Halogenised pyrazolo[1,5-a]pyrimidines (versions), methods for preparing them, based pharmaceutical composition, intermediate compounds and method for preparing them (versions). RU patent 2478101.

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new halogenised pyrazolo[1,5-a]-pyrimidines of general formula (I) and their pharmaceutically acceptable salts possessing affinity with respect to α₁-,α₂ subunits of a GABA_A receptor. In formula R represents alkyl(C₁-C₆); R₁ is specified in a group consisting of alkyl(C₁-C₆) and alkinyl(C₁-C₆); X represents a halogen atom, and Y is specified in a group consisting of -CO- and -SO_2. The invention refers to intermediate enamine compounds and methods for preparing them.

EFFECT: invention also refers to a method for preparing the compounds of formula (I), the based pharmaceutical compounds, to the use of said compounds for preparing said drug preparation for treating or preventing anxiety, epilepsy, sleep disorders, including insomnia, as well as for inducing a sedative-hypnotic effect, anaesthesia and muscular relaxation.

23 cl, 6 tbl, 10 ex

The technical field to which the invention relates

This invention relates to the agents with affinity to the GABA receptor And , in particular to pyrazolo[1,5-a], and more specifically to the [7-(3-amino-4-)-pyrazolo[1,5-a]pyrimidine-3-yl]-thiophen-2-yl- and connections.

The level of technology

Receptor And GABA (gamma-aminobutyric acid (a ) represents a protein that forms a membrane ion channel. GABA receptor And participates in regulation of sedative effect, anxiety, muscle tone, epileptogenic activity and memory functions. These actions are caused by certain of the GABA receptor And , in particular α 1 - and a 2-.

Sedative effect is modulated α 1-subunit. Zolpidem is characterized by a high affinity against α 1-receptors, and its sedative and hypnotic effect is mediated data receptors in vivo. Similarly somnifacient also mediated α-1 receptors.

The anxiolytic effects of diazepam is mediated by strengthening Gamkergicaka transmission in a population of neurons expressing α-2 receptors. This shows that α 2-receptors are targets for treatment of anxiety.

Muscle relaxation using diazepam mainly mediated by the α 2-receptors, as these receptors demonstrate expression in the spinal cord. Protivosudorozhny effect of diazepam is partly due to the α-1 receptors. When using diazepam connection weakening memory, anterograde amnesia is mediated by the α-1 receptors.

A broad overview of the GABA receptor and its α 1 - and a 2-subunits is given in articles .Mohler et al. J. Pharmacol. Exp. Ther., 300, 2-8, 2002); H.Mohler et al. (Curr. Opin. Pharmacol., 1, 22-25, 2001); U. Rudolph et al. (Nature, 401, 796-800, 1999) and D.J.Nutt et al. (Br. J. Psychiatry, 179, 390-396, 2001).

Diazepam and other classic benzodiazepines is widely used as an anxiolytic agents, hypnotics agents, protivosudorozhnykh drugs and muscle relaxants. These side effects include amnesia, reduced physical activity and potentiation of the effects of ethanol.

In this context, connections, in accordance with the present invention, are ligands α 1 - and a 2-of the GABA receptor And intended for clinical use in sleep disorders, preferably with insomnia, anxiety and epilepsy.

Insomnia is a common disease. Its chronic form affects 10% of the population and 30%when taking into account the temporary insomnia. Insomnia characterized by difficulties with falling asleep or sleeping, and is associated with unpleasant effects on the following day, such as fatigue, lack of energy, low concentration and irritability. Action on the social aspects and the state of health of the disease is important and it results in noticeable socio-economic consequences.

Pharmacotherapy in the treatment of insomnia is mainly includes barbiturates and hloralgidrat, but the medicines cause numerous known adverse effects such as toxicity overdose, induction of metabolic and increased dependence and tolerance. They also affect the structure of sleep by reducing the rest of the duration and number of REM sleep stages (stages of REM sleep). Next benzodiazepines have important therapeutic advantage due to their low toxicity, but they still suffer from serious problems of addiction, muscle relaxation, amnesia and reactive insomnia after stopping treatment.

The last known therapeutic approach provides an introduction sleeping pills such as [3,4-b]- (zopiclone), [1,2-a] (zolpidem) and, finally, pyrazolo[1,5-a]pyrimidines (). Later the company started the development of two new pyrazolo[1,5-a] pyrimidines, and , the last significant anksioliticski action. All these connections are faster induction of sleep and have lower unpleasant effects on the following day, over a low potential for addiction and a reduced risk of reactive insomnia than benzodiazepines. The mechanism of action of these compounds is a activation of the GABA receptor And by linking it with the center of the benzodiazepine binding (see article .F.P.George, The Lancet, 358, 1623-1626, 2001). Although benzodiazepines is non-specific ligands center of the GABA receptor binding And , zolpidem and have shown an increased selectivity in respect of α 1-subunit. However, these drugs are still affect the structure of sleep and may induce dependence with prolonged treatment.

The present invention is structurally connected, but has its own patentable relatively connection N-{3-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-, , which is described in the US 6399621, and compounds of N-{3-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl- and N-{3-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]-pyrimidine-7-yl]-phenyl}-N-prop-2-, which are described in the WO 2005014597, examples 3 and 16 respectively, due to their improved characteristics, as shown in section "Detailed description of the invention". Similar connections described earlier in US 4521422.

Study of new active compounds in the treatment of insomnia meet the essential requirements of health, as even the recently introduced hypnotics still affect the structure of sleep and may induce dependence with prolonged use.

It is therefore necessary to focus on developing new hypnotics agents with reduced risk of side effects.

