NL8202472A - PROLINE DERIVATIVES AND METHOD FOR THE PREPARATION THEREOF. - Google Patents

Description

V. * * V * i I16 -1- 225 ^ 7 / Vk / mb

Short designation: Proline derivatives and method for their preparation.

The invention relates to proline derivatives. The invention further relates to a process for the preparation of proline derivatives.

The proline derivatives according to the invention are characterized in that they correspond to formula 1 stated on the formula sheet, wherein R is an acyl group attached to the oC. amino group of a 10 amino acid and selected from the group consisting of cyclopropanecarbonyl, cyclohexanecarbonyl and adamantane carbonyl groups, A is glycine, sarcosine or oC-D-amino acid residue and with a carbonyl group forming a thiol ester bond with a sulfur atom, and pharmaceutically acceptable salts thereof.

Compounds similar to these compounds represented by formula 1, for example compounds where R is a benzoyl, acetyl or t-butyloxycarbonyl group and A is L-o-amino acid have been described in British patent application 2,050,359A. European patent application 9,898A1 describes compounds wherein R is a benzoyl, acetyl, t-butyloxicarbonyl, cyclopentanecarbonyl group or the like and A is an L-oC amino acid. European patent application 35,383A1 describes compounds in which R is a benzoyl group and A is a D-phenyl-alanyl group. However, there is no publication specifically describing the compounds represented by formula 1 listed on the formula sheet. Therefore, these compounds can be considered as new compounds and are suitable as pharmaceuticals as will be explained in more detail below.

The amino acid residue represented by A in formula 1 is glycine, sarcosine or oG-D amino acid residue. Examples of oC-D amino acids include neutral, acidic or basic amino acids or aliphatic, aromatic, heterocyclic or alicyclic amino acids such as D-alanine, D-leucine, D-asparagine, D-methionine, D-glutamine, D-phenylalanine , D-tryptophan, D-ornitine, D-phenylglycine, D-threonine, D-glutamic acid, D-arginine, D-cystelne, D-aspartic acid, D-histidine, D-isoleucine, D-proline, 35 D-lysine, D-serine, D-tyrosine and D-valine. Functional groups in these amino acids such as hydroxyl, mercapto, amino and carboxyl groups can be substituted by lower alkyl, benzyl and lower alkanoyl groups. Preferred examples for A include glycine, 8202472 I | J? * -2 * 22547 / Vk / mb sarcosine, D-alanine, D-leucine, D-methionine, D-glutamine, D-phenyl-alanine, D-tryptophan and D-phenylglycine. The group -CH2C (CH2) HCO- in structural formula 1 has an asymmetric carbon atom, so that the group may have the D-form, L-form or DL-form, i.e. a mixture of the 5 D- and L-form. Any mentioned form is part of the invention and the D and DL forms are preferred because of their biological activity.

The proline residue of formula 2, listed on the formula sheet and present in formula 1, may be in the D-form, L-form or a DL-form and any form is included in the present invention, wherein the L- and DL- form are preferred because of their biological activity.

Examples of pharmaceutically acceptable salts of the proline derivatives of formula 1 include the salts with alkali metals such as sodium and potassium, salts with alkaline earth metals such as calcium and magnefeiura and salts with basic amino acids such as arginine and lysine. Calcium salts and lysine salts are particularly preferred.

The present invention also relates to a process for preparing proline derivatives of formula 1 or pharmaceutically acceptable salts thereof. The compounds of formula 1 are prepared by carrying out a method in which a compound of formula R-A'-SC ^ CCCH ^ JHCOOH, wherein A ', is glycine, sarcosine or a protected or unprotected or -D-amino acid residue and wherein R has the same meaning as indicated above, whether a reactive derivative thereof is reacted with a compound of formula 3 listed on the formula sheet, wherein R 'is a hydrogen atom or a carboxyl protecting group, or a reactive derivative thereof, wherein the protecting group is removed from the reaction product and the product is optionally converted into a pharmaceutically acceptable salt.

If A in formula 1 has a free functional group that does not participate in the reaction such as a mercapto, hydroxyl, amino or carboxyl group, the compound of formula R-A'-S-CHgCCCH-HCOOH is a derivative in which such a group may be protected .

Any functional group in which A * does not participate in the reaction can be protected by those groups commonly used in peptide synthesis and which can be removed under relatively mild conditions. For example, a mercapto group can be protected by an aralkyl group such as trityl, benzyl or p-methoxybenzyl, a hydroxyl group can be protected by a benzyl group; an amino group 8202472, * '* j »c -3- - 22547 / Vk / mb.

can be protected by a t-butyloxycarbonyl group and a carboxyl group can be protected by a t-butyl group. These protecting groups can be removed by treatment with hydrogen fluoride, trifluoroacetic acid or hydrogen chloride. On the other hand, a carboxyl-5 group is protected by a lower alkyl-substituted silyl group such as trimethylsilyl group which can be removed by treatment with water. When R * in Formula 3 is a carboxyl protecting group, it may be a t-butyl group or a lower alkyl-substituted silyl group and may be removed by treatment with hydrogen fluoride, trifluoroacetic acid, hydrogen chloride or water.

The reactive derivatives of the compound of formula R-Af-S-CHgC (CH2) HC00H are such that the oarboxyl group that reacts is activated. The carboxyl group can be activated into any of the forms commonly employed in peptide synthesis such as activated amides, acid halides, activated esters and mixed acid anhydrides. Of these forms, the activated esters with N-hydroxy succinic imide, mixed acid anhydrides with carboxylic acid monoesters and activated amides with carbonyldiimidazole are preferred. Carbodiimides such as dicyclohexyloarbodiimide can be used as a condensing agent to form an amide bond between the oarboxyl group and the imino group.

The reactive derivative of the compound of formula 3 is such that the imino group of the compound is activated. The imino group can be activated by any of the methods commonly employed in peptide synthesis, such as introducing a silyl group such as trimethylsilyl group, the "phosphazo method" using a phosphorus compound such as phosphorus trichloride (see Ann. Chem., 572, 96-). 1951), the phosphoric ester method using a phosphoric ester such as tetraethyl ester of pyrophosphoric acid or the "N-carboxy anhydride method" (NCA method).

The reaction can be carried out in an inert organic eplosing agent such as tetrahydrofuran, dioxane, dimethylformamide, hexamethylphosphotriamide, chloroform, dichloromethane or acetonitrile. The reaction is generally run under cooling or at ambient temperature (-50 to 20 ° C), preferably between -30 and 10 ° C. The reaction time varies with the reaction temperature, the compounds that are reacted with each other and the solvent and the reaction time is usually between 0.5 and 48 hours, preferably between 1 and 6 hours.

