Inhibitors of serine proteases

 

(57) Abstract:

Describes new inhibitors of serine proteases having piperidine side chain of the General formula (I), where (A) - H, optionally substituted D, L--hydroxyacetic, R1, R1-O-C(O)-, R1-SO2-, R2OOC-(CHR2)m- where R1selected from (1-12C)alkyl and (3-8C)cycloalkyl, and these groups can be substituted for IT, and from (7-15C)aralkyl, and each group R2independently represents H or has the same values that are specified for R1; m = 1, 2, or 3; In - bond, the amino acid of formula-NH-CH[CH2)pC(O)OH]-C(O)-, where R = 0, 1, 2 or 3; -N((1-12C)alkyl)-CH2-CO-D-1-Piq, or represents a D-amino acid having a hydrophobic side chain, or a and b taken together represent the residue R3R4N-CHR5-C(O)-, where R3and R4independently - R1, R1-O-C(O)-, R1-C(O) R1-SO2-, and R5is a hydrophobic or basic side chain; X Is L-amino acid with a hydrophobic side chain; Y IS-CO-NH-(1-6C)alkylene - C6H5, -R6where R6Is h or (1-6C)alkyl, -CONR7R8independently represent H or (1-6C)alkyl, or R7and R8taken together, represent (3-6C)alkylene, l is or its pharmaceutically acceptable salt. The above compounds have antikoaguliruyuschim action and can be used for the treatment or prevention of diseases mediated by the action of thrombin. Also describes a pharmaceutical composition containing this inhibitor. 2 C. and 13 C.p. f-crystals, 2 PL.

The invention relates to an inhibitor of serine proteases having piperidine side chain; to pharmaceutical compositions containing this inhibitor; and using the indicated inhibitor for the manufacture of a medicine for the treatment and prevention of diseases associated with the action of thrombin.

Serine proteases are enzymes that, in addition to its other functions, play an important role in the cascade of reactions of blood coagulation. Members of this group of proteases are, for example, thrombin, trypsin, factor VIIa, IXa, XA, XIa, XIIa and protein S.

Thrombin is a serine protease that regulates last stage of the cascade reactions of blood coagulation. The main function of thrombin is the degradation of fibrinogen with the production of fibrin monomers, which, by way of cross-links form an insoluble gel. In addition, thrombin regulation is dealt with by cascade reactions. It is also important to note that the thrombin functions at the cellular level, where it acts on specific receptors, causing, thereby, platelet aggregation, activation of endothelial cells and proliferation of fibroblast. Thus, thrombin plays a Central role of the regulator when the hemostasis and clot formation. Because thrombin inhibitors may have a wide range of therapeutic applications in this area was conducted extensive research.

In the development of synthetic inhibitors of serine proteases, and more specifically, thrombin increased interest in small synthetic peptides that are recognized by proteolytic enzymes by the mechanism similar to the recognition of natural substrates. In result, they have obtained new peptideprophet inhibitors, such as inhibitors of thrombin short-term actions.

Research on the development of more effective and selective inhibitors of thrombin tirelessly to continue to obtain inhibitors of thrombin, which could be introduced in small doses and that would have fewer side effects and less severe side effects. In addition, special attention is paid to primenimost Auda clinically effective in acute conditions, requiring treatment of diseases associated with the action of thrombin. However, to prevent diseases associated with the action of thrombin, such as myocardial infarction, thrombosis and shock requires prolonged therapy preferably by oral administration of certain doses of anticoagulant.

Most peptidebinding inhibitors of thrombin, as described in previous publications contain side chains of arginine. The problem is that these argininemia thrombin inhibitors have a low bioavailability when administered orally. The literature describes a number of inhibitors of thrombin, which instead contain arginine lysine side chains. It has been suggested that the lower basicity of lysine than arginine can lead to increased oral bioavailability of these inhibitors. Examples leinstermen of thrombin inhibitors are N-Me-Cha-Pro-Lys-COOH and their derivatives, described in Jones et al., J. Enzyme Inhibition, 9 (1995), 43-60, and inhibitors of N-Me-D - Phe-Pro-Lys-X, where X represents carboxamide or carboxylic acid, described by Lewis et al. Thrombosis and Haemostasis, 74(4) (1995), 1107-12. However, the expected improvement was observed, and oral Biotest few latinamerica and Haemostasis, 74(4) (1995), 1107-12). Other thrombin inhibitors have been described in WO 94/25051, where the lysine or arginine side chains were replaced aminocyclohexanone radicals, which may be regarded as isostere lysine.

The authors of this application it was found that inhibitors of serine proteases, especially thrombin inhibitors, XA and VIIa with piperidine side chain, of the formula I:

< / BR>
where A represents H, optionally substituted D,L--hydroxyacetic, R1, R1-O-C(O)-, R1-C(O)-, R1-SO2-, R2OOC- (CHR2)m-SO2, R2OOC-(CHR2)m-, H2NCO-(CHR2)m- or N-protective group, where R1selected from (1-12C)alkyl, (2-12C)alkenyl, (2-12C)quinil and (3-8C)cycloalkyl, and these groups can be optionally substituted (3-8C)cycloalkyl, (1-6C)alkoxy, oxo, HE, COOH, CF3or halogen, and from (6-14C)aryl, (7 - 15C)aralkyl and (8-16C)aralkyl; and aryl groups can be optionally substituted (1-6C)alkyl, (3-8C)cycloalkyl, (1 - 6C)alkoxy, HE, COOH, CF3or halogen; and each R2independently represents H or has the same values as above for R1; m=l, 2, or 3;

B is a bond, an amino acid of formula-NH-CH[(canil)-CH2-CO-, -N((2-12C)quinil)- CH2-CO-, -N(benzyl)-CH2-CO-D-1-Tiq, D-3-Tiq, D-Atc, Aic, D-1-Piq, D-3-Piq or L - or D-amino acid having a hydrophobic, basic or neutral side chain, where the specified amino acid may be optionally substituted N-(1-6C) alkyl; or a and b taken together represent the residue R3R4N-CHR5-C(O), where R3and R4independently represent R1, R1-O-C(O)-, R1-C(O)-, R1-SO2-, R2OOC-(CHR2)m-SO2, R2OOC -(CHR2)m-, H2NCO-(CHR2)m- or N-protective group, or one of R3and R4associated with R5forms a 5 - or 6-membered ring together with the group "N-C", with which it is associated, where the specified ring may be condensed with an aliphatic or aromatic 6-membered ring; R5is a hydrophobic, basic or neutral side chain;

X represents L-amino acid with a hydrophobic side chain, serine, threonine, a cyclic amino acid optionally containing an additional heteroatom selected from N, O or S, and optionally substituted (1-6C)alkyl, (1-6C)alkoxy, benzyloxy or oxo, or X is-NR2-CH2-C(O) -- or the fragment

< / BR>
or

< / BR>
where n=2, 3 P>6where R6represents H or (1-6C)alkyl, -CONR7R8and R7and R8independently represent H or (1-6C)alkyl, or R7and R8taken together, represent (3-6C)alkylene; or Y is a heterocycle selected from 2-thiazole, 2-thiazoline, 2-benzothiazole, 2-oxazole, 2-oxazoline and 2-benzoxazole where these heterocycles may be optionally substituted (1-6C)alkyl, phenyl, (1-6C)alkoxy, benzyloxy or oxo; and r=0, 1, 2 or 3;

or their prodrugs or pharmaceutically acceptable salts are strong and selective inhibitors. In addition, some compounds of the present invention have good bioavailability when administered orally.

Compounds of the present invention can be used for the treatment and prevention of thrombin-mediated and associated with the action of thrombin diseases. Such diseases include a number of thrombotic and prothrombotic conditions under which activates a cascade of reactions of blood coagulation and which include, but are not limited to, thrombosis of deep veins, pulmonary embolism, thrombosis, occlusion of the artery as a result of thrombosis or embolism, re-occlusion of the artery in the process or PEFC is ocedur, postoperative thrombosis or embolism veins, acute or chronic atherosclerosis, shock, myocardial infarction, cancer and cancer and neurodegenerative diseases. Compounds of the present invention can also be used as anticoagulants in the artificial circulation required for dialysis and surgery. Compounds of the present invention can also be used as in vitro anti -.

Preferred compounds of the present invention have the formula I, where X is an L-amino acid with a hydrophobic chain, serine, threonine, or-NR2-CH2-C(O). Other preferred compounds of formula I are compounds in which a is as defined above; B represents a bond, an amino acid of formula-NH-CH[(CH2)pC(O)OH]-C(O)- or its ester derivative, where p=0, 1, 2 or 3; -N((1-6C)alkyl)-CH2-CO-N((2-6C)alkenyl)-CH2-CO -, - N(benzyl)-CH2-CO-D-1-Tiq, D-3-Tiq, D-Atc, Aic, D-1-Piq, D-3-Piq or a D-amino acid having a hydrophobic side chain, where the specified amino acid may be optionally substituted N-(1-6C) alkyl; or a and b taken together represent the residue R3R4N-CHR5-C(O), and X represents tsiklicheskogo (1-6C) alkyl, (1-6C)alkoxy, benzyloxy or oxo; or X is-NR2-CH2-C(O) -- or the fragment

< / BR>
or

< / BR>
More preferred are the compounds of formula I, where a represents H, 2-hydroxy-3-cyclohexylpropionic-, 9-hydroxy-fluoren-9-carboxyl, R1, R1-SO2-, R2OOC-(CHR2)m-SO2, R2OOC-(CHR2)m-, H2NCO-(CHR2)m- or N-protective group, where R1selected from (1-12C)alkyl, (2-12C)alkenyl, (6-14C)aryl, (7-15C)aralkyl and (8-16C)aralkyl; each R2independently represents H or has the same values that were specified for R1B represents a bond, D-1-Tiq, D-3-Tiq, D-Atc, Aic, D-1-Piq, D-3-Piq or a D-amino acid having a hydrophobic side chain, where the specified amino acid may be optionally substituted N-(1-6C) alkyl; or A and B taken together represent the residue R3R4N-CHR5-C(O)-; Y is-CO-NH-(1-6C)alkylene-C6H5, - COOR6, -CONR7R8; or Y is a heterocycle selected from 2-thiazole, 2-thiazoline, 2-benzothiazole, 2-oxazole, 2-oxazoline and 2-benzoxazole.

Especially preferred are compounds in which a represents H, R1, R1-SO2or R2OOC-(CHR23R4N-CHR5-C(O), where at least one of R3and R4is R2OOC-(CHR2)mor R1-SO2- and the other independently is (1-12C) alkyl, (2-12C)alkenyl, (2-12C)quinil, (3-8C)cycloalkyl, (7-15C)aralkyl, R1-SO2or R2OOC-(CHR2)m-, a R5represents a hydrophobic side chain; Y represents-CO-NH-(1-6C)alkylene-C6H5, -COOR6where R6represents H or (1-3C)alkyl, -CONR7R8where R7and R8independently represent H or (1-3C-alkyl, or R7and R8taken together, represent a (3-5C)alkylene, or Y is a heterocycle selected from 2-thiazole, 2-benzothiazole, 2-oxazole or 2 - benzoxazole.

If a is R2OOC-(CHR2)m-, B preferably represents a D-amino acid having a hydrophobic side chain; or a and b taken together represent the residue R3R4N-CHR5-C(O)-, where at least one of R3and R4is R2OOC-(CHR2)m- and the other independently is (1-12C)alkyl, (2-6C)alkenyl, (3-8C)cycloalkyl, benzyl, R1-SO2or R2OOC-(CHR2)m- ; X is 2, -octahydrocyclopenta acid or [N(3-8C)cycloalkyl]-CH2-C(O)-. More preferred are compounds where a is HOOC-CH2-; B is D-Phe, D-Cha, D-Coa, D-Dpa, p-Cl-D-Phe, p-O methyl-D-Phe, p-O-ethyl-D-Phe, D - Nle, m-Cl-D-Phe, 3,4-di-OMe-D-Phe, D-Chg; or A and B taken together represent the residue R3R4N-CHR5-C(O)-, where at least one of R3and R4is HOOC-CH2- and the other independently is (1-4C)alkyl, (1-4C)alkyl-SO2- or HOOC-CH2-; a R5is (3-8C)cycloalkyl, (3-8C)cycloalkyl(1-4C)alkyl, phenyl, benzyl, which is optionally substituted by chlorine or (1-4C)alkoxy. Most preferred are compounds in which a is HOOC-CH2-; B is D-Cha; X is Proline or -[N(cyclopentyl)] -CH2-C(O)-.

If a is R1- SO2-, preferably represents a bond, D-1-Tiq, D-3-Tiq, D-Atc, Aic, D-1-Piq, D-3-Piq or a D-amino acid having a hydrophobic side chain; or a and b taken together represent the residue R3R4N-CHR5-C(O)-, where at least one of R3and R4is R1- SO2- and the other independently is (1-12C) alkyl, or R1- SO2-; X is 2-azetidinone kolovou acid, -[N(3-8C)cycloalkyl]-CH2-C(O) -- or the fragment

< / BR>
or

< / BR>
More preferred are compounds where a is ethyl-SO2- or benzyl-SO2-; B represents a bond, D-Phe, D-Cha, D-Coa, D-Dpa, p-Cl-D-Phe, p-O methyl-D-Phe, p-O-ethyl-D-Phe, D-Nle, m-Cl-D-Phe, 3,4-di-OMe-In-h, D-Chg; or A and B taken together represent the residue R3R4N-CHR5-C(O)-, where at least one of R3and R4represents ethyl-SO2- or benzyl-SO2- and the other independently is (1-12C)alkyl, or R1-SO2-, a R5is (3-8C)cycloalkyl, (3-8C)cycloalkyl(1-4C)alkyl, phenyl, benzyl, diphenylmethyl, where these groups optionally substituted by chlorine or (1-4C)alkoxy. Most preferred are compounds where a is ethyl-SO2; Is D-Cha, and X is Proline or[N cyclopentyl]-CH2-C(O)-.

It is most preferable if the compounds of formula I, r=l.

In the above definitions of the N-protective group radical, And N represents any protective group used in the peptides. Suitable N-protective group can be found in the works of T. W. Green and P. G. M. Wuts: Protective Groups in Organic Synthesis, Second Edition (Wiley, NY, 1991) and in The Peptides, Analysis, Synthesis, Biology, Vol 3, E. Gross and J. Meienhofer, Eds., (AcRaRb-C(O)-, where Raand Rbindependently represent H, a hydrophobic side chain; or Raand Rbtaken together form a 5 - or 6-membered ring which is optionally condensed with one or two aliphatic or aromatic 6-membered rings, and where the aforementioned 5-or 6-membered ring consists of carbon atoms and optionally one heteroatom selected from N, O and S. Preferred D,L-hydroxyazetidine groups are 2-hydroxy-3-cyclohexyl-propionyl and 9-hydroxy-fluoren-9-carboxyl.

The term "(1-12C)alkyl" means a branched or unbranched alkyl group having 1-12 carbon atoms, such as methyl, ethyl, tert-butyl, isopentyl, heptyl, dodecyl, etc. Preferred alkyl groups are (1-6C)-alkyl groups having 1-6 carbon atoms. The most preferred groups mentioned above for R6, R7and R8are (1-3C)alkyl groups having 1-3 carbon atoms, such as methyl, ethyl, isopropyl. (2-12C)Alchemilla group represents a branched or unbranched unsaturated hydrocarbon group having 2-12 carbon atoms. Preferred are (2-6C)alkeneamine group. Examples of such extensive alkylenes group, having 1-6 carbon atoms, such as (CH2)m-, where m=1-6; -CH(CH3)-, -CH(CH3)-(CH2)- etc. Preferred alkionovymi groups in the definition of Y is ethylene and methylene.

(2-12C)Alchemilla group represents a branched or unbranched hydrocarbon group containing a triple bond and having 2 to 12 carbon atoms. Preferred are (2-6C)alkyline groups, such as ethinyl and PROPYNYL.

(6-14C)Aryl group represents an aromatic radical consisting of 6-14 carbon atoms. The aryl group may further contain one or more heteroatoms, such as N, S or O. Examples of aryl groups are phenyl, naphthyl, (ISO)quinoline, indanyl, etc., Most preferred is a phenyl group.

(7-15C)Kalkilya and (8-16C)arlenerina groups are alkyl and alkeneamine group, respectively, substituted by one or more aryl groups, and the total number of carbon atoms in these groups is 7-15 and 8-16, respectively.

The term "(1-6C)alkoxy" means alkoxy group which has 1-6 carbon atoms and the alkyl part of which is such as it was defined above.

The term "halogen" means fluorine, chlorine, bromine or iodine.

The term "ester derivative" means any suitable ester derivative, preferably (1-4C)-alkalemia esters, such as methyl, ethyl or tert-butyl esters.

The term "1 - and 3-Tiq" means 1,2,3,4 - tetrahydroisoquinoline-1 - or 3-carboxylic acid, respectively; the term "1 - and 3-Piq" means the 1 - and 3-carboxypolymethylene respectively; Atc means 2-aminotetralin-2-carboxylic acid; Aic means aminoundecanoic acid; Phe means phenylalanine; Cha means cyclohexylamine; Dpa means diphenylalanine; Coa means cyclooctylamine; Chg means cyclohexylglycine; Nle means norleucine; Asp means aspartic acid.

The term "hydrophobic side chain" means (1-12C)alkyl, optionally substituted by one or more (3-8C) cycloalkenyl groups or (6-14C)aryl groups (which may contain heteroatom, for example nitrogen), for example, such a side chain as cyclohexyl, cyclooctyl, phenyl, pyridinyl, is such substituents as halogen, trifluoromethyl, lower alkyl (e.g. methyl or ethyl), lower alkoxy (e.g. methoxy), phenyloxy, benzyloxy, etc.

In the above definitions, the term "substituted" means substituted by one or more substituents. Examples of amino acids having basic side chain include, but are not limited to, arginine and lysine, preferably arginine. The term "amino acid having a neutral side chain" refers to those amino acids, as methanesulfonic etc.

Examples of cyclic amino acids are 2-azetidinone acid, Proline, pipecolinate acid, 1-amino-1-carboxy-(3-8C)cycloalkyl (preferably 4C, 5C or 6C), 4-piperidinylcarbonyl acid, 4-thiazolidinedione acid, 3,4-dihydropyran, Saproling, 2-octahydrocyclopenta acid, etc., Preferred are 2-azetidinone acid, Proline, pipecolinate acid, 4-thiazolidinedione acid, 3,4-dihydropyran and 2-octahydrocyclopenta acid. The term "Proletarskiy precursor" means a compound in which the nitrogen atom in piperidino group of compounds of formula 1 is secured, for example, hydroxy group, (1-6C)alkoxy group or (1-6C)alkoxycarbonyl Educativa attach suitably protected amino acids or amino acid analogues, with the subsequent removal of the protective groups.

The compounds of formula 1 can be obtained by the method normally used for obtaining such compounds. Modified amino acids having piperidine side chain, is administered in a manner analogous to known methods commonly used for other amino acids.

