3a) (300 mg, 0.8 mmol) were mixed in a 1/1 mixture of dioxane/a 1.5 N aqueous NaOH (100 ml) at 80ºC for 16 hours. The mixture was cooled, acidified using 1N aqueous HCl (100 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The solid was ground into powder with ethyl acetate/hexane (~1/9) and was collected using a filter, the Finance obtaining 4-(N-(4-ethoxybenzyl)-N-(furan-2-ylmethyl)sulfamoyl)benzoic acid (250 mg, 69%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ of 1.29 (t,J=6,8 Hz, 3H), 3,97 (kV,J=6,4 Hz, 2H), 4,23 (s, 2H), 4,27 (s, 2H), 6,15 (l,J=3.2 Hz, 1H), 6,27 (m, 1H), 6,84 (l,J=8,8 Hz, 2H), 7,11 (l,J=8,8 Hz, 2H), 7,39 (m, 1H), 7,87 (l,J=8,4 Hz, 2H), 8,00 (l,J=8,4 Hz, 2H).

IC50connection with the inhibition of bitter taste receptor hT2R14 was 3.0 μm.

Example 5-3a: 4-cyano-N-(4-ethoxybenzyl)-N-(furan-2-ylmethyl)benzosulfimide:

4-Cyanobenzoyl-1-sulphonylchloride (600 mg, 2.9 mmol) was added under stirring to a solution of N-(4-ethoxybenzyl)-1-(furan-2-yl)methanamine (example 5-3b) (685 mg, 2.9 mmol) and triethylamine (455 mg, 4.5 mmol) in DHM (100 ml) and the reaction mixture was stirred for 2 hours. The reaction mixture was diluted with H2O (200 ml) and was extracted with DHM (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by chromatography on silica gel (10% ethyl acetate in hexane) to give 4-cyano-N-(4-ethoxybenzyl)-N-(furan-2-ylmethyl)benzosulfimide (665 mg, 68%) as off-white solid.1H NMR (DMSO-d6, 400 MHz): δ of 1.29 (t,J=7.2 Hz, 3H), 3,97 (kV,J=6,8 Hz, 2H), 4,23 (s, 2H), 4,27 (s, 2H), 6,15 (l,J=3.2 Hz, 1H), 6,27 (m, 1H), 6,84 (l,J=8,8 Hz, 2H), 7,11 (l,J=8,8 Hz, 2H), 7,39 (m, 1H), 7,92 (l,J=8,4 Hz, 2H), 8,03 (l,J=8,4 Hz, 2H).

Example 5-3b: 4 N-(4-ethoxybenzyl)-1-(furan-2-yl)meta is Amin:

4-Ethoxybenzaldehyde (5 g, 33 mmol) and furfurylamine (4,2 g, 43 mmol) in a mixture of methanol (50 ml), triethylorthoformate (10 ml) and AcOH (1 ml) was stirred at room temperature under nitrogen atmosphere for 16 hours. Was added sodium borohydride (1.4 g, 35 mmol) in 4 portions over 30 minutes (exothermic reaction). The reaction mixture was stirred for additional 2 hours at room temperature. The solvent was removed under vacuum and the residue was transferred into ethyl acetate (150 ml). The organic phase is washed with H2O (200 ml) and the aqueous phase was again extracted with ethyl acetate (2×, 100 ml). The combined organic phases were concentrated and the residue was purified on silica gel (70% ethyl acetate in hexane with ~0.5% triethylamine) to give N-(4-ethoxybenzyl)-1-(furan-2-yl)methanamine (6,1 g, 80%) as a clear oil.1H NMR (CDCl3, 400 MHz): δ 1,40 (t,J=7.2 Hz, 3H), 3,71 (s, 2H), 3,76 (s, 2H), was 4.02 (kV,J=7.2 Hz, 2H), 6,17 (l,J=4 Hz, 1H), of 6.31 (m, 1H), 6,84 (l,J=8,8 Hz, 2H), 7.23 percent (l,J=8,8 Hz, 2H), was 7.36 (m, 1H).

Example 5-4: 4-(N-ethyl-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

4-(N-(4-methoxybenzyl)sulfamoyl)benzoic acid (example 5-1b) (160 mg, 0.5 mmol) and cesium carbonate (325 mg, 1 mmol) was placed in a vessel for use in the microwave system and was dissolved in 2 ml of DMF. In the reaction mixture was added ethyliodide (155 mg, 1 mmol). The reaction to the offer was placed in a microwave reactor and heated at 165ºC for 5 minutes. The reaction mixture was dissolved in ethyl acetate and washed with water. The organic phase was dried over sodium sulfate and evaporated in vacuum. The crude product was dissolved in a solution 4/1 6N NaOH (aqueous)/tetrahydrofuran (3 ml) and stirred at ambient temperature for 6 hours. The solution was acidified 3N HCl (aqueous) to pH ~3 and the product was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate and concentrated under vacuum. The residue was transferred to a methanol (3 ml) and was purified by the method of reversed-phase HPLC (with a gradient of 5-95% acetonitrile in H2O: 25 minutes). As is known, these compounds inhibit hT2R14 with IC5020 microns. Yield 35%. MS M+H calculated 350,11 experimentally 350,0.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 10 μm.

Example 5-5: 4-(N-benzyl-N-(furan-2-ylmethyl)sulfamoyl)benzoic acid

4-(N-(furan-2-ylmethyl)sulfamoyl)benzoic acid (example 5-2b) (140 mg, 0.5 mmol) and cesium carbonate (325 mg, 1 mmol) was placed in a vessel for use in the microwave system and was dissolved in 2 ml of DMF. In the reaction mixture was added methyl bromide)benzene (170 mg, 1 mmol). The reaction mixture was placed in a microwave reactor and heated at 165ºC for 5 minutes. The reaction mixture was dissolved in ethyl acetate and washed the water. The organic phase was dried over sodium sulfate and evaporated in vacuum. The crude product was dissolved in a 4/1 solution of 6N NaOH (aqueous)/tetrahydrofuran (3 ml) and stirred at ambient temperature for 6 hours. The solution was acidified 3N HCl (aqueous) to pH ~3 and the product was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate and concentrated in vacuum. The residue was transferred to a methanol (3 ml) and was purified by the method of reversed-phase HPLC (with a gradient of 5-95% acetonitrile in H2O: 25 minutes). Yield 35%. MS M+H calculated 372,4 experimentally 372,0.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 4.6 μm.

Example 5-6: 4-(N-(furan-2-ylmethyl)-N-(3-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-5 of 1-(methyl bromide)-3-methoxybenzene and 4-(N-(furan-2-ylmethyl)sulfamoyl)benzoic acid (example 5-2b). Yield 35%. MS M+H calculated 402,3 experimentally 402,0.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 10 μm.

Example 5-7: 4-(N-(furan-2-ylmethyl)-N-(2-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-5 of 1-(methyl bromide)-2-methoxybenzene and 4-(N-(furan-2-ylmethyl)sulfamoyl)benzoic acid (when is EP 5-2b). Yield 35%. MS M+H calculated 402,3 experimentally 402,0.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 12 μm.

Example 5-8: 4-(N-(4-propoxyphenyl)-N-(furan-2-ylmethyl)sulfamoyl)benzoic acid

4-Cyano-N-(4-propoxyphenyl)-N-(furan-2-ylmethyl)benzosulfimide (example 5-8a) (300 mg, 0.8 mmol) were mixed in a 1/1 mixture of dioxane/a 1.5 N aqueous NaOH (100 ml) at 80ºC for 16 hours. The mixture was cooled, acidified using 1N aqueous HCl (100 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The solid was ground into powder with ethyl acetate/hexane (~1/9) and collected by filtration to obtain 4-(N-(4-propoxyphenyl)-N-(furan-2-ylmethyl)sulfamoyl)benzoic acid (165 mg, 63%) as a white solid.1HNMR (DMSO-d6, 400 MHz): δ 0,94 (t,J=7,6 Hz, 3H), 1.70 to (m,J=6,8 Hz, 2H), a 3.87 (t,J=6,4 Hz, 2H), 4,23 (s, 2H), 4,27 (s, 2H), 6,13 (l,J=2,8 Hz, 1H), 6,27 (m, 1H), 6,84 (l,J=6,8 Hz, 2H), 7,11 (l,J=8,8 Hz, 2H), 7,39 (m, 1H), 7,87 (l,J=6,8 Hz, 2H), with 8.05 (d,J=6,8 Hz, 2H), 13,45 (users, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 2.5 μm.

Example 5-8a: 4-cyano-N-(4-propoxyphenyl)-N-(furan-2-ylmethyl)benzosulfimide

4-Qi is nebenzal-1-sulphonylchloride (600 mg, 2.9 mmol) was added under stirring to a solution of N-(4-propoxyphenyl)-1-(furan-2-yl)methanamine (example 5-8b) (685 mg, 2.9 mmol) and triethylamine (455 mg, 4.5 mmol) in DHM (100 ml) and the reaction mixture was stirred for 2 hours. The reaction mixture was diluted with H2O (200 ml) and was extracted with DHM (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by chromatography on silica gel (10% ethyl acetate in hexane) to give 4-cyano-N-(4-propoxyphenyl)-N-(furan-2-ylmethyl)benzosulfimide (500 mg, 50%) as off-white solid.1HNMR (DMSO-d6, 400 MHz): δ of 0.95 (t,J=7.2 Hz, 3H), 1.70 to (m,J=6,4 Hz, 2H), 3,88 (t,J=6,4 Hz, 2H), 4,25 (s, 2H), 4,28 (s, 2H), 6,15 (l,J=3.2 Hz, 1H), 6,27 (m, 1H), 6,84 (l,J=6,8 Hz, 2H), 7,11 (l,J=8,8 Hz, 2H), 7,39 (m, 1H), to 7.93 (d,J=6,4 Hz, 2H), 8,01 (l,J=6,4 Hz, 2H).

Example 5-8b: 4 N-(4-propoxyphenyl)-1-(furan-2-yl)methanamine:

4-Propoxybenzaldehyde (5 g, 31 mmol) and furfurylamine (3.9 g, 40 mmol) in a mixture of methanol (50 ml), triethylorthoformate (10 ml) and AcOH (~1 ml) was stirred at room temperature under nitrogen atmosphere for 16 hours. Was added in 4 portions sodium borohydride (1.4 g, 35 mmol) for 30 minutes (exothermic reaction). The reaction mixture was stirred for additional 2 hours at room temperature. The solvent UD is ranged on a rotary evaporator, and the residue was transferred into ethyl acetate (150 ml). The organic phase is washed with H2O (200 ml) and the aqueous phase was again extracted with ethyl acetate (2×, 100 ml). The combined organic phases were concentrated and the residue was purified on silica gel (70% ethyl acetate in hexane with ~2% triethylamine) to give N-(4-propoxyphenyl)-1-(furan-2-yl)methanamine (5,3 g, 75%) as a yellow oil.1H NMR (CDCl3, 400 MHz): δ of 1.03 (t,J=7.2 Hz, 3H), 1,79 (m,J=6,4 Hz, 2H), 3,71 (s, 2H), 3,76 (s, 2H), 3,90 (t,J=6,8 Hz, 2H), 6,17 (l,J=3.2 Hz, 1H), 6,32 (m, 1H), 6,85 (l,J=8,4 Hz, 2H), 7,22 (l,J=8,8 Hz, 2H), 7,37 (m, 1H).

Additional connections are experimentally investigated and found that they have a relatively high level of effectiveness as inhibitors of the bitter taste receptor hT2R14. The results of this study are presented below in table B.

Table B
No. of connectionsConnectionhT2R14 IC50(µm)
5-9
4-(N,N-diisobutylamine)benzoic acid
15

Example 5-10: 4-(N-(4-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Methyl-4-(N-(4-f is orbenin)-N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10a) (3.7 g, 8.3 mmol) was dissolved in MeOH/THF (1:1,5, 30 ml) and was treated with aqueous NaOH (3 N, 15 ml). The mixture was stirred at ambient temperature overnight, then MeOH and THF were removed in vacuo. The resulting aqueous solution was acidified using 6 N aqueous HCl to pH ~3 and was extracted with EtOAc (3×40 ml). The combined organic phases are washed with water and with brine, dried over Na2SO4and concentrated. The crude product was purified by recrystallization from EtOH to obtain pure 4-(N-(4-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid as a white crystalline solid (2.1 g, 58.6 per cent). MS (M-H, 428,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,66 (s, 3H), 4,23 (s, 2H), 4.26 deaths (s, 2H), 6,72 (d, 2H, J=8 Hz), to 6.95 (m, 6H), 7,92 (d, 2H, J=8 Hz), 8,07 (d, 2H, J=8 Hz). IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 1,97 microns.

Example 5-10a: Methyl-4-(N-(4-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoate

Methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) (4.3 g, 12.8 mmol) was dissolved in acetone (70 ml). Added cesium carbonate (to 8.57 g, 25.6 mmol) and 4-florantyrone (1,76 ml, 14,08 mmol) and stirred the mixture at room temperature over night. Inorganic salts were filtered off and the acetone was removed in vacuum.The residue was re-dissolved in ethyl acetate, washed with water and with brine, then the organic phase is sewed over magnesium sulfate and concentrated. The crude product was purified by recrystallization from ethyl acetate/hexane to obtain pure methyl-4-(N-(4-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoate (3.7 g, 65%) as a white solid. 1H NMR (400 MHz, CDCl3): δ million parts of 3.77 (s, 3H), 3,98 (s, 3H), 4,27 (s, 2H), 4,28 (s, 2H), 6,74 (d, 2H, J=8 Hz), 6,92 (m, 4H), 7,03 (m, 2H), 7,88 (d, 2H, J=8 Hz), 8,16 (d, 2H, J=8 Hz).

Example 5-10b: Methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate

To a solution of methyl-4-(chlorosulfonyl)benzoate (example 5-10c) (4 g, 17,09 mmol) in dichloromethane (40 ml) at 0ºC in an ice bath was added (4-methoxyphenyl)methanamine (2,56 ml, 19,65 mmol) and triethylamine (2.38 ml of 17.1 mmol). Then bath with ice was removed and leave the mixture to warm to ambient temperature with stirring for an additional 2 hours. After completion of the reaction (monitored by TLC, 40% ethyl acetate/hexane), the solvent was removed in vacuum. The residue was re-dissolved in ethyl acetate (200 ml), washed with 1N HCl (aqueous, 20 ml), water (20 ml) and with brine (20 ml), then dried over magnesium sulfate. The solution was concentrated and the product was purified by means of recrystallization from hot ethyl acetate/hexane to obtain pure methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (4.3 g, 74,8%) as a white solid. 1H NMR (400 MHz, DMSO-d6): δ (million parts: to 3.67 (s, 3H), of 3.78 (s, 3H), 3,93 (s, 2H), 6,78 (m, 2H), to 7.09 (m, 2H), 7,87 (m, 2H), 8,08 (m, 2H), 8,25 (ears is .c, 1H).

Example 5-10c: Methyl-4-(chlorosulfonyl)benzoate

4-Chlorosulfonylbenzoic acid (5 g, 23 mmol) and thionyl chloride (20 ml) in dichloroethane (10 ml) was heated to 80ºC for 2 hours. The reaction mixture was concentrated by rotary evaporation to obtain a brownish solid. The solid was cooled on ice for 5 minutes and with stirring was added ice-cold methanol (40 ml) at 0ºC for 5 minutes. The reaction mixture was left to warm to ambient temperature and was stirred an additional 10 minutes After adding ice water (40 ml) was obtained white solid, which was collected by filtration and dried under vacuum to obtain pure methyl-4-(chlorosulfonyl)benzoate (4.5 g, 84%). 1H NMR (400 MHz, DMSO-d6): δ (million parts of 3.84 (s, 3H), of 7.70 (d, 2H, J=8,4 Hz), to 7.93 (d, 2H, J=8,4 Hz).

Example 5-11: 4-((N-benzyl-4-methylphenylsulfonyl)methyl)cyclohexanecarbonyl acid

To a suspension of 4-(aminomethyl)cyclohexanecarboxylic acid (of 1.57 g, 10 mmol) in 100 ml of 2,2-dimethoxypropane was added HCl (10 ml, 36% water). The mixture was stirred at ambient temperature for 18 h and then concentrated. The residue was dissolved in minimum amount of MeOH was added diethyl ether to precipitate the salt of HCl, methyl-4-(aminomethyl)cyclohexanecarboxylate as off-white solid in the society. This substance was used without further purification or characterization.

To a mixture of the HCl salt of methyl-4-(aminomethyl)cyclohexanecarboxylate (208 mg, 1 mmol) in 5 ml of dichloromethane at 0ºC in an ice bath was added triethylamine (360 μl, 2.58 mmol) and 4-methylbenzol-1-sulphonylchloride (190 mg, 1 mmol). The mixture in a bath of ice was left to slowly warm to ambient temperature and was stirred overnight. The solvent was removed in vacuum. The residue was re-dissolved in ethyl acetate (20 ml), washed with 1N HCl (5 ml), water (5 ml) and with brine (5 ml), then dried over magnesium sulfate and concentrated. The crude product (162 mg, 0.5 mmol) was re-dissolved in acetone (5 ml) and treated with potassium carbonate (110 mg, 0.79, which mmol) and (4-forfinal)methanamine (1,76 ml, 14,08 mmol). The mixture was stirred in a pressure vessel at 80ºC overnight, then cooled and filtered inorganic salts. The acetone was removed under vacuum, and the residue was re-dissolved in ethyl acetate and washed with water and then with brine. The organic phase was dried over magnesium sulfate and concentrated. The crude product (162 mg, 0.4 mmol) was dissolved in MeOH/THF (1:1,5, 10 ml) and was treated with aqueous NaOH (10N, 400 μl). The mixture was stirred at 100ºC for 20 min in a microwave oven and then remove MeOH and THF in vacuo. The residue was acidified using 6 N aqueous HCl to pH ~3 and what was xtraceroute EtOAc; the combined organic phases are washed with water and with brine, dried over Na2SO4and concentrated. The crude product was purified by recrystallization from EtOH to obtain pure 4-((N-benzyl-4-methylphenylsulfonyl)methyl)cyclohexanecarboxylic acid as a white solid (120 mg, 74%). MS (M+H, 402); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 0.68 (m, 2H), of 0.91 (m, 2H), 1.06 a (users, 1H), 1,50 (d, 2H), 1,72 (d, 2H), 2,0 (1H), is 2.41 (s, 3H), of 2.86 (m, 2H), is 4.21 (s, 2H), 7,30 (m, 5H), the 7.43 (m, 2H), 7,73 (m, 2H), of $ 11.97 (users, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 constituted 0.014 microns.

Example 5-12: 4-(N-(3-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) (500 mg, 1,49 mmol), 3-florantyrone (280 mg, 2,98 mmol) and cesium carbonate (971 mg, 2,98 mmol) were placed in DMF (12 ml) and stirred at 90ºC for 4 hours. The solution was cooled to ambient temperature, diluted with H2O (200 ml) and was extracted with ethyl acetate (3×, 100 ml). The combined organic phases were dried over MgSO4, filtered and concentrated in vacuum. The residue was purified by chromatography on silica gel (10-20% ethyl acetate in hexane) to obtain methyl 4-(N-(3-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoate (528 mg, 80%) as a white solid washes the VA.

Methyl-4-(N-(3-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoate (500 mg, 1.12 mmol) was dissolved in MeOH/THF (1:1, 40 ml) and treated with a solution of aqueous NaOH (10 N, 8 ml). The mixture was stirred at ambient temperature overnight, then remove MeOH and THF by rotary evaporation. The resulting aqueous solution was washed EtOAc (10 ml) and acidified using 6 N aqueous HCl (~15 ml) to pH~4. The aqueous solution was extracted with EtOAc (3×40 ml) and the combined organic phases are washed with water, with brine, dried over MgSO4and concentrated. The crude product was purified by recrystallization from EtOH to obtain the title compound 4-(N-(3-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid as a white crystalline solid (150 mg) with 30% output.

MS (M-H, 428,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts: the 3.65 (s, 3H), 4,27 (s, 2H), 4,29 (s, 2H), 's 6.75 to 7.00 (m, 7H), 7,20 (m, 1H), 7,95 (d, 2H, J=8 Hz); 8,10 (d, 2H, J=8 Hz).

Example 5-13: 4-(N-benzyl-N-(2,4-dimethoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from (2,4-acid)methanamine, methyl-4-(chlorosulfonyl)benzoate (example 5-10c) and benzylbromide. MS (M-H, 440,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,50 (s, 63H), 3,66 (s, 3H), 4,20 (s, 2H), 4,34 (s, 2H), 6,29 (s, 1H), 6,33 (d, J=8.0 Hz, 1H), 6,91 (d, J=8,8 Hz, 1H), 7,12-of 7.23 (m, 5H), 7,80 (d, J=8,0 Hz, 2H), 8,01 (d, J=8,4 Hz, 2H), 13,49 (s, 1H).

Example 5-14: 4-(N-(3-Chlorobenzyl)-N-(4-label benzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and 1-(methyl bromide)-3-chlorobenzene. MS (M-H, 444,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 3.69 (s, 3H), 4,30 (m, 4H), 6,76-7,24 (m, 8H), to 7.99 (m, 2H), 8,13 (m, 2H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 1.88 microns.

Example 5-15: 4-(N-benzyl-N-(2,4,6-trimethoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from (2,4,6-trimethoxyphenyl)methanamine, methyl-4-(chlorosulfonyl)benzoate (example 5-10c) and benzylbromide. MS (M-H, 470,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 3.45 (s, 6H), of 3.69 (s, 3H), 4.26 deaths (s, 2H), 4,28 (s, 2H), 5,98 (s, 2H), 7,11-7,26 (m, 5H), of 7.82 (d, J=8.0 Hz, 2H), 8,07 (d, J=8 Hz, 2H), 13,49 (s, 1H). Elemental analysis (experimentally, %): C 61,05; N 5,49; N 2,98; (calculated %): C 61,13; N 5,34 and N 2,97.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 10.76 microns.

Example 5-16: 4-((N-benzyl-N-(4-methoxybenzyl)sulfamoyl)methyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and benzylbromide. MS (M-H, 424,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts: and 3.72 (s, 3H), 4,18 (d, 2H), 4,55 (s, 2H), 6,83 (m, 2H), 7,12 (m, 2H), 7,21 (m, 2H), 7,28 (m, 3H), 7,43 (m, 2H), to 7.93 (m, 2H).

Example 5-17: 4-(N-(4-methoxybenzyl)-N-propylsulfonyl)is entina acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and n-propyl bromide. MS (M-H, 362,1); 1H NMR (400 MHz, CDCl3): δ million parts: 0,70 (m, 3H), of 1.35 (m, 3H), is 3.08 (m, 2H), of 3.73 (s, 3H), or 4.31 (s, 2H), 6,83 (m, 2H), 7,17 (m, 2H), to 7.93 (m, 2H), 8,23 (m, 2H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 amounted to 3.75 microns.

Example 5-18: 4-(N-(4-methoxybenzyl)-N-phenylsulfonyl)benzoic acid

Was obtained as in example 5-10 from aniline, methyl-4-(chlorosulfonyl)benzoate (example 5-10c) and 1-(chloromethyl)-4-methoxybenzene. MS (M-H, 396,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,66 (s, 3H), to 4.73 (s, 2H), 6,78 (d, J=8,4 Hz, 2H), 7,00 (d, J=7,6 Hz, 2H), 7,12 (d, J=8.0 Hz, 2H), 7,24 (d, J=8.0 Hz, 2H), 8,11 (d, J=8,4 Hz, 2H), 13,49 (s, 1H).

Example 5-19: 4-(N-(4-methoxybenzyl)-N-genetically)benzoic acid

Was obtained as in example 5-10 from 2-fenilalanina, methyl-4-(chlorosulfonyl)benzoate (example 5-10c) and 1-(chloromethyl)-4-methoxybenzene. MS (M-H, 424,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 2.51 (m, 2H), 3,23 (m, 2H), 3,74 (s, 3H), or 4.31 (s, 2H), or 4.31 (s, 2H), 6,92 (m, 2H), 6,98 (m, 2H), 7,20-7,27 (m, 5H), the 7.85 (m, 2H), of 8.06 (m, 2H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 4,58 mm.

Example 5-20: 4-((N-(4-terbisil)-4-methylphenylsulfonyl)methyl)cyclohexanecarbonyl acid/p>

Was obtained as in example 5-11 of 1-(methyl bromide)-4-fervently, 4-(aminomethyl)cyclohexanecarboxylic acid and 4-methylbenzol-1-sulphonylchloride. MS (M+H, 420); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 0.71 (m, 2H), 0,95 (m, 2H), 1,15 (users, 1H)and 1.51 (d, 2H), 1,76 (d, 2H), 2,0 (1H), 2.40 a (s, 3H), 2,85 (m, 2H), 4,22 (s, 2H), 7,14 (m, 2H), 7,32 (m, 2H), 7,40 (m, 2H), 7,69 (m, 2H), 11,93 (users, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0,083 mm.

Example 5-21: 4-((4-acetamido-N-benzilpenitsillino)methyl)cyclohexanecarbonyl acid

Was obtained as in example 5-11 from acetamidobenzoyl-1-sulphonylchloride, 4-(aminomethyl)cyclohexanecarboxylic acid and benzylbromide. MS (M+H 445,2); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 0.65 (m, 2H), of 0.68 (m, 2H), of 1.05 (m, 1H), 1,48 (m, 2H), 1.70 to (m, 2H), 1,92 (m, 1H), from 2.00 (s, 3H), 2,82 (s, 2H), is 4.21 (s, 2H), 7,27 (m, 5H), 7,76 (m, 4H), a 7.62 (m, 2H), 10,1 (s, 1H), 11,93 (users, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 1,619 microns.

Example 5-22: 4-((N-(4-terbisil)phenylsulfonyl)methyl)cyclohexanecarbonyl acid

Was obtained as in example 5-11 from benzosulfimide, 4-(aminomethyl)cyclohexanecarboxylic acid and 1-(methyl bromide)-4-fervently. MS (M+H, 406); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 0.77 (m, 2H), were 0.94 (m, 2H), and 0.98 (m, 2H), 1,50 (m, 2H), 1,5 (m, 2H), 1.77 in (m, 1H), 2,80 (d, 2H), 4,28 (s, 2H), 7,20 (m, 2H), 7,38 (m, 2H), 7.62mm (m, 2H), of 7.70 (m, 1H), 7,88 (m, 2H), 11,93 (users, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0,240 microns.

Example 5-23: 4-(N-(cyclohexylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and cyclohexylethylamine. MS (M-H, 416,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 0,63 (m, 2H), 0.87 (m, 3H), were 0.94 (m, 1H), 1,25-of 1.52 (m, 5H), to 1.70 (m, 2H), 2,86 (m, 2H), 3,70 (s, 3H), 4,22 (s, 2H), 6,84 (m, 2H), 7,16 (m, 2H), to $ 7.91 (m, 2H), 7.62mm (m, 2H), 8,09 (m, 2H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 3,47 microns.

Example 5-24: 4-((N-(4-terbisil)-1-phenylmethylsulfonyl)methyl)cyclohexanecarbonyl acid

Was obtained as in example 5-10 from phenylmethanesulfonyl and methyl-4-(aminomethyl)cyclohexanecarboxylate and 1-(methyl bromide)-4-fervently. MS (M-H, 418);1H NMR (400 MHz, DMSO-d6): δ (million parts: 0,639 (m, 2H), 0,897 (m, 2H), to 1.034 (m, 1H), 1,467 (d, OSiR., 2H, J=11.2 Hz), 1,709 (d, OSiR., 2H, J=11.2 Hz), 1,961 (m, 1H), 2,828 (d, 2H, J=7,6 Hz)4,207 (s, 2H), 4,449 (s, 2H), 7,155 (t, 2H, J=9,2 Hz), 7,377 (m, 7H), 12 (C, user., 1H)

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 amounted to 9.57 microns.

Example 5-25: 4-(N-(2-cyanobenzyl)-N-(4-methoxybenzo the l)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and alpha-bromo-o-tolunitrile. MS (M-H, 435,1);1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,664 (s, 2H), 4,348 (s, 2H), 4,498 (s, 2H), 6,728 (d, 2H, J=8,4 Hz)7,036 (d, 2H, J=8,4 Hz), 7,352 (t, 2H, J=9,2 Hz), 7,548 (t, 1H, J=7,6 Hz)7,640 (d, 1H, J=7,6 Hz)8,003 (d, 2H, J=8 Hz), 8,139 (d, 2H, J=8,4 Hz)13,559 (with, of user., 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was br4.61 microns.

Example 5-26: 4-(N-(4-acetamidobenzoyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and N-(4-(chloromethyl)phenyl)ndimethylacetamide. MS (M-H, 467,1);1H NMR (400 MHz, DMSO-d6): δ (million parts: 2,0 (s, 3H), of 3.69 (s, 3H), 4,23 (s, 4H), 6,78 (d, 2H, J=7,6 Hz), 6,98 (m, 4H), 7,41 (d, 2H, J=8 Hz), 7,94 (d, 2H, J=8 Hz), of 8.09 (d, 2H, J=8 Hz), for 9.90 (s, 1H).

Example 5-27: 4-(N-(furan-3-ylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

110 mg of 1-(furan-3-yl)-N-(4-methoxybenzyl)methanamine (example 5-27a) was mixed with methyl-4-(chlorosulfonyl)benzoate (example 5-10c) (117 mg, 0.5 mmol) and triethylamine (100 μl) in DHM (5 ml). The mixture was stirred overnight at ambient temperature and concentrated. The residue was re-dissolved in ethyl acetate (20 ml), washed with 1N HCl (aqueous, 2 ml), and then in the Oh (5 ml) and with brine (5 ml), then was dried over magnesium sulfate. The crude product was purified by the method of preparative TLC (40% ethyl acetate/hexane) to obtain methyl 4-(N-(furan-3-ylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoate as a white solid. Saponification carried out as in example 5-10, allowed to obtain 4-(N-(furan-3-ylmethyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid as a white crystalline solid (68 mg, 64% yield). MS (M-H, 400,1);1H NMR (400 MHz, DMSO-d6): δ (million parts: and 3.72 (s, 3H), of 4.13 (s, 2H), 4.26 deaths (s, 2H), of 5.99 (s, 1H), 6.87 in (m, 4H), 6.87 in (d, 2H, J=8,8 Hz), 7,13 (d, 2H, J=8,8 Hz), 7,39 (s, 1H), 7,49 (s, 1H), 7,95 (d, 2H, J=8,4 Hz), of 8.09 (d, 2H, J=8,8 Hz).

