Derivatives of dipeptides or their pharmaceutically acceptable salts, methods for their production, antihyperlipidemia pharmaceutical composition

 

(57) Abstract:

Usage: medicine for the treatment of hypertension. The essence of this invention are derivatives of dipeptides of General formula I:

,

including tautomeric forms, where n = 0 or 1; R = halogen, OH, SH, SR4OR4where R4is lower alkyl or phenyl, with other5-, where R5is phenyl or acetyl, di/lower alkyl/ amino, COOH or COO /lower/ alkyl; R1= OH; /low/ alkoxyl, R2= lower alkyl, R3= halogen, NO2, (lower) alkyl, aryl /lowest/ the alkyl or their pharmaceutically acceptable salts, 2 variants of the method of their derivation; A/ by reacting the compound (II):

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with the dipeptide of formula III:

< / BR>
in the presence of N,N-dicyclohexylcarbodiimide followed, if necessary, conversion of compound I, where R = Cl, compound I, where R = SH and, if necessary, by hydrolysis, by salt formation of esters or diesters obtained in this manner; B/ by reacting compounds II':

,

where X = halogen, the rest of the values specified above, except R = SH, with the dipeptide III, the process is conducted in the presence of organic bases, if R1= OH, in the presence of inorganic bases and, if necessary, with subsequent conversion of compounds I, the slot data; and - antihyperlipidemia pharmaceutical composition comprising as an active component I in an effective amount. 4 C. and 6 C.p. f-crystals, 6 PL.

The invention relates to new N-substituted pyridinyl)caronel-dipeptides, which have activity as inhibitors of the enzyme that promotes the conversion of angiotensin. These new derivatives can come along with suitable for pharmaceutical use by the media in the form of pharmaceutical compositions that can be used in the treatment of hypertension and other cardiovascular diseases, pathophysiology which includes a system the renin-angiotensin-aldosterone.

In the 70-ies in the pharmaceutical treatment of high blood pressure has been significant progress related to development tools, has a direct impact on the system the renin-angiotensin and kallikrein-kinin, and especially after synthesis of the first compounds, effectively ingibirovali enzymatic conversion of Decapeptide of angiotensin I into a powerful vasopressor tool (narrowing of blood vessels) angiotensin II, i.e, compounds acting as inhibitors of the enzyme that promotes the conversion of angiotensin (ACE, Peptide of bradykinin. In experiments on animals and man discovered that some inhibitors of the FCA can inhibit the vasoconstrictor effects of injected angiotensin I and have activity against high pressure when tested in animal models and patients suffering from hypertension. Demonstrated their suitability for the treatment of chronic heart disease.

In U.S. patent [1] described N-ACI-derivatives of alpha-amino acids, which are effective inhibitors of the FCA, which as such may be useful for the treatment of hypertension. More specifically, these compounds are mercapto-derivatives of N-acyl-L-Proline, including as the most representative member of this series, for example, D-3-mercapto-2-methylpropanoyl-L-Proline or captopril, the first protivogipertonicheskoe tool, based on the inhibition of the FCA and is suitable for oral administration, which became internationally popular drug.

The second important success in this area, although based on completely different approach is to develop compounds described in European patent application [2] which are carboxyethyl-dipeptide derivatives, the most characteristic is tours of the FCA and the corresponding significant structural features of the potential inhibitors, and also of interest to obtain new compounds with different activity, kinetics and/or toxicity, has led to the development of new classes of inhibitors of the FCA. From a number of studies of the relationship between activity and structure it was concluded that effective inhibition of the enzyme can be achieved only in the case of a molecule that contains at least three distinctly characteristic of an area or areas that meet the following General structure:

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Area typically contains A carboxyl group in the alpha position, which is tightly associated with the cationic area structure of the enzyme. Several studies have found that L-Proline is the optimal structure for this area or part, although it is also possible presence of his pyrolidine ring in a modified form.

Zone B must contain the functional group with specific activity against the binding of the cation Zn++ located in the "active zone" of the enzyme. Group binding of zinc, usually acidic in nature, can be a mercapto group (as in the case of captopril and its analogues) or carboxyl group (as in the case of enalaprilat, lisinopril and their analogues), as well as any group-the end of the FCA with groups predecessors include allti-derivative alacepril, pivaloyl and carboxylic esters enalapril and perindopril. Some classes of inhibitors contain other acidic groups in the area B, such as-P(O)(OH)- or-P(O)(OH)-O - as in free form and in the form of esters. In any case, all known ligands for zinc necessarily associated with an alkyl group or, in some cases with cycloalkyl group, but are never part of an aromatic structure (M. J. Vibrat, A. A. Patchett, "Medicinal Research Reviews", 6, 483-531, 1985).

Area C plays a role of a bridge between the active areas of zones A and B and undoubtedly must meet specific stereochemical requirements, since the most active compounds contain the element that is derived from L-amino acids (such as L-alanine or L-lysine) in the corresponding dipeptide structure. In the group type of captopril similar stereochemical structure may exist if the atom C-2 2-methylpropanoyl link.

Such a simplified General model applicable to almost all the different structures of known inhibitors of the FCA, although the special or additional requirements must be met for each specific chemical options. In this context, the most well-known correlation between structure and activity series mercapto-acyl-and what each of the three structural parts of the implemented system changes it should be noted that for both of these General series of different attempts to find a binding zinc-groups other than mercapto - or carboxyl groups, has resulted in generally inactive compounds or at least to a considerable loss of activity of the obtained compounds. In particular, in the class of dipeptide derivatives are effective as inhibitors of the FCA, the use of N-substituted carboxamido or thioamides as ligands for zinc according to the prior art is not provided.

The present invention provides a new dipeptide derivatives with activity against inhibition of the FCA and having as main and distinguishing characteristics of the structure, consisting of a pyridine ring, which is linked to a functional group having the potential to bind zinc, specifically as alpha-substituent, such as an OH group, SH3, NH2and COOH or functionally related group or group-predecessor. Such alpha-substituted pyridine structure, connected through the carboxyl group to the terminal amino group of the dipeptide, is located in part of the overall structure, the relevant discussion is in the junction zone C.

Due to these specific structural differences dipeptide derivatives may not be included and not included under one of the General classes or any specific chemical series described to date, according to the prior art as inhibitors of the FCA.

Within the General class of dipeptide compounds have a distinct difference from carboxialkilnuyu of dipeptides, since the latter contain arylcarboxamide structure. Specified structural grouping is very unusual for the field of natural or synthetic inhibitors of the FCA. In addition,(alpha-substituted pyridinyl) carbonyl derivatives of dipeptides is not described and not even assumed in the literature according to the available prior art that can be set, for example, from classic reviews in this area, composed of E. C. of Petrella and M. A. Ondetti ("Medicinal Research Reviews", 2, 1-41, 1982) and the above-noted work of M. J. Vibrate and A. A. of Patchett.

Further, from a practical point of view should take into account that carboxialkilnuyu dipeptide inhibitors according to the prior art contain asymmetric carbon atom (in zone B) in addition to the two asimmetricheskii outputs due to the necessary separation of the optical isomers of the crude products. Since the compounds according to the present invention do not have such additional asymmetric carbon atoms, they can be easily obtained with good outputs by introducing pyridinoline carbonyl of residue in the target dipeptide through his terminal NH group.

In its broadest aspect the present invention relates to new derivatives dipeptides General formula (I):

< / BR>
including tautomeric forms,

where n represents 0 or 1;

R represents OH, SH, halogen, OR4, SR4;

where R4is lower alkyl or phenyl, with other5where R5- phenyl, acetyl, and (lower alkyl)amino; COOH, COO(lower)alkyl;

R1represents OH, (lower)alkoxy,

R2lower alkyl;

R3halogen, NO2, (lower)alkyl, aryl-(lower)alkyl or their pharmaceutically acceptable salts.

R3in the formula (I) can take any free position in the pyridine ring.

Similarly substituted pyridine ring may be associated with carbonyl-dipeptide of the substructure through the alpha-, beta - or gamma-position of the pyridine ring, one of the alpha-positions which is already occupied by a group R.

In number when 1 represents OH and/or R represents COOH, salts of alkali metals such as sodium or potassium, or salts of alkaline-earth metals such as calcium. This includes also ammonium salts, for example salts with hydroxide ammonium, substituted amines or basic amino acids.