For this purpose, a suitable derivative or peptide N-protected and protected on the side chain, if present reactive side chains) of amino acids activate and attach to appropriately carboxylterminal the amino acid or peptide derivative either in solution or on solid media. The protection of functional amine groups is usually carried out using a urethane functional groups, such as unstable to acid tert-butoxycarbonyl group (BOC), benzyloxycarbonyl (Z) group and substituted analogs; or unstable to the base 9-fluorenyl-methoxycarbonyl (Fmoc) group. The group Z can also be removed by catalytic hydrogenation. Other suitable aminosidine groups are Nps, Bmv, Rooted, Msc, etc. a Good overview aminosidine groups is given in The Peptides, Analysis, Synthesis, Biology, Vol. 3, E. Gross a, for example unstable to the base esters, such as methyl or ethyl esters; unstable to acid esters, such as tert-butyl ester, or hydrogenations unstable esters, such as benzyl ester. Protection piperidinol side chain can be carried out using the above-mentioned groups. Activation of the carboxyl group suitably protected amino acids or peptides may be carried out using azide, mixed anhydride, active complex ester or carbodiimide, in particular, by adding a catalyst and suppressing racemization compounds such as 1-hydroxybenzotriazole, N-hydroxysuccinimide, 3-hydroxy-4-hydroxy-3,4-dihydro-1,2,3-benzotriazine, N-hydroxy-3 - norbornene-2,3-dicarboxamide. See, for example, Peptides, Analysis, Synthesis, Biology (see above) and Pure and Applied Chem., 59(3), 331-344 (1987).

Compounds of the present invention that may exist in free base form, can be separated from the reaction mixture in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts can also be obtained by treating the free base of formula I with organic or inorganic acid, such as hydrogen chloride, bromide motoroloa, maleic acid, malonic acid, methanesulfonate acid, fumaric acid, succinic acid, tartaric acid, citric acid, benzoic acid and ascorbic acid.

Compounds of the present invention have one or more chiral atoms, and therefore they can be obtained in the form of a pure enantiomer or mixture of enantiomers, or a mixture containing diastereomers. Methods of obtaining pure enantiomers are well known in the art, for example crystallization of salts, which are obtained from optically active acids and racemic mixtures; or chromatography using chiral columns. In the case of diastereomers can be used speakers with direct phase or reversed phase.

Compounds of the present invention can be introduced by oral administration or parenteral, while the preferred daily dose for a human is 0.001 to 100 mg per 1 kg of body weight, and more preferably 0.01 to 10 mg per 1 kg of body weight. Compounds of the present invention, mixed with pharmaceutically acceptable carriers are described, for example, in the famous guide Gennaro et al., Remington''s Pharmaceutical Sciences (18thed., Mack Publishing Company, 1990, see, in particular. Part 8: Pharmaceutical Preparations and Their Manufacture) the ut to be made in the form of capsules or suppositories. Using pharmaceutically acceptable liquid carriers these connections can also be made in the form of a solution, suspension or emulsion, for example, for administration as a drug for injection; or in the form of a spray, for example, for use as aerosols for insertion through the nose.

For the manufacture of dosage forms such as tablets, provides a standard additives, such as fillers, dyes, polymeric binder, etc. basically can be used in any pharmaceutically acceptable additive, provided that it will not have adverse effects on the function of the active compounds. Suitable media that can be entered in the composition are lactose, starch, cellulose derivatives, etc. or mixtures thereof, used in appropriate quantities.

The present invention, also illustrated by the following examples.

Examples

Terms-Ppa[COCO] -, PPP-OMe and Ppa-(2-thiazolyl) mean

< / BR>
where Y represents COOH, OCH3and 2-thiazolyl respectively. Azt = 2-azetidinone acid; Boc = tert-butyloxycarbonyl; Cbz = benzyloxycarbonyl; TsOH = paratoluenesulfonyl acid.

TLC: Rf0,66, silica gel, n-butanol/acetic acid/water, 10/1/3, about/about/about.

(b) N-(t-Butyloxycarbonyl)-D-Cha-OMe

Tert-butylbromide (36 g) was added to a stirred solution of H-D-Cha-OMe. HCl (40,9 g) in 400 ml of acetonitrile. the pH of the mixture was brought to 8.5 by adding N, N-diisopropylethylamine. The mixture was stirred for 16 hours at room temperature and evaporated in vacuum. The residue was dissolved in dichloromethane and the solution washed with water, dried with sodium sulfate and evaporated in vacuum. After chromatography on silica gel using as eluent heptane/ethyl acetate, 9/1, V/V, was obtained 64 g of N-(t-butyloxycarbonyl)-D-Cha-OMe.

TLC: Rf0.25, silica gel, ethyl acetate/heptane, 1/1, V/V.

(C) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-OMe

the pH of a solution of N-(t-Butyloxycarbonyl)-D-Cha-OMe (64 g) and di-t-BUTYLCARBAMATE (40,3 g) was brought to 8.5 by adding N,N-diisopropylethylamine was added dichloromethane and water. The organic layer was separated, washed with cold 1 n hydrochloric acid, water, 5% sodium bicarbonate and again with water, and then dried with sodium sulfate, and the filtrate was evaporated to obtain N-(t-butyloxycarbonyl)-N-BOC-D-Cha-OMe in the form of amorphous solids with access to 59.6 g

TLC: Rfof 0.50, silica gel, ethyl acetate/heptane, 1/1, V/V.

(d) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-OH

A solution of N- (t-butyloxycarbonyl) -N-Boc-D-Cha-OMe (59,6 g) in 900 ml of dioxane/water (9/1, V/V) was treated with a sufficient amount of 6 n sodium hydroxide to maintain pH = 12 for 6 hours at room temperature. After acidification, the mixture was poured into water and was extracted with dichloromethane. The organic layer was washed with water and dried with sodium sulfate. The filtrate was evaporated and obtained 54 g of N-(t - butyloxycarbonyl)-N-Boc-D-Cha-OH.

TLC: Rf0,60; silica gel, dichloromethane/methanol, 9/1, V/V.

(e) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-OBzl

To a cold (0oC) a solution of N-(t-butyloxycarbonyl)-N-Boc-D-Cha-OH (13.5 g) in N,N-dimethylformamide (150 ml) was sequentially added 1-hydroxybenzotriazole (HOBT) (7.09 in), dicyclohexylcarbodiimide (DCCl) (to 7.61 g), H-Pro-Obzl. HCl (9,31 g) and triethylamine (6 ml). The mixture was stirred whether to -20oC, and dicyclohexylamine was removed by filtration. The filtrate was evaporated to dryness. The residue was dissolved in ethyl acetate and sequentially washed with 5% sodium bicarbonate, water, 3% citric acid and saturated saline, then was dried with sodium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel, using as eluent heptane/ethyl acetate, 3/1, /about/about. The fractions containing N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-zl, were combined and evaporated. The yield was 15 g

TLC: Rf0,70, silica gel, heptane/ethyl acetate 1/1 V/V.

(f) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-OH

To a solution of N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-OBzl (15 g) in methanol (150 ml) was added 10% palladium on coal (750 mg). The mixture was first made at atmospheric pressure and at room temperature for 1 hour. The palladium catalyst was removed by filtration and the solvent was removed by evaporation under reduced pressure, resulting in an 11.2 g of N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro-OH.

TLC: Rf0,65, silica gel, ethyl acetate/pyridine/acetic acid/water, 213/20/6/11, about/about/about/about.

(g) of Diethyl 2-acetamido-2-[(pyridin-4-yl)methyl]-malonate

Portion (46 g) m is ethylacetamide (217 g) and 4-picolylamine hydrochloride (164 g). The reaction mixture was heated to 100oC for 4 hours, cooled, filtered and concentrated in vacuum. The residue was dissolved in dichloromethane, filtered through diesel and silica gel and concentrated in vacuum. The residue was treated with 1 l of diethyl ether, filtered and washed with diethyl ether. Output: 142,

TLC: Rf0.5 ethyl acetate/pyridine/acetic acid/water, 63/5/1,5/2,75, about/about/about/about, on silica gel.

(h) Ethyl 2-acetamido-2-[(pyridin-4-yl)methyl]-malonate

A solution of 142 g of diethyl 2-acetamido-2-[(pyridin-4-yl)methyl]-malonate, 46 g of potassium hydroxide in 2 liters of ethanol and 800 ml of water was stirred for 16 hours. The mixture was acidified and concentrated in vacuum. The co-evaporation with methanol and toluene were received 187 g of ethyl 2-acetamido-2-[(pyridin - 4-yl)methyl]-malonate (containing potassium chloride).

TLC: Rf0,95 on silica gel; ethyl acetate/pyridine/acetic acid/water, 63/5/1,5/2,75, about/about/about/about.

(i) Ethyl 2-N-acetylamino-3-(pyridin-4-yl)propanoate

A solution of 394 g of ethyl 2-acetamido-2- [(pyridin-4-yl)methyl]-malonate in 1.5 liters of N, N-dimethylformamide was heated under reflux for 1.5 hours and concentrated in vacuum. The residue was dissolved in water and the pH brought to 11.5. CME is TLC: Rf0,35, ethyl acetate/pyridine/acetic acid/water, 63/5/1,5/2,75, about/about/about/about, on silica gel.

(j) Ac-Ppa-OEt

A solution of 39.5 g of ethyl 2-N-acetylamino-3- (pyridin-4-yl)propanoate in 1 l of ethanol was restored in an atmosphere of hydrogen using 4 g of 10% palladium on charcoal as catalyst. After 18 hours the catalyst was removed by filtration and the filtrate was concentrated to dryness to obtain 40 g of the desired compound.

(k) H-Ppa-OH

A solution of 81.5 g of Ac-Ppa-OEt in 6 n hydrochloric acid was heated under reflux for 4 hours. The reaction mixture was concentrated in vacuum and received 90 g H-Ppa-OH.HCl.

TLC: Rf0,97, 1-butanol/pyridine/acetic acid/water, 8/3/1/4, about/about/about/about, on silica gel.

(l) Cbz-Ppa(Boc)-HE

Brought the pH of a solution of 90 g of H-Ppa-OH, 51,23 g of the pentahydrate of copper sulfate (II) and 166 g of di-t-buildinternet in water (1730 ml) and dioxane (1250 ml) to the value of 9 by adding 2 n sodium hydroxide. The mixture was stirred for 16 hours at room temperature. The precipitate was collected and thoroughly washed with water. The filter cake was dissolved in dioxane, and the pH was brought up to 12.5 by addition of 4 n sodium hydroxide was added to 200 g of benzyloxycarbonylglycine. A mixture of paramesh and the pH was brought to 2.5. The residue was diluted with ethyl acetate. The organic layer was washed with water, dried with sodium sulfate and was evaporated to dryness to obtain 160 g of target compound.

TLC: Rf0,77; ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about, on silica gel.

(m) Cbz-Ppa(Boc)-OMe

To a solution of 160 g of Cbz-Ppa(Boc)-HE's in dichloromethane/methanol (9/1, V/V) (3 l) was added 120 g of tetrafluoroborate 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea (TBTU), after which the pH was brought to 8.5 by the addition of triethylamine. The resulting mixture was stirred for 1 hour at room temperature. The organic layer was washed with 2 n hydrochloric acid, water, 5% sodium bicarbonate solution and again with water, then dried with sodium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel using as eluent dichloromethane/ethyl acetate, 9:1, vol/vol. The fractions containing methyl ester were combined and the solvent was removed in vacuum. The residue was led from diisopropyl ether/diethyl ether, and received the first collection of 75 g of the desired product. The mother liquor was chromatographically in toluene/ethyl acetate, 8/2, V/V and got 34 g of Cbz-Ppa(Boc)-OMe.

TLC: Rf0,77, dichloromethane/methanol, 95/5, V/V, at the Rhyl

At -78oC 908 ml of pre-cooled hydride diisobutylaluminum (1.0 M solution in hexane) was added to a stirred solution of 109 g of Cbz-Ppa(Boc)-OMe in dichloromethane (3 l) so that the temperature was maintained below -70oC. the Solution was stirred for 1 hour at -70oC, and then added 1 l of a saturated solution of citric acid. The mixture was poured into two liters of a solution of citric acid and was extracted with dichloromethane. The combined extracts were washed with water, 5% sodium bicarbonate solution and again with water, and then dried with sodium sulfate and filtered. The filtrate was cooled to 0oC and sequentially added 15 g of the chloride of benzyltriethylammonium, 58 ml of acetic anhydride and a solution of 118 g of sodium cyanide in water (1.5 l). The resulting mixture was intensively stirred for 30 minutes. The organic layer was separated, washed twice with water, dried with sodium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel, elwira hexane/ethyl acetate, 6/4, V/V, resulting in a received target compound in the form of solids: 98,

TLC: Rf0,65, dichloromethane/methanol, 95/5, V/V on silica gel.

(a) Cbz-Ppa[CHOHCO]-OMe

Stir a suspension of 98 g of 2 - acetoxy-3-(benzilic is b) was treated with gaseous hydrogen chloride (260 g) at -20oC. the Mixture was stirred for 24 hours at 0-5oC. Then the mixture was cooled to -20oC and added 408 ml of water. The reaction mixture was stirred for 4 hours at -20oC and the organic layer was separated. the pH of the aqueous layer was brought to 8 by addition of 25% aqueous ammonia, and then this layer was extracted with 1-butanol. The combined extracts were washed with saturated salt solution, dried with sodium sulfate and was evaporated to dryness to obtain 57 g of Cbz-Ppa[CHOHCO]-OMe.

TLC: Rf0.5 on silica, ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about.

(b) Cbz-Ppa(Boc)[CHOHCO]-OMe

the pH of the solution of Cbz-Ppa[CHOHCO]-OMe (57,9 g) and 40 g of di-tributylphosphate in 500 ml of N,N-dimethylformamide was brought to 8.5 by the addition of triethylamine. The mixture was stirred for 45 minutes at room temperature. The solvent was removed in vacuum. The residue was dissolved in ethyl acetate, and then washed with 0.1 n hydrochloric acid, water, 5% sodium bicarbonate and again with water. The organic layer was dried with sodium sulfate and evaporated to dryness. The residue was chromatographically on silica gel, elwira heptane/ethyl acetate, 6/4, V/V, and received 31 g of oil.

(q) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[CHOHCO]-OMe

A solution of 120 mg of Cbz-Ppa(Bo is atora in N,N-dimethylformamide (4 ml) was first made at atmospheric pressure. After the reaction was completed, the catalyst was removed, a pH of the solution was brought to 8.5 by the addition of triethylamine. The resulting solution was added to a stirred solution of 137 mg of N-(t-Butyloxycarbonyl)-N-BOC-D-Cha-Pro-HE, 40 mg of 1-hydroxybenzotriazole and 61 mg of 1,3-dicyclohexylcarbodiimide in 10 ml of N,N-dimethylformamide. The reaction mixture was stirred for 16 hours at room temperature. The solvent was removed in vacuo and to the residue was added dichloromethane. The precipitate was filtered and the filtrate washed with 1 n hydrochloric acid, water, 5% sodium bicarbonate solution and again with water. The organic layer was dried with sodium sulfate and evaporated in vacuum to obtain 220 mg of the desired compound.

TLC: Rf0.6 silica gel, ethyl acetate/pyridine/acetic acid/water, 63/5/1,5/2,75, about/about/about/about.

(r) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-PPA(Boc)[CHOHCO]-

the pH of the solution of 220 mg of N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-Ppa(Boc) [CHOHCO]-OMe in 15 ml of dioxane/water (9/1, V/V) was brought to 12 by adding 1 n sodium hydroxide. The reaction mixture was stirred for one and a half hours. the pH was brought to 2.5 by adding 1 n hydrochloric acid and the mixture was poured into water and then was extracted with ethylacetate.

TLC: Rf0,21 on silica gel, ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about.

(s) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[COCO]-OH

To a solution of 210 mg (r) N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-Ppa(Boc)[CHOHCO] -OH in 20 ml of dichloromethane was added 177 mg 1,1,1-triacetoxy-1,1-dihydro-1,2-benzodioxol-3(1H)-she (reagent dess-Martin). The mixture was stirred for 4 hours at room temperature, was poured into a solution of sodium thiosulfate (5% in water) and was extracted with dichloromethane. The organic layer was washed with water, dried with sodium sulfate and concentrated in vacuum. Yield: 340 mg

TLC: Rfof 0.4 on silica gel, ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about.

(t) HOOC-CH2-D-Cha-Pro-Ppa[COCO]-

A solution of 340 mg of N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro-PPa(Boc)[COCO]-HE in 20 ml of 90% triperoxonane acid was stirred for 4 hours at room temperature. The solvent was removed in vacuo and the residue was purified on preparative HPLC-column LC-18-BD Supelcosil using gradient elution: 20%; B: 80%; 0% ---> A: 20%; In: 55%; C: 25% for 45 minutes at a flow rate of 20 ml/min (A: 0.5 M phosphate buffer with pH 2,1; B: water: acetonitrile:water, 3:2, V/V), resulting received ptx2">

Example 2

HOOC-CH2-D-Cha-N-cyclopentyl-Gly-PPA[COCO]-

(a) N-Cyclopentyl-Gly-OMe.HCl

H-Gly-OMe. HCl (46.4 g) was dissolved in 400 ml of methanol, and then was added Cyclopentanone (34 g) and cyanoborohydride sodium (14 g) and the reaction mixture was left for 16 hours at room temperature to complete the reaction. The reaction mixture was extinguished 6 M hydrochloric acid until then, until the pH does not become equal to 2, and then was stirred for 30 minutes at room temperature. The solvent was removed by evaporation under reduced pressure, and the residue was dissolved in water and washed with diethyl ether. The PH is brought up to > 10 by addition of 6 M sodium hydroxide solution, whereupon the product was extracted with dichloromethane, washed with saturated saline solution, dried with sodium sulfate, filtered and concentrated under reduced pressure. The compound was crystallized from ethyl acetate as HCl-salt. Output: 43,5 g

TLC: Rf0,71; silica gel; ethyl acetate/pyridine/acetic acid/water, 88/31/18/7, about/about/about/about.

(b) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-N-cyclopentyl-Gly-O

N-(t-Butyloxycarbonyl)-N-BOC-D-Cha-N-cyclopentyl-Gly-OH was obtained by a method similar to that described in Example 1 for dipeptide part,Bt-linking between N-(t-butyloxycarbonyl)-N-BOC-D-Cha-N-cyclopentyl-Gly-HE and H-Ppa(Boc)[CHOHCO] - OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, resulting in the obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 29,72 min; 20%A/80% - 20%A/20%/ 60%To 40 minutes.

Example 3

HOOC-CH2-D-Phe-Pro-Ppa[COCO]-OH

N-(t-Butyloxycarbonyl)-N-Boc-D-Phe-Pro-OH was obtained as described in Example 1, on the basis of H-D-Phe-OH.HCl.

DCCl/HOBt-coupling between N-(t-butyloxycarbonyl)-N-Boc-D-Phe-Pro-OH and H-Ppa(Boc)[CHOHCO] -OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, resulting in the obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 17,77 and 18,19 min; 20%A/80% - 20%A/20%/60%To 40 minutes.

Example 4

HOOC-CH2-m-Cl-D,L-Phe-Pro-PPA[COCO]-

H-m-Cl-D, L-Phe-OH. HCl was obtained from 3-chlorobenzylamino the method described in Example 53. Then the Assembly is fully protected Tripeptide carried out by methods similar to

described in Example 1. The target compound was obtained as a mixture of diastereoisomers.

Rt(LC): 22,24/23,07 min; 20%A/80% - 20%A/20%/60%To 40 minutes.

Example 5

(HOOC-CH2-p-Cl-D-Phe-Pro-Ppa[COCO]-OH

(a) N-(t-Butyloxycarbonyl)-N-Boc-p-Cl-D-Phe-OH. Output: 16,7,

TLC: Rf0,27; silica gel, ethyl acetate/methanol, 9/1, V/V.