Example 5-27a: 1-(furan-3-yl)-N-(4-methoxybenzyl)methanamine

A mixture of 3-furaldehyde (5 mmol, 437 μl) and (4-methoxyphenyl)methanamine in MeOH (20 ml) was stirred at ambient temperature overnight, then was slowly added sodium borohydride (300 mg, 7,89 mmol). The resulting mixture was stirred at room temperature for 15 minutes and extinguished NaOH (1 N, water). The methanol was removed in vacuum and then the resulting suspension was re-dissolved in ethyl acetate, washed with water, with brine, dried over sodium sulfate and concentrated. After purification by chromatography on silica gel (ethyl acetate:hexane 7:3) was obtained 1-(furan-3-yl)-N-(4-methoxybenzyl)methanamine in the form of oil. MS (M+H 218,10);1H NMR (400 MHz, CDCl3): δ million parts of 3.64(s, 2H), 3,74 (s, 2H), 3,80 (s, 3H), to 6.39 (m, 1H), 6,86 (m, 1H), to 6.88 (m, 1H), 7.23 percent (m, 1H), 7,25 (m, 1H), 7,35 (m, 1H), 7,38 (m, 1H).

Example 5-28: 4-(N,N-dibenzylamino)benzoic acid

Was obtained as in example 5-27 from dibenzylamine and methyl-4-(chlorosulfonyl)benzoate (example 5-10c). MS (M-H, 380,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts: to 4.52 (s, 4H), 7,10 (m, 4H), 7,25 (m, 6H), of 8.00 (d, 2H, J=8,4 Hz), 8,15 (d, 2H, J=8,4 Hz), and 13.5 (with, of user., 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 7,74 microns.

Example 5-29: 4-(N-(4-methoxybenzyl)-N-methylsulfonyl)benzoic acid

Was obtained as in example 5-27 from 1-(4-methoxyphenyl)-N-methylmethanamine and methyl-4-(chlorosulfonyl)benzoate (example 5-10c). MS (M-H, 335,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts of 2.51 (s, 3H), 3,71 (s, 3H), of 4.05 (s, 2H), to 6.88 (d, 2H), 7,18 (d, 2H), to $ 7.91 (d, 2H); 8,13 (d, 2H). Elemental analysis: (experimental): C 57,47%, N of 4.77% and N or 4.31%; and (theoretically): C 57,30%, N 5,11% and N 4,18%.

Example 5-30: 4-(N,N-bis(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-27 from bis(4-methoxybenzyl)amine and methyl-4-(chlorosulfonyl)benzoate (example 5-10c). MS (M-H, 440,1); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,68 (s, 6H), 4,20 (s, 4H), 6,77 (d, 4H, J=10 Hz), 6,98 (d, 4H, J=10 Hz), 7,92 (DD, 2H, J=8 Hz), of 8.06 (DD, 2H, J=8 Hz). Elemental analysis: (experimental): C 62,45%, N 5,19% and N 3,06%; and (theoretically): C 62,57%, N of 5.25% and N 3,17%./p>

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 amounted to 4.14 μm.

Example 5-31: 4-(N-(2-terbisil)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and 1-(methyl bromide)-2-fervently. 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,66 (s, 3H), 4,29 (s, 2H), 4,37 (s, 2H), 6,72 (d, 2H, J=8 Hz), 7,01-7,03 (m, 6H), to 7.93 (d, 2H, J=8 Hz), 8,08 (d, 2H, J=8 Hz).

Example 5-32: 4-(N-(2,5-diferensial)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and 2-(methyl bromide)-1,4-diferente. 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,66 (s, 3H), or 4.31 (s, 2H), 4,33 (s, 2H), 6,74-7,06 (m, 7H), to 7.95 (d, 2H, J=8 Hz), of 8.09 (d, 2H, J=8 Hz).

Example 5-33: 4-(N-(2,3-diferensial)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b) and 1-(methyl bromide)-2,3-diferente. 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,30 (s, 3H), 4,29 (s, 2H), 4,32 (s, 2H), 6.87 in (d, 2H, J=8 Hz), 7,02-7,20 (m, 5H), to 7.95 (d, 2H, J=8 Hz). with 8.05 (d, 2H, J=8 Hz).

Example 5-34: 4-(N-(3-methoxybenzyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxy shall ensil)sulfamoyl)benzoate (example 5-10b) and 3-methoxybenzylamine. MS (M-H, 440,50); 1H NMR (400 MHz, DMSO-d6): δ (million parts: to 3.58 (s, 3H), 3,68 (s, 3H), 4,24 (s, 2H), 4,25 (s, 2H), 6,50 (s, 1H), only 6.64 (d, J=4 Hz, 1H), 6.73 x (m, 1H), 6,77 (d, J=8 Hz, 2H), 7,00 (d, J=8 Hz, 2H), 7,12 (t, J=8 Hz, 1H), 7,94 (d, J=8 Hz, 2H), of 8.09 (d, J=8 Hz, 2H), 13,49 (s, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 amounted to 2.46 microns.

Example 5-35: 4-(N-benzyl-N-(4-methoxyphenyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(N-(4-methoxyphenyl)sulfamoyl)benzoate (example 5-35a) and benzylbromide. MS (M-H, 396); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,66 (s, 3H), and 4.75 (s, 2H), 6,76 (d, J=8 Hz, 2H), make 6.90 (d, J=8 Hz, 2H), 7.23 percent (m, 5H), 7,74 (d, J=8 Hz, 2H), 8,11 (d, J=8 Hz, 2H), 13,51 (s, 1H).

Example 5-35a: Methyl-4-(N-(4-methoxyphenyl)sulfamoyl)benzoate

To 4-methoxybenzylamine (580 mg, 4,71 mmol) and triethylamine (1,48 ml and 10.7 mmol) in dichloromethane (10 ml) was added methyl-4-(chlorosulfonyl)benzoate (1,00 g, to 4.28 mmol). This mixture was stirred for 16 hours at room temperature. The reaction mixture was diluted with dichloromethane (50 ml) and washed successively with water, 10% citric acid and brine. The organic phase was dried over sodium sulfate and concentrated on a rotary evaporator. The resulting crude substance was chromatographically on silica gel using 100% dichloromethane as eluent, obtaining methyl-4-(N-(4-methoxyphenyl)sulfamoyl)benzoe is in the form of a white crystalline solid (400 mg, 30% yield).

Example 5-36: 4-(N-(3,4-diferensial)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from N-(3,4-diferensial)(4-methoxyphenyl)methanamine (example 5-36a) and methyl-4-(chlorosulfonyl)benzoate (example 5-10c). MS (M-H, 446); 1H NMR (400 MHz, DMSO-d6): δ million parts of 3.69 (s, 3H), 4,20 (s, 4H), 6.73 x (d, J=8,8 Hz, 2H), 6,93 (m, 2H), 6,98 (d, J=8,8 Hz, 2H), 7,22 (m, 1H), 7,74 (d, J=8,4 Hz, 2H), to 7.99 (d, J=8,4 Hz, 2H).

Example 5-36a: N-(3,4-diferensial)(4-methoxyphenyl)methanamine

To (4-methoxyphenyl)methanamine (1.77 ml of 13.6 mmol) and acetic acid (2.7 ml, 45 mmol) in dichloromethane (15 ml) was added 3,4-differentally (1.0 ml, the remaining 9.08 mmol). This mixture was heated in a microwave oven at 100ºC for 15 minutes the Reaction mixture was cooled to room temperature and the portions was added macroporous laborgerate resin (9,8 g, 22.7 mmol). This mixture was stirred at room temperature for 16 hours. The resin was filtered and washed with dichloromethane, and the organic matter was washed with saturated sodium bicarbonate until the cessation of gassing. Organic matter was dried over sodium sulfate and concentrated on a rotary evaporator. The resulting crude substance was purified by chromatography on silica gel using a gradient of methanol-dichloromethane as eluent and was obtained N-(3,4-diferensial)(4-methoxyphenylethylamine in the form of a yellowish oil (1.9 g, 80% yield). MS (M+H, 264); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 2,63 (Sirs, 1H), of 3.56 (s, 2H), 3,61 (s, 2H), 3,71 (s, 3H), at 6.84 (d, J=8,8 Hz, 2H), 7,14 (m, 1H), 7,21 (d, J=8,4 Hz, 2H), 7,34 (m, 2H).

Example 5-37: 4-((N-(4-terbisil)-4-methylphenylsulfonyl)methyl)benzoic acid

Was obtained as in example 5-11 4-(aminomethyl)phenylcarbinol acid, 4-methylbenzol-1-sulphonylchloride and 4-ftorangidridy. 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,11 (s, 3H), is 4.21 (s, 2H), 4,24 (s, 2H), 6,94-was 7.08 (m, 6H), 7,40-7,42 (d, 2H, J=8 Hz), 7,63 (d, 2H, J=8 Hz), 7,73 (d, 2H, J=8 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0,054 microns.

Example 5-38: 4-((4-carboxy-N-(4-methoxybenzyl)phenylsulfonyl)methyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(methyl bromide)benzoate and methyl 4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b). 1H NMR (400 MHz, DMSO-d6): δ (million parts: the 3.65 (s, 3H), 4,24 (s, 2H), 4,33 (s, 2H), of 6.71 (d, 2H, J=8gts), to 6.95 (d, 2H, J=8 Hz), 7,14 (d, 2H, J=8 Hz), 7,73 (d, 2H, J=8 Hz), 7,89 (d, 2H, J=8 Hz), of 8.06 (d, 2H, J=8 Hz).

Example 5-39: 4-(N-benzyl-N-(3,4-dimethoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from (3,4-acid)methanamine, methyl-4-(chlorosulfonyl)benzoate (example 5-10c) and benzylchloride. MS (M-H, 440,10); 1H NMR (400 MHz, CD3OD): δ million parts: 3,59 (who, 3H), 3,76 (s, 3H), 4,30 (s, 2H), 4,36 (s, 2H), 6,51 (d, 1H, J=1.7 Hz), 6,60 (m, 1H), 6,76 (d, 1H, J=8,2 Hz), 7,12 (m, 2H), 7,21 (m, 3H), 7,95 (d, 2H, J=8.6 Hz), 8,18 (d, 2H, J=8.6 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0,678 microns.

Example 5-40: 4-(N-(3,4-dimethoxybenzyl)-N-(4-methoxybenzyl) sulfamoyl)benzoic acid

Was obtained as in example 5-10 from (3,4-acid)methanamine, methyl-4-(chlorosulfonyl)benzoate (example 5-10c) and 4-methoxybenzylamine. MS (M-H, 470,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,49 (s, 3H), to 3.67 (s, 3H), 3,68 (s, 3H), 4,20 (s, 2H), 4,22 (s, 2H), 6,41 (d, 1H, J=1.4 Hz), to 6.58 (DD, 1H, J1=8.2 Hz, J2=1.4 Hz), 6,78 (m, 3H), 7,02 (d, 2H, J=8.6 Hz), 7,94 (d, 2H, J=8,4 Hz), of 8.09 (d, 2H, J=8,4 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 1.47 ám.

Example 5-41:4-(N-(3-fluoro-4-methoxybenzyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from 4-(methyl bromide)-2-fluoro-1-methoxybenzene and methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b). MS (M-H, 458,10); 1H NMR (400 MHz, CD3OD): δ million parts of 3.73 (s, 3H), of 3.80 (s, 3H), 4,24 (s, 2H), 4,28 (s, 2H), 6.75 in (m, 4H), to 6.88 (m, 1H), 6,97 (d, 2H, J=8.6 Hz), 7,88 (d, 2H, J=8,3 Hz)to 8.14 (d, 2H, J=8,3 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 1.11 mm.

Example 5-42: 4-((4-hydroxy-N-(2-methoxybenzyl)FeNi is sulphonamido)methyl)benzoic acid

4-(N-(2-methoxybenzyl)sulfamoyl)phenyl acetate (example 5-42a) (50 mg, 0.15 mmol) was dissolved in acetone (1.0 ml)and then was added cesium carbonate (97 mg, 0.30 mmol) and methyl-4-(methyl bromide)benzoate (38 mg, 0,17 mmol). The mixture was stirred at room temperature overnight and then the inorganic salts were filtered off. The acetone was removed under vacuum, and the residue was re-dissolved in ethyl acetate and washed with water and then brine. The organic phase was dried over magnesium sulfate and concentrated. The crude product was purified by the method of column chromatography using ethyl acetate/hexane as eluent to obtain methyl 4-((4-acetoxy-N-(2-methoxybenzyl)phenylsulfonyl)methyl)benzoate.

Methyl-4-((4-acetoxy-N-(2-methoxybenzyl)phenylsulfonyl)methyl)benzoate (crude) was dissolved in THF (1.0 ml) and was treated with aqueous NaOH (1N, 2.0 ml, 2.0 mmol). The mixture was boiled under reflux for one hour. Upon completion of the THF was removed under vacuum, and the resulting aqueous solution was acidified using 6 N aqueous HCl to pH ~3. The aqueous phase was extracted with EtOAc (2×15 ml) and the combined organic phases are washed with water, brine, dried over Na2SO4and concentrated. The crude product was purified by the method of reversed-phase HPLC with getting to 10.8 mg of the above compound (15% yield in two articles which dealt with). MS (M-H, 426,1); 1H NMR (400 MHz, acetone-d6): δ (million parts: the 3.65 (s, 3H), 4,37 (s, 2H), 4,43 (s, 2H), 6,79 (m, 2H), 7,01 (d, 2H, J=8.0 Hz), 7,17 (m, 2H), 7,28 (d, 2H, J=7.9 Hz), 7,73 (d, 2H, J=8.0 Hz), 7,87 (d, 2H, J=7.9 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 were $ 2.56 ám.

Example 5-42a: 4-(N-(2-methoxybenzyl)sulfamoyl)phenylacetate

A solution of 4-(chlorosulfonyl)phenylacetate (example 5-42b) (531 mg, 2,265 mmol) in 5.0 ml of dichloromethane, was cooled to 0ºC in an ice bath. Was added (2-methoxyphenyl)methanamine (325 μl, 2,492 mmol) and triethylamine (347 μl, 2,492 mmol). Then bath with ice was removed, and the mixture was heated to ambient temperature and was stirred for 2 hours. The reaction mixture was concentrated and the crude product is purified by the method of column chromatography (hexane/ethyl acetate=90/10-30/70) to obtain pure 4-(N-(2-methoxybenzyl)sulfamoyl)phenylacetate (743 mg, 89%) as a white solid. MS (M+H, 336,1) 1H NMR (400 MHz, CDCl3): δ million parts: 2,32 (s, 3H), 3,71 (s, 3H), 4,18 (d, 2H, J=5.8 Hz), further 5.15 (t, 1H, J=5.8 Hz), of 6.71 (d, 1H, J=8,2 Hz), 6,82 (ushort, 1H, J=7.4 Hz), 7,07 (userd, 1H, J=7.4 Hz), 7,10 (d, 2H, J=8.7 Hz), 7,19 (ushort, 1H, J=7,8 Hz), 7,74 (d, 2H, J=8.7 Hz).

Example 5-42b: 4-(chlorosulfonyl)phenylacetate

6,285 g (36,08 mmol) 4-hydroxybenzenesulfonate was dissolved in a mixture of 30 ml of acetic anhydride and 15 ml of acetic acid and boiled under reflux for 6 hours. Volatile components in perivale and placed in a deep vacuum on during the night. The resulting crude product was dissolved in 100 ml DHM and processed 4,72 ml oxalicacid (54,12 mmol) and 139 μl DMF) (1,804 mmol) at 0ºC. Continued stirring until gas evolution stops, then the reaction mixture was concentrated and re-dissolved in EtOAc. The organic phase was washed twice with 2 N H2SO4and dried with brine and MgSO4. After concentration was received 7,067 g of 4-(chlorosulfonyl)phenylacetate as a dark thick oil, which eventually hardened (83% yield in two stages).

1H-NMR (400 MHz, CDCl3): δ million parts of 2.35 (s, 3H), 7,37 (d, 2H, J=8,9 Hz), of 8.06 (d, 2 H, J=8,9 Hz).13C-NMR (100 MHz, CDCl3): δ, mln pieces: 21,13, 123,05, 128,91, 141,15, 155,80, 168,29.

Example 5-43: 4-((4-hydroxy-N-(3-methoxybenzyl)phenylsulfonyl)methyl)benzoic acid

Was obtained as in example 5-42 from (3-methoxyphenyl)methanamine and methyl-4-(methyl bromide)benzoate. MS (M-H, 426,10); 1H NMR (400 MHz, acetone-d6): δ (million parts: 3,66 (s, 3H), 4,32 (s, 2H), and 4.40 (s, 2H), 6,65 (users, 1H), 6.73 x (m, 2H), 7,05 (d, 2H, J=8.5 Hz), 7,12 (t, 1H, J=8.0 Hz), 7,27 (d, 2H, J=8.0 Hz), 7,82 (d, 2H, J=8.5 Hz), 7,88 (d, 2H, J=8.0 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0,188 microns.

Example 5-44: 4-((4-hydroxy-N-(4-methoxybenzyl)phenylsulfonyl)methyl)benzoic acid

Got the AK in example 5-42 (4-methoxyphenyl)methanamine and methyl-4-(methyl bromide)benzoate. MS (M-H, 426,10); 1H NMR (400 MHz, acetone-d6): δ (million parts of 3.73 (s, 3H), 4,27 (s, 2H), 4,35 (s, 2H), 6,76 (d, 2H, J=8.0 Hz),? 7.04 baby mortality (d, 4H), from 7.24 (d, 2H, J=7.8 Hz), 7,79 (d, 2H, J=8,2 Hz), 7,88 (d, 2H, J=7,8 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 3,43 mm.

Example 5-45: 4-((N-(3-terbisil)-4-hydroxyphenylacetamide)methyl)benzoic acid

Was obtained as in example 5-42 from (3-forfinal)methanamine and methyl-4-(methyl bromide)benzoate. MS (M-H, 414,1), 1H NMR (400 MHz, acetone-d6): δ (million parts: 4,37 (s, 2H), 4,42 (s, 2H), 6,94 (m, 3H), 7,06 (d, 2H, J=8,3 Hz), 7,21 (m, 1H), 7,28 (d, 2H, J=7,6 Hz), 7,81 (d, 2H, J=8,3 Hz), 7,87 (d, 2H, J=7,6 Hz).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0,459 microns.

Example 5-46: N-benzyl-N-(4-methoxybenzyl)-4-(1H-tetrazol-5-yl)benzene-sulfonamide

N-benzyl-4-cyano-N-(4-methoxybenzyl)benzosulfimide (example 5-46a, 400 mg, 1 mmol) and azide trimethylurea (400 mg, 2 mmol) was dissolved in toluene (10 ml) and subjected to microwave radiation at about 150ºc for 3 hours. Added additional 2 equivalent azide trimacinolone and the reaction mixture was subjected to microwave radiation at about 150ºc for 3 more hours. The mixture was cooled and filtered to obtain the crude tetrazole tin, which was hydrolyzed in MEOH/kontsentrirovano what I HCl (50 ml: 20 ml). Added water and the resulting precipitate was collected by filtration. The product was recrystallized from absolute ethanol and water to obtain N-benzyl-N-(4-methoxybenzyl)-4-(1H-tetrazol-5-yl)benzene-sulfonamida in the form of a white solid. MS (M+H 436,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts: the 3.65 (s, 3H), 4,27 (s, 2H), 4,30 (s, 2H), 6.75 in (d, J=8,4 Hz, 2H), 6,98 (d, J=8,4 Hz, 2H), was 7.08 (m, 2H), 7,20 (m, 3H), with 8.05 (d, J=8,4 Hz, 2H), by 8.22 (d, J=9,2 Hz, 2H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 made 3.67 ám.

Example 5-46a: N-benzyl-4-cyano-N-(4-methoxybenzyl)benzosulfimide

4-Cyanobenzoyl-1-sulphonylchloride (600 mg, 3 mmol) was added to a solution of N-benzyl-1-(4-methoxyphenyl)methanamine (example 5-46b, 750 mg, 3.3 mmol) and triethylamine (500 mg, 3.6 mmol) in dichloromethane (15 ml). The reaction mixture was stirred at ambient temperature for 4 hours, then concentrated on a rotary evaporator. The crude substance was purified on silica gel to obtain N-benzyl-4-cyano-N-(4-methoxybenzyl)benzene-sulfonamidain the form of a white solid. 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,68 (s, 3H), 4.26 deaths (s, 2H), or 4.31 (s, 2H), 6.75 in (d, J=8,4 Hz, 2H), 6,97 (d, J=9,2 Hz, 2H), 7,07 (m, 2H), 7,21 (m, 3H), 7,98 (d, J=8,4 Hz, 2H), 8,04 (d, J=8,8 Hz, 2H).

Example 5-46b: N-benzyl-1-(4-methoxyphenyl)methanamine

4-Methoxybenzaldehyde (5 g, 35 mmol) and benzylamine (3.8 g, 35 mmol) was added to triacet is cyberguide sodium (10.4 g, 49 mmol) in dichloroethane (125 ml). The reaction mixture was stirred at ambient temperature for 2 hours, then concentrated. The mixture was diluted with dichloromethane (200 ml), washed with saturated aqueous sodium bicarbonate (200 ml), brine (200 ml) and dried over magnesium sulfate. The crude amine was concentrated and purified by chromatography on silica gel (70% ethyl acetate in hexane) to give N-benzyl-1-(4-methoxyphenyl)methanamine in the form of oil. 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,59 (s, 2H), to 3.64 (s, 2H), 3,71 (s, 3H), 6,86 (d, J=8,8 Hz, 2H), 7,28 (m, 7H).

Example 5-47: 2-(4-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)phenyl)acetic acid

Was obtained as in example 5-27 of methyl-2-(4-(chlorosulfonyl)phenyl)acetate (example 5-47a), 4-methoxybenzaldehyde and benzylamine. MS (M-H, 424,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,66 (s, 3H), and 3.72 (s, 2H), 4,18 (s, 2H), 4,22 (s, 2H), 6.73 x (d, J=8,4 Hz, 2H), 6,91 (d, J=8,4 Hz, 2H), 7,02 (m, 2H), 7,19 (m, 3H), 7,49 (d, J=8,4 Hz, 2H), 7,80 (d, J=8 Hz, 2H).

Example 5-47a: Methyl-2-(4-(chlorosulfonyl)phenyl)acetate

2-(4-(Chlorosulfonyl)phenyl)acetic acid (600 mg, 2.6 mmol) was added to thionyl chloride (3 ml) and heated at 80°C for 1 hour. The reaction mixture was concentrated, cooled to 0ºC in an ice bath and added dropwise ice-cold methanol. The mixture was stirred for 30 minutes and then concentrated to obtain methyl 2-(4-(chlorosulfonyl)f the Nile)acetate in the form of oil. 1H NMR (400 MHz, DMSO-d6): δ (million parts of 3.57 (s, 3H), of 3.65 (s, 2H), 7,21 (d, J=8 Hz, 2H), 7,55 (d, J=8 Hz, 2H).

Example 5-48: 4-((N-benzyl-4-carboxyphenylsulfate)methyl)benzoic acid

Was obtained as in example 5-10 from methyl-4-(chlorosulfonyl)benzoate (example 5-10c), benzylamine and methyl-4-(methyl bromide)benzoate. MS (M+H, 426,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts: the 4.29 (s, 2H), 4,34 (s, 2H), 7,03 (m, 2H), 7,12 (m, 2H), 7,68 (d, J=8 Hz, 2H), 7,94 (d, J=8,8 Hz, 2H), of 8.06 (d, J=8,8 Hz, 2H), 13,03 (s, 2H).

Example 5-49: 4-(benzyl(4-methoxybenzyl)carbarnoyl)benzoic acid

Was obtained as in example 5-27 4-(chlorocarbonyl)benzoate and N-benzyl-1-(4-methoxyphenyl)methanamine. 1H NMR (400 MHz, DMSO-d6): δ (million parts: and 3.72 (d, J=7,6 Hz, 3H), 4.26 deaths (s, 1H), 4,30 (s, 1H), 4,50 (s, 1H), 4,55 (s, 1H), 6.89 in (m, 2H), 7,02 (m, 1H), 7,10 (m, 1H), 7,10 (m, 1H), 7,20 (m, 1H), 7,28 (m, 4H), 7,55 (m, 2H), of 7.96 (m, 2H), 13,13 (s, 1H).

Example 5-50: 4-(N-(4-fluoro-3-methoxybenzyl)-N-(4-methoxybenzyl)sulfamoyl)benzoic acid

Was obtained as in example 5-10 from 4-(methyl bromide)-4-fluoro-3-methoxybenzene and methyl-4-(N-(4-methoxybenzyl)sulfamoyl)benzoate (example 5-10b). MS (M-H, 458,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,59 (s, 3H), to 3.67 (s, 3H), 4,25 (s, 2H), 4,25 (s, 2H), 6,62 (m, 2H), 6,77 (d, J=8,4 Hz, 2H), 7,02 (m, 3H), of 7.97 (d, J=8 Hz, 2H), 8,01 (d, J=8,8 Hz, 2H), 13,49 (s, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 1.65 μm.

p> Additional connections are experimentally investigated and found that they have a relatively high level of effectiveness as inhibitors of the bitter taste receptor hT2R14. The results of this study are presented below in table C.

Table C
No. of connectionsConnectionhT2R14 IC50(µm)
5-51
4-((N-(4-terbisil)thiophene-2-sulfonamide)methyl)
cyclohexanecarbonyl acid
0,342
5-52
4-((N-(4-terbisil)pyridine-3-sulfonamide)methyl)
cyclohexanecarbonyl acid
8,434
5-53
4-((N-(4-terbisil)pyridine-2-sulfonamide)methyl)
cyclohexanecarbonyl acid
5-54
4-(N-(furan-2-ylmethyl)sulfamoyl)benzoic acid

5-55
4-(N-benzyl-N-methylsulfonyl)benzoic acid
5-56
4-(N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
5-57
3-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)
propanoic acid
5-58
4-(N-(2-methoxybenzyl)-N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
2,943
5-59
3-(4-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)
phenyl)propanoic acid

5-60
2-(N-benzyl-N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
5-61
4-(N-(4-methoxybenzyl)-N-(thiophene-2-ylmethyl)sulfamoyl)benzoic acid
5-62
4-(N-(4-methoxybenzyl)-N-(thiophene-3-ylmethyl)sulfamoyl)benzoic acid
5-63
4-(N-(4-methoxybenzyl)-N-((5-methylfuran-2-yl)methyl)sulfamoyl)benzoic acid

5-64
4-(N-(4-methoxybenzyl)-N-(2-methylbenzyl)sulfamoyl)
benzoic acid
5-65
4-(N-(4-methoxybenzyl)-N-(3-methylbenzyl)sulfamoyl)
benzoic acid
5-66
4-(N-(4-methoxybenzyl)-N-(4-methylbenzyl)sulfamoyl)
benzoic acid
5-67
4-(N-(3,5-dimethoxybenzyl)-N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
12,141
5-68
4-(N-(2-Chlorobenzyl)-N-(4-methoxybenzyl)sulfamoyl)
Ben is oyna acid
7,044

5-69
4-(N-(4-terbisil)-N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
8,283
5-70
4-(N-benzyl-N-(4-methylbenzyl)sulfamoyl)
benzoic acid
5,394
5-71
4-(N-benzyl-N-(2-methoxybenzyl)sulfamoyl)
benzoic acid
5,061
5-72
4-(N-benzyl-N-(2-terbisil)sulfamoyl)
benzoic acid
6,164
5-73
4-(N-benzyl-N-(3-methylbenzyl)sulfamoyl)
benzoic acid
2,290

5-74
4-(N-benzyl-N-(3-methoxybenzyl)sulfamoyl)
benzoic acid
5,991
5-75
4-(N-benzyl-N-(3-cyanobenzyl)sulfamoyl)
benzoic acid
5-76
3-((N-benzyl-4-carboxyphenylsulfate)
methyl)benzoic acid
5-77
4-(N-benzyl-N-(4-chlorbenzyl)sulfamoyl)
benzoic acid
3,977
5-78
4-(N-benzyl-N-(4-(methylthio)benzyl)sulfamoyl)
benzoic acid
2,458

5-79
4-(N-benzyl-N-(3-terbisil)sulfamoyl)
benzoic acid
2,310
5-80
4-(N-benzyl-N-(2-methylbenzyl)sulfamoyl)
benzoic acid
2,926
5-81
4-(N-benzyl-N-(4-methoxyphenethyl)sulfamoyl)
benzoic acid
5-82
3-((4-carboxy-N-(4-methoxybenzyl)
phenylsulfonyl)methyl)
benzoic acid

5-83
4-(N-benzyl-N-(2-Chlorobenzyl)sulfamoyl)
benzoic acid
4,486
From 5 to 84
4-(N-benzyl-N-(3-Chlorobenzyl)sulfamoyl)
benzoic acid
4,286
5-85
4-(N-benzyl-N-(4-active compounds)sulfamoyl)
benzoic acid
8,580
5-86
4-(N-(4-methoxybenzyl)-N-(pyridine-4-ylmethyl)sulfamoyl)benzoic acid
5-87
4-(N-(4-methoxybenzyl)-N-(pyridine-2-ylmethyl)sulfamoyl)benzoic acid

5-88
4-(N-(4-methoxybenzyl)-N-(pyridine-3-ylmethyl)sulfamoyl)benzoic acid
5-89
4-(N-benzyl-N-(4-methoxy-3-methylbenzyl)sulfamoyl)
benzoic acid
1,995
5-90
4-(N-benzyl-N-((6-methoxypyridine-3-yl)methyl)sulfamoyl)benzoic acid
3,258
5-91
4-(N-benzyl-N-(4-cyanobenzyl)sulfamoyl)
benzoic acid
of 1, 848

5-92
4-(N-benzyl-N-(2-cyanobenzyl)sulfamoyl)
benzoic acid
4,627
5-93
4-(N-(benzo[d][1,3]dioxol-5-ylmethyl)-N-benzylcarbamoyl)benzoic acid
2,167
5-94
4-(3-benzyl-3-(4-methoxybenzyl)ureido)
benzoic acid
5-95
4-(N-(4-methoxybenzyl)-N-(1-phenylethyl)sulfamoyl)
Ben is oyna acid
1,397

