New anti-diabetic agents

FIELD: organic chemistry, medicine, endocrinology, pharmacy.

SUBSTANCE: invention relates to compounds of the formula (1) or its pharmaceutical salts wherein A means

R1 means hydrogen atom (H), (C1-C6)-alkyl (including branched alkyl and cycloalkyl), -(CH2)aNHW1, -(CH2)bCOW2, -(CH2)cOW3, -CH(Me)OW4, -(CH2)d-C6H4-W5, -(CH2)eSW6 wherein a = 2-5; b = 1-4; c = 1-2; d = 1-2; e = 1-3; W1 means -COW6, -CO2W6, -SO2W6; W2 means -OH, -NH2, -OW6, -NHW6; W3 means hydrogen atom (H), W6; W4 means H, W6; W5 means H, -OH, -OMe; W6 means (C1-C6)-alkyl, benzyl, optionally substituted phenyl wherein optional substitutes (up to two groups) are taken among (C1-C3)-alkyl, (C1-C3)-alkoxy-group, fluorine (F) and/or chloride (Cl) atoms; R2 means H, -(CH2)nNH-C5H3N-Y wherein n = 2-4; Y means H, F, Cl atoms, -NO2 and -CN; or R1 and R2 represent in common -(CH2)p- wherein p = 3 or 4; X is taken among: (i) one L-alpha-aminoacyl group taken among Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr and Val, or two such groups that represent Arg and Ile; (ii) groups -R3CO wherein R3 represents H, (C1-C6)-alkyl (including branched alkyl and cycloalkyl) or phenyl; (iii) groups -R4COOC, -(R5)(R6)OCO wherein R4 means H, (C1-C6)-alkyl (including branched alkyl and cycloalkyl), benzyl or optionally substituted phenyl wherein substituted (up to two groups) are taken among (C1-C3)-alkyl, (C1-C3)-alkoxy-group, F and Cl atoms; each R5 and R6 means independently H or (C1-C6)-alkyl; or R5 and R6 mean in common -(CH2)m- wherein m means a whole number 4-6; and (iv) methoxycarbonyl, ethoxycarbonyl and benzyloxycarbonyl groups; R7 is taken among pyridyl and optionally substituted phenyl wherein substitutes (up to two groups) are taken among (C1-C3)-alkyl, (C1-C3)-alkoxy-group, F, Cl atoms, -NO2, -CN and -CO2H; R8 means H or (C1-C3)-alkyl; R9 means H, (C1-C6)-alkyl, phenyl or (C1-C6)-alkoxy-group and under condition that indicated compound doesn't represent N(Z-Val)-2-cyanopyrrolidine. Compounds of the formula (1) are inhibitors of DP-IV and can be used in pharmaceutical compositions in treatment of the tolerance disturbances to glucose and diabetes mellitus of type 2.

EFFECT: valuable medicinal properties of compounds and compositions.

26 cl, 2 tbl, 19 ex

 

This invention relates to a series of new compounds, which are useful for the treatment of type 2 diabetes, impaired glucose tolerance and some other conditions.

Background of invention

Consider that the enzyme dipeptidyl peptidase IV (EC, here designated as DP-IV, also known as DPP-IV or DAP-IV) included in the regulation of the activities of certain hormones. One of these hormones is glucoheptonate peptide 1 (GLP-1), which is involved in the regulation of glucose levels in the blood after a meal and which under the action of DP-IV is translated from its active forms of GLP-1(5-36) in the inactive GLP-1(7-36). In cases of type 2 diabetes and impaired glucose tolerance when hyperglycemia can lead to tissue damage, it would be useful to enhance the effect of endogenous GLP-1. In this regard, inhibitors of DP-IV proposed as drug candidates for the treatment of type 2 diabetes and impaired glucose tolerance. For example, Demuth and other (WO 97/40832) disclosed the action of N-isolatin1encoding on the level of glucose in the blood on suitable animal models. However, this connection may not be strong enough to be a viable therapeutic agent. Stronger inhibitors of DP-IV are disclosed in Jenkins and others (WO 95/15309) and Villhauer (WO 98/19998), but they are prone to instability and cyclization in solution. This nestabil the awn would lead to difficulties in obtaining and storing the material of appropriate quality for therapeutic human use. Thus, there remains a need for an agent that inhibits DP-IV in vivo, but stable enough for commercial production.

Brief description of the invention

Found a series of derivatives that are chemically stable but undergo metabolic activation after administration to the subject, highlighting the highly active inhibitors of DP-IV. In this field such derivatives are usually referred to as prodrugs. Compounds of the present invention are useful for the treatment of type 2 diabetes and impaired glucose tolerance, as well as other States in which the enhanced activity of the hormone, usually inactivated by DP-IV, brings therapeutic benefit.

The compounds of this invention are derivatives of 1-(2'-aminoacyl)-2-cyanopyrrolidine General formula 1:

where a is selected from groups 2, 3 and 4; X is selected from aminoaniline group corresponding to one of the natural amino acids, acyl group, R3CO., the group R4COOC(R5)(R6)OCO, methoxycarbonyl, ethoxycarbonyl and benzyloxycarbonyl; R1selected from H, C1-C6alkyl residue, (CH2)andNHW1, (CH2)bCOW2, (CH2)cOW3CH(Me)OW4, (CH2)d-C6H4-W5, (CH2)eSW6where a is 2-5, b is 1-4, with equal 1-2, d RA is but 1-2, e is 1-3, W1means COW6, CO2W6or SO2W6, W2IT denotes, NH2, OW6or NHW6, W3denotes H, or W6, W4denotes H, or W6, W5denotes H, HE or OMe and W6represents C1-C6alkyl, optionally substituted phenyl, optionally substituted heteroaryl or benzyl, and R2selected from H and (CH2)n-C5H3N-Y, where n is 2-4 and Y denotes H, F, Cl, NO2or CN, or R1and R2together represent -(CH2)p-where p is 3 or 4; R3selected from H, C1-C6the alkyl and phenyl; R4selected from H, C1-C6of alkyl, benzyl, and optionally substituted phenyl; R5and R6each independently selected from N and C1-C6the alkyl or together represent -(CH2)m-where m is 4-6; R7selected from pyridyl and optionally substituted phenyl; R8selected from N and C1-C3the alkyl and R9selected from H, C1-C6of alkyl, C1-C6alkoxy and phenyl.

The present invention relates to new compounds defined above, pharmaceutical compositions in which at least one active agent is a compound of the present invention, the use of such compositions for the treatment of certain medical conditions and methods for the s treatment, when in need of the treatment of the subject is administered the compounds of this invention.

Detailed description of the invention

On the one hand, the present invention relates to a series of new compounds that are prodrugs of therapeutically useful inhibitors of DP-IV. Compounds of the present invention are derivatives of 1-(2'-aminoacyl)-2-cyanopyrrolidine following General formula 1

In this formula a is a group selected from 2, 3 and 4:

The dashed bond (dashed line) denotes a covalent bond, which connects the nitrogen atom of group a with 1.

The group X is acyl or oxycarbonyl group. Suitable groups:

(i) aminoaniline group corresponding to one of the natural amino acids: alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), Proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr) and valine (Val);

(ii) acyl group, R3CO., where R3selected from a hydrogen atom, a C1-C6alkyl groups and phenyl groups;

(iii) allocentrically group, R4COOC(R5)(R6)OCO, where R4 selected from a hydrogen atom, a C1-C6alkyl group, benzyl group or phenyl group which may have a substituent, C1-C3alkyl group, and R5and R6each independently is a hydrogen or C1-C6alkyl group, or R5and R6together represent polymethene chain -(CH2)m-where m is an integer of 4-6; and (iv) methoxycarbonyl, etoxycarbonyl and beneluxtunnel.

R1represents a side chain of naturally occurring amino acids or their analogues. More specifically, R1selected from a hydrogen atom, a C1-C6alkyl residues, (CH2)aNHW1, (CH2)bCOW7, (CH2)cOW3CH(Me)OW4, (CH2)d-C6H4-W5and (CH2)eSW6where a is 2-5, b is an integer of 1-4, with 1 or 2, d is 1 or 2, e is an integer of 1-3, W1means COW6, CO2W6or SO2W6, W2IT denotes, NH2, OW6or NHW6, W3denotes H, or W6, W4denotes H, or W6, W5denotes H, HE or OMe and W6represents C1-C6alkyl, optionally substituted phenyl, optionally substituted heteroaryl or benzyl. Suitable optional substituents heteroaryl and phenyl groups include C 1-C3alkyl and C1-C3alkoxygroup, and fluorine atoms and chlorine. May contain up to two substituents.

R2is a hydrogen atom or a group - (CH2)nNH-C5H3N-Y, where p is an integer of 2-4, C5H3N is a divalent pyridinium fragment and Y denotes a hydrogen atom, a halogen atom such as fluorine atom or chlorine, a nitro-group, a cyano.

Alternative R1and R2together may represent -(CH2)p-where p is 3 or 4.

R7selected from pyridyl and optionally substituted phenyl. Suitable optional substituents include C1-C3alkyl group, a C1-C3alkoxygroup, halogen atoms, the nitro-group, a cyano and carboxypropyl. May contain up to two substituents.

R8denotes a hydrogen atom or a C1-C3alkyl group.

R9denotes a hydrogen atom, a C1-C6alkyl group, a C1-C6alkoxygroup or phenyl group.