Thus, the present invention is directed at new halogenated pyrazolo[1,5-a] pyrimidines, which are active against GABA And, in particular, in relation to its α 1 - and a 2-subunits. Therefore, the connections matching the given invention is used for treatment and prevention of all these diseases mediated α 1 - and a 2- of the GABA receptor And .

examples of these diseases are sleep disorders, preferably insomnia, anxiety and epilepsy. examples of appropriate indications for connection corresponding to the invention, are all these diseases or conditions, such as insomnia or anesthesia, in which you want to call sleep, cause a sedative effect or cause muscle relaxation.

Disclosure of the invention

In the present invention described a new class of compounds represented by a formula (I):

where R and R 1 , X and Y are defined below, who are of the GABA receptor ligands and their pharmaceutically acceptable salt.

In relation to another object of the present invention is directed to the development of new ways of treating or preventing anxiety, epilepsy and sleep disorders, including insomnia, inducing a sedative-hypnotic effect, anesthesia, sleep and muscle relaxation by the introduction of a therapeutically efficient quantity of the compounds of formula (I) or pharmaceutically acceptable salts. Synthetic methods of obtaining these compounds and a number of intermediate products is also included in the scope of the invention. Themselves corresponding intermediate products are also another object of the invention.

Realization of the invention

The present invention relates to pyrazolo[1,5-a], in particular, to new [7-(3-amino-4-)-pyrazolo[1,5-a]pyrimidine-3-yl]-thiophen-2-yl- and compounds of formula (I):

R represents an alkyl(C 1-C 6 );

R 1 is selected from the group consisting of alkyl(C 1-C 6 )and (C 1-C 6 );

X represents a halogen atom and

Y is selected from the group consisting of CO - and-SO 2 -;

and pharmaceutically acceptable salts.

Preferably, when R means methyl, R 1 is selected from the group consisting of methyl and prop-2-, and X is selected from the group consisting of fluorine and chlorine.

The term "pharmaceutically acceptable salt"used in this context, covers any salt, formed from organic and inorganic acids, such as Hydrobromic, hydrochloric, phosphoric acid, nitric acid, sulfuric acid, acetic acid, adipic, aspartic acid, , benzoic acid, citric acid, , formic acid, fumaric, glutamic acid, lactic, maleic, malic acid, malonic, almond, , 1,5-, oxalic, , propionic, p-, amber, tartaric acid, etc.

The present invention includes compounds:

N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-;

N-{2-chloro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-;

N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-;

N-{2-chloro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl- and

N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-prop-2--.

Another option is the implementation of the present invention is to provide a way of obtaining the compounds of formula (I) and its pharmaceutically acceptable salts.

Connections, in accordance with the present invention may be used for the treatment or prevention of diseases associated with modulation of the GABA receptor And mammals, which is specified in the introduction to the needy in this mammal the effective number of the compounds of formula (I) or pharmaceutically acceptable salts. More specifically, disease associated with modulation of the GABA receptor And include disease associated with modulation of the receptor α 1-GABA-A and/or modulation receptor α 2-And GABA . A non-limiting list of these diseases include anxiety, epilepsy, sleep disorders, including insomnia, etc.

Another option is the implementation of the present invention relates to the application of the compounds of formula (I) for the treatment or prevention of epilepsy in need of this mammal, which includes the introduction of the specified mammal the efficient amount specified connection or pharmaceutically acceptable salts.

Another option is the implementation of the present invention relates to the application of the compounds of formula (I) for the treatment or prevention of violations of sleep in need of this mammal, which includes the introduction of the specified mammal the efficient amount specified connection or pharmaceutically acceptable salts.

Another option is the implementation of the present invention relates to the application of the compounds of formula (I) for the treatment or prevention of insomnia in need of this mammal, which includes the introduction of the specified mammal the efficient amount specified connection or pharmaceutically acceptable salts.

Another option is the implementation of the present invention relates to the application of the compounds of formula (I) to induce a sedative-hypnotic effect of the needy in this mammal, which includes the introduction of the specified mammal the efficient amount specified connection or pharmaceutically acceptable salts.

Another option is the implementation of the present invention relates to the application of the compounds of formula (I) for induction of anesthesia in need of this mammal, which includes the introduction of the specified mammal the efficient amount specified connection or pharmaceutically acceptable salts.

Another option is the implementation of the present invention relates to the application of the compounds of formula (I) to modulate the time required to induce sleep duration and the need of the mammal, which includes the introduction of the specified mammal the efficient amount specified connection or pharmaceutically acceptable salts.

Another option is the implementation of the present invention relates to the application of the compounds of formula (I) to induce muscle relaxation in need of this mammal, which includes the introduction of the specified mammal the efficient amount specified connection or pharmaceutically acceptable salts.

The present invention also relates to a method of treatment or prevention of mammal, suffering from diseases associated with modulation of the GABA receptor And mammals, which consists of the specified needy in this mammal therapeutically efficient quantity of the compounds of formula (I) or pharmaceutically acceptable salt together with a pharmaceutically acceptable diluents, or carriers. In more detail, disease associated with modulation of the GABAA receptor, include disease associated with modulation of the receptor α 1-GABA-A and/or modulated receptor α 2-And GABA . A non-limiting the list of such diseases include anxiety, epilepsy, sleep disorders, including insomnia, etc.

In this context, the term "mammal" will refer to the class Mammalia higher vertebrates. The term "mammal" includes, but is not limited to man.

Another variant of the implementation of the present invention refers to the pharmaceutical compositions comprising a compound of formula (I) or pharmaceutically acceptable salt combined with therapeutically inert media.

Another option is the implementation of the present invention relates to methods of obtaining of intermediate compounds of formula (VI):

where R and R 1 , X and Y such as described above.

Specific intermediate compounds of formula (VI), namely:

N-[5-(3-dimethylamino-)-2-fluoro-phenyl]-N-methyl-;

N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N-methyl-;

N-[5-(3-dimethylamino-)-2-fluoro-phenyl]-N-methyl-methane-sulfonamide;

N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N-methyl-methane-sulfonamide and

N-[5-(3-dimethylamino-)-2-fluoro-phenyl]-N-prop-2--, are another way of implementing the present invention.