5-96
4-(N-(4-methoxybenzyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)benzoic acid
2,415
5-97
4-(N-((4-methoxyphenyl)(phenyl)methyl)
sulfamoyl)benzoic acid
5-98
4-(4-benzyl-3-(4-methoxybenzyl)-2,5-dioxoimidazolidin-1-yl)benzoic acid
5-99
4-(N-benzyl-N-(4-hydroxybenzyl)sulfamoyl)
benzoic acid
5,207

5-100
4-((N-(2-methoxybenzyl)
phenylsulfonyl)methyl)
benzoic acid
1,294
5-101
4-((N-(3-methoxybenzyl)
phenylsulfonyl)methyl)
benzoic acid
0,345
5-102
4-((N-(4-methoxybenzyl)
phenylsulfonyl)methyl)
benzoic acid
2,219
5-103
4-((N-(2-terbisil)phenylsulfonyl)methyl)benzoic acid
0,429

5-104
4-((N-(3-terbisil)phenylsulfonyl)methyl)benzoic acid
0,406
5-105
4-((N-(4-terbisil)phenylsulfonyl)methyl)benzoic acid
0,935
5-106
4-(2-methoxy-5H-dibenzo[c,e]azepin-6(7H)-ylsulphonyl)benzoic acid
5-107
4-((N-(4-terbisil)-4-hydroxyphenylacetamide)
methyl)benzoic acid
4,086

5-108
4-(N-benzyl-N-(4-ethoxy ensil)sulfamoyl)
benzoic acid
5-109
4-(N-benzyl-N-(4-propoxyphenyl)sulfamoyl)
benzoic acid
5-110
4-(N-benzyl-N-(4-isopropoxyphenyl)sulfamoyl)benzoic acid
5-111
4-((3-(3-chloro-4-were)-1-(4-methoxybenzyl)touraid)
methyl)cyclohexanecarbonyl acid
6,136

tr>
5-112
4-((1-(4-methoxybenzyl)-3-(2-methoxyphenyl)touraid)methyl)cyclohexanecarbonyl acid
8,852
5-113
4-((1-(4-active compounds)-3-o-tolylthiourea)methyl)
cyclohexanecarbonyl acid
5,018
5-114
4-((3-(4-chloro-3-were)-1-(2-methylbenzyl)touraid)methyl)
cyclohexanecarbonyl acid
4,872
5-115
ethyl 4-((4-acetamido-N-benzilpenitsillino)
methyl)cyclohexanecarboxylate
0,334

5-116
ethyl 4-((N-(3-bromobenzyl)-2,3-dihydrobenzo[b][1,4]dioxin-6-sulfonamide)methyl)
cyclohexanecarboxylate
2,567
5-117
ethyl 4-((N-(2-Chlorobenzyl)-4-methylphenylsulfonyl)methyl)cyclohexanecarboxylate
2,816
5-118
ethyl 4-((N-benzyl-4-bromophenylacetate)methyl)
cyclohexanecarboxylate
2,344
5-119
ethyl 4-((N-benzyl-4-chlorophenylsulfonyl)methyl)
cyclohexanecarboxylate
0,672

5-121
5-120mm
4-((N-benzyl-4-chlorophenylsulfonyl)methyl)
benzoic acid
0,394

4-(N-(2-methoxybenzyl)-N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
2,943
5-122
N-benzyl-3-methoxy-N-(4-methoxybenzyl)
benzosulfimide
2,219
5-123
4-((N-(4-terbisil)-4-methoxybenzenesulfonamide)
methyl)cyclohexanecarbonyl acid
0,092

5-124
4-((4-acetamido-N-(4-terbisil)phenylsulfonyl)methyl)cyclohexanecarbonyl acid
0,842
5-125
4-((4-amino-N-(4-terbisil)phenylsulfonyl)methyl)cyclohexanecarbonyl acid
1,651
5-126
4-((4-methyl-N-(4-methylbenzyl)
phenylsulfonyl)methyl)
cyclohexanecarbonyl acid
0,109

5-27
4-((N-(4-cyanobenzyl)-4-methylphenylsulfonyl)methyl)cyclohexanecarbonyl acid
0,499
5-128
4-((N-(3-methoxybenzyl)-4-methylphenylsulfonyl)methyl)cyclohexanecarbonyl acid
being 0.036
5-129
4-(N-(4-methoxybenzyl)-N-(2-methylbenzyl)sulfamoyl)
benzoic acid
5,316
5-130
4-(N-(2-Chlorobenzyl)-N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
7,044

5-131
4-(N-benzyl-N-(4-terbisil)sulfamoyl)
benzoic acid
8,283
5-132
4-(N-benzyl-N-(4-methylbenzyl)sulfamoyl)
benzoic acid
5,394
5-133
4-(N-benzyl-N-(2-methoxybenzyl)sulfamoyl)
benzo is Naya acid
5,061
5-134
4-(N-benzyl-N-(2-terbisil)sulfamoyl)
benzoic acid
6,164
5-135
4-(N-benzyl-N-(3-methylbenzyl)sulfamoyl)
benzoic acid
2,290

5-136
4-(N-benzyl-N-(3-methoxybenzyl)sulfamoyl)
benzoic acid
5,991
5-137
4-(N-benzyl-N-(4-chlorbenzyl)sulfamoyl)
benzoic acid
3,977
5-138
4-(N-benzyl-N-(4-(methylthio)benzyl)sulfamoyl)
benzoic acid
2,458
5-139
4-(N-benzyl-N-(3-terbisil)sulfamoyl)
benzoic acid
2,310
5-140
4-(N-benzyl-N-(2-methylbenzyl)sulfamoyl)
Ben is oyna acid
2,926

5-141
4-(N-benzyl-N-(2-Chlorobenzyl)sulfamoyl)
benzoic acid
4,486
5-142
4-(N-benzyl-N-(3-Chlorobenzyl)sulfamoyl)
benzoic acid
4,286
5-143
4-(N-benzyl-N-(4-active compounds)sulfamoyl)
benzoic acid
8,580
5-144
4-(N-benzyl-N-(4-methoxy-3-methylbenzyl)sulfamoyl)
benzoic acid
1,995

5-145
4-(N-benzyl-N-((6-methoxypyridine-3-yl)methyl)sulfamoyl)benzoic acid
3,258
5-146
4-(N-benzyl-N-(4-cyanobenzyl)sulfamoyl)
benzoic acid
of 1, 848
5-147
4-(N-benzyl-N-(2-cyanobenzyl)sulfamoyl)
benzoic acid
4,627
5-148
4-(N-(benzo[d][1,3]dioxol-5-ylmethyl)-N-benzylcarbamoyl)benzoic acid
2,167

5-149
4-(N-(4-methoxybenzyl)-N-(1-phenylethyl)sulfamoyl)
benzoic acid
1,397
5-150
4-(N-(4-methoxybenzyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)sulfamoyl)benzoic acid
2,415
5-151
4-(N-benzyl-N-(4-hydroxybenzyl)sulfamoyl)
benzoic acid
5,207
5-152
4-((N-(2-methoxybenzyl)phenylsulfonyl)methyl)benzoic acid
1,294

5-153
4-((N-(3-methoxybenzyl)
phenylsulfonyl)methyl)
benzoic acid is the
0,345
5-154
4-((N-(4-methoxybenzyl)
phenylsulfonyl)methyl)
benzoic acid
2,219
5-155
4-((N-(2-terbisil)phenylsulfonyl)methyl)benzoic acid
0,429
5-156
4-((N-(3-terbisil)phenylsulfonyl)methyl)benzoic acid
0,406

5-157
4-((N-(4-terbisil)phenylsulfonyl)methyl)benzoic acid
0,935
5-158
4-(2-methoxy-5H-dibenzo[c,e]azepin-6(7H)-ylsulphonyl)benzoic acid
3,571
5-159
4-((N-(4-terbisil)-4-hydroxyphenylacetamide)
methyl)benzoic acid
4,086
5-160
4-(N-(2,4-dif is orbenin)-N-(4-methoxybenzyl)sulfamoyl)
benzoic acid
1,255

5-161
4-(3-(4-methoxyphenyl)-3,4-dihydroisoquinoline-2(1H)-ylsulphonyl)benzoic acid
6,267
5-162
4-(6-methoxy-1-phenyl-3,4-dihydroisoquinoline-2(1H)-ylsulphonyl)benzoic acid
9,616
5-163
4-(7-methoxy-1-phenyl-3,4-dihydroisoquinoline-2(1H)-ylsulphonyl)benzoic acid
2,879

Example 6-1: (Z)-3-(5-(2,5-dimethoxyaniline)-4-oxo-2-thioxothiazolidin-3-yl)propanoic acid

3-(4-Oxo-2-thioxothiazolidin-3-yl)propanoic acid (200 mg, 1 mmol), 2,5-dimethoxybenzaldehyde (168 mg, 1 mmol) and piperidine (0.3 ml) were mixed in ethanol (3 ml) and subjected to microwave radiation at 100ºC for 10 minutes. The reaction mixture was cooled, the solid is collected by filtration, washed with ethyl acetate/hexane (1/1) and recrystallized from ethanol to obtain 85% of the output of the above-mentioned compound (309 mg, brown-orange solid) MS (M+H, 284); 1H NMR (400 MHz, DMSO-d6): δ (million parts: to 2.57 (t, 2H), 3,74 (s, J=6,8 Hz, 3H), of 3.84 (s, 3H), 4,19 (t, 2H), 6.90 to (s, 1H), 7,10 (s, 2H), 7,86 (s, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 amounted to 2.75 μm.

Example 6-2: (E)-2-cyano-3-(furan-2-yl)-N-(5-phenyl-1,3,4-thiadiazole-2-yl)acrylamide

5-Phenyl-1,3,4-thiadiazole-2-amine (600 mg, 3.3 mmol), zanoxolo acid (300 mg, 3.6 mmol) and EDC-HCl (861 mg, 4.5 mmol) was stirred in acetonitrile (15 ml) at room temperature for 2 hours. The mixture was diluted with aqueous 1N HCl and the aqueous phase is extracted with ethyl acetate. The combined organic extracts were dried over sodium sulfate, filtered and concentrated under vacuum. The residue was ground into powder with ethyl acetate/hexane (1/9) to obtain 2-cyano-N-(5-phenyl-1,3,4-thiadiazole-2-yl)ndimethylacetamide in the form of a white solid, which was used without further purification.

312 mg of 2-cyano-N-(5-phenyl-1,3,4-thiadiazole-2-yl)ndimethylacetamide and 150 μl furan-2-carbaldehyde were mixed in 2.5 ml of DMF and heated in a microwave oven at about 150ºc for 15 minutes was Added 5 ml of H2O and the precipitate was collected and washed with water (4×) to give the crude product. Recrystallization from ethanol was allowed to obtain 180 mg of the pure product as a brown solid. MS (M+H, 323); 1H NMR (400 MHz, DMSO-d6): δ (million parts: PC 6.82(m, 1H), the 7.43 (d, 1H), 7,49 (m, 4H), 7,87 (m, 2H), 8,15 (s, 1H), they were 8.22 (users, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 0,499 microns.

Example 6-3: N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1,4-dimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulfonamide

1,4-Dimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulphonylchloride (example 6-3a, 450 mg, 1.6 mmol) was added under stirring to a solution of 2,3-dihydrobenzo[b][1,4]dioxin-6-amine (215 mg, 1.4 mmol) and triethylamine (170 mg, 1.7 mmol) in dichloromethane (10 ml). The reaction mixture was stirred at ambient temperature overnight, then concentrated. The crude product was purified by chromatography on silica gel (0-30% ethyl acetate in hexane) to give N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1,4-dimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulfonamida in the form of a white solid. MS (M+H, 404,10); 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,47 (s, 3H), 3,49 (s, 3H), of 4.12 (m, 4H), 6,55 (m, 1H), is 6.61 (d, J=2.4 Hz, 1H), 6,69 (d, J=8,4 Hz, 1H), 7,53 (m, 1H), to 7.59 (d, J=2 Hz, 1H), 10,00 (s, 1H).

IC50for the specified connection with the inhibition of bitter taste receptor was hT2R14 7,71 microns.

Example 6-3a: 1,4-dimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulphonylchloride

Chlorosulfonic acid (3 ml) was heated to 65ºC was added in portions 1,4-dimethylquinoxaline-2,3(1H,4H)-dione (PR who measures 6-3b, 1 g, 5.5 mmol) for 0.5 hour. The reaction mixture was stirred for 4 hours, then was cooled to ambient temperature and slowly poured into ice. The precipitate was filtered and washed with water to obtain 1,4-dimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulphonylchloride in the form of a white solid. 1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,50 (s, 6H), 7,25 (m, 2H), 7,38 (m, 4H).

Example 6-3b: 1,4-dimethylquinoxaline-2,3(1H,4H)-dione

To a solution of NaH (2.5 g) in DMF (200 ml) was added in portions cinoxacin-2,3(1H,4H)-dione (5 g)and then slowly added methyliodide (3.8 ml). The reaction mixture was stirred at ambient temperature for 4 hours, then added water (200 ml). The precipitate was collected by filtration and washed with water to obtain 1,4-dimethylquinoxaline-2,3(1H,4H)-dione as a white solid in 95% yield.1H NMR (400 MHz, DMSO-d6): δ (million parts: 3,50 (s, 6H), 7,25 (m, 2H), 7,38 (m, 4H).

Example 6-4: 1,4-dimethyl-2,3-dioxo-N-(4-(pyridine-2-ylmethyl)phenyl)-1,2,3,4-tetrahydroquinoxalin-6-sulfonamide

The compound is commercially available and was purchased through a company Ryan Scientific.

IC50for the specified connection with the inhibition of bitter taste receptor hT2R14 was 6,14 microns.

Additional connections are experimentally investigated and found the or, they have a relatively high level of effectiveness as inhibitors of the bitter taste receptor hT2R14. The results of this study are presented below in table D.

Table D
No. of connectionsConnectionhT2R14 IC50(µm)
6-5
N-(benzo[d][1,3]dioxol-5-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
0,264
6-6
N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
0,706
6-7
N-(3-fluoro-2-were)-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
0,935

6-8
N-(2,6-dimetilfenil)-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
6-9
1,3-dimethyl-2,4-dioxo-N-o-tolyl-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
cash consideration of USD 1,726
6-10
N-(4-forfinal)-1,4,7-trimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulfonamide
12,623
6-11
N-ethyl-N-m-tolyl-2-(N,1,4-trimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulfonamide)ndimethylacetamide
14,926
6-12
(Z)-3-(4-oxo-2-thioxo-5-(2,4,6-trimethoxyaniline)thiazolidin-3-yl)propanoic acid
9,371

6-13
(Z)-3-(5-(3,5-dimethoxybenzamide)-4-oxo-2-thioxothiazolidin-3-yl)propanoic acid
5,378
6-14
(E)-3-(benzo[d][1,3]dioxol-5-yl)-2-cyano-N-(5-phenyl-1,3,4-thiadiazole-2-yl)acrylamide
2,906
6-15
7-cyclopentyl-1,3-dimethyl-5-(2-ACS is propylthio)pyrimido[4,5-d]pyrimidine-2,4(1H,3H)-dione
5,428
6-16
N-cyclohexyl-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
7,239
6-17
N-(2-cyano-3, 5dimethylphenyl)-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
1,946

6-18
N-(3-fluoro-2-were)-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
0,935
6-19
N-(2,3-dimethylquinoxaline-6-yl)-1,4-dimethyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulfonamide
6,332
6-20
1,3-dimethyl-2,4-dioxo-N-o-tolyl-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
cash consideration of USD 1,726
6-22
N-ethyl-N-methyl-2-(2,6,8-trimethyl-5,7-dioxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidine-4-ylthio)ndimethylacetamide
0,861

6-23
N-(4-chloro-2-forfinal)-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydroquinazolin-6-sulfonamide
9,703
6-24
1,3,7-trimethyl-5-(2-oxo-2-(piperidine-1-yl)ethylthio)pyrimido[4,5-d]pyrimidine-2,4(1H,3H)-dione
0,292
6-25
1,3,7-trimethyl-5-(2-morpholino-2-oxometallic)pyrimido[4,5-d]pyrimidine-2,4(1H,3H)-dione
1,006
6-26
7-ethyl-1,3-dimethyl-5-(2-(4-methylpiperidin-1-yl)-2-oxometallic)pyrimido[4,5-d]pyrimidine-2,4(1H,3H)-dione
0,721

6-27
ethyl 4-(2-(2-ethyl-6,8-dimethyl-5,7-dioxo-5,6,7,8-tetrahydropyrimido[4,5-d]pyrimidine-4-ylthio)acetyl)piperazine-1-carboxylate
0,763
6-28
5-(2-(azepin-1-yl)-2-oxometallic)-1,3-dimethylpyrazol[2,3-d]pyrimidine-2,4(1H,3H)-dione
2,377
6-29
N-benzyl-2-(1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydropyrido[2,3-d]pyrimidine-5-ylthio)ndimethylacetamide
0,710
6-30
N-methyl-2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-6-sulfonamide
6,234

Table 7: Results of a representative screening for 23 receptor bitter taste

The compounds of examples 5 and 6, described above, were subjected to screening against a group of 23 of bitter taste receptors. The bitter taste receptors activated to EC80 corresponding agonist, then was treated with the above compounds at a concentration of 25 μm. The data summarized in the table below.

Inhibition of 80% or more=++++

Inhibition of 60%-80%=+++

Inhibition of 40%-60%=++

Inhibition of 20%-40%=+

Example 7. Data regarding the perception, for example, 10-10

To determine the effectiveness of individual antagonist conducted research regarding the taste using the specific agonist T2R8, the compounds and control blocker bitter taste. The authors of this and the finding was earlier proposed effective antagonist of hT2R8, which, as shown, has an effect on the taste, example 4-8 from provisional patent application U.S. serial No. 60/957129, filed August 21, 2007: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzo[d][1,3]dioxol-5-carboxamid. As shown, this substance reduces the bitter taste of coffee alone or in combination with blocker bitter taste a wide spectrum of action. As shown in table 6, when compared with the control antagonist according to the present invention (example 10-8) antagonist hT2R8 example 10-10 shows a greater ability to block the perception of bitter taste.

As has been shown in studies conducted in relation to taste in this example, the feeling of bitterness can be reduced or eliminated by introducing antagonists hT2R8, and the antagonist of example 10-10, apparently, is more potent analogue in comparison with the known antagonists bitter taste, such as N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)benzo[d][1,3]dioxol-5-carboxamid (7767). From this it follows that the sense of bitterness can be reduced or eliminated by introducing antagonists hT2R8 in compositions such as food, beverages and/or drugs, bitter taste which is caused by agonists T2R8.

Example 8. Identification of antagonists hT2R8

To identify antagonists of cell lines stably expressing hT2R8, together with promiscuous chimeric protein G16g44 were obtained as described in previous patent applications. High-performance analysis was carried out using stable cell lines and FLIPR (fluorometric tablet analyzer (Fluorescent Imaging Plate Reader)). Agonist hT2R8 used to activate receptors to 70-80% of their respective maximum activity. In the case of hT2R8 used agonist was Andrographolide (200 μm). To identify antagonists, together with the agonist was added compounds other chemical structures. Compounds that cause a statistically significant decrease in receptor activity, were pooled together and re-identify with a dose-dependent inhibition curves. The frame A and frame B were identified as antagonists of hT2R8 (figure 1). Specific examples are presented in table 1.

Example 9. Antagonists hT2R8

Example 9-1: 2-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)isoindoline-1,3-dione

2-(1H-pyrazole-4-yl)isoindoline-1,3-dione (example 9-1a) (1.5 g, 7 mmol), 4-(chloromethyl)for 3,5-dimethylisoxazol (1.5 g, 10 mmol) and cesium carbonate (3.3 g, 10 mmol) was stirred in DMF (20 ml) at 80ºC for 3 hours. The reaction mixture was cooled, diluted with H2O (150 ml) and extra is Aravali with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The solid product was ground into powder with ethyl acetate/hexane (1/9) and recrystallized from absolute ethanol (30 ml) by boiling under reflux to obtain 2-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)isoindoline-1,3-dione (900 mg, 38%) as a transparent, light-yellow solid.1H NMR (DMSO-d6, 400 MHz): δ of 1.36 (d, 3H), of 2.16 (s, 3H), 2,41 (s, 3H),J=7,2 Hz), with 5.22 (s, 2H), 7,81 (s, 1H), to $ 7.91-7,83 (m, 4H), 8,21 (s, 1H).MS M+H calculated 323,11;experimentally 323,1. Melting point: 170-171ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.18 micron.

Example 9-1a: 1-tosyl-1H-pyrazole-4-Amin

1-Tosyl-1H-pyrazole-4-amine (example 9-1b) (3 g, 12.7 mmol) and isobenzofuran-1,3-dione (1.9 g, 13 mmol) was stirred in DMF/acetonitrile (1/1) (20 ml) at 100°C for 1 hour. The mixture was cooled and diluted with H2O. the Precipitate was collected by filtration, washed with additional water, and then ethyl acetate and hexane. The solid product was dried in high vacuum to obtain 2-(1H-pyrazole-4-yl)isoindoline-1,3-dione (2.5 g, 92%) as a yellow solid. MS M+H calculated 214,1;experimentally 214,1.1H NMR (CDClsub> 3, 400 MHz): δ 7,93-8,10 (m, 6H), 13,03 (users, 1H).

Example 9-1b: 1-tosyl-1H-pyrazole-4-Amin

4-nitro-1-tosyl-1H-pyrazole (example 9-1c) (3 g, and 11.2 mmol) and 10% palladium-on-carbon (800 mg) in MeOH (150 ml) was stirred under 2 atmospheres of hydrogen hydrogenator Parra for 3 hours. The mixture was filtered through celite, concentrated and purified by chromatography on silica gel (80% ethyl acetate in hexane) to give 1-tosyl-1H-pyrazole-4-amine (1.9 g, 71%) as a pink solid.1H NMR (CDCl3, 400 MHz): δ 2.40 a (s, 3H), 3,01 (users, 2H), 7,29 (d, 2H,J=8 Hz), 7,41 (d, 1H,J=1.2 Hz), 7,53 (d, 1H,J=1.2 Hz), 7,81 (d, 2H,J=8 Hz).

Example 9-1c: 4-nitro-1-tosyl-1H-pyrazole

4-Nitro-1H-pyrazole (500 mg, 4.4 mmol), 4-methylbenzol-1-sulphonylchloride (840 mg, 4.4 mmol) and triethylamine (510 mg, 5 mmol) was stirred in DMF (25 ml) at 80°C for 1 hour. The reaction mixture was cooled, diluted with H2O (200 ml) and was extracted with ethyl acetate (3×, 100 ml). The organic phase is washed with aqueous 1N HCl (200 ml) and H2O (200 ml), dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The solid was ground into powder with hexane to obtain 4-nitro-1-tosyl-1H-pyrazole (600 mg, 50%) as off-white solid.1H NMR (DMSO-d6, 400 MHz): δ 2.40 a (s, 3H), 7,52 (d, 2H,J=8,4 Hz), 7,98 (d, 2H,J=8,8 Hz, to 8.57 (s, 1H), 9,58 (s, 1H).

Example 9-2: 2-((1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylamino)methyl)benzonitrile

1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (100 mg, 0.44 mmol), 2-(methyl bromide)benzonitrile (115 mg, 0.6 mmol) and triethylamine (0.5 ml, 3.5 mmol) in DMF (3 ml) was subjected to the action of radiation and stirred in a microwave reactor at 80ºC for 10 minutes. The reaction mixture was cooled, diluted with H2O (50 ml) and was extracted with ethyl acetate (3×30 ml). The combined organic extracts were dried over sodium sulfate and concentrated on a rotary evaporator. The residue was dissolved in ethanol (70 ml), H2O (3 ml) and acetic acid (1 ml) and the mixture is boiled under reflux for 2 hours. The solution was concentrated on a rotary evaporator, transferred in methanol (3 ml) and was purified by the method of reversed-phase HPLC, getting 3-1 ml aliquots (5-95% acetonitrile in H2O: a 16-minute gradient). Pure fractions were combined and concentrated on a rotary evaporator. The residue was ground into powder with ethyl acetate/hexane (1/9) to obtain 2-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)isoindoline-1-she (85 mg, 61%) as a pure white solid.1H NMR (DMSO-d6, 400 MHz): δ to 2.18 (s, 3H), 2,43 (s, 3H), equal to 4.97 (s, 2H), by 5.18 (s, 2H), 7,58-to 7.68 (m, 3H), to 7.77 (s, 1H), 8,10 (l,J=8 Hz, 1H), of 8.27 (s, 1H), 9,19 (users, 1H). As was the shows the above-mentioned compound inhibits bitter taste receptor hT2R08 and has a value IC50more than 30 μm.

Example 9-3: 2-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)isoindoline-1-he

2-((1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylamino)methyl)benzonitrile (example 9-2) (30 mg, 0.14 mmol) was stirred in a mixture of MeOH/2N aqueous NaOH (5 ml) at 100ºC for 30 minutes in a microwave reactor. The reaction mixture was acidified using 1N aqueous HCl (100 ml) and was extracted with ethyl acetate (3×, 70 ml). The combined organic extracts were dried over sodium sulfate and concentrated. The residue was dissolved in MeOH (3 ml) and was purified by the method of reversed-phase HPLC, getting 2-1,5 ml aliquots (5-95% acetonitrile in H2O: a 16-minute gradient). Pure fractions were combined and concentrated to obtain 2-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)isoindoline-1-it (21 mg, 50%) as pure white solid.1H NMR (DMSO-d6, 400 MHz): δ of 2.15 (s, 3H), 2,41 (s, 3H), to 4.81 (s, 2H), 5,17 (s, 2H), of 7.48-to 7.50 (m, 1H), 7,51-7,52 (m, 2H), 7,72 (l,J=7.2 Hz, 1H), of 7.75 (s, 1H), to 8.20 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC5011 microns.

Example 9-4: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)hinzelin-2,4(1H,3H)-dione

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine g is drochloride (75 mg, 0.33 mmol), methyl-2-isocyanatobenzene and triethylamine (200 mg, 2 mmol) in acetonitrile (3 ml) was exposed to radiation in a microwave reactor at 100ºC for 30 minutes. The reaction mixture was cooled, diluted with H2O (75 ml) and was extracted with ethyl acetate (3×50 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by chromatography on silica gel (75% ethyl acetate in hexane) to give 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)hinzelin-2,4(1H,3H)-dione (20 mg, 18%) as a light pink solid.1H NMR (DMSO-d6, 400 MHz): δ of 2.16 (s, 3H), 2.40 a (s, 3H), of 5.17 (s, 2H), 7,17-7,22 (m, 2H), 7,50 (s, 1H), 7,66 (dt,J=8, 1.2 Hz, 1H), 7,92 (l,J=6,8 Hz, 1H), 7,95 (s, 1H), to 11.52 (users, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501.5 μm.

Example 9-5: 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)tetrahydropyrimidin-2(1H)-he

1-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)tetrahydropyrimidin-2(1H)-he (example 9-5a) (50 mg, 0.18 mmol) and 60% sodium hydride (8 mg, 0.20 mmol) in DMF (3 ml) was stirred at room temperature for 15 minutes, then was cooled to 0ºC. To the mixture was added benzylbromide (31 mg, 0.18 mmol) and left to warm in the room for the Noah temperature, then was stirred for 2 hours. The reaction was suppressed with methanol and concentrated. The reaction mixture was diluted with brine (50 ml) and was extracted with dichloromethane (2×50 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated on a rotary evaporator. The residue was transferred to a dichloromethane (5 ml) and was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) to give 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)tetrahydropyrimidin-2(1H)-it (20 mg, 30%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 1.85 to 1.91 a (m, 2H), 2,10 (s, 3H), is 2.37 (s, 3H), of 3.12 (m, 2H), 3,55 (t,J=5.8 Hz, 2H), 4,48 (s, 2H), of 5.05 (s, 2H), 7,22-7,31 (m, 5H), 7,49 (s, 1H), to 7.84 (s, 1H). LC/MS; [M+H] calculated for C20H23N5O2; the expected value 366,19; experimentally 366,15. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 1.65 μm.

Example 9-5a: 1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)tetrahydropyrimidin-2(1H)-he

1-(3-Chlorpropyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea (example 9-5b) (200 mg, 0.64 mmol) and 60% sodium hydride (28 mg, 0.71 mmol) in DMF (2 ml) was stirred at 0ºC for 15 minutes, then left to warm to the room temperature and was stirred for 2 hours. The reaction was suppressed with methanol and concentrated. The reaction mixture was diluted with brine (50 ml) and was extracted with dichloromethane (2×50 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. The residue was transferred to a dichloromethane (5 ml) and was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) to give 1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)tetrahydropyrimidin-2(1H)-she (84 mg, 48%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 1.85 to 1.91 a (m, 2H), 2,10 (s, 3H), is 2.37 (s, 3H), of 3.12 (m, 2H), 3,48 (t,J=6,0 Hz, 2H), of 5.05 (s, 2H), to 6.57 (s, 1H), 7,46 (s, 1H), 7,80 (s, 1H). LC/MS; [M+H] calculated for C13H17N5O2; the expected value 276,14; experimentally 276,10.

Example 9-5b: 1-(3-chlorpropyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine (342 mg, 1.78 mmol) and 2-globabilization (213 mg, 1.78 mmol) in acetonitrile (5 ml) was heated at 65ºC for 16 hours. The reaction mixture was cooled to room temperature, concentrated and the residue was dissolved in dichloromethane (5 ml) and was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient). Pure fractions were combined was concentrically, then was ground into powder with ethyl acetate/hexane (1/9) and received (1-(3-chlorpropyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea (218 mg, 39%) as a white solid. LC/MS; [M+H] calculated for C12H16ClN5O2; the expected value 298,10; experimentally 298,10.

Example 9-6: 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2-he

1-Benzyl-5-(2,4-dimethoxybenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2-he (example 9-6a) (44 mg, 0.09 mmol), anisole (9 mg, 0.09 mmol) and 50% solution triperoxonane acid/dichloromethane (1 ml) in dichloromethane (1 ml) was stirred at room temperature for 2 hours. The reaction mixture was concentrated, reduce saturated sodium bicarbonate (50 ml), extracted with ethyl acetate (2×, 50 ml) and washed with brine (50 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. Purification using column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) allowed to obtain 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2-he (19 mg, 62%) as a white solid. LC/MS; [M+H] calculated for C19H22N6O2; the expected value 367,18; experimentally 67,20. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 5.84 microns.