8202472 -4- 22547 / Vk / mb

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After the amde-forming reaction, the protecting groups in the reaction product are removed by any of the methods mentioned above for the respective protecting groups.

The final compound can be separated from the reaction mixture and purified using a known method, for example, various chromatographic operations using silica gel, dextran cross-linked polymer and porous polymers such as styrene-divinylbenzene or acrylic acid esters. A suitable solvent developing agent can be selected from chloroform, ethyl acetate, methanol, ethanol, tetrahydrofuran, benzene, water and acetonitrile. Alternatively, the final compound can be obtained by first separating the reaction product in the form of an organic salt such as a dicyclohexyl amine salt which can then be treated with an acid such as hydrochloric acid or potassium hydrogen sulfate.

The compound of formula 1 thus obtained has a carboxyl group on the proline side, so that it is able to form salts with various basic substances if desired. Salts with pharmaceutically acceptable basic substances are of particular interest. Such salts can be prepared using a known method of treating the carboxyl group with an equimolar amount of any of the above bases such as alkali metals, alkaline earth metals and basic amino acids.

The compound of formula R-A1-S-CH2C (CH2) HCOOH used as starting material in the method of the invention can be easily prepared by using the following method.

A compound of formula R-OH, where R has the same meaning as indicated above, or a reactive derivative thereof in the carboxyl group is reacted with an amino acid of formula H2N-Att-C00RM, where AM is an amino acid residue equivalent to A minus the group NH or CsO, R "is a hydrogen atom or a carboxyl protecting group, or with a reactive derivative thereof, followed by removal of RM, If this is a protecting group to form a compound of formula R-NH- AM-C00H, where R and A "have the same meaning as indicated above, after which the compound of formula 35 R-NH-A" -C00H or a reactive derivative thereof is reacted with a compound of formula HS-CH2CS (CH ^ C00R, M, where Rm is hydrogen or a carboxyl protecting group, followed by removal of R, M if it is a protecting group.

Examples of the reactive derivative in the carboxylic acid group 8202472 t-22547 / Vk / mb of the compound of formula R-OH include compounds commonly used in peptide synthesis such as activated amides, acid halides, activated esters and mixed acid anhydrides.

If R, f in the amino acid of the formula ngN-A 'COOR "is a protecting group, examples of examples preferably to be used are a t-butyl group, which can be removed using hydrogen fluoride, trifluoroacetic acid or hydrogen chloride, a benzyl group, which can be removed by catalytic reduction with palladium or the like, and lower alkyl groups such as methyl and ethyl (which can be removed by hydrolysis under alkaline conditions).

The reactive derivative of the amino acid of formula C00RM is such that the amino group is activated and this derivative is prepared by one of the activation operations commonly employed in peptide synthesis such as the introduction of a silyl group (such as trimethylsilyl group), the "phosphazomethod", the phosphoric acid ester method and the N-carboxy anhydride method.

An oarbodiimide such as dicyclohexylcarbodiimide can be used as a condensing agent to form an amide bond between the compound of formula R-OH and the amino acid of formula H 1 HA 2 -COOR ". The reaction can be carried out in an inert organic solvent such as tetrahydrofuran , dioxane, dimethylformamide, hexamethylphosphoric triamide, chloroform, dichloromethane or acetonitrile The reaction is generally carried out under cooling or at room temperature (-50 to 20 ° C), preferably between -30 and 10 ° C. The reaction period varies with the reaction temperature, the compounds which are reacted with each other in the solvent and are generally between 0.5 and 48 hours, preferably between 1 and 6 hours.

If RM in formula RgN-A '^ COOR1' is a protecting group, it can be removed from the reaction product by any of the known methods described above.

The compound of formula R-NH-AH-C00H can be separated from the reaction mixture and purified by a known method, for example, by recrystallization from ethyl acetate, n-hexane, acetone or water or by various chromatographic operations as described for separation and purifying the compound of formula 1, or by decomposing an organic salt of the reaction product with an acid.

The compound of formula R-NH-AM-C00H can be reacted with the compound of formula Ηε-ΟΗ ^ Η ^ Η ^ ΟΟΟΗ '", by the 8202472' *» · (* -6- 22547 / Vk / mb reaction of the carboxyl group in the first compound with the mercapto group in the last compound in the presence of a condensing agent, namely carbodiimides such as dicyclohexylcarbodiimide A reactive derivative of the compound of formula R-NH-A "-C00H can be reacted with a compound of formula HS-CH 1 CH 1 CH 2) COOR ". Examples of the reactive derivative of the compound of formula R-NH-A" -C00H include activated amides, acid halides, activated esters and mixed acid anhydrides. Activated amides with carbonyl diimidazole are preferred. The carboxyl group in the compound of formula HS-CH8-CHCCH2 -C00R, M may or may not be protected. The preferred protecting groups are t-butyl and other groups which can be removed under acidic conditions.

The thiol ester forming reaction is performed in an inert organic solvent such as tetrahydrofuran, dioxane, diraethylformaraide, hexamethylphosphotriamide, chloroform, dichloromethane and acetonitrile.

The reaction is usually carried out under cooling or at ambient temperature (-50 to 20 ° C), preferably between -30 and 10 ° C when the compound of formula R-NH-A "-C00H is used in the form of a Activated amide or acid halide and at a temperature between -10 and 10 ° C when the compound is used in the form of an acid anhydride The reaction period varies depending on the reaction temperature, the compounds being reacted with each other and the solvent and is usually between 0.5 and 48 hours, preferably between 1 and 6 hours After the thiol ester reaction, R, M is removed from the reaction product if it is a protecting group If the protecting group is a t-butyl group it may are removed according to a known method, i.e. by reaction with hydrogen fluoride, trifluoroacetic acid or hydrogen chloride.

The final compound can be separated from the reaction mixture and purified by a known method such as recrystallization from an organic solvent such as ethyl acetate or n-hexane, various chromatographic operations as described in connection with the separation and purification of the compound of formula 1, or by decomposition of an organic salt of the reaction product with an acid.

Another process for the preparation of the final compound according to the invention consists of a reaction between a compound of formula R-A'-OH, wherein R and A 'have the same meaning as above 8202472 ..- »y * * · -7 - 22547 / Vk / mb, or a reactive derivative thereof with a compound of formula 4 listed on the formula sheet, where Rf has the same meaning as indicated above, removing a possible protecting group from the reaction product and optionally converting the product 5 into a pharmaceutically acceptable salt.

If A in form 1 has a free funtone group that does not participate in the reaction such as a meroapto, hydroxyl, amino or carboxy group, the compound of form R-A'-OH means a derivative in which such group may be protected.