Songs include appropriate for oral, rectal, and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most appropriate way will depend on the nature and severity of the condition, which are treated. The most preferred method corresponding to the present invention, is an oral way. The composition can be conveniently presented in unit dosage form is received by any of the methods are well known in the field of pharmacy.

Active connection can be combined with the pharmaceutical carrier in accordance with the approved methods of compiling the pharmaceutical mixtures. Media can take a wide range of forms depending on the form of the preparation required for the application, such as oral or parenteral (including intravenous injection or infusion). In the manufacture of compositions for oral dosage form you can use any of the usual pharmaceutical environments. Conventional pharmaceutical environment include, for example, water, glycols, oils, alcohol, flavouring additives, preservatives, dyes, etc. in the case of oral liquid products (such as, for example, suspensions, solutions, emulsions and elixirs); aerosols or media, such as starch, sugar, microcrystalline cellulose, thinners, agents, sliding substances, binding agents, baking powder and etc. in the case of solid oral drugs (such as, for example, powders, capsules and tablets)and oral solid dosage preferred relative to the oral liquid preparations.

Due to simplicity of application of tablets and capsules are most comfortable single oral dosage form, in this case, use solid pharmaceutical carriers. If necessary, the tablets can be covered by a standard aqueous or non-aqueous methods.

Suitable interval of doses to use is a total daily dose of about 0.01 mg to approximately 100,00 mg, which is taken once per day or, if necessary, in the form of divided doses.

Compounds of General formula (I) can be obtained according to the reaction shown in the Diagram 1.

In intermediate products of formula (II), (R, R 1 , X and Y such, as defined for the formula (I) and Q is a suitable leaving the group, selected from the group consisting of N(dialkyl(C 1-C 6 )), (C 1-C 6 ) and alkoxy(C 1-C 6 ). Preferably, when Q is selected from the group consisting of dimethylamino-, methylthio - and . Processing of the received as a result of compounds in the form of free acid base allows to receive their respective salts.

Reaction (III) with appropriately substituted 1-aryl-2-propen-1-one (II) conduct of inert polar proton or a solvent such as glacial acetic acid, ethanol, methanol, or dimethylformamide dimethyl sulfoxide, if the temperature is in the range from 50 degrees to 130 C. after a few hours (response time) solution is removed, and the remainder is shared between the aqueous solution of sodium bicarbonate and dichloromethane. Raw material, received in the result of the evaporation of organic layer is dry, you can clear one of the following methods: (a) chromatography on silica gel with the use of ethyl acetate or dichloromethane/methanol as eluent or (b) crystallization in a suitable solvent (ethyl acetate, ethanol, methanol, etc).

Intermediate connection formula (II)where Q means [intermediate connection (VI)] to follow the sequence of reactions shown in Diagram 2.

where R and R 1 , X and Y such as described above.

Intermediate compounds of formula (IV)where Y represents the group [intermediate compounds (IV')] receive according to the manner described in article R.H.Uloth et al. J. Med. Chem. 9, 88-96, 1966).

Alkylation of intermediate compounds (IV)leading to intermediate compounds of formula (V), shall be implemented by the education anion and subsequent reaction with .

formula (V) and (VI) receive through the response of relevant (IV) and (V) respectively with N,N- (DMFDMA) or reagent Б (tert-(dimethylamino)methane).

Intermediate compounds of formula (II), where Q is a , Y means and R 1 is a methyl [intermediate compounds (VII)], may alternatively be obtained according to Diagram 3.

The transformation of the compound (IV') (VII) leads to the formation of and at the same time, formation of N- as a result of the use of the properties of N,N- as agent.

Intermediate compounds of formula (II), where Q is a and R 1 is a methyl (X)can be obtained according to Diagram 4.

The advantage of this method is based on the fact that education or sulfonamida carboxamide takes place on the last stage of the method. As a result the total number reaction stages reduce receiving large consignments of products. Moreover, as shown in the diagram, the transformation of compounds (VIII) (IX) leads to the three following reactions in the process held in a single reaction vessel: (a) education ; (b) methylation and (C) with the formation of N-methylated Amin. Subsequent reaction connection (IX) with the corresponding chloride sulfonic acid or carboxylic acid leads to obtaining of intermediate compounds (X).

In accordance with the results obtained compounds presented in the present invention, showed pharmacological activity both in vitro and in vivo, which is similar to or exceeds the activity of the compounds of the prior art. All these results support their use in diseases or conditions, flexible α 1 - and a 2-GABA receptors , such as insomnia or anesthesia, in which you must inducing sleep induction of sedation or the induction of muscle relaxation. Moreover, it was found that when the introduction of compounds relevant to the present invention, in low doses detect the sudden rise sedative- activity relative to that achieved when using compounds, relevant prior art (i.e. and and examples 3 and 16 of the WO 200501497), as demonstrated below.

As will be shown below, installed pharmacological and cytotoxic activity, metabolic stability and pharmacokinetic profile of compounds relevant to the present invention.

a) Pharmacological activity

1-Ligand binding assays. Determination of affinity test compounds for alpha 1 - and a 2-of the GABA receptor And

During the experiment, the use of male mice Sprague-Dawley weighing 200-250 g After remove the cerebellum (fabric, which mainly contains α 1-receptor-GABA (A ) and spinal cord (fabric, which mainly contains α 2-receptor-GABA (A ). Membrane receive according to the method J.Lameh et al. (see Prog. Neuro-Psychopharmacol. Biol. Psychiatry, 24, 979-991, 2000) and .Noguchi et al. (see Eur J Pharm, 434, 21-28, 2002), with minor modifications. After weighing their tissues suspended in 50 mm Tris·HCl (pH 7,4), 1:40 (about./about.) or sucrose 0.32 M in the case of spinal cord, homogenize and then centrifuged at 20000 g for 10 min at 7 degrees C. The resulting in the same conditions and centrifuged again. in conclusion in the minimum volume and stored at -80°C during the night. The next day the process is repeated until the final not in the ratio 1:10 (about./about.) in the case of the cerebellum and the ratio 1:5. (min./about.) in the case of spinal cord.