Example 9-6a: 1-benzyl-5-(2,4-dimethoxybenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2-he

1-Benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea (example 9-6b) (50 mg, 0.15 mmol), 2,4-methoxybenzylamine (26 mg, 0.15 mmol) and formaldehyde (37% wt. in water) (25 mg, 0.31 mmol) was heated at 100ºC for 16 hours. The reaction mixture was cooled to room temperature, diluted with brine (50 ml) and was extracted with dichloromethane (2×50 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) to give 1-benzyl-5-(2,4-dimethoxybenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2-she (44 mg, 56%) as oil. LC/MS; [M+H] calculated for C28H32N6O4; the expected value 517,25; experimentally 517,20.

Example 9-6b: 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine (287 mg, 1,49 mmol) and basilidians (199 mg, 1,49 mmol) in acetonitrile (5 ml) on revali at 65ºC for 16 hours. The reaction mixture was cooled to room temperature and concentrated. The residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) and was ground into powder with ethyl acetate/hexane (1/9) to obtain 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea (246 mg, 51%) as a white solid. LC/MS; [M+H] calculated for C17H19N5O2; the expected value 326,15; experimentally 326,10.

Example 9-7: 5-benzyl-1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethyl-1,3,5-triazine-2-he

Was obtained as in example 9-6a 1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethylamine (example 9-7a), formaldehyde and benzylamine. Yield: 15%.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), is 2.37 (s, 3H), 2,69 (t,J=7,6 Hz, 2H), 3,39 (t,J=7.8 Hz, 2H), 3,81 (s, 2H), 4.16 the (s, 2H), 4,42 (s, 2H), of 5.05 (s, 2H), 7,17-to 7.32 (m, 10H), 7,42 (s, 1H), 7,78 (s, 1H). LC/MS; [M+H] calculated for C27H30N6O2; the expected value 471,24; experimentally 471,15. As shown, the above-mentioned compound inhibits bitter receptor hT2R08 and has the IC50of 2.64 mm.

Example 9-7a: 1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethylamine

Was obtained as in example 9-6b 1-((3,5-dimethylisoxazol-4-yl)METI is)-1 H-pyrazole-4-amine and geneticization. Yield: 29%.1H NMR (DMSO-d6, 400 MHz): δ 2,10 (s, 3H), of 2.36 (s, 3H), 2,69 (t,J=7.2 Hz, 2H), 3,25 (kV,J=7,4 Hz, 2H), 5,02 (s, 2H), 6,00 (t,J=5.8 Hz, 1H), 7,16-7,30 (m, 6H), 7,68 (s, 1H), 8,13 (s, 1H). LC/MS; [M+H] calculated for C18H21N5O2; the expected value 340,17; experimentally 340,20.

Example 9-8: 1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethyl-1,3,5-triazine-2,4,6-Trion

1-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethylamine (example 9-7a) (100 mg, 0.29 mmol) in THF (2 ml) was cooled to 0ºC and was slowly added n-chlorocarbonylsulfenyl (93 mg, 0.88 mmol). After the addition the reaction mixture was left to warm to room temperature and was stirred for 1 hour. The reaction mixture was concentrated and the residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) to give (1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethyl-1,3,5-triazine-2,4,6-trione (100 mg, 83%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2.13 in (s, 3H), 2,39 (s, 3H), 2,82 (t,J=5.8 Hz, 2H), 3,88 (t,J=8.0 Hz, 2H), 5,17 (s, 2H), 7,19-7,31 (m, 5H), 7,44 (s, 1H), 7,89 (s, 1H), 11,84 (s, 1H). LC/MS; [M+H] calculated for C20H20N6O4; the expected value 409,15; experimentally 409,20. As shown, Sanasanne compound inhibits bitter taste receptor hT2R08 and has the IC 503,03 microns.

Example 9-9: 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2,4,6-Trion

1-Benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea (example 95b) (100 mg, 0.31 mmol) in THF (2 ml) was cooled to 0ºC and was slowly added n-chlorocarbonylsulfenyl (97 mg, of 0.92 mmol). After the addition the reaction mixture was left to warm to room temperature and was stirred for 1 hour. The reaction mixture was concentrated and the residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) to give 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2,4,6-trione (112 mg, 93%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2,12 (s, 3H), of 2.38(s, 3H), to 4.87 (s, 2H), 5,16 (s, 2H), 7.23 percent-7,34 (m, 5H), was 7.45 (s, 1H), of 7.90 (s, 1H), 11,93 (s, 1H). LC/MS; [M+H] calculated for C19H18N6O4; the expected value 395,14; experimentally 395,15. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501,14 mm.

Example 10. Antagonists hT2R8: obtain the compounds according to the invention

Example 10-1: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-shall irsol-4-carbonylated (example 10-1a) (6 g, 25.5 mmol) in toluene (100 ml) was boiled under reflux for 1 hour and cooled to ambient temperature in a nitrogen atmosphere. Added the hydrochloride of the methyl ester of glycine (3.1 g, 26 mmol) and triethylamine (3.2 g, 32 mmol), the mixture was boiled under reflux for 16 hours. The reaction mixture was cooled and solvent was removed on a rotary evaporator. The solid was re-dissolved in ethyl acetate (100 ml) and the organic phase is washed with 1N HCl solution (2×150 ml). The aqueous phase was again extracted with ethyl acetate (2×, 75 ml)and the combined organic extracts were dried over sodium sulfate and concentrated. The resulting solid was ground into powder with ethyl acetate/hexane (1/9), and dried under high vacuum to obtain 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (5,2 g, 74%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2,19 (s, 3H), 2,42 (s, 3H), 4.09 to (s, 2H), is 5.06 (s, 2H), 5,68 (users, 1H), of 7.90 (s, 1H), 8,05 (1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501,7 mm.

Example 10-1a: 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated

Sodium nitrite (450 mg, 6.5 mmol, H2O) (10 ml) was added dropwise during 10 minutes to a suspension of 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-p is razol-4-carbohydrazide (example 10-1b) (1 g, 4.3 mmol) in 10% aqueous acetic acid (50 ml) and was cooled to 0ºC with a bath of ice water. The reaction mixture was stirred for additional 15 minutes, then was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts washed with aqueous saturated sodium carbonate (100 ml), and then H2O (100 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The solid product was ground into powder with ethyl acetate/hexane (1/9) and dried under vacuum to obtain 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (1 g, 93%) as a white solid.1H NMR (CDCl3, 400 MHz): δ of 2.20 (s, 3H), of 2.44 (s, 3H), 5,07 (s, 2H), 7,81 (s, 1H), to 7.93 (s, 1H).

Example 10-1b: 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbohydrazide

Ethyl-1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carboxylate (example 10-1c) (6 g, 24 mmol) and hydrazine (7.5 g, 240 mmol) was stirred in EtOH (100 ml) by boiling under reflux for 12 hours. The solution was concentrated on a rotary evaporator and the solid product was ground into powder with ethyl acetate/hexane (1/9), and dried under high vacuum to obtain 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbohydrazide (5.5 g, 97%) as a pure white solid.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), 2,39 (s, 3), further 5.15 (s, 2H), 7,81 (s, 1H), 8,17 (s, 1H), 9,31 (users, 1H).

Example 10-1c: 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carboxylate

Ethyl-1H-pyrazole-4-carboxylate (4,2 g, 30 mmol), 4-(chloromethyl)for 3,5-dimethylisoxazol (5,1 g, 35 mmol) and cesium carbonate (9.8 g, 30 mmol)in DMF (50 ml)was stirred at 80ºC for 12 hours. The reaction mixture was cooled to ambient temperature, diluted with 0.1 N HCl (150 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate and concentrated on a rotary evaporator. The solid product was ground into powder with ethyl acetate/hexane (1/9) and collected by filtration to obtain ethyl-1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carboxylate (6 g, 80%) as a white solid.1H NMR (CDCl3, 400 MHz): δ of 1.34 (t,J=7.2 Hz, 3H), 2,19 (s, 3H), 2,43 (s, 3H), 4,29 (kV,J=7.2 Hz, 2H), is 5.06 (s, 2H), to 7.77 (s, 1H), to $ 7.91 (s, 1H).

Example 10-2: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-1 of the hydrochloride metilenovogo ether (+/-)-phenylalanine and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated. Yield: 58%.1H NMR (acetone-d6, 400 MHz): δ 2,17 (s, 3H), 2,43 (s, 3H), of 3.07 (DD,J=14.4V, 6.4 Hz, 1H), 3,20 (DD,J=14, and 4.4 Hz, 1H), 4.53-in (t,J=4,8 Hz, 1H), 5,18 (s, 2H), 7,27-7,19 (m, 5H), 7,46 (users, 1H), 7,79 (s, 1H, to 7.99 (s, 1H). MS M+H calculated 366,15; experimentally 366,1. Melting point: 169-171ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.18 micron.

Example 10-3: (S)-5-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-1 of methyl ester hydrochloride (S)-phenylalanine and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 6a). Yield: 13% allocated using chiral chromatography.1H NMR (CDCl3, 400 MHz): δ 2,19 (s, 3H), 2,42 (s, 3H), 2,88 (DD,J=13,6, and 9.2 Hz, 1H), 3,35 (DD,J=13,6, 3.6 Hz, 1H), or 4.31 is 4.35 (m, 1H) of 5.06 (s, 2H), of 5.53 (users, 1H), 7,21 and 7.36 (m, 5H), the 7.85 (s, 1H), 8,01 (s, 1H). LC/MS; [M+H] calculated for C19H19N5O3; the expected value 366,15; experimentally 366,1. [α]D=(is)-136, c=0,1, ethanol. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,12 ám.

Example 10-4: (R)-5-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-1 of methyl ester hydrochloride (R)-phenylalanine and 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated. Yield: 9% allocated using chiral chromatography1H NMR (CDCl3, 400 MHz): δ 2,19 (s, 3H), 2,42 (s, 3H), 2,88 (DD,J=13,6, 9,2 is C, 1H), 3,35 (DD,J=13,6, 3.6 Hz, 1H), or 4.31 is 4.35 (m, 1H) of 5.06 (s, 2H), of 5.53 (users, 1H), 7,21 and 7.36 (m, 5H), the 7.85 (s, 1H), 8,01 (s, 1H). MS M+H calculated 366,15; experimentally 366,1. [α]D=(+)-124, c=0,2, ethanol. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.11 μm.

Example 10-5: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-phenoxyethyl)imidazolidin-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) (200 mg, 0.7 mmol), (2 bromoethoxy)benzene (200 mg, 1 mmol) and cesium carbonate (325 mg, 1 mmol) was subjected to the action of radiation in a microwave reactor at 85ºC for 20 minutes. The reaction mixture was cooled to room temperature, diluted with aqueous 1N HCl (100 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The residue was transferred to a methanol (10 ml) and was purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: 25 minute gradient). Pure fractions were combined, concentrated, and then re-dissolved in absolute ethanol and concentrated on a rotary evaporator (4×) to give 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-phenoxyethyl)imidazolidin-2,4-dione (150 mg, 54%) as a white solid.1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H, to 2.41 (s, 3H), 3,86 (t,J=5,2 Hz, 2H), 4,19 (t,J=4.4 Hz, 2H), 4,25 (s, 2H), of 5.05 (s, 2H), to 6.88 (DD,J=9,2, 1.2 Hz, 2H), 7,00 (dt,J=to 7.6, 1.2 Hz, 1H), 7,27-to 7.32 (m, 2H), 7,89 (s, 1H), with 8.05 (s, 1H). MS M+H calculated 396,17; experimentally 396,1. Melting point: 117-118ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-6: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-methoxybenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 6) and 3-methoxybenzylamine. Yield: 55%.1H NMR (CDCl3, 400 MHz): δ 2,19 (s, 3H), 2,42 (s, 3H), 3,81 (s, 3H), 3,85 (s, 2H), 4,58 (s, 2H), is 5.06 (s, 2H), for 6.81-to 6.88 (m, 3H), 7,26-7,31 (m, 1H), 7,92 (s, 1H), 8,08 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.07 μm.

Example 10-7: Methyl 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzoate

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione and 3-methoxybenzylamine. Yield: 83%.1H NMR (CDCl3, 400 MHz): δ of 2.20 (s, 3H), 2,43 (s, 3H), 3,86 (s, 2H), 3,93 (s, 3H), of 4.67 (s, 2H), 5,07 (s, 2H), 7,45-7,52 (m, 2H), to 7.93 (s, 1H), 7,95 (s, 1H), 8,03 (DD,J=to 7.2, 1.6 Hz, 1H), 8,08 (s, 1H). As shown, the above-mentioned connection inhibi is the duty to regulate the bitter taste receptor hT2R08 and has the IC 500,09 ám.

Example 10-8: 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzoic acid

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (500 mg, 1.8 mmol) (example 10-5), methyl-3-(methyl bromide)benzoate (456 mg, 2 mmol) and cesium carbonate (650 mg, 2 mmol) was stirred in DMF (4 ml) in a microwave reactor at 85°C for 20 minutes. The reaction mixture was cooled, diluted with 1N aqueous HCl (100 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The crude ester was dissolved in methanol (5 ml) was added aqueous NaOH (50 ml, 10% wt.) and the mixture was stirred at ambient temperature for 2 hours. The reaction mixture was acidified using 1N HCl (150 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated on a rotary evaporator. The free acid was ground into powder with ethyl acetate/hexane (1/9) and dried under vacuum to obtain 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzoic acid (610. mg, 83%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2.13 in (s, 3H), to 2.29 (s, 3H), 4.00 points (s, 2H), 4,59 (s, 2H), by 5.18 (s, 2H), 7,46-to 7.59 (m, 2H), 7,78 (s, 1H), 7,85-7,88(m, 2H), 8,18 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 1.8 microns.

Example 10-9: 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)-N-methylbenzamide

3-((3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzoic acid (100 mg, 0.24 mmol) (example 10-8), methylamine hydrochloride (67 mg, 1 mmol), triethylamine (155 mg, 1.5 mmol) and EDC (57 mg, 0.3 mmol) in acetonitrile (3 ml) was exposed to radiation in a microwave reactor at 80ºC for 10 minutes. The reaction mixture was cooled, diluted with aqueous 1N HCl (100 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The crude product was dissolved in MeOH (3 ml) and was purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: 25 minute gradient) to give 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)-N-methylbenzamide (25 mg, 25%) as a white solid. MS M+H calculated 423,17; experimentally 423,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,14 m.

Example 10-10: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-hydroxybenzyl them who solidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-(3-hydroxybenzylidene)acetate (example 10-10a). Yield: 24%.1H NMR (DMSO, 400 MHz): δ of 2.15 (s, 3H), 2,41 (s, 3H), 3,99 (s, 2H), of 4.45 (s, 2H), total of 5.21 (s, 2H), 6,70 (m, 3H), 7,15 (m, H), 7,80 (s, 1H), 8,19 (s, H), 9,44 (s, H). LC/MS; [M+H] expected value 382,1; experimentally 382,1. Melting point: 35-136ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50a 0.035 microns.

Example 10-10a: methyl-2-(3-hydroxybenzylidene)acetate

Methyl ester of glycine (500 mg, 4 mmol) and 3-hydroxybenzaldehyde (480 mg, 4 mmol) was dissolved in 5 ml of a mixture of THF/methanol (1:1). In the reaction mixture was slowly added acetic acid (240 mg, 4 mmol) and 1M Lamborghini sodium in THF (4.8 ml, 4.8 mmol). The reaction mixture was subjected to irradiation in a microwave reactor at 85ºC for 15 minutes, cooled to room temperature and removing the salt by filtration. The clear solution was concentrated and the residue was purified by the method of reversed-phase HPLC (10-95% acetonitrile in H2O: 25 minute gradient) to give the above compound in the form of a transparent gel. Yield 45%. MS M+H calculated 196,1; experimentally 196,1.

Example 10-11: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-piraso the-4-yl)-1-(2-hydroxybenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-(2-hydroxyethylamino)acetate (example 10-11a). Yield: 28%.1H NMR (DMSO, 400 MHz): δ 2,12 (s, 3H), of 2.38 (s, 3H), 4.00 points (s, 2H), of 4.45 (s, 2H), 5,17 (s, 2H), 6,83 (m, 2H), 7,10 (m, 2H), 7,78 (s, 1H), 8,16 (s, H), to 9.66 (s, H). LC/MS; [M+H] expected value 382,1; experimentally 382,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.07 μm.

Example 10-11a: methyl-2-(2-hydroxyethylamino)acetate

Was obtained as in example 10-10a of the methyl ester of glycine and 2-hydroxybenzaldehyde. Yield 40%. MS M+H calculated 196,1; experimentally 196,1

Example 10-12: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-hydroxybenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated and methyl-2-(4-hydroxybenzylidene)acetate (example 10-12a). Exit 9%.1H NMR (DMSO, 400 MHz): δ 2,117 (s, 3H), 2,383 (s, 3H), 3,918 (s, 2H), 4,387 (s, 2H), 5,174 (s, 2H), 6,719 (J=8,8, d, 2H), 7,108 (J=8,8 m, 2H), 7,761 (s, 1H), 8,154 (s, H)9,399 (s, H). LC/MS; [M+H] expected value 382,1; experimentally 382,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-12a: methyl-2-(4-hydroxyben is ylamino)acetate

Was obtained as in example 10-10a of the methyl ester of glycine and 4-hydroxybenzaldehyde. Yield 40%. MS M+H calculated 196,1; experimentally 196,1

Example 10-13: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-hydroxy-4-methoxybenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-(3-hydroxy-4-methoxybenzylamine)acetate (example 10-13a). Yield 22%.1H NMR (DMSO, 400 MHz): δ 2,119 (s, 3H), 2,383 (s, 3H), 3,716 (s, 3H), 3,923 (s, 2H), 4,361 (s, 2H), 5,117 (s, 2H), 6,667 (m, 2H), 6,863 (J=8,4, d, 1H), 7,766 (s, H)8,159 (s, H). MS M+H calculated 412,1; experimentally 412,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.1 ám.

Example 10-13a: methyl-2-(3-hydroxy-4-methoxybenzylamine)acetate

Was obtained as in example 10-10a of the methyl ester of glycine and 3-hydroxy-4-methoxybenzaldehyde. Yield 47%. MS M+H calculated 226,1; experimentally 226,1

Example 10-14: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-(2-methoxyethoxy)benzyl)imidazolidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-(3-(2-methoxyethoxy)benzylamino)acetate (example 0-14a). The output is 27%.1H NMR (DMSO, 400 MHz): δ 2,12 (s, 3H), of 2.38 (s, 3H), 3,26 (s, 3H), 3,62 (t, J=4,4, 2H), 3,98 (s, 2H), 4,06 (t, J=4,4, 2H), 4,48 (s, 2H), by 5.18 (s, 2H), 6,86 (m, 3H), 7,24 (t, J=8, 1H), 7,78 (s, 1H), 8.17 and (s, 1H). MS M+H calculated 440,2; experimentally 440,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.07 μm.

Example 10-14a: methyl-2-(3-(2-methoxyethoxy)benzylamino)acetate

Was obtained as in example 10-10a of the methyl ester of glycine and 3-(2-methoxyethoxy)benzaldehyde. Yield 55%. MS M+H calculated 254,1; experimentally 254,1

Example 10-15: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(methylthio)benzyl)imidazolidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-(2-(methylthio)benzylamino)acetate (example 10-15a). Yield 67%.1H NMR (DMSO, 400 MHz): δ 2,12 (s, 3H), 2,39 (s, 3H), 2,48 (s, 3H), 3,98 (s, 2H), of 4.54 (s, 2H), 5,19 (s, 2H), 7,18 (m, 1H), 7,30 (m, 3H), 7,79 (s, 1H), 8,18 (s, 1H). LC/MS; [M+H] expected value 412,1; experimentally 412,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.03 micrometers.

Example 10-15a: methyl-2-(2-(methylthio)benzylamino)acetate

Was obtained as in example 10-10a of the methyl ester of glycine and 2-(methylthio)benzaldehyde. The yield is 50%. MS M+H RAS is Chicano 226,1; experimentally 226,1

Example 10-16: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(2-methoxyethoxy)benzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-hydroxybenzyl)imidazolidin-2,4-dione (example 10-11) and 2-pomatoleios ether. Exit 19%.1H NMR (DMSO, 400 MHz): δ 2,11 (s, 3H), of 2.08 (s, 3H), of 3.25 (s, 3H), of 3.64 (t, J=3,6, 2H), 4.00 points (s, 2H), 4,11 (t, J=3,2, 2H), 4,27 (s, 2H), 5,17 (s, 2H), 6.90 to (m, 1H), 7,00 (m, 1H), 7,26 (m, 2H), 7,76 (s, 1H), of 8.15 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.1 ám.

Example 10-17: 33-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(methylsulfinyl)benzyl)imidazolidin-2,4-dione

A 20-ml vessel for use in the microwave system was dissolved 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(methylthio)benzyl)imidazolidin-2,4-dione (example 10-15) (70 mg, 0,17 mmol) and m-HPBC (m-chloroperbenzoic acid) (58 mg, 0.34 mmol) in dichloromethane at 0ºC. The reaction mixture was stirred at 0ºC and left to warm to room temperature for 4 hours. The reaction solvent was removed under vacuum, and the crude product was dissolved in 1 ml ethanol and purified by HPLC Varian (10-95% acetonitrile/water; 25 minutes). Purified fractions were evaporated under vacuum to obtain to enter the named connection. MS M+H calculated 428,1; experimentally 428,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.4 µm. Yield: 12 mg, 17%.

Example 10-18: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-methoxybenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(methyl bromide)-2-methoxybenzene (199 mg, 1 mmol). Yield: 33%. MS M+H calculated 396,1; experimentally 396,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-19: 2-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzonitrile

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 2-(methyl bromide)benzonitrile. Yield: 27%. MS M+H calculated to € 391.1; experimentally to € 391.1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.5 micron.

Example 10-20: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-methylbenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dio is a (example 10-1) and 1-(methyl bromide)-2-methylbenzene. Yield: 21%. MS M+H calculated 380,1; experimentally 380,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.1 ám.

Example 10-21: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-terbisil)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(methyl bromide)-2-fervently. Yield 42%. MS M+H calculated 384,1; experimentally 384,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.08 μm.

Example 10-22: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(trifluoromethyl)benzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(methyl bromide)-2-(trifluoromethyl)benzene. Yield: 37%. MS M+H calculated of 434.1; of 434.1 experimentally. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.2 μm.

Example 10-23:3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-nitrobenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) is 1-(methyl bromide)-2-nitrobenzene. Yield 22%. MS M+H calculated 411,1; experimentally 411,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.07 μm.

Example 10-24: 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzaldehyde

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3-(methyl bromide)benzaldehyde. Yield: 35%.1H NMR (DMSO, 400 MHz): δ 2,123 (s, 3H), 2,388 (s, 3H), 4,035 (s, 2H), 4,631 (s, 2H), 5,186 as (s, 2H), 7,581 (m, 1H), 7,643 (m, 1H), 7,787 (m, 3H), 8,178 (s, H)9,997 (s, H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.2 μm.

Example 10-25: 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzonitrile

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3-(methyl bromide)benzonitrile. The output of 21%. MS M+H calculated 411,1; experimentally 411,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501 micron.

Example 10-26: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-methylbenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dime isoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(methyl bromide)-3-methylbenzene. Yield 25%. MS M+H calculated 380,1; experimentally 380,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.02 mm.

Example 10-27: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-terbisil)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(methyl bromide)-3-fervently. The output is 27%. MS M+H calculated 384,1; experimentally 384,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-28: 1-((1,5-dimethyl-1H-pyrazole-3-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3-(methyl bromide)-1,5-dimethyl-1H-pyrazole. Yield: 22%. MS M+H calculated 384,1; experimentally 384,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,3 ám

Example 10-29: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-methoxybenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 0-1) and 1-(methyl bromide)-4-methoxybenzene. Exit 19%. MS M+H calculated 396,1; experimentally 396,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.07 μm.

Example 10-30: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-methylbenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(methyl bromide)-4-methylbenzene. Yield 25%. MS M+H calculated 380,1; experimentally 380,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-31: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-terbisil)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(methyl bromide)-4-fervently. Exit 33%. MS M+H calculated 384,1; experimentally 384,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.05 microns.

Example 10-32: 4-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzonitrile

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 4-(bromet the l)benzonitrile. The output of 21%. MS M+H calculated to € 391.1; experimentally 391,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.05 microns.

Example 10-33: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-(hydroxymethyl)benzyl)imidazolidin-2,4-dione

3-((3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzaldehyde (example 10-24) (131 mg, 0.3 mmol) was dissolved in 2 ml ethanol. The solution was passed through the apparatus of the H-Cube at room temperature using 10% Pd/C catalyst at a flow rate of 1 ml/min. The collected fraction was concentrated, re-dissolved in 2 ml ethanol and purified by HPLC (10-95% acetonitrile/water, 25 minutes). Purified fractions were combined and concentrated to obtain the above-mentioned compounds.1H NMR (DMSO, 400 MHz): δ 2,123 (s, 3H), 2,388 (s, 3H), 3,978 (s, 2H), 4,516 (s, 2H), at 5,182 (s, 2H), 7,242 (m, 4H), 7,779 (s, 1H), 8,172 (s, 1H), 8,505 (s, 1H). MS M+H calculated 396,1; experimentally 396,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.3 microns. Yield: 24 mg, 18%.

Example 10-34: 1-(2-aminobenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-nitrobenzyl)imidazolidin-2,4-dione (example 10-23) (126 mg,0.3 mmol) was dissolved in 2 ml ethanol. The solution was passed through the apparatus of the H-Cube at room temperature using 10% Pd/C catalyst at a flow rate of 1 ml/min. The collected fraction was concentrated, re-dissolved in 2 ml ethanol and purified by HPLC (10-95% acetonitrile/water, 25 minutes). Purified fractions were combined and concentrated to obtain the above-mentioned compounds. Yield 26%. MS M+H calculated 381,1; experimentally 381,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.02 mm.

Example 10-35:1-(3,4-dimethoxybenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) (275 mg, 1 mmol), (3,4-acid)methanol (201 mg, 1.2 mmol), N,N,N,N-tetramethyldisiloxane (344 mg, 2 mmol) was dissolved in 2 ml anhydrous THF. Added tributylphosphine (404 mg, 2 mmol) and the reaction mixture is placed in a microwave reactor for 5 min at 90ºC. The reaction mixture was filtered, concentrated and purified by HPLC (10-95% acetonitrile/water, 25 minutes) to obtain the above-mentioned compounds. Yield: 25 mg, 6%.1H NMR (DMSO, 400 MHz): δ 2,119 (s, 3H), 2,385 (s, 3H), 3,724 (J=6,4d, 6H), 3,946 (s, 2H), 4,435 (s, 2H), 5,178 (s, 2H), 6,885 (m, 3H), 7,776 (s, 1H), 8,166 (s, 1H). MS M+H calculated 426,1; experimentally 426,1. As shown, enter the named compound inhibits bitter taste receptor hT2R08 and has the IC 500.06 micron.

Example 10-36: 1-(benzo[d][1,3]dioxol-5-ylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-35 of 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and benzo[d][1,3]dioxol-5-ylmethanol. Yield: 19%.1H NMR (DMSO, 400 MHz): δ 2,143 (s, 3H), 2,408 (s, 3H), 3,977 (s, 2H), 4,440 (s, 2H), 5,202 (s, 2H), 6,003 (s, 2H), 6,897 (m, 3H), 7,788 (s, 1H), 8,181 (s, 1H). MS M+H calculated 410,1; experimentally 410,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.07 μm.

Example 10-37: 1-(3-((dimethylamino)methyl)benzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) (275 mg, 1 mmol), 1,3-bis(methyl bromide)benzene (263 mg, 1 mmol) and cesium carbonate (325 mg, 1 mmol) was dissolved in 2 ml DMF and was irradiated in a microwave reactor at 165ºC for 5 minutes. The reaction mixture was cooled to room temperature and removed sediment in the form of salts by filtration. A clear solution containing the crude product was concentrated and re-dissolved in ethyl acetate. The organic solution was twice washed with water and then brine. The organic phase was dried over sodium sulfate is evaporated to obtain the crude product, which is transferred to the next stage without further purification or analysis. 1-(3-(methyl bromide)benzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 42a) (152 mg, 0.3 mmol), dimethylamine (2 M solution in THF) (1.5 ml, 3 mmol) and sodium hydride (9 mg, 0.36 mmol) was dissolved in 1 ml anhydrous THF. The reaction mixture was placed in a microwave reactor for 5 min at 120ºC. The crude product was re-dissolved in 2 ml ethanol and purified by HPLC (10-95% acetonitrile/water, 25 minutes) to give 1-(3-((dimethylamino)methyl)benzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (16 mg, 13%). MS M+H calculated 423,1; experimentally 423,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501.2 microns.

Example 10-38: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((1-methyl-1H-pyrazole-3-yl)methyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3-(methyl bromide)-1-methyl-1H-pyrazole. Exit 19%. MS M+H calculated 370,1; experimentally 370. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.4 µm.

Example 10-39: N-(2-((3,5-dimethylisoxazol-4-yl)methyl)-2H-tetrazol-5-yl)-3-methoxybenzamide

2-((3,5-Dimethylisoxazol-4-yl)methyl)-2H-tetrazol-5-amine (example 10-39a) (102 mg, 0,528 mmol), 3-methoxybenzoyl chloride (0,065 ml, 0,528 mmol) and pyridine (0,043 ml, 0,528 mmol) in acetonitrile (3 ml) was stirred at 100ºC for one hour. The reaction mixture was diluted with dichloromethane (30 ml) and washed with brine (30 ml). Organic matter was dried over sodium sulfate, concentrated and purified by the method of reversed-phase HPLC (solvent system: acetonitrile/water, 10%-100% gradient, 25-minute series), obtaining N-(2-((3,5-dimethylisoxazol-4-yl)methyl)-2H-tetrazol-5-yl)-3-methoxybenzamide in the form of a white crystalline solid (60 mg, 35% yield) MS M+H calculated 329,1, experimentally 329. 1H NMR (400 MHz, DMSO-d6): δ 2,02 (s, 3H), of 2.46 (s, 3H), 3,81 (s, 3H), 5,78 (s, 2H), 7,16 (m, 1H), 7,42 (t, J=8 Hz, 2H), 7,54 (m, 1H), and 11.3 (s, 1H). IC50for the specified connection with the inhibition of bitter taste receptor hT2R8 amounted to 1.87 μm.