(b) N-(t-Butyloxycarbonyl)-N-Boc-p-Cl-D-Phe-OSu (Su=succinimide)

A solution of N-(t-Butyloxycarbonyl) -N-Boc-p-Cl-D-Phe-OH (14,67 g) in 250 ml of acetonitrile was treated with N-hydroxysuccinimide (4.11 g) and the hydrochloride of 1-(3 - dimethylaminopropyl)-3-ethylcarbodiimide (6,86 g) overnight at room temperature. The reaction mixture is evaporated to dryness and the residue was dissolved in ethyl acetate. The organic phase is washed with water, dried with sodium sulfate and concentrated to obtain 19,11 g of the active complex ester which was used directly in the subsequent stage.

(C) N-(t-Butyloxycarbonyl)-N-Boc-p-Cl-D-Phe-Pro-OH

H-Pro-OH.HCl (10,79 g) was dissolved in 100 ml of N,N-dimethylformamide and 100 ml of water. the pH of the reaction mixture was brought to 8 by adding 1 n sodium hydroxide solution, and then was added dropwise N-(t-butyloxycarbonyl)-N-BOC-p-Cl-D-Phe-OSu (19,11 g), dissolved in 120 ml of N,N-dimethylformamide. The reaction mixture was stirred over night at room temperature at pH ~ 8. This reaction mixture was cooled and brought to pH=2 by adding 1N hydrochloric acid. The aqueous layer was extracted with dichloromethane. Organicheskoyj gradient elution of ethyl acetate/methanol, 9/1 - 1/1, V/V, was received? 7.04 baby mortality g of the desired dipeptide.

TLC: Rf0,24; silica gel, ethyl acetate/methanol 8/2, about/about.

(d) HOOC-CH2-p-Cl-D-Phe-Pro-Ppa[COCO]-

DCCl/HOBt - coupling between N-(t-butyloxycarbonyl)-N-BOC-p-Cl-D-Phe-N-cyclopentyl-Gly-HE and H-Ppa(Boc)[SENSE]-OMe.HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1. The target compound was obtained as a mixture of diastereoisomers.

Rt(LC): 23,67 min 20%A/80%B To 20%A/20%B/ 60%C in 40 minutes.

Example 6

NOOS-CH2-D-p-MeO-Phe-Pro-Ppa[COCO]-

By the way, is similar to that described in Example 1, from N-p-MeO-D-Phe-OH was obtained target Tripeptide as diastereomeric mixture.

Rt(LC): MT 19: 18 min 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 7

HOOC-CH2-D,L-Coa-Pro-Ppa[COCO]-OH

(a) Cyclooctylmethyl

Cyclooctylmethyl (8,16 g) was dissolved in 47% HBr-solution (70 ml) and heated under reflux for 1 hour at 130oC. the Reaction mixture was poured into ice water (500 ml) was added a saturated solution of sodium bicarbonate (500 ml). The aqueous solution was extracted with dichloromethane. The combined organic phases are washed with water and saturated salebrowse on silica gel, using as eluent toluene. The fractions containing cyclooctylamine, were combined and evaporated. Output: 9,85,

TLC: Rf0,95; silica gel, toluene.

(b) (R,S)-Ethyl-2-acetylamino-2-cyano-3-cyclooctylamine

Tert-butyl potassium (6.85 g) and ethylacetoacetate (8.1 g) was dissolved in dimethyl sulfoxide (100 ml) at room temperature. Cyclooctylmethyl was dissolved in dimethyl sulfoxide (25 ml) and was added dropwise to the reaction mixture. The mixture was stirred at room temperature for 44 hours. After pouring 500 ml of water the precipitate was filtered and dried, resulting in the received (R, S)-ethyl-2-acetylamino-2-cyano-3-cyclooctylamine (2,95 g).

TLC: Rfof 0.50, silica gel, heptane/ethyl acetate 3/7, about/about.

(C) H-D,L-Coa-OH.HCl

(R, S)-Ethyl-2-acetylamino-2-cyano-3 - cyclooctylamine (2,95 g) suspended in 100 ml of 20% hydrochloric acid and heated under reflux for 22 hours. The reaction mixture was cooled to 5oC and the resulting precipitate was filtered, washed with diethyl ether and dried. Output: 2,69 g H-D,L-Coa-OH.HCl

TLC: Rf0,27; silica gel; ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about.

HOOC-CH2-D, L-Coa is diastereomers.

Rt(LC): 15,19 min; 20%A/60%/20% And - 20%A/80%C for 30 minutes.

Rt(LC): 25,71 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 8

HOOC-CH2-D-Nle-Pro-Ppa[COCO]-OH

Target Tripeptide as a mixture of diastereoisomers was obtained by the method described in Example 1, on the basis of H-D-Nle-OH.

Rt(LC): 19,50 and 20,24 min; 20%A/80% - 20%/50%/30%C for 40 minutes.

Example 9

HOOC-CH2-D-Dpa-Pro-Ppa[COCO]-OH

By the way, is similar to that described in Example 1, on the basis of H-D-Dpa-OH was obtained target compound, Tripeptide, in the form of a mixture of diastereoisomers.

Rt(LC): 29,18 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 10

D-Hpl-Pro-Ppa-(2-thiazolyl) (Hp1=2-hydroxy-3-phenylmalonate acid)

(a) THP-D-Hpl-Pro-OH

Synthesis of THP-D-Hpl-Pro-OH, on the basis of H-D-Cha-OH, described in Example 20.

(b) D-Hpl-Pro-Ppa-(2-thiazolyl)

The reaction of binding of THP-D-Hpl-Pro-OH with H-Ppa(Cbz)-(2-thiazolyl) TFA, followed by the removal of protection and purification was performed as described in Example 45, resulting in a received D-Hpl-Pro-Ppa-(2-thiazolyl) as a mixture of diastereoisomers.

Rt(LC): 23,42 and 24,36 min; 20%A/60%b and 20%C 100%C for 40 minutes.

Example 11

HOOC-CH2-u-Me-D-Phe-Pro-PPA[COCO]-

(a) Fmoc-n-Me-D-Phe-OMe

Fmoc-n-Me-D-Phe-OH (5) - Rev.,3-tetramethylurea (4.4 g) the solution was brought to pH 8 by the addition of triethylamine. The reaction mixture was stirred for 16 hours at room temperature. The mixture is then washed with cold 1 n hydrochloric acid, water, 5% sodium bicarbonate and again with water, and then dried with sodium sulfate. The filtrate was evaporated and obtained 5.6 g Fmoc-n-Me-D-Phe-OMe in the form of butter.

TLC: Rfof 0.55, silica gel, ethyl acetate/heptane, 6/4, about/about.

(b) N-n-Me-D-Phe-OMe

Fmoc-n-Me-D-Phe-OMe (5.6 g) was dissolved in 35 ml of N, N-dimethylformamide/piperidine (4/1) and was stirred for 30 minutes at room temperature. The mixture was evaporated and the residue was dissolved in ethyl acetate and washed with 0.1 M hydrochloric acid, the pH of the combined aqueous layers were brought to 9 by adding 1 M sodium hydroxide solution. After extraction with dichloromethane the resulting oil was chromatographically on silica gel, elwira ethyl acetate/methanol, 95/5, about/about. The fractions containing N-n-Me-D-Phe-OMe, were combined and evaporated. Output: 2,33,

TLC: Rf0,10, silica gel, heptane/ethyl acetate 3/2, about/about.

Then spent the Assembly fully protected Tripeptide ways, similar to that described in Example 1. The target compound was identified as a mixture of diastereoisomers.

Rt(LC): 22,55 min; 20%A/80% - 20%A/20%Gly means a structural fragment of the formula:

< / BR>
(a) 3-(piperidine-4-yl)propanoic acid

A mixture of 12.5 g of 3-(4-pyridyl)acrylic acid and 1.25 g of 10% Pd/C as catalyst in 1 l of ethanol and 83 ml of 1 n hydrochloric acid was shaken in a hydrogen atmosphere for 64 hours. The catalyst was removed by filtration and the filtrate was evaporated to dryness to obtain 13 g of the desired compound.

TLC: Rf0,7; silica gel; butanol/pyridine/acetic acid/water, 4/1/1/2, about/about/about/about.

(b) 3-[1-(t-butyloxycarbonyl)piperidine-4-yl]propanoic acid

A suspension of 3-(piperidine-4-yl)propanoic acid in 500 ml of N,N-dimethylformamide was treated 30,9 g of di-t-BUTYLCARBAMATE, the pH of the reaction mixture is maintained 8,5 using triethylamine and then stirred for 4 hours at room temperature. The mixture was poured into 65 ml of chilled 2 n sodium hydroxide solution and washed with ethyl acetate. The aqueous phase was acidified to pH=2 by adding 2 n hydrochloric acid and was extracted with ethyl acetate. The combined organic layers were washed with saturated salt solution, dried with sodium sulfate and evaporated in vacuum to obtain 37 g of the desired compound.

TLC: Rf0,7, silica gel, dichloromethane/methanol, 9/1, V/V.

(C) (4S)-3-[3-(t-butyloxycarbonyl)piperidine-4-yl]propanoic acid in 325 ml of dry tetrahydrofuran was carried out by treatment of 13.7 ml pivaloyloxy and 15.6 ml of triethylamine at -72oC. the Mixture was stirred for 1 hour at 0oC. To a solution of 18.5 g 3-4 - benzyl-2-oxazolidinone in 325 ml of dry tetrahydrofuran at -72oC in an atmosphere of nitrogen was added dropwise 75 ml of 1.4 M solution of n-BuLi in hexane. The mixture was stirred for 20 minutes at -70oC. After addition of the suspension of the above mixed anhydride the reaction mixture was stirred another 20 minutes at -70oC. the Reaction is extinguished by pouring into water and the mixture was extracted with ether. The combined organic layers were washed with cold 1 n hydrochloric acid, 5% sodium bicarbonate solution and water. The solution was dried with sodium sulfate and evaporated in vacuum to obtain and 47.5 g of oil. After flash chromatography on silica gel using as eluent heptane/ethyl acetate (1:1, V/V) was received of 28.2 g of target compound.

TLC: Rf0,7, heptane/ethyl acetate, 1:1, V/V on silica.

(d) (3(2S), 4S)-3-[2-Azido-3-[1-(t-butyloxycarbonyl) piperidine-4-yl]-1-oxo-propyl]-4-(phenylmethyl)-2-oxazolidine

To 110 ml of dry tetrahydrofuran, stirred at -78oC in nitrogen atmosphere, was added 136 ml of 0.5 M hexamethyldisilazide potassium in toluene. To the resulting solution through a cannula was added to the pre-ohlord is Liliana in 350 ml of dry tetrahydrofuran and stirring was continued for 30 minutes at -75oC in nitrogen atmosphere. To the above solution enolate potassium, stir at -75oC, was added a solution of 24 g trailside (lit. JCS Perkin Trans (1991), 1629) in 215 ml of dry tetrahydrofuran. After 5 minutes, the reaction was suppressed by adding 17 ml of acetic acid. The mixture was stirred for 20 minutes at - 75oC and for 16 hours at room temperature.

After flash chromatography on silica gel using as eluent heptane/ethyl acetate (7/3, V/V) and after treatment with heptane was obtained 19 g of target compound in the form of a solid substance.

TLC: Rf0,6, silica, dichloromethane/ethyl acetate, 9/1, V/V.

(e) (2S)-2-Azido-3-[1-(t-butyloxycarbonyl)piperidine-4-yl] -propanoic acid

To a solution of 19 g (3(2S), 4S)-3-[2-azido-3-[1-(t-butyloxycarbonyl)piperidine-4-yl] -1-oxo-propyl]-4-(phenylmethyl)-2-oxazolidinone in 600 ml of tetrahydrofuran (THF, 18 ml of 30% hydrogen peroxide and 170 ml of water was added 3.7 g of solid lithium hydroxide at 0oC in nitrogen atmosphere. The mixture was stirred for 30 minutes at 0oC in nitrogen atmosphere. Then was added a solution of 25.3 g of sodium sulfite in 133 ml of water and 530 ml of 5% sodium bicarbonate solution. The temperature was raised to room temperature and stirring prodajem. The aqueous phase was acidified using 6 n hydrochloric acid to pH 2 and extracted with ethyl acetate. The extracts were washed with saturated salt solution, dried with sodium sulfate and evaporated in vacuum. After crystallization of the residue from ether/heptane was obtained 11.9 g of crystalline solids.

TLC: Rf0,8, silica, dichloromethane/methanol = 95/5, V/V.

(f) (2S)-N-Methyl-N-methoxy-2-azido-3-[1-(t - butyloxycarbonyl)piperidine-4-yl]-propanamide

To a solution of 5.7 g of (2S)-2-azido-3-[1-(t-butyloxycarbonyl)piperidine-4-yl] -propanoic acid in 150 ml of dichloromethane was added 2.1 g of N,O-dimethylhydroxylamine. HCl and 6.4 g of TBTU. the pH was maintained at 8,5 using triethylamine. The mixture was stirred for 30 minutes at room temperature, and then washed with 1 n hydrochloric acid, water, 5% sodium bicarbonate and again with water. The organic layer was dried with sodium sulfate, filtered and evaporated in vacuum to obtain 7.4 g of the target compound in the form of butter.

TLC: Rf0,6, silica, dichloromethane/methanol, 95/5, about/about.

(g) Methyl (23)-2-azido-3-[1-(t-butyloxycarbonyl)piperidine-4-yl]-propanoic acid

the pH of a mixture of (2S)-2-azido-3-[1-(t-butyloxycarbonyl)piperidine-4-yl]- propane lamina. The mixture was stirred for 30 minutes at room temperature, and then washed with 1N hydrochloric acid, water, 5% sodium bicarbonate solution and again with water. The organic layer was dried with sodium sulfate, filtered, evaporated in vacuum and got to 7.4 g of target compound in the form of butter.

TLC: Rf0,85, silica, dichloromethane/methanol = 95/5, V/V.

(h) H-S -- Ppa(Boc)-OMe

A solution of methyl(2S)-2-azido-3-[1-(t-butyloxycarbonyl)piperidine-4-yl] -propanoic acid (7.4 g), 0.5 g of Pd/C (10%) and 21.9 ml of 1 n hydrochloric acid in tetrahydrofuran (300 ml) was first made at atmospheric pressure. The catalyst was filtered, and the solvent was removed in vacuum and after treatment with ether there was obtained 6.8 g of crystalline solids.

TLC: Rf0,85; ethyl acetate/pyridine /acetic acid/water, 252/20/6/11, about/about/about/about.

(i) Cbz-S-Ppa(Boc)-OMe

the pH of the solution of 6.5 g H-S -- Ppa(Boc)-OMe and 5.6 g of benzyloxycarbonylglycine in 70 ml of N, N-dimethylformamide was brought to 8.5 using triethylamine. The mixture was stirred for 6 hours at room temperature, poured into water and was extracted with ethyl acetate. The combined organic layers washed with 1 n hydrochloric acid, water, 5% to the left connection in the form of oil.

TLC: Rf0,85, silica, heptane/ethyl acetate, 1:1, about/about.

(j) Cbz-S-Ppa(Boc)-H

To a solution of 7.5 g of Cbz-S-Ppa(Boc)-OMe in 225 ml of dry dichloromethane at -75oC in an atmosphere of nitrogen was added dropwise to 68.1 ml of 1 M solution of hydride diisobutylaluminum in hexane while maintaining the temperature below -70oC. the Mixture was stirred for one hour at -75oC in nitrogen atmosphere. This mixture was poured into a solution of citric acid and was extracted with ethyl acetate. The combined extracts were washed with water, 5% sodium bicarbonate solution and water, and then concentrated in vacuum to obtain 7.5 g of the target compound.

TLC: Rf0,45, dichloromethane/methanol (95/5, V/V) on silica.

(k) (3S)-2-Acetoxy-3-(benzyloxycarbonylamino)-4-[1-t-butyloxycarbonyl(piperidine-4-yl)]-butanetriol

Cbz-S-Ppa(Boc)-H (7.5 g) (Example 12 (j)) are transformed into (3S)-2-acetoxy-3-(benzyloxycarbonylamino)-4-[1-t-butyloxycarbonyl(piperidine-4-yl)] -butanetriol manner similar to that described in Example 1(n).

TCX: Rf0,75, dichloromethane/methanol (95/5) (V/V) on silica.

(1) Cbz-S-Ppa(Boc)[CHOHCO]-OMe

(3S)-2-Acetoxy-3-(benzyloxycarbonylamino)-4-[1-t-butyloxycarbonyl (piperidine-4-yl)] -butanetriol become rmean/methanol, 95/5, about/about.

(m) H-S -- Ppa(Boc)[CHOHCO]-OMe.HCl

H-S -- Ppa(Boc)[CHOHCO] -OMe.HCl was obtained by way of hydrogenation as described in Example 1 (q)

(n) N-BOC-L-Amino-caprolactam

To a stirred solution of L-amino-caprolactam (10 g) in dioxane/water (2: 1, V/V) (30 ml) was added 1 n sodium hydroxide solution (7.8 ml) and then di-tert-butyl dicarbonate (18,8 g). The mixture was stirred for 16 hours at room temperature and concentrated in vacuum. The residue was dissolved in ethyl acetate, washed with water and saturated saline, and then dried with sodium sulfate, filtered and evaporated in vacuum. The crude product is triturated with hexane, filtered and dried in vacuum to obtain N-BOC-L-amino-caprolactam (16 g).

TCX: Rf0,85, on silica, ethyl acetate/heptane, 1/1, V/V.

(a) Boc-norLeu(cyclo)-Gly-OMe

N-Boc-L-Amino-caprolactam (10 g) was dissolved in dichloromethane (9100 ml). Then at -20oC was slowly added a 1M solution of bis(trimethylsilyl)amide in tetrahydrofuran/cyclohexane (1: 1, V/V) (1 equiv.) and the mixture was stirred for 30 minutes. After adding methylpropanoate (4 ml) and the mixture was stirred for 2 hours at room temperature. To expedite completion of the reaction was added additional quantities of the 0.1 n solution of hydrochloric acid, water, 5% aqueous sodium bicarbonate solution and saturated saline, and then dried with sodium sulfate, filtered and evaporated in vacuum. The residue was purified by chromatography on silica (eluent: heptane/ethyl acetate=6:4, V/V) and were obtained 12 g of Boc-norLeu(cyclo)-Gly-OMe

TLC: Rf0,55, ethyl acetate/heptane, 6/4, V/V on silica.

(b) Benzyl SO2-norLeu(cyclo)-Gly-OMe

Boc-norLeu(cyclo)-Gly-OMe (3 g) was dissolved in TFA/dichloromethane (1:1, V/V) (30 ml) and was stirred for one hour at room temperature. The reaction mixture was evaporated in vacuum. The crude amine was dissolved in dichloromethane (25 ml) slowly at 0oC solution was added benzylchloride (2.25 g) in dichloromethane (10 ml). Then was added triethylamine to maintain pH=8 throughout the reaction. The mixture was stirred for 1 hour at room temperature, and then concentrated in vacuum. The residue was dissolved in ethyl acetate, washed with 5% sodium bicarbonate solution, water and saturated saline, and then dried with sodium sulfate, filtered and evaporated in vacuum. The residue was purified by chromatography on silica (eluent: dichloromethane/ethyl acetate = 95:5, V/V) and was obtained benzyl SO2-norLeu(CEC is Benzyl SO2-norLeu(cyclo)-Gly-HE

A solution of benzyl SO2-norLeu(cyclo)-Gly-OMe (3,9 g) in 100 ml of dioxane/water (9:1) was treated with a sufficient amount of 1 n sodium hydroxide to maintain pH= 13 for 2 hours at room temperature. After acidification, the mixture was poured into water and was extracted with dichloromethane. The organic layer was washed with water and dried with sodium sulfate. The filtrate was evaporated and obtained 3.6 g of the target compound.