Affinity determined by competitive tests using flumazenil radioactive labeled as a ligand. Tests carried out according to the methods described in the article S.Arbilla et al. (see Eur. J. Pharmacol., 130, 257-263, 1986) and in article Y.Wu et al. (see Eur. J. Pharmacol., 278, 125-132, 1995) using 96-well plates for . Incubated membrane containing studied receptors flumazenil (with a radioactive label in the final concentration of 1 nm) and the increasing concentration of test compounds (in total 230 mm at 50 mm [pH of 7.4] Tris buffer·HCl). Simultaneously membrane incubated only with radioactive label (total linking 100%) and in the presence of elevated concentrations of flumazenila without a radioactive label (non-specific binding estimate in % of ligand with a radioactive label). Reactions begin by adding a ligand to a radioactive label with the subsequent terms of incubation in 60 minutes at 4 C. at the end of The period of incubation 200 cells reaction is transferred into a tablet for (Millipore) and filtered with a vacuum pump, and then three times washed with cold test buffer. Tablets for are equipped with a filter GF/B, which holds the membrane, containing receptors and ligands with a radioactive label that is associated with the receptor. After washing tablets leave until they are dry. After drying add liquid scintillator and leave it at stirring for the night. The next day tablets count using scintillation counter Perkin-Elmer Microbeta.

For the analysis of the results percentage of specific binding for each concentration test connection count as follows:

% specific binding = (X-N/T-N)x 100

X: the amount of bound ligand for each concentration of the compound.

T: General binding, the maximum number associated with ligand with a radioactive label.

N: the nonspecific binding, number of ligand with a radioactive label associated with nonspecific way, regardless of the receptor.

Each concentration for each connection is tested in three replications and their average values are used to identify the experimental values % specific binding relative to the concentration of the compound. Data on the affinity expressed as % inhibition at concentrations of 10 -5 M and 10 -7 M and receive Ki for a number of compounds, which calculate the ratio between α 1 and C 2 . Results of these tests result in tables 1 and 2. Mainly some connections, in accordance with the present invention, demonstrate higher selectivity as a sedative-hypnotic agents about the activity of muscle relaxation, which is proved by higher ratio α 2 /α 1 compared with the compounds of prior art.

In this context, the ratio of selectivity α 2 /α 1 for connection and example 2 is 9.6 unlike 7,7 for and 5.0 for connection example 3 in WO 2005014597, which thus results in increasing the selectivity 25% and 92%, respectively. Therefore, these compounds expect reduced side effects.

2. In vivo predicted sedative-hypnotics

Effects of these compounds in vivo assessed in the test predicted a sedative-hypnotic effect on mice (see article D.J.Sanger et al., Eur. J. Pharmacol., 313, 35-42, 1996 and G.Griebel et al., Psychopharmacology, 146, 205-213, 1999).

Use the groups of 5-8 male mice CD1 mass 22-26 g at the time of testing. Test connection is administered as a single intraperitoneal doses, suspended in 0.25% of agar with one drop of tween 80 in the volume of 10 ml/kg In every way to test two doses. Control animals are only a carrier. Using the Smart System (Panlab, S.L. Spain) register the distance in centimeters for each click through 5-minute intervals within 30 minutes after intraperitoneal (ip) dosing and 60 minutes after oral (po) dosing. Calculate the percentage reduction of covered distance of treated animals with respect to the control animals (the first 5 minutes of drop). The results of this test are given in table 3.

In vivo sedative- activity in mice.

Connection

% inhibition of motor activity

98 mmol/kg

0,98 mmol/kg

98 mmol/kg

3 mmol/kg

Example 2

Example 4

Example of obtaining 6

Example of getting 8

An example of obtaining 10

for 25.39

Example 3 of WO 2005014597

Example 16 of WO 2005014597

free 71,43

Suddenly, the relevant connections to the present invention show increased sedative-sleeping pill activity than connections, relevant prior art.

In particular, connections, in accordance with the present invention, in low doses cause a sharp increase sedative- activity relative to that achieved when using compounds, relevant prior art (i.e. , and Examples 3 and 16 of the WO 2005014597). This is very important because it is possible to get the desired therapeutic effect (i.e. a sedative-hypnotic) when using a lower dose with the added advantage consists in the fact that can be minimized similar side effects.

Comparison between connections, represented in the present invention, and the respective connections represented in the prior art, shows that the presence of a halogen atom in the structure represented by the formula (I), gives the possibility of increasing the sedative- activity, especially in low doses. For example, when comparing the activity of the compound, the Example 10 of the present invention, with the activity obtained using the compound, the Example 16 of WO 2005014597, reaching an increase of more than 20% when you use low dose regardless of the route of administration.

b) Cytotoxic activity

Definition of in vitro cell toxicity HepG2 within 24 hours

HepG2 cells ( carcinoma person) is obtained from the American type culture collection (ATCC) and cultured in minimum principal environment Igla (Needle) with a balanced salt solution Needle, brought to contain 1,87 mm Glutamax™ I, 0.1 mm amino acids, 1.0 mm sodium pyruvate, 100000 units/l penicillin, 10000 ug/l streptomycin, 90% bovine serum embryos 10%. Water nonradioactive survival analysis cells Promega CellTiter 96 ® includes connecting [3-(4,5--2-yl)-5-(3-)-2-(4-)-2H-, inner salt (MTS). The transformation of MTS in water soluble product performed with the use of enzymes dehydrogenases found in metabolically active cells. The number of product directly proportional to the number of living cells in culture.