As is well known to experts in the field of nitrogen-containing heterocycles, derivatives of pyridine, alpha-substituted by hydroxyl or mercapto groups, usually exist in tautomeric forms, i.e., in the form of 2-pyridinone and 2-pyridines, respectively, i.e., in the form of special types of cyclic carboxamides and thioamides. From this point of view it should be understood that compounds of General formula (I) in which R represents OH or SH, can also be tautomerism in which the corresponding amide forms are usually basic.

Compounds of General formula (I) are inhibitors of the enzyme that promotes the conversion of angiotensin, and can be used as a means of lowering the pressure (protivogipertonicheskoe funds) in the treatment of mammals, including humans. They can also be used in the treatment of chronic heart disease and other diseases that have pathophysiologically pharmaceutical composition, containing at least one compound of General formula (I) in combination with one or more suitable for pharmaceutical use carriers or excipients and may optionally additives and/or additional components, etc. in liquid or solid form, and preferably in the form of unit dosages. Compositions according to the present invention it is most convenient to use the oral method of administration, although there are also other ways, such as parenteral, rectal or introduction through inhalation, which in some cases can be even more preferable. The pharmaceutical compositions according to the present invention can be obtained in accordance with customary methods, for example, by simple mixing of the components of the desired composition. In a number of suitable carriers or excipients are the usual agents of this kind are well known to specialists in this field. Pills introduction most preferably is carried out using the compositions in the form of tablets (coated or uncoated), capsules, or liquids such as solutions, syrups or suspensions. Solid formulations for oral apollonivna steps.

For the treatment of hypertension and/or possibly other diseases for which treatment may be useful biological activity of these new compounds according to the present invention, the dosage level is in the range from 2 to 1000 mg/day to the patient in the form of single or fractional doses, although the individual dose for each patient depends on the activity of the specific compound, the type and degree of development of the disease, as well as such individual patient, as the body weight and gender, as well as from other factors that are commonly understood by a person skilled in this field. In the treatment of hypertension dose preferably ranges from 5 to 500 mg/day per patient.

Compounds according to the present invention can also be used in combination with other active pharmaceutical point of view connections, for example with protivogipertonicheskoe means or other means, used in cardiovascular therapy, for example with diuretics or beta-adrenergic blocking agents. These other active compounds can also be included in the pharmaceutical compositions according to the present invention together with the new connection a few from among the following methods. For the person skilled in the art it is obvious that you can adapt for this purpose, and also other synthetic methods well known in the field of peptide chemistry.

The first method (method A) includes the reaction of a combination of carboxylic acids of General formula (IIa) with a dipeptide of the General formula (III):

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in which R, R1R2and n have the same meaning as before, but with the exception of COOH in the case of R and OH in the case of R1in the presence of preferably equimolar amount of a suitable agent combinations, such as a carbodiimide and particularly preferably N,N'-dicyclohexylcarbodiimide (DCC). Usually the reaction is carried out in a suitable organic solvent of the basic character at room temperature.

Dipeptide of the General formula (III) preferably used in the form of ester, for example ester derived lower alkyl. The free amine group can be optionally transferred to salt, for example,by means of hydrochloric acid. Examples of suitable basic organic solvents include pyridine and a mixture of tertiary aliphatic amines (e.g. triethylamine) in an inert, preferably halogen-substituted solvent (for example, chloroform the imposition of the General formula (I) in the form of monoether (L-prolinnova substructure) in the case when R represents a group other than COOR4, or in the form of diapir, when used as a compound of General formula (IIa), for which R= COOR4, (R4 alkyl, aryl).

You should understand that in the case when the source dipeptide has an additional amino group, i.e., when R2 is an amino(lower alkyl) specified additional amino group must be present in protected form, for example, it can be protected by benzyloxycarbonyl group or any other group that can be easily removed later according to standard techniques known to the expert in this field.

According to a variant of the method A can be used almost the same technique, but with replacement of the dipeptide of the General formula (III) in the esterified amino acid of General formula HN-CH(R2)-COR1. Thus obtained ester N-(alpha-substituted pyridinyl)-carboalumination can hydrolyze and enter the resulting free amino acid in the reaction with a complex ester of L-Proline type DCK or according to any other method, usually used to carry out the reaction of a combination of amino acids.

Certain compounds of General formula (Ia), for which the R5 predstavljale as the original product using the appropriate 1-alkyl -1, -dihydro - a -(oxo or thioxo)pyridylcarbonyl acid.

Compounds of General formula (I) can be also obtained in the reaction of acylhomoserine General formula (IIb) with a dipeptide of the General formula (III) or its hydrochloride (methods B):

< / BR>
where X represents a halogen, for example chlorine, and R, R1, R2, R3 and n have the same meaning as before. When R1 has a value other than OH, the reaction is preferably carried out in the presence of organic bases such as triethylamine, whereas in the case where R1 represents OH, use inorganic bases, for example hydrates, oxides or carbonates of alkali metals or mixtures thereof.

In the case when the source of the dipeptide of the General formula (III) is a free carboxyl group and used an organic base (methods B-a), the reaction is usually carried out in a suitable nonpolar solvent, for example chloroform, methylene chloride or dioxane. When specified dipeptide contains a free carboxyl group (methods B-b ), it is preferable to use a two-phase system. The specified two-phase system usually contains an aqueous solution of an inorganic base and a solution of Azilal

When the starting compound of General formula (IIa) or (IIb) have two vicinal groups COOH or derivatives of these groups, i.e., when R represents a COOH or COOR4, COOH or COX occupy position 3, the methods A and B can lead to the formation of cyclic imides, if reaction conditions (temperature, time of reaction) are selected in such a way that they favor the intramolecular cyclization reaction products. Preferred are mild reaction conditions, especially low temperatures, because they minimize the appearance of by-products of the cyclization.

As in the case of method A, when method B is used for the dipeptide of the General formula (III), where R2 represents an amino-lower alkyl group, specified additional amino group must be previously protected by an easily removable group.

Method B (a or b) can also be used to obtain compounds of General formula (Ia) wherein R5 is a lower alkyl. In this case, the dipeptide of the General formula (III) enter into reaction with the corresponding 1-alkyl-1, -dihydro - a -(oxo - or thioxo) pyridinecarboxamide.

Specific representatives of the subgroup N-( a-mercaptopyridine is in which A is a sulfur and R5 represents hydrogen), can also be obtained in high yields by the method C, which involves the heating of compounds of General formula (I) in which R represents halogen, in the presence of sodium thiosulfate in a suitable aqueous-alcoholic medium, for example in mixtures of water and 1,2-propylene glycol.

In the case of any of the above methods A-C, when the new compounds according to the present invention receives as monoamino (prolinnova residue) or diesters (alpha-substituted on the pyridine ring carboxylic ester group), i.e. compounds of General formula (I) in which either only one COR1, or COR1, and R is an ester group of carboxylic acid, and these compounds can be converted into the corresponding free mono - or dicarboxylic acid by hydrolysis, for example, a hydrate of oxide of an alkali metal in a polar environment. Typical hydrolytic conditions include the use of potassium hydroxide, dissolved either in itself lower aliphatic alcohol (for example, containing from 1 to 3 carbon atoms), or in mixtures thereof with water.

The preferred alcohol is ethanol.

Pyridineboronic acid of General formula HN in accordance with well-known synthetic methods. The acid halides of General formula (IIb) can be easily obtained according to standard methods from the corresponding acids of General formula (IIa).

Source dipeptides of General formula (III) are also industrial products or may be synthesized according to the methods used in the chemistry of peptides.

Specific variants according to the present invention are illustrated below by examples, not limiting the invention.

The NMR spectra of H-1 and C-13 were recorded, respectively, at 199,975 and 50,289 MHz, respectively, on the spectrophotometer Varian XP-200". The values of chemical shifts are given in units of "b" on tetramethylsilane, used as internal standard. Analysis by thin-layer chromatography was performed using plates with silica gel Merck 60 F-254 with pre-coated, stain were recorded using UV-radiation. In thin-layer chromatography (TLC) used the following solvents: a:ethyl acetate, B: a mixture of ethyl acetate-acetone 3:1;With: acetone, D: absolute ethanol, E: acetic acid in ethanol, 3% F: acetic acid in ethanol, 5% of a particular type of solvent given in the parentheses in ka is inania 1).