In the description of the present description, the term "alkyl" includes linear and branched alkyl groups, and cycloalkyl group. For example, C1-C6alkyl includes methyl, ethyl, ISO-propyl, tert-boutelou, neopentylene and tsiklogeksilnogo group. Also the term "heteroaryl" includes monocyclic five - and six-membered aromatic ring, which contain from one to three heteroatoms selected from nitrogen, oxygen and sulfur. For example, heteroaryl includes pirolli, pyridyl, furyl, thienyl, imidazolyl, thiazolyl, isoxazolyl, thiadiazolyl, pyrimidyl and pyrazinyl.

Some of the compounds of the present invention have acidic or basic properties and, thus, can exist in the form of salts. Up until such salts are non-toxic and pharmaceutically acceptable in other respects, they are included in the scope of this invention. Examples of such salts include, but are not limited to) acetate, hydrochloride, sulfate, phosphate and benzoate basic compounds and salts of sodium, potassium and tetraalkylammonium acidic compounds.

Except for the case when R1is H, compounds of General formula 1 have two stereogenic center (asymmetric carbon atoms), marked below With*. Preferably illustrated stereochemistry of these two provisions. Some variants of R1and X allow you to enter additional stereogenic centers, thus, the compounds of this invention can exist as epimere, including diastereomers. It is believed that all such optical isomers, including mixtures of such optical isomers are included in the scope of this invention.

In a preferred embodiment, the present invention includes a compound of General formula 1, in which R1different from N, and R2if present, denotes N. In a more preferred embodiment, R1represents C1-C6alkyl.

In another preferred embodiment, the present invention includes a compound of General formula 1 in which R1denotes H and a is selected from the groups corresponding to the General formulas 2 and 4, where R2represents - (CH2)nNH-C5H3N. In a more preferred embodiment, n is 2 and Y represents CN. In the most preferred embodiment, the NH group is in position 2 and group CN is in position 5 of the pyridine ring.

In another preferred embodiment, the present invention includes a compound of General formula 1, where And denotes a group of General formula 2 and X denotes aminoaniline group. In one more preferred embodiment X represents aminoacyl group, based on the primary amino acid such as lysine or arginine, most preferably arginine. In another more preferred embodiment X represents aminoaniline the group corresponding to glycine.

In another preferred embodiment, the present invention includes a compound of General formula 1, where And denotes a group of General formula 2 and X denotes a group R4COOC(R5)(R6)OCO. In one preferred embodiment, R4about the means C 1-C6alkyl. In another preferred embodiment, one of R5and R6denotes H and the other is methyl. Most preferably, when R4and one of R5and R6are metelli and the other of R5and R6is N.

In another preferred embodiment, the present invention includes a compound of General formula 1, where And denotes a group of General formula 2 and X denotes methoxycarbonyl group.

In another preferred embodiment, the present invention includes a compound of General formula 1, where And denotes a group of General formula 3.

In another preferred embodiment, the present invention includes a compound of General formula 1, where And denotes a group of General formula 4. In a more preferred embodiment, R8represents C1-C3alkyl and most preferably methyl. In another more preferred embodiment, R9represents C1-C3alkyl or C1-C3alkoxy, and most preferably methyl or methoxy.

Compounds that include distinctive characteristics of more than one of these preferred options are particularly preferred. The most preferable variant of the present invention is a compound selected from:

(2S)-1-((2'S)-2'-(1"-acetoacetanilide)-3,3'-dimethyl shall ethanol)pyrrolidin-2-carbonitrile;

(2S)-1-(N'-(1"-acetoxyacetyl)isoleucyl)pyrrolidin-2-carbonitrile;

(2S)-1-(N'-(methoxycarbonyl)isoleucyl)pyrrolidin-2-carbonitrile;

(2S)-1-((N')-(4"-oxopent-2"-EN-2"-yl)isoleucyl) pyrrolidin-2-carbonitrile;

(2S)-1-(pillsamaryl)pyrrolidin-2-carbonitrile;

(2S)-1-(orginalitati)pyrrolidin-2-carbonitrile;

(2S)-1-((2'S)-2'-(ecotoxicologically)-3,3'-dimethylbutanol)pyrrolidin-2-carbonitrile;

(2S)-1-((2'S)-2'-(1"-acetoacetanilide)-2'-cyclohexylethyl)pyrrolidin-2-carbonitrile;

(2S)-1-((2'S)-2'-(1"-acetoacetanilide)-4',4'-dimethylpentane)pyrrolidin-2-carbonitrile;

(2S)-1-(N'-(1"-acetoxyacetyl)-O'-tert-butylsilane)pyrrolidin-2-carbonitrile;

(2S)-1-(Nα-(1'-acetoxyacetyl)-Nωpair-toluensulfonyl)pyrrolidin-2-carbonitrile;

(2S)-1-(N'-(1'-acetoxyacetyl)-N-2"-(5"'-cyano-2"'-ylamino)ethyl)glycinyl)pyrrolidin-2-carbonitrile;

(2S)-1-(N'-(benzyloxycarbonyl)-O'-tert-butylthiophenol)-pyrrolidin-2-carbonitrile;

(2S)-1-(S'-tert-butyl-N'-(ethoxycarbonyl)cysteinyl)-pyrrolidin-2-carbonitrile;

(2S)-1-(Nω-acetyl-Nα-benzoylethyl) pyrrolidin-2-carbonitrile and

(2S)-1-(Nα-(acetyl)-Nω(benzyloxycarbonyl)ornithine)-pyrrolidin-2-carbonitrile.

Compounds of the present invention can be obtained by standard who tym techniques which are well known to experts in the field of organic chemistry. In many cases a suitable source substance is an amine of General formula 5, in which R1and R2have the same meanings as defined previously.

The synthesis of such compounds is described, for example, in Jenkins and others (WO 95/15309), Villhauer (WO 98/19998), Ashworth and others (Bioorg. Med. Chem. Lett., 1996, 6(10), 1163-66) and Li and others (Arch. Biochem. Biophys. 1995, 323(1), 148-54). Compounds not described in detail in these publications can be obtained by the usual modification shown in these ways. The stage included in the formation of compounds of this invention from compounds of General formula 5, depend on the nature of group A.

X = aminoaniline group,

Scheme 1

Scheme 1 illustrates receiving data connection in two stages. Chn denotes the side chain of amino acids. Depending on the amino acid Chn can be H (glycine), CH3(alanine), (CH3)2SN (valine), (CH3)2SNSN2(leucine), CH3CH2CH(CH3) (isoleucine), C6H5CH2(phenylalanine), NOSE6H4CH2(tyrosine)8H6NCH2(tryptophan), NOESN2(aspartic acid), HOOCCH2CH2(glutamic acid), H2NOCCH2(asparagine), 2NOCCH2CH2(glutamine), NON2(serine), CH3CH(OH) (threonine), HSCH2(cysteine), CH3SCH2CH2(methionine), C3H3N2CH2(histidine), H2N(CH2)4(lysine) and H2NC(:NH)(CH2)3(arginine). As will be clear to experts in the practical chemistry of peptides, some of these side chains contain functional groups that are reactive under the conditions necessary to implement the condensation of two fragments. Data functional groups must be protected by suitable masking groups. Such groups are described, for example, in the work "Protective Groups in Organic Synthesis, by T.W. Greene, Wiley-Interscience, 1981. Chn* denotes the same side chains, but with the necessary protective groups.

Also PG denotes a protective group for aminophenol.

1-(2'-Aminoacyl)-2-cyanopyrrolidine 5 condensing protected with appropriate amino acid 6 with the intermediate product 7, using a variety of conditions that are well known in the field of peptide chemistry. Typically, the two components are dissolved in a suitable solvent, which is typically an aprotic solvent, such as dichloromethane, dimethylformamide or a mixture, and the solution is cooled to 0°With or below. The solution is to add one or two equivalents of amine bases of the tion, such as diisopropylethylamine or dimethylaminopyridine. Then add the condensing agent and the mixture is stirred until consumption of the original substances, as shown, for example, analytical thin-layer chromatography. If the reaction proceeds slowly, then speed up the process, it may be appropriate to give the mixture to warm to room temperature. Suitable condensing agents include DCC (dicyclohexylcarbodiimide), THIEF (hexaphosphate benzotriazol-1 yloxy)Tris(dimethylamino)phosphonium), Rumor® ((hexaphosphate benzotriazol-1 yloxy)triprolidine), PyBroP® (hexaphosphate patrimonialization) and HBTU (hexaphosphate O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyl-Urania).

Remove protection from the intermediate product 7 gives the target compound 1A(namely, the compound of General formula 1 with a group of General formula 2 and the amino acid residue X).

Instead of 6, you can use protected appropriately Proline, getting the same connection, where X is propilovyi balance. All protected amino acids are commercial products.

Scheme 2

Scheme 2 illustrates receiving data connections. The original substance 5 is treated with acylchlorides or anhydride in an aprotic solvent and in PR is the presence of amine base, such as described above, when receiving the product 1B. If R3=H, it is impossible to use acylchlorides or anhydride. In this case, use a mixed anhydride. The reagent is conveniently obtained from formic acid and acetic anhydride.

Scheme 3

Scheme 3 illustrates receiving data connections. The original substance 5 is treated with para-nitrophenylarsonic 8 in an aprotic solvent and in the presence of an amine base, such as described previously, getting the product 1c. Carbonate receive according to the method described in Alexander and others, J. Med. Chem. 31, 318, 1988.