A possible functional group in A * which does not take part in the reaction deeX is protected by such groups which are known in the peptide synthesis and which can be removed under relatively minimal conditions. For example, a mercapto group can be protected by an araikyi group such as trityi, benzyi or p-methoxybenzyl group, a hydroxy group can be protected by a benzyi group, an amino group can be protected by a t-butyloxicarbonyl group, and a carboxyX group can be protected by a t-butyl group. These groups can be removed by treatment with hydrogen fluoride or hydrogen chloride. On the other hand, a carboxyl group 20 is protected by a lower aikyi-substituted siiyi group such as tri-methylsilyl group which can be removed by treatment with water. When R * in Form 5 is a carboxy protecting group, it may be a t-butyl group or a lower aikyi-substituted siiyi group and may be removed by treatment with hydrogen fluoride, trifluoroacetic acid, hydrogen chloride or water.

The reactive derivative of the compound of form R-A'-OH is such that the carboxyl group that reacts is activated.

The carboxyl group can be activated in a suitable form such as activated amides, acid halides, activated esters or mixed acid anhydrides. Activated amides with carbonyldiimidazole are preferred. Carbodiimides such as dicyelohexylcarbodiimide can be used as a condensing agent to form an amide bond from the carboxyl group and the mercapto group.

The reaction between the compound of formulas R-A'-OH or its reactive derivative and the compound of formulas 4 listed on the formula sheet is preferably carried out in an inert organic solvent such as tetrahydrofuran, dioxane, dimethylformamide, hexamethylphosphotriamide , chloroform, dichloromethane or acetonitrile. The reaction 8202472 -8-22547 / Vk / mb is generally carried out under cooling or at ambient temperature (-50 to 20 ° C) and preferably between -30 and 10 ° C, when the compound of formula RA * -0H is used in the form of an activated amide or acid halide and between -10 and 10 ° C when the compound 5 is used in the form of an acid anhydride. The reaction time varies depending on the reaction temperature, the reacted compounds and the solvent and is usually 0.5-8 hours, preferably between 1 and 6 hours. After the thiol ester forming reaction, possible protecting groups are removed from the reaction product according to one of the methods described for the respective protecting groups.

The final compound can be separated from the reaction mixture and purified by any of the known methods described above, such as various chromatographic operations using silica gel, dextran cross-linked polymer and porous polymers such as styrene divinyl benzene or an acrylic acid ester. A suitable developing solvent can be selected from chloroform, ethyl acetate, methanol, ethanol, tetrahydrofuran, benzene, water and acetonitrile. Alternatively, the final product can be obtained by first separating the reaction product in the form of an organic salt such as dicyclohexylamine salt which can then be treated with an acid such as hydrochloric acid or potassium hydrogen sulfate.

The end compounds according to the invention, namely the proline derivatives of formula 1 and the pharmaceutically acceptable salts thereof, prevent the development of angiotensinell from angiotensin I by inhibiting the activity of the angiotensin converting enzyme. Therefore, they are suitable for the treatment of hypertension caused by angiotensin II and as a means of treating cardiac insufficiency.

The activity of some of the end compounds of the invention to inhibit the angiotensin converting enzyme was measured.

1) The method to be applied was the following. An angiotensin converting enzyme was extracted from a rabbit lung. A 0.111M boric acid-NagCO2 buffer solution (pH = 8.3, 0.6 ml), 0.2 ml of 0.111M boric acid-Na2 CO2 buffer solution (pH = 8.3) with 25 mM benzoyl-35 glycylhistidylleucine (substrate ) and 0.1 ml 0.111M boric-Na 2 CO buffer solution (pH 8.3) with 10 to 10 M of the test compound (13 final compounds of the invention, listed in Table A) were placed in test tubes done and stored therein for 5 to 10 minutes 8202472

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-9- 22547 / Vk / mb I 1 at a temperature of 37 ° C. Then, 0.1 ml of a solution of the enzyme (aetone powder) was added to each test tube and the mixture was stored at a temperature of 37 ° C for 30 minutes. The benzoyl glycyne, obtained with the aid of the enzyme, was extracted with ethyl acetate in the presence of hydrochloric acid and the amount of this was determined by UV absorption at 228 nm. The enzyme activity in the presence of the test compounds relative to that in the absence of the test compound (100) was determined. The concentration of each test compound at which the enzyme's relative activity was 50% was indicated as the activity of each test compound to inhibit the activity of the enzyme and was indicated as an IQ value.

2) The results are shown in Table A.

15 TABLE A

examined ^. * I_n ~ value connection '1 1.6 x ΙΟ'7 2 4.8 x 10'7 20 3 1.3 x ΙΟ'7 4 4.0 x 10'7 5 1.8 x ΙΟ "7 6 2.4 x 10 "7 7 1.0 x ΙΟ'7 25 8 1.5 x ΙΟ" 7 9 1.7 x ΙΟ'7 10 4.8 x 10-7 11 1.7 x 10 “7 12 5.4 x ΙΟ ”7 30 13 4.4 x ΙΟ” 7 * 1)

Note compound 1 = H- [3- (N-oyolohexanecarbonyl-D-alanylthio) -2-D-methylpropan-noyl] -L-proline, compound 2 = N- [3- (N-cyelopropanecarbonyl-D-alanylthio) -2-D-methyl-propanoyl3-L-proline, compound 3 = N- [3- (N-cyclohexanecarbonylglycylthio) -2-D-methylpropanoyl] -L-proline, 8202472 • », / 'I' -10 - 225 ^ 7 / Vk / mb compound 4 = N- [3- (N-cyelohexanecarbonyl-N-methylglycylthio) -2-D-methylpropanoyl-L-proline, compound 5 = N- [3- (N-cyclopropanecarbonylglycylthio) - 2-D-methyl-propanoyl] -L-proline, 5 compound 6 s N- [3- (N-adamantanecarbonylglycylthio) -2-methylpropanoyl-L-proline, compound 7 * N- [3- (N-cyclohexanecarbonyl-D) phenylalanylthio) -2-D-methylpropanoyl) -L-proline, compound 8 = N- [3- (N-cyclohexanecarbonyl-D-leucylthio) -2-D-methyl-10-propanoyl-L-proline, compound 9 = N - [3- (N-cyclohexanecarbonyl-D-tryptophylthio) -2-D-methylpropanoyl-L-proline, compound 10 = N- [3- (N-cyclohexanecarbonyl-D-phenylglycylthio) -2-D-methylpropanoyl] -L- proline, compound 11 = N- [3- (N-cyc lohexanecarbonyl-D-methionylthio) -2-D-methylpropanoyl-L-proline, compound 12 = N- [3- {N-cyclohexanecarbonyl-D-glutaminylthio) -2-D-methylpropanoyl} -L-proline and compound 13 = N - [3- (N-adamantanecarbonyl-D-alanylthio) -2-D-methyl-20-propanoyl-L-proline.