A solution of the hydrochloride of the ethyl ester of L-alanyl-L-Proline (6 g, 0,024 mol) and triethylamine (7.4 ml) in anhydrous methylene chloride (120 ml) is cooled in a bath with ice. To the solution is added dropwise with stirring a solution of 6-chloro-2-pyridinecarboxamide (5,1 g 0,029 mol) in 30 ml of anhydrous methylene chloride. After the introduction of the components of the solution stirred for 3 h at room temperature and diluted with the reaction mixture of 400 ml of methylene chloride. The resulting solution was washed with three portions of 200 ml of 10% aqueous sodium bicarbonate solution and twice with portions of 200 ml of water. The organic layer is dried over anhydrous magnesium sulfate and the solvent is distilled off under reduced pressure, getting listed in title product as a colourless oil (yield 99%).

Range of NMR H1 (CDCl3): 1,24 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether) 1,47 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2of Proline), 3,70 (multiplet, 2H, N-CH2of Proline), is 4.15 (Quartet, J 7.2 Hz, 2H3CH2from ethyl ether) 4,50 (multiplet), 1H, CH of Proline), 4,90 (multiplet, 1H, CH of alanine, turns into a Quartet with J 6.8 Hz after shaking with deuterated water), 7,40 (double doublet, aromatic), of 8.50 (broad doublet, 1H, NH, disappears after shaking with deuterated water).

An NMR spectrum of C-13 (CDCl3): 13,9 (CH3ethyl ether) 17,7 (CH3alanine, 1 24,8 (N-CH2-CH2Proline), 28,8 (N-CH2-CH2-CH2Proline), to 46.7 (CH of alanine and N-CH2Proline), of 58.9 (CH Proline), 61,1 (CH ethyl ester), 120,9 (aromatic C-3), 127,2 (aromatic C-5), 139,9 (aromatic C-4), 150,2 (aromatic), 150,4(aroma) 162,5 (CO), 171,0 (CO), 172,1 (CO).

Thin layer chromatography (B): Rf 0,46.

Similarly synthesize the following compounds: ethyl ester of N-[(2-chloro-3-pyridinyl)carbonyl]-L-alanine-L-Proline (compound 2).

Range of NMR H1 (CDCl3): 1,28(triplet, J 7.2 Hz, 3H, CH3from ethyl ether), and 1.54 (doublet J 6.8 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2of Proline), to 3.73 (multiplet 2H, N-CH2of Proline), 4,19 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,54 (multiplet, 1H, CH of Proline), 4,96 (multiplet, 1H, CH of alanine), 7,34 (double doublet, J1 and 7.7 Hz, J2 and 4.8 Hz, 1H, aromatic C-5), 7,55 ( broad doublet, 1H, NH), 8,05 (double doublet, J1 and 7.7 Hz, J2 2 Hz, 1H, aromatic C-4), of 8.47 (double doublet, J1 4,8 Hz, J2 2 Hz, 1H, aromatic C-6).

Thin layer chromatography (B): Rf of 0.37.

B>): 1,27 (triplet J 7.2 Hz, 3H, CH3from ethyl ether) 1,50 (doublet, J 6.9 Hz, 3H, CH3from alanine), 2,10 (multiplet,4H, N-CH2-CH2-CH2of Proline), of 3.77 (multiplet with 2H, N-CH2of Proline), 4,22 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether), 4,57 (multiplet, 1H, CH of Proline), 4,91 (multiplet, 1H, CH of alanine), 7,51 (double doublet, J1 5,1 Hz, J2 1 Hz, 1H, aromatic C-5), 7,66 (doublet, J 1 Hz, 1H, aromatic C-3), 8,09 (doublet, J 1 Hz, 1H, NH), 8,43 (doublet, J 5.1 Hz, 1H, aromatic C-6).

Thin layer chromatography (B): Rf of 0.47.

Ethyl ester of N-[(6-chloro-3-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 4).

Range of NMR H1 (CDCl3): 1,27 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether) to 1.48 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,15 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,70 (multiplet, 2H, N-CH2of Proline), 4,17 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,54 (multiplet, 1H, CH of Proline), 4,94 (multiplet, 1H, CH of alanine), 7,32 (doublet, J and 8.4 Hz, 1H, aromatic C-5), 8,03 (double doublet J1 and 8.4 Hz, J2 and 2.4 Hz, 1H, aromatic C-4), 8,78 (doublet, J 2.4 Hz, 1H, aromatic C-2).

Thin layer chromatography (B): Rf 0,46.

Example 2. Synthesis of ethyl ester of N-[(1,2-dihydro-2-thioxo-4-pyridinyl)-carbonyl]-L-alanyl-L-padeniye 3) (4.5 g, of 0.013 mole) in 45 ml of a mixture of 10:1 1,2-propylene glycol: water added 15.2 g of sodium thiosulfate solution and the resulting mixture is heated at boiling under reflux for 15 hours the reaction mixture is diluted with 100 ml of water and extracted with 4 portions of 100 ml of methylene chloride. The organic layer is washed three times with 100 ml of water, dried over anhydrous magnesium sulfate and the solvent is distilled off under reduced pressure. The resulting residue is purified by crystallization from a mixture of acetone:petroleum ether, getting listed in title product in the form of hard yellow (yield 85%).

Range of NMR H1 (CDCl3): 1,27 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether) 1,68 (doublet, J 7.0 Hz, 3H, CH3from alanine), 2,10 (multiplet,4H, N-CH2-CH2-CH2of Proline in), 3.75 (multiplet, 2H, N-CH2of Proline), 4,28 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,55 (multiplet, 1H, CH of Proline), and 4.75 (multiplet, 1H, CH of alanine, turns into a Quartet with J 6.8 Hz, after shaking with deuterated water), 6,78 (double doublet, J1 and 6.6 Hz, J2 and 1.6 Hz, 1H, aromatic C-5), 7,31 (doublet, J 6.6 Hz, 1H, aromatic C-6), 7,88 (doublet, J 166 Hz, 1H, aromatic C-3), to 8.70 (broad doublet, 1H, amide NH, disappears after shaking with deuterated water is UB>2Proline), or 28.7 (N-CH2-CH2-CH2Proline) and 46.8 (N-CH Proline), 48,1 (CH alanine), a 59.2 (CH Proline), 61,2 (CH ethyl ester), 111,8 (aromatic C-5), 130,9 (aromatic C-3), 137,1 (aromatic C-6), 139,7 (aromatic C-4), 165,0 (CO), 171,7 (CO), 172,9 (CO), 178,7 (CS).

Thin layer chromatography (Sec): Rf0,50.

Similarly synthesize the following compounds.

Ethyl ester of N-[(1,2-dihydro-2-thioxo-3-pyridinyl)-carbonyl] -L-alanyl-L-Proline (compound 6).

Range of NMR H1 (CDCl3): 1,21 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether), and 1.54 (doublet, J 6.9 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline in), 3.75 (multiplet 2H, N-CH2of Proline), 4,12 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,55 (multiplet, 1H, CH of Proline), 4,85 (multiplet, 1H, CH of alanine), 6,72 (double doublet, J1 6,1 Hz, J2 and 7.6 Hz, 1H, aromatic C-5), 7,65 (double doublet J1 6,1 Hz, J2 18.7 Hz, 1H, aromatic C-4), 8,46 (double doublet J1 and 7.6 Hz, J2 and 1.7 Hz, 1H, aromatic C-6), 11,150 (broad doublet, 1H, NH).

Thin layer chromatography (Sec): Rf of 0.47.

Ethyl ester of N-[(1,2-dihydro-6-thioxo-3-pyridinyl)-carbonyl] -L-alanyl-L-Proline (compound 7).

Range of NMR H1 (CDCl3): 1,17(triplet, J 7.2 Hz, 3H, CH3from Adilova,72 (multiplet, 2H, N-CH2of Proline), 4,13 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,50 (multiplet, 1H, CH of Proline), 4,72 (multiplet, 1H, CH of alanine), 7,19 (doublet, J= 9.0 Hz, 1H, aromatic C-5), 7,56 (double doublet, J1 9,0 Hz, J2 1 Hz, 1H, aromatic C-4), 7,83 (doublet, J 2 Hz, 1H, aromatic C-2), of 8.27 (broad doublet, 1H, NH).

Thin layer chromatography (B): Rf of 0.55.

Example 3. Synthesis of ethyl ester of N-[(2-etoxycarbonyl-2-pyridinyl)-carbonyl]-L-alanyl-L-Proline (compound 8).