Example 10-39a: 2-((3,5-dimethylisoxazol-4-yl)methyl)-2H-tetrazol-5-amine

2H-tetrazol-5-amine (1.29 g, 12.5 mmol), 4-(chloromethyl)for 3,5-dimethylisoxazol (1,56 ml, 12.5 mmol) and potassium carbonate (1.73 g, a 15.5 mmol) in DMF (20 ml) was heated to 80ºC under stirring for 16 hours. The reaction mixture was cooled to room temperature, diluted with dichloromethane (100 ml) and washed sequentially with saline and water. Organic matter is dried over sodium sulfate and concentrated using a rotary evaporator. The crude product was purified by chromatography on silica gel (0-10% gradient ethyl acetate/dichloromethane)to give 2-((3,5-dimethylisoxazol-4-yl)methyl)-2H-tetrazol-5-amine as a white crystalline solid (970 mg, 40% yield). MS M+H calculated 195,1, experimentally 195.

Example 10-40: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-imidazol-4-yl)benzo[d][1,3]dioxol-5-carboxamid

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-imidazol-4-amine (example 10-40a) (110 mg, or 0.57 mmol), benzo[d][1,3]dioxol-5-carbonyl chloride (105 mg, or 0.57 mmol) and triethylamine (90 μl, 0.69 mmol) in dichloromethane was stirred for 16 hours. The reaction mixture was diluted with dichloromethane (30 ml) and washed sequentially with saline and water. Organic matter was dried over sodium sulfate and concentrated by rotary evaporation. The crude product was purified by the method of reversed-phase HPLC (solvent system: acetonitrile/water, 10%-100% gradient, 25-minute series) to obtain N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-imidazol-4-yl)benzo[d][1,3]dioxol-5-carboxamide as a white crystalline solid (32 mg, 15% yield). MS M+H calculated 341,3 experimentally 341,3. 1H NMR (400 MHz, DMSO-d6): δ of 2.08 (s, 3H), 2,41 (s, 3H), 5,02 (s, 2H), 6,07 (s, 2H), 6,95 (d, J=8,4 Hz, 1H), 7,27 (d, J=1.6 Hz, 1H), 7,54 (m, 3H), or 10.6 (s, 1H). IC50for the specified connection with the inhibition of re is of aptara bitter taste hT2R8 was 12.1 microns.

Example 10-40a: 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-imidazol-4-amine

3,5-Dimethyl-4-((4-nitro-1H-imidazol-1-yl)methyl)isoxazol (example 10-40b) (1.0 g, 4.5 mmol) and 10% palladium-on-charcoal (200 mg) in methanol (40 ml) was shaken in apparatus for shaking Parra at a pressure of 2.5 bar hydrogen for 2 hours. After filtration through a layer of celite followed by rotary evaporation was obtained 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-imidazol-4-amine in the form of a yellowish-red solid (800 mg, 93% yield). MS M+H calculated 193, experimentally 193.

Example 10-40b: 3,5-dimethyl-4-((4-nitro-1H-imidazol-1-yl)methyl)isoxazol

3,5-Dimethyl-4-((4-nitro-1H-imidazol-1-yl)methyl)isoxazol got a way similar to that described in example 10-41c, by alkylation of 4-nitro-1H-imidazole, getting 3,5-dimethyl-4-((4-nitro-1H-imidazol-1-yl)methyl)isoxazol in the form of a white crystalline solid (5.0 g, 80% yield). MS M+H calculated 223, experimentally 223. 1H NMR (400 MHz, DMSO-d6): δ (million parts of 2.09 (s, 3H), 2,43 (s, 3H), of 5.15 (s, 2H), of 7.90 (d, J=1.6 Hz, 1H), 8,35 (d, J=1.9 Hz, 1H).

Example 10-41: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylimidazolidine-2,4-dione

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 10-41a) (1.0 g, 5,20 mmol), ethyl-2-sozialprojekte (0,745 g, 5,20 mmol) and triethylamine (1.5 ml, 10.4 mmol) were mixed in EtO (20 ml). The reaction mixture is boiled under reflux for 12 hours and then left to cool to room temperature. The solvent was removed under vacuum, after some time formed crystals. The crystals were collected and washed with hexane to obtain 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylimidazolidine-2,4-dione in 80% yield as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 1,53-is 1.51 (d, 3H), 2,19 (s, 3H), 2,42 (s, 3H), 4,21-4,19 (m, 1H), is 5.06 (s, 2H), 6,00 (users, 1H), of 7.90 (s, 1H), with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 1.3 μm.

Example 10-41a: 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride

Tert-butyl-1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylcarbamate (example 10-41b) (592 mg, 2 mmol) was stirred in a solution of 4N HCl in dioxane (20 ml) at ambient temperature for 2 hours. The solvent was removed, and the residue was dissolved in a 1/1 mixture of ethyl acetate/hexane (30 ml) and twice concentrated. The solid was ground into powder with hexane and collected by filtration, obtaining 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (500 mg, 99%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), of 2.38 (s, 3H), 5,16 (s, 2H), 7,51 (s, 1H), 8,03 (s, 1H), 10,27 (users,3H).

Example 10-41b: tert-butyl 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylcarbamate

3,5-Dimethyl-4-((4-nitro-1H-pyrazole-1-yl)methyl)isoxazol (example 10-41c) (12 g, 53.8 mmol) and BOC anhydride (12.8 g, 64 mmol) was dissolved in 3/1/1 mixture of MeOH/EtOH/THF (300 ml) in a reaction flask Parra, and then was added 10% Pd/C (1.5 g). The mixture was shaken in hydrogenator Parra at 2 atmospheres of hydrogen for 3 hours. The mixture was filtered through a 3-inch layer of celite and concentrated on a rotary evaporator. Oil pink color was purified by chromatography on silica gel (25% ethyl acetate in hexane) to give tert-butyl 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-ylcarbamate (12,6 g, 80%) as a pink/red oil, which was hardened when standing with the formation of a light pink solid.1H NMR (CDCl3, 400 MHz): δ of 1.41 (s, 9H), 2,10 (s, 3H), 2,32 (s, 3H), of 4.90 (s, 2H), to 6.19 (users, 1H), 7,19 (s, 1H), 7,50 (s, 1H).

Example 10-41c: 3,5-dimethyl-4-((4-nitro-1H-pyrazole-1-yl)methyl)isoxazol

1H-pyrazole (10 g, 147 mmol) was added in small portions to concentrated H2SO4(100 ml)was cooled to 0ºC with bath ice/water, keeping the temperature inside the vessel below 40ºC. Caution was added dropwise to the reaction mixture concentrated HNO3(10 ml), keeping the temperature inside the vessel below 55ºC. C is the reaction mixture was heated to 55ºC and was stirred for 5 hours. The mixture was cooled to 0ºC and was carefully podlachian (pH~8) aqueous solution of NaOH (110 g of NaOH in 150 ml of H2(O) prior to the formation of a white precipitate, carefully maintaining the solution temperature below 40ºC. White solid was collected by filtration and washed with ethyl acetate/hexane (1/3), then dried under vacuum to obtain 4-nitro-1H-pyrazole (7 g, 42% yield of the selected product).13C NMR (DMSO-d6, 137,0, 126,4. To 4-nitro-1H-pyrazole (9 g, 80 mmol) in DMF (100 ml, 100 MHz) δ was added cesium carbonate (26 g, 80 mmol), and then were added 4-(chloromethyl)for 3,5-dimethylisoxazol (12.3 g, 85 mmol). The reaction mixture was stirred in DMF (100 ml) at 80ºC for 30 minutes, then cooled, diluted with H2O (150 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic phases were dried over sodium sulfate, filtered and concentrated. The residue was transferred into ethyl acetate (200 ml) and washed with H2O (2×, 100 ml). The organic phase was dried over sodium sulfate, filtered and concentrated. The solid product was ground into powder with ethyl acetate/hexane (1/9) and collected by filtration. The product was dried in high vacuum to obtain 3,5-dimethyl-4-((4-nitro-1H-pyrazole-1-yl)methyl)isoxazol (12 g, 67%) as a pale yellow solid.1H NMR (CDCl3, 400 MHz): δ of 2.23 (s, 3H), of 2.46 (s, 3H), to 5.08 (s, 2H), 8,02 (s, 1H), 8,08 (s, 1H).

Example 10-42: 5-benzyl-3-(1-((3,5-di is utilization-4-yl)methyl)-1H-pyrazole-4-yl)-1-methylimidazolidine-2,4-dione

Was obtained as in example 10-5 from 5-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-2) and iodomethane. Yield: 95%.1H NMR (CDCl3, 400 MHz): δ 2,04 (s, 3H), of 2.15 (s, 3H), 2,96 (s, 3H), 3,24 is 3.23 (m, 2 H,J=4.0 Hz), 4,23-is 4.21 (m, 1H), 5,00 (s, 2H), 7.24 to of 7.23 (m, 5H,J=4.0 Hz), of 7.70 (s, 1H), 7,87 (s, 1H).

As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.15 μm.

Example 10-43: 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylimidazolidine-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylimidazolidine-2,4-dione (example 10-41) and benzylbromide. Yield: 50%.1H NMR (CDCl3, 400 MHz): δ of 1.44 (s, 3H), 2,19 (s, 3H), 3,88 (s, 3H), 4,18 (l,J=8 Hz, 2H), 4,22 (t, 1H, J=4 Hz)), is 5.06 (s, 2H), 7,39-7,29 (m, 5H), 7,94 (s, 1H), 8,10 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.02 mm.

Example 10-44: 2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)acetic acid

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and H-Asp-OMe. Yield: 85%. MS M+H calculated 334,1; experimentally 334,1

Example 10-45: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-Piras the l-4-yl)-5-fencelineecology-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-amino-4-phenylbutanoate. Yield: 15%.1H NMR (CDCl3, 400 MHz): δ 2,09-2,02 (m, 2H), 2,19 (s, 3H), 2,41 (s, 3H), 2,83-2,78 (m, 2H), 4,13-4.09 to (t, 1H,J=8 Hz), of 5.05 (s, 2H), 5,95 (users, 1H), 7,30-7,19 (m, 5H), 7,95 (s, 1H), 8,03 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.22 μm.

Example 10-46: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(3-phenylpropyl)imidazolidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-amino-5-phenylbutanoate. Yield: 20%.1H NMR (CDCl3, 400 MHz): δ 1,72 by 1.68 (m, 1H), 1.85 to of 1.78 (m, 2H), 1,99 is 1.91 (m, 1H), 2,19 (s, 3H), 2,41 (s, 3H), 2,69 2.63 in (t,J=8 Hz, 2H), 4,13 (users, 1H), of 5.05 (s, 2H), 5,95 (users, 1H), 7,30-7,19 (m, 5H), 7,95 (s, 1H), 8,03 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.17 microns.

Example 10-47: 5-(benzoyloxymethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-1 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) and methyl-2-amino-3-(benzyloxy)propanoate. Yield: 32%.1H NMR (CDCl3, 400 MHz): δ 2,19 (s,3H), 2,43 (s, 3H), 3,70-to 3.67 (m,1 H), 3,89-3,86 (m, 1H), or 4.31-4,30 (m, 1H), 4,56-4,32 (l,J=1,6 Hz, 2H), of 5.05 (s, 2H), 5,62 (users, 1H), 7,35-7,29 (m, 5H), 7,88 (s, 1H), 8,04 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,76 ám.

Example 10-48: 2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)-N-phenylacetamide

3-(1-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)acetic acid (example 10-44) (100 mg, 0.3 mmol), aniline (33 mg, 0.36 mmol), PyBOP (hexaphosphate benzotriazol-1-yl-hydroxy-respirology-phosphonium) (187 mg, 0.36 mmol) and triethylamine (0.05 ml, 0.36 mmol) were mixed in DMF (1 ml). The reaction mixture was stirred at 65ºC for 4 hours. The reaction mixture was left to cool to room temperature and then was diluted with ethyl acetate (2 ml). The organic phase was washed with saturated sodium bicarbonate solution (2×, 2 ml) and then saturated NaCl solution (1 ml). The organic phase was extracted, dried over anhydrous Na2SO4and filtered. The crude product is re-suspended in MeOH (1 ml) and purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O; 16-minute gradient). Pure fractions were combined and solvent was removed on a rotary evaporator to obtain 2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-deok imidazolidin-4-yl)-N-phenylacetamide in the form of a white solid (50%). 1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H), 2.40 a (s, 3H), of 2.72 (m, 1H), 3,14-3,13 (d, 1H,J=4 Hz), 4,54-4,51 (l,J=8 Hz, 1H), 5,04 (s, 2H), 6,53 (users, 1H), 7,15-7,13 (m, 1H), 7,33-7,22 (m, 2H), 7,47 was 7.45 (d,J=8 Hz, 2H), 7,78 (users, 1H), to 7.09 (s, 1H), with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.75 ám.

Example 10-49: N-benzyl-2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)ndimethylacetamide

Was obtained as in example 10-48 3-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)acetic acid (example 10-44) and benzylamine. Yield: 30%.1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H), 2,41 (s, 3H), 2,56-2,52 (m, 1H,J=16 Hz), 2,56-2,52 (m, 1H), 3.00 and-2,96 (m, 1H), 4,45-of 4.44 (d,J=5.6 Hz, 2H), 5,04 (s, 2H), 5,96 (users, 1H), 6,36 (users, 1H), was 7.36-7,25 (m, 5H), of 7.90 (s, 1H, with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 1.3 μm.

Example 10-50: 2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)-N-(3-methoxybenzyl)ndimethylacetamide

Was obtained as in example 10-48 3-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)acetic acid (example 10-44) and (3-methoxyphenyl)methanamine. Yield: 50%. LC/MS; the expected value 453; experimentally 453,1. As shown, the above-mentioned soy is inania inhibits bitter taste receptor hT2R08 and has the IC 501,7 mm.

Example 10-51: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methylimidazolidine-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) (50 mg, of 0.182 mmol) and cesium carbonate (60 mg, 0.185 mmol) were mixed in DMF (1 ml) for 15 minutes in a nitrogen atmosphere at room temperature. Then added logmean (14 mg, 0.185 mmol) and continued stirring the reaction mixture for an additional 2 hours. Added H2O (2 ml) and product was extracted with ethyl acetate (1 ml, 2×). The organic phase was collected and washed with a saturated solution of sodium bicarbonate(2 ml, 2×), dried and filtered. The solvent was removed in a stream of nitrogen and then further dried under high vacuum to obtain 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methylimidazolidine-2,4-dione as a white solid (42 mg, 80%). Yield: 80%.1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H), 2,41 (s, 3H), 3,06 (s, 3H), of 3.95 (s, 2H), of 5.05 (s, 2H), 7,89 (s, 1H), with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.58 μm.

Example 10-52: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,5,5-trimethylindolenine-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylimidazolidine-2,4-dio is (example 10-41) (50 mg, 0,173 mmol) and 60% NaH (8 mg, 0,190 mmol) were mixed in DMF (1 ml) for 30 minutes. Added MeI (0.04 ml, 0,190 mmol) and the reaction mixture was stirred an additional 4 hours. The reaction mixture was acidified using 1N HCl and was diluted with ethyl acetate (2 ml). The organic phase was dried, filtered and solvent was removed in a stream of nitrogen. The crude product is re-suspended in MeOH (1 ml) and purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: a 16-minute gradient). Pure fractions were combined and solvent was removed under vacuum to obtain 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,5,5-trimethylindolenine-2,4-dione as a white solid (25 mg, 50%).1H NMR (CDCl3, (CDCl3, 400 MHz): δ 1,45 (s, 6H), are 2.19 (s, 3H), 2,42 (s, 3H), equal to 2.94 (s, 3H), of 5.05 (s, 2H), 7,92 (s, 1H), 8,08 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,80 µm.

Example 10-53: 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and benzylbromide. Yield: 40%.1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H), 2,42 (s, 3H), of 3.84 (s, 2H), br4.61 (s, 2H), is 5.06 (s, 2H), 7,40-7,27 (m, 5H), 7,92 (s, 1H), 8,08 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT208 and has the IC 500,09 ám.

Example 10-54: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(pyridine-2-ylmethyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione and 2-(methyl bromide)pyridine. Yield: 50%.1H NMR (CDCl3, 400 MHz): δ 2,19 (s, 3H), 2,41 (s, 3H), of 4.12 (s, 2H), 4,71 (s, 2H), of 5.05 (s, 2H), 7,72-of 7.23 (m, 4H), 7,92 (s, 1H), 8,08 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.68 μm.

Example 10-55: 1-((3,5-dimethylisoxazol-4-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione and 4-(chloromethyl)for 3,5-dimethylisoxazole. Yield: 50%.1H NMR (CDCl3, 400 MHz): δ 2,19 (s, 3H), and 2.27 (s, 3H), 2,43-2,42 (l,J=5,2 Hz, 6H), 3,82 (s, 2H), and 4.40 (s, 2H), of 5.05 (s, 2H), 7,92 (s, 1H), with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,04 ám.

Example 10-56: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(3-methoxybenzyl)imidazolidin-2,4-dione

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (400 mg, of 2.08 mmol), piperidin-2-yl carbonate (450 mg, of 2.08 mmol) and triethylamine (0,290 ml of 2.08 mmol who) was stirred in dichloromethane (7 ml) for 12 hours at room temperature. The reaction mixture was concentrated under vacuum to obtain 4-((4-isocyanato-1H-pyrazole-1-yl)methyl) - for 3,5-dimethylisoxazole as off-white solid in quantitative yield. Was added ethanol (1 ml) with methyl 2-amino-3-(3-methoxyphenyl)propanoate (68 mg, 0,327 mmol) and triethylamine (0,064 ml, 0,461 mmol). The reaction mixture was stirred while boiling under reflux for 12 hours, then left to cool to room temperature. The solvent was removed in a stream of nitrogen. The crude product is re-suspended in MeOH (1 ml) and purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: a 16-minute gradient). Pure fractions were combined and solvent was removed under vacuum to obtain 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(3-methoxybenzyl)imidazolidin-2,4-dione as a white solid, yield: 50%.1H NMR (CDCl3, 400 MHz): δ 2,19 (s, 3H), 2,41 (s, 3H), 3,34-to 3.33 (d,J=3.2 Hz, 1H), 3,31-3,29 (l,J=8 Hz, 1H), 3,76 (s, 3H), 4,33-4,30 (m, 1H), of 5.05 (s, 2H), 5,95 (users, 1H), 7,25-7,21 (t, 1H), 6,82-of 6.78 (m, 3H), a 7.85 (s, 1H), to 7.99 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.13.

Example 10-57: 5-(cyclohexylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-56 from 1-((3,5-demetrise Sasol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 10-41a) and methyl-2-amino-3-cyclohexylpropionate. Yield: 30%.1H NMR (CDCl3, 400 MHz): δ 1,06-of 0.95 (m, 2H), 1,29-of 1.15 (m,3H), 1.60-to 1,50 (1H) 1,77-1,67 (7 H), 1,91-of 1.85 (m, 1H), 2,19 (s, 3H), 2,41 (s, 3H), 4,19-to 4.15 (m, 1H), of 5.05 (s, 2H), 6,01 (users, 1H), to $ 7.91 (s, 1H), with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.96 μm.

Example 10-58: 5-(cyclopentylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-56 from 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 10-41a) and methyl-2-amino-3-cyclopentylpropionate. Yield: 50%.1H NMR (CDCl3, 400 MHz): δ 1,20-to 1.14 (m, 3H), 1,68-of 1.55 (m, 6H), 2,04-of 1.92 (m, 2H), 2,19 (s, 3H), 2,42 (s, 3H), 4,14-4,11 (m, 1H), of 5.05 (s, 2H), 5,52 (users, 1H), of 7.90 (s, 1H), of 8.06 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,31 mm.

Example 10-59: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(4-hydroxybenzyl)imidazolidin-2,4-dione

Was obtained as in example 10-56 from 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 10-41a) and methyl-2-amino-3-(4-hydroxyphenyl)propanoate. Yield: 50%.1H NMR (CDCl3, 400 MHz): δ is 2.41 (s, 3H), 2,85 (s, 3H), 3,26-3,25 (d, 1H, J=4 Hz), 3,23-3,22 (l,J=4 Hz, 1H) or 4.31-to 4.28 (m, 1H), 5,04 (s, 2H), 5,77-5,74 (users, 1H), 7,07? 7.04 baby mortality (l,J=12 Hz, 2H), 's 6.75 6.73 x (l,J=8 Hz, 2H), 7,06 (s, 1H), of 7.97 (s, 1H), 9,43 (users, 1H). As pok is connected, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.33 μm.

Example 10-60: 5-(3,4-dihydroxybenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-56 from 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 10-41a) and methyl-2-amino-3-(3,4-dihydroxyphenyl)propanoate. Yield: 50%. MS M+H calculated 398,1; experimentally 398,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.51 μm.

Example 10-61: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-isobutylthiazole-2,4-dione

Was obtained as in example 10-41 from 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 10-41a) and ethyl-2-isocyanato-4-methylpentanoate. Yield: 50%.1H NMR (CDCl3, 400 MHz): δ 1,01-0,98 (m, 8H), 1,87-to 1.82 (m, 1H), 2,19 (s, 3H), 2,41 (s, 3H), 4,13-4,12 (t, 1H), (of 5.05 (s, 2H), 5,70 (users, 1H), of 7.90 (s, 1H), with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 1.0 μm.

Example 10-62: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-isopropylimidazole-2,4-dione

Was obtained as in example 10-41 from 1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine hydrochloride (example 10-41a) and e is the Il-2-isocyanato-3-methylbutanoate. Yield: 30%.1H NMR (CDCl3, 400 MHz): δ 0,96-of 0.94 (d, 3H, J=7.2 Hz), 1,09-of 1.07 (d, 3H, J=8 Hz), 2,19 (s, 3H), 2.26 and-2,22 (m, 1H), 2.40 a (s, 3H), was 4.02 (s, 1H), of 5.05 (s, 2H), of 5.53 (users, 1H), of 7.90 (s, 1H), with 8.05 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501.1 µm.

Example 10-63: (Z)-5-benzylidene-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) (275 mg, 1 mmol), benzaldehyde (140 mg, 1.3 mmol) and sodium acetate (205 mg, 2.5 mmol) in glacial acetic acid (3 ml) was exposed to radiation in a microwave reactor for 7 hours at 185°C. After cooling, the mixture was diluted with H2O (100 ml) and was extracted with ethyl acetate (3×50 ml). The combined organic extracts were washed with saturated aqueous solution of sodium carbonate, dried over sodium sulfate, filtered and concentrated. The solid product was ground into powder with ethyl acetate/hexane (1/1) and dried under high vacuum to obtain (Z)-5-benzylidene-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (173 mg, 48%) as a pale yellow solid.1H NMR (DMSO-d6, 400 MHz): δ 2,14 (s, 3H), 2.40 a (s, 3H), 6,59 (s, 1H), 5,20 (s, 2H), 7,33-7,40 (m, 3H), 7,66 (s, 2H), 7,81 (s, 1H), 8,21 (s, 1H), br11.01 (users, 1H). As shown, the above-mentioned connection is giving inhibits bitter taste receptor hT2R08 and has the IC 500.34 micron.

Example 10-64: 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-1,2,4-triazolin-3,5-dione

Ethyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-3,5-dioxo-1,2,4-triazolin-1-carboxylate (3.2 g, 8,8 mmol) was stirred in (1/1) mixture of MeOH/1N aqueous NaOH (100 ml) at ambient temperature for 30 minutes. The mixture was acidified using 1N aqueous HCl (150 ml), extracted with ethyl acetate (3×, 100 ml), dried over sodium sulfate, filtered and concentrated to obtain 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-1,2,4-triazolin-3,5-dione (2.3 g, 89%) as a yellow solid.1H NMR (DMSO-d6, 400 MHz): δ to 2.18 (s, 3H), of 2.44 (s, 3H), 3,05 (s, 3H), of 5.17 (s, 2H), 7,94 (s, 1H), 8,13 (s, 1H).

Example 10-64a: Ethyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-3,5-dioxo-1,2,4-triazolin-1-carboxylate

Ethylchloride (1.3 g, 12 mmol) was added to a mixture of N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methylhydroperoxide (example 10-64b) (2.5 g, 9 mmol) and triethylamine (1.2 g, 12 mmol) in acetonitrile (100 ml). The mixture was boiled under reflux for 1 hour, cooled, then diluted with 1N aqueous HCl (150 ml) and was extracted with ethyl acetate (3×, 75 ml). The combined organic extracts were dried over sodium sulfate, filtered and concentrated is on a rotary evaporator. The solid was ground into powder with ethyl acetate/hexane (1/3), and dried under high vacuum to obtain ethyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-3,5-dioxo-1,2,4-triazolin-1-carboxylate (3.2 g, 94%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 1.28 (in t,J=7.2 Hz, 3H), 2.13 and (s, 3H), 2.40 a (s, 3H), 3,24 (s, 3H), 4,30 (t,J=7.2 Hz, 2H), total of 5.21 (s, 2H), 7,73 (m, 1H), 8,16 (s, 1H).

Example 10-64b: N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methylhydrosiloxane

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) (1.25 g, 5.3 mmol) was stirred in toluene (30 ml) at the boiling temperature under reflux for 40 minutes. The mixture was cooled to ambient temperature and was added methylhydrazine (0.3 ml, 260 mg, 5.6 mmol) and the mixture is boiled under reflux for 30 minutes. After cooling the reaction to room temperature, the solvent was removed on a rotary evaporator and the solid product was ground into powder with ethyl acetate/hexane (2/5), and dried under high vacuum to obtain N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methylhydroperoxide (1.1 g, 79%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ is 2.09 (s, 3H), of 2.36 (s, 3H), 2,98 (s, 3H), br4.61 (s, 2H), 5,02 (s, 2H), 7,42 (s, 1H), 7,72 (m, 1H), 8,78 (s, 1H).

Example 10-65: 1-Benzyl-4-(1-((3,5-dime isoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-1,2,4-triazolin-3,5-dione

4-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-1,2,4-triazolin-3,5-dione (example 10-64) (785 mg, 2.7 mmol) was dissolved in acetonitrile (50 ml). Was added triethylamine (1 g, 10 mmol) and benzylbromide (510 mg, 3 mmol) and the reaction mixture was stirred at ambient temperature for 12 hours. Then the mixture was concentrated on a rotary evaporator, dissolved in methanol (5 ml) and was purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: 25 minute gradient). Pure fractions were combined and concentrated and the product recrystallized from ethanol to obtain 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-1,2,4-triazolin-3,5-dione (210 mg, 20%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2.13 in (s, 3H), 2,39 (s, 3H), to 3.09 (s, 3H), to 4.81 (s, 2H), by 5.18 (s, 2H), 7,30-to 7.35 (m, 5H), 7,76 (s, 1H), 8,18 (s, 1H). MS M+H calculated 381,1; experimentally 381,1. Melting point: 124-126ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.02 mm.

Example 10-66: 1-Benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-1,2,4-triazolin-3,5-dione

Was obtained as in example 10-65 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-1,2,4-triazolin-3,5-dione (example 10-64) and 1-(2-bromacil)-4-fervently. Output is: 14%. 1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H), 2.40 a (s, 3H), 2,87 (t,J=6,8 Hz, 2H), 3,14 (s, 3H), 3,83 (t,J=7.2 Hz, 2H), to 5.03 (s, 2H), 6,95 (t,J=8,4 Hz, 2H), 7,14 (t,J=8 Hz, 2H), to 7.77 (s, 1H), 7,95 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.01 µm.

Example 10-67: 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-2-(2-phenoxyethyl)-1,2,4-triazoline-3,5-dione

Was obtained as in example 10-65 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-1,2,4-triazolin-3,5-dione (example 10-64) and (2 bromoethoxy)benzene. Yield: 20%.1H NMR (DMSO-d6, 400 MHz): δ 2.13 in (s, 3H), 2.40 a (s, 3H), 3.15 in (s, 3H), 3,99 (t,J=4.4 Hz, 2H), 4,13 (t,J=4,8 Hz, 2H), 5,20 (s, 2H), 6,80 (l,J=8 Hz, 2H), 6.90 to (t,J=7,1 Hz, 1H), 7,22 (t,J=8 Hz, 2H), of 7.75 (s, 1H), 8,17 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,031 mm.

Example 10-68: 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-carbonylated (example 10-1a) (1 g, 4.1 mmol) was boiled under reflux in toluene (100 ml) for one hour. The reaction mixture was cooled to room temperature and was added acylhydrazines (0.45 g, 43 mmol). The reaction mixture was heated to boiling under reflux is stirred for 1 hour, then cooled and concentrated on a rotary evaporator. The residue was transferred into ethanol (100 ml) was added potassium carbonate (100 mg). The mixture was boiled under reflux for 12 hours, then filtered, cooled to ambient temperature and neutralized with acetic acid (about 7 drops). The solvent was removed on a rotary evaporator and the resulting solid was ground into powder with ethyl acetate/hexane (1/9) to obtain 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (0,98 g, 85%) as off-white solid.1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H), 2,41 (s, 3H), to 4.98 (s, 2H) 7,16 (s, 1H), 7,38 (s, 1H).

Example 10-69: 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2-dimethyl-1,2,4-triazolin-3,5-dione

4-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (example 10-68) (100 mg, 0.36 mmol), methyliodide (141 mg, 1 mmol) and cesium carbonate (325 mg, 1 mmol) was stirred in a 2/1 mixture of acetonitrile/DMF (5 ml) at ambient temperature for 2 hours. The mixture was diluted with aqueous 1N HCl (100 ml) and was extracted with ethyl acetate (3×50 ml). The combined organic extracts were dried over sodium sulfate, concentrated, and the crude residue was transferred into MeOH and purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: a 25-minute hail the UNT). Pure fractions were combined and concentrated to obtain 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2-dimethyl-1,2,4-triazolin-3,5-dione (89 mg, 80%) as a transparent semi-solid substances.1H NMR (CDCl3, 400 MHz): δ to 2.18 (s, 3H), 2,41 (s, 3H), up 3.22 (s, 6H), 5,04 (s, 2H), 7,88 (s, 1H), 8,03 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.6 microns.

Example 10-70: 1,2-dibenzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione

Was obtained as in example 10-65 4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2-dimethyl-1,2,4-triazolin-3,5-dione (example 10-69) and benzylbromide. Yield: 69%.1H NMR (CDCl3, 400 MHz): δ 2,14 (s, 3H), of 2.36 (s, 3H)and 4.65 (s, 4H), at 4.99 (s, 2H), 7,06-was 7.08 (m, 4H), 7,19-7,25 (m, 6H), 7,86 (s, 1H), 8,02 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.8 μm.