Compounds dissolved in DMSO to achieve an initial concentration of 100 mm. Make serial dilution of the stock solution in DMSO to achieve concentrations of 10, 1, 0.1 and 0.01 mm. The mother solution and serial dilution then diluted 1:100 medium for cultivation of cells to get six final concentration for analysis 1000, 100, 10, 1, 0.1, and 0.01 microns. The final concentration of DMSO in all holes is 1%.about. HepG2 cells incubated with the test connections within 24 hours. Relative survival rate determined spectrophotometrically at a wavelength of 490 nm after adding dye MTS and subsequent incubation for one hour. As a positive control use tamoxifen.

The percentage of absorption of samples processed test product, compared with a raw model to calculate the percentage of control. The results of this test lead in table 4.

Accordingly, the compounds obtained in examples 2, 4, 6 and 8, suddenly show more low cytotoxicity as a connection relevant prior art, thus improving the safety profile of compounds relevant to the present invention.

C) Metabolic stability

Definition of in vitro metabolic stability in the cytosolic fraction of human hepatocytes

Compounds dissolved in DMSO, to get the original concentration of 10 mm. Then the mother solution is diluted with solvent and a buffer to obtain the final concentration for analysis of 5 microns. Connection test in the same concentration of 5 microns in two replicas, with 1.0 mg/ml joint person (derived from the company Xenotech ple) at 37 deg C. assess the Metabolism in the presence or absence of cofactors and measured as a reduction of the initial connection through analysis of LC/MS (liquid chromatography/mass spectrometry) at the points 0, 60 and 120 minutes. Then calculate the percentage of the remaining parent compound. Use the General method of the LC:

The mobile phase:

A = 0,1% formic acid in water

In = the 0.1% formic acid in acetonitrile

HPLC column:

Higgins Clipius

With 18 to 5 microns,

50 x 3 mm

Flow rate:

2 ml·min-1

Gradient:

The results of this test are given in table 5.

Metabolic stability in the cytosolic fraction of human hepatocytes

Connection

Example 2

Example 4

Example of obtaining 6

Unexpectedly, a number of compounds relevant to the present invention, have shown an increased metabolic stability compared with the compounds, relevant prior art, predicting, thus, improved pharmacokinetic profile for data connections.

d) the Pharmacokinetic profile

In vivo pharmacokinetic profile after the introduction of single dose

Compound received in example 2, test the pharmacokinetic profile after intravenous administration. used as a reference compound. For each of the connections are three male rats Sprague-Dawley weight 250-300, the Sampling shall be implemented by puncture sinus in the following points in time 2,5, 5, 30, 60, 120, 180, 300 and 420 min after injection. Samples stored in the ice bath to the separation of the plasma. Animals anaesthetize inhalation izoflurana each time the selection of the sample. Plasma is separated by centrifugation (10 min, 4C and 4500 rpm) and stored at a temperature below -70 OC to analysis.

Use analytical method based on the extraction of each connection is by extraction type of fluid-solid with subsequent determination by LC/MS or LC/MS/MS using the internal standard (IS).

Perform account of pharmacokinetic parameters (AUC 0-t = area under the curve from zero to the last point of time of extraction, Cl = clearance, t 1/2 = a half-period of existence and Vd = volume of distribution) according to the analysis without compartmentalisation. The results are presented in table 6.

Pharmacokinetic parameters

Parameter

Connection

Example 2

of 0.0011

Example 2

t 1/2 (h)

Example 2

Vd (l/kg)

Example 2

Experimental results show a completely different pharmacokinetic profile for the connection, presented in example 2, compared with a connection, the relevant prior art - . Indeed, the area under the curve 57% of connections, presented in example 2, indicating an increased level of impact of a product; clearance is reduced by 20%, while its half-life existence increased by 76%, thus showing slow speed out, and the final volume of distribution above 212%, suggesting widespread in deep nonaqueous departments (i.e. the brain) compared with . Pharmacokinetic parameters correlate with the number of facts found in pharmacology animals. For example, in a test in vivo sedative- activity in mice (3 mmol/kg) percentage inhibition reduced from 74% (5 minutes) to 67% (60 minutes) for , on the contrary, the specified parameter remains constant at the level of 84% for the connection of the obtained in example 2. These unexpected pharmacokinetic properties show that the link corresponding to the present invention provides improved quality of sleep, avoiding in this way, night awakenings, and ensuring a healthy and prolonged sleep.

The following examples illustrate the volume of the present invention.

Example 1: N-[5-(3-dimethylamino-)-2-fluorophenyl]-N-

3.3 g (16.9 per mmol) N-(5-acetyl-2-fluorophenyl)- dissolved in at 8.36 ml (7,49 g) (62,89 mmol) N,N- and the resulting solution is heated under reflux for 6.5 hours. Excess flying reagent is removed by distillation under reduced pressure, receiving raw material, which is crystallized of ethyl acetate. Get 3,32 g N-[5-(3-dimethylamino-)-2-fluoro-phenyl]- as a solid, yellowish-white in color (output 78,6%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 2,21 (3H, s), 2,89 (3H, s), 3,11 (3H, s), 5,65 (1H, d, J=12,8 Hz), 7,05-7,1 (1H, m), a 7.62-7,68 (2H, m), to 7.77 (1H, d, J=12,4 Hz), 8,71-8,73 (1H, m).

MC (ES) m/z=251 (MH+).

HPLC=99,8%.