To the solution or 4.31 g(0,022 mol) 6-etoxycarbonyl-2-pyridineboronic acid in 100 ml of anhydrous pyridine are added during the mixing of 5.5 g (0,022 mol) of the hydrochloride of the ethyl ester of L-alanyl-L-Proline and 4.6 g of N,N'-dicyclohexylcarbodiimide. The solution is stirred for 20 min at room temperature, the volume balance dicyclohexylamine filtered and washed with acetone. The solvent from the filtrate and the wash fraction is distilled off under reduced pressure and the crude product purified by chromatographytandem on a column of silica gel using a mixture of chloroform: acetone (10:1) as eluent. Selected solid product is crystallized from a mixture of acetone-isopropyl ether-petroleum ether, receiving 4,27 g specified in the C, 3H, CH3of the aliphatic ester), 1,46 ( doublet, J 7.2 Hz, 3H, CH3from aromatic ether), 1.55V (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,15 (multiplet,4H, N-CH2-CH2-CH2of Proline in), 3.75 (multiplet, 2H, N-CH2of Proline), 4,21 (Quartet, J 7.2 Hz, 2H, CH2of the aliphatic ether complex), 4,48 (Quartet, J 7.2 Hz, 2H, CH2of complex aromatic ether), 4,53 (multiplet, 1H, CH of Proline), 5,00 (multiplet, 1H, CH of alanine, turns into a Quartet with J 6.8 Hz after shaking with deuterated water), 7,99 ( double doublet, J1 J2 7,6 Hz, 1H, aromatic C-4), by 8.22 (double doublet, J1 7 Hz, J2 1 Hz, 1H, aromatic), 8,33 (double doublet, J1= 7,6 Hz, J2 1 Hz, 1H, aromatic), to 8.70 (broad doublet, 1H, amide NH, disappears after shaking with deuterated water).

An NMR spectrum of C-13(CDCl3): 14,1 (CH3ethyl ether) 14,2 (CH ethyl ether) 17,8 (CH3alanine), 24,9 (N-CH2-CH2from Proline) 29,0 (N-CH2-CH2-CH2Proline), 46,9 (CH of alanine and N-CH2Proline), 59,3 (CH Proline), 61,3 (CH ethyl ether) of 62.4 (CH2ethyl ether), output reached 125.5(aromatic, uniforms, 127.6 (aromatic), 138, 7mm (aromatic C-4), 147,6 (aromatic), to 150.1 (aromatic), 163,5 (CO), 115,0 (CO), OF 171.2 (CO), 172,4 (CO)

Thin layer chromatography (B): Rf 0,63.

Range of NMR H1 (CDCl3): 1,26 (triplet, J 7.2 Hz, 3H, CH3of aliphatic ethyl ester) and 1.51 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,80 (multiplet, 2H, N-CH2of Proline), a 4.03 (singlet, 3H, CH3aromatic methyl ether complex), 4,217 (Quartet, J 7.2 Hz, 2H, CH of ethyl ether) 4,55 (multiplet, 1H, CH of Proline), 4,99 (multiplet, 1H, CH of alanine), 8,12 (doublet, J and 8.4 Hz, 1H, aromatic C-3), compared to 8.26 (double doublet, J1 and 8.4 Hz, J2 2 Hz, 1H, aromatic C-4), 8,43 (doublet, J 7.4 Hz, 1H, NH), 9,12 (doublet, J 2.0 Hz, 1H, aromatic C-6).

Thin layer chromatography (B): Rf 0,36.

Ethyl ester of N-[(2-methoxycarbonyl-3-pyridinyl)-carbonyl]-L-alanyl-L-Proline (compound 10).

Range of NMR H1 (CDCl3): 1.23 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether) 1,50 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,10 (multiplet,4H, N-CH2-CH2-CH2of Proline), 3,70 (multiplet, 2H, N-CH2of Proline), 3,94 (singlet, 3H, CH2methyl ether complex), is 4.15 (Quartet, J 7.2 Hz, 2H, CH of ethyl ester), 4,50 (multiplet, 1H, CH of Proline), 4,80 (multiplet, 1H, CH of alanine), 7,0 (broad doublet, 1H, NH), 7,45 (double doublet, J1 and 7.8 Hz, J2 of 4.7 Hz, 1H, aromatic C-5), 7,84 (double doublet, J1 and 7.8 Hz, J2 and 1.7 Hz, 1H, ar is ptx2">

Thin layer chromatography (B): Rf 0,27.

Ethyl ester of N-[(5-bromo-1,2-dihydro-2-oxo-3-pyridinyl)-carbonyl]-L-alanyl - L-Proline (compound 11).

Range of NMR H1 (CDCl3): 1,28 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether) 1,53 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline in), 3.75 (multiplet, 2H, N-CH2of Proline), 4,20 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,60 (multiplet, 1H, CH of Proline), 4,88(multiplet, 1H, CH of alanine), 7,71 (doublet, J 2.8 Hz, 1H, aromatic C-4), 8,48 (doublet, J 2.8 Hz, 1H, aromatic C-6), 10,10 (broad doublet, 1H, amide NH).

Thin layer chromatography (B): Rf 0,76.

Ethyl ester of N-[(1,2-dihydro-2-oxo-4-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 12).

Range of NMR H1 (CDCl3): 1,21 (triplet, J= 7.2 Hz, 3H, CH3from ethyl ether) 1,47 (doublet, J 7.0 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline in), 3.75 (multiplet, 2H, N-CH2of Proline), 4,12 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,50 (multiplet, 1H, CH of Proline), 4,80 (multiplet, 1H, CH of alanine), 6,51 (double doublet, J1= 6,7 Hz, J2 1.2 Hz, 1H, aromatic C-5), 6,97 (doublet, J 1.2 Hz, 1H, aromatic C-3), 7,25 (doublet, J 6,7 Hz, 1H, aromatics the Il-N-[(l,2-dihydro-6-oxo-2-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 13).

Range of NMR H1 (CDCl3): 1,22 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether) to 1.48 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,10 (multiplet,4H, N-CH2-CH2-CH2of Proline), 3,57 (multiplet, 2H, N-CH2of Proline), 4,17 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether), 4,48 (multiplet, 1H, CH of Proline) 4,90 (multiplet, 1H, CH of alanine), 6.73 x (double doublet, j-1 and 9.2 Hz, J2 0.8 Hz, 1H, aromatic C-5), 6,91 (double doublet, J1 7,0 Hz, J2 1.0 Hz, 1H, aromatic C-3), 7,44 (double doublet, J1 9,0 Hz, J2 7,0 Hz, 1H, aromatic C-4), 8,46 (broad doublet, 1H, amide NH).

Thin layer chromatography (Sec): Rf of 0.43.

Ethyl ester of N-[(1,2-dihydro-6-oxo-3-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 14).

Range of NMR H1 (CDCl3): 1,22 (triplet, J 7.0 Hz, 3H, CH3from ethyl ether), 1,40 (doublet, J 7.0 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline in), 3.75 (multiplet, 2H, N-CH2of Proline), 4,10 (Quartet, J 7.0 Hz, 2H, CH2from ethyl ether) 4,50 (multiplet, 1H, CH of Proline), 4,85 (multiplet, 1H, CH of alanine), 6,34 (doublet, J 9.0 Hz, 1H, aromatic C-5), 7,80 (doublet, J 8,8 Hz, 1H, aromatic C-4), 7,97 (singlet, 1H, aromatic C-2), 8,65 (doublet, J 6.8 Hz, amide NH).

Thin layer chromatography (D): Rf 0.50 in.

3): to 1.15 (triplet, J and 7.1 Hz, 3H, CH3from ethyl ether) of 1.30 (doublet, J 7.0 Hz, 3H, CH3from alanine), 2,10 (multiplet,4H, N-CH2-CH2-CH2of Proline), 4,30 (multiplet, 2H, N-CH2of Proline), 4,05 (Quartet, J 7,1 Hz, 2H, CH2from ethyl ether) 4,33 (multiplet, 1H, CH of Proline), 4,80 (multiplet, 1H, CH of alanine), 7,21 (multiplet, 5H, O-phenyl), 7,42 (multiplet, 2H, aromatic), 8,23 (multiplet, 2H, aromatic), 8,78 (doublet, J 8.0 Hz, 1H, amide NH).

Thin layer chromatography: Rf 0,39.

Ethyl ester of N-[(2-phenylthio-3-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 16).