X = methoxycarbonyl, etoxycarbonyl, benzyloxycarbonyl.

Scheme 4

Scheme 4 illustrates receiving data connections. The original substance 5 is treated with chloroformate in an aprotic solvent and in the presence of an amine base, such as described previously, getting the product 1D. As benzylchloride (BnOCOCl) is not very stable, it is convenient to replace benzyl 1-succinimidylester (BnOCONSu). This and all chloroformiate are commercial products.

Scheme 5And

Scheme 5 illustrates receiving data connections. the similar substance 5 A(namely, the compound of General formula 5 with R2=H) is subjected to interaction with the aldehyde 9 in the presence of an acid catalyst, such as, for example, para-toluensulfonate acid. The interaction is carried out in a solvent such as cyclohexane or toluene, at elevated temperatures such as the boiling point of the solvent. Constantly remove the water or azeotropic distillation, or by using a desiccant, such as activated molecular sieve.

The aldehyde 9 is a commercial product.

Scheme 6

Scheme 6 illustrates receiving data connections. Starting material 5 is subjected to interaction with 1,3-dicarbonyl compound 10 in an aprotic solvent and in the presence of an amine base as described previously, at room temperature.

Dicarbonyl compounds 10 are commercial products or can be well-known methods.

Of course, other methods of synthesis. They usually differs from the above described procedure stages. Figure 7 shows two examples.

Scheme 7

The intermediate product 11 receives the above-described methods. It is possible to condense with prolinamide 12 by methods described in scheme 1, receiving sub the full product 13. It can be degidratiruth processing triperoxonane anhydride, obtaining the target compound. Alternative intermediate product 11 can condense with preminition 14, directly receiving the target connection.

Compounds of General formula 1 (compound of the present invention) undergo metabolism in the body, giving compounds of General formula 5. These metabolites are inhibitors of DP-IV.

Scheme 8

As discussed earlier, believe that the inhibitors of DP-IV are useful for the treatment of some medical conditions. Therefore, the compounds of the present invention are useful for the treatment of such conditions. In particular, the compounds of the present invention are useful for the treatment of impaired glucose tolerance or type 2 diabetes. They can also be useful in the treatment of reproductive disorders such as infertility due to polycystic ovary syndrome. Another application is the treatment of insufficiency of growth hormone, leading to low growth. The scope of this invention also covers other medical condition.

For use in the treatment of these disorders, the compounds of the present invention typically include pharmaceutical compositions and prepare appropriately for the selected route of administration. Such compositions are : the m aspect of the present invention. The pharmaceutical composition may include other pharmaceutically acceptable excipients, in General known in the field, such as agents that adds volume, diluents, dispersing agents, preservatives, colorants, flavoring agents and the like. The choice of fillers depends on the method of preparation and the introduction of the composition. This composition you can enter the usual well-known in this field means. For example, the composition can be prepared in the form of tablets, capsules, syrup or powder for oral administration in the form of tablets or wafers for sublingual or transbukkalno injection, suppository for rectal or vaginal application, solution or powder for nasal administration, cream or lotion for topical application, the plaster for transdermal application, or in the form of a solution or suspension for subcutaneous, intramuscular or intravenous injection. Input injection form may include encapsulated and others known in the field forms with controlled release of active substances that are suitable for the introduction of the depot. The preferred composition is a tablet for oral administration.

On the third hand, the invention includes a method of treating glucose intolerance or type 2 diabetes, wherein the patient in need of such treatment, in which W ill result therapeutically effective amount of the above compounds. The regimen dose is usually determined by the attending physician, taking into account the specific features of the patient. The usual dose is from 1 to 500 mg once a day or up to four times per day.

The previous description is additionally illustrated below by several examples. They are intended to illustrate implementation of the present invention, but in no way limit described herein scope of the invention.

EXAMPLES

Solvents and reagents are usually used as received without additional purification. Structures of all intermediates were confirmed by the method of1H NMR. The final products, in addition, is characterized by a method of mass spectrometry and/or elemental analysis.

Example 1-(2S)-1-((2'S)-2'-(1"-acetoxyethylamine)-3,3'-dimethylbutanol)pyrrolidin-2-carbonitrile

A solution of (2S)-1-((2'S)-2'-amino-3,3'-dimethylbutanol)-pyrrolidin-2-carbonitrile hydrochloride (180 mg, 0.73 mmol; obtained according to Jenkins and others, WO 95/15309), α-acetoxyethyl para-nitrophenylphosphate (220 mg, 0.82 mmol; obtained according to Alexander and others, J. Med. Chem. 31, 318, 1988) and triethylamine (90 mg, 0.90 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NHCO 3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 3:7)to give white solid, identified as specified in the title compound (170 mg, 0.50 mmol, 68%).

MS: ionization when elektrorazpredelenie {M+H}+=340,2.

1H NMR (CDCl3): δ 1,02, of 1.03 (N, HS), 1,42 of 1.46 (3H, m), 2,03, was 2.05 (3H, 2×C)2,15 was 2.25 (4H, m), 3,69 is 3.76 (2H, m), 4,23-to 4.28 (1H, m), 4,77-rate 4.79 (1H, m), 5,43 (1H, d, J=9.5 Hz), 6.73 x-6,77 (1H, m) h/million

Example 2 - (2S)-1-(N'-(1"-Acetoxyacetyl)isoleucyl)-pyrrolidin-2-carbonitrile

A solution of (2S)-1-(isoleucyl)pyrrolidin-2-carbonitrile hydrochloride (500 mg, 2.04 mmol; obtained according to Jenkins and others, WO 95/15309), α-acetoxyethyl para-nitrophenylphosphate (610 mg, of 2.27 mmol; obtained according to Alexander and others, J. Med. Chem. 31, 318, 1988) and triethylamine (250 mg, of 2.50 mmol) in dichloromethane (40 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent:EtOAc:petrify 60-80°, 3:7)to give a colourless oil identified as specified in the title compound(480 mg, 1.48 mmol, 70%).

MS: ionization when elektrorazpredelenie {M+N}+=340,0.

1H NMR (CDCl3): δ 0,86-0,89 (6N, m)of 0.92 to 0.97 (1H, m), 1.41 to a 1.45 (3H, m), 150-1,80 (2H, m), 2,02 (3H, d, J=5,2 Hz), 2,14-of 2.27 (4H, m), 3,60 of 3.75 (2H, m), to 4.23-4.26 deaths (1H, t, J=7,6 Hz), of 4.77 (1H, d, J=2.3 Hz), and 5.30-of 5.50 (1H, m), 6.73 x-6,77 (1H, m) h/million

Example 3 - (2S)-1-(N'-(Methoxycarbonyl)isoleucyl) pyrrolidin-2-carbonitrile

A solution of (2S)-1-(isoleucyl)pyrrolidin-2-carbonitrile hydrochloride (300 mg, 1,22 mmol; obtained according to Jenkins and others, WO 95/15309), methylcarbamate (125 mg, 1.3 mmol) and triethylamine (150 mg, 1.50 mmol) in dichloromethane (40 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 4:6)to give a colourless oil identified as specified in the title compound (310 mg, of 1.16 mmol, 95%).

MS: ionization when elektrorazpredelenie {M+H}+=268,2.

1H NMR (CDCl3): δ 0,85-0,95 (6N, m), 1,10-1,25 (1H, m), and 1.54-1.77 in (2H, m), 2,11-of 2.26 (4H, m), 3,62 (3H, s), 3,66-with 3.79 (2H, m), is 4.21 (1H, t, J=9,2 Hz), 4,74-4,78 (1H, m), and 5.30 (1H, d, J=9.1 Hz) h/million

Example 4 - (2S)-1-((N')-(4"-Oxopent-2"-EN-2"-yl)isoleucyl)-pyrrolidin-2-carbonitrile

A solution of (2S)-1-(isoleucyl)pyrrolidin-2-carbonitrile hydrochloride (150 mg, 0.61 mmol; obtained according to Jenkins and others, WO 95/15309), 2,4-pentanedione (68 mg, of 0.68 mmol) and triethylamine (75 mg, 0.75 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 7:3)to give a colourless oil identified as specified in the title compound (85 mg, 0.29 mmol, 47%).

MS: ionization when elektrorazpredelenie {M+N}+=292,3.

1H NMR (CDCl3): δ 0,87-0,98 (6N, m), 1,19-1,25 (1H, m), 1,61 was 1.69 (2H, m)of 1.84 (3H, s)to 1.98 (3H, s), 2,15 was 2.25 (4H, m), 3,49-of 3.54 (1H, m), 3,62 at 3.69 (1H, m), 3.95 to 3,98 (1H, m), 4.75 V-rate 4.79 (1H, m), to 4.98 (1H, s), 11,09 (1H, d, J=8.1 Hz) h/million

Example 5 - (2S)-1-(Pillsamaryl)pyrrolidin-2-chronical

(a) (2S)-1-(Isoleucyl)pyrrolidin-2-carbonitrile

To a solution of hemihydrate BOC-isoleucine (0.96 g, 4 mmol) and Rover® (2,34 g, 4.5 mmol) in dichloromethane (25 ml) is added DIPEA (of 1.74 ml, 10 mmol). To this solution was added solid (S)-pyrrolidin-2-carbonitrile hydrochloride (0,60 g, 4.5 mmol), and then another portion of DIPEA (697 μl, 4 mmol). Reactionary see what camping is stirred for 2 hours. The solvent is removed on a rotary evaporator and transfer the residue in ethyl acetate. The resulting solution was washed with 0.3 M sodium bisulfate (2x), saturated sodium bicarbonate (2x), water and saturated sodium chloride. The organic phase is dried with anhydrous sodium sulfate and removing the solvent on a rotary evaporator. The residue is dissolved in a mixture of TFA (95%) and water (5%). After 1 hour TFA and water are removed under reduced pressure and the residue is triturated with ether, the resulting precipitate. This precipitate is collected and dried in vacuum, obtaining trifenatate salt specified in the header of the product as a white solid; yield of 0.58 g (1.8 mmol, 45%).