TLC: Rf0,60; ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about, silicon dioxide.

(r) Benzyl SO2-norLeu(cyclo)-Gly-S-Ppa[COCO]-

DCCl/HOBt-coupling between benzyl SO2-norLeu(cyclo)-Gly-HE H-S -- Ppa(Boc)[CHOHCO]-OMe.HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1.

Rt(LC): 27,41 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 13

1-Piq-Pro-Ppa[COCO]-

N-(Cbz)-1-Piq-OH

N-(Cbz)-1-Piq-OH was synthesized by the method described in EP 0643073 in Example 1.

TLC: Rf0,85; ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about, silicon dioxide.

(a) N-(Cbz)-1-Piq-Pro-OtBu

To a cold solution (0oC) N-(Cbz)-1-Piq-OH (500 mg) in N,N-dimethylformamide (95 ml) was sequentially added DCCl (342C, and then stood overnight at room temperature. The reaction mixture was cooled to -20oC and DCU (1,3-dicyclohexylphosphino) was removed by filtration. The filtrate was concentrated in vacuo, and the residue was dissolved in ethyl acetate. This solution was sequentially washed with 5% aqueous solution of sodium bicarbonate, 3% aqueous citric acid solution, water and saturated salt solution. And then was dried with sodium sulfate and concentrated in vacuum. The residue was purified by chromatography on silica (eluent: heptane/ethyl acetate, 4/1, V/V) and was obtained N-(Cbz)-1 - Piq-Pro-OtBu (634 mg).

TLC: Rf0,90; ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about, silicon dioxide.

(b) N-(Cbz)-1-Piq-Pro-OH

N-(Cbz)-l-Piq-Pro-OtBu (600 mg) was stirred in a mixture of dichloromethane (1 ml), triperoxonane acid (3 ml) and anisole (0.15 ml) for one hour at room temperature. The reaction mixture was concentrated in vacuo at low temperature and the residue was dissolved in water at pH = 9.5. The aqueous phase is washed with diethyl ether and then the aqueous layer was acidified to a pH of 2.5 by adding 2 M hydrochloric acid. The aqueous layer was extracted with ethyl acetate, and the organic phase was washed us is).

TLC: Rf0,54; ethyl acetate/pyridine/acetic acid/water, 60/3/1/2, about/about/about/about, silicon dioxide.

(c) 1-Piq-Pro-Ppa[COCO]-OH

DCCl/HOBt-coupling between N-(Cbz)-1-Piq-Pro-OH and H - Ppa(Boc)[CHOHCO] -OMe.HCl, saponification, oxidation dess - Martin, removing protection and purification were carried out by the methods described in Example 1. The target compound was obtained as a mixture of diastereoisomers.

Rt(LC): 19,8 and 20,31 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 14

H-D-Phe-Pro-Ppa[COCO]-

Boc-D-Phe-Pro-OH was obtained by a method similar to that described in Example 1. DCCl/HOBt-coupling between Boc-D-Phe-Pro-OH and H-Ppa(Boc)[CHOHCO]-OMe.HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1 and was obtained a mixture of diastereomers.

Rt(LC): 16,24 and 16,90 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 15

3,3-Diphenylpropionic-Pro-Ppa[COCO]-

3,3-Diphenylpropionic-Pro-OH was obtained by the method described in Example 1. DCCl/HOBt-coupling between 3,3-diphenylpropionic-Pro-HE and H-Ppa (Boc)[CHPHCO] -OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1 and was obtained a mixture of diastereomers.

Rt(LC): 36,60 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

TLC: Rf0,90, silica gel, ethyl acetate/heptane, 3/1, V/V.

(b) Boc-(N-Me)-D-Cha-OMe

Boc-D-Cha-OMe (19,35 g) was dissolved in 200 ml of dry N,N-dimethylformamide in a nitrogen atmosphere. After you have added under the conditions (4,22 ml) and the mixture was cooled to 0oC. Then was added a 2.71 g of sodium hydride (60% dispersion in oil) and the reaction mixture was stirred for 3 hours at room temperature. The mixture was partially concentrated, added ethyl acetate and the organic layer was washed with 0.1 M hydrochloric acid, water, 5% sodium bicarbonate solution and saturated saline, and then dried with sodium sulfate and concentrated. Output: 21,07 Boc-(N-Me)-D-Cha-Pro-OH was obtained by way described in Example 1. DCCl/HOBt-coupling between Boc-(N-Me)-D-Cha-Pro-OH and H-Ppa(Boc)[CHOHCO]-OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1. The target compound was isolated as a mixture of diastereoisomers.

Rt(LC): 23,14 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 17

N-(N-Me)-D-Phe-Pro-Ppa[COCO]-

Boc-(N-Me)-D-Phe-Pro-OH was obtained by the method described in Example 1. DCCl/HOBt-coupling between Boc-(N-Me)-D-Phe-Pro-OH and H-Ppa(Boc)[CHOHCO]-OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 16,26/16,90 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 18

SO2-D-Cha-Pro-Ppa[COCO]-

Boc-D-Cha-Pro-OPac (OPac=finacialy ester)

Boc-D-Cha-Pro-OPac was obtained by method similar to that described in Example 1 using Boc-D-Cha-OH and H-Pro-OPac.

TLC: Rf0.5 on silica, dichloromethane/methanol, 95:5, about/about.

(a) SO2-D-Cha-Pro-OPAC

Boc-D-Cha-Pro-OPac (3.8 g) was dissolved in TFA/dichloromethane (1/1) (about/on) (25 ml) and was stirred for 30 minutes at room temperature. The reaction mixture was evaporated in vacuum. Neocidin the triethylamine to maintain pH=8 throughout the reaction. The mixture was stirred for 3 hours at 0oC and was added 25 ml of water. After stirring for a further 30 minutes at room temperature the reaction mixture was concentrated in vacuum. The residue was dissolved in diethyl ether, washed with 1 n hydrochloric acid, water, 5% sodium bicarbonate solution and saturated saline, then was dried with sodium sulfate, filtered and evaporated in vacuum. After trituration of the crude substances with methanol received SO2-D-Cha-Pro-OPac (3.0 g).

TLC: Rf0,6; silica; dichloromethane/methanol=95/5, V/V.

(b) SO2-P-Cha-Pro-HE

To a solution of SO2-D-Cha-Pro-OPAC (10 g) in tetrahydrofuran (250 ml) was added 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran (84 ml). The reaction mixture was stirred for 30 minutes at room temperature and poured into water (1 liter). The aqueous solution was extracted with ethyl acetate. The combined organic layers are successively washed with 1 n hydrochloric acid and water, and then dried with sodium sulfate and concentrated in vacuum. The residue was purified by crystallization from ethyl acetate/diisopropyl ether to obtain SO2-D-Cha-Pro-HE (6.0 g).

(C) SO2-D-Cha-Pro-Ppa[COCO]-

DCCl/HOBt-coupling between SO2-D-Cha-Pro-HE and H - Ppa(Boc)[CHOHCO] -OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, to obtain the target compound as a mixture of diastereoisomers.

Rt(LC): 35,90 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 19

N-Me-D-Cha-Azt-Ppa-(2-thiazolyl)

(a) H-Azt-OBzl.HCl

N-Boc-Azt-OH (4,60 g) was dissolved in dichloromethane (50 ml) and was treated with benzyl alcohol (2,47 g), tetrafluoroborate 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea (7,40 g) and triethylamine (6.3 ml) and then stirred for 1 hour at room temperature. The reaction mixture was diluted with 1 n hydrochloric acid, after which the organic phase was separated and washed with water, 5% sodium bicarbonate and saturated saline solution. After filtration and evaporation in vacuum was obtained the crude product, which was purified by chromatography on silica gel using as eluent heptane/ethyl acetate =3/1, V/V, to obtain the N-Boc-Azt-OBzl (6.0 g). After removal of the protective group Boc in 3 M hydrochloric acid in dioxane (60 ml) for 1 hour at room temperature and after evaporation of the DOS the PTA/water, 63/20/6/11, about/about/about/about.

(b) Boc-(N-Me)-D-Cha-Azt-OH

1.5 g of Boc-(N-Me)-D-Cha-OH (see example 58) and 1.5 g of H-Azt-OBzl.HCl was subjected to interaction with the methods described in Example 1 for the synthesis of dipeptides. After the hydrogenolysis reaction (see Example 1) benzyl ether complex was obtained 1.22 g of Boc-(N - Me)-D-Cha-Azt-OH.

TLC: Rf0,50; silica gel; ethyl acetate/pyridine/acetic acid/water/ 63/20/6/11, about/about/about/about.

(C) N-Me-D-Cha-Azt-Ppa-(2-thiazolyl)

After binding assays Boc-(N-Me)-D-Cha-Azt-OH (370 mg) with H-Ppa(theos)-(2-thiazolyl) TsOH (610 mg) as described in Example 48, and after purification on silica gel in dichloromethane/methanol (95:5, V/V) was obtained Boc-(N-Me)-D-Cha-Azt-PPA(Teoc)-(2-thiazolyl) (684 mg). After removal of the protective groups (see Example 48) and after HPLC purification were obtained from two separate diastereoisomer.

Yield: 76 mg; Rt(LC): 26,9 min; 20%A/80% - 20%A/20%b and 60%C in 40 minutes.

Yield: 76 mg; Rt(LC): 30,3 min; 20%A/80% - 20%A/20%b and 60%C in 40 minutes.

Example 20

H-D-Hpl-Pro-Ppa[COCO]-

(a) H-D-Hpl-OMe

H-D-Cha-OH (1.0 g) was dissolved in a mixture of 1 n hydrochloric acid (4.8 ml), water (to 19.4 ml) and acetic acid (9.7 ml). At 0oC was slowly added a solution of sodium nitrite (3.4 g) in water (5.8 ml) and the mixture was stirred over night at room temperature. P temperature. The reaction mixture was concentrated, and the residue was dissolved in ether/acetone. After filtration the solution was concentrated in vacuum and the crude product was stirred in methanol (25 ml) for 18 hours. the pH was equal to 1.5. The reaction mixture was evaporated to dryness and the residue was purified by chromatography on silica (eluent: toluene/methanol=97/3, V/V) to give H-D-Hpl-OMe (612 mg).

TLC: Rf0,9; ethyl acetate/pyridine/acetic acid/water = 163/20/6/11, about/about/about/about, silicon dioxide.

(b) THP-D-Hpl-OMe (THP=tetrahydropyran)

To a stirred solution of H-D-Hpl-OMe (450 mg) in dichloromethane (2 ml), was added 3,4-dihydro-2H-Piran (0,29 ml) and p-toluensulfonate pyridinium (60 mg). The mixture was stirred for b hours at room temperature and was diluted with ether. This mixture was washed with saturated saline solution, dried with sodium sulfate, filtered and evaporated in vacuum. The crude product was purified by chromatography on silica (eluent: ethyl acetate/heptane, 1:4, V/V) to obtain the TPH-D-Hpl-OMe (498 mg).

TLC: Rf0,64; ethyl acetate/heptane=1:2, V/V on silica.

(C) THP-D-Hpl-OH

A solution of THP-D-Hpl-OMe (10.3 g) in dioxane/water (9/1, V/V) (200 ml) at room temperature for treatment is ect was poured into water (500 ml) and was extracted with dichloromethane. The organic layer was washed with water and dried with sodium sulfate. The filtrate was evaporated and obtained 6.6 g of the target compound.

TLC: Rf0,78; silica; ethyl acetate/pyridine/acetic acid/water = 163/20/6/11, about/about/about/about.

(d) THP-D-Hpl-Pro-OH

To a solution of THP-D-Hpl-OH (by 5.87 g) in acetonitrile (75 ml), was added EDCl (hydrochloride of 1-(3-dimethyl-aminopropyl)-3-carbodiimide (4,84 g) and N-hydroxysuccinimide (2.9 g). The reaction mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo, and the residue was dissolved in ethyl acetate. This solution was washed with water and saturated saline, and then dried with sodium sulfate and concentrated in vacuum. The crude product was dissolved in N,N-dimethylformamide (100 ml) was added to a solution of Proline hydrochloride (6,99 g) in N,N-dimethylformamide/water, 1/1, V/V (200 ml), which was brought to pH of 8.5 by adding sodium hydroxide. After stirring over night the reaction mixture was concentrated in vacuo and the residue was dissolved in water. This aqueous solution was brought to pH=2,5 at 0oC, and then were extracted with ethyl acetate. The combined organic layers are then washed with water and saturated saline, and then the silica (eluent: ethyl acetate/methanol=8/2 ---> 6/4, V/V) and received THP-D-Hpl-Pro-OH (6.75 g)

TCX: Rf0,52; ethyl acetate/pyridine/acetic acid/water= 163/20/6/11, about/about/about/about, silicon dioxide.

(e) H-D-Hpl-Pro-Ppa[COCO]-OH

DCCl/HOBt-coupling between THp-D-Hpl-Pro-OH and H-Ppa(Boc)[CHOHCO]-OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained a mixture of diastereomers.

Rt(LC): 31,05 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 21

3,3-Diphenylpropionic-Pro-Ppa[COCO]-OMe

3,3-Diphenylpropionic-Pro-HE was obtained by the methods described in Example 1. DCCl/HOBt-coupling between 3,3-diphenylpropionic-Pro-HE and H-Ppa(Boc)[CHOHCO] -OMe. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 21,7 min; 20%A/60%/ 20% - 20%A/80%C for 30 minutes.

Example 22

SO2-D-Cha-Pro-Ppa[COCO]-

SO2-D-Cha-Pro-HE described in Example 18. DCCl/HOBt-coupling between SO2-D-Cha-Pro-OH and H-Ppa(Boc)[CHOHCO]-OMe.HCl oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(Rockstarmarketer)-N-Boc-D-Cha-Pro-OH as described in Example 1. DCCl/HOBt-coupling between N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-HE and H-Ppa(Boc)[CHOHCO]-OMe.HCl oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained a mixture of diastereomers.

Rt(LC): 25,75 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 24

SO2-D-Cha-Pro-Ppa[COCO]-OEt

SO2-O-Cha-Pro-OH was obtained by methods described in Example 18.

(a) Cbz-Ppa[CHOHCO]-OEt

A solution of Cbz-Ppa(Boc)[CHOHCO] -OMe (1 g) in 50 ml of 3 M hydrogen chloride/ethanol at -20oC was stirred for 4 hours, raising the temperature to room temperature. The mixture was evaporated in vacuum and obtained 0.9 g of target compound in the form of butter.

TLC: Rf0,85; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(b) Cbz-Ppa(Boc)[CHOHCO]-OEt

the pH of the solution of 0.9 g of Cbz-Ppa[CHOHCO]-OEt and 0.55 g of di-t-BUTYLCARBAMATE in 10 ml of N,N-dimethylformamide was brought to pH 8.5 by the addition of triethylamine. The resulting mixture was stirred for 1 hour at room temperature, poured into water and was extracted with ethyl acetate. The combined extracts washed with 1 M hydrochloric acid, water, 5% sodium bicarbonate solution and water and then dried with sodium sulfate and evaporated in vacuum with P1, about/about).

C) SO2-D-Cha-Pro-Ppa[COCO]-OEt

Cbz-Ppa(Boc)[CHOHCO]-OEt was first made by the method described in Example 2(q) and received H-Ppa(Boc)[CHOHCO]-OEt.HCl.

DCCl/HOBt-coupling between SO2-O-Cha-Pro-OH and H-Ppa (Boc)[CHOHCO] -OEt. HCl, saponification, oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, resulting in the obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 41,58 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 25

HOOC-CH2-D-Cha-Pro-Ppa[COCO]-OEt

N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-HE described in Example 1. DCCl/HOBt-coupling between N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-HE and H-Ppa(Boc)[CHOHCO]-OEt.HCl oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 28,20 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 26

SO2-D-Phe-Pro-Ppa[COCO]-OEt

SO2-D-Phe-Pro-HE was obtained by the methods described in Example 18. DCCl/HOBt-coupling between SO2-D-Phe-Pro-HE and H-Ppa(Boc)[CHOHCO]-OEt. HCl oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained a mixture of diastereomers.D-Cha-Pro-Ppa(Eoc)[COCO]-OEt

(Eoc=etoxycarbonyl)

(a) SO2-D-Cha-Pro-Ppa[CHOHCO]-OEt

SO2-D-Cha-Pro-Ppa (Boc) [CHOHCO]-OEt was obtained by the method described in Example 24. Complex hydroxyethyloxy ester (200 mg) was treated triperoxonane acid /dichloromethane (1/1) for 1 hour at room temperature to obtain 210 mg of the target compound.

TLC: Rf0,85; silica gel; ethyl acetate/pyridine/acetic acid/water, 63/20/6/11, about/about/about/about.

(b) SO2-D-Cha-Pro-Ppa(Eoc)[CHOHCO]-OEt

the pH of the solution of 210 mg SO2-D-Cha-Pro-Ppa[CHOHCO]-Et and 43.6 ál ethylchloride in 10 ml of N,N-dimethylformamide was brought to pH 8.5 by the addition of triethylamine. The mixture was stirred for 1 hour at room temperature, poured into water and was extracted with ethyl acetate. The combined extracts washed with 1 M hydrochloric acid, water, 5% sodium bicarbonate solution and again with water, and then dried with sodium sulfate and evaporated in vacuum to obtain 220 mg of the target compound in the form of butter.

TLC: Rf0,50; silica gel; ethyl acetate/pyridine/acetic acid/water= 463/20/6/11, about/about/about/about.

(C) SO2-D-Cha-Pro-Ppa(Eoc)[COCO]-OEt

Oxidation dessa-Martin and purification was performed as described in Example 1, and the received target soedinenii 28

SO2-D-Cha-N-cyclopentyl-Gly-S-Ppa-(2-thiazolyl)

(a) SO2-D-Cha-N-cyclopentyl-Gly-HE

Boc-D-Cha-OH (2.65 g) was subjected to interaction with the N-cyclopentyl-Gly-OMe. HCl (Cm. Example 2), as described in the method dipeptide synthesis in Example 1 and after purification on silica gel was obtained N-Boc-D-Cha-N-cyclopentyl-Gly-OMe (3.5 g). After removal of the protective group Boc in TFA/dichloromethane (1/1, V/V) (38 ml) and after reaction metilsulfonilmetane as described in Example 18 was obtained SO2-D-Cha-N-cyclopentyl-

Gly-OMe (1.4 g). The hydrolysis of complex methyl ester was carried out by the methods described in Example 2, and received SO2-D-Cha-N-cyclopentyl-Gly-HE.

TLC: Rf0,2; silica gel; dichloromethane/methanol=9/1, V/V.

(b) (2S)-2-Azido-3-(piperidine-4-yl)-propanoic acid

A solution of 3.7 g of (2S)-2-azido-3-[1-(t-butyloxycarbonyl)-piperidine-4-yl]-propanoic acid (Example 12(e)) was treated with 30 ml of 3 M hydrogen chloride in ethyl acetate. The mixture was stirred for 30 minutes at room temperature, evaporated in vacuum and received 3.0 g of crystalline solids.

TLC: Rf0,35 on silica; ethyl acetate/pyridine/acetic acid/water = 63/20/6/11, about/about/about/about.