The activity of the end compounds according to the invention has a longer period of time than of known compounds with a similar activity, namely 3-mercapto-2-D-methylpropanoyl-L-proline (known as Captopril), so that the desired control of blood pressure can be obtained with fewer daily dosages.

Captopril and other known compounds cause a sudden drop in blood pressure at the initial period of administration and this can lead to postural hypotensive asthenia {[Lancet, vol. 1, no. 8, 115, biz. 557 C10, 1979) J, but the end compounds of the invention have only a mild hypotensive action during the initial period of administration and only a slight chance of development into orthostatic hypotensive asthenia. Medicines such as Captopril that have a free mercapto group cause various side effects attributed to the mercapto group. Some of these drawbacks are deterioration of taste, urinary albumin production, agranulocytosis and dermal diseases accompanied by fever are reported in Lancet, Vol. 1, no 8160, biz. 150 (January 19, 1980), and Vol, 2, no. 8186, biz. 129 (July 19, 1980) 8202472 "* V ..

-11- 22547 / Vk / mb and South African Medical Journal, vol. 58, 172 (1980). On the other hand, the thioester bond in the final products of the invention is not subject to hydrolysis in vivo and thus little subject to the preparation of a mercapto group. Therefore, in the end compounds 5 of the invention, there is little chance of causing the above-mentioned side effects attributed to the mercapto group,

The proline derivatives of formula 1 and the pharmaceutically acceptable salts thereof can be formulated into compositions for oral administration such as tablets, capsules, granules, powders, syrups and elixir or into a sterile solution or suspension for parenteral administration. For this purpose, the pharmaceutical compositions can be prepared from 4 and more of the end compounds of the invention as active substances and pharmaceutically acceptable auxiliary substances can be added such as fillers, carriers, binders, stabilizers and flavors. The daily dosage of the final compounds of the invention for adults is 0.5 mg to 2 g, preferably 1 to 500 mg for oral administration and 0.1 to 600 mg, preferably 0.3 to 300 mg for parenteral administration.

The process for the preparation of the final compounds according to the invention is further elucidated by means of the following examples, which should not be regarded as limiting.

Example I

The preparation of N-substituted amino acids.

a) With stirring, 4.5 g of D-alanine was dissolved in 230 ml of 1N 25 aqueous Na 2 CO 2. To the solution, 100 ml of tetrahydrofuran with 9.0 g of cyclohexane carbonyl chloride was added dropwise at 5-10 ° C, at which temperature the mixture was stirred for 30 minutes, then stirred at room temperature for 1.5 hours. Then 2N-HCl was added to the reaction mixture to adjust its pH between 1 and 2. The reaction mixture was extracted with ethyl acetate and the organic layer was washed with a saturated NaCl solution and dried over magnesium sulfate. The filtrate was evaporated under reduced pressure to obtain a crude compound. Recrystallization from ethyl acetate / n-hexane gave 4.65 g of N-cyclohexanecarbonyl-D-35 alanine of which the (e ^ value was +26.6%).

b) An ester of adamantane carboxylic acid and N-hydroxy succinic acid imide was dissolved in 30 ml of tetrahydrofuran in an amount of 3.0 g. To the solution, 5 ml of water with 0.89 g of D-alanine and 8202472-122547 / Vk / mb ft, 1.1 g of triethylamine was added and the mixture was stirred at a temperature of 5 ° C overnight. After removing tetrahydrofuran, water was added to the residue and 2N-HCl was further added to the mixture to adjust its pH to between 5 1 and 2. The mixture was then treated as mentioned under a) above. - obtaining 0.38 g of N-adamantane carbony1-D-alanine. The H value is + 11.6 °, measured in methanol at an concentration of 1.0. (C = 1.0 MeOH).

The compounds listed in Table B were prepared by performing one of the methods a) or b) as listed in Table B.

Water, tetrahydrofuran was used as the solvent for carrying out the reaction.

TABLE B

N-substituted | | reaotant physical properties of amino acids ". 1 - 7 N-hydroxybar-1 H-NMR (CDqOD,) 15 N-adamantane-carbo- (b) -atanoic acid dioyalo-1.6-2.3 ≤ 5H, m), nyigiycme. hexylcarbodiimide 3.86 (2H, S) N-cyclohexanecar-, * cyclohexanecarboxylic acid Q1 * 1 R0 * 1) bonyl-D-phenylalanine nyl chloride D 'N-cyclohexanecyclohexanecarbon-1q qo * 1) bonyl-D-leucine Ca' nyl ') Y' 20 N-cyclohexane car-. »Cyclohexane carbon .-, 1f) _0 * 1) bonyl-D-tryptophan 1 'nyl chloride" "N-cyclohexane car-, * cyclohexane carbon.-J _ _0 # ΐ) bonyiD-phenylglycine ^ nyl chloride L * N -Cyclohexane -carbo-. * cyclohexane -carbo- "0.1" -Nyl-D-methionine-methyl chloride JD "" "N-cyclohexane-carbon. . cyclohexanecarbo- ς -ό * 1) nyl-D-glutamine '' 'nyl chloride LT) "+ * N-cyclopropanecarbo-. cyclopropanecar- ΓρΠ _ ς1 -0 * 1) nyl-D-alanine' 'bonyl chloride Ί)" + 5 'N-cyclohexane carbon. . cyclohexane carbon-1 H NMR (CDoDD, S) nylglyoin nyl chloride * 2 m 3, 3.95 (2H, S) 1 H NMR (CD3OD, *) N-cyclohexane carbon. . cyclohexane carbonylsaroosin (a) nYX chloride (M) (13H5) (2H, two S) 1 H NMR (CDoOD, 6) N-cyclopropane carbon. - cyolopropane carbon- 0.5-2.1 (4H, m), nylglyoin 'a' nyl chloride 2.4-2.9 (1H, m), 3.90 (2H, S)

Note: The measurement was performed in methanol at a concentration of 1.0 (C = 1.0; MeOH).

8202472. * * * 1 * -13- 22547 / Vk / mb

Example II

a) 3-- CN-cyelohexane -earbonyl-D-alanylthio) -2-D-methyl-ethyl-acetic acid.

5.97 g of N-cyclohexane-5-carbonyl-D-alanine was dissolved in 100 ml of dry tetrahydrofuran. 5.84 g of carbonyldi-Imldazole were added to the solution at a temperature between -20 and -15 ° C and the mixture was stirred at this temperature for one hour. Thereafter, 3.60 g of 3-mercapto-2-D-methyl-ethano-carboxylic acid was added and the mixture was stirred at a temperature between -15 and -10 -10 ° C for one hour and then at room temperature for another hour. The mixture was evaporated under reduced pressure to remove the solvent. 40 ml of water were added to the residue and 2N-HCl was added to the mixture to adjust its pH to between 1 and 2. The mixture was extracted with ethyl acetate and the organic layer was washed twice with a saturated NaCl solution and dried over magnesium sulfate. The filtrate was evaporated under reduced pressure to obtain a crude compound. Recrystallization from ethyl acetate / n-hexane gave a colorless prism-like crystal of the title compound in an amount of 7.50 g, corresponding to a yield of 83¾. The melting range is 149-152 ° C.