(b) (2S)-1-(Pillsamaryl)pyrrolidin-2-carbonitrile

To a solution of BOC-glycine (0.21 g, 1.2 mmol) and Rover® (of 0.62 g, 1.2 mmol) in dichloromethane (3 ml) is added DIPEA (522 μl, 3 mmol). To this solution was added the product of example 5A (0.28 g, 0.9 mmol), and then another portion of DIPEA (157 μl, 0.9 mmol). The reaction mixture was stirred over night. The solvent is removed on a rotary evaporator and transfer the residue in ethyl acetate. The resulting solution was washed with 0.3 M sodium bisulfate (2x), saturated sodium bicarbonate (2x), water and saturated sodium chloride. The organic phase is dried with anhydrous sodium sulfate and removing the solvent on a rotary evaporator. The residue is dissolved in a mixture of TFA (95%) and water (5%) and paramesh the live mix throughout the night. A large part of TFA and water are removed under reduced pressure. The residue is purified HPLC with reversed phase, receiving trifenatate salt of the final product as a white powder; yield 171 mg (50%).

Example 6 - (2S)-1-(Orginalitati)pyrrolidin-2-chronical

To a solution of Boc-Arg(Mtr)-OH (of 0.58 g, 1.2 mmol) and Rover® (of 0.62 g, 1.2 mmol) in dichloromethane (3 ml) is added DIPEA (522 μl, 3 mmol). To this solution was added the product of example 5A (0.28 g, 0.9 mmol), and then another portion of DIPEA (157 μl, 0.9 mmol). The reaction mixture was stirred over night. The solvent is removed on a rotary evaporator and transfer the residue in ethyl acetate. The resulting solution was washed with 0.3 M sodium bisulfate (2x), saturated sodium bicarbonate (2x), water and saturated sodium chloride. The organic phase is dried with anhydrous sodium sulfate and removing the solvent on a rotary evaporator. The residue is dissolved in a mixture of TFA (95%) and water (5%) and the mixture is stirred over night. A large part of TFA and water are removed under reduced pressure and the residue triturated with ether. The ether layer is decanted and the residue purified by the method of HPLC with reversed phase, receiving trifenatate salt of the final product as a white powder; yield 83 mg (19%).

Example 7 - (2S)-1-((2'S)-2'-(Ecotoxicologically)3,3'-dimethylbutanol)pyrrolidin-2-carbonitrile

A solution of (2S)-1-((2'S)-2'-amino-3,3'-dimethylbutanol)-pyrrolidin-2-carbonitrile hydrochloride (150 mg, 0.61 mmol; obtained according to Jenkins and others, WO 95/15309), acetoxymethyl para-nitrophenylphosphate (168 mg, 0.66 mmol; obtained according to Alexander and others, J. Med. Chem. 31, 318, 1988) and triethylamine (70 mg, 0.70 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 4:6)to give white solid, identified as specified in the title compound (188 mg, of 0.58 mmol, 95%).

MS: ionization when elektrorazpredelenie {M+H}+=to 326.1.

1H NMR (CDCl3): δ 1,03 (N, C)of 2.09 (3H, s), 2,16-of 2.24 (4H, m), 3.72 points-of 3.77 (2H, m), 4,25 (1H, d, J=9.6 Hz), 4,77-4,80 (1H, m), of 5.68 (1H, d), of 5.68 (2H, s) h/million

Example 8 - (2S)-1-((2'S)-2'-(1"-Acetoacetanilide)2'-cyclohexylethyl)pyrrolidin-2-carbonitrile

A solution of (2S)-1-((2'S)-2'-amino-2'-cyclohexylethyl)-pyrrolidin-2-carbonitrile of triptoreline (100 mg, 0.28 mmol; obtained according to Jenkins and others, WO 95/15309), α-acetoxyethyl para-nitrophenylphosphate (76 mg, 0.29 mmol; obtained according to Alexander and others, J. Med. Che. 31, 318, 1988) and triethylamine (35 mg, 0.35 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 4:6)to give white solid, identified as specified in the title compound (43 mg, 0.12 mmol, 41%).

MS: ionization when elektrorazpredelenie {M+H}+=366,2.

1H NMR (CDCl3): δ 0,97-to 1.21 (4H, m), of 1.40 to 1.48 (3H, m), 1,67-to 1.77 (7H, m), 2,02 (3H, d, J=7.8 Hz), 2,11-of 2.26 (4H, m), 3,65-to 3.73 (2H, m), 4,16-4,22 (1H, m), was 4.76 (1H, d, J=4, 2 Hz), are 5.36-5,41 (1H, m), 6.73 x-6,77 (1H, m,) h/million

Example 9 - (2S)-1-((2'S)-2'-(1"-Acetoacetanilide)4',4'-dimethylpentane)pyrrolidin-2-carbonitrile

A solution of (2S)-1-((2'S)-2'-amino-4',4'-dimethylpentyl)-pyrrolidin-2-carbonitrile of triptoreline (100 mg, 0.30 mmol; obtained according to Jenkins and others, WO 95/15309), α-acetoxyethyl para-nitrophenylphosphate (87 mg, 0.33 mmol; obtained according to Alexander and others, J. Med. Chem. 31, 318, 1988) and triethylamine (40 mg, 0.40 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate(70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 4:6)to give white solid, identified as specified in the title compound (32 mg, 0.09 mmol, 31%).

MS: ionization when elektrorazpredelenie {M+H}+=354,2.

1H NMR (CDCl3): δ to 0.97, and 0.98 (N, HS), 1,41 was 1.43 (3H, m), 1,44-of 1.62 (2H, m), 2,03 (3H, d, J=2.3 Hz), 2,16-of 2.21 (4H, m), 3,61-3,63 (1H, m), 3,74-of 3.78 (1H, m), 4,45-to 4.52 (1H, m), 4.75 V-of 4.77 (1H, m), 5,24-of 5.29 (1H, m), 6.73 x-is 6.78 (1H, m) h/million

Example 10 - (2S)-1-(N'-(1"-Acetoxyacetyl)-O'-tert-butylsilane)pyrrolidin-2-carbonitrile

A solution of (2S)-1-(O'-tert-butylsilane)pyrrolidin-2-carbonitrile hydrochloride (30 mg, 0.11 mmol; obtained according to Jenkins and others, WO 95/15309), α-acetoxyethyl para-nitrophenylphosphate (32 mg, 0.12 mmol; obtained according to Alexander and others, J. Med. Chem. 31, 318, 1988) and triethylamine (20 mg, 0.20 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80° With, 4:6)to give white solid, identified as specified in the title compound (14 mg, of 0.038 mmol, 35%).

MS: ionization when elektrorazpredelenie {M+N}+=370,1.

1H NMR (CDCl3): δ 1,11-1,15 (N, m), 1.41 to a 1.45 (3H, m), 2,04 (3H, d, J=4.9 Hz), 2,10-of 2.15 (2H, m), 3,43-3,62 (5H, m), 3,90-4,00 (1H, m), 4,50 with 4.65 (1H, m), to 4.73 (1H, d, J=5,2 Hz), the 5.45 5,72 (1H, m), 6,76-6,79 (1H, m,) h/million

Example 11 - (2S)-1-(Nα-(1'-Acetoxyacetyl)-Nωpair-toluensulfonyl)pyrrolidin-2-carbonitrile

A solution of (2S)-1-(Nωpair-toluensulfonyl)-pyrrolidin-2-carbonitrile of triptoreline (100 mg, 0.20 mmol; obtained according to Jenkins and others, WO 95/15309), α-acetoxyethyl para-nitrophenylphosphate (61 mg, 0.23 mmol; obtained according to Alexander and others, J. Med. Chem. 31, 318, 1988) and triethylamine (30 mg, 0.30 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 7:3)to give white solid, identified as specified in the title compound (51 mg, 0.10 mmol, 49%).

MS: ionization when elektrorazpredelenie {M+is} +=509,0.

1H NMR (CDCl3): δ of 1.41 to 1.48 (6N, m)and 1.51-1.69 in (2H, m), is 2.05 (3H, d, J=18.3 Hz), 2,12-of 2.28 (5H, m)to 2.41 (3H, s), 2,86-of 2.93 (2H, m), 3,63-of 3.64 (2H, m), of 4.38 was 4.42 (1H, m), 4,72-to 4.73 (1H, m,), 4,74-4,79, 5,10-5,20 ( 1H, 2xm), 5,54-5,62 (1H, m), 6,74-6,79 (1H, m), 7,29 (2H, d, J=7,7 Hz), 7,71 (2H, d, J=8,4 Hz) h/million

Example 12 - (2S)-1-(N'-(1'-Acetoxysilane)-N-(2"-(5"'-cyanopyridine-2"'ylamino) ethyl) glycine)pyrrolidin-2-carbonitrile

A solution of 1-([2-[(5-cyano-2-yl)amino]ethyl]amino]-acetyl)-2-cyano-(S)-pyrrolidin bis(triptoreline) (100 mg, 0,19 mmol; obtained according Villhauer and others, WO 98/19998), α-acetoxyethyl para-nitrophenylphosphate (56 mg, 0.21 mmol; obtained according to Alexander and others, J. Med. Chem. 31, 318, 1988) and triethylamine (50 mg, 0.50 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4. saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°S, 9:1)to give white solid, identified as specified in the title compound (13 mg, 0.03 mmol, 16%).