(C) (2S)-2-Azido-3-[1-[(2-trimethylsilyl) ataxia-(trimethylsilyl) ethoxycarbonyl] pyrrolidin-2,5-dione (TeocOSu, link: Synthesis 1987, 346) brought to 8.5 using triethylamine. The reaction mixture was stirred for 30 minutes at room temperature, poured into water and was extracted with ethyl acetate. The combined extracts washed with 1 n hydrochloric acid, 5% sodium bicarbonate and water and then dried with sodium sulfate. In the evaporation in vacuum received is 4.93 g of the desired compound in the form of butter.

TCX: Rf0,85, ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about, silicon dioxide.

(d) (2S)-N-Methyl-N-methoxy-2-azido-3-[1-[(2-trimethylsilyl)etoxycarbonyl]piperidine-4-yl]-propanamide

(2S)-N-Methyl-N-methoxy-2-azido-3-[1-[(2-trimethylsilyl)- etoxycarbonyl] piperidine-4-yl] -propanamide was obtained by the method described in Example 12(f), using (2S)-2-Azido-3-[1-[(2-trimethylsilyl) etoxycarbonyl] piperidine-4-yl]-propanoic acid.

TCX: Rf0,85: silica gel; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(e) H-S-Ppa(Teoc)-N(Me)OMe

Hydrogen was passed through a mixture of 4.9 g of (2S)-N-Methyl-N-methoxy-2-azido-3-[1-[(2-trimethylsilyl)etoxycarbonyl] -piperidine-4-yl] -propanamide and 0.5 g of Pd/C (10%) in 100 ml of tetrahydrofuran and 13.3 ml of 1 n hydrochloric acid until then, until the entire IP 5.7 g of the desired compound.

TCX: Rf0.9, ethyl acetate/pyridine/acetic acid/water = 63/20/6/11, (about/about/about/about) on silicon dioxide.

(f) Boc-S-Ppa(Teoc)-N(Me)OMe

the pH of the solution of 4.7 g H-S -- Ppa(Teoc)-N(Me)OMe and 2.8 g of di-t-BUTYLCARBAMATE brought to pH = 8,5 using triethylamine. The mixture was stirred for 1 hour at room temperature, poured into water and was extracted with ethyl acetate. The combined organic layers washed with 1 n hydrochloric acid, water, 5% sodium bicarbonate and again with water, and then dried with sodium sulfate and evaporated in vacuum to obtain 5.2 g of the target compound in the form of butter.

TLC: Rf0,85, ethyl acetate/pyridine/acetic acid/water = 63/20/6/11, (about/about/about/about) on silicon dioxide.

Chiral HPLC on a column of Chiralpack Chir 11 h,6 in hexane/ethanol (9/1, V/V) gave 91% enantiomeric excess.

(g) BOC-3-PPA(Teoc)-(2-thiazolyl)

To a solution of 31.7 ml of 1.05 M n-BuLi in ether at -78oC in an atmosphere of nitrogen was added 5,46 ml of 2-bromothiazole. Received pre-cooled yellow solution (2.5 EQ.) the cannula was added to a solution of Boc-S-Ppa(Teoc)-N(Me)OMe in 46 ml of dry tetrahydrofuran at -78oC in nitrogen atmosphere. The reaction mixture was stirred for 30 minutes at -78oC in nitrogen atmosphere, howled the sewed sodium sulfate and evaporated in vacuum to obtain 5.0 g of oily residue. After flash chromatography in heptane/ethyl acetate (7/3, V/V) on silica was obtained 1.7 g of the target compound.

TLC: Rf0.75, heptane/ethyl acetate, 7/3, V/V on silica.

[]D= +31,9o(C=0.45 in CHCl3)

(h) N-3-PPA(Teoc)-(2-thiazolyl).TsOH

To a solution of 1.54 g of the BOC-3-PPA(Teoc)-(2-thiazolyl) to 14.2 ml of dry ether was added a solution of 0.96 g of monohydrate p-toluensulfonate acid, 2.8 ml of dry ethanol. The solution was kept for 1 hour at 30oC and the ether was removed. The obtained solid was washed with dry ether and was obtained 1.5 g of target compound in the form of a solid substance.

TLC: Rf0,85, dichloromethane/methanol =9:1, about/about.

(i) SO2-D-Cha-N-cyclopentyl-Gly-3-Ppa-(2-thiazolyl)

Linking SO2-D-Cha-N-cyclopentyl-Gly-HE (395 mg) with H-S-Ppa(Teoc)-(2-thiazolyl). TsOH was carried out by the methods described in Example 48, and received 400 mg fully protected Tripeptide. The Teoc group was removed (see Example 48) and received 300 mg of the crude SO2-D-Cha-N-cyclopentyl-Gly-S-Ppa-(2-thiazolyl). TFA. This salt (150 mg) was purified by HPLC and received 112 mg of the target compound.

Rt(LC): 32,06 min; 20%A, 60%b and 20%C - 100%C for 40 minutes.

Example 29

SO2 was dissolved in N,N-dimethylformamide (10 ml), handled etelcharge.com (38 ml) at a pH of about 8.5 (using triethylamine). And then was stirred for 2 hours at room temperature. The reaction mixture was diluted with ethyl acetate (100 ml) and then washed with 1 n hydrochloric acid, water, 5% sodium bicarbonate and again with water. The organic phase was dried with sodium sulfate, filtered and evaporated to dryness to obtain SO2-D-Cha-N-cyclopentyl-Gly-S-Ppa(Eoc)-(2-thiazolyl).

TCX: Rf0,45; silica; dichloromethane/methanol, 9/1, V/V.

Example 30

HOOC-CH2-D-Cha-Pro-Ppa[COCO]-NH2< / BR>
(a) N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[CHOHCO]-OH

N-(t-Butyloxycarbonyl)-N-BOC-D-Cha-Pro-OH as described in Example 1.

DCCl/HOBt-coupling between N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-OH and H-Ppa(Boc)[CHOHCO] -OMe. HCl and saponification was carried out by the method described in Example 1.

TCX: Rf0,80: silica gel; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(b) N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro - Ppa (Boc)[CHOHCO]-NH2< / BR>
N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[CHOHCO]-OH was dissolved in N, N-dimethylformamide (12 ml). The solution was cooled in a bath of ice and water. And then added for 91.3 mg HOBT, 91,3 μl of N-methylmorpholine, and 50.5 mg Alibali in 1 M hydrochloric acid and was extracted with ethyl acetate. The combined extracts were washed with water, 5% sodium bicarbonate solution and water and then dried with sodium sulfate and evaporated in vacuum to obtain 400 mg of the target compound.

TCX: Rf0,25: silica gel; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(C) HOOC-CH2-D-Cha-Pro-Ppa[COCO]-NH2< / BR>
Oxidation dess-Martin, removing protection and purification was performed as described in Example 1. Output: a 94.2 mg of target compound as a mixture of diastereoisomers.

Rt(LC): 22,97 min;20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 31

HOOC-CH2-D-Cha-Pro-Ppa[COCO]-NHMe

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[CHOHCO] -NHMe was obtained by the method described in Example 30, using methylamine. Oxidation dess-Martin, removing protection and purification was performed as described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 24.22 to and 24,99 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 32

HOOC-CH2-D-Cha-Pro-Ppa[COCO]-(1-azetidin)

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[CHOHCO]-(1-azetidin) was obtained by the method described in Example 30, using the hydrochloride of azetidine. Oxidation dess-Martin, removing sewn and PTS to 28.05 and of 28.34 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 33

HOOC-CH2-D-Cha-Pro-Ppa[COCO]-(NH-(CH2)2-phenyl:

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[CHOHCO] -(NH-(CH2)2-phenyl) received by the method described in Example 30, using 2-(phenyl)ethylamine. Oxidation dess-Martin, removing protection and purification were carried out by the method described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 38,82 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 34

HOOC-CH2-D-Cha-Pro-Ppa[COCO]-(NH-CH2phenyl)

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Boc)[CHOHCO] -(NH-CH2phenyl) was obtained by the method described in Example 30, using 2-(phenyl)ethylamine. Oxidation dess-Martin, removing protection and purification were carried out by the method described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 36,46 min, 20%A/80% - 20%A/20%/60%for 4 0 minutes.

Example 35

SO2-D-Cha-Pro-Ppa[COCO]-NH2< / BR>
SO2-D-Cha-Pro-HE described in Example 18. DCCl/HOBt-coupling between SO2-D-Cha-Pro-OH and H-Ppa(Boc)[CHOHCO] -OMe. HCl, saponification and response interaction EDCl with ammonium chloride was carried out as described in Example 30. the camping diastereomers.

Rt(LC): 36,70 min; 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 36

SO2-D-Cha-Pro-Ppa[COCO]-(NH-CH2phenyl)

SO2-D-Cha-Pro-OH was obtained as described in Example 18. DCCl/HOBt-coupling between SO2-D-Cha-Pro-HE and H-Ppa(Boc)[CHOHCO]-OMe.HCl, saponification and response interaction EDCl with benzylamine carried out by the methods described in Example 30. Oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, to obtain the target compound as a mixture of diastereoisomers.

Rt(LC): 28,76 and 29.1 min, 20%A/60%B/20%C - 20%A/80%within 30 minutes.

Example 37

SO2-D-Cha-Pro-Ppa[COCO]-Ne2< / BR>
SO2-D-Cha-Pro-OH was obtained as described in Example 18. DCCl/HOBt-coupling between SO2-D-Cha-Pro-HE and H - Ppa(Boc)[CHOHCO]-OMe.HCl, saponification and response interaction EDCl with dimethylamine were carried out by the methods described in Example 30. Oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained target compound as two diastereomers.

A: Rt(LC): 41,96 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

IN: Rt(LC): 43,49 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 38

SO2- D-Phe-Pro-OH and H-Ppa(Boc)[CHOHCO]-OMe.HCl, saponification and response interaction EDCl with ammonium chloride were carried out using the methods described in Example 30. Oxidation dess-Martin, removing protection and purification was performed as described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 28,81 and 29,25 min, 20%A/80%-20%A/20%/60%C in 40 minutes.

Example 39

HOOC-CH2-p-Cl-D-Phe-Pro-Ppa[COCO]-NH2< / BR>
N-(t-Butyloxycarbonyl)-N-Boc-p-Cl-D-Phe-Pro-OH was obtained as described in Example 5. DCCl/HOBt-coupling between N-(t-Butyloxycarbonyl)-N-BOC-p-Cl-D-Phe-Pro-HE and H-Ppa(Boc)[CHOHCO]-OMe.HCl, saponification and response interaction EDCl and ammonium chloride were carried out using the methods described in Example 30. Oxidation dess-Martin, removing protection and purification were carried out by the methods described in Example 1, and obtained target compound in the form of a mixture of diastereoisomers.

Rt(LC): 23,35 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 40

HOOC-CH2-p-Cl-D-Cha-Pro-Ppa[COCO]-(NH-CH2phenyl)

N-(t-Butyloxycarbonyl)-N-Boc-p-Cl-D-Phe-Pro-OH was obtained as described in Example 5. DCCl/HOBt-coupling between N-(t-Butyloxycarbonyl)-N-BOC-p-Cl-D-Phe-Pro-HE and H - Ppa(Boc)[CHOHCO]-OMe.HCl, saponification and reaction of co who you and purification was performed by methods, described in Example 1, and received two diastereoisomer.

A: Rt(LC): 19,29 min, 20%A/60%/20% - 20%A/80%within 30 minutes.

IN: Rt(LC): 37,61 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 41

[1-Oxo-2-[3-(phenylmethylsulfonyl)-2-oxo-2H-pyridin-1-yl] ethyl] -Ppa[COCO]-(NH-CH2phenyl)

(a) [3-(Phenylmethylsulfonyl)-2-oxo-2H-pyridin-1-yl]-acetic acid

(3-Amino-2-oxo-2H-pyridin-1-yl)-acetic acid ethyl ester described in J. Med. Chem. , 1996, 37, 3090-3099. Subsequent reaction of sulfonylurea and saponification was performed by the method described in Example 12.

TLC: Rfof 0.4, silica gel; ethyl acetate/pyridine/acetic acid /water= 63/20/6/11, about/about/about/about.

(b) SO2-3-NH2-1-carboxymethylamino-2-on-PPA[COCO]-(NH-CH2phenyl)

Linking between SO2-3-NH2-1-carboxymethylamino-2-one and H-Ppa(Boc)[CHOHCO]-OMe.HCl (see Example 1), saponification, amide linking, oxidation dess-Martin, release and purification was performed as described in Example 34, and received two diastereoisomer a and B.

A: Rt(LC): 21,6 min, 20%A/60%b and 20%C - 100%C for 40 minutes.

IN: Rt(LC): 22,8 min, 20%A/60%b and 20%C - 100%C for 40 minutes.

Example 42

Bunshichi described in Example 12. DCCl/HOBt-coupling between SO2-norLey(cyclo)Gly-HE H-S -- Ppa(Boc)(CHOHCO] -OMe. HCl (Example 12(m)), saponification and response interaction EDCl with benzylamine carried out by the methods described in Example 30. Oxidation dess-Martin, removing protection and purification was performed as described in Example 1.

Rt(LC): 23,46 min, 20%A/60%/20% - 20%A/80%within 30 minutes.

Example 43

HOOC-CH2-D-Cha-Pro-Ppa-(2-benzothiazolyl)

(a) Boc-Ppa(Cbz)-OMe

Boc-Ppa(Cbz)-OMe was obtained from Boc-Ppa(Cbz)-OH (example 45) in the manner similar to that described in Example (m).

(b) Boc-Ppa(Cbz)-(2-benzothiazolyl)

To a cold (-30oC) and stirred solution of n-BuLi in hexane (3.9 ml, 1.3 M) was added dropwise a solution of benzothiazole (675 mg) in tetrahydrofuran (25 ml). The solution was stirred for 15 minutes at -30oC, and then was slowly added to a solution of Boc-Ppa(Cbz)-OMe (1.0 g) in dry tetrahydrofuran (25 ml) at - 40oC. This mixture was stirred for 2 hours at -20oC, and then added a 5% aqueous sodium bicarbonate solution. The mixture was left to warm to room temperature and the layers were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, dried with sodium sulfate, filtered and evaporated to obtain and have obtained target compound (230 mg).

TLC: Rfof 0.35, silica gel; heptane/ethyl acetate=1/2, about/about.

(C) N-(t-Butyloxycarbonyl) (Boc)-D-Cha-Pro-Ppa(Cbz)-(2-benzothiazolyl)

Release Boc-Ppa(Cbz)-(2-benzothiazole) and linking with N-(t-butyloxycarbonyl)(Boc)-D-Cha-Pro-OH were carried out by the method described in Example 45. After purification by chromatography on silica gel using gradient elution heptane/ethyl acetate (1/1, V/V) ---> heptane/ethyl acetate (1/2, V/V) were given 200 mg of the target compound.

TLC: Rfof 0.5, silica gel; heptane/ethyl acetate=1/2, about/about.

(d) HOOC-CH2-D-Cha-Pro-Ppa-(2-benzothiazolyl)

To a mixture of 6 ml triperoxonane acid and 0.6 ml of thioanisole was added at room temperature, 0.45 g of N-(t-butyloxycarbonyl)(Boc)-D-Cha-Pro-Ppa(Cbz)-(2-benzothiazolyl). After 4 hours the reaction mixture was concentrated, and the residue was dissolved in water. The aqueous phase was washed with diethyl ether and concentrated in vacuum. The crude product was purified on a preparative HPLC column (C18, 15 μm, Deltapack, 100A using a gradient elution of 20% A/80% - 20% A/35%/45% for 45 minutes at a flow rate of 50 ml/min and got the target compound in the form of two diastereoisomers.

Output: 197 mg, Rt(LC): 38,70 min, 20%A/80% is.

Example 44

HOOC-CH2-D-Cha-Pro-Ppa-(2-oxazolyl)

(a) 1-[[2-(Trimethylsilyl)ethoxy] carbonyl] -4-[2-(t-butyloxycarbonyl)-3-hydroxy-3-(oxazol-2-yl)propyl]piperidine

To a solution of 3.03 g of Boc-Ppa(Teoc)N(Me)OMe (example 48) in 50 ml of dichloromethane at -78oC in an atmosphere of nitrogen was added 20 ml of 1 M solution of hydride diisobutylaluminum in hexane. After one hour the cooling bath was removed, and then immediately added to 100 ml of 0.25 M hydrochloric acid and an additional amount of dichloromethane and after 5 minutes the resulting suspension was filtered. To the filtrate was added a saturated saline solution, then the organic layer was separated, dried (sodium sulfate) and concentrated to obtain a 2.36 g of Boc - Ppa(Teoc)-H in the form of oil. This aldehyde (2,36 g) and 1.86 g of 2-(trimethylsilyl) oxazole (Edwards, P. D., Wolanin, D. J., Andisik D. W. and W. Davis, J. Med.Chem., 38, 76 (1995)) were mixed and heated at 50oC for 3 hours. Then the reaction mixture was cooled to room temperature and after three days this reaction mixture was concentrated. The residue was dissolved in 10 ml of tetrahydrofuran and 1 ml of 1 n hydrochloric acid. After incubation for 1 hour at room temperature to the solution was added ethyl acetate and a saturated saline solution, and the organic layer was separated and concentrated. The hydrolysis reaction was not completed, and the residue was again dissolved in 10 ml of tetrahydrofuran and 1 ml of 1 n hydrochloric acid. After incubation for 1 hour at room temperature was added ethyl acetate and a saturated saline solution and then the organic layer was separated. The aqueous layer three times were extracted with ethyl acetate and the combined organic layers were dried (sodium sulfate) and concentrated. After purification using column chromatography on silica gel (eluent: ethyl acetate/heptane, 3/2, V/V) was obtained 0,70 g of target compound.

TLC: Rf of 0.2, silica gel; ethyl acetate/heptane, 1/1, V/V.

(b) Boc-Ppa(theos)-(2-oxazolyl)

To a solution 0,70 g of 1-[[2-(trimethylsilyl)ethoxy]carbonyl]-4-[2-(t-butyloxycarbonyl)-3-hydroxy-3-(oxazol-2-yl)propyl] piperidine in 10 ml of dichloromethane was added 0.6 g of reagent dess-Martin. After stirring for one hour at room temperature was added 50 ml of 5% aqueous sodium thiosulfate solution and the mixture was stirred for 45 minutes at room temperature. The organic layer was separated, washed with water, 5% aqueous sodium bicarbonate solution and saturated saline, and then dried with sodium sulfate and concentrated. After ochistit.

TLC: Rfof 0.5, silica gel; heptane/ethyl acetate = 1/1, V/V.

(C) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-PPA(Teoc)-(2-oxazolyl)

To a solution of 477 mg of Boc-Ppa(Teoc)-(2-oxazolyl) in 12 ml of diethyl ether and 2 ml of ethanol was added 213 mg of the monohydrate para-toluensulfonate acid and the ether was removed under reduced pressure at 30oC. the Oily residue was heated at 60oC for 20 minutes, and then the ethanol was removed under reduced pressure and the residue was dried in vacuum to obtain 0.5 g H-Ppa(Teoc)-(2-oxazolyl). TsOH. To a solution of 0.29 g of N-(butyloxycarbonyl)-N-BOC-D-Cha-Pro-HE (described in Example 1) in 10 ml of N,N-dimethylformamide in a nitrogen atmosphere at -20oC was added 0,078 ml isobutylphthalate and 0,208 ml of N,N-diisopropylethylamine. After 30 minutes the solution was added 0.25 g of H-Ppa(Teoc)-(2-oxazolyl). TsOH in N,N-dimethylformamide and 0,050 ml of N,N-diisopropylethylamine and the reaction mixture was heated to room temperature. After two hours the reaction mixture was concentrated under reduced pressure. The residue was dissolved in ethyl acetate, washed with 5% sodium bicarbonate solution and saline. And then was dried (sodium sulfate) and concentrated. After purification by chromatography on silica gel using a gradient elwira the

TLC: Rfof 0.5, silica gel; heptane/ethyl acetate = 1:2, about/about.