1.5 g (5,99 mmol) N-[5-(3-dimethylamino-)-2-fluorophenyl]- dissolved in 15 ml of dry N,N-dimethylformamide. , Which is formed solution at 0 degrees C in an inert atmosphere add 0.29 g (7,31 mmol) hydride sodium. After stirring for 30 minutes, add the solution 0,94 g (6,59 mmol) iodide methyl 5 ml dry N,N-dimethylformamide and continue mixing at room temperature for 5 hours. The solvent is removed by distillation under reduced pressure. The resulting residue add 30 ml of dichloromethane and 10 ml of water. Separate the two layer and the water layer is washed with 30 ml dichloromethane. Organic layers washed with 40 ml of water and dried over magnesium sulfate. Layer dichloromethane is evaporated to dryness, getting the oil, which at the crystallization of ethyl acetate gives 804 mg N-[5-(3-dimethylamino-)-2-fluoro-phenyl]-N-methyl- in the form of yellowish-white solid (output 50,8%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 1,85 (3H, s), 2,94 (3H, s), the 3.17 (3H, s), 3,22 (3H, s), 5,62 (1H, d, J=12,4 Hz), 7,16-7,25 (1H, m), 7,78-7,89 (3H m).

MC (ES) m/z=265 (MH+).

HPLC=94,9%.

Example 2: N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-

Mixture 0,073 g (0.38 mmol) (5-amino-1H-pyrazol-4-yl)thiophene-2-Il- and 0.1 g (0.38 mmol) N-[5-(3-dimethylamino-)-2-fluorophenyl]-N- in 10 ml of glacial acetic acid is heated under reflux for 2.5 hours and then the solvent is removed by distillation under reduced pressure. The resulting residue add 15 ml of dichloromethane and 10 ml saturated sodium bicarbonate solution. Two layers are separated and the water layer washed with 10 ml dichloromethane. Organic layers washed with 10 ml of water and dried over magnesium sulfate. Layer dichloromethane evaporated to dryness, getting the oil, which in the presence of ethyl acetate gives 112 mg N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl- as a solid (exit 75%).

Data 1 H NMR spectroscopy(400 MHz, CDCl 3 ): δ 1,98 (3H, s,), 3.3V (3H, s), 7,13 (1H, d, J=4 Hz), 7,18-7,20 (1H, m), 7,42 (1H, t, J=8,8 Hz), 7,71 (1H, d, J=5,2 Hz), 8,02-8,08 (2N, m), 8,12 (1H, dd, J=2.4 and 7.6 Hz), 8,71 (1H, s), 8,82 (1H, d, J=4 Hz).

MC (ES) m/z=395 (MH+).

HPLC=99,2%

Etc. (melting) = 165-167°C

Example 3: N-[2-Chloro-5-(3-dimethylamino-)-phenyl]-N-

4,46 g (21.1 mmol) N-(5-acetyl-2-chlorophenyl)- dissolve 10.4 ml (9,34 g) (78,39 mmol) N,N- and the resulting solution is heated under reflux for 6.5 hours. Excess flying reagent is removed by distillation under reduced pressure, receiving raw material, which is crystallized of ethyl acetate. Get 4,53 g N-[2-chloro-5-(3-dimethylamino-)-phenyl]- as a solid, yellowish-white in color (output 80,5%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 2,24 (3H, s), 2,90 (3H, s), 3,12 (3H, s), 5,66 (1H, d, J=12,4 Hz), 7,38 (1H, d, J=8,8 Hz), 7.62 mm (1H, d, J=8,8 Hz), 7,69 (1H, s), 7,77 (1H, d, J=12,4 Hz), 8,7 (1H, s).

MC (ES) m/z=267 (MH+).

HPLC=98,3%.

1,0 g of (3.75 mmol) N-[2-chloro-5-(3-dimethylamino-)-phenyl]- dissolved in 10 ml of dry N,N-dimethylformamide. To an educated solution at 0 degrees C in an inert atmosphere add 0.18 g (4,57 mmol) hydride sodium. After stirring for 30 minutes, add the solution 0.59 g (4,12 mmol) iodide methyl 3 ml of dry N,N-dimethylformamide and continue mixing at room temperature for 5 hours. The solvent is removed by distillation under reduced pressure. The resulting residue add 30 ml of dichloromethane and 10 ml of water. Separate the two layer and the water layer is washed with 30 ml dichloromethane. Organic layers washed with 40 ml of water and dried over magnesium sulfate. Layer dichloromethane is evaporated to dryness, getting the oil, crystallization of ethyl acetate-hexane receive 928 mg N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N- as a solid, yellowish-white in color (output 88,16%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 1,79 (3H, s), 2,94 (3H, s), the 3.17 (3H, s), 3,19 (3H, s), 5,61 (1H, d, J=12,4 Hz), 7,50 (1H, d, J=8,4 Hz), 7,79 is 7.85 (3H m).

MC (ES) m/z=281 (MH+).

HPLC=100%.

Example 4: N-{2-Chloro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-

1.66 g (6,77 mmol) N-(5-acetyl-2-fluorophenyl)-N--sulfonamida dissolved in 3,35 ml (3.0 g) (25,18 mmol) N,N- and the resulting solution is heated under reflux for 2.5 hours. Mixture to cool to room temperature. To an educated solid substance add 20 ml of n-hexane and filtered, receiving solid, which is crystallized of ethyl acetate. Get 1,37 g N-[5-(3-dimethylamino-)-2-fluorophenyl]-N- as a solid, yellowish-white in color (output 67,4%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ of 2.92 (3H, s), 2,96 (3H, s)and 3.15 (3H, s), 3,31 (3H, s), 5,61 (1H, d, J=12,8 Hz), 7,13-7,18 (1H, m), 7,78 (1H, d, J=12,8 Hz), of 7.88-7,93 (2N, m).

MS (ES) m/z=301 (MH+).

HPLC=97,99%.