MS: ionization when elektrorazpredelenie {M+H}+=429,2.

1H NMR (CDCl3): δ 1,21-of 1.32 (3H, m), 1,40-of 1.46 (1H, m), 1,99-of 2.05 (4H, m), 2,17-2,31 (4H, m), 3,50-3,63 (6N, m), 4,40-4,50 (1H, is), of 4.77 (1H, d, J=5,9 Hz), 6,45-of 6.49 (1H, m), 6,68-6,77 (1H, m), 7,44-of 7.48 (1H, m), 8,32 (1H, s) h/million

Example 13 - (2S)-1-(N'-(Benzyloxycarbonyl)-O'-tert-butylthiophenol) pyrrolidin-2-carbonitrile

A solution of (2S)-1-(O'-tert-butylthiophenol)pyrrolidin-2-carbonitrile hydrochloride (35 mg, 0.12 mmol; obtained according to Jenkins and others, WO 95/15309), benzylchloride (32 mg, 0.13 mmol) and triethylamine (24 mg, 0.24 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: chloroform:methanol, 98:2)to give white solid, identified as specified in the title compound (47 mg, 0.12 mmol, 100%).

MS: ionization when elektrorazpredelenie {M+H}+=388,3.

1H NMR (CDCl3): δ 1,10-1,30 (3H, m), 1,18 (N, C)2,00 at 2.45 (4H, m), 3,55-3,70 (1H, m), 3,85-4,00 (2H, m), 4,30-and 4.40 (1H, m), 4,70-4,80 (1H, m), 5,07 (2H, s), of 5.75 (1H, d, J=8,15 Hz), 7,20 was 7.45 (5H, m) h/million

Example 14 - (2S)-1-(S'-tert-butyl-N'-(ethoxycarbonyl)-cysteinyl)pyrrolidin-2-carbonitrile

A solution of (2S)-1-(S')-tert-butylstannyl)pyrrolidin-2-carbonitrile of triptoreline (100 mg, 0.27 mmol; obtained according to the about Jenkins and others, WO 95/15309), ethylchloride (35 mg, 0.32 mmol) and triethylamine (50 mg, 0.50 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: EtOAc:petrify 60-80°, 8:2)to give white solid, identified as specified in the title compound (30 mg, 0,092 mmol, 35%).

MS: ionization when elektrorazpredelenie {M+H}+=328,1.

1H NMR (CDCl3): δ of 1.18 (3H, t, J=7 Hz), 1,30 (N, C), 2,17-of 2.24 (4H, SIRM), 2,82-to 2.85 (2H, m), 3,70-3,82 (2H, SIRM), of 4.05-4.09 to (2H, m), 4,48-a 4.53 (1H, m), 4,74-of 4.77 (1H, m), 5,41-5,44 (1H, m) h/million

Example 15 - (2S)-1-(Nω-Acetyl-Nα-6eos)pyrrolidin-2-carbonitrile

A solution of (2S)-1-(Nω-acetylethyl)pyrrolidin-2-carbonitrile of triptoreline (100 mg, 0.22 mmol; obtained according to Jenkins and others, WO 95/15309), benzoyl chloride (343 mg, 0.24 mmol) and triethylamine (45 mg, 0.45 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO32SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: chloroform:methanol, 97:3)to give white solid, identified as specified in the title compound (83 mg, 0.22 mmol, 100%).

MS: ionization when elektrorazpredelenie {M+N}+=387,6.

1H NMR (CDCl3): δ 1,56-of 1.78 (4H, SIRM), was 1.94 (3H, s), 2,12-of 2.20 (4H, SIRM), 3,21 is 3.23 (2H, m), 3,59-and 3.72 (2H, m), 4,65-4,69 (2H, m), 5,07 (2H, s), 5,18-to 5.21 (1H, m), 6,69-6,72 (1H, m), 7.24 to 7,34 (5H, m) h/million

Example 16 - (2S)-1-(Nα-(Acetyl)-Nω(beneluxtunnel)-ornithine)pyrrolidin-2-carbonitrile

A solution of (2S)-1-Nω(benzyloxycarbonyl)ornithine)-pyrrolidin-2-carbonitrile of triptoreline (100 mg, 0.23 mmol; obtained according to Jenkins and others, WO 95/15309), acetylchloride (20 mg, 0.26 mmol) and triethylamine (50 mg, 0.50 mmol) in dichloromethane (25 ml) was stirred at room temperature for 18 hours. After that, the solvent is removed in vacuum and transfer the residue in ethyl acetate (70 ml). This solution was washed with 0.3 M KHSO4saturated NaHCO3, water and saturated salt solution, dried (Na2SO4) and evaporated. The residue is purified by way of flash chromatography (eluent: chloroform:methanol, 97:3)to give white solid, identified as specified in the title compound (49 mg, 0.13 mmol, 5%).

MS: ionization when elektrorazpredelenie {M+N}+=371,2.

1H NMR (CDCl3): δ 1,30-of 1.65 (4H, m), 1,75-of 1.95 (2H, m), 1,90 (3H, s), 2,10-to 2.40 (4H, m), 3,10-3,30 (2H, m), 3,65-are 3.90 (2H, m), 4,70-of 4.90 (2H, m), 5,90-6,00 (1H, m), 7,30 is 7.50 (4H, m), 7,70-7,80 (2H, m) h/million

Example 17 In vitro inhibitory activity relative to DP-IV

Compounds of the previous examples were investigated as inhibitors of DP-IV in the ways described in the work of Ashworth and others (Bioorg. Med. Chem. Lett. 1996, 6(10), 1163-66). Not found to have significant inhibitory activity up to 10 μm, it indicates that the prodrugs of this invention are at least 1000 times less active than the active inhibitors from which they derive. Therefore, we can conclude that any visible in vivo activity due to bioconversion in the original inhibitors.

Example 18 In vivo activity on the model of glucose tolerance

The activity of these compounds investigated in male rats Zucker Fatty Rats 10 - and 20 weeks of age. Animals do not feed during the night and then injected them orally through a feeding tube investigational compound (10 mg/kg) in the form of a solution. After an hour take the blood samples (200 µl) from the tail vein for determination of the baseline (t=0) of glucose, then the animals give oral glucose (1 g/kg in the form of a 40% wt./about. solution). Then take blood samples at t=10, 20, 30, 60 and 120 min Glucose determine enzymatic experiments is observed. Typical results are presented in the table below.

The reference compound in the above experiments is the compound of example 11 of WO 95/15309. It is the reference compound, derivatives of which are prodrugs of examples 2-6 of the present description.

From the above results it is clear that the prodrug effective to reduce hyperglycemia after entering glucose, but they are not always as effective as a comparative compound in the early time points. This is what you would expect for prodrugs that are converted in high yield to the original drug. Results at early time points are the result of the fact that the necessary metabolic conversion of circulating prodrugs.

In a separate experiment investigated the connection given at the same dose (10 mg/kg), but 12 hours before oral input of glucose. The results are shown below.

ConnectionBlood glucose (mg/DL); AVG. Val.±srcmatch., n-4
t=0t=10t=20t=30t=60t=120AUC
Filler84,2±3,7145,5±6,6134,3±8,0127,2�B1; the 10.1122,9±8,7to 112.2±8,64556±458
Example 183,7±3,8113,5±10,8111,3±9,991,9±11,899,3±10,0116,9±to 14.42430±591

AUC (area under the curve "concentration - time") is strongly reduced, indicating that the prodrug has a significant antihyperglycemic activity continued for 12 hours.

The above results show that the compounds of the present invention demonstrate antihyperglycemics activity after oral administration in suitable animal models of glucose intolerance. Therefore, it is expected that they will be effective in the treatment of impaired glucose tolerance and type 2 diabetes in humans. In addition, the results of in vivo confirm that prodrugs into active inhibitors of DP-IV in circulation and that they can be used in the treatment of all other pathologies for which such inhibitors are assumed as therapeutic agents.

Example 19 - Pharmaceuticals

19A - Tablets 50 mg

Tablets containing 50 mg of the compound of example 1, is obtained from the following ingredients:

Connection example 154,5 g
Corn starch53,5 g
Hydroxypropylcellulose13.5 g
Carboxymethylcellulose calcium11,0 g
Magnesium stearate2.0 g
Lactose165,5 g
Only400,0 g

Substances are mixed and then pressed, receiving 200 tablets 200 mg, each containing 50 mg of the compound of example 1.

The compounds of examples 2, 3 and 5 are prepared separately in the form of tablets in a similar way. The compounds of examples 4 and 6-16 similarly separately formed into tablets containing 100 mg of the corresponding connection.