(d) HOOC-CH2-D-Cha-Pro-Ppa-(2-oxazolyl)

To a solution of 0.20 g of N-(t-butyloxycarbonyl)(Boc)-D-Cha-Pro-Ppa(Teoc)-(2-oxazolyl) in 2 ml dichloromethane was added 2 ml triperoxonane acid and the mixture was stirred at room temperature. After 4 hours the reaction mixture was concentrated and was purified on preparative HPLC column C18 15 μm 100A Deltapack using a gradient elution of 20% A/80%/0% - 20%/51%/29%for 45 minutes at a flow rate of 20 ml/min, to obtain the target compound as two diastereomers.

Yield: 49 mg Rt(LC): 27,83 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Yield: 46 mg Rt(LC): 30,04 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 45

HOOC-CH2-D-Cha-Pro-Ppa-(2-thiazolyl)

(a) Boc-Ppa(Cbz)-OH

the pH of a solution of 70 g of H-Ppa-OH, 39,85 g of the pentahydrate of copper sulfate (II) and 147 g of benzyloxycarbonylglycine brought to 9 with 2 n sodium hydroxide. The mixture was stirred for 16 hours at room temperature. The precipitate was collected and thoroughly washed with water. The filter cake was dissolved in dioxane, and the pH was brought up to 12.5 using 4 n sodium hydroxide, and then added 129 g of di-t who has koncentrirebuli to small volume and the pH was brought to 2.5. The residue was diluted with ethyl acetate. The organic layer was washed with water, dried with sodium sulfate and was evaporated to dryness to obtain 100 g of oil.

TLC: Rf0,77, dichloromethane/methanol (9/1, V/V) on silica gel.

(b) Boc-Ppa(Cbz)-N(Me)(OMe)

To a solution of 22.5 g of Boc-Ppa(Cbz)-OH in 500 ml dichloromethane was added rate of 7.54 g of the hydrochloride of N, O-dimethylhydroxylamine and 24,75 g TBTU. the pH was brought to 8.5 by the addition of triethylamine. The mixture was stirred for 2 hours at room temperature, and then washed with 1 n hydrochloric acid, water, 5% sodium bicarbonate solution and again with water, then dried with sodium sulfate and was evaporated in vacuo, resulting in received of 15.4 g of white solid product after crystallization from diisopropyl ether.

TCX: Rfof 0.5, dichloromethane/ethyl acetate = 8/2, V/V on silica.

(C) Boc-Ppa(Cbz)-(2-thiazolyl)

To a cold (-78oC) stirred solution of n-BuLi (0.11 mol) in diethyl ether (100 ml) was added dropwise a solution of 2-bromothiazole (18.2 g, 0.11 mol) in diethyl ether (100 ml). After stirring the solution at -78oC for 30 minutes and thereto was slowly added a solution of Boc-Ppa(Cbz)-N(Me) (OMe) (15 g, 0.035 mol) in dry tetrahydrofuran (300 ml). Blend the Mixture was heated to room temperature and the layers were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water, dried with sodium sulfate, filtered and evaporated to obtain 15.1 g of oil. The residue was purified by chromatography on silica (eluent: dichloromethane/methanol, 99/1, V/V) and was obtained 3.2 g of the target compound.

TCX: Rfof 0.45, silica gel, dichloromethane/methanol, 95/5, about/about.

(d) H-Ppa(Cbz)-(2-thiazolyl).TFA

N-Boc-Ppa(Cbz)-(2-thiazolyl) (500 mg) was dissolved in 10 ml triperoxonane acid/dichloromethane (1:1, V/V) and stirred for 1 hour at room temperature. The crude amine was isolated as an oil in quantitative yield after removal of the solvent by evaporation and used directly to obtain N-(t - butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Cbz)-(2-thiazolyl).

TLC: Rfof 0.4, silica gel; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(e) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Cbz)-(2-thiazolyl)

Isobutylparaben (130 μl) was added to a stirred solution of N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-HE (490 mg) and N,N-diisopropylethylamine (170 μl) in dry N,N-dimethylformamide (12 ml) at -15oC in nitrogen atmosphere. After 15 minutes to a cold solution smecheria at this pH to 8.5 by adding N,N-diisopropylethylamine. The reaction mixture was stirred for 60 minutes at -15oC. the Reaction mixture was evaporated to dryness. The residue was dissolved in ethyl acetate, successively washed with 2% aqueous citric acid solution, water and saturated saline, and then dried with sodium sulfate and concentrated in vacuum. The residue was purified by chromatography on silica (eluent: dichloromethane/methanol, 99/1, V/V) to give 470 mg of N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa (Cbz)-(2-thiazolyl).

TLC: Rf0,65, silica gel, dichloromethane/methanol=95/5, V/V.

(f) HOOC-CH2-D-Cha-Pro-Ppa-(2-thiazolyl)

A solution of 470 mg of N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro-Ppa(Cbz)-(2-thiazolyl) in 5 ml triperoxonane acid and 0.5 ml of thioanisole was stirred for 4 hours at room temperature. The solvent was removed in vacuo and the residue was purified by preparative RP-HPLC-column 18 Delta-Pak system using gradient elution: 20%: 70%: 10%: 20%: 20%: 60% for 40 minutes at a flow rate of 80 ml/min

(A: 0.5 M phosphate buffer, pH=2,1; B: water: acetonitrile:water, 3:2, V/V).

Yield: 177 mg Rf(LC): 33,57 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 46

HOOC-CH2-p-MeO-D-he-Rgvia H-Ppa(Cbz)-(2-thiazolyl) (135 mg), followed by the removal of protection and purification was performed by way described in Example 45, and got two single diastereoisomer.

Yield: 37 mg Rt: 27,17 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Yield: 47 mg Rt: to 30.15 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Example 47

HOOC-CH2-p-Cl-D-Phe-Pro-Ppa-(2-thiazolyl)

Synthesis of N-(t-butyloxycarbonyl)-N-Boc-p-Cl-D-Phe-Pro-IT is described in Example 5. The reaction of interaction with H-Ppa(Cbz)- (2-thiazolyl) (322 mg), followed by the removal of protection and purification was performed by the method described in Example 45, and got two single diastereoisomer.

Yield: 73 mg Rt: 31,07 min, 20%A/60%/20% - 20%A/0%/80%C for 30 minutes.

Yield: 94 mg Rt: 18,21 min, 20%A/80%/20% - 20%A/0%/80%C for 30 minutes.

Example 48

HOOC-CH2-D-Nle-Pro-Ppa-(2-thiazolyl)

Synthesis of N-(t-butyloxycarbonyl)-N-Boc-D-Nie-Pro-OH as described in Example 8.

(a) Boc-Ppa-N(Me)OMe

To a solution of Boc-Ppa(Cbz)-N(Me)OMe (Example 45b) (820 mg) in N,N-dimethylformamide (25 ml) was added 2 n hydrochloric acid (0,915 ml) and 10% palladium-on-coal (100 mg), and then the mixture was first made for 1 hour at room temperature. The reaction mixture was filtered through Diezel, washed with N, N-dimethylformamide and was neutralized with triethylamine (0.15 ml). The product was used crude is alali 1-[2-(trimethylsilyl)ethoxycarbonyl] pyrrolidin-2,5-dione (TeocOSu, link: Synthesis 1987, 346) (567 mg) and triethylamine (0,350 ml), and then the mixture was stirred for 4.5 hours at room temperature in a nitrogen atmosphere. The solution was maintained at pH= 8 with triethylamine. The solvent was removed by evaporation under reduced pressure, and then was added dichloromethane and the solution was washed with 0.1 n hydrochloric acid (100 ml), water, 5% sodium bicarbonate solution and water, then dried with magnesium sulfate and concentrated in vacuum. The residue was purified by chromatography on silica gel using as eluent heptane/ethyl acetate, 8/2, about/about. Output: 784 mg

TLC: Rf0,37, silica gel, heptane/ethyl acetate=1/1, V/V.

(C) Boc-Ppa(Teoc)-(2-thiazolyl)

To a cold (-70oC) a solution of n-utility in diethyl ether (68,4 ml, 1.07 mol/l) under nitrogen atmosphere was slowly added a solution of 2-bromothiazole (6,59 ml) in diethyl ether (50 ml) and the mixture was stirred for 30 minutes at -70oC. This solution thiazolide lithium is very slowly added to a cold (-70oC) a solution of Boc-Ppa(Teoc)-N(Me)OMe (16,8 g) in dry tetrahydrofuran (150 ml) and then was stirred for 1 hour at -70oC in nitrogen atmosphere. The reaction mixture was diluted with 0.2 n hydrochloric acid is Nata sodium and again with water, then were dried with magnesium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel using as eluent heptane/ethyl acetate, 8/2, about/about. Output: 11,35,

TLC: Rf 0,68, silica gel, heptane/ethyl acetate=1:1, about/about.

(d) H-Ppa(Teoc)-(2-thiazolyl).TsOH

To a solution of Boc-Ppa(Teoc)-(2-thiazolyl) (2.5 g) in diethyl ether (100 ml) was added p-toluensulfonate acid (1.2 g). The reaction mixture was slowly evaporated at 30oC at 200 mbar and for 1.5 hours at 15 mbar. Output: 3,03,

(e) HOOC-CHz-D-Nle-Pro-Ppa-(2-thiazolyl)

Binding of N-(t-butyloxycarbonyl)-N-Boc-D-Nle-Pro-HE H-Ppa(Teoc)-(2-thiazolyl).TsOH (307 mg), followed by the removal of protection and purification was performed by the method described in Example 45, and got two single diastereoisomer.

Yield: 96 mg Rt: 23,45 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Output: 99,6 mg Rt: 26,60 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Example 49

HOOC-CH2-m-Cl-D,L-Phe-Pro-PPA-(2-thiazolyl)

Synthesis of N-(t-Butyloxycarbonyl)-N-BOC-m-Cl-D, L-Phe-Pro-OH is described in Example 4. The binding of H-Ppa(Teoc)-(2-thiazolyl).TsOH (325 mg), followed by the removal of protection and purification was performed by the method described in Example 48, and received four separate diastereomer the t: 31,02 min, 20/A/80%/0% - 20%A/20%/60%C in 40 minutes.

Yield: 72 mg Rt: 32,62 min, 20%A/80%/60% C to 20%A/20%/60% C in 40 minutes.

Yield: 52 mg Rt: 34,22 min, 20%A/80%B,/ 0% - 20%A/20%/60% C in 40 minutes.

Example 50

HOOC-CH2-D-Dpa-Pro-Ppa-(2-thiazolyl)

(a) Boc-D-Dpa-Pro-OBzl

Boc-D-Dpa-Pro-OBzl was obtained by reaction of Boc-D-Dpa-OH (4.77 g) with H-Pro-OBzl (3.8 g) in accordance with the reaction conditions of interaction described in Example 1. Output: 8.4V,

TLC: Rf0.95, and silica gel; ethyl acetate/pyridine/acetic acid/water= 560/31/18/7, about/about/about/about.

(b) Boc-D-Dpa-Pro-OH

Boc-D-Dpa-Pro-OH was obtained by the reaction of catalytic hydrogenation of Boc-D-Dpa-Pro-OBzl (7 g) by the method described in Example 1. Output: 5.5V,

TLC: Rf0,59, silica gel; ethyl acetate/pyridine/acetic acid/water= 520/31/18/7, about/about/about/about.

(C) Boc-D-Dpa-Pro-Ppa(Teoc)-(2-thiazolyl)

The reaction of interaction of Boc-D-Dpa-Pro-OH (500 mg) with H-Ppa(Teoc)-(2-thiazolyl). TsOH (573 mg) was performed by the method described in Example 48. Output: 712 mg

TLC: Rf0,44; silica gel, heptane/ethyl acetate=4:6, about/about.

(d) N-(t-butyloxycarbonyl)-D-Dpa-Pro-Ppa(Teoc)-(2-thiazolyl

Boc-D-Dpa-Pro-Ppa(Teoc)-(2-thiazolyl) (712 mg) was treated with para-toluensulfonate acid by the method described in Primem alkylation reaction H-D-Dpa-Pro-Ppa(Teoc)-(2-thiazolyl) (761 mg) with tert-butylbromide (was 0.138 ml) method, described in Example 1. Output: 674 mg

TLC: Rf0,54; silica gel, heptane/ethyl acetate=3:7, V/V.

(e) HOOC-CH2-D-Dpa-Pro-Ppa-(2-thiazolyl)

Subsequent withdrawal of protection and purification was performed by the method described in Example 48, and received two separate diastereoisomer.

Yield: 64 mg Rt: 35,16 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Yield: 96 mg Rt: and 36.80 min, 20%A/80%/0%C to 20%A/20%/60%C in 40 minutes.

Example 51

HOOC-CH2-D-Cha-Ohi-Ppa-(2-thiazolyl)

(a) N-(t-Butyloxycarbonyl)-N-BOC-D-Cha-HE

N-(t-butyloxycarbonyl)-N-Boc-D-Cha-OH was obtained by the method described in Example 45.

(b) H-Ohi-OMe.HCl

To a solution of (-15oC) dry methanol (10 ml) was added dropwise of 0.43 ml (0.7 g) of thionyl chloride. The mixture was stirred for 20 minutes at -10oC, and then added octahedrally-2-carboxylic acid (H-Ohi-OH) (0.5 g) and the solution was heated under reflux for 3 hours. The mixture was concentrated and evaporated with methanol in vacuo, the resulting 665 mg H-Ohi-OMe.HCl

TCX: Rf0,68; silica gel; butanol/pyridine/acetic acid/water=4/1/1/2, about/about/about/about.

(C) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Ohi-OMe

To a cold (0oC) the d) and dicyclohexylcarbodiimide (700 mg) and the mixture was stirred for 20 minutes at 0oC. To this reaction mixture was added H-Ohi-OMe.HCl (665 mg) and N,N-diisopropylethylamine (0.075 ml). The mixture was stirred at 0oC for 1 hour and then kept at room temperature for 7 days, while maintaining a pH=7 by adding N,N-diisopropylethylamine. The mixture was cooled to -20oC and dicyclohexylamine was removed by filtration. The filtrate was evaporated to dryness. The residue was dissolved in ethyl acetate and washed with 1 n hydrochloric acid, saturated sodium bicarbonate and saturated saline solution. Then was dried with sodium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel using as eluent heptane/ethyl acetate=3:1, vol/vol. Output: 1,07 g

TCX: Rf0,41; silica gel, heptane/ethyl acetate=3:1, about/about.

(d) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Ohi-OH

To a solution of N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Ohi-OMe (1,05 g) in dioxane/water 2/1, V/V (30 ml) was added monohydrate of lithium hydroxide (0.32 g). The mixture was stirred at room temperature for 6.5 hours. The dioxane was removed by evaporation under reduced, pressure. And then added ethyl acetate, the pH was brought to 1.5 using hydrochloric acid, and the aqueous layer three times EXT sodium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel using as eluent heptane/ethyl acetate (1:2, V/V). The fractions containing N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Ohi-OH were combined and evaporated. Output: 500 mg.

TLC: Rf0,25; silica gel, heptane/ethyl acetate, 1/2, about/about.

(e) HOOC-CH2-D-Cha-Ohi-Ppa(2-thiazolyl)

The reaction of interaction of N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Ohi-OH and H-Ppa(Teoc)-(2 - thiazolyl).TsOH (428 mg), followed by the removal of protection and purification was performed by the method described in Example 48 and received two separate diastereoisomer.

Yield: 85 mg Rt: 37,72 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Yield: 127 mg Rt: of 42.46 min, 20/A/80%/0% - 20%A/20%/60%C in 40 minutes.

Example 52

HOOC-CH2-D-Cha-Pro(4-CIS-ethyl)-Ppa-(2-thiazolyl)

(a) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro(4-CIS-ethyl)-OE

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-OH was obtained as described in Example 45. To a cold (0oC) a solution of N-(t-butyloxycarbonyl)-N-Boc-D-Cha-OH (3.7 g) in dichloromethane (15 ml) was added 1-hydroxybenzotriazole (1/43 g), dicyclohexylcarbodiimide (2,18 ml) and the mixture was stirred for 15 minutes at 0oC. To this reaction mixture was added H-Pro(4-CIS-ethyl)-OEt. HCl (1,99 g), which was obtained Spa then kept at room temperature for 2 hours, while maintaining a pH=7 by adding N,N-diisopropylethylamine. The mixture was cooled to - 20oC and dicyclohexylamine was removed by filtration. The filtrate was evaporated to dryness. The residue was dissolved in ethyl acetate and washed with 1 n hydrochloric acid, saturated sodium bicarbonate, water and saturated saline, then was dried with sodium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel using heptane/ethyl acetate (3/1, V/V) as eluent. The fractions containing N-(t-butyloxycarbonyl)-N-BOC-D-Cha-Pro(4-CIS-ethyl)-OEt were collected and evaporated. Output a 3.87 g

TLC: Rf0,46; silica gel, heptane/ethyl acetate=3/1, V/V.

(b) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro(4-CIS-ethyl)-HE

To a solution of N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro(4-CIS-ethyl)-OEt (of 3.85 g) in dioxane/water (9/1, V/V) (50 ml) was added 1 n sodium hydroxide (7.5 ml). The mixture was stirred at room temperature. After 60 hours was added 70 mg of the monohydrate of lithium hydroxide in 10 ml of water and this mixture was stirred for another 12 hours. The reaction mixture was diluted with 100 ml water, pH was brought to 1.5 using hydrochloric acid, and the aqueous layer was extracted three times with dichloromethane. The organic layer PR is headed the remainder of was chromatographically on silica gel using as eluent heptane/ethyl acetate (1:2, about/about). The fractions containing N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro(4-CIS-ethyl)-IT was collected and evaporated. Output: 1,39,

TLC: Rf0,79; silica gel; ethyl acetate/pyridine/acetic acid/water= 126/20/11/6, about/about/about/about.

(C) HOOC-CH2-D-Cha-Pro(4-CIS-ethyl)-Ppa-(2-thiazolyl)

The reaction of interaction of N-(t-Butyloxycarbonyl)-N-BOC-D-Cha-Pro(4-CIS-ethyl)-HE and H-Ppa(Teoc)-(2-thiazolyl).TsOH (325 mg), followed by the removal of protection and purification was performed by the method described in Example 48, resulting received two separate diastereoisomer.