Example 10-71: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((6-(hydroxymethyl)pyridine-2-yl)methyl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione and 6-methyl bromide-2-pyridinemethanol. Yield: 35%. MS M+H calculated 397,2; experimentally 397,2. As shown, the above-mentioned compound inhibits the receptor Gorkog the taste hT2R08 and has the IC 50to 0.72 microns.

Example 10-72: 1-((3,4-dimethoxypyridine-2-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-5 from 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione and 3,4-dimethoxy-2-chloromethylpyridine hydrochloride. Yield: 26%. MS M+H calculated 360,2; experimentally 360,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 1.0 μm.

Example 10-73: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((6-(tetrahydrofuran-2-yl)methyl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione and tetrahydrofurfurylamine. Yield: 28%. MS M+H calculated 427,2; experimentally 427,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501.4 µm.

Example 10-74: 1-(cyclohexylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and bromeilles. Yield: 20%.1H NMR (DMSO-d6, 400 MHz): δ 0,88 (kV,J=10.4 Hz, 2H), 1,09-1,19 (who, 3H), 1,58-of 1.65 (m, 6H), 2,12 (s, 3H), of 2.38 (s, 3H), 3,13 (l,J=7.2 Hz, 2H), 4,06 (s, 2H), 5,17 (s, 2H), of 7.75 (s, 1H), 8,14 (s, 1H). MS M+H calculated 372,2; experimentally 372,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.28 μm.

Example 10-75: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((6-methoxypyridine-2-yl)methyl)imidazolidin-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (100 mg, 0.4 mmol), (6-methoxypyridine-2-yl)methanol (example 10-75a) (101 mg, 0.7 mmol), tributylphosphine (147 mg, 0.7 mmol) and 1,1""-azobis(N,N-dimethylformamide) (125 mg, 0.7 mmol) was dissolved in THF (5 ml) and stirred at room temperature for 15 hours. The reaction mixture was diluted with brine (100 ml) and was extracted with ethyl acetate (2×, 100 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated on a rotary evaporator. The residue was transferred to a methanol (5 ml) and was purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: 25 minute gradient). Pure fractions were combined, concentrated, and then re-dissolved in ethyl acetate/hexane (1:9). The solution was cooled at 5ºC for 15 hours, while the formed white solid. The precipitate was collected to obtain 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)1-((6-methoxypyridine-2-yl)methyl)imidazolidin-2,4-dione (5 mg, 4%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), is 2.37 (s, 3H), 3,74 (s, 3H), 4,17 (s, 2H), of 4.54 (s, 2H), 5,17 (s, 2H), 6,69 (l,J=7,6 Hz, 1H), of 6.96 (d,J=6,8 Hz, 1H), to 7.67 (d,J=6,8 Hz, 1H), 7,76 (s, 1H), 8,17 (s, 1H). MS M+H calculated 397,2; experimentally 397,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50to 0.23 μm.

Example 10-75a: (6-methoxypyridine-2-yl)methanol

Methyl-6-methoxypyridine-2-carboxylate (2 g, 11,96 mmol) in anhydrous methanol (20 ml) was cooled to 0ºC in nitrogen atmosphere and the solution was slowly added sodium borohydride (1,36 g, 35,89 mmol). The reaction mixture was left under stirring at 0ºC for 30 minutes, then left to warm to room temperature for 1 hour. The reaction mixture was extinguished with water and concentrated on a rotary evaporator. The reaction mixture was diluted with brine (100 ml) and was extracted with a solution of dichloromethane/2-propanol(2:1) (3×, 150 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated on a rotary evaporator to obtain (6-methoxypyridine-2-yl)methanol (500 mg, 30%) as oil. MS M+H calculated 140,1; experimentally 140,1.

Example 10-76: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-methoxyphenethyl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 6) and 2-methoxyphenethylamine. Yield: 52%.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), of 2.38 (s, 3H), 2,80 (t,J=7.2 Hz, 2H), 3,51 (t,J=7.2 Hz, 2H, in), 3.75 (s, 3H), a 4.03 (s, 2H), 5,17 (s, 2H), 6,85 (t,J=7.2 Hz, 1H), 6,95 (l,J=8,4 Hz, 1H), 7,16-7,21 (m, 2H), 7,73 (s, 1H), 8,13 (s, 1H). MS M+H calculated 410,2; experimentally 410,1. Melting point: 97-98ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,14 m.

Example 10-77: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-florfenicol)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3-fortunaticlomid. Yield: 22%.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), of 2.38 (s, 3H), 2,80 (t,J=7.2 Hz, 2H), 3,57 (t,J=7.2 Hz, 2H), 4,06 (s, 2H), 5,17 (s, 2H), 6,85 (dt,J=to 8.4, 2.0 Hz, 1H), 7,11 (t,J=8,4 Hz, 2H), 7,32 (kV,J=7.8 Hz, 1H), 7,73 (s, 1H), 8,12 (s, 1H). MS M+H calculated 398,2; experimentally 398,1. Melting point: 110-111ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-78: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-fencelineecology-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and geneticbased. Yield: 37%.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), of 2.38 (s, 3H), and 2.83 (t,J=6,4 Hz, 2H), 3,55 (t,J=7,4 Hz, 2H), a 4.03 (s, 2H), 5,17 (s, 2H), 7.18 in-7,30 (m, 5H), 7,73 (s, 1H), 8,13 (s, 1H). MS M+H calculated 380,2; experimentally 380,1. Melting point: 95-96ºC. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,14 m.

Example 10-79: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-methoxyphenethyl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 4-methoxyphenethylamine. Yield: 32%.1H NMR (DMSO-d6, 400 MHz): δ 2,11 (s, 3H), of 2.38 (s, 3H), 2,78 (t,J=7,4 Hz, 2H), 3,50 (t,J=7,4 Hz, 2H), 3,69 (s, 3H), was 4.02 (s, 2H), 5,16 (s, 2H), 6,84 (l,J=8,4 Hz, 2H), 7,15 (l,J=8,4 Hz, 2H), 7,73 (s, 1H), 8,12 (s, 1H). MS M+H calculated 410,18; experimentally 410,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,04 ám.

Example 10-80: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-naphthalene-1-yl)ethyl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H/i> -pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 1-(2-bromacil)naphthalene. Yield: 20%. MS M+H calculated 430,18; experimentally 430,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC501,27 µm.

Example 10-81: 1-(2-chlorphenyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 2-chlorophenethylamine. Yield: 25%. MS M+H calculated 414,13; experimentally level of 414.2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.25 μm.

Example 10-82: 1-(3-chlorphenyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3-chlorophenethylamine. Yield: 27%. MS M+H calculated 414,13; experimentally level of 414.2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,20 ám.

Example 10-83: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-florfenicol)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethy is isoxazol-4-yl)methyl)-1 H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 2-fortunaticlomid. Yield: 24%. MS M+H calculated 398,16; experimentally 398,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.13.

Example 10-84: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-florfenicol)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 4-fortunaticlomid. Yield: 34%. MS M+H calculated 398,16; experimentally 398,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.01 µm.

Example 10-85: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-methoxyphenethyl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3-methoxyphenethylamine. Yield: 34%. MS M+H calculated 410,18; experimentally 410,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.16 μm.

Example 10-86: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-hydroxyphenethyl)imidazolidin-2,4-dione

Was obtained as in example 10-52 -(1-((3,5-dimethylisoxazol-4-yl)methyl)-1 H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 4-hydroxyphenylethylamine. Yield: 31%. MS M+H calculated 396,16; experimentally 396,1. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,41 mm.

Example 10-87: 1-(3,4-dimethoxyphenethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and 3,4-dimethoxyphenethylamine. Yield: 36%. MS M+H calculated 440,19; experimentally 440,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.26 μm.

Example 10-88: 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2-he

1-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2-he (example 10-88a) (50 mg, 0,19 mmol) and 60% sodium hydride (8 mg, 0.21 mmol) in DMF (3 ml) was stirred at room temperature for 15 minutes, then was cooled to 0ºC. To the mixture was added benzylbromide (33 mg, 0,19 mmol) and left to warm at room temperature for 2 hours. The reaction was suppressed with methanol and then concentrated on a rotary evaporator. The reaction mixture was diluted with brine (50 ml) and was extracted with dichlor the tan (2×, 50 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) to give 1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2-it (21 mg, 31%) as a white solid. MS M+H calculated 352,17; experimentally 352,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.71 μm.

Example 10-88a: 1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2-he

1-(2-Chloroethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)1H-pyrazole-4-yl)urea (example 10-88b) (115 mg, 0,39 mmol) and 60% sodium hydride (17 mg, 0.42 mmol) in DMF (2 ml) was stirred at 0ºC for 15 minutes, then left to warm to room temperature while stirring for 2 hours. The reaction was suppressed with methanol and then concentrated on a rotary evaporator. The reaction mixture was diluted with brine (50 ml) and was extracted with dichloromethane (2×50 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30-indutny gradient) to give 1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1 H-pyrazole-4-yl)imidazolidin-2-she (98 mg, 97%) as a white solid.1H NMR (DMSO-d6, 400 MHz): δ 2,10 (s, 3H), is 2.37 (s, 3H), 3,35-3,39 (m, 2H), 3,61 (l,J=8,8 Hz, 1H), 3,63 (l,J=10.4 Hz, 1H), 5,07 (s, 2H), of 6.71 (s, 1H), 7,42 (s, 1H), 7,74 (s, 1H). MS M+H calculated 262,12; experimentally 262,1.

Example 10-88b: 1-(2-chloroethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)urea

1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-amine (419 mg, to 2.18 mmol) and 2-chlorotriazine (230 mg, to 2.18 mmol) in acetonitrile (5 ml) was heated at 65ºC for 16 hours. The reaction mixture was cooled to room temperature and concentrated on a rotary evaporator. The residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient), was dried and ground into powder with ethyl acetate/hexane (1/9) to give (1-(2-chloroethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)1H-pyrazole-4-yl)urea (258 mg, 40%) as a yellow solid. MS M+H calculated 298,10; experimentally 298,1

Example 10-89: 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(methoxymethyl)-1,2,4-triazoline-3,5-dione

Was obtained as in example 10-91 from 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (example 10-91a) and bromatologia ether. Output: 8%. MS M+H calculated 411,17; experimentally 411,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.02 mm.

Example 10-90: 1-((1,3-dimethyl-1H-pyrazole-4-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (10a) (200 mg, 0.7 mmol), 4-(chloromethyl)-1,3-dimethyl-1H-pyrazole (144 mg, 1 mmol) and cesium carbonate (325 mg, 1 mmol) was dissolved in 2 ml of DMF and subjected to the action of radiation in a microwave reactor at 165ºC for 5 minutes. The reaction mixture was cooled to room temperature, and removing the precipitate in the form of salts by filtration. Received a clear solution containing the crude product was purified by HPLC (10-95% acetonitrile/water; 25 minutes) to give 1-((1,3-dimethyl-1H-pyrazole-5-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (150 mg, 53%) as a light brown solid.1H NMR (DMSO, 400 MHz): δ is 2.09 (s, 3H), and 2.14 (s, 3H), of 2.20 (s, 3H), 3,70 (s, 3H), 3,99 (s, 2H), 4,55 (s, 2H), 5,19 (s, 2H), 6,06 (s, H), to 7.77 (s, H), 8,17 (s, H). MS M+H calculated 384,2; experimentally 384,2. Melting point: 145-146ºC.

Example 10-91: 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-ethyl-1,2,4-triazolin-3,5-dione

1-Benzyl-4-(1-((5-dimethylisoxazol-4-yl)methyl)-1 H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (100 mg, 0.27 mmol), bromate (149 mg, of 1.36 mmol) and cesium carbonate (355 mg, 1.1 mmol) in DMF (5 ml) was heated at 80ºC for 15 hours. The reaction mixture was cooled to room temperature, diluted with brine (50 ml) and was extracted with ethyl acetate (2×50 ml). The combined organic extracts were dried over magnesium sulfate, filtered and concentrated on a rotary evaporator. The residue was purified by HPLC (5-95% acetonitrile in H2O: 25 minute gradient) to give 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-ethyl-1,2,4-triazolin-3,5-dione (40 mg, 37%) as oil.1H NMR (DMSO-d6, 400 MHz): δ 0,97 (t,J=7.2 Hz, 3H), 2.13 and (s, 3H), 2,39 (s, 3H), to 3.58 (kV,J=6,8 Hz, 2H), 4,80 (s, 2H), by 5.18 (s, 2H), 7,28 and 7.36 (m, 5H), to 7.77 (s, 1H), 8,19 (s, 1H). MS M+H calculated 395,18; experimentally 395,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,04 ám.

Example 10-91a: 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione

1-Benzyl-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)hydrazinecarboxamide (2.00 g, 5,88 mmol), ethylchloride (6,38 g, 58,77 mmol) and triethylamine (1.78 g, 17,63 mmol) in acetonitrile (50 ml) was heated at 100ºC for 48 hours. The mixture was cooled to 80º C, was added 1 M NaOH (aqueous) (5 ml) is stirred the reaction mixture for 1 hour. The reaction mixture was cooled to room temperature and concentrated on a rotary evaporator. The residue was dissolved in dichloromethane and filtered to remove salts, and the solution was concentrated to obtain 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (1.28 g, 60%) as a yellow oil. MS M+H calculated 367,14; experimentally 367,2.

Example 10-91b: 1-benzyl-N-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)hydrazinecarboxamide

1-((3,5-Dimethylisoxazol-4-yl)methyl-1H-pyrazole-4-carbonylated (2,79 g, 11,33 mmol) in toluene (70 ml) was heated at the boil under reflux for 4 hours to obtain 4-((4-isocyanato-1H-pyrazole-1-yl)methyl) - for 3,5-dimethylisoxazolein situ. To the mixture was added benzoylhydrazone the dihydrochloride (2,44 g, 12,45 mmol) and triethylamine (to 2.29 g, 22,64 mmol). The mixture was heated at 100ºC for 4 more hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (150 ml) and filtered through celite. Then the mother liquor was washed brine (150 ml) and the organic phase was dried over magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel (100%-90% dichloromethane in methanol: 30 minute gradient) to give 1-benzyl-N-(1-((3,5-dimethylisoxazol-4-yl)IU the Il)-1 H-pyrazole-4-yl)hydrazinecarboxamide (2.00 g, 52%) as oil. MS M+H calculated 341,16; experimentally 341,2.

Example 10-92: 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(2-methoxyethyl)-1,2,4-triazoline-3,5-dione

Was obtained as in example 10-91 from 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (example 10-91a) and 2-pomatoleios ether. Yield: 20%. MS M+H calculated 425,19; experimentally 425,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-93: 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-propyl-1,2,4-triazolin-3,5-dione

Was obtained as in example 10-91 from 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (example 10-91a) and 1-bromopropane. Yield: 38%. MS M+H calculated 409,19; experimentally 409,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500.06 micron.

Example 10-94: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-phenylpropyl)imidazolidin-2,4-dione

Was obtained as in example 10-52 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) and (3-bromopropyl)benzene. In the progress: 36%. MS M+H calculated 394,2; experimentally 394,2. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC50of 0.26 μm.

Example 10-95: 1-benzyl-2-butyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl-1,2,4-triazolin-3,5-dione

Was obtained as in example 10-91 from 1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione (example 10-91a) and 1-bromobutane. Yield: 22%. MS M+H calculated 423,21; experimentally 423,15. As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,41 mm.

Example 10-96: tert-butyl 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzylcarbamoyl

3-(1-((3,5-Dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione (example 10-1) (0,70 mg, 0,254 mmol), tert-butyl 3-(hydroxymethyl)benzylcarbamoyl (0,254 mmol, 60 mg), diethylazodicarboxylate (0.50 mmol, 86 mg) and P-tBu3(125 ml, 0.50 mmol) was stirred in THF (1 ml) for 4 hours. The reaction mixture was diluted with ethyl acetate (1.5 ml) and washed with saturated sodium bicarbonate solution (2×, 1.5 ml). Collected organic phase and concentrating the mixture in a stream of nitrogen. The crude product was purified by the method of column chromatography on silica gel, using etelaat is as eluent. Pure fractions were combined and solvents removed on a rotary evaporator to obtain tert-butyl 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzylcarbamoyl in the form of a white solid (112 mg, 90%).1H NMR (CDCl3, 400 MHz): δ of 1.44 (s, 9H), are 2.19 (s, 3H), 2,42 (s, 3H), 3,48 (users, 1H), 3,84 (s, 2H), or 4.31-4,30 (l,J=6 Hz, 1H), 4,60 (s, 2H), 4,87 (users, 1H), is 5.06 (s, 2H), was 7.36-7,16 (m, 4H), 7,92 (s, 1H), 8,08 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC500,90 µm.

Example 10-97: 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(hydroxymethyl)imidazolidin-2,4-dione

4-((4-Isocyanate-1H-pyrazole-1-yl)methyl) - for 3,5-dimethylisoxazol (example 10-1) (784 mg, 3.6 mmol), hydrochloride of the methyl ester of serine (672 mg, 4,32 mmol) and triethylamine (1 ml, 7.2 mmol) was boiled under reflux in toluene (16 ml) for 8 hours. The reaction mixture was left to cool to room temperature and then the solution was concentrated on a rotary evaporator. The product was purified by the method of reversed-phase HPLC (5-95% acetonitrile in H2O: a 16-minute gradient) to give 3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(hydroxymethyl)imidazolidin-2,4-dione as a white solid (60 mg, 25%).1H NMR (CDCl3, 400 MHz): δ (s, 3H), 2.40 a (s, 3H), 3,13-of 3.07 (m, 1H), 3,94-3,93 (l,J=4 G is, 2H), 4,21-4,19 (t,J=4 Hz, 1H), to 5.03 (s, 2H), 6,68 (users, 1H), 7,87 (s, 1H), to 7.99 (s, 1H). As shown, the above-mentioned compound inhibits bitter taste receptor hT2R08 and has the IC503 microns.

No. of connectionsConnectionhT2R8 IC50(µm)
10-66
4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-florfenicol)-2-methyl-1,2,4-triazolin-3,5-dione
0,012
10-15
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(methylthio)benzyl)imidazolidin-2,4-dione
0,016
10-42
5-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methylimidazolidine-2,4-dione
0,017
10-26
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-methylbenzyl)imidazolidin-2,4-dione
0,019

10-43
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylimidazolidine-2,4-dione
0,020
10-89
1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(methoxymethyl)-1,2,4-triazoline-3,5-dione
0,024
10-10
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-hydroxybenzyl)imidazolidin-2,4-dione
0,026
10-84
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-florfenicol)imidazolidin-2,4-dione
0,027
10-91
1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-ethyl-1,2,4-triazolin-3,5-dione
0,041

10-12
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-hydroxybenzyl)imidazolidin-2,4-dione
0,043
10-79
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-methoxyphenethyl)imidazolidin-2,4-dione
0,044
10-31
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-terbisil)imidazolidin-2,4-dione
0,045
10-32
4-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzonitrile
0,048
10-35
1-(3,4-dimethoxybenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,051

10-36
1-(benzo[d][1,3]dioxol-5-ylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,053
10-92
1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-(2-methoxyethyl)-1,2,4-triazoline-3,5-dione
0,057
10-27
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-terbisil)imidazolidin-2,4-dione
0,058
10-18
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-methoxybenzyl)imidazolidin-2,4-dione
to 0.060
10-93
1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-propyl-1,2,4-triazolin-3,5-dione
0,062

10-5
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-phenoxyethyl)imidazolidin-2,4-dione
0,063
10-77
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-florfenicol)imidazolidin-2,4-dione
0,064
10-30
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-methylbenzyl)imidazolidin-2,4-dione
0,065
10-94
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-phenylpropyl)imidazolidin-2,4-dione
0,068
10-11
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-hydroxybenzyl)imidazolidin-2,4-dione
0,069

10-6
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-methoxybenzyl)imidazolidin-2,4-dione
0,073
10-23
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-nitrobenzyl)imidazolidin-2,4-dione
of 0.081
10-76
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-methoxyphenethyl)imidazolidin-2,4-dione
0,097
10-13
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-hydroxy-4-methoxybenzyl)imidazolidin-2,4-dione
0,087
10-21
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-terbisil)imidazolidin-2,4-dione
0,090

10-7
methyl 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzoate
is 0.102
10-4
(R)-5-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,112
10-3
(S)-5-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,113
10-2
5-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,117
10-16
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(2-methoxyethoxy)benzyl)imidazolidin-2,4-dione
0,131

10-56
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(3-methoxybenzyl)imidazolidin-2,4-dione
0,131
10-20
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-methylbenzyl)imidazolidin-2,4-dione
of 0.133
10-87
1-(3,4-dimethoxyphenethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,137
10-78
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-fencelineecology-2,4-dione
0,139
10-9
3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)-N-methylbenzamide
0,141

10-85
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-methoxyphenethyl)imidazolidin-2,4-dione
0,158
10-34
1-(2-aminobenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,163
10-24
3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzaldehyde
0,201
10-90
1-((1,3-dimethyl-1H-pyrazole-5-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,215

10-22
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(trifluoromethyl)benzyl)imidazolidin-2,4-dione
0,221
10-75
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((6-methoxypyridine-2-yl)methyl)imidazolidin-2,4-dione
0,229
10-53
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,257
10-81
1-(2-chlorphenyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,248
10-74
1-(cyclohexylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,276

10-58
5-(cyclopentylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,316
10-28
1-((1,5-dimethyl-1H-pyrazole-3-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,322
10-33
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-(hydroxymethyl)benzyl)imidazolidin-2,4-dione
0,322
10-59
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(4-hydroxybenzyl)imidazolidin-2,4-dione
0,327
10-38
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((1-methyl-1H-pyrazole-3-yl)methyl)imidazolidin-2,4-dione
0,363

10-46
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(3-phenylpropyl)imidazolidin-2,4-dione
0,484
10-95
1-benzyl-2-butyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione
0,409
10-17
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(methylsulfinyl)benzyl)imidazolidin-2,4-dione
0,412
10-86
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-hydroxyphenethyl)imidazolidin-2,4-dione
0,412
10-83
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-florfenicol)imidazolidin-2,4-dione
10-55
1-((3,5-dimethylisoxazol-4-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,479
10-60
5-(3,4-dihydroxybenzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,513
10-19
2-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzonitrile
0,550
10-54
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(pyridine-2-ylmethyl)imidazolidin-2,4-dione
0,609
10-71
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((6-(hydroxymethyl)pyridine-2-yl)methyl)imidazolidin-2,4-dione
0,721

10-48
2-(1-(1-((3,5-dim is telesocial-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)-N-phenylacetamide
0,747
10-47
5-(benzoyloxymethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,766
10-96
tert-butyl 3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzylcarbamoyl
0,891
10-61
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-isobutylthiazole-2,4-dione
0,912
10-80
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(2-(naphthalene-1-yl)ethyl)imidazolidin-2,4-dione
0,927

10-57
5-(cyclohexylmethyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,962
10-51
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-Mei is the DIN-2,4-dione
0,966
10-82
1-(3-chlorphenyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,982
10-45
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-fencelineecology-2,4-dione
0,793
10-72
1-((3,4-dimethoxypyridine-2-yl)methyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
0,999

10-25
3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)methyl)benzonitrile
1,003
10-37
1-(3-((dimethylamino)methyl)benzyl)-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
1,220
10-41
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-methylamide alidin-2,4-dione
1,285
10-49
N-benzyl-2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)ndimethylacetamide
1,329
10-73
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-((tetrahydrofuran-2-yl)methyl)imidazolidin-2,4-dione
1,362

10-62
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-isopropylimidazole-2,4-dione
1,440
10-1
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
1,696
10-50
2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)-N-(3-methoxybenzyl)ndimethylacetamide
1,773
10-8
3-((3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,4-dioxoimidazolidin-1-yl)shall ethyl)benzoic acid
1,798
9-5
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidine-2,4,5-Trion
2,493

10-29
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(4-methoxybenzyl)imidazolidin-2,4-dione
3,117
10-63
(E)-5-benzylidene-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2,4-dione
10-67
4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-methyl-2-(2-phenoxyethyl)-1,2,4-triazoline-3,5-dione
10-97
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-(hydroxymethyl)imidazolidin-2,4-dione
9-8
1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethyl-1,3,5-triazine-2,4,6-Trion
6,151

9-7
5-benzyl-1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-3-phenethyl-1,3,5-triazine-2-he
6,580
9-9
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2,4,6-Trion
1,562
10-88
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)imidazolidin-2-he
1,250
9-5
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)tetrahydropyrimidin-2(1H)-he
3,434
9-6
1-benzyl-3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,3,5-triazine-2-he
6,029

/tr>
10-68
4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione
10-45
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-5-fencelineecology-2,4-dione
0,793
10-44
2-(1-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2,5-dioxoimidazolidin-4-yl)acetic acid
10-65
1-benzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-2-methyl-1,2,4-triazolin-3,5-dione
0,019
10-70
1,2-dibenzyl-4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2,4-triazoline-3,5-dione
1,799

10-69
4-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,2-dimethyl-1,2,4-triazolin-3,5-dione
0,707
10-52
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1,5,5-trimethylindolenine-2,4-dione
0,810
10-14
3-(1-((3,5-dimethylisoxazol-4-yl)methyl)-1H-pyrazole-4-yl)-1-(3-(2-methoxyethoxy)benzyl)imidazolidin-2,4-dione
0,071

Example 11. The contribution of hT2R8 in the bitter taste of saccharin in people with insensitive to the taste variants of genes hT2R43 and hT2R44

Figure 6 shows the dependence of the dose-response and the effects of saccharin on the activity of receptors in transfected cells expressing variants hT2R43, hT2R44 and hT2R8. hT2R8 is less sensitive with respect to the saccharin in the analysis ofin vitrothan “sensitive to the taste of alleles hT2R43-W35 and hT2R44-W35, but responds better than “insensitive to taste alleles hT2R43-S35 and hT2R44-R35. Pronin et al.,Curr. Biol.17: 1403-8 (2007). Based on genotypic analysis were selected five subjects with sensitive taste” alleles (hT2R43-W35 and/or hT2R44-W35) and five subjects with “insensitive to the taste” alleles (hT2R43-S35 and hT2R44-R35). Each subject was given 6 pairs of solutions and asked to determine which of the samples in the pair have a more bitter taste. The results, shown below in table 8, indicate that the blocker hT2R8 Cpd-D reduces the bitter taste of saccharin in people with insensitive to the taste” alleles hT2R43 and hT2R44, but does not affect people with sensitive taste” alleles of these genes.

Other typical compounds proposed in the present invention and/or suitable for use in the methods according to the present invention include compounds having the following formulas.

According to the first aspect,the proposed compound of structural formula (I):

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar1represents a five - or six-membered aryl, heteroaryl or cycloalkyl ring;

m is 0, 1, 2 or 3;

R1represents the SO2; O=C; C=S; or C=NOR4;

X is selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R1' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted Alky is, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

or, alternatively, X and/or at least one of R1' together with the atoms to which they are linked, form an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cycloheptadiene or substituted cycloheptatriene ring, and the ring optionally is fused with another aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cyclogeranyl or substituted cyclogeranyl ring;

R4-R8independently selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted g is tetraalkyl, heteroaryl, substituted heteroaryl, heteroaromatic and substituted heteroaromatic, or, alternatively, R5and R6, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring;

A and B are independently selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, and

b is 0, 1 or 2;

According to the second aspect, the invention proposed compounds of structural formula (II)below:

or a salt, hydrate, MES or N-oxides of these compounds, where

Ar1, Ar2and Ar3independently represent a five - or six-membered aryl, heteroaryl or cycloalkyl ring;

m is 0, 1, 2 or 3;

n and p independently are 0, 1, 2, 3 or 4;

r and t are independently 0, 1 or 2;

Y and Z are independently selected from the group including CR6R7, C=O, C=S, C=NOR6, O, NR6and S(O)b;

R1selected from the group comprising the SO2, C=O, C=S and C=NOR4;

X can be selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl,arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, -OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

X is preferably selected from the group comprising hydrogen, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, S(O)bR6, CONR6R7, -CO2R6, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6).

each R1' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2 R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R2' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6) and P(O)(OR5)(OR6);

each R3' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2Rsup> 6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

or, alternatively, X and/or at least one of R1' together with the atoms to which they are linked, form an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cycloheptadiene or substituted cycloheptatriene ring, such ring may be fused with another aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, cyclogeranyl or substituted cyclogeranyl ring;

R4-R8independently represent hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic or, alternatively, R5and R6, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene Kohl is about;

b is 0, 1 or 2.

According to another aspect, the invention proposed compounds having the structural formula (III)below:

or a salt, hydrate, MES or N-oxides of these compounds, where

Ar1, Ar2and Ar3independently represent a five - or six-membered aryl, heteroaryl or cycloalkyl ring, while Ar2and Ar3may be missing;

m is 0, 1, 2 or 3;

n and p independently are 0, 1, 2, 3 or 4;

each R1' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R2' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted aryl is Kil, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R3' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

R5-R8independently represent hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted aryl is Kil, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic or, alternatively, R5and R6, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring;

b is 0, 1 or 2.

According to another aspect, the invention proposed a compound having the structure shown below:

or a salt, hydrate, MES or N-oxide of the specified connection.

According to another aspect, the invention proposed compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds.

According to still other alternative implementations, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds.

According to a related aspect, the proposed compound of structural formula (IV):

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar4and Ar5independently represent a five - or six-membered aryl or heteroaryl ring;

W is selected from the group including the surrounding CR 6R7, C=O, C=S; C=NOR6O, NR6, S, SO, SO2and (CH2)n;

n is 0, 1, 2 or 3;

G is selected from the group including CR6R7, C=O, C=S, C=NOR6and S(O)b;

R20selected from the group comprising hydrogen, arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R21selected from the group comprising arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R6and R7independently selected from the group comprising hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic, or, alternatively, R6and R7together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring; and

b is 0, 1 or 2.

According to yet another related aspect, the proposed compound of structural formula (V):

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar4and Ar5independently represent a p is t - or six-membered aryl or heteroaryl ring;

n is 0, 1, 2 or 3;

R21selected from the group comprising arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R35selected from the group comprising hydrogen, alkyl and substituted alkyl.

According to an additional variant of the invention, the proposed compound of structural formula (VI)

or a salt, hydrate, MES or N-oxide of the compounds, where

R30selected from the group comprising arylalkyl, heteroaromatic, arylalkyl, heteroaromatic, aryl, heteroaryl, alkyl, alkoxy-alkyl, alkenyl, cycloalkyl, cycloalkenyl and substituted derivatives;

R35selected from the group comprising hydrogen, alkyl and substituted alkyl.