Example of obtaining 6: N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-

Mixture 0,064 g (0.33 mmol) (5-amino-1H-pyrazol-4-yl)thiophene-2-Il- and 0.1 g (0.33 mmol) N-[5-(3-dimethylamino-)-2-fluorophenyl]-N- in 10 ml of glacial acetic acid is heated under reflux for 2.5 hours and then the solvent is removed by distillation under reduced pressure. The resulting residue add 15 ml of dichloromethane and 10 ml saturated sodium bicarbonate solution. Separate the two layer and the water layer is washed with 10 ml dichloromethane. Organic layers washed with 10 ml of water and dried over magnesium sulfate. Layer dichloromethane is evaporated to dryness, getting the oil, which in the presence of ethyl acetate gives 111 mg N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl- Amida as a solid (exit 77%).

Data 1 H NMR spectroscopy(400 MHz, CDCl 3 ): δ 3,01 (3H, s,), 3,39 (3H, s), 7,13 (1H, d, J=4,4 Hz), 7,18-7,20 (1H, m), 7,36 with 7.41 (1H, m), 7,70 (1H, dd, J=1,2 and 5.2 Hz), 8,07-8,09 (1H, m), 8,11-8,17 (2N, m), 8,7 (1H, s), 8,80 (1H, d, J=4,8 Hz).

MC (ES) m/z=431 (MH+).

HPLC=98,6%.

Etc.=194-196°C

Example of getting 7: N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N-

1.0 g (4,04 mmol) N-(5-acetyl-2-chloro-phenyl)- dissolved in 10 ml of dry N,N-dimethylformamide and 2,69 ml (2,41 g) (20,19 mmol) N,N-dimethylformamide . The resulting solution is heated under reflux for 2 hours. Solvent and excess flying reagent is removed by distillation under reduced pressure, getting the oil, which in the presence of ethyl acetate gives 1,04 g of raw cotton. It was chromatographed (silica gel), using ethyl acetate/2-propyl alcohol as a solvent. Get 0.51 g

N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N-methyl- as a solid yellowish-white (exit 40%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 2,9 (3H, s), 3,04 (3H, s)and 3.15 (3H, s), 3.3V (3H, s), 5,61 (1H, d, J=12,4 Hz), of 7.48 (1H, d, J=8,4 Hz), 7,78 (1H, d, J=12,8 Hz), 7,83 (1H, dd, J=8,8-1.6 Hz), to 7.93 (1H, d, J=1.6 Hz).

MC (ES) m/z=317 (MH+).

HPLC=87,58%.

Example of getting 8: N-{2-chloro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-

Mixture 0,076 g (0,39 mmol) (5-amino-1H-pyrazol-4-yl)thiophene-2-Il- and 0.124 g (0,39 mmol) (N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N- in 10 ml of glacial acetic acid is heated under reflux for 1.5 hours and then the solvent is removed by distillation under reduced pressure. The resulting residue add 15 ml of dichloromethane and 10 ml saturated sodium bicarbonate solution. Separate the two layer and the water layer is washed with 10 ml dichloromethane. Organic layers washed with 10 ml of water and dried over magnesium sulfate. Layer dichloromethane is evaporated to dryness, getting the oil, which in the presence of ethyl acetate gives 128 mg

N-{2-chloro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl- as a solid (exit 73%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 3,09 (3H, s,), 3,38 (3H, s), 7,15 (1H, d, J=4,8 Hz), 7,19-7,20 (1H, m), 7,68-7,71 (2H, m), 8,07-8,09 (2H, m), 8,19 (1H, d, J=2 Hz), 8,71 (1H, s), 8,82 (1H, d, J=4,4 Hz).

MC (ES) m/z=447 (MH+).

HPLC=98,1%.

Etc.=241 to 243°C.

An example of obtaining 9: N-[5-(3-dimethylamino-)-2-fluorophenyl]-N-prop-2-

1.2 g (4,46 mmol) N-(5-acetyl-2-fluorophenyl)-N-prop-2-- dissolve in 3 ml (2.7 g) (22,58 mmol) N,N- and heated under reflux for 2.5 hours. The mixture is cooled at room temperature and add 20 ml of n-hexane. The oil obtained was chromatographed (silica gel), using ethyl acetate/2-propyl alcohol as a solvent. Get 0,46 g of the solid substance, yellowish-white in color. This solid crystallized in ethyl acetate and receive 0,213 g N-[5-(3-dimethylamino-)-2-fluoro-phenyl]-N-prop-2-- (output 14,7%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 2,35 (1H, m), 2,92 (3H, s), 3,11 (3H, s)and 3.15 (3H, s), 4,43 (2H, m), 5,61 (1H, d, J=12,8 Hz), 7,16-7,21 (1H, m), 7,79 (1H, d, J=12,8 Hz), one-7.91-7,94 (1H, m), 8,01-8,04 (1H, m).

MC (ES) m/z=325 (MH+).

HPLC=for 91.63%.

An example of obtaining 10: N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-prop-2--

N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-prop-2-- as a solid (exit 61%).

Data 1 H NMR spectroscopy (400 MHz, CDCl 3 ): δ 2,39 (1H, s)at 3.16 (3H, s,), 4,50 (2H, s), 7,14 (1H, d, J=4,4 Hz), 7,18-7,20 (1H, m), 7,40-7,44 (1H, m), 7,70 (1H, m), 8,07-8,09 (1H, m), 8,18-8,21 (1H, m), 8,24-of 8.26 (1H, m)of 8.7 (1H, s), 8,80 (1H, d, J=4,8 Hz).

MC (ES) m/z=455 (MH+).

HPLC=94,9%.

Etc.=149-153°C.