19C - Suppositories, 100 mg

Suppositories containing 100 mg of the compound of example 2 is obtained from the following ingredients:

The compound of example 2154,5 g
Corn starch210.0 g
Colloidal silicon dioxide2.5 g
Povidone 3049,0 g
Magnesium stearate23,0 g
Adipic acid57,0 g
Sodium bicarbonate43,0 g
Sodium lauryl sulfate5.0 g
456,0 g
Only1000,0 g

Substances are mixed and then pressed, receiving suppositories 1 g, each of which contains 100 mg of the compound of Example 2. The compounds of examples 1, 3-5 and 6-16 are prepared separately in the form of suppositories in the same way.

1. The compound of General formula 1 or its pharmaceutically acceptable salt

where And denotes

R1selected from H, C1-C6of alkyl (including branched alkyl and cycloalkyl), (CH2)aNHW1, (CH2)bCOW2, (CH2)cOW3CH(Me)OW4, (CH2)d-C6H4-W5and (CH2)eSW6where a is 2-5, b is 1-4, with equal 1-2, d equals 1-2, e equals 1-3, W1means COW6, CO2W6or SO2W6, W2IT denotes, NH2, OW6or NHW6, W3denotes H, or W6, W4denotes H, or W6, W5denotes H, HE or OMe and W6represents C1-C6alkyl, benzyl, optionally substituted phenyl, where the optional substituents (up to two groups selected from C1-C3of alkyl, C1-C3alkoxy, F and/or Cl;

R2selected from H or (CH2)nN-C 5H3N-Y, where n is 2-4 and Y is selected from H, F, Cl, NO2and CN, or R1and R2together represent -(CH2)p-where p is 3 or 4;

X is chosen from:

(i) one L-alpha-aminoacyl group selected from Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tight and Val, or two such groups, which are Arg and Ile;

(ii) R3CO., where R3denotes H, C1-C6, alkyl (including branched alkyl and cycloalkyl) or phenyl,

(iii) groups, R4COOC(R5)(R6)OCO, where R4denotes H, C1-C6alkyl (including branched alkyl and cycloalkyl), benzyl or optionally substituted phenyl in which the substituents (up to two groups selected from C1-C3of alkyl, C1-C3alkoxy, F or Cl, and R5and R6each independently represents N or C1-C6, alkyl, or R5and R6together represent -(CH2)m-where m is an integer of 4-6, and

(iv) methoxycarbonyl, ethoxycarbonyl and benzyloxycarbonyl groups;

R7selected from pyridyl and optionally substituted phenyl, where the substituents (up to two groups selected from C1-C3of alkyl, C1-C3alkoxy, F, Cl, NO2CN and CO2N;

R8denotes N or C1-C3alkyl and

R9denotes H, C1 -C6, alkyl, phenyl or C1-C6alkoxygroup,

provided that the compound is not N-(Z-Val)-2-cyanopyrrolidine.

2. The compound according to claim 1, where R1different from N and R2denotes N.

3. The compound according to claim 2, where R1represents C1-C6alkyl.

4. The compound according to claim 1, where R1denotes N, And denotes a group of General formula 2 or 4 and R2represents -(CH2)nNH-C5H3N-Y.

5. The compound according to claim 4, where n is 2 and Y represents CN.

6. The compound according to claim 5, where the Deputy NH is in position 2 and cyano is in position 5 of the pyridine ring.

7. The compound according to claim 1, where a denotes a group of General formula 2 and X denotes aminoaniline group.

8. The connection according to claim 7, where aminoaniline group is leilei or orgininally group.

9. The connection of claim 8, where aminoaniline group is orgininally group.

10. The connection according to claim 7, where aminoaniline group is picaninny group.

11. The compound according to claim 1, where a denotes a group of General formula 2 and X denotes a group R4COOC(R5)(R6)OCO.

12. Connection to item 11, where R4represents C1-C6alkyl.

13. Connection claim 11, where one of R5and R6denotes H and the other is methyl.

14. Connection to item 11, where R4means m is Teal, one of R5and R6denotes H and the other is methyl.

15. The compound according to claim 1, where a denotes a group of General formula 2 and X denotes methoxycarbonyl.

16. The compound according to claim 1, where a denotes a group of General formula 4.

17. Connection P16, where R8represents C1-C3alkyl.

18. The connection 17, where R8denotes methyl.

19. Connection P16, where R9stands With1-C3alkyl or C1-C3alkoxy.

20. The connection according to claim 19, where R9denotes a methyl or methoxy group.

21. The compound according to claim 1, chosen from:

(2S)-1-((2'S)-2'-(1"-acetoacetanilide)-3,3'-dimethylbutanol)pyrrolidin-2-carbonitrile,

(2S)-1-(N'-(1"-acetoxyacetyl)isoleucyl)-pyrrolidin-2-carbonitrile,

(2S)-1-(N'-(methoxycarbonyl)isoleucyl)pyrrolidin-2-carbonitrile,

(2S)-1-((N')-(4"-oxopent-2"-EN-2"-yl)isoleucyl)-pyrrolidin-2-carbonitrile,

(2S)-1-(pillsamaryl)pyrrolidin-2-carbonitrile,

(2S)-1-(orginalitati)pyrrolidin-2-carbonitrile,

(2S)-1-((2'S)-2'-(ecotoxicologically)-3,3'-dimethylbutanol)pyrrolidin-2-carbonitrile,

(2S)-1-((2'S)-2'-(1"-acetoacetanilide)-2'-cyclohexylethyl)pyrrolidin-2-carbonitrile,

(2S)-1-((2'S)-2'-(1"-acetoacetanilide)-4',4'-dimethylpentane)pyrrolidin-2-carbonitrile,

(2S)-1-(N'-(1"-and maximoserver)-O'-tert-butylsilane)pyrrolidin-2-carbonitrile,

(2S)-1(Nα-(1'-acetoxyacetyl)-Nωpair-toluensulfonyl)pyrrolidin-2-carbonitrile,

(2S)-1-(N'-(1'-acetoxyacetyl)-N-(2"-(5"'-cyanopyridine-2"'-ylamino)ethyl)glycinyl)pyrrolidin-2-carbonitrile,

(2S)-1 -(N'-(benzyloxycarbonyl)-O'-tert-butylthiophenol)-pyrrolidin-2-carbonitrile,

(2S)-1-(S'-tert-butyl-N'-(ethoxycarbonyl)cysteinyl)-pyrrolidin-2-carbonitrile,

(2S)-1(Nω-acetyl-Nα-benzoylethyl)pyrrolidin-2-carbonitrile and

(2S)-1-(Nα-(acetyl)-Nω(benzyloxycarbonyl)ornithine)-pyrrolidin-2-carbonitrile.

22. The compound according to any one of claims 1 to 21, selected from the optical isomers of the compounds according to any one of the preceding paragraphs.

23. The compound according to any one of claims 1 to 21, selected from the pharmaceutically acceptable salts of the compounds according to any one of the preceding paragraphs.

24. Pharmaceutical composition for the treatment of impaired glucose tolerance or type 2 diabetes containing agent selected from compounds of General formula 1 or its pharmaceutically acceptable salt

where And denotes

R1selected from H, C1-C6of alkyl (including branched alkyl and cycloalkyl), (CH2)aNHW1, (CH2)bCOW2, (CHsub> 2)cOW3CH(Me)OW4, (CH2)d-C6H4-W5and (CH2)eSW6where a is 2-5, b is 1-4, with equal 1-2, d equals 1-2, e equals 1-3, W1means COW6, CO2W6or SO2W6, W2IT denotes, NH2, OW6or NHW6, W3denotes H, or W6, W4denotes H, or W6, W5denotes H, HE or OMe and W6represents C1-C6alkyl, benzyl, optionally substituted phenyl, where the optional substituents (up to two groups selected from C1-C3of alkyl, C1-C3alkoxygroup, F and/or Cl;

R2selected from H or (CH2)nNH-C5H3N-Y, where n is 2-4 and Y is selected from H, F, Cl, NO2and CN, or R1and R2together represent -(CH2)p-where p is 3 or 4;

X is chosen from:

(i) one L-alpha-aminoacyl group selected from Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tight and Val, or two such groups, represents Arg and Ile;

(ii) R3CO., where R3denotes H, C1-C6, alkyl (including branched alkyl and cycloalkyl) or phenyl;

(iii) groups, R4COOC(R5)(R6)OCO, where R4denotes H, C1-C6alkyl (including branched alkyl and cycloalkyl), benzyl or optional someseni is phenyl, in which the substituents (up to two groups selected from C1-C3of alkyl, C1-C3alkoxy, F or Cl, and R5and R6each independently represents N or C1-C6, alkyl, or R5and R6together represent -(CH2)m-where m is an integer of 4-6, and

(iv) methoxycarbonyl, ethoxycarbonyl and benzyloxycarbonyl groups;

R7selected from pyridyl and optionally substituted phenyl, where the substituents (up to two groups selected from C1-C3of alkyl, C1-C3alkoxy, F, Cl, NO2CN and CO2N;

R8denotes N or C1-C3alkyl and

R9denotes H, C1-C6, alkyl, phenyl or C1-C6alkoxygroup.

25. The compound or compounds according to any one of claims 1 to 23 as the active agent to obtain drugs for the treatment of impaired glucose tolerance or type 2 diabetes.

26. A method of treating impaired glucose tolerance or type 2 diabetes, which includes the introduction in need of such treatment to the patient a therapeutically effective amount of the compounds (compounds) according to any one of claims 1 to 23.