Output: 77,5 mg Rt: 36,61 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Yield: 104 mg Rt: 40,05 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Example 53

HOOC-CH2-D-Coa-Pro-Ppa-(2-thiazolyl)

(a) Diethyl-2-acetylamino-2-(cyclooctylmethyl)-malonate

To a cold (0oC) suspension of sodium hydride (are 5.36 g) in dioxane (100 ml) dropwise (slowly) was added to dry ethanol (134 ml) and the mixture was stirred for one and a half hours at 0oC in nitrogen atmosphere. The resulting solution ethylate sodium was added dropwise to a solution of cyclooctatetraene (Example 7 (a)), (27,49 g), sodium iodide (2.0 g) and diethylenetriaminepenta (29,10 g) in dioxane(200 ml)/ethanol(20 ml), and then the reduced pressure, after which was added ethyl acetate, and then washed with water, 5% sodium bicarbonate solution and saturated saline solution, dried by magnesium sulfate and concentrated in vacuum. The residue was chromatographically on silica gel using as eluent heptane/ethyl acetate (7/3, V/V). The fractions containing the desired compound were collected and evaporated. Output: 19,60, TLC: Rf0,53; silica gel, heptane/ethyl acetate= 7/3, about/about.

(b) H-D,L-Coa-OH

Amino acid H-SOA-OH was obtained by heating under reflux solution of diethyl 2-acetylamino-2-(cyclooctylmethyl)-malonate (19,60 g) in 4 n hydrochloric acid/acetic acid (2:1, V/V) (450 ml) for 75 hours. After cooling, the crystals were collected by filtration and washed with diethyl ether. Output: 9,83,

TLC: Rf0,59; silica gel; ethyl acetate/pyridine/acetic acid /water= 63/20/6/11, about/about/about/about.

(C) N-(t-Butyloxycarbonyl)-N-Boc-D,L-Coa-Pro-OBzl

Synthesis of N-(t-butyloxycarbonyl)-N-BOC-SOA-Pro-OBzl was carried out by the methods described in Example 1.

(d) HOOC-CH2-D-Coa-Pro-Ppa-(2-thiazolyl)

N-(t-Butyloxycarbonyl)-N-Boc-Coa-Pro-H was obtained by the reaction of hydrogenation of N-(t-butyloxycarbonyl)-N-Boc-Coa-Pro-OBzl (19,61 g) way, oxycarbonyl)-N-Boc-Coa-Pro-OH was isolated by filtration. The filtrate was evaporated under reduced pressure (yield 9,34 g) and this compound was used for binding to H-Ppa(Teoc)-(2-thiazolyl). TsOH (400 mg) followed by removal of protection and purification as described in Example 48, resulting in two separate diastereoisomer.

Yield: 137 mg Rt: 36,10 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Yield: 139 mg Rt: up 38.94 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Example 54

HOOC-CH2-D-Cha-Azt-Ppa-(2-thiazolyl)

(a) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-OH

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-OH was obtained by the method described in Example 1.

(b) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Azt-OH

Isobutylparaben (0,71 ml) was added dropwise to a cooled (-15oC to -20oC) a solution of N-(t - butyloxycarbonyl)-N-Boc-D-Cha-OH (1,90 g) and N-methylmorpholine (0.6 ml) in 15 ml dichloromethane. After 45 minutes was added H-Azt-OH (0.50 g). After one hour the organic phase was extracted with water and the organic layer was dried with sodium sulfate, filtered and the solvent was removed under pressure. Yield: 2.3 g of target compound in the form of almost colorless oil.

FAB-MS: m/e=469 (M+H+)

(C) HOOC-CH2-D-Cha-Azt-Ppa-(2-thiazolyl)

The binding of H-Ppa(Teoc)-(2-enous is but single diastereoisomer.

Yield: 72 mg Rt: 28,10 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Yield: 88 mg Rt: 31,46 min, 20%A/80%/0% - 20%A/20%/60%C in 40 minutes.

Example 55

HOOC-(CH2)2-D-Cha-Pro-Ppa-(2-thiazolyl)

(a) N-(2-(t-Butyloxycarbonyl)ethyl)-D-Cha-OMe

To a suspension 7,11 g D-Cha-OMe.HCl in 25 ml of acetonitrile was added to 2.6 ml of N, N-diisopropylethylamine and 25 ml of tert-butyl acrylate. This reaction mixture was heated at 40oC and maintained at pH=8 (test spot held on damp indicator paper to determine the pH) using N,N-diisopropylethylamine. After one week, the reaction mixture was concentrated. The residue was dissolved in ethyl acetate, washed with water and saturated saline, then was dried (sodium sulfate) and concentrated to obtain 11,78 g of target compound, which was used without further purification.

TLC: Rf0,7, silica gel, ethyl acetate/heptane=1:1, about/about.

(b) N-(2-(t-Butyloxycarbonyl)ethyl)-N-Boc-D-Cha-OMe

The method described for N-(t-butyloxycarbonyl)-N-Boc-D-Cha-OMe (Example 1), N-(2-(t-butyloxycarbonyl) ethyl)-D-Cha-OMe (11,78 g) was converted into N-(2-t-butyloxycarbonyl) ethyl)-N-Boc-D-Cha-OMe (8,78 g)

TLC: Rf0,7, silica gel, ethyl acetate/heptane=1:2, about/about.


TLC: Rf0,1, silica gel, ethyl acetate/heptane=1/5, about/about.

(d) N-(2-(t-Butyloxycarbonyl)ethyl)-N-Boc-D-Cha-Pro-OBzl

In a manner analogous to that described for N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro-OBzl (Example 1), N-(2-(t-butyloxycarbonyl)ethyl)-N-Boc-D-Cha-OH (3,14 g) was subjected to interaction with Pro-OBzl.HCl, resulting in a received N-(2-(t-butyloxycarbonyl)ethyl)-N-Boc-D-Cha-Pro-OBzl (4,36 g).

TLC: Rfof 0.5, silica gel, ethyl acetate/heptane=1/2, about/about.

(e) N-(2-(t-Butyloxycarbonyl)ethyl)-N-Boc-D-Cha-Pro-OH

In a manner analogous to that described for N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro-OH (Example 1), N-(2-(t-butyloxycarbonyl)ethyl)-N-Boc-D-Cha-Pro-OBzl (4,33 g) was subjected to hydrogenation reactions, resulting in a received N- (2-(t-butyloxycarbonyl)ethyl)-N-Boc-D-Cha-Pro-OH (3,96 g)

TLC: Rf0,15, silica gel, ethyl acetate.

(f) N-(2-(t-Butyloxycarbonyl)ethyl)-N-Boc-D-Cha-Pro-PPA(Teoc) -(2)-thiazolyl)

In a manner analogous to that described for N-(t-butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Teoc)-(2-oxazolyl) (Example 44), N-(2-(t-butyloxycarbonyl) ethyl)-N-Boc-D-Cha-Pro-OH (342 mg) was subjected to a reaction between H - Ppa(Teoc)-(2-thiazolyl). TsOH (description of the LASS="ptx2">

TLC: Rf0.25, silica gel, ethyl acetate/heptane=1/1, V/V.

(g) HOOC-CH2CH2-D-Cha-Pro-Ppa-(2-thiazolyl)

In a manner analogous to that described for HOOC-CH2-D-Cha-Pro-PPA-(2-oxazolyl) (Example 44), N-(2-(t-butyloxycarbonyl) ethyl)-N-Boc-D-Cha-Pro-Ppa (Teoc)-(2-thiazolyl) (468 mg) was subjected to reactions release and received N-NOO-CH2CH2-D-Cha-Pro-Ppa-(2-thiazolyl) in the form of two diastereoisomers.

Yield: 123 mg Rt(LC): 30,2 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Yield: 138 mg Rt (LC): 33,6 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 56

HOOC-CH2-(N-Me)-D-Cha-Pro-Ppa-(2-thiazolyl)

(a) Boc-D-Cha-OMe

Boc-D-Cha-OH (20 g) was dissolved in dichloromethane/methanol (9/1, V/V) (400 ml). Then add tetrafluoroborate 2-(1H - benzotriazol-1-yl)-1,1,3,3-tetramethylurea (22,7 g) and the solution was brought to pH=8 by addition of triethylamine (10 ml). The reaction mixture was stirred at room temperature for 16 hours. The mixture is then washed with cold 1 n hydrochloric acid, water, 5% sodium bicarbonate and water, then dried with sodium sulfate. The filtrate was evaporated and the residue was chromatographically on silica gel using as eluent ethyl acetate/heptane = 2/3, about/about. Faction, with whom, 3/1, about/about.

(b) Boc-(N-Me)-D-Cha-OMe

Boc-D-Cha-OMe (19,35 g) was dissolved in 200 ml of dry N,N-dimethylformamide in a nitrogen atmosphere. After you have added under the conditions (4,22 ml) and the mixture was cooled to 0oC. Then was added a 2.71 g of sodium hydride (60% dispersion in oil) and the reaction mixture was stirred for 3 hours at room temperature. This mixture was partially concentrated, added ethyl acetate and the organic layer was washed with 0.1 M hydrochloric acid, water, 5% sodium bicarbonate solution and saturated saline, then was dried with sodium sulfate and concentrated. Output: 21,07,

TLC: Rfof 0.55, silica gel, ethyl acetate/heptane, 3/7, about/about.

(C) Boc-(N-Me)-D-Cha-OH

A solution of Boc-(N-Me)-D-Cha-OMe (20 g) in 400 ml of dioxane/water (9/1, V/V) was treated with a sufficient amount of a 2 M solution of sodium hydroxide to maintain pH= 12 for 16 hours at room temperature. After acidification, the mixture was poured into water and was extracted with dichloromethane. The organic layer was washed with water and dried with sodium sulfate. The filtrate was evaporated and got to 21.2 g of Boc-(N-Me)-D-Cha-OH.

TLC: Rf of 0.75; silica gel; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(d) Boc-(N-Me)-D-Cha-OBzl

Boc-(N-Me)-D-Cha-OH (2 g) was dissolved in tilorone (2.24 g), then the solution was brought to pH=8 by addition of triethylamine (10 ml). The reaction mixture was stirred for one hour at room temperature. The mixture is then washed with cold 1 n hydrochloric acid, water, 5% sodium bicarbonate and again with water, then dried with sodium sulfate. The filtrate was evaporated and the residue was chromatographically on silica gel using ethyl acetate/heptane (1: 3, V/V) as eluent. The fractions containing (N-Me)-Boc-D-Cha-OBzl was collected and evaporated. Output: 1,56,

TLC: Rf0.75, silica gel, ethyl acetate/heptane=3/1, V/V.

(e) H-(N-Me)-D-Cha-OBzl

Boc-(N-Me)-D-Cha-OBzl (1.56 g) was treated with 50% triperoxonane acid/dichloromethane (20 ml) for 1 hour at room temperature. The reaction mixture was evaporated to dryness and received 1.6 g of the target compound.

TLC: Rfof 0.50, silica gel; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(f) N-(t-Butyloxycarbonyl)-(N-Me)-D-Cha-OBzl

To a stirred solution of H-(N-Me)-D-Cha-OBzl (1.6 g) in 40 ml of acetonitrile was added t-butylbromide (0,81 g). Then add tetrabutylammonium iodide (150 mg) and the pH of the mixture was brought to 8.5 by adding N,N-diisopropylethylamine. The resulting mixture peremeci is whether 5% sodium bicarbonate solution and water, and then was dried with sodium sulfate and evaporated in vacuum. After chromatography on silica gel (eluent: heptane/ethyl acetate= 4/1, V/V) was received of 1.11 g of N-(t-butyloxycarbonyl)-(N-Me)-D-Cha-OBzl.

TLC: Rf0.95, and silica gel; ethyl acetate/pyridine/acetic acid/water = 63/20/6/11, about/about/about/about.

(g) N-(t-Butyloxycarbonyl)-(N-Me)-D-Cha-OH

To a solution of N-(t-butyloxycarbonyl)-(N-Me)-D-Cha-OBzl (1,11 g) in methanol (20 ml) was added 10% palladium-on-coal (100 mg) and 1.43 ml of 2 M hydrochloric acid. The reaction mixture was first made at atmospheric pressure and at room temperature for 1 hour. The palladium catalyst was removed by filtration and the solvent was removed by evaporation under reduced pressure to obtain 970 mg of N-(t-butyloxycarbonyl)-(N-Me)-D-Cha-OH.

TLC: Rf0.75, silica gel; ethyl acetate/pyridine/acetic acid/water= 163/20/6/11, about/about/about/about.

(h) HOOC-CH2-(N-Me)-D-Cha-Pro-Ppa-(2-thiazolyl)

N-(t-Butyloxycarbonyl)-(N-Me)-D-Cha-Pro-OH was obtained by the method described in Example 1. Linking using a mixed anhydride, removing the protection and purification was performed by the method described in Example 48. Output: two diastereoisomer, both 79 mg.

A: Rt(LC): 30,87 min, 20%A/80% - 20%A/20%/p 57

EtOOC-CH2-D-Cha-Pro-Ppa-(2-thiazolyl)

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Pro-Ppa(Teoc)-(2-thiazolyl) was obtained by the method described in Example 48. Removing protection was performed using 3 M hydrogen chloride/ethanol. After cleaning, received two diastereoisomer: 37 mg and 44 mg

A: Rt(LC): 35,24 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

IN: Rt(LC): 38,12 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 58

H-(N-Me)-D-Cha-Pro-Ppa-(2-thiazolyl)

Boc-(N-Me)-D-Cha-Pro-OH

Synthesis of Boc-(N-Me)-D-Cha-Pro-OH as described in Example 16.

N-Me-D-Cha-Pro-Ppa-(2-thiazolyl)

The methods described in Example 45, Boc-(N-Me)-D-Cha-Pro-OH (400 mg) was subjected to binding with H-Ppa(Cbz)-(2-thiazolyl).TFA Derived Tripeptide was treated in a mixture of TFA and thioanisole (see Example 45) and was purified by HPLC to obtain the target compound (218 mg) as a mixture of diastereoisomers.

Rt(LC): 31,96 min, 20%A/80% - 20%A/20%/60%C in 40 minutes.

Example 59

SO2-D-Sha-Pro-PPA(2-thiazolyl)

(a) SO2>-D-Cha-Pro-HE

Synthesis SO2-D-ha-Pro-HE described in Example 18.

b) SO2-D-Cha-Pro-Ppa-(2-thiazolyl)

Linking SO2-D-Cha-Pro-OH (300 mg) with H-Ppa(Cbz)-(2-thiazolyl) were carried out by the method described in Example 45, the floor is of Example 45, getting SO2-D-Cha-Pro-Ppa-(2-thiazolyl) (190 MT) as a mixture of diastereoisomers.

Rt(LC): 26,67 and 28,20 min, 20%A/60%b and 20%C - 100%C for 40 minutes.

Example 60

SO2-D-Phe-Pro-Ppa-(2-thiazolyl)

Linking SO2-D-Phe-Pro-OH (Example 26) with H - Ppa(Teoc)-(2-thiazolyl). TsOH (Example 47) was carried out by the methods described in example 48, and got fully protected Tripeptide. After removal protection (see Example 48), two diastereoisomer can be separated by using HPLC. Output: two diastereoisomer.

80 mg: Rt(LC): 39,87 min, 20%A, 80% - 20%A, 20%b and 60%C in 40 minutes.

69 mg: Rt(LC): ø 42.45 min, 20%A, 80% - 20%A, 20%b and 60%C in 40 minutes.

Example 61

SO2-noeLeu(cyclo)-Gly-Ppa-(2-thiazolyl)

235 mg SO2-noeLeu(cyclo)-Gly-OH (see Example 12) were subjected to binding with H-Ppa(Cbz)-(2-thiazolyl).TFA under the same conditions as described in Example 45. After removal of the protective group from the resulting Tripeptide (example 45) and after HPLC purification were obtained from two separate diastereoisomer.

Yield: 99 mg Rtt (LC): 37,99 min, 20%A, 80% - 20%A, 20%b and 60%C in 40 minutes.

Yield: 86 mg Rt(LC): 38,68 min, 20%A, 80% - 20%A, 20%b and 60%C in 40 minutes.

Example 62

Firmly the ru 18,4 g H-Ppa-OH and 6.8 g of the pentahydrate of copper sulfate (2) in water (312 ml) and dioxane (234 ml) was added to 33.9 g of di-t-BUTYLCARBAMATE, leading up to pH=9 by addition of 4 n sodium hydroxide. The mixture was stirred for 16 hours at room temperature. The precipitate was collected and washed with water, sediment on the filter is suspended in dioxane and the pH brought to 12.5 by addition of 4 n sodium hydroxide. Then was added 24 ml of 2-trimethylsilylamodimethicone two portions with an interval of 0.5 hours. The mixture was stirred for 16 hours. The precipitate was filtered and washed with dioxane. The filtrate was concentrated to small volume and the pH was brought to 2.5. The mixture was diluted with ethyl acetate, washed with water, dried with sodium sulfate and was evaporated to dryness to obtain 24,9 g butter.

TLC: Rf0,79, silica gel; ethyl acetate/pyridine/acetic acid/water= 63/20/6/11, about/about/about/about.

(b) Teoc-Ppa(Boc)-N(Me)OMe

To a solution 29,8 g Teoc-Ppa(Boc)HE and rate of 7.54 g of the hydrochloride of N,O-dimethylhydroxylamine in 370 ml of dichloromethane was added 23 g of tetrafluoroborate O-(benzotriazol-1-yl)-N,N,N',N'-tetramethylurea and 20.2 ml of triethylamine and the mixture was stirred for 2 hours. The reaction mixture was extracted with water and the aqueous 0.9 M sodium bicarbonate, and then dried with magnesium sulfate and concentrated. The residue was chromatographically on silica gel using 0--->5% methanol in dichloromethane, real.

(C) Teoc-Ppa(Boc)-(2-thiazolyl)

70 ml of n-Utility (0,72 M in diethyl ether) was added dropwise to a cooled (-78oC) the solution 4,55 ml just subjected to distillation 2-bromothiazole in 40 ml of diethyl ether. The reaction mixture was stirred for another 15 minutes, and then slowly added 7.2 g Teoc-Ppa(Boc)-N(Me)OMe dissolved in 40 ml of tetrahydrofuran. After stirring for 30 minutes the reaction mixture was poured into aqueous sodium bicarbonate solution (200 ml). The ether layer was separated and the aqueous was extracted with ether. The combined ether extracts were washed with water, dried with magnesium sulfate and concentrated. The residue was purified by column chromatography on silica gel and got to 6.43 g of the target compound.

TLC: Rf0,61; silica gel, toluene/acetone=3/1, V/V.

(C) Teoc-Ppa-(2-thiazolyl).TsOH

3,16 g p-toluensulfonate acid (1 H2O) was added to a solution of 8 g of Teoc-Ppa(Boc)-(2-thiazolyl) in 116 ml of diethyl ether. The resulting mixture was slowly concentrated on a rotary evaporator at 30oC. the Oily residue was stirred another 2 hours on a rotary evaporator under vacuum, resulting in 9.2 grams of salt p-toluensulfonate acid.

TLC: Rf0.6, silica gel; ethyl acetate/petrolatina (Teoc-PPP(C(O)OCH2Po1)-(2-thiazolyl)

To a suspension of 10.0 g of hydroxymatairesinol (HOCH2Ro) -(0,97 mm/g) and 12.9 g of di(N-Succinimidyl)carbonate in acetonitrile (50 ml) and dichloromethane (50 ml) was added 7 ml of triethylamine. The resulting mixture was shaken for 2 hours at room temperature. The resin was filtered and then washed with dichloromethane, 1-methyl-2-pyrrolidinone and again dichloromethane. This resin was suspended in acetonitrile (50 ml) and dichloromethane (50 ml). Then, to the suspension was added to 8.3 g Teoc-Ppa-(2-thiazolyl).TsOH and 3.7 ml of triethylamine and the resulting mixture was shaken for 17 hours. The resin was collected by filtration, then washed with dichloromethane, 1-methyl-2-pyrrolidinone, dichloromethane and diethyl ether, then dried under vacuum and obtained 13.3 g of resin.