An example of a composition of 1: 5 mg Tablets

Link corresponding example 2

Colloidal dioxide silicon

Croscarmellose sodium

Magnesium stearate

Polysorbate 80

Hydroxypropylmethylcellulose

Polyethylene glycol 4000

Titanium dioxide E171

Microcrystalline cellulose q.s.* to

* as needed

Example 2 composition: Capsules 10 mg

Link corresponding example 2

Colloidal silicon dioxide

Crospovidone

Magnesium stearate

Sodium lauryl sulfate

Titanium dioxide E171

Indigotin 132

Microcrystalline cellulose q.s. to

An example of songs 3: Oral drops

Link corresponding example 2

Propylene glycol

Sodium saccharin

Polysorbate 80

Lemon flavoring

Purified water q.s. to

An example of songs 4: Tablets 2.5 mg

Link corresponding example 2

Colloidal silicon dioxide

Croscarmellose sodium

Magnesium stearate

Polysorbate 80

Hydroxypropylmethylcellulose

Polyethylene glycol 4000

Titanium dioxide E171

Microcrystalline cellulose q.s.* to

An example of songs 5: 5 mg Capsules

Link corresponding example 2

Colloidal silicon dioxide

Crospovidone

Magnesium stearate

Sodium lauryl sulfate

Titanium dioxide E171

Indigotin 132

Microcrystalline cellulose q.s. to

An example of songs 6: Oral drops

Link corresponding example 2

Propylene glycol

Sodium saccharin

Polysorbate 80

Lemon flavoring

Purified water q.s. to

1. Halogenated pyrazolo[1,5-a]pyrimidines, covered by the General structural formula (I)

where R represents an alkyl(C 1-C 6 ); R 1 is selected from the group consisting of alkyl(C 1-C 6 ) and (C 1-C 6 ); X represents a halogen atom and Y is selected from the group consisting of-SO - and-SO 2 -, and pharmaceutically acceptable salt.

2. Pyrazolo[1,5-a]pyrimidines of claim 1, wherein R means methyl.

3. Pyrazolo[1,5-a]pyrimidines of claim 1, wherein R 1 means methyl.

4. Pyrazolo[1,5-a]pyrimidines of claim 1, wherein R 1 means prop-2-.

5. Pyrazolo[1,5-a]pyrimidines according to claim 1, where X stands for fluorine.

6. Pyrazolo[1,5-a]pyrimidines according to claim 1, where X stands for chlorine.

7. Halogenated pyrazolo[1,5-a]pyrimidines, selected from the group consisting of: N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-; N-{2-chloro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-; N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl-; N-{2-chloro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-methyl- and N-{2-fluoro-5-[3-(thiophen-2-carbonyl)-pyrazolo[1,5-a]pyrimidine-7-yl]-phenyl}-N-prop-2--; and its pharmaceutically acceptable salt.

8. The method of obtaining pyrazolo[1,5-a]pyrimidines of General formula (I) according to claim 1, in which the interaction of intermediate compound of formula (II)

where R and R 1 , X and Y such, as defined for the formula (I), a Q appropriate means leaving the group, selected from the group consisting of di( 1-C 6 )amino-group, (C 1-C 6 )group and alkoxy(C 1-C 6 )group, with intermediate compound of formula (III)

and alternatively interact generated in the reactions of the compounds in the form of free base with acid to obtain the salts of these compounds.

9. The method according to item 8, in which the use of intermediate compounds of formula (II), where Q is selected from the group consisting of , and .

10. Pharmaceutical composition, possessing affinity against α 1 -, a 2 subunits of the GABA receptor , including a therapeutically effective amount of pyrazolo[1,5-a]pyrimidines according to claim 1, together with adequate quantities of pharmaceutical fillers or media.

11. Application of pyrazolo[1,5-a]pyrimidines according to claim 1 for the receipt of a medicament for the treatment or prevention of anxiety at a mammal.

12. Application of pyrazolo[1,5-a]pyrimidines according to claim 1 for the receipt of a medicament for the treatment or prevention of epilepsy at a mammal.

13. Application of pyrazolo[1,5-a]pyrimidines according to claim 1 for the receipt of a medicament for the treatment or prevention of violations of sleep at a mammal.

15. Application of pyrazolo[1,5-a]pyrimidines according to claim 1 for obtaining of medicinal product to induce a sedative-hypnotic effect at a mammal.

16. Application of pyrazolo[1,5-a]pyrimidines according to claim 1 for obtaining of medicinal products for anesthesia at a mammal.

17. Application of pyrazolo[1,5-a]pyrimidines according to claim 1 for receiving the drug for the modulation of the time needed to induce sleep and its duration at a mammal.

18. Application of pyrazolo[1,5-a]pyrimidines according to claim 1 for obtaining of medicinal product to induce muscle relaxation at a mammal.

19. Intermediate compounds selected from the group consisting of: N-[5-(3-dimethylamino-)-2-fluorophenyl]-N-; N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N-; N-[5-(3-dimethylamino-)-2-fluorophenyl]-N-methyl-; N-[2-chloro-5-(3-dimethylamino-)-phenyl]-N--sulfonamide and N-[5-(3-dimethylamino-)-2-fluorophenyl]-N-prop-2--.

20. The method of obtaining intermediate compound of formula (VI)

where R and R 1 , X and Y such, as defined above, in which the interaction of the acetophenone formula (IV)

where R, X and Y such, as defined above, with N,N-dimethylformamide- or tert-(dimethylamino)methane, and then spend alkylation obtained during the above reaction formula (V')

where R, X and Y such, as defined above, through education anion with hydride compound, such as NaH, then carry out interaction of the resulting reaction products with formula ZR 1 , where Z means halogen atom, and R 1 means alkyl(C 1-C 6 ).

21. The method according to claim 20 in which hydride connection represents the sodium hydride, and Z mean iodine.

22. The method of obtaining intermediate compound of formula (VI), in which the interaction of the acetophenone formula (V)

where R and R 1 , X and Y such, as defined above, with N,N-dimethylformamide- or tert-(dimethylamino)methane.

23. The method of obtaining intermediate compound of formula (VII)

where R and X such, as defined above, in which the interaction acetophenone formula (IV')

with N,N-dimethylformamide-.