 

Same patents:

The invention relates to a new method of obtaining derivatives of 3-pyrrolin-2-carboxylic acid of the formula I, where1- C1-C6-allyloxycarbonyl; R2is hydroxyl, WITH1-C4-alkoxy or their ammonium salts, the removal of base sulfoxylate residue from the compounds of formula II, where R1and R2as above; R3- C1-C6-alkyl, benzyl, trifluoromethyl, naphthyl, phenyl which may be substituted by a residue comprising SN3, NO2, halogen

The invention relates to new and useful acaricidal and insecticidal arilpirolii compounds, methods for their preparation, intermediate compounds for their production, methods of producing these compounds, and methods of control of ticks and insects

The invention relates to new derivatives of 2-methyl-3-etoxycarbonyl-5-(2'-cyano-3'-aminopropyl-2'-enylidene)pyrrolin - 2-it-4 or 2-(2'-cyano-3'-aminopropyl-2'-enylidene)indolinone-3 of the General formula 1

< / BR>
where, if R = COOC2H5, R1= CH3, R2+ R3= (CH2)5(a) R2+ R3= CH2CH2OCH2CH2(b) R2= H, R3= CH2C6H5(C) R2= H, R3= CH2C6H4OCH3-4 (g) R2= H, R3= CH2C6H3(OCH3)-3,4 (d) R + R1= CH = CH-CH = CH R2= R3= H (e) R2= H, R3= (CH2)6(W) R2= H, R3= CH2CH2N(C2H5)2(C) R2= H, R3= CH2CH(OH)CH2OH and R2+ R3= CH2CH2OCH2CH2(K) R2+ R3= CH2CH2N(CH3)CH2CH2(l) R2+R3=CH2CH2= H, R3= CH2C6H4OCH3-4 (o) R2= H, R3= CH2C6H3(OCH3)-3,4 (p) R2= H, R3= CH(CH3)C6H5(p) R2= H, R3= CH2CH2C6H5(c) R2= H, R3= CH2CH2C6H3(OCH3)-3,4 (t) R2= H, R3= CH(CH3)CH2C6H5(u) R2= H, R3= C6H5(f) R2= H, R3= C6H4OCH3-4 (x) with antihypertensive activity

FIELD: organic chemistry, pharmaceutical compositions.

SUBSTANCE: invention relates to new (N-substitutes glycyl)-2-cyanopyrrolidines of formula I , wherein R is adamantly, substituted in 3- and/or 5-site with one or more substituents, selected from group including C1-C10-alkyl, OR1 (R1 is C1-C10-alkyl, C1-C8-alkanoyl, -CO-NR4R5, wherein R4 and R5 are independently from one another hydrogen, cyclohexyl, C1-C10-alkyl, phenyl optionally substituted with C1-C10-alkyl or C1-C10-alkoxy), in free form or in form of acid additive salt. Claimed compounds inhibit dipeptidyl-peptidase IV (DPP-IV) activity and useful in pharmaceutical composition for treatment of conditions mediated by DPP-IV, such as insulin-independent diabetes mellitus and obesity.

EFFECT: new pharmaceutical compounds inhibiting dipeptidyl-peptidase IV.

5 cl, 1 dwg, 4 tbl, 12 ex

The invention relates to new derivatives of nitrogen-containing heterocyclic compounds of the formula

or their pharmaceutically acceptable salts, where R1represents H, COCOR2, COOR3or SO2R3, R2is1-6alkyl, C1-6alkenyl,5-7cycloalkyl, 2-thienyl, 3-thienyl, phenyl or substituted phenyl, R3is phenylalkyl,represents a saturated five-membered nitrogen-containing heterocyclic ring with one nitrogen atom or benzododecinium saturated six-membered nitrogen-containing heterocyclic ring;is oxazol, oxadiazole or thiazole, And is associated with carbon atom of the five-membered heteroaromatic rings and represents COO(CH2)mAr,where R1has the values listed above or is CONR4(CH2)mAr or (CH2)mO(CH2)nAr and R1cannot be COCOR2or SO2R3, R4represents H or<

The invention relates to a derivative of D-Proline General formula

< / BR>
or

< / BR>
where R is SH, benzyl or phenyl, optionally substituted by a hydroxy-group or a lower alkoxygroup, or a group of the formula

< / BR>
R1is hydrogen or halogen; X represents -(CH2)n-; -CH(R2)(CH2)n-; -CH2O(CH2)n-; CH2NH-; benzyl, -C(R2)=CH-; CH2CH (OH)- or thiazol-2,5-diyl; Y represents-S -; (CH2)n; -O-; -NH-; -N (R2)-; -CH=CH-; -NHC(O)NH-; -N(R2)C(O)N(R2)-; -N[CH2WITH6H3(OCH3)2]-; -N(CH2WITH6H5)-; -N(CH2WITH6H5)C(O)N(CH2WITH6H5)-; -N(alkoxyalkyl)-; -N(cyclooctylmethyl)-; 2,6-pyridyl; 2,5-furanyl; 2,5-thienyl; 1,2-cyclohexyl; 1,3-cyclohexyl; 1,4-cyclohexyl; 1,2-naphthyl; 1,4-naphthyl; 1,5-naphthyl; 1,6-naphthyl or diphenylene; 1,2-phenylene; 1,3-phenylene or 1,4-phenylene, where phenylenebis group optionally substituted by 1-4 substituents selected from the group comprising halogen, lower alkyl, lower alkoxygroup, the hydroxy-group, carboxypropyl, -COO-lower thiazolyl, 2-oxo[1,2,3,5] oxadiazolyl, 5-thioxo[1,2,4]oxadiazolyl and 5-tert-butylsulfonyl[1,2,4] oxadiazolyl; X' represents -(CH2)n-; (CH2)nCH(R2)-; -(CH2)nOCH2-; -NHCH2-; benzyl, -CH= C(R2)-; -CH(OH)CH2or thiazol-2,5-diyl; R2denotes lower alkyl, lower alkoxygroup or benzyl and n = 0-3, their pharmaceutically acceptable salts, mono - and diesters, except (R)-1-[(R)- and (R)-1-[(S)-3-mercapto-2-methylpropionyl] pyrrolidin-2-carboxylic acid; medicinal product with amyloidoses activity, and the method of obtaining these derivatives

The invention relates to nitrate ACE-inhibitor of formula I or II, where Y is phenyl, X is C(RIIIRIV, RIII, RIV, RVand RVI- hydrogen containing stoichiometric amount of nitric acid

The invention relates to a new derived neurotrophic N-glyoxal-propyl ether of the formula 1

< / BR>
where R1means1-C5alkyl linear or branched chain, possibly substituted C3-C6cycloalkyl,3- WITH5- WITH6-cycloalkyl, or AG1where Ar1selected from the group consisting of 2-tanila, 2-furanyl, 2-thiazolyl or phenyl; X is oxygen; Y represents oxygen or NR2where R2is hydrogen; Z means WITH2-C6alkyl or alkenyl with a linear or branched chain, which is substituted by one or more than one position AG2or3-C6cycloalkyl, where AG2selected from the group consisting of phenyl, 2-, 3 - or 4-pyridyl, phenyl, substituted methylenedioxy, and phenyl, having one to three substituents, which independently represent a chlorine or1-C4alkoxy, or pharmaceutically acceptable salts or hydrates

The invention relates to N-(N'-substituted glycyl)-2-cyanopyrrolidine formula I, where R denotes: a)1R1aN (CH2)m-, where R1means pyridinoline or pyrimidinyl fragment, optional one - or disubstituted independently of one another by halogen, trifluoromethyl, cyano - or nitro-group; R1adenotes hydrogen or C1-C8alkyl, m is equal to 2,3, b)3-C12cycloalkyl, optional one-deputizing in position 1 WITH1-C3hydroxyalkyl,) R2(CH2)n- where either R2denotes phenyl, optional one-, two - or tizamidine selected independently of each1-C4alkoxygroup, halogen or phenylthiourea, optional one-deputizing in the phenyl ring with hydroxymethyl; or denotes a C1-C8alkyl, [3.1.1] bicyclic carbocyclic fragment, optional single or mnogozalny1-C8the alkyl, pyridinoline or nattily fragment, or cyclohexenyl, or substituted and n is 1-3, or R2denotes fenoxaprop; and n is 2; d) (R3)2CH(CH2)2-, where each R3independently represents phenyl; d) R4(CH2)p-, where R4ebony in position 1 WITH1-C3hydroxyalkyl, W) R5that means indanyl piperidinyl fragment, optionally substituted benzyl, and [2.2.1] or [3.1.1] bicyclic carbocyclic fragment, optional single or mnogozalny1-C8by alkyl, substituted or1-C8alkyl, optionally one or mnogozalny independently from each other hydroxy-group, hydroxymethyl or phenyl, optional one - or disubstituted independently selected from each other WITH1-C4the alkyl, C1-C4alkoxygroup or halogen, in free form or in the form of an acid additive salt

The invention relates to new derivatives carbapenem formula I, where R1and R2may be the same or different, and each represents a modifiable group that can be hydrolyzed in the body, selected from 1-alkanoyloxy, 1-alkoxycarbonylmethyl, 5-methyl-1,3-dioxolan-2-he-4-ylmethyl; R3and R4may be the same or different and each represents lower alkyl, or R3and R4together with the adjacent nitrogen atom form a cyclic amino; or pharmaceutically acceptable salts

The invention relates to new derivatives of 1-methylcarbamate General formula (I) described in the claims

FIELD: medicine.