Range of NMR H1 (CDCl3): 1,16 (triplet J 7.2 Hz, 3H, CH3from ethyl ether), 1,32 (doublet, J 7.0 Hz, 3H, CH3from alanine), 1,96 ( multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,69 (multiplet, 2H, N-CH2of Proline), 4,06 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether), 4,35 (multiplet, 1H, CH of Proline), 4,72 (multiplet, 1H, CH of alanine), 7,21 (double doublet, J1 8.0 Hz, J2 and 4.8 Hz, 1H, aromatic C-5), 7,88 (double doublet, j-1 and 7.6 Hz, J2 1.8 Hz, 1H, aromatic C-4), 8,33 (double doublet, J1 4,8 Hz, J2 1.8 Hz, 1H, aromatic C-6), 8,87 (doublet, J1 8.0 Hz, 1H, amide NH).

Thin layer chromatography (A) Rf of 0.33.

Ethyl ester of N-[(2-phenylamino-3-pyridine is of the ethyl ester), 1,19 (doublet, J 6.6 Hz,3H, CH3from alanine), 1,72 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,70 (multiplet, 2H, N-CH2of Proline), a 4.03 (Quartet, J 7.2 Hz, 2H, CH2from ethyl ether) 4,20 (multiplet, 1H, CH of Proline), 4,56 (multiplet, 1H, CH of alanine), 7,22 (multiplet, 5H, aromatic), 7,45 (double doublet J1 6,0 Hz, J2 and 4.6 Hz, 1H, aromatic C-5), 7,60 (doublet, J 8.0 Hz, 1H, 11H), 8,08 (multiplet, 1H, aromatic), of 8.47 (double doublet, J1 4,8 Hz, J2 and 1.4 Hz, 1H, aromatic C-6), 8,91 (singlet, 1H, amide NH).

Thin layer chromatography (A) Rf 0,22.

Ethyl ester of N-[(1,2-dihydro-5-nitro-2-oxo-3-pyridinyl)carbonyl]-L-alanyl - L-Proline (compound 18).

Range of NMR H1 (CDCl3): 1,17 (triplet, J and 7.1 Hz, 3H, CH3from ethyl ether) 1,29 (doublet, J 7.0 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,60 (multiplet, 2H, N-CH2of Proline), 4,12 (Quartet, J 7,1 Hz, 2H, CH2from ethyl ether), 4,32 (multiplet, 1H, CH of Proline), 4,78 (multiplet, 1H, CH of alanine), 8,77 (doublet, J 3.3 Hz, 1H, aromatic C-4), 8,89 (doublet, J3,3 Hz, 1H, aromatic C-6), 10,01 (doublet, J 6.9 Hz, 1H, amide NH).

Thin layer chromatography (D): Rf 0,74.

Ethyl ester of N-[(l,2-dihydro-1-methyl-2-oxo-3-pyridinyl)-carbonyl]-L-alanyl - is), 1,28 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,10 ( multiplet,4H, N-CH2-CH2-CH2of Proline), 3,45 (singlet, 3H, N-CH), 3,60 (multiplet, 2H, N-CH2of Proline), 4,06 (Quartet, J 7,2 H Hz, 2H, CH2from ethyl ether), or 4.31 (multiplet, 1H, CH of Proline), 4,76 (multiplet, 1H, CH of alanine), 6,48 (double doublet J1 J2 7,0 Hz, 1H, aromatic C-5), of 8.06 (double doublet, J1 7,0 Hz, J2 2,2 Hz, 1H, aromatic C-4), 8,29 (double doublet, J1= 7.0 Hz, J2 2,2 Hz, 1H, aromatic C-6), 10,23 (broad doublet, 1H, NH).

Thin layer chromatography (D): Rf 0,51.

Ethyl ester of N-[(2-acetylamino-4-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 20).

Range of NMR H1 (DMSO-d6): 1,17 (triplet, J 7.5 Hz, 3H, CH3from ethyl ether), 1,33 (doublet, J 7.0 Hz, 3H, CH3from alanine), 1,96 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 2,11 (singlet, 3H, CH3from acetylamino), 3,67 (multiplet, 2H, N-CH2of Proline), 4,07 (Quartet, J 7.5 Hz, 2H, CH2from ethyl ether) 4,33 (multiplet, 1H, CH of Proline), 4,72 (multiplet, 1H, CH of alanine), 7,49 (doublet, J 5 Hz, 1H, aromatic C-5), to 8.41 (singlet, 1H, aromatic C-3), 8,43 (doublet, J 5 Hz, 1H, aromatic C-6), 8,91 (broad doublet, 1H, NH, disappears after shaking with deuterated water), 10,66 (broad doublet, 1H, NH from acetylamino the ASS="ptx2">

Example 4. Synthesis of ethyl ester of N-[(l,2-dihydro-2-oxo-3-pyridinyl)-carbonyl]-L-alanyl-L-Proline (compound 21).

To a solution of the hydrochloride of the ethyl ester of L-alanyl-L-Proline (4.0 g, to 0.016 mol) and triethylamine (8 ml) in 200 ml of dioxane is added dropwise over 1 h with stirring, maintaining a room temperature, a suspension of 3.0 g (0.019 mol) 2-oxycodoneshare in 200 ml of anhydrous dioxane. After stirring the mixture for an additional 3 h, the solvent is distilled off under reduced pressure. Thus obtained crude product is dissolved in 500 ml of chloroform and the solution washed twice with 100 ml 5% aqueous sodium carbonate solution. The organic layer is dried over anhydrous magnesium sulfate and remove the solvent under reduced pressure, obtaining specified in the header on a rotary evaporator. The solid residue is crystallized from a mixture of methylene chloride and isopropyl ether, obtaining 3.4 g specified in the connection header in the form of a microcrystalline solid product (yield 64%).

Range of NMR H1 (CDCl3): 1,23 (triplet, J 7.2 Hz, 3H, CH3from ethyl ether) 1,47 (doublet, J 6.8 Hz, 3H, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,70 (mult is a), 4,92 (multiplet, 1H, CH of alanine,turns into a Quartet c J 6.8 Hz, after shaking with deuterated water), 6,44 (double doublet, J1 7,4 Hz, J2 6.2 Hz, 1H, aromatic C-5), 7,65 (double doublet, J1 6,2 Hz, J2 2,2 Hz, 1H, aromatic C-4), 8,48 (double doublet, J1 7,4 Hz, J2 2,2 Hz, 1H, aromatic C-6), and 10.20 (broad doublet, 1H, amide NH, disappears after shaking with deuterated water).

An NMR spectrum of C-13 (CDCl3): 13,9 (CH3ethyl ether) to 17.6 (CH3alanine), 24,7 (N-CH2-CH2Proline), 28,8 (N-CH2-CH2-CH2Proline), 46,9 (N-CH2of Proline), 47,1 (CH alanine), of 58.9 (CH Proline), 61,0 (CH2ethyl ether), 107,4 (aromatic C-5), 120,9 (aromatic C-3), 138,9 (aromatic C-6), RUB 145.1 (aromatic C-4), 163,4 (CO), 163,9 (CO), 171,7 (CO), 172,2 (CO).

Thin layer chromatography (Sec): Rf 0,36.

Example 5. Synthesis of N-[(6-chloro-3-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 22).

1.0 g (0.54 mmol) of L-alanyl-L-Proline dissolved in a mixture with 10.8 ml of 0.5 H a solution of potassium hydroxide and 750 ml (5.4 mmol) of anhydrous potassium carbonate and added to a solution of 10 ml of acetonitrile. After cooling in a bath with ice to the solution is added dropwise with stirring a solution of 1.2 g of 6-chloro-nicotinanilide, maintaining the pH of the mixture in the range from 12 to 13 is now the solution is stirred for 2 h at room temperature, then neutralized to pH 6 by means of an aqueous solution of hydrochloric acid and the solvent is distilled off under reduced pressure. Thus obtained residue was transferred to a suspension in 50 ml of absolute ethanol, the residue of potassium chloride is separated by centrifugation and evaporated from a transparent solution of the solvent under reduced pressure. The crude product is purified by chromatographytandem on a column of silica gel, using as eluent ethanol, receiving specified in the header of the product in the amount of 700 mg (yield 40%).

Range of NMR H1 (DMSO + D2O): 1,26, 1,32 (two doublet, J 6.8 Hz, CH3alanine), 1,90 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,50 (multiplet, 2H, N-CH2of Proline), 4,15 (multiplet, 1H, CH of Proline), 4,70 (multiplet, 1H, CH of alanine), 7.62mm (double doublet, J1 8,2 Hz, J2 and 3.0 Hz, 1H, aromatic C-5), 8,32 (double doublet, J1 8,2 Hz, J2 1 Hz, 1H, aromatic C-4), 8,87 (doublet, J 3.0 Hz, 1H, aromatic C-2).