The [οζΐρ value * + 46.4 ° (C = 1.07, MeOH), the NMR spectrum (CD ^ OD, ^) gives the following values: 1.20 (3H, d), 1.35 ( 3H, d), 1.20-2.0 (11H, m), 2.40-2.80 (1H, m), 3.05 (2H, m) and 4.50 (1H, m).

B) 3- (N-cyclohexanecarbonyl-D-phenylglycylthio) -2-D-methyl-ethanecarboxylic acid.

The procedure mentioned under a) was repeated using 2.61 g of N-cyelohexanecarbonyl-D-phenylglycine, 1.95 g of carbonyldimidazole and 1.20 g of 3-meroapto-2-D-methyl-ethanecarboxylic acid. Thus, a yellow oil of the title compound was obtained in an amount of 2.07 g, corresponding to a yield of 57¾.

The [o ^ value = -48.7 ° (C = 1.19, MeOH).

The NMR spectrum (ΟϋΟΙ ^, ί) gives the following values: 1.24 35 (3H, d), 1.20-2.10 (11H, m), 2.40-2.80 (1H, m) 3.10 (2H, m), 5.75 (1H, d), 6.40-6.80 (1H, m), 7.50 (5H, S) and 8.90 (1H, S).

o) 3- (N-cyclohexanecarbonyl-D-leucylthio) -2-D-methyl-ethylcarboxylic acid.

8202472 t -14- 22547 / Vk / mb

The procedure mentioned under a) was repeated using 2.41 g of N-cyclohexanecarbonyl-D-leucine, 1.95 g of carbonyldiimidazole and 1.20 g of 3-mercapto-2-D-methyl-ethanecarboxylic acid. An oil of the title compound was obtained in an amount of 2.40 g, corresponding to a yield of 70%.

The CeQp value is + 12.2 ° (C = 1.03, MeOH). The data from the NMR analysis (CDCl 3, &) is 0.95 (6H, d), 1.28 (3H, d) , 1.20-2.10 (11H, m), 2.40-2.80 (1H, m), 3.10 (2H, m), 4.70 (1H, m), 6.00-6 .30 (1H, m) and 9.20 (1H, S).

D) 3- (N-cyclohexane carbonyl-D-glutaminylthio) -2-D-methylethane carboxylic acid.

The procedure mentioned under a) was repeated using 2.56 g of N-cyclotiexanecarbonyl-D-glutamine, 1.95 g of carbonyldiimidazole and 1.20 g of 3-mercapto-2-D-methyl-ethanecarboxylic acid. A sample in an amount of 0.70 g of the title compound was prepared, corresponding to a yield of 20 µ. The melting range was 146-149 ° C.

The DOip value s + 10.8 ° (C = 1.05if MeOH). The data from the NMR analysis (CD30D, £) are: 1.20 (3H, d), 1.20-2.10 (11H, m), 2.30 (4H, m), 2.40-2 .80 (1H, m), 3.05 (2H, m) and 4.50 (1H, m).

E) 3- (N-cyclopropanecarbonylglycylthio) -2-D-methyl-ethanecarboxylic acid.

The procedure mentioned under a) was repeated using 0.78 g of N-cyclopropanecarbonylglycine, 1.06 g of carbonyldiimidazole and 0.66 g of 3-mercapto-2-D-methyl-ethanecarboxylic acid. An oil of the title compound was prepared in an amount of 0.70 g, corresponding to 52% yield.

The {bQjj value -33.8 ° (C = 1.01, MeOH).

The data from the NMR analysis (CDCl 3, S) are: 0.70-1.20 (4h, m), 1.25 (3H, d), 1.30-1.80 (1H, m), 2.40-2.80 (1H, m), 3.14 (2H, m), 4.20 (2H, d) and 7.30 (1H, S).

f) 3- (N-adamantanecarbonylglycylthio) -2-D-methyl-ethanecarboxylic acid.

The procedure mentioned under a) was repeated using 2.38 g of N-adamantanecarbonylglycine, 1.95 g of carbonyldiimidazole and 1.20 g of 3-mercapto-2-D-methyl-ethanecarboxylic acid. A sample was obtained in an amount of 1.08 g of the title compound, corresponding to a yield of 32%.

The melting point was 132 ° C (decomposition) and the [44d] value is -8.3 ° (C = 1.01, MeOH).

8202472 ·· -. ·. * * -15- 22547 / Vk / mb

The data from the NMR analysis (CD ^ OD, ξ) are: 1.25 (3H, d), 1.65-2.30 (15H, m), 2.40-2.80 (1H, m) , 3.14 (2H, m) and 4.06 (2H, d).

g) 3- (N-cyclohexanecarbonyl-D-phenylalanylthio) -2-D-methyl-ethanecarboxylic acid.

The procedure mentioned under a) was repeated using 1.37 g of N-cyclohexanecarbonyl-D-phenylalanine, 1.0 g of carbonyldimidazole and 0.60 g of 3-mercapto-2-D-methyl-ethano-carboxylic acid. An oil of the title compound was obtained in an amount of 1.5 g, corresponding to a yield of 79/6.

The C <0 D value = + 13.9 ° (C = 1.02, MeOH).

The data from the NMR analysis (CDCl 3, S) are: 1.25 (3H, d), 1.20-2.0 (10H, m), 2.1-3.0 (2H, m), 3.0-3.4 (4H, m), 4.95 (1H, t), 6.19 (1H, d), 7.24 (5H, S) and 10.12 (1H, S).

h) 3- (N-cyclohexanecarbonyl-D-methylthio) -2-D-raethylethane carboxylic acid. ^

The procedure mentioned under a) was repeated using 0.325 g of N-cyclohexanecarbonyl-D-methionine, 0.25 g of carbonyldimidazole and 0.15 g of 3-mercapto-2-D-methylethanecarboxylic acid. An oil of the title compound was prepared in an amount of 0.29 g, corresponding to a yield of 64 held.

The C <3 D value = -7.9 ° (C = 1.0, MeOH)

The data from the NMR analysis (CDCl ^, £) are: 1.26 (3H, d), 1.2-2.0 (10H, m), 2.09 (3H, S), 2.0- 3.2 (8H, m), 4.5-5.0 (1H, m), 6.62 (1H, d) and 10.07 (1H, S).