According to an additional variant of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds,

where each R independently represents Cl, MeO, CN, EtO, HE, Me, -SO2Me, F or N, and

n is 0, 1, 2, 3, or 4.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or oxide of these compounds, where each R independently represents a MeO or HE, and n is 0, 1, 2, 3, or 4.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds, where R represents H, Me, Et, OCOMe, CH2HE or Ph OMe.

According to the following variants of implementation, the invention

the proposed compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds. According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds. According to other variants of implementation, the proposed invention compounds having the structure shown below:

or a salt, hydrate, MES or N-oxides of these compounds.

According to other variants of implementation, the proposed invention compounds having the structure shown below:

According to one aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or proletar the TSS specified connection,

where Ar6and Ar7that may be the same or different and independently from each other, represent a five - or six-membered aryl group or a five - or six-membered heteroaryl group;

Alk represents an alkyl group, optionally containing a heteroatom;

R36and R37that may be the same or different independently of one another, represent H, alkyl, or R36and R37together with the atoms to which they are attached, form an optionally substituted five - or six-membered heterocycle; and

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated.

According to one aspect, the compounds proposed in the invention contain the five-membered heterocycle. According to one implementation variant, the five-membered heterocycle is an as or a substituted or unsubstituted cyclic urea.

According to the SNO one implementation variant, the as is a as the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Ar6and Ar7that may be the same or different and independently from each other, represent a five - or six-membered aryl group or a five - or six-membered heteroaryl group;

Alk represents an alkyl group, optionally containing a heteroatom;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated; and

R39and R40that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl alkyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted aryl shall kilmichael, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated, or R39and R40together with the carbon atom to which they are attached, form a C=O group, or a substituted or unsubstituted alkenylphenol group.

According to another aspect, the compounds proposed in the invention contain the five-membered heterocycle, which is orasol. According to one implementation variant, the specified orasol is orasol formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Ar6and Ar7that may be the same or different and independently from each other, represent a five - or six-membered aryl group or a five - or six-membered heteroaryl group;

Alk represents an alkyl group, optionally containing a heteroatom;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or nezame the military arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated; and

R41represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated.

According to another aspect, the compounds proposed in the invention contain a six-membered heterocycle. According to one implementation variant, six-membered heterocycle represents a six-membered heterocycle of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or n is substituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated; and

R42, R43, R44, R45and R46that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl alkyl, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, or R42and R43or R45and R46together with the carbon atoms to which they are both attached, form a C=O group.

According to another aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50predstavljaet a H or substituted or unsubstituted alkyl;

R36and R37that may be the same or different independently of one another, represent H, alkyl, or R36and R37together with the atoms to which they are attached, form an optionally substituted five - or six-membered heterocycle; and

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen.

According to another aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or Strait is carstvo specified connection,

where Alk is an alkyl group, optionally containing a heteroatom;

T1represents the C=O and Q represents CR51R52or NR51where R51and R52that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated, or R51and R52together with the carbon atom to which they are attached, form a C=O group, or a substituted or unsubstituted alkenylphenol group;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50represents H or substituted or unsubstituted alkyl;

R38represents H, substituted or nezamedin the th alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen.

According to another aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds.

According to another aspect, the invention relates to a compound of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

T2represents the C=S, C=O or S(O)2;

R53represents a substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted aryl or substituted or unsubstituted arylalkyl;

M1represents N or CR54where R54represents H or substituted or unsubstituted alkyl;

M2represents N or CR55where R55represents H or substituted or unsubstituted alkyl;

R56represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R57represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen;

this decree is effective method involves reacting a compound of the formula:

where R56, R57and Alk are defined above, and J is a leaving group;

with the compound of the formula:

where M1and M2defined above, to obtain compounds of formula

,

contains NO2group;

the restoration of NO2group to obtain compounds containing NH2group; and

the interaction of compounds containing NH2the group, with the compound of the formula

where J2represents a leaving group, and T2and R53defined above.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

R51and R52that may be the same or different independently of one another, represent H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted the first or the unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated, or R51and R52together with the carbon atom to which they are attached, form a substituted or unsubstituted alkenylphenol group;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50represents H or substituted or unsubstituted alkyl;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen;

while this method involves the heating of the compounds of formula:

where R47, R48, Alk, M1and M2defined above;

for turning-CON3group-N=C=O group, and the subsequent interaction with the compound of the formula:

where J3represents a leaving group, and R38, R51and R52defined above.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or a salt, hydrate, MES, N-oxide or prodrug of the compounds,

where Alk is an alkyl group, optionally containing a heteroatom;

R52represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted arylalkylamines, substituted or unsubstituted heteroaromatic, substituted or unsubstituted heteroarylboronic, for ewenny or an unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic, halogenated;

M1represents N or CR49where R49represents H or substituted or unsubstituted alkyl;

M2represents N or CR50where R50represents H or substituted or unsubstituted alkyl;

R38represents H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted geterotsiklicheskikh, substituted or unsubstituted aryl, substituted or unsubstituted arylamidase, substituted or unsubstituted heteroaromatic, substituted or unsubstituted arylalkyl, substituted or unsubstituted, Allakaket, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroaromatic or halogenated;

R47represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen; and

R48represents H, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl or halogen;

while this method includes the Naga is evanie compounds of the formula:

where R47, R48, Alk, M1and M2defined above;

for turning-CON3group-N=C=O group and the subsequent interaction with a hydrazine of the formula:

where R38defined above.

According to another aspect, the invention relates to a method for obtaining compounds of the formula:

or a salt, hydrate, MES or N-oxide of the compounds, where

Ar1, Ar2and Ar3independently represent a five - or six-membered aryl, heteroaryl or cycloalkyl ring, while Ar2and Ar3may be missing;

m is 0, 1, 2 or 3;

n and p independently are 0, 1, 2, 3 or 4;

each R1' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7 B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R2' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

each R3' are independently selected from the group comprising hydrogen, halogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, acyl, substituted acyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic, substituted heteroaromatic, CN, NO2, OR6, S(O)bR6, NR6R7, CONR6R7, CO2R6, NR6CO2R7, NR6CONR7R8, NR6CSNR7R8, NR6C(=NH)NR7R8, SO2NR5R6, NR5SO2R6, NR5SO2 NR6R7B(OR5)(OR6), P(O)(OR5)(OR6) and P(O)(R5)(OR6);

R5-R8independently represent hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, heteroaromatic or substituted heteroaromatic or, alternatively, R6and R7, R7and R8together with the atoms to which they are linked, form cycloheptadiene or substituted cycloheptatriene ring;

b is 0, 1 or 2;

while this method involves reacting a compound of the formula:

where J is a leaving group;

with the compound of the formula:

with the receipt of the product; and

the interaction of the specified product with the compound of the formula:

where J is a leaving group.

The SEQUENCE of the GENES AND POLYPEPTIDES of the CHIMERIC G-PROTEIN AND HT2R

The sequence of the protein rodopoulou tags: (SEQ ID NO:1)

MNGTEGPNFYVPFSNKTGVVRSPFEAPQYYLAEPW

The sequence of the protein G16gust44: (SEQ ID NO:2)

MARSLTWRCCPWCLTEDEKAAARVDQEINRILLEQKKQDRGELKLLLLGPGESGKSTFIKQMRIIHGAGYSEEERKGFRPLVYQNIFVSMRAMIEAMERLQIPFSRPESKHHASLVMSQDPYKVTTFEKRYAAAMQWLWRDAGIRACYERRREFHLLDSAVYYLSHLERITEEGYVPTAQDVLRSRMPTTGINEYCFSVQKTNLRIVDVGGQKSERKKWIHCFENVIALIYLASLSEYDQCLEENNQENRMKESLALFGTILELPWFKSTSVILFLNKTDILEEKIPTSHLATYFPSFQGPQDAEAAKRFILDMYTRMYTGCVDGPEGSNLKKEDKEIYSHMTCATDTQNVKFVFDAVTDIIIKENLKDCGLF

Sequence hT2R8:

DNA(SEQ ID NO:3) ATGTTCAGTCCTGCAGATAACATCTTTATAATCCTAATAACTGGAGAATTCATACTAGGAATATTGGGGAATGGATACAT

TGCACTAGTCAACTGGATTGACTGGATTAAGAAGAAAAAGATTTCCACAGTTGACTACATCCTTACCAATTTAGTTATCG

CCAGAATTTGTTTGATCAGTGTAATGGTTGTAAATGGCATTGTAATAGTACTGAACCCAGATGTTTATACAAAAAATAAA

CAACAGATAGTCATTTTTACCTTCTGGACATTTGCCAACTACTTAAATATGTGGATTACCACCTGCCTTAATGTCTTCTA

TTTTCTGAAGATAGCCAGTTCCTCTCATCCACTTTTTCTCTGGCTGAAGTGGAAAATTGATATGGTGGTGCACTGGATCC

TGCTGGGATGCTTTGCCATTTCCTTGTTGGTCAGCCTTATAGCAGCAATAGTACTGAGTTGTGATTATAGGTTTCATGCA

ATTGCCAAACATAAAAGAAACATTACTGAAATgttccatgtgagtaaaataccatactttgaacccttaactctctttaa

CCTGTTTGCAATTGTCCCATTTATTGTGTCACTGATATCATTTTTCCTTTTAGTAAGATCTTTATGGAGACATACCAAGC

AAATAAAACTCTATGCTACCGGCAGTAGAGACCCCAGCACAGAAGTTCATGTGAGAGCCATTAAAACTATGACTTCATTT

ATCTTCTTTTTTTTCCTATACTATATTTCTTCTATTTTGATGACCTTTAGCTATCTTATGACAAAATACAAGTTAGCTGT

GGAGTTTGGAGAGATTGCAGCAATTCTCTACCCCTTGGGTCACTCACTTATTTTAATTGTTTTAAATAATAAACTGAGGC

AGACATTTGTCAGAATGCTGACATGTAGAAAAATTGCCTGCATGATATGA

Protein(SEQ ID NO:4)

MFSPADNIFIILITGEFILGILGNGYIALVNWIDWIKKKKISTVDYILTNLVIARICLISVMVVNGIVIVLNPDVYTKNK

QQIVIFTFWTFANYLNMWITTCLNVFYFLKIASSSHPLFLWLKWKIDMVVHWILLGCFAISLLVSLIAAIVLSCDYRFHA

IAKHKRNITEMFHVSKIPYFEPLTLFNLFAIVPFIVSLISFFLLVRSLWRHTKQIKLYATGSRDPSTEVHVRAIKTMTSF

IFFFFLYYISSILMTFSYLMTKYKLAVEFGEIAAILYPLGHSLILIVLNNKLRQTFVRMLTCRKIACMI

Sequence hT2R14:

DNA(SEQ ID NO:5) ATGGGTGGTGTCATAAAGAGCATATTTACATTCGTTTTAATTGTGGAATTTATAATTGGAAATTTAGGAAATAGTTTCAT

AGCACTGGTGAACTGTATTGACTGGGTCAAGGGAAGAAAGATCTCTTCGGTTGATCGGATCCTCACTGCTTTGGCAATCT

CTCGAATTAGCCTGGTTTGGTTAATATTCGGAAGCTGGTGTGTGTCTGTGTTTTTCCCAGCTTTATTTGCCACTGAAAAA

ATGTTCAGAATGCTTACTAATATCTGGACAGTGATCAATCATTTTAGTGTCTGGTTAGCTACAGGCCTCGGTACTTTTTA

TTTTCTCAAGATAGCCAATTTTTCTAACTCTAtttttctctacctaaagtggagagttaaaaaggtggttttggtgctgc

TTCTTGTGACTTCGGTCTTCTTGTTTTTAAATATTGCACTGATAAACATCCATATAAATGCCAGTATCAATGGATACAGA

AGAAACAAGACTTGCAGTTCTGATTCAAGTAACTTTACACGATTTTCCAGTCTTATTGTATTAACCAGCACTGTGTTCAT

TTTCATACCCTTTACTTTGTCCCTGGCAATGTTTCTTCTCCTCATCTTCTCCATGTGGAAACATCGCAAGAAGATGCAGC

ACACTGTCAAAATATCCGGAGACGCCAGCACCAAAGCCCACAGAGGAGTTAAAAGTGTGATCACTTTCTTCCTACTCTAT

GCCATTTTCTCTCTGTCTTTTTTCATATCAGTTGGACCTCTGAAAGGTTGGAGGAAAATCTAATTATTCTTTCCCAGGT

GATGGGAATGGCTTATCCTTCATGTCACTCATGTGTTCTGATTCTTGGAAACAAGAAGCTGAGACAGGCCTCTCTGTCAG

TGCTACTGTGGCTGAGGTACATGTTCAAAGATGGGGAGCCCTCAGGTCACAAAGAATTTAGAGAATCATCTTGA

Protein(SEQ ID NO:6)

MGGVIKSIFTFVLIVEFIIGNLGNSFIALVNCIDWVKGRKISSVDRILTALAISRISLVWLIFGSWCVSVFFPALFATEK

MFRMLTNIWTVINHFSVWLATGLGTFYFLKIANFSNSIFLYLKWRVKKVVLVLLLVTSVFLFLNIALINIHINASINGYR

RNKTCSSDSSNFTRFSSLIVLTSTVFIFIPFTLSLAMFLLLIFSMWKHRKKMQHTVKISGDASTKAHRGVKSVITFFLLY

AIFSLSFFISVWTSERLEENLIILSQVMGMAYPSCHSCVLILGNKKLRQASLSVLLWLRYMFKDGEPSGHKEFRESS

Although in the above detailed description discloses several embodiments of the present invention, it should be understood that this description is illustrative only and not limiting of the disclosed invention. The present invention is limited only by the formula below.

1. The compound of structural Formula (I)below:
the compound of the formula:

or its pharmaceutically acceptable salt,
where Alk represents a C1-C6alkyl group;
G represents C=O and Q represents CR51R52or NR51where R51and R52being the same or different, independently of one another, represent H, C1-C6alkyl, optionally substituted Deputy selected from the group comprising carboxy, phenoxy, benzyloxy, C1-C6alkoxy and hydroxy; C3-C6cycloalkyl1-C6alkyl; phenyls1-C6alkyl, optionally substituted with halogen; phenylamide the 1-C6alkyl; phenyls1-C6alkylamides1-C6alkyl, optionally substituted C1-C6alkoxygroup;
or R51and R52together with the carbon atom to which they are attached, form the group C=O or C2-C6alkenylphenol group, optionally substituted by phenyl;
M1represents CR49where R49represents H;
M2represents CR50where R50represents H;
R38represents H, C1-C6alkyl, substituted phenoxypropane; C3-C6cycloalkyl1-C6alkyl; arils1-C6alkyl, optionally substituted by 1 or 2 substituents selected from the group comprising From1-C6alkyl, C1-C6alkoxy, C1-C6alkoxycarbonyl, carboxyl, N-methylamino, hydroxy, C1-C6alkoxyl1-C6alkoxy, C1-C6alkylthio,1-C6alkylsulfonyl, cyano, halogen, perfors1-C6alkyl, nitro, formyl, hydroxys1-C6alkyl and amino, and aryl fragment represents a phenyl or naphthyl; and heteroaryl1-C6alkyl, where the heteroaryl fragment represents pyridinyl, optionally substituted 1 or 2 groups selected from C1-C6alkoxy or guide who oxis 1-C6of alkyl, pyrazolyl or isoxazolyl, substituted by 1 or 2 C1-C6alkyl groups;
R47represents a C1-C6alkyl; and
R48represents a C1-C6alkyl.

2. The compound having the formula:

or its pharmaceutically acceptable salt.

3. The compound having the formula:


or its pharmaceutically acceptable salt

4. The compound having the formula:

or its pharmaceutically acceptable salt.

5. The compound having the formula:


or its pharmaceutically acceptable salt,
where each R independently represents Cl, MeO, CN, EtO, OH, Me, -SO2Me, F, H, and
n is 0, 1, 2, 3, or 4.

6. The compound having the formula:

or its pharmaceutically acceptable salt,
where each R independently represents MeO, OH, and
n is 0, 1, 2, 3, or 4.

7. The compound having the formula:



or its pharmaceutically acceptable salt,
where R is selected from the group consisting of H, Me, Et, OCOMe, CH2OH, OMe and Ph.

8. The compound having the formula:





or its pharmaceutically acceptable salt.

9. The compound having the formula:



or its pharmaceutically acceptable salt.

10. The compound having the formula:



or its pharmaceutically acceptable salt,
where Adamantyl means of substituted.

11. The compound having the formula:

or its pharmaceutically acceptable salt.

12. The compound having the formula:























or its pharmaceutically acceptable salt.

13. The compound having the formula:





































































or its pharmaceutically acceptable salt.

14. The compound having the formula:









or its pharmaceutically acceptable salt.

15. The compound according to claim 1, having the formula:



























or its pharmaceutically acceptable salt.

16. The compound according to claim 1, having the formula:





or its pharmaceutically acceptable salt.

17. The compound having the formula:

or its pharmaceutically priemel who may salt.

18. A method of reducing or diminishing the bitter taste, which includes adding at least one compound according to one of claims 1 to 17 or similar to the composition for ingestion by humans or animals in a concentration effective to alleviate or reduce the bitter taste associated with this song.

19. The method according to p, in which the concentration of at least one of the compounds is from about 0.1 million parts to about 100 million parts.

20. The method according to p, in which the concentration of at least one of the compounds is from about 1 million to about 25 million parts.

21. The method according to p, in which the composition is a food product, beverage, or medicinal product.

22. The method according to p, in which the composition contains bitter compounds that activate one or more of the hT2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 65, 71 and/or hT2R5, 9, 13, 54, 67 and 75.

23. The method according to p, in which the composition contains at least one bitter compound, which activates multiple bitter taste receptors, or which has an undetermined specificity to receptors bitter taste.

24. The method according to p, in which the composition is a coffee or a composition of the flavored coffee.

25. The method according to paragraph 24, in which the composition contains at least 2 compounds, at least one of the C which is an antagonist of hT2R8 and at least one of which is an antagonist hT2R14.

26. The method according to paragraph 24, in which coffee or flavored coffee composition is an instant coffee or brewed coffee.

27. The composition of the food product or beverage or composition of the medicinal product or composition of non-food product that contains at least one compound according to one of claims 1 to 17 to reduce or mitigate the bitter taste.

28. The composition of the food product or beverage, or composition of the medicinal product, or composition of non-food product according to item 27, where the composition is a food product or beverage is a coffee or flavored coffee composition of the food or drink.

29. The composition of the food product or beverage, or composition of the medicinal product, or composition of non-food product according to item 27, where the composition is a food product or beverage or composition of the medicinal product or composition of food product contains at least 2 compounds, at least one of which is an antagonist of hT2R8 and at least one of which is an antagonist hT2R14.

30. The composition of the food product or beverage, or composition of the medicinal product, or composition of non-food product according to item 27, where the composition of the drink is a ready-to-use soluble and dry coffee drink, a mixture of coffee nab the TKA or concentrate coffee drink.

31. The composition of the food product or beverage, or composition of the medicinal product, or composition of non-food product according to item 27, where the composition of the drink is a substitute for cream based on milk, non-dairy substitute cream or samelevel for coffee drinks.

32. The composition of the food product or beverage, or composition of the medicinal product, or composition of non-food product according to item 27, where the composition of the food product is a Supplement, nutraceuticals, functional food, pharmaceutical drug, pharmaceutical drug, over-the-counter product for the care of oral hygiene or beauty product.

33. The composition of the food product or beverage, or composition of the medicinal product, or composition of food product p, where the product is to care for the oral cavity is a tool for the care of teeth, the liquid mouthwash or chewing gum.

34. The method of detection or quantitative analysis of the possible block the bitter taste of bitter compounds in the composition of the compound according to one of claims 1 to 17, and the method includes
contacting a composition containing at least one bitter compound, which activates at least one human bitter taste receptor selected and the group, consisting of hT2R3, 7, 10, 14, 16, 44, 51, 55, 61, 63, 64, 65 or 71 and/or hT2R5, 9, 13, 54, 67 and 75, and
identifying whether the activation of the specified at least one receptor is blocked or inhibited by the connection according to one of claims 1 to 17, where the specified blocking or inhibition reduces the bitter taste of at least one bitter compound.

35. The method according to clause 34, further comprising a taste test in which a bitter connection and the connection specified antagonist taste separately and in combination, to confirm that bitter taste weak antagonist.

36. The method according to clause 34, which is a high-performance analysis.

37. The method of obtaining the compounds of formula:

or its pharmaceutically acceptable salt,
where Alk represents a C1-C6alkyl group;
R51and R52being the same or different, independently of one another, represent H, C1-C6alkyl, optionally substituted Deputy selected from the group comprising carboxy, phenoxy, benzyloxy, C1-C6alkoxy and hydroxy;
C3-C6cycloalkyl1-C6alkyl; phenyls1-C6alkyl, optionally substituted by one Deputy, selected from the group comprising halogen; phenylamides1-C6alkyl; phenyls1 -C6alkylamides1-C6alkyl, optionally substituted C1-C6alkoxygroup;
or R51and R52together with the carbon atom to which they are attached, form the group C=O or C2-C6alkenylphenol group, optionally substituted by phenyl;
M1represents CR49where R49represents H;
M2represents CR50where R50represents H;
R38represents H, C1-C6alkyl, substituted phenoxypropane; C3-C6alcauciles1-C6alkyl; arils1-C6alkyl, optionally substituted by 1 or 2 substituents selected from the group comprising From1-C6alkyl, C1-C6alkoxy, C1-C6alkoxycarbonyl, carboxyl, N-methylamino, hydroxy, C1-C6alkoxyl1-C6alkoxy, C1-C6alkylthio, C1-C6alkylsulfonyl, cyano, halogen, perfors1-C6alkyl, nitro, formyl, hydroxys1-C6alkyl and amino, and aryl fragment represents a phenyl or naphthyl; and heteroaryl1-C6alkyl, where the heteroaryl fragment represents pyridinyl, optionally substituted 1 or 2 groups selected from C1-C6alkoxy or hydraxis1-C6of alkyl, pyrazole is or isoxazolyl, substituted by 1 or 2 C1-C6alkyl groups;
R47represents a C1-C6alkyl; and
R48represents a C1-C6alkyl;
where the method includes heating the compounds of formula:

where R47, R48, Alk, M1and M2defined above;
for group conversion-CON3group-N=C=O and subsequent interaction with the compound of the formula:

where J3represents a leaving group, and R38, R51and R52defined above.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of structural formula or a salt thereof, where each of Z1, Z2 and Z3 is independently selected from N and C(R9), where not more than one of Z1, Z2 and Z3 is N; each R9 is hydrogen; and is a second chemical bond between either W2 and C(R12), or W1 and C(R12); W1 is -N=, and W2(R14) is selected from -N(R14)- and -C(R14)=, such that when W1 is -N=, W2(R14) is -N(R14)- and is a second chemical bond between W1 and C(R12); R11 is selected from phenyl and a heterocycle which is selected from a saturated or aromatic 5-6-member monocyclic ring, which contains one or two or three heteroatoms selected from N, O and S, or an 8-member bicyclic ring which contains one or more heteroatoms selected from N, O and S, where R11 is optionally substituted with one or two substitutes independently selected from halogen, C1-C4 alkyl, =O, -O-R13, -(C1-C4 alkyl)-N(R13)(R13), -N(R13)(R13), where each R13 is independently selected from -C1-C4alkyl; or two R13 together with a nitrogen atom to which they are bonded form a 5-6-member saturated heterocycle, optionally containing an additional heteroatom selected from NH and O, where if R13 is an alkyl, the alkyl is optionally substituted with one or more substitutes selected from -OH, fluorine, and if two R13 together with the nitrogen atom to which they are bonded form a 5-6-member saturated heterocycle, the saturated heterocycle is optionally substituted on any carbon atom with fluorine; R12 is selected from phenyl, a 4-6-member monocyclic saturated ring and a heterocycle, which is selected from an aromatic 5-6-member monocyclic ring which contains one or two heteroatoms selected from N and S, where R12 is optionally substituted with one or more substitutes independently selected from halogen, -C≡N, C1-C4 alkyl, C1-C2 fluorine-substituted alkyl, -O-R13, -S(O)2-R13, -(C1-C4 alkyl)-N(R13)(R13), -N(R13)(R13); R14 is selected from hydrogen, C1-C4 alkyl, C1-C4 fluorine-substituted alkyl, C1-C4 alkyl-N(R13)(R13), C1-C4 alkyl-C(O)-N(R13)(R13); and X1 is selected from -NH-C(=O)-†, -C(=O)-NH-†, -NH-S(=O)2-†, where † denotes the point where X1 is bonded to R11. The invention also relates to a pharmaceutical composition having sirtuin modelling activity based on said compounds.

EFFECT: obtaining novel compounds and a pharmaceutical composition based on said compounds, which can be used in medicine to treat a subject suffering from or susceptible to insulin resistance, metabolic syndrome, diabetes or complications thereof.

18 cl, 2 tbl, 52 ex

Organic compounds // 2518462

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula

and

,

where X represents S or O, one of X1 and X2 represents CR3' and second represents N or independently CR3', n represents integer number 1, 2 or 3; R1 represents C1-6 halogenalkyl, R2 is selected from halogen and C1-C6-halogenalkyl; R3' represents H, C1-C6-alkyl, halogen, cyanogroup, or phenyl, non-substituted or substituted with halogen, C1-C6-alcoxygroup, C1-C6-halogenalcoxygroup, C1-C6-halogenalkyl group; Z represents halogen, Q radical or group -C(O)-NR5R6; R5 represents H or C1-C4-alkyl, R6 represents H; Q', C1-C6-alkyl, non-substituted or substituted with halogen, cyanogroup, C1-C4-alcoxygroup, C1-C4-alkoxycarbonyl, C2-C4-alkanoyl, aminocarbonyl, N-mono- or N,N-di-C1-C2-alkylaminocarbonyl, C1-C4-alkylthiogroup, group -C(O)NHR7 or radical Q"; or C3-C6-cycloalkyl, substituted with group -C(O)NHR7; or C2-C4-alkinyl; Q, Q' and Q" are such as given in the invention formula; R7 represents C1-C6-alkyl, which is non-substituted or substituted with halogen, cyanogroup, pyridyl; or represents C2-C4-alkinyl. Invention also relates to composition for fighting ectoparasites, containing compound of formula (Ia) or (Ib), and to application of compounds of formula (Ia) or (Ib) for composition production.

EFFECT: compounds of formula (Ia) and (Ib), possessing activity against ectoparasites.

11 cl, 4 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula (I), wherein R1 represents an alkoxy group or halogen; each U and V independently represents CH or N; "----" means a bond or is absent; W represents CH or N, or if "----" is absent, then W represents CH2 or NH, provided not all U, V and W represent N; A represents a bond or CH2; R2 represents H, or provided A means CH2, then it also can represent OH; each m and n are independently equal to 0 or 1; D represents CH2 or a bond; G represents a phenyl group that is single or double substituted in meta- and/or para-position(s) by substitutes specified in alkyl, C1-3alkoxy group and halogen, or G represents one of the groups G1 and G2: wherein each Z1, Z2 and Z3 represents CH; and X represents N or CH and Q represents O or S; it should be noted that provided each m and n are equal to 0, then A represents CH2; or a pharmaceutically acceptable salt of such compound. Besides, the invention refers to a pharmaceutical composition for treating a bacterial infection containing an active ingredient presented by a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one therapeutically inert additive.

EFFECT: preparing the oxazolidine compounds applicable for preparing a drug for treating and preventing the bacterial infections.

14 cl, 8 dwg, 2 tbl, 33 ex

Cetp inhibitors // 2513107

FIELD: chemistry.

SUBSTANCE: invention relates to compound of formula I, or its pharmaceutically acceptable salt where: X stands for -O-; Z stands for -C(=O)-; Y stands for -(CRR1)-, where R1 is selected from -C1-C2alkyl; R stands for H or -C1-C5alkyl; R5 stands for H; R2 and B each is selected from A1 and A2, where one of R2 and B stands for A1, and the other from R2 and B stands for A2; where A1 has structure (a); A2 is selected from the group, which includes phenyl, pyridyl, pyrazolyl, thienyl, 1,2,4-triazolyl and imodazolyl; A3 is selected from the group including phenyl, thiazolyl and pyrazolyl; A4 is selected from the group, including phenyl, pyridyl, thiazolyl, pyrazolyl, 1,2,4-triazolyl, pyrimidinyl, piperidinyl, pyrrolidinyl and asetidinyl; A2 is optionally substituted with 1-3 substituents, independently selected from halogen atom, -OCH3 and -OCF3 and -C1-C3alkyl, optionally substituted with 1-3 halogen atoms; A3 is substituted with one A4 group and is optionally substituted with 1-2 substituents, independently selected from halogen atom, -OH, -OCH3, -OCF3 and -C1-C3alkyl, optionally substituted with 1-3 halogen atoms; A4 is optionally substituted with 1-3 substituents, independently selected from the group, which includes: (a) -C1-C5alkyl, optionally substituted with 1-3 halogen atoms and optionally substituted with group -OH, (b) -C2-C4alkenyl, optionally substituted with 1-3 halogen atoms, (c) -C(=O)C1-C2alkyl, optionally substituted with 1-3 halogen atoms and optionally substituted with one group selected from -OH, -CO2CH3, -C(=O)CH3, -NR3R4 and -OC1-C2alkyleneOC1-C2alkyl, (d) -C(=O)H, (e) -CO2H, (f) -CO2C1-C4alkyl, optionally substituted with one group, selected from -C(=O)C1-C2alkyl, -OH, -CO2CH3, -CO2H, -NR3R4 and -OC1-C2alkyleneOC1-C2alkyl, (g) -OH, (h) -S(O)xC1-C2alkyl, (i) halogen atom, (j) -CN, (k) -NO2, (l) -C(=O)NR3R4, (m) -OC1-C2alkyleneOC1-C2alkyl, (n) -OC1-C3alkyl, optionally substituted with 1-3 halogen atoms, (o) -C(=O)OC1-C2alkyl, optionally substituted with 1-3 halogen atoms and optionally substituted with one group, selected from -OH, -CO2CH3, -NR3R4 and -OC1-C2alkyleneOC1-C2alkyl, (q) -NR3R4 and (r) -S(O)xNR3R4, on condition that A4 stands for heterocyclic group, attached to A3 by means of ring carbon atom in A4, at least, one substituent in A4 must be selected from Re, where Re is selected from the group including: (a) -C1-C5alkyl, substituted with -OH group and optionally substituted with 1-3 halogen atoms, (b) -C2-C4alkenyl, optionally substituted with 1-3 halogen atoms, (c) -C(=O)C1-C2alkyl, optionally substituted with 1-3 halogen atoms and optionally substituted with one group selected from -OH, -CO2CH3, -C(=O)CH3, -NR3R4 and -OC1-C2alkyleneOC1-C2alkyl, (d) -C(=O)H, (e) -CO2H, (f) -CO2C1-C4alkyl, optionally substituted with one group, selected from -C(=O)C1-C2alkyl, -OH, -CO2CH3, -CO2H, -NR3R4 and -OC1-C2alkyleneOC1-C2alkyl, (g) -OH, (h) -S(O)xC1-C2alkyl, (i) -CN, (j) -NO2, (k) -C(=O)NR3R4, (l) -OC1-C2alkyleneOC1-C2alkyl, (m) -C(=O)C1-C2alkyl, optionally substituted with 1-3 halogen atoms and optionally substituted with one group, selected from -OH, -CO2CH3, -NR3R4 and -OC1-C2alkyleneOC1-C2alkyl, (n) -NR3R4(=O)OC1-C2alkyl, (o) -NR3R4 and (p) -S(O)xNR3R4; p equals 0, 1 or 2; and Ra is selected from halogen atom, -CH3, -CF3, -OCH3 and -OCF3; R3 and R4 each is independently selected from H and CH3; and x equals 0, 1 or 2.