An NMR spectrum of s-13 (DMSO + D2O): 16,6, 18,1 (CH3alanine), 22,3, 24,78 (N-CH2-CH2of Proline), 29,4, 31,7 (N-CH2-CH2-CH2Proline), 46,847,6, of 48.7 (CH of alanine and N-CH2Proline), 61,161,9 (CH Proline), of 124.8 (aromatic C-5), of 129.6 (aromatic C-3), 139,6 (aromatic C-4), 149,9 (aroma is), covers 175.6 (COOH Proline).

Thin layer chromatography (E.): Rf 0,54.

Similarly synthesize the following compounds.N-[(2-methoxycarbonyl-5-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 23).

Range of NMR H1 (DMSO + D2O): 1,26, 132,32 (two doublet, J 6.8 Hz, CH of alanine), 1,90 (multiplet, 4H, N-CH-CH-CH of Proline), 3,50 (multiplet, 2H, N-CH of Proline), 4,15 (multiplet, 1H, CH of Proline), 4,70 (multiplet, 1H, CH of alanine), 7.62mm (double doublet, J1 8,2 Hz, J2 and 3.0 Hz, 1H, aromatic C-5), 8,32 (double doublet, J1 8,2 Hz, J2 1.0 Hz, 1H, aromatic C-4), 8,87 (doublet, J 3.0 Hz, 1H, aromatic C-2).

Thin layer chromatography (D): Rf 0.35 in.

Example 6. Synthesis of N-[(6-chloro-2-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 24).

A solution of the hydrochloride of the ethyl ester of L-alanyl-L-Proline (4.5 g, of 0.013 mole) ( example 1) in 42 ml of an ethanol solution of potassium hydroxide is stirred at room temperature for 3 hours the reaction mixture is diluted with 250 ml absolute ethanol and neutralized 6N ethanolic hydrochloric acid. The resulting precipitate of potassium chloride is filtered off, washed several times with absolute ethanol and the solvent is distilled off under reduced pressure. The residue is subjected to by the title product as a colorless solid (yield 73%).

Range of NMR H1 (DMSO): 1,36, 1,32 (two doublet, J= 6,8 Hz, CH3from alanine), 2,10 (multiplet, 4H, N-CH2-CH2-CH2of Proline), 3,65 (multiplet, 2H, N-CH2of Proline), 4.30 multiplet, 1H, CH of Proline), 4,51, 4,75 (two multiplet, 1H, CH of alanine), 7,78 (double doublet, J1 7,4 Hz, J2 and 1.6 Hz, 1H, aromatic), 8,03 (double doublet, J1 7,4 Hz, J2 and 1.6 Hz, 1H, aromatic), 8,10 (double doublet, J1 J2 7,4 Hz, 1H, aromatic C-4), to 8.70 (broad doublet, 1H, amide NH, disappears after shaking with deuterated water).

An NMR spectrum of C-13 (DMSO): 17,3, 18,6 (CH3alanine), 21,8, 24,6 (N-CH2-CH2Proline), 28,6, 30,8 (N-CH2-CH2-CH2Proline), 46,3, 46,546,7 and 46.8 (CH of alanine and N-CH2Proline), 58,7, 59,3 (CH Proline), 121,5 (aromatic C-3), 127,8 (aromatic C-5), 141,9 (aromatic C-4), 149,6 (aromatic), 150,4 (aromatic), 161,8 (CO amide bond pyridine ring-dipeptide), 170,3, to 170.9 (CO-peptide bond), 173,8 (COOH Proline).

Thin layer chromatography (E.): Rf of 0.55.

This splitting of the signals due to the presence of z-Cys and z-TRANS peptide bond L-alanine-L-Proline.

Similarly receive the following free acids from the corresponding esters:

N-[(6-chloro-3-pyridinyl)carbonyl]-L-alanyl-L-p is Olin (compound 26), thin layer chromatography (E.): Rf of 0.55,

N-[(1,2-dihydro-2-thioxo-4-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 27), thin layer chromatography (E.): Rf 0,46.

N-[(1,2-dihydro-2-thioxo-3-pyridinyl)carbonyl] -L - alanyl-L-Proline (compound 28), thin layer chromatography (E.): Rf 0.45 in.

N-[(1,2-dihydro-6-thioxo-3-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 29), thin layer chromatography (E.): Rf 0,51.

N-[(2-carboxy-6-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 30), thin layer chromatography (D): Rf of 0.29.

N-[(2-carboxy-5-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 31), thin layer chromatography (D): Rf 0.25 in.

N-[(2-carboxy-3-pyridinyl)carbonyl] -L-alanyl-L-Proline (compound 32), thin layer chromatography (D): Rf 0,18.

N-[(1,2-dihydro-2-oxo-4-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 33), thin layer chromatography (f): Rf 0.34 in.

N-[(1,6-dihydro-6-oxo-2-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 34), thin layer chromatography (f): Rf 0.40 in.

N-[(1,6 - dihydro-6-oxo-3-pyridinyl)carbonyl] -L-Ala - Nile-L-Proline (compound 35), thin layer chromatography (f): Rf of 0.37.

N-[(l,2-dihydro-2-oxo-3-pyridinyl)carbonyl]-L-alanyl-L-Proline (compound 36), thin-layer address chronic spectroscopic data.

Example 7. The results of pharmacological tests.

A. Inhibition of contractions of the ileum from Guinea pigs induced by angiotensin I (see tab. 2).

Segments to 1.5 cm svezhenanesennoj and washed-terminal part of the ileum was transferred to a suspension of 25 mm vessels to tissues containing solution on Tirade at the 31oC, and passed through a suspension of a mixture of 95% O2and 5% CO2. The residual tension of the fabric after equilibration and feedback on angiotensin I or angiotensin II (both at a concentration of 100 ng/ml) during the initial load of 1.0 g register by means of the receiver-Converter "Ealing", and multichannel recorder "Electromed". Part of the compounds according to the present invention is tested in accordance with the described method, adding them to the vessel 2 min prior to entering the appropriate tools-agonist.

The inhibitory effect of the tested compounds in relation to induced angiotensin I reductions determined at final concentration in the vessel 10 ng/ml

The results are shown in table. 1.

All are values of percentage inhibition represent average values obtained on Segni one of the tested compounds does not give any significant changes reductions caused by angiotensin II.

B. Protivogipertonicheskoe activity when tested on rats with spontaneous pressure increase (table. 3 and 4).

In this well-known model of hypertension using rats (weighing from 315 to 376 g) of the spontaneous increase in pressure (systolic blood pressure (ACS), more than 160 mm RT.cent.). Systolic blood pressure recorded in the consciousness of animals by the method using the tail cuff and device for measuring pressure of 40 FS. Before measurements, the rats are placed in an insulated cylinder, register initial values of pressure and injected them with compound (including used as a comparative sample captopril) in oral doses of 30 mg/kg (using 5 animals per group). The pressure is measured after 1,2,3,4 and 24 h after injection of the tested compounds. The control group (18 animals) include in the experiment for comparison (table. 2).

Under the above conditions the connection 8, 27, and 21 demonstrate the significant protivogipertonicheskoe effect. After 1 h after injection, the average decrease in systolic blood pressure is significantly different from the control group for groups, about Eisenia pressure is significantly different from the control group after 2, 34 and 24 h in the case of compounds 8 and 27, whereas captopril becomes after 24 h inactive (according to test Duncan-Kramer p<0,05). The average initial values of systolic blood pressure (of 167.2 to 169,0 mm RT.CT.) a little different from the corresponding values in the control group (167,8 0,61 mm RT.cent.).

Connection 21 at a concentration of 30 mg/kg shows significant decrease pressure only after 2 hours, at a dose of 45 mg/kg lowered blood pressure significantly also after 2, 3, 4, and 24 h Protivogipertonicheskoe effect in accordance with the above described model is also observed by oral administration of compounds 8 and 27 in doses of 7.5 and 15 kg/kg.

Supplement V.

Synthesis of sodium salt of N-[(2-chloro-4-pyridyl)carbonyl]-L-alanyl-L-Proline.