I) 3- (N-cyclohexanecarbonyl-D-tryptophylthio) -2-D-methylethanecarboxylic acid.

The procedure mentioned under a) was repeated using 0.157 g of N-cyclohexanecarbonyl-D-tryptophan, 0.1 g of carbonyldiimidazole and 0.06 g of 3-mercapto-2-D-methyl-ethanecarboxylic acid.

A gummy sample of the title compound was obtained in an amount of 0.17 g, corresponding to a yield of 8256.

The t <3D value s -7.1 ° (c s 1.0, MeOH).

The data from NMR analysis (CDCl ^ nT) are: 1.26 (3H, d), 1.2-2.4 (11H, m), 2.5-2.9 C1H, m), 3 .0-3.3 (2H, m), 3.35 (2H, d), 4.87 (1H, t), 6.4-6.9 (2H, broad S), 6.9-7, 8 (5H, m) and 10.5 (1H, S).

Example III

a) N- [3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methyl-propanoyl] -L-proline. β 2 0 2 4 7 2, «-16- 22547 / Vk / mb

1.51 g of 3- (N-cyclohexane-carbonyl-D-alanylthio) -2-D-methyl-ethanecarboxylic acid were dissolved in 40 ml of dry tetrahydrofuran. 0.61 g of trigthylaraine and 0.65 g of ethyl chloroformate were added to the solution with stirring at a temperature of -5 ° C. Five minutes later, a solution containing 0.58 g of L-proline and 0.61 g of triSthylamine dissolved in 5 ml of water was added and the mixture was stirred at 0 ° C for an hour, then at room temperature for 30 minutes.

The mixture was evaporated under reduced pressure to remove the solvent. After adding water to the residue, 2N-HCl was added to the mixture to adjust the pH value from 1 to 2. The reaction mixture was extracted with ethyl acetate and the organic layer was washed twice with saturated NaCl. solution and dried over magnesia sulfate. The filtrate was evaporated under reduced pressure to remove ethyl acetate and the residue was purified chromatographically on silica gel using a mixture of chloroform and methanol (100: 1 to 100: 2) as an eluent. The fractions containing the final product were collected and evaporated under reduced pressure to obtain a gummy sample in an amount of 0.3 g of the title compound.

H) The same compound could be prepared using the following procedure. 6.03 g of 3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methyl-ethanecarboxylic acid and 2.3 g of N-hydroxysuccinimide were dissolved in 50 ml of dry tetrahydrofuran. 4.3 g of dicyclohexylcarbodiimide were added to the solution at a temperature between 0 and 5 ° C and the mixture was stirred overnight at this temperature. After the reaction, the precipitate was filtered off and the insoluble matter was washed with a small amount of tetrahydrofuran. The filtrate and washings were combined and evaporated under reduced pressure. Ethyl acetate was added to the residue and the mixture was filtered. The ethyl acetate solution was washed with 0.5N HCl, water, aqueous Na 2 pCO 2 solution and saturated NaCl solution in the order listed. The solution was dried over magnesium sulfate, evaporated under reduced pressure and the residue solidified after adding a mixture of ethyl acetate and hexane (1:10).

The yield of solid product was 6.90 g, corresponding to 87? yield. The melting range was 113-116 ° C.

The [<* 3D ~ value = + 14.2 ° (C = 1.05, MeOH).

8202472 The data from NMR analysis (CDCl ^ i) are: 1.38 (6H, d), T ti -17- 22547 / Vk / mb 1.20-2.20 (11H, in), 2.82 (4H, S), 2.8-3.30 (3H, m), 4.75 (1H, m) and 6.30 (1H, broad d).

3.98 g of an activated ester was obtained in 40 ml of tetrahydrofuran. 5 ml of water with 1.15 g of L-proline 5 and 1.26 ml of N-ethylmorpholine were added to the solution and the mixture was stirred overnight. The mixture was evaporated under reduced pressure to remove the solvents and water was added to the residue. 2N-HCl was added to the mixture to adjust the pH to a value between 1 and 2. The reaction mixture was extracted with ethyl 10 acetate and the organic layer was washed with a saturated NaCl solution and dried over magnesium sulfate. The filtrate was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography using a mixture of chloroform and methanol (100: 1 to 100: 2) as an eluent. The product was obtained in an amount of 1.45 g by evaporation of the eluate under reduced pressure and this substance was found to be equal to the title compound on the basis of the data from the NMR analysis.

c) The title compound could also be prepared using the following procedure.

4.52 g of 3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methyl-ethanecarboxylic acid were dissolved in 5 ml of thionyl chloride. A drop of dimethylformamide was added to the solution and the mixture was stirred at room temperature overnight. The mixture was evaporated under reduced pressure to remove the solvents and 2 ml of dry toluene was added to the residue and the mixture was again evaporated under reduced pressure to remove the solvent. 10 ml of dry tetrahydrofuran were added to the residue with stirring.

The tetrahydrofuran-containing solution containing the acid chloride was added dropwise at a temperature between 5 and 10 ° C below.

Stirring to 25 ml of water with 2.3 g of L-proline and 2.5 g of NagCO 4 dissolved therein. The mixture was stirred at this temperature for one hour and at ambient temperature for another two hours. The reaction mixture was subjected to an extraction with chloroform. 2N-HCl was added to the aqueous layer to adjust its pH to a value between 1 and 2. After extraction with chloroform, the chloroform layer was washed twice with a saturated NaCl solution and dried over magnesium sulfate. After removing chloroform, the residue was subjected to column chromatographic purification on silica gel using 8202472 t -18-22547 / Vk / rab using a mixture of chloroform and methanol (100: 1 to 100: 2) as an eluent. After evaporation under reduced pressure, 1.6 g of a gummy substance was obtained. This substance was found to be the same as the title compound based on the NMR analysis.

D) The compound mentioned in the preamble can also be prepared by using a following method. 3.01 g of 3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methyl-ethanecarboxylic acid and 1.71 g of t-butyl ester of L-proline were dissolved in 50 ml of dry dichloromethane and 2.15 g were added. dicyclohexylcarbodiimide with stirring and cooling with ice. The mixture was stirred at the same temperature for 30 minutes and left at 5 ° C for one night. The reaction mixture was filtered and the insoluble material was washed with dichloromethane. The filtrate and washings were combined and washed with 1N HCl, water, 1N NaHCO 2 and a saturated NaCl solution in the order listed, dried over MgSO 2 and evaporated under reduced pressure to give a gummy sample of t- butyl ester of N- [3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-mebhylpropanoyl] -L-proline in an amount of 4.3 g. A 4.0 g sample of the ester was dissolved in 30 ml of anisole 20 and 10 ml of tetrafluoroacetic acid was added to the solution. The mixture was stirred at room temperature for one hour and evaporated under reduced pressure to remove the excess trifluoroacetic acid.