EFFECT: formula (I) compound is applied for medication, which possesses properties of CETP inhibitor, for increase of HDL-C and for reduction of LDL-C Technical result is compounds, inhibiting cholesterol ether transferring protein (CETP).

10 cl, 140 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula (I) , where A is a 6-member heteroaryl, having 1 nitrogen atom as a heteroatom, substituted with 2-3 substitutes such as indicated in the claim, R5 is a halogen atom, cyano or C1-C6alkyl, optionally substituted with a halogen atom; R6 is C1-C6 alkyl, optionally substituted with OH; C1-C3 alkenyl; a 5-member heteroaryl, having 2-4 heteroatoms, each independently selected from N, O or S, substituted with 0-2 substitutes such as indicated in the claim, R10 is a 5-member heteroaryl, having 2-3 heteroatoms, each selected from N, O or S, substituted with 0-2 substitutes, which are C1-C3 alkyl; R7, R8, R17 denote a hydrogen or halogen atom. The invention also relates to a pharmaceutical composition, having BK B2 receptor inhibiting activity, which contains compounds of formula (I), a method of inhibiting, a method of localising or detecting the BK B2 receptor in tissue, use of the compounds of compositions to produce a medicinal agent and methods for treatment.

EFFECT: compounds of formula (I) as BK B2 receptor inhibitors.

22 cl, 1 tbl, 54 ex

FIELD: biotechnologies.

SUBSTANCE: invention refers to a compound of formula (I):

,

where R1 represents NR7C(O)R8 or NR9R10; R2 represents hydrogen; R3 represents halogen; R4 represents hydrogen, halogen, cyano, hydroxy, C1-4alkyl, C1-4alkoxy, CF3, OCF3, C1-4alkylthio, S(O)(C1-4alkyl), S(O)2(C1-4alkyl), CO2H or CO2(C1-4alkyl); R5 represents C1-6alkyl (replaced with NR11R12 or heterocyclyl that represents nonaromatic 5-7-membered ring containing 1 or 2 heteroatoms independently chosen from a group containing nitrogen, oxygen or sulphur); R6 represents hydrogen, halogen, hydroxy, C1-4alkoxy, CO2H or C1-6alkyl (possibly replaced with NR15R16 group, morpholinyl or thiomorpholinyl); R7 represents hydrogen; R8 represents C3-6cycloalkyl (possibly replaced with NR24R25 group), phenyl or heteroaryl, which represents aromatic 5- or 6-membered ring containing 1 to 3 heteroatoms independently chosen from the group containing nitrogen, oxygen and sulphur, and which is probably condensed with one 6-membered aromatic or nonaromatic carbocyclic ring or with one 6-membered aromatic heterocyclic ring, where the above 6-membered aromatic heterocyclic ring includes 1 to 3 heteroatoms independently chosen from a group containing nitrogen, oxygen and sulphur; R9 represents hydrogen or C1-6alkyl (possibly replaced with pyrazolyl); R10 represents C1-6alkyl (possibly replaced with phenyl or heteroaryl group, which represents aromatic 5- or 6-membered ring containing 1 or 2 heteroatoms independently chosen from the group containing nitrogen, oxygen or sulphur, and which is possibly condensed with one 6-membered heterocyclic ring, where the above 6-membered aromatic heterocyclic ring contains 1 or 2 heteroatoms independently chosen from the group containing nitrogen, oxygen or sulphur; where the above phenyl and heteroaryl groups in R8, R9 and R10 are possibly independently replaced with the following group: halogen, hydroxy, C(O)R42, C1-6alkyl, C1-6hydroxyalkyl, C1-6halogenoalkyl, C1-6alkoxy(C1-6)alkyl or C3-10cycloalkyl; unless otherwise stated, heterocyclyl is possibly replaced with group of C1-6alkyl, (C1-6alkyl)OH, (C1-6alkyl)C(O)NR51R52 or pyrrolidinyl; R42 represents C1-6alkyl; R12, R15 and R25 independently represent C1-6alkyl (possibly replaced with hydroxy or NR55R56 group); R11, R16, R24, R51, R52, R55 and R56 independently represent hydrogen or C1-6alkyl; or to its pharmaceutically acceptable salts.

EFFECT: new compounds are obtained, which can be used in medicine for treatment of PDE4-mediated disease state.

10 cl, 2 tbl, 202 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to organic chemistry and specifically to 5-phenyl-1H-pyrazin-2-one derivatives of general formula II or pharmaceutically acceptable salts thereof, where R denotes -R1 or - R1-R2-R3; R1 denotes aryl or heteroaryl, and is optionally substituted with one or two R1'; where each R1' independently denotes C1-6alkyl, halogen or C1-6halogenalkyl; R2 denotes -C(=O), -CH2-; R3 denotes R4; where R4 denotes an amino group or heterocycloalkyl, and is optionally substituted with one or two substitutes selected from C1-6alkyl, hydroxy group, oxo group, C1-6hydroxyalkyl, C1-6alkoxy group; Q denotes CH2; Y1 denotes C1-6alkyl; Y2 denotes Y2b; where Y2b denotes C1-6alkyl, optionally substituted with one Y2b'; where Y2b' denotes a hydroxy group, n and m are equal to 0; Y4 denotes Y4c or Y4d; where Y4c denotes lower cycloalkyl, optionally substituted with halogen; and Y4d denotes an amino group, optionally substituted with one or more C1-6alkyl; where "aryl" denotes phenyl or naphthyl, "heteroaryl" denotes a monocyclic or bicyclic radical containing 5 to 9 atoms in the ring, which contains at least one aromatic ring containing 5 to 6 atoms in the ring, with one or two N or O heteroatoms, wherein the remaining atoms in the ring are carbon atoms, under the condition that the binding point of the heteroaryl radical is in the aromatic ring, "heterocycloalkyl" denotes a monovalent saturated cyclic radical consisting of one ring containing 5 to 6 atoms in the ring, with one or two ring heteroatoms selected from N, O or SO2. The invention also relates to use of the compound of formula II or a pharmaceutical composition based on the compound of formula II.

EFFECT: obtaining novel compounds that are useful for modulating Btk activity and treating diseases associated with excessive activity of Btk.

7 cl, 2 tbl, 53 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compound of formula I in which R1 represents halogen, methoxy group or cyano group; each of Y1 and Y2 represents CH, and one or two from U, V, W and X represent N, and each remaining one represents CH, or in case X, cam also represent CRa, or Ra represents halogen; A represents CH2CH(OH), CH2CH(NH2), CH(OH)CH(NH2) or CH(NH2)CH2, B represents CH2CH2, CH2NH or CONH, and D represents CH2, or A represents CH(OH)CH2, and B represents CH2NH, N(R2)CO or CONH, and D represents CH2, or B represents N(R2a)CH2, and D represents CH(OH), or A represents CH(OH)CH(OH), B represents CH2NH or CONH and D represents CH2, or A represents CH2CH2, and B represents CH2CH2, CH2NR3, NHCO, CONR4, CH2O, COCH2 or CH2CH2NH, and D represents CH2, or B represents CH2NH, and D represents CO, or A also represents CH2CH2, B represents NR4bCH2 and D represents CH(OH), or A represents CH=CH, B represents CH2NR5 or CONR6, and D represents CH2, or A represents C≡C, B represents CH2NH and D represents CO, or A represents COCH2, B represents CONH and D represents CH2, or A represents CH2N(R7), and B represents CH2CH2, a D represents CH2, or B represents CH2CH(OH), a D represents CH(OH), or A represents NHCH2, and B represents CH2NH, a D represents CH2, or B represents CH2NH, a D represents CO, or A represents NHCO, B represents CH(R8)NH or CH2CH2, and D represents CH2, or A represents OCH2, B represents CH=CH or CONH, and D represents CH2; R2 represents (C1-C4)alkyl; R2a represents hydrogen; R3 represents hydrogen, CO-(CH2)p-COOR3', (CH2)p-COOR3, (C2-C5)acyl or amino(C1-C4)alkyl, or also R3 represents (C1-C4)alkyl, which can be one or two times substituted with hydroxygroup, p stands for integer number from 1 to 4, and R3 represents hydrogen or (C1-C4)alkyl; R4 represents hydrogen or (C1-C4)alkyl; R4b represents hydrogen; R5 represents hydrogen or (C2-C5)acyl; R6 represents hydrogen or (C1-C4)alkyl; R7 represents hydrogen or (C1-C4)alkyl, which can be one or two times substituted with group, independently selected from hydroxygroup and aminogroup, R8 represents hydrogen or (C1-C4)alkyl; E represents one of the following groups (a-a1) where Z represents CH or N, and Q represents O or S, or E represents phenyl group, which is one or two times substituted in meta- and/or para-position with substituents, each of which is independently selected from group, including halogen, (C1-C3)alkyl and trifluoromethyl; or pharmaceutically acceptable salt of such compound. Formula I compound or its pharmaceutically acceptable salt is applied for obtaining medication or pharmaceutical composition for prevention or treatment of bacterial infection.

EFFECT: derivatives of oxazolidine antibiotics for obtaining medication for treatment of bacterial infections.

15 cl, 2 tbl, 214 ex

FIELD: chemistry.

SUBSTANCE: described are 1,2-disubstituted heterocyclic compounds of formula (I) where HET, X, Y and Z values are presented in description, which are phosphodiesterase 10 inhibitors. Also described are pharmaceutical composition and methods of treating central nervous system (CNS) disorders and other disorders, which can influence CNS function.

EFFECT: among disorders that can be subjected to treatment, there are neurological, neurodegenerative and psychiatric disorders, which include, but are not limited by them, disorders, associated with impairment of cognitive ability or schizophrenic symptoms.

14 cl, 824 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula (1) or a salt thereof, where D1 is a single bond, -N(R11)- or -O-, where R11 is a hydrogen atom or C1-C3 alkyl; A1 is C2-C4 alkylene, or any of divalent groups selected from the following formulae , and ,

where n1 equals 0 or 1; n2 equals 2 or 3; n3 equals 1 or 2; R12 and R13 are each independently a hydrogen atom or C1 -C3 alkyl; v is a bond with D1; and w is a bond with D2; D2 is a single bond, C1-C3 alkylene, -C(O)-, S(O)2-, -C(O)-N(R15)-, or -E-C(O)-, where E is C1-C3 alkylene, and R15 is a hydrogen atom; R1 is a hydrogen atom, C1-C6 alkyl, a saturated heterocyclic group which can be substituted with C1-C6 alkyl groups, an aromatic hydrocarbon ring which can be substituted with C1-C3 alkyl groups, C1-C4 alkoxy groups, halogen atoms, cyano groups, a monocyclic aromatic heterocyclic ring containing one or two heteroatoms selected from a group consisting of a nitrogen atom, a sulphur atom and an oxygen atom, or the following formula ,

where n1 equals 0, 1 or 2; m2 equals 1 or 2; D12 is a single bond, -C(O)- or -S(O)2-; R18 and R19 denote a hydrogen atom; R17 is a hydrogen atom or C1-C3 alkyl; and x is a bond with D2; under the condition that when R17 denotes a hydrogen atom, D12 denotes a single bond; under the condition that when D1 denotes a single bond, A1 denotes a divalent group of said formula (1a-5) or (1a-6); when D1 denotes -N(R11)-, -O-, or -S(O)2-, A1 denotes a single bond, C2-C4 alkylene, or any of divalent groups selected from formulae (1a-1)-(1a-3), where, when A1 denotes a single bond, D2 denotes -E-C(O)-; and D3 is a single bond, -N(R21)-, -N(R21)-C(O) - or -S-, where R21 is a hydrogen atom; and R2 denotes a group of formula ,

where Q denotes an aromatic hydrocarbon ring, a monocyclic aromatic heterocyclic ring containing one or two heteroatoms selected from a group consisting of a nitrogen atom, a sulphur atom and an oxygen atom, a condensed polycyclic aromatic ring containing one or two heteroatoms selected from a group consisting of a nitrogen atom, a sulphur atom and an oxygen atom, or a partially unsaturated monocyclic or a condensed bicyclic carbon ring and a heterocyclic ring; and y denotes a bond with D3; and R23, R24 and R25 each independently denotes a hydrogen atom, a halogen atom, a cyano group, C1-C3 alkyl, which can be substituted with hydroxyl groups, halogen atoms or cyano groups, C1-C4 alkoxy group, which can be substituted with halogen atoms, alkylamino group, dialkylamino group, acylamino group, or the formula ,

where D21 denotes a single bond or C1-C3 alkylene; D22 denotes a single bond or -C(O)-; R26 and R27 each independently denotes a hydrogen atom or C1-C3 alkyl; and z denotes a bond with Q; under the condition that when D22 denotes a single bond, R27 is a hydrogen atom. The invention also relates to specific compounds, a pharmaceutical composition based on the compound of formula , a IKKβ inhibitor, a method of inhibiting IKKβ, a method of preventing and/or treating an NF-kB-associated or IKKβ-associated disease, and intermediate compounds of formulae and .

EFFECT: obtaining novel isoquinoline derivatives, having useful biological properties.

46 cl, 3 dwg, 38 tbl, 89 ex

Organic compounds // 2518462

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula

and

,

where X represents S or O, one of X1 and X2 represents CR3' and second represents N or independently CR3', n represents integer number 1, 2 or 3; R1 represents C1-6 halogenalkyl, R2 is selected from halogen and C1-C6-halogenalkyl; R3' represents H, C1-C6-alkyl, halogen, cyanogroup, or phenyl, non-substituted or substituted with halogen, C1-C6-alcoxygroup, C1-C6-halogenalcoxygroup, C1-C6-halogenalkyl group; Z represents halogen, Q radical or group -C(O)-NR5R6; R5 represents H or C1-C4-alkyl, R6 represents H; Q', C1-C6-alkyl, non-substituted or substituted with halogen, cyanogroup, C1-C4-alcoxygroup, C1-C4-alkoxycarbonyl, C2-C4-alkanoyl, aminocarbonyl, N-mono- or N,N-di-C1-C2-alkylaminocarbonyl, C1-C4-alkylthiogroup, group -C(O)NHR7 or radical Q"; or C3-C6-cycloalkyl, substituted with group -C(O)NHR7; or C2-C4-alkinyl; Q, Q' and Q" are such as given in the invention formula; R7 represents C1-C6-alkyl, which is non-substituted or substituted with halogen, cyanogroup, pyridyl; or represents C2-C4-alkinyl. Invention also relates to composition for fighting ectoparasites, containing compound of formula (Ia) or (Ib), and to application of compounds of formula (Ia) or (Ib) for composition production.

EFFECT: compounds of formula (Ia) and (Ib), possessing activity against ectoparasites.

11 cl, 4 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to novel indole and benzomorpholine derivatives of a formula (I) or its pharmaceutically acceptable salt, where R1 represents C1-6-alkyl or C1-3alkyl, substituted with C3-7cycloalkyl; R2 represents halogeno; R3 represents hydrogen; n equals 2, X represents -CH2CH2-O or -CH=CH-; Y represents -O- or -CR4(OH)-; R4 represents hydrogen or C1-3 alkyl. Invention also relates to a pharmaceutical composition based on formula (I) compound and a method of treatment or prevention of the said pathological states.

EFFECT: obtained are novel compounds, which are positive allosteric modulators of matabotropic subtype 2 receptors (mGluR2), which are useful for treatment or prevention of neurological and psychiatric disorders, associated with glutamate dysfunction, and diseases, involving metabotropic subtype 2 receptors GluR2.

22 cl, 2 tbl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula (I), wherein R1 represents an alkoxy group or halogen; each U and V independently represents CH or N; "----" means a bond or is absent; W represents CH or N, or if "----" is absent, then W represents CH2 or NH, provided not all U, V and W represent N; A represents a bond or CH2; R2 represents H, or provided A means CH2, then it also can represent OH; each m and n are independently equal to 0 or 1; D represents CH2 or a bond; G represents a phenyl group that is single or double substituted in meta- and/or para-position(s) by substitutes specified in alkyl, C1-3alkoxy group and halogen, or G represents one of the groups G1 and G2: wherein each Z1, Z2 and Z3 represents CH; and X represents N or CH and Q represents O or S; it should be noted that provided each m and n are equal to 0, then A represents CH2; or a pharmaceutically acceptable salt of such compound. Besides, the invention refers to a pharmaceutical composition for treating a bacterial infection containing an active ingredient presented by a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one therapeutically inert additive.

EFFECT: preparing the oxazolidine compounds applicable for preparing a drug for treating and preventing the bacterial infections.

14 cl, 8 dwg, 2 tbl, 33 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula (I) , where A is a 6-member heteroaryl, having 1 nitrogen atom as a heteroatom, substituted with 2-3 substitutes such as indicated in the claim, R5 is a halogen atom, cyano or C1-C6alkyl, optionally substituted with a halogen atom; R6 is C1-C6 alkyl, optionally substituted with OH; C1-C3 alkenyl; a 5-member heteroaryl, having 2-4 heteroatoms, each independently selected from N, O or S, substituted with 0-2 substitutes such as indicated in the claim, R10 is a 5-member heteroaryl, having 2-3 heteroatoms, each selected from N, O or S, substituted with 0-2 substitutes, which are C1-C3 alkyl; R7, R8, R17 denote a hydrogen or halogen atom. The invention also relates to a pharmaceutical composition, having BK B2 receptor inhibiting activity, which contains compounds of formula (I), a method of inhibiting, a method of localising or detecting the BK B2 receptor in tissue, use of the compounds of compositions to produce a medicinal agent and methods for treatment.

EFFECT: compounds of formula (I) as BK B2 receptor inhibitors.

22 cl, 1 tbl, 54 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a 2,4-diamino-1,3,5-triazine derivative of general formula I, having protein kinase inhibitor properties, use thereof and a pharmaceutical composition based thereon. In general formula I Y is CH2, CHR', O, S, S(O) or S(O)2; X1, X2, X3 are independently selected from a CH groups or N; R1 is a C1-8 aliphatic group, C3-8 cycloalkyl, C6-10 aryl, ethylene-dioxyphenyl, methylene dioxyphenyl, pyridyl, each of which is optimally substituted with one or more identical or different groups R"; R' is hydrogen, OH, halogen, such as F, Cl, Br, I, or carboxyl or carboxamide, optimally N-substituted with (C1-6)alkyl, or cyano or halo(C1-8)alkyl, (C1-8)alkoxy, piperidinyl, optimally substituted with methyl; R" is R' or RD; R21, R22, R23, R24 are independently selected from groups F, Cl, Br, I, CN, (C1-16)alkyl; furthermore, R21 and R22 and/or R23 and R24 can be combined and represent one oxo (=O) group or together with a carbon atom can form a spirocycle containing 3 to 7 carbon atoms; furthermore, R21 and R24 together with two carbon atoms can form an aliphatic or aromatic ring containing 4 to 8 atoms, optionally substituted with one or more groups R'; RD is an oxo group =O or =S.

EFFECT: invention can be used to treat autoimmune or cancerous diseases, rheumatoid arthritis and non-Hodgkin lymphoma.

13 cl, 12 ex

FIELD: biotechnologies.

SUBSTANCE: in a compound of formula ,

X means N or CH, R1 means hydrogen or cyano, R2 means saturated 4-7-membered residue of heterocyclyl, which is bound through a nitrogen atom that contains 1 to 2 heteroatoms chosen from N and O. Besides, heterocyclyl residue can be replaced with one substituent chosen from a group consisting of C3-C6-cycloalkyl, or with 1-4 fluorine atoms. The invention also refers to a method for obtaining compounds and to a medicine on their basis.

EFFECT: compounds can be used for production of a medicine suitable for being used in a method of treatment or prophylaxis of cardiovascular diseases, cardiac insufficiency, anemia, chronic diseases of kidneys and kidney failure.

16 cl, 1 tbl, 29 ex

FIELD: biotechnologies.

SUBSTANCE: invention refers to a compound of formula (I):

,

where R1 represents NR7C(O)R8 or NR9R10; R2 represents hydrogen; R3 represents halogen; R4 represents hydrogen, halogen, cyano, hydroxy, C1-4alkyl, C1-4alkoxy, CF3, OCF3, C1-4alkylthio, S(O)(C1-4alkyl), S(O)2(C1-4alkyl), CO2H or CO2(C1-4alkyl); R5 represents C1-6alkyl (replaced with NR11R12 or heterocyclyl that represents nonaromatic 5-7-membered ring containing 1 or 2 heteroatoms independently chosen from a group containing nitrogen, oxygen or sulphur); R6 represents hydrogen, halogen, hydroxy, C1-4alkoxy, CO2H or C1-6alkyl (possibly replaced with NR15R16 group, morpholinyl or thiomorpholinyl); R7 represents hydrogen; R8 represents C3-6cycloalkyl (possibly replaced with NR24R25 group), phenyl or heteroaryl, which represents aromatic 5- or 6-membered ring containing 1 to 3 heteroatoms independently chosen from the group containing nitrogen, oxygen and sulphur, and which is probably condensed with one 6-membered aromatic or nonaromatic carbocyclic ring or with one 6-membered aromatic heterocyclic ring, where the above 6-membered aromatic heterocyclic ring includes 1 to 3 heteroatoms independently chosen from a group containing nitrogen, oxygen and sulphur; R9 represents hydrogen or C1-6alkyl (possibly replaced with pyrazolyl); R10 represents C1-6alkyl (possibly replaced with phenyl or heteroaryl group, which represents aromatic 5- or 6-membered ring containing 1 or 2 heteroatoms independently chosen from the group containing nitrogen, oxygen or sulphur, and which is possibly condensed with one 6-membered heterocyclic ring, where the above 6-membered aromatic heterocyclic ring contains 1 or 2 heteroatoms independently chosen from the group containing nitrogen, oxygen or sulphur; where the above phenyl and heteroaryl groups in R8, R9 and R10 are possibly independently replaced with the following group: halogen, hydroxy, C(O)R42, C1-6alkyl, C1-6hydroxyalkyl, C1-6halogenoalkyl, C1-6alkoxy(C1-6)alkyl or C3-10cycloalkyl; unless otherwise stated, heterocyclyl is possibly replaced with group of C1-6alkyl, (C1-6alkyl)OH, (C1-6alkyl)C(O)NR51R52 or pyrrolidinyl; R42 represents C1-6alkyl; R12, R15 and R25 independently represent C1-6alkyl (possibly replaced with hydroxy or NR55R56 group); R11, R16, R24, R51, R52, R55 and R56 independently represent hydrogen or C1-6alkyl; or to its pharmaceutically acceptable salts.

EFFECT: new compounds are obtained, which can be used in medicine for treatment of PDE4-mediated disease state.

10 cl, 2 tbl, 202 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to organic chemistry and specifically to 5-phenyl-1H-pyrazin-2-one derivatives of general formula II or pharmaceutically acceptable salts thereof, where R denotes -R1 or - R1-R2-R3; R1 denotes aryl or heteroaryl, and is optionally substituted with one or two R1'; where each R1' independently denotes C1-6alkyl, halogen or C1-6halogenalkyl; R2 denotes -C(=O), -CH2-; R3 denotes R4; where R4 denotes an amino group or heterocycloalkyl, and is optionally substituted with one or two substitutes selected from C1-6alkyl, hydroxy group, oxo group, C1-6hydroxyalkyl, C1-6alkoxy group; Q denotes CH2; Y1 denotes C1-6alkyl; Y2 denotes Y2b; where Y2b denotes C1-6alkyl, optionally substituted with one Y2b'; where Y2b' denotes a hydroxy group, n and m are equal to 0; Y4 denotes Y4c or Y4d; where Y4c denotes lower cycloalkyl, optionally substituted with halogen; and Y4d denotes an amino group, optionally substituted with one or more C1-6alkyl; where "aryl" denotes phenyl or naphthyl, "heteroaryl" denotes a monocyclic or bicyclic radical containing 5 to 9 atoms in the ring, which contains at least one aromatic ring containing 5 to 6 atoms in the ring, with one or two N or O heteroatoms, wherein the remaining atoms in the ring are carbon atoms, under the condition that the binding point of the heteroaryl radical is in the aromatic ring, "heterocycloalkyl" denotes a monovalent saturated cyclic radical consisting of one ring containing 5 to 6 atoms in the ring, with one or two ring heteroatoms selected from N, O or SO2. The invention also relates to use of the compound of formula II or a pharmaceutical composition based on the compound of formula II.

EFFECT: obtaining novel compounds that are useful for modulating Btk activity and treating diseases associated with excessive activity of Btk.

7 cl, 2 tbl, 53 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compound of formula I in which R1 represents halogen, methoxy group or cyano group; each of Y1 and Y2 represents CH, and one or two from U, V, W and X represent N, and each remaining one represents CH, or in case X, cam also represent CRa, or Ra represents halogen; A represents CH2CH(OH), CH2CH(NH2), CH(OH)CH(NH2) or CH(NH2)CH2, B represents CH2CH2, CH2NH or CONH, and D represents CH2, or A represents CH(OH)CH2, and B represents CH2NH, N(R2)CO or CONH, and D represents CH2, or B represents N(R2a)CH2, and D represents CH(OH), or A represents CH(OH)CH(OH), B represents CH2NH or CONH and D represents CH2, or A represents CH2CH2, and B represents CH2CH2, CH2NR3, NHCO, CONR4, CH2O, COCH2 or CH2CH2NH, and D represents CH2, or B represents CH2NH, and D represents CO, or A also represents CH2CH2, B represents NR4bCH2 and D represents CH(OH), or A represents CH=CH, B represents CH2NR5 or CONR6, and D represents CH2, or A represents C≡C, B represents CH2NH and D represents CO, or A represents COCH2, B represents CONH and D represents CH2, or A represents CH2N(R7), and B represents CH2CH2, a D represents CH2, or B represents CH2CH(OH), a D represents CH(OH), or A represents NHCH2, and B represents CH2NH, a D represents CH2, or B represents CH2NH, a D represents CO, or A represents NHCO, B represents CH(R8)NH or CH2CH2, and D represents CH2, or A represents OCH2, B represents CH=CH or CONH, and D represents CH2; R2 represents (C1-C4)alkyl; R2a represents hydrogen; R3 represents hydrogen, CO-(CH2)p-COOR3', (CH2)p-COOR3, (C2-C5)acyl or amino(C1-C4)alkyl, or also R3 represents (C1-C4)alkyl, which can be one or two times substituted with hydroxygroup, p stands for integer number from 1 to 4, and R3 represents hydrogen or (C1-C4)alkyl; R4 represents hydrogen or (C1-C4)alkyl; R4b represents hydrogen; R5 represents hydrogen or (C2-C5)acyl; R6 represents hydrogen or (C1-C4)alkyl; R7 represents hydrogen or (C1-C4)alkyl, which can be one or two times substituted with group, independently selected from hydroxygroup and aminogroup, R8 represents hydrogen or (C1-C4)alkyl; E represents one of the following groups (a-a1) where Z represents CH or N, and Q represents O or S, or E represents phenyl group, which is one or two times substituted in meta- and/or para-position with substituents, each of which is independently selected from group, including halogen, (C1-C3)alkyl and trifluoromethyl; or pharmaceutically acceptable salt of such compound. Formula I compound or its pharmaceutically acceptable salt is applied for obtaining medication or pharmaceutical composition for prevention or treatment of bacterial infection.

EFFECT: derivatives of oxazolidine antibiotics for obtaining medication for treatment of bacterial infections.

15 cl, 2 tbl, 214 ex

FIELD: chemistry.

SUBSTANCE: described are 1,2-disubstituted heterocyclic compounds of formula (I) where HET, X, Y and Z values are presented in description, which are phosphodiesterase 10 inhibitors. Also described are pharmaceutical composition and methods of treating central nervous system (CNS) disorders and other disorders, which can influence CNS function.

EFFECT: among disorders that can be subjected to treatment, there are neurological, neurodegenerative and psychiatric disorders, which include, but are not limited by them, disorders, associated with impairment of cognitive ability or schizophrenic symptoms.

14 cl, 824 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to the field of organic chemistry, namely to compounds of the general formula I , and its pharmaceutically acceptable salts, where R1, R2 and R3 represent hydrogen, D, E, G, J and L represent CH; n equals to an integer number 1 or 2; W represents oxygen; R4 represents C1-6alkyl, C3-6cycloalkyl, C3-6cycloalkenyl, where the said C1-6alkyl is possibly substituted with one substituent, independently selected from a group, consisting of hydrogen, C1-4alkyl, C3-6cycloalkyl and C3-6cycloalkenyl; Y represents carbonyl; R5 represents C1-6alkyl, C1-6alkoxy or C3-4heteroaryl, which represents a heterocyclic aromatic ring, containing 1-2 heteroatoms, selected from nitrogen and oxygen. The invention also relates to a pharmaceutical composition based on a formula I compound, application of the formula I compound and a method of prevention, treatment of alleviation of a disease, associated with abnormal angiogenesis.

EFFECT: obtained are novel compounds useful in treatment of diseases associated with unregulated angiogenesis, such as cancer, as well as skin and eye diseases.

13 cl, 3 tbl, 11 ex

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