SUBSTANCE: application of hypoglycemic agent as nateglinide for treating affected glucose metabolism for the purpose to prevent or deteriorate the onset of pronounced non-insulin dependent diabetes mellitus or prevent or deteriorate the onset of other manifestations of affected glucose metabolism. The innovation proves nateglinide's capacity to restore early phase of insulin secretion, that is it should be applied as an intensifier of insulin secretion.

EFFECT: higher efficiency of application.

7 cl, 5 ex, 1 tbl

FIELD: organic chemistry, medicine, hormones.

SUBSTANCE: invention describes imidazole derivatives of the formula (I) , racemic-diastereomeric mixtures and optical isomers, pharmaceutical salts wherein ---- represents an optional bond; R1 represents hydrogen atom (H), -(CH2)m-C(O)-(CH2)m-Z1, -(CH2)m-Z1; R2 represents hydrogen atom (H), or R1 and R2 are joined with nitrogen atoms to which they are bound forming compounds represented by formulae (Ia), (Ib) or (Ic) wherein R3 represents -(CH2)m-E-(CH2)m-Z2; R4 represents hydrogen atom (H) or -(CH2)m-A1; R5 represents (C1-C12)-alkyl, (C0-C6)-alkyl-C(O)-NH-(CH2)m-Z3 and optionally substituted phenyl; R6 represents hydrogen atom (H); R7 represents (C1-C12)-alkyl or -(CH2)m-Z4; m = 0 or a whole number from 1 to 6; n is a whole number from 1 to 5. Proposed compounds bind with subtypes of somatostatin receptors selectively.

EFFECT: valuable properties of compounds.

20 cl, 13776 ex

FIELD: medicine, endocrinology.

SUBSTANCE: one should introduce mildronate per 0.25 g thrice daily for internally about 30 min before meals and additionally - neuromultivit per 1 tablet twice daily internally after meals. Treatment should be carried out at the background of insulinotherapy. Therapy course lasts for 8 wk. The innovation suggested provides stable improvement of functional state of peripheral nervous system due to complex impact upon pathogenesis of polyneuropathy in this category of patients.

EFFECT: higher efficiency of therapy.

1 cl, 1 ex, 1 tbl

FIELD: medicine, ophthalmology.

SUBSTANCE: one should introduce berlition parabulbarly per 0.5 ml for 10 d followed by the intake of its tableted form per 300 mg once daily for 1 mo. Additionally, since the 1st d of therapy one should introduce memoplant per 1 tablet thrice daily for 6 wk. The innovation leads to either complete or partial resolution of hemorrhages, decreases the quantity of exudates and microaneurysms, enables to narrow the areas of retinal ischemia that, in its turn, improves patient's acuity.

EFFECT: higher efficiency of therapy.

2 ex, 4 tbl

FIELD: organic chemistry, biochemistry, medicine.

SUBSTANCE: invention proposes compounds inhibiting activity of enzyme SGLT-2 of the formula (I)

wherein R1, R2 and R2a mean independently hydrogen atom, hydroxyl (OH), OR5, alkyl, CF3, -OCHF2, -OCF3, CR5i or halogen atom, or two radicals among R1, R2 and R2a in common with adjacent carbon atoms can form annelated 5-membered heterocycle; R3 and R4 mean independently hydrogen atom, OH, OR5, O-aryl, OCH2-aryl, alkyl, cycloalkyl, CF3, -OCHF2, -OCF3, halogen atom, -CN, -CO2R5b, - CO2H, -CONR6R6a, -NHCOR5c, -NHSO2R5d, -NHSO2-aryl, aryl, -SR5c, -SOR5f, -SO2R5g, -SO2-aryl, or 5-, 6- or 7-membered carbocycle or heterocycle; R5, R5a, R5b, R5c, R5d, R5e, R5f, RR5g, R5h and R5i means independently alkyl; R6, R6a, R6b, R6c and R6d mean independently hydrogen atom, alkyl, aryl, alkylaryl or cycloalkyl, or R6 and R6a in common with adjacent nitrogen atoms form annelated 5-, 6- or 7-membered heterocycle; A means oxygen (O), sulfur (S) atom, NH or (CH2)n wherein n = 0-3. Also, invention proposes a method for treatment of diabetes mellitus and related diseases by using the above described compound in doses inhibiting activity of SGLT-2 taken as a single substance or in combination with other therapeutic agents.

EFFECT: valuable medicinal and biochemical properties of inhibitors.

19 cl, 2 tbl, 82 ex

FIELD: organic chemistry, vitamins, medicine, pharmacy.

SUBSTANCE: invention relates to a new compound of the formula (I): wherein X means hydrogen atom or hydroxy group; R1 and R2 that can be similar or different mean hydrogen atom, (C1-C4)-alkyl; R3 means hydrogen atom, methyl group, fluorine or chlorine atom. Also, invention relates to its esters able to hydrolysis in vivo in combination with pharmaceutically acceptable acids. Also, invention relates to a pharmaceutical composition eliciting the inhibitory activity with respect to proliferation and promoting differentiation of cells and comprising the effective dose of compound of the formula (I) in common with pharmaceutically acceptable carriers and/or excipients. Also, invention relates to applying compound of the formula (I) for preparing a medicine used in treatment and prophylaxis of disease characterizing by abnormal differentiation of cells and/or proliferation of cells.

EFFECT: valuable medicinal properties of compounds.

13 cl, 3 sch, 3 tbl, 6 ex

FIELD: chemical-pharmaceutical industry, pharmacy.

SUBSTANCE: method involves preparing human insulin ester by transpeptidation reaction of porcine insulin in the excess of threonine di-tert.-butyl ester in an aqueous-organic medium in the presence of trypsin, inhibition of reaction by acidification, purification of prepared insulin ester by chromatography method, removal of protecting groups with trifluoroacetic acid and purification of prepared human crude insulin. Preparing part of the combined preparation of short effect by dissolving the prepared insulin with concentrations 15%, 25% or 50% in diluted acid and mixing with preserving agent solutions, isotonic agent and substances with buffer capacity. Preparing part of the combined preparation of prolonged effect by dissolving remaining insulin in diluted acid also followed by addition of zinc ions acid solutions and protamine sulfate, mixing with buffered solutions of m-cresol, phenol and glycerol, keeping the solution up to formation of crystals and combination of prepared parts of the combined insulin preparation. The transpeptidation reaction is carried out in the weight ratio trypsin to porcine insulin = 1:(300-1000) and before acidification of reaction medium additional inhibition of reaction is carried out by dilution of the reaction mixture with water by 2-3 times. Insulin ester is purified by HPLC method followed by precipitation of fractions with ester derivative in the presence of zinc ions, removal of protecting groups and preparing crystals of crude insulin that are purified by repeated HPLC method. Both purifying processes are carried out by using sorbent DIASOGEL ODS (C18) as immobile phase with particles size from 15 mcm and pores size 120 , and at the first stage 0.06 M glycine-HCl buffer with 0.015 M of ammonium sulfate and propanol-2 with concentration from 20% to 35% at pH 2.5 is used as mobile phase, and at the second stage 0.05 M acetate buffer with the content of propanol-2 from 10% to 20% at pH 2.5 is used. In preparing both parts of the combined preparation crystals of insulin are dissolved in diluted acid by stage wherein firstly finely dispersed suspension of insulin crystals in water is prepared followed by addition of diluted acid to it. In preparing the combined preparation of prolonged effect a mixture obtained after mixing with buffered solutions of m-cresol, phenol and glycerol is kept at temperature 18-21°C for 20-22 h. Invention provides the development of effective, economy method for preparing the combined preparation of human insulin with low immunological properties that allows enhancing yield of insulin in the process of its manufacturing.

EFFECT: improved preparing method.

13 ex

FIELD: medicine, endocrinology, chemical-pharmaceutical industry, hormones.

SUBSTANCE: method involves preparing human insulin ester by transpeptidation reaction of porcine insulin in the molar excess of threonine di-tert.-butyl ester in an aqueous-organic medium in the presence of trypsin, inhibition of reaction by acidification, purification of preparing human insulin ester by chromatography, removal of protecting groups with trifluoroacetic acid and purification of prepared human crude insulin, its following dissolving in diluted acid and mixing with preserving agent solutions, isotonic agent and substances with buffer capacity. Preparing human insulin ester is carried out in the weight ratio trypsin to porcine insulin = 1:(300-1000) followed by additional inhibition of reaction by dilution of the reaction mixture with water by 2-3 times before acidifying. Purification of human insulin ester is carried out by HPLC method followed by precipitation of ester derivative fractions in the presence of zinc ions, removal of protecting groups and preparing human crude insulin crystals that is purified by repeated carrying out HPLC method. Both processes of purification are carried out by using sorbent DIASOGEL ODS (C18) as immobile phase with particles size from 15 mcm and pores size from 100 to 150 . At the first stage 0.06 M glycine - HCl buffer containing 0.015 M of ammonium sulfate and propanol-2 in the concentration from 20% to 35%, pH 2.5 is used as a mobile phase, and 0.05 M acetate buffer with the content of propanol-2 from 15% to 25%, pH 2.5 is used at the second stage. The purified human insulin is dissolved in diluted acid by stages wherein finely dispersed suspension of insulin crystals in water is prepared followed by addition a diluted acid to it. Invention provides the development of effective, economy method for preparing the ready medicinal formulation of insulin with short effect and with low immunological properties that provides reducing loss of insulin in the process of its preparing. Invention can be used in manufacturing ready insulin formulations of high quality and with short effect.

EFFECT: improved preparing method.

6 ex

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