The residue was purified by chromatography on silica gel (column: 2 x 35 cm) using a mixture of methanol and chloroform (1: 100 to 3: 100) as an eluent. Fractions with the final compound were collected and evaporated under reduced pressure to obtain 3.4 g of gummy substance. This material was found to be the same as the title compound based on NMR analysis and thin layer chromatography.

The reactants, solvents used in the reaction and the methods used to prepare the title compound in Example III are listed in Table C, together with the physical properties. Other compounds were prepared using one of the methods described in Examples III a) to d) and details of the particular method are set forth in Table C together with the data of the physical properties of the final compounds. In Table C, under Rf value, it is stated that * 1) the Rf value refers to a value obtained by thin layer 8202472, * * -t9- 22547 / Vk / mb chromatography using CHCl 2 / MeOH / AcOH ( 2: 1: 0.003) and * 2) refers to the use of the eluent n-BuOH / AcOH / H 2 O (4: 1: 1).

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Example IV

Various salts were formed from the samples of N- (3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methylpropanol] -L-proline prepared in Examples III and V.

5 a) Ca salt.

A 3.98 g sample of the compound prepared according to Example III or V was dissolved in 40 ml of methanol. 0.84 g of calcium acetate hydrate was added to the solution and the mixture was heated to reflux for one hour. The insoluble matter 10 was filtered off and the filtrate was evaporated under reduced pressure. Chloroform was added to the residue and the mixture was filtered and evaporated under reduced pressure. Diethyl ether was added to the residue and the mixture was filtered and dried with air to give a calcium salt of the compound of Example III or V in an amount of 3.40 g. The D * ·! D value = -47.2 ° (C = 1.0; MeOH).

b) Mg salt.

The procedure mentioned under a) was repeated using 3.09 g of the compound prepared according to example III or V and 0.772 g of magnesium acetate tetrahydrate. A magnesium salt of the compound of Example III or V was prepared in an amount of 2.42 g. The CoOD value = -46.6 (C = 1.0, MeOH).

c) Lysine salt.

The compound prepared in Example III or V was dissolved in 22 ml of methanol in an amount of 1.19 g. 0.416 g of lysine was added to the solution and the mixture was stirred at room temperature for one hour and evaporated under reduced pressure. Chloroform was added to the residue and the mixture was filtered and evaporated under reduced pressure. Dimethyl ether was added to the residue and the product was filtered off under reduced pressure to obtain a lysine salt of the compound of Example III or V in an amount of 1.54 g. The Codj) value = -23.1 ° (C = 1.0, MeOH).

d) Na salt.

2.27 g of the compound prepared in Example III or V were dissolved in 25 ml of methanol. To the solution, 0.514 g of sodium acetate was added and the mixture was stirred at room temperature for 30 minutes and evaporated under reduced pressure. To the residue was added 200 ml of a mixture of methanol and chloroform (3: 1 volume 8202472-252547 / Vk / ts ratio) and the resulting mixture was filtered and evaporated under reduced pressure. The residue was dissolved in methanol and the solution was filtered and evaporated under reduced pressure. Ethyl acetate was added to the residue and the product was filtered off under suction to give a sodium salt of the compound of Example III or V in an amount of 1.85 g. Det * S ^ value = -26.7 ° (C = 1.0, MeOH).

e) Dicyclohexylamine salt.

The compound of Example III or V was dissolved in 120 ml of acetonitrile in an amount of 13.1 g. To the solution was added 6 ml of dicyclo-hexylamine with stirring. The mixture was further stirred for 30 minutes and left overnight. The precipitate was filtered under reduced pressure and air dried. The crude crystalline substance obtained was suspended in 300 ml of acetonitrile and the suspension was heated to reflux for 30 minutes.

The slurry was cooled and the crystal filtered under reduced pressure and air dried to give a dicyclohexylamine salt of the compound of Example III or V in an amount of 12.2 g.

Det * Qj) value s -24.5 ° (C s 1.0, MeOH).

Example V.

N-3- (Ni-cyclohexanearbonyl-D-alluylthio) -2-D-methylpropanoyl]} -L-proline.

5.98 g of N-cyclohexane-arbonyl-D-alanine were dissolved in 80 ml of dry tetrahydrofuran. To the solution, 5.84 g of carbonyl diimidazole was added at -18 ° C with stirring, while the mixture was cooled with ice. The mixture was stirred at this temperature for 1 hour and then 6.29 g of N- (3-mercapto-2-D-methylpropanoyl) -L-proline was added to the mixture, followed by stirring at -18 ° C for 30 minutes and then at room temperature for 1 hour. After the completion of the reaction, the mixture was evaporated under reduced pressure to remove the solvent. 50 ml of water and 2N-HCl were added to the mixture to adjust the pH thereof to a value between 1 and 2. The mixture was extracted with ethyl acetate and the ethyl acetate-containing layer was washed with a saturated NaCl solution, dried over MgSO4 and evaporated under reduced pressure. 120 ml of acetonitrile were added to the residue and 6 ml of dicyclohexylamine (DOHA) were then added. The mixture was stirred at ambient temperature for 1 hour. After it was left overnight, the precipitate was filtered off and air dried to obtain 14.35 g of crude DCHA salt. The coarse salt was slurried. -261-22547 / Vk / ts dealt in 300 ml of acetonitrile and the suspension was heated under reflux for 30 minutes. After cooling, the precipitate was collected under suction and air dried to obtain 12.20 g of white DCHA salt. The DCHA salt was suspended in an amount of 12.20 g in 90 ml of ethyl acetate and 60 ml of 0.5 N aqueous KHSO 2 was added to the suspension and the mixture was shaken. The organic layer was washed with distilled water, dried over MgSOjj and evaporated under reduced pressure to obtain 8.64 g of gum. This material was found to be equal to the title compound based on data from 10 NMR analysis and thin layer chromatography.

Example VI.

N-t3- (N-cyclohexanecarbonyl-glycylthio) -2-D-methylpropanoy] J * L-proline.

1.02 g of N-cyclohexanecarbonyl-glyoine was dissolved in 10 ml of dry tetrahydrofuran. To the solution, 20 ml of dry tetrahydrofuran with 1.07 g of carbonyldiimidazole was added at -20 ° C with stirring, while the mixture was cooled with ice. The mixture was stirred at this temperature for 1 hour and then 6 ml of dry tetrahydrofuran was added containing 1.09 g of N- (3-mercapto-2-D-methylpropanoyl) -L-proline followed by stirring at - 20 ° C for 30 minutes and then at ambient temperature for 1 hour. After the completion of the reaction, the mixture was evaporated under reduced pressure to remove the solvent. The residue was purified by column chromatography over silica gel (column: 2 x 35 cm) using a mixture of methanol and chloroform (1: 100 to 3: 100) as an eluent. The fractions containing the final product were combined and evaporated under reduced pressure to obtain 1.16 g of gum. The substance was found to be the same as the title compound based on NMR analysis and thin layer chromatography.