(f) H-Ppa(C(O)OCH2Pol)-(2-thiazolyl).HCl

1.5 ml of the mixture triperoxonane acid and dichloromethane (1:1, V/V) was added to 50 mg Teoc-PPP(C(O)OCH2Pol)-(2-thiazolyl) and the reaction mixture was shaken for 30 minutes. The resin was filtered, and then washed with dichloromethane, 0.5 M Hcl in dioxane-dichloromethane (1:7, V/V) and dichloromethane.

(g) Boc-Phe-PPP(C(O)OCH2Pol)-(2-thiazolyl)

To a solution of 40 mg of Boc-Phe-OH and 20 mg of 1-hydroxybenzotriazole, 1-methyl-2-pyrrolidone (1 the Olu. Then add 5 ál of the 4-methylmorpholine and the resulting suspension was shaken for 2 hours, filtered, and then washed with 1-methyl-2-pyrrolidinone, dichloromethane and diethyl ether. Other N-t-butyloxycarbonyl acid were added in the same way.

(h) H-Phe-PPP(C(O)OCH2Po1)-(2-thiazolyl).HCl

Removal of N-t-butyloxycarbonyl group from Boc-Phe-PPP(C(O)OCH2Po1)-(2-thiazolyl) was carried out in a manner analogous to that described above for removal of the Teoc group at Teoc-PPP(C(O)OCH2Po1)-(2-thiazolyl)

(i) N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-Phe-PPA(C(O)OCH2Pol)-(2-thiazolyl)

N-(t-Butyloxycarbonyl)-N-Boc-D-Cha-OH was attached to H-Phe-PPP(C(O)OCH2Po1)-(2-thiazolyl). HCl manner similar to that described above for attachment of Boc-Phe-OH to H-Ppa(C(O)OCH2Po1)-(2-thiazolyl).

(j) HOOC-CH2-D-Cha-Phe-Ppa-(2-thiazolyl)

The resin was treated with 1 ml of a mixture of triperoxonane acid/thioanisole (10/1, V/V) and shaken for 4 hours. The reaction mixture was filtered, and then washed triperoxonane acid and toluene. The combined filtrate and washings were concentrated. To the residue was added heptane and the mixture was subjected to intense vortex mixing. Heptane layer was removed, and was nerastvorimoe through a filter of 0.45 μm and liofilizovane. The crude peptide was purified using preparative pressure chromatography (column: Superdex peptide HR 10/30; flow rate: 0.75 ml/min; eluent: water/acetonitrile= 4/1, V/V). After lyophilization of the appropriate fractions were obtained 6.4 mg HOOC-CH2-D-Cha-Phe-Ppa-(2-thiazolyl)

(k) HOOC-CH2-D-Cha-X-Ppa-(2-thiazolyl) (table 62)

The compounds of formula HOOC-CH2-D-Cha-X-Ppa-(2-thiazolyl) was obtained by methods similar to get HOOC-CH2-D-Cha-Phe-PPA-(2-thiazolyl). All compounds were characterized using reverse-phase liquid chromatography on a column with LC-18-DB Spelcosil under the following conditions: flow rate: 1.0 ml/min, buffer A: 0.5 M phosphate potassium buffer (pH 2,1); buffer A: water; buffer D: acetonitrile-water (9:1). Gradient: 0 ---> 30 min: 20% A/65%/15% D ---> 20%/25%/55% D. UV-detection at 210 nm. Table 62 retention time is given in minutes.

Example 63

Using the methods of the present invention can be obtained the following compounds

N-Me-D-Phe-Pro-Ppa-(2-thiazolyl)

HOOC-CH2-D-Phe-Pro-Ppa[COCO]-

HOOC-CH2-D-Phe-Pro-Ppa-(2-thiazolyl)

HOOC-CH2-D-Cha-(N-cyclopentyl)-Gly-Ppa-(2-thiazolyl)

HOOC-CH2-D-Cha-Pec-Ppa[COCO]-

HOOC-CH2-D-Cha-Pec-Ppa-(2-thiazolyl)

HOOC-CH2-D-Cha-(N-cyclohexyl)-Gly- HOOC-CH2-D-Cha-(N-cyclopropyl)-Gly-Ppa-(2-thiazolyl)

N-Me-D-Phe-(N-cyclopentyl)-Gly-Ppa[COCO]-OH

N-Me-D-Phe-(N-cyclopentyl)-Gly-Ppa-(2-thiazolyl)

2-Propyl-pentanoyl-Asp(OMe)-Pro-PPA[COCO]-

2-Propyl-pentanoyl-Asp(OMe)-Pro-Ppa-(2-thiazolyl)

2-Propyl-pentanoyl-Asp-Pro-Ppa[COCO]-OH

2-Propyl-pentanoyl-Asp-Pro-Ppa-(2-thiazolyl)

1-Piq-(N-cyclopentyl)-Gly-Ppa[COCO]-OH

1-Piq-(N-cyclopentyl)-Gly-Ppa-(2-thiazolyl)

Diphenylpropionic-Pro-Ppa-(2-thiazolyl)

N-Me-D-Nle-Pro-Ppa[COCO]-OH

N-Me-D-Nle-Pro-Ppa-(2-thiazolyl)

EtSO2-D-Phe-Pro-Ppa[COCO]-OH

EtSO2-N(Me)-D-Cha-Pro-Ppa[COCO]-OH

EtSO2-N(Me)-D-Cha-Pro-Ppa-(2-thiazolyl)

EtSO2-N(Me)-D-Cha-Pro-Ppa-(2-oxazolyl)

HOOC-CH2-N(Me)-D-Cha-Pro-Ppa[COCO]-OH

HOOC-CH2-N(Me)-D-Cha-Pro-Ppa-(2-thiazolyl)

HOOC-CH2-N(Me)-D-Cha-Pro-Ppa-(2-oxazolyl).

Example 64

Analysis on antithrombotic activity

Thrombin (Factor IIA) is a factor involved in the cascade of reactions of blood coagulation. Antitromboticescoy activity of the compounds of the present invention was assessed by spectrophotometric measurement of the rate of hydrolysis of a chromogenic substrate s-2238, manifested by the action of thrombin. This analysis on antithrombotic activity held in the buffer system used to estimate the IC5soy is likely 6000 (TNP). Known connection: 12581 (Kabi). Media: TNP buffer. Solubilization can be carried out using dimethyl sulfoxide, methanol, ethanol, acetonitrile or tert-butyl alcohol, which no adverse effects at concentrations up to 2.5% in the final reaction mixture.

Technique

Reagents*: Tromethamine-NaCl (TN) buffer. The composition of this buffer: tromethamine (Tris) - 6,057 (50 mmol), NaCI - 5,844 g (100 mmol), water to 1 l, pH of the solution was brought to 7.4 at 37oC by adding HCl (10 mmol/l). TNP buffer: polyethylene glycol 6000 was dissolved in TN buffer with obtaining a concentration of 3 g/l Solution of S-2238: one vessel S-2238 (25 mg; Kabi Diagnostica, Sweden) was dissolved in 20 ml of TN buffer with obtaining a concentration of 1.25 mg/ml (2 mmol/l). A solution of thrombin: human thrombin (16000 ncat/vessel; Central Laboratorium voor Bloedtransfusie, Amsterdam, The Netherlands) was dissolved in TNP buffer and received the mother solution with a concentration of 835 ncat/ml Immediately before use, this solution was diluted with TNP-buffer with obtaining concentration 3,34 ncat/ml.

-*All ingredients used were of analytical purity.

For aqueous solutions used ultrapure water (Milli-Q).

To obtain solutions of the subjects of the i-Q to obtain a concentration of 10-2mol/l of Each concentration was serially diluted with a carrier to obtain concentrations of 10-3, 10-4and 10-5mol/L. These cultivation, including the mother liquor used in the analysis (final concentration of the reaction mixture was 310-3; 10-3; 310-4; 10-4; 310-5; 10-5; 310-6; and 10-6mol/l, respectively).

Procedure

At room temperature 0.075 ml and 0.025 ml of the test solutions or known compounds or media alternative was introduced by pipette into the wells of the microplate, and these solutions were diluted 0,115 ml and 0,0165 ml TNP-buffer, respectively. Then to each well was added 0,030 ml-aliquot of the solution of S-2238 and the plate was pre-heated and pre-incubated with shaking in an incubator (Amersham) for 10 minutes at 37oC. After pre-incubation was initiated by the hydrolysis of S-2238 by adding to each well 0,030 ml thrombin. The tablet then incubated (with shaking for 30 s) at 37oC. After 1 minute after the start of incubation every 2 minutes for 90 minutes was measured optical density of each sample at 405 nm using a kinetic microplate reader (Twinreader plus, Flow Laborator the radio connection (expressed in mol/l of the reaction mixture) and for each "control" built curve of the dependence of optical density on reaction time (min.)

Assessment results: Based on the analytical curve for each final concentration was calculated maximum optical density. The value of the IC50(the final concentration in µmol/l causing 50% inhibition of the maximum optical density of control) was calculated using the analysis method of the logarithmic transformation of Hafner (Hafner et al., Arzneim.-Forsch/Drug Res. 1977; 27(11): 1871-3).

The value of the IC50for compounds of the present invention are listed in the following table A.

1. The compound of the formula I

< / BR>
where A is H, optionally substituted D, L--hydroxyacetic, R1, R1-O-C(O)-, R1-C(O)-, R1SO2-, R2OOC-(CHR2)m- where R1selected from (1-12C)alkyl and (3-8C)cycloalkyl, and these groups may be substituted by an OH group, and from (7-15C)aralkyl, and each group R2independently represents H or has the same values that are specified for R1; m = 1, 2, or 3;

B represents a bond, an amino acid of formula-NH-CH[(CH2)pC(O)OH]-C(O)-, where p = 0, 1, 2, or 3, -N(( 1-12C)alkyl)-CH2-CO-D-1-Piq or B is a D-amino acid having a hydrophobic side chain, where the specified hydrophobic side chain is (1-12C)alkyl, optionally exil, cyclooctyl, phenyl, and where specified hydrophobic side chain may be optionally substituted by such substituents as halogen, lower alkyl (e.g. methyl or ethyl), lower alkoxy (e.g. methoxy), or B represents L - or D-amino acid having a basic side chain, and the specified hydrophobic or basic amino acid may be optionally substituted N-(1-6C)alkyl;

either A and B taken together represent the residue R3R4N-CHR5-C(O)-, where R3and R4independently represent R1, R1-O-C(O)-, R1-C(O)-, R1-SO2-, and R5is a hydrophobic or basic side chain;

X represents L-amino acid with a hydrophobic side chain, where this side chain is (1-12C)alkyl, optionally substituted by one or more (6-14C)aryl groups (e.g. phenyl), or X represents a threonine, or X represents a cyclic amino acid, such as 2-azetidinone acid, Proline, 4-piperidinylcarbonyl acid, 2-octahydrocyclopenta acid where the specified cyclic amino acid optionally substituted (1-6C)alkyl, or X is-NR2-CH2-C(O) -- or the fragment

< / BR>
or

< / BR>
where n = 2, 3, or 4, and W , CONR7R8where R7and R8independently - H or (1-6C)alkyl, or R7and R8taken together, represent (3-6C)alkylene, or Y is a heterocycle selected from 2-thiazole, 2-benzothiazole, 2-oxazole or 2-benzoxazole;

r = 0, 1, 2 or 3,

or its pharmaceutically acceptable salt.

2. Connection on p. 1, where X Is an L-amino acid with a hydrophobic side chain, threonine or-NR2-CH2-C(O)-.

3. Connection on p. 1, where A is defined above; B is a bond, an amino acid of formula-NH-CH[(CH2)pC(O)OH]-C(O)-, where p = 0, 1, 2 or 3; -N((1-6C)alkyl)-CH2-CO-D-1-Piq or a D-amino acid having a hydrophobic side chain, where the specified amino acid may be optionally substituted N-(1-6C)alkyl, or A and B taken together represent R3R4N-CHR5-C(O)-, and X represents a cyclic amino acid, optionally substituted (1-6C)alkyl, or X is-NR2-CH2-C(O) -- or the fragment

< / BR>
or

< / BR>
4. Connection on p. 3, where A is H, 2-hydroxy-3-cyclohexylpropionic, R1, R1-SO2-, R2OOC-(CHR2)m- where R1selected from (1-12C)alkyl and (7-15C)aralkyl; B - link, D-l-Piq or a D-amino acid having a hydrophobic side chain, where the specified amino acid R5-C(O)-.

5. Connection on p. 4, where A is H, R1-SO2or R2OOC-(CHR2)m-; B is a bond, D-1-Piq or a D-amino acid having a hydrophobic side chain, or A and B taken together represent R3R4N-CHR5-C(O)-; Y IS-CO-NH-(1-6C)alkylene-C6H5, -COOR6where R6Is H or (1-3C)alkyl, -CONR7R8where R7and R8independently represent H or (1-3C)alkyl, or R7and R8taken together, represent a (3-5C)alkylene, or Y is a heterocycle selected from 2-thiazole, 2-benzothiazole, 2-oxazole or 2-benzoxazole.

6. Connection on p. 5, where A is R2OOC-(CHR2)m-; B - D-amino acid having a hydrophobic side chain, or A and B taken together is the remainder of R3R4N-CHR5-C(O)-, and X - 2-azetidinone acid, Proline, 2-octahydrocyclopenta acid or [N(3-8C)cycloalkyl]-CH2-C(O)-.

7. Connection on p. 6, where A IS HOOC-CH2-; B - D-Phe, D-Cha, D-Coa, D-Dpa, p-Cl-D-Phe, p-o-methyl D-Phe, p-Outil-D-Phe, D-Nle, m-Cl-D-Phe, 3,4-di-OMe-D-Phe, D-Chq or A and B taken together, R3R4N-CHR5-C(O)-;

8. Connection on p. 7, where A IS HOOC-CH2-; B - D-Cha; X is Proline or -[N(cyclopentyl)]-CH2-C(O)-.

9. Connection on p. 5, where A is R1-SO2-; B - link, D-l-Piq or a D-aminona least one of R3and R4- R1-SO2- and the other independently is (1-12C)alkyl, or R1-SO2- X - 2-azetidinone acid, Proline, 2-octahydrocyclopenta acid, -[N(3-8C)cycloalkyl]-CH2-C(O) -- or the fragment

< / BR>
or

< / BR>
10. Connection on p. 9, where A - ethyl-SO2- or benzyl-SO2-; B - link D-Phe, D-Cha, D-Coa, D-Dpa, p-Cl-D-Phe, p-o-methyl D-Phe, p-Outil-D-Phe, D-Nle, m-Cl-D-Phe, 3,4-di-OMe-D-Phe, D-Chq or A and B taken together, the balance3R4N-CHR5-C(O)-, where at least one of R3and R4- ethyl-SO2or benzyl - SO2- and the other independently is (1-12C)alkyl, or R1-SO2and R5is (3-8C)cycloalkyl, (3-8C)cycloalkyl(1-4C)alkyl, phenyl, benzyl, diphenylmethyl, where these groups may be optionally substituted by chlorine or (1-4C)alkoxy.

11. Connection on p. 10, A - ethyl-SO2-, B - D-Cha; X is Proline or[(N(cyclopentyl)]CH2-C(O)-.

12. The compound according to any one of paragraphs.1 - 11, where r = 1.

13. The compound according to any one of paragraphs.1 - 12 used in therapy.

14. The compound according to any one of paragraphs.1 - 12 for the manufacture of a medicinal product for the treatment or prevention of diseases mediated by the action of thrombin.

15. Pharmaceutical composition having antithrombin

 

Same patents:

The invention relates to biotinyl compounds of General formula

< / BR>
where Q is absent or represents-NH-(CH2)5-CO-,

R1denotes X-Arg-Gly-Asp-y,

X - Tripeptide: Gly-Gly-Gly-,

y - dipeptide: -Ser-PrO

or R1denotes A-Cys(R2)-B-U,

R2denotes H, Trt;

And represents Asp or peptide fragment selected from the group consisting of:

Ala-Asp, Thr-Ala-Asp, Lys-Thr-Ala-Asp, Lys-Ala-Ala-Asp,

Arg-Thr-Ala-Asp, Ser-Ala-Asp, Gln-Ser-Ala-Asp,

Gly-Lys-Thr-Ala-Asp, Ile-Ser-Ala-Gly, Arg-Ser-Ala-Gly

Gly-Lys-Thr-Cys (Trt)-Asp

In tstststs or represents Pro or N-methylated derivative of Ala,

moreover, if R1denotes A-Cys(R2)-B-U, only one of the residues a or b may not appear,

U indicates HE or NH2,

or R1means cyclo (Arg-Gly-Asp-Z), Z in the side chain is linked to Q, or if Q is absent, with Biotin,

Z denotes a di - or tripeptides the rest,

moreover, amino acids, independently of one another, are selected from the group consisting of Ala, Val, Tight, Trp, Phe, Cys, Lys, M, these amino acids can be derivatization, and amino acid residues are linked to each other peptidome through N-amino - and = R6- R4< / BR>
where R4= HE

R6= alkylether with 7-14 C atoms,

R8= H, alkyl with 1-6 C-atoms,

moreover, if we are talking about the balance of optically active amino acids and amino acid derivatives, are included as D-, L-forms;

and their salts

The invention relates to bicyclic compounds useful as drugs, the neutralizing effect of glycoprotein IIb/IIIa, to prevent thrombosis

The invention relates to medicine and can be used to adjust parameters of the system of hemostasis

The invention relates to a drug intended for intravenous

The invention relates to new cyclic amine derivatives of General formula I, where R1represents a phenyl group substituted by halogen atom,2represents C1- C8aliphatic acyl group or (C1- C4alkoxy)carbonyl group, R3represents a 3 - to 7-membered saturated cyclic amino group which may form a condensed ring, where the specified cyclic amino group substituted by the Deputy selected from the group comprising: mercaptopropyl, which can be unprotected or protected by a group selected from a number of protective groups, C1- C4alkyl group, substituted mercaptopropyl, which can be unprotected or protected by a group selected from a number of protective groups, and the number of protective groups for the specified mercaptopropyl includes C1- C20alcoholnye group, C3- C20alkenone group and benzoline group, and the said cyclic amino group, furthermore preferably a substituted group of the formula =CR4R5where R4represents a hydrogen atom, and R5represents a hydrogen atom, a C1- C4alkyl group, carboxypropyl, (C1- C4-alkoxy)carbonyl GRU

The invention relates to medicine

The invention relates to medicine, namely to drugs and compositions for the treatment of surgical wounds, burns, lesions, erosions, trophic ulcers, bedsores, etc., especially complicated by purulent-inflammatory processes

The invention relates to medicine, in particular to pharmacology, and in particular to methods of obtaining drugs based on peptides, normalizing the function of various organs and tissues

The invention relates to acceleration of the reaction of nitric oxide in heterogeneous environments, including

The invention relates to medicine and relates to a method of evaluation of snake venom on the presence or absence of platelet aggregation (PA1), based on the specific binding with receptor purified and isolated from snake venom PA1 and its shortened standards, allocation method PA1 from snake venom, and pharmaceutical compositions based on it
The invention relates to the use of CGRP antagonist in a cosmetic, pharmaceutical or dermatological compositions, especially for topical application, for the treatment (processing) herpes, poonsuk, pruriginous of taxidermy and heavy Zudov, as well as new, these compositions
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