A mixture of 29.4 g (0.09 mol) of N-[(2-chloro-4-pyridyl)carbonyl]-L-alanyl-L-Proline and 3.85 g (0.09 mol) of sodium hydroxide (99%) in 300 ml of absolute ethanol is stirred at room temperature until complete dissolution (1 h).

The solvent is evaporated in vacuum. The residue is dissolved in 200 ml of acetone, are planted diethyl ether to obtain the titled compound as a colorless solid precipitate.

1H-NMR (DMSO): 1,26, 1,31 (two d, J 7 Hz, CH3Alan is on), the 7.85 (m, 1H, aromatic), to 7.95 (m, 1H, aromatic), 8,53 (m, 1H, aromatic), 8,98, which 9.22 (two Shire. D. 1H, amide NH).

13C-NMR (DMSO)": 13,3, 17,3 (CH3alanine), 22,0, 24,2 (N-CH2-CH2Proline), 29,1, 31,3 (N-CH2-CH2-CH2Proline), 48,2, 46,3, 47,3, 47,7 (N-CH2Proline and CH of alanine), 61,4, 31,7 (CH Proline), to 121.5 (C-5 aromatic), 122,5, the 122.7 (C-3 aromatic), 144,8, 144,9 (C-4 aromatic) 150,9, 151,0, 151,1, 151,2 (C-2, C-6, aromatic), 133,0, 133,2 (CO amide bond pyridine ring of the dipeptide), 139,8, 170,3 (CO peptide bond), 175,5, 176,1 (COO-Proline).

The doubleness of the signal is due to the presence of isomers z-CIS and z-TRANS by peptide bonds L-Ala-L-Pro.

Annex VI.

The study of the toxicity of selected compounds (PCT/EP 92/00400; EN Appl N 92 016 314.04)

Study of acute toxicity is held to some of the most active compounds in vivo (HP test, see Appendix II), especially for N 2, 3, 8, 15, 27, as for N 21, the most active compound in vitro (Test of Angiotensin I, see Appendix I).

Acute toxicity in mice.

Assessment of LD50on the 7th day of observation gives the following values presented in table.5.

Specific learning connection N 3.

A) Comparative observation CLM (maximum tolerated dose) was 5000 mg/kg (p. O.) and 2000 mg/kg (b).

B) Subacute toxicity in rats, when introduced through the mouth (R. O.).

The connection 3 is administered for 28 days at a dose of 0 (control), 500, 1000, 2000 and 4000 mg/kg/day.

Not see significant signs of toxicity and no deaths, but all groups had similar weight gain.

In samples of liver and kidney histological studies have not detected changes in any doses.

Supplement VII.

Farbkomposition.

Because the oral route is the preferred route of administration, and because many of these new compounds are active or very active in this introduction, pharmaceutical forms for oral administration are chosen as target in the development of herbal medicines. From the analysis of different preparative forms some specific examples are included here. They are only intended to illustrate, without limitation, in any case, the field of invention.

Preparative form in tablet form.

When used as an active ingredient the compound N 3, the chosen representative of these protivogipertonicheskoe dipeptide derivatives are given the following formulation or p is ava and techniques

Preparative form A mg/tablet

a) active ingredient 100

b) avicel PH-102 100

c) doonally calcium phosphate 74

d) Ac-Di-Sol 7.4

e) talc 15

f) magnesium stearate 3

g), Aerosil 200 0,6 300

Avicel PH-102 is a commercial microcrystalline cellulose. Ac-Di-Sol is a commercial carboxymethyl cellulose. sodium (stitched). Aerosil 100 is a commercial colloidal silicon dioxide with a specific surface area of 200 m2/,

After pre-mixing ingredients d, e, f, g, with about half of the corresponding number of b and c add the active ingredient and the remnants of b and c is stirred for 30 min, followed by pressing.

Preparative form B mg/tablet

a) active ingredient 50

b) lactose 50

c) corn starch 33

d) povidone 4

e) Ac-Di-Sol 4

f) talc 7,5

g) magnesium stearate 1.5 to 150

This preparative form getting wet granulation of the ingredients a, b and c with 10% (weight/weight) solution of povidone in water, followed by drying, mixing with other ingredients and pressing.

II. Characteristics.

And form A and form B are tested for their galenovyh drugs with the corresponding values of DK are shown in table.6.

1. Derivatives of dipeptides of General formula I

< / BR>
including tautomeric forms,

where n is 0 or 1;

R is halogen, HE, SH, SR4OR4where R4lower alkyl or phenyl, with other5where R5phenyl or acetyl, di(lower alkyl)amino, COOH or COO(lower)alkyl;

R1HE or (lower)alkoxy;

R2lower alkyl;

R3halogen, NO2, (lower)alkyl, aryl(lower)alkyl,

or their pharmaceutically acceptable salts.

2. Derivatives of dipeptides of General formula I on p. 1, where n is 0, R is chlorine, HE, SH, OS6H5SC6H5, NHC6H5, NHCOCH3, COOH or COO(lower)alkyl.

3. Derivatives of dipeptides of General formula I on p. 1, where n is 1, R is chlorine, HE, co3or N(CH3)2and R3selected from halogen, in particular chlorine, methyl or phenylmethyl.

4. Derivatives of dipeptides of General formula I on PP.1 3, where R1HE or OS2H5, R2methyl.

5. A derivative of the dipeptide under item 1, which is the ethyl ester of N-[(2-chloro-4 - pyridinyl)carbonyl]-L-alanyl-L-Proline.

6. A derivative of the dipeptide under item 1, which is the ethyl ester of N-[(2-phenoxy-3 - pyridinyl)carbonyl]-L-alanyl-L-Proline.

7. A derivative of the dipeptide under item 1, judnich of dipeptides of General formula I

< / BR>
including tautomeric forms,

where n is 0 or 1;

R is halogen, HE, SH, SR4OR4where R4lower alkali or phenyl, with other5where R5phenyl or acetyl, di(lower alkyl)amino, -COOH or-COO (lower)alkyl;

R1HE or (lower)alkoxy;

R2lower alkyl;

R3halogen, NO2, (lower)alkyl, aryl(lower)alkyl,

or their pharmaceutically acceptable salts, characterized in that conduct the interaction pyridineboronic acid of formula IIa

< / BR>
where n is 0 or 1;

R is halogen, HE, SR4OR4where R4lower alkyl or phenyl, with other5where R5phenyl or acetyl, di(lower)alkylamino, -COO(lower)alkyl;

R3halogen, NO2, (lower)alkyl, aryl(lower)alkyl,

with the dipeptide of formula III

< / BR>
where R1(lower)alkoxy;

R2lower alkyl,

in the presence of a suitable agent combinations, for example N,N-dicyclohexylcarbodiimide, followed if necessary by conversion of compounds of formula I, where R is Cl, the compound I, where R is SH, and, if necessary, by hydrolysis, by salt formation fats or diesters obtained in this manner.

9. The method of producing dipeptides of General formula I

< / BR>
including Tau the NH5where R5phenyl or acetyl, di(lower alkyl)amino, COOH or COO(lower)alkyl;

R1HE or (lower)alkoxy;

R2lower alkyl;

R3halogen, NO2, (lower)alkyl, aryl(lower)alkyl,

or their pharmaceutically acceptable salt,

characterized in that a derivative of General formula II

< / BR>
where n is 0 or 1;

X halogen;

R is halogen, HE, SR4OR4where R4lower alkyl or phenyl, with other5where R5phenyl or acetyl, di(lower alkyl)amino, COOH or COO(lower)alkyl;

R3halogen, NO2, (lower)alkyl, aryl(lower)alkyl,

enter into interaction with the dipeptide of formula III

< / BR>
where R1HE or (lower)alkoxy;

R2lower alkyl,

the process is conducted in the presence of organic bases, if R1IT, the process is conducted in the presence of inorganic bases and, if necessary, with subsequent conversion of compounds of formula I, where R is Cl, the compound of formula I, where R is SH, and, if necessary, by hydrolysis and/or salt formation of ester or free acid thus obtained.

10. Antihyperlipidemia pharmaceutical composition comprising an active ingredient and pharmaceutically acceptable nosp.1 7 in an effective amount.