Example VII.

N-t3- (N-cyclopropanecarbonyl-D-alanylthio) -2-D-methylpropanoyl-30-L-proline.

635 mg of N-cyclopropanecarbonyl-D-alanine and 0.70 ml of triethylamine were dissolved in 14 ml of dry tetrahydrofuran. To the solution, 0.48 ml of ethyl clone formate was added at 15 ° C with stirring while the mixture was cooled with ice. The mixture was stirred / at this temperature for 15 minutes and then 1.09 g of N- (3-mercapto-2-D-methylpropanoyl) -L-proline and 10 ml of dry tetrahydrofuran containing 0.70 ml triethylamine added, followed by stirring at: -15 ° C for 15 minutes and then overnight at 5 ° C. After the completion of the reaction, 8202472 ff -27-22547 / Vk / ts the mixture was evaporated under reduced pressure at 30-35 ° C to remove the solvent. To the residue was added 1Q ml of water and 2N-HCl to adjust its pH to a value between 1 and 2. The mixture was extracted with chloroform and the chloroform layer was washed with 5 saturated NaCl solution, dried over MgSO 4 and evaporated under reduced pressure. The residue was subjected to a silica gel column chromatographic purification (column: 2 x 35 cm) using a mixture of methanol and chloroform (1: 100 to 3: 100) as an eluent. The fractions with the final compound were corabinated and evaporated under reduced pressure to give 395 gum-like substance. This material was found to be the same as the title compound based on NMR analysis and thin layer chromatography.

Example VIII.

N- [3- (N-cyclohexanecarbonyl-N-methylglycylthio) -2-D-methylpropanoyl] -15 L-proline.

The procedure in Example 6 was repeated except that N-cyclohexanecarbonylglycine was replaced with 1.09 g of N-cyclohexanecarbonyl-N-methylglycine, 1.09 g of carbonyldiimidazole and 1.09 g of N- (3-mercapto-2- D-methylpropanoyl) -L-proline. Thus, 0.76 g of gummy substance was obtained and found to be equal to the title compound based on NMR analysis and thin layer chromatography.

Example IX.

N-t3- (N-cyclopropanecarbonylglycylthio) -2-D-methylpropanoyl "] -L- proline.

The procedure in Example 6 was repeated except that N-cyclohexanecarbonyl-glycine was replaced by 0.96 g of N-cyclopropanecarbonylglycine, 1.09 g of carbonyldiimidazole and 1.09 g of N- (3-mercapto-2-D- methylpropanoyl) -L-proline. A gummy substance was obtained in an amount of 0.4 g, which was found to be equal to the title compound based on NMR analysis and thin layer chromatography.

Example X.

N-n3- (N-adamantanecarbonylglycylthio) -2-D-methylpropanoyl3 -L-proline.

The procedure in Example 6 was repeated except that N-cyclohexanecarbonylglycine was replaced with 1.42 g of N-adamantanecarbonyl-glycine, 1.09 g of carbonyldiimidazole and 1.09 g of N- (3-mercapto-2-D-methylpropa- noyl) -L-proline. A gum-like substance was prepared in an amount of 1.7 g, which was found to be equal to the title compound 8202472 / * -28-22254 ^ / Vk / ts based on NMR analysis and thin film chromatography.

Example XI.

N-3- (N-cyclohexanecarbonyl-D-phenylalanylthio) -2-B-methylpropanoyl] -L-proline.

The procedure in Example VI was repeated except that N-cyclohexanecarbonylglycine was replaced by 1.5 * 1 g N-cyolohexanecarbonyl-D-phenylalanine, 1.09 g carbonyldiimidazole and 1.09 S N- (3-mercapto 2-D-methylpropanoyl) -L-proline. Thus, 0.97 g of gummy substance was obtained, which was found to be equal to the title mentioned on the basis of NMR analysis and thin layer chromatography.

Example XII.

N-3- (N-cyclohexanecarbonyl-D-leucylthio) -2-D-methylpropanoyl * | - L-proline.

The procedure described in Example 6 was repeated except that 15 N-cyclohexanecarbonylglycine was replaced by 1.20 g of N-cyclohexanecarbonyl-D-leucine, 0.97 g of carbonyldiimidazole and 0.98 g of N- (3-mercapto-2- D-methyl-propanoyl-L-proline. A gummy substance was obtained in an amount of 1.17 g, which was found to be equal to the title compound on the basis of NMR analysis and thin layer chromatography.

Example XIII.

N-3- (N-cyclohexanecarbonyl-D-tryptophylthio) -2-D-methylpropanoyl-L-proline.

The procedure in Example 6 was repeated except that N-cyclohexanecarbonylglycine was replaced by 1.75 g of N-cyclohexanecarbonyl-D-tryptophan, 1.09 g of carbonyldiimidazole and 1.09 g of N- (3-mercapto-2- D-methylpropanoyl) -L-proline. Thus, a gum-like substance in an amount of 0.8 g was obtained and this substance was found to be equal to the title compound on the basis of the NMR analysis and thin layer chromatography.

Example XIV.

N- (3- (N-cyclohexanecarbonyl-D-phenylglycylthio) -2-D-methylpropanoyl] -L-proline.

The procedure in Example 6 was repeated except that N-cyclohexanecarbonylglycine was replaced by 1.30 g of N-cyclohexanecarbonyl-D-phenylglycine, 0.97 g of carbonyldiimidazole and 0.98 g of N- (3-mercapto-2- D-methylpropanoyl) -L-proline. Thus, 0.35 g of a gummy substance was obtained and was found to be equal to the title compound based on NMR analysis and thin layer chromatography.

8202472 -29- 22547 / Vk / ts

Example XV.

N-3- (N-cyclohexanecarbonyl-D-methionylthio) -2-D-methylpropanoyl-L-proline.

The procedure described in Example 6 was repeated except that 5 N-cyclohexanecarbonylglycine was replaced by 1.30 g of N-cyclohexanecarbonyl-D-methionine, 1.09 g of carbonyldiimidazole and 1.09 g of N- (3-mercapto-2- D-methylpropanoyl) -L-proline. Thus, 0.59 g of gummy substance was obtained and was found to be equal to the title compound based on NMR analysis and thin layer chromatography.

Example XVI.

N- {3- (N-cyclohexanecarbonyl-D-glutarainylthio) -2-D-raethylpropanoyl "| - L-proline.,

The procedure described in Example VI was repeated except that. N-cyclohexanecarbonylglycine