11. The composition according to p. 10 for oral administration.

 

Same patents:
The invention relates to a method of crystallization by cooling methyl ester of alpha-L-aspartyl-L-phenylalanine (hereinafter AWS), which is likely to find wide application as a low-calorie zacharopoulou substance because it has the sweetness of high quality and is approximately 200 times sweeter than sugar

The invention relates to the synthesis of biologically active compounds, in particular to new derivatives of N-phenylglycinate formula

< / BR>
the way they are received and to farbkomposition based on them

The invention relates to medicine, namely to compounds having immunomodulatory properties

The invention relates to medicine, particularly cardiology, and can be used for the treatment and prevention of the consequences and complications of ischemia and reperfusion injury of the heart

The invention relates to biology, immunology, biotechnology, agriculture, farming, animal husbandry

The invention relates to pharmaceutical industry

The invention relates to the synthesis of biologically active compounds, in particular to new derivatives of N-phenylglycinate formula

< / BR>
the way they are received and to farbkomposition based on them

The invention relates to new biologically active compounds derived boron-containing peptides and pharmaceutical composition having inhibitory activity to trypsinogen serine proteases, which can find application in biology and medicine

FIELD: organic chemistry, medicine.

SUBSTANCE: invention relates to applying compounds of the formula (I) for preparing an antibacterial composition and veterinary composition eliciting with the enhanced activity.

EFFECT: valuable properties of agents.

4 cl, 3 tbl, 78 ex

FIELD: organic chemistry, biochemistry, medicine, pharmacy.

SUBSTANCE: invention relates to macrocyclic peptides of the general formula (I): wherein W means nitrogen atom (N); R21 means hydrogen atom (H), (C1-C6)-alkoxy-, hydroxy-group or N-(C1-C6-alkyl)2; R22 means hydrogen atom (H), (C1-C6)-alkyl, CF3, (C1-C6)-alkoxy-group, (C2-C7)-alkoxyalkyl, C6-aryl or Het wherein het means five- or six-membered saturated or unsaturated heterocycle comprising two heteroatoms taken among nitrogen, oxygen or sulfur atom and wherein indicated Het is substituted with radical R24 wherein R23 means hydrogen atom (H), -NH-C(O)-R26, OR26, -NHC(O)-NH-R26, -NHC(O)-OR26 wherein R26 means hydrogen atom, (C1-C6)-alkyl; R3 means hydroxy-group or group of the formula -NH-R31 wherein R31 means -C(O)-R32, -C(O)-NHR32 or -C(O)-OR32 wherein R32 means (C1-C6)-alkyl or (C3-C6)-cycloalkyl; D means a saturated or unsaturated alkylene chain comprising of 5-10 carbon atoms and comprising optionally one-three heteroatoms taken independently of one another among oxygen (O), sulfur (S) atom, or N-R41 wherein R41 means hydrogen atom (H), -C(O)-R42 wherein R42 means (C1-C6)-alkyl, C6-aryl; R4 means hydrogen atom (H) or one-three substitutes at any carbon atom in chain D wherein substitutes are taken independently of one another from group comprising (C1-C6)-alkyl, hydroxyl; A means carboxylic acid or its alkyl esters or their derivatives. Invention relates to pharmaceutical compositions containing indicated compounds and eliciting activity with respect to hepatitis C virus and these peptides inhibit activity of NS3-protease specifically but don't elicit significant inhibitory activity with respect to other serine proteases.

EFFECT: valuable biochemical and medicinal properties of peptides.

106 cl, 9 tbl, 61 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to compounds of the formula (I):

wherein r = 1, 2 or 3; s = 0; t = 0; R1 is taken among group including R11-CO and R12-SO2- wherein R11 is taken among group including (C6-C14)-aryl, (C1-C8)-alkyloxy-group wherein all given group are unsubstituted or substituted with a single or some similar or different substitutes R40; R12 means (C6-C14)-aryl wherein indicated group is unsubstituted or substituted with a single or some similar or different substituted R40; R2 means R21(R22)CH-, R23-Het-(CH2)k-, R23(R24)N-(CH2)m-D-(CH2)n- or R25(R26)N-CO-(CH2)p-D-(CH2)q- wherein D means bivalent residue -C(R31)(R32)-, bivalent (C6-C14)-arylene residue or bivalent residue obtained from aromatic group Het comprising 5 or 6 atoms in cycle among them 1 or 2 are similar or different cyclic heteroatoms taken among group including nitrogen and sulfur atoms; numbers k, m, n, p and q = 0, 1, 2; R21 and R22 that are independent of one another can be similar or different and taken among group including hydrogen atom, (C1-C12)-alkyl, (C6-C14)-aryl and so on; R23 means hydrogen atom, R27-SO2- or R28-CO-; R24, R25 and R26 mean hydrogen atom; R27 is taken among group including (C1-C8)-alkyl, (C6-C14)-aryl and so on; R28 is taken among group including R27, (C1-C8)-alkyloxy-group; R31 and R32 mean hydrogen atom; R40 is taken among group including halogen atom, hydroxy-, (C1-C8)-alkyloxy-group, (C1-C8)-alkyl, (C6-C14)-aryl and so on; R91, R92, R93 and R96 means hydrogen atom; R95 means amidino-group; R97 means R99-(C1-C8)-alkyl; R99 is taken among group including hydroxycarbonyl- and (C1-C8)-alkyloxycarbonyl-; Het means saturated, partially unsaturated or aromatic monocyclic structure comprising from 3 to 6 atoms in cycle among them 1 or 2 are similar or different heteroatoms taken among group comprising nitrogen and sulfur atoms; in all its stereoisomeric forms and also their mixtures in any ratios, and its physiologically acceptable salts. Invention proposes a method for preparing compound of the formula (I). Also, invention proposes a pharmaceutical preparation eliciting inhibitory activity with respect to factor VIIA and containing at least one compound of the formula (I) and/or its physiologically acceptable salts and pharmaceutically acceptable carrier. Invention provides preparing compounds of the formula (I) eliciting power anti-thrombosis effect and useful for treatment and prophylaxis of thrombosis-embolic diseases.

EFFECT: valuable medicinal properties of compounds and composition.

10 cl, 70 ex

FIELD: organic chemistry and drugs.

SUBSTANCE: New class of compounds of general formula 1, where R has formula 2 or 3; other residues are as described in claim of invention is disclosed. Said compounds are interleikyn-1β converting enzyme (ICE) inhibitors and have specific structural and physicochemical properties. Invention also relates to pharmaceutical composition containing said compounds. Compounds and composition of present invention are particularly useful in ICE activity inhibition and thereby can be used as drug for treating of diseases mediated by IL-1, apoptosis, IGIF and IFN-γ, as well as inflammations, autoimmune diseases, bone-destructive disorder, infections, disorder associated with cell proliferation, degenerative and necrotic disorders. Uses of claimed compounds and compositions as well as methods for production of N-acylamino compounds also are disclosed.

EFFECT: effective interleikyn-1beta converting enzyme inhibitors.

64 cl, 35 ex, 35 tbl, 21 dwg

FIELD: medicine, gastroenterology.

SUBSTANCE: traditional eradication therapy should be supplemented with licopid at the dosage of 10 mg per os once daily before breakfast for 10 d. The present innovation prevents transfer of microorganisms into inactive form, accelerates restoration of mucosal epithelial layer in gastroduodenal area, provides complete eradication of microorganisms, that in its turn, favors to prevent disease exacerbation and restoration of gastroduodenal functions.

EFFECT: higher efficiency of therapy.

3 dwg, 2 ex

FIELD: biotechnology, biochemistry.

SUBSTANCE: invention relates to producing the biologically active complex eliciting antioxidant and immunomodulating activity and used in medicine, cosmetics, veterinary science and food industry. The biologically active complex preparing by enzymatic hydrolysis of muscle tissue represents complex of biologically active compounds involving carnosine and anserine in the amount 85-97 wt.-% of the native content of these components in poultry muscle tissue, 1-7 weight parts of amino acids, 0.5-12 weight parts of oligopeptides of molecular mass 10 kDa, not above, and 0.1-15 weight parts of cyclic and polycyclic phenolic compounds as measured for 1 weight part of carnosine and anserine in the complex. This complex is prepared by enzymatic hydrolysis of milled and homogenized water muscle tissue in preferable dilution homogenate with water in the range 0.2-0.6 and with using proteolytic enzymes in the amount 2-5 wt.-% of the protein content and working at pH 4.5-8.5 and at enhanced temperature being preferably at 45-65°C. Product is isolated as extract or powder prepared in drying the extract. Invention provides enhancing effectiveness of the claimed complex.

EFFECT: improved method for preparing, valuable properties of complex.

7 cl, 6 tbl, 6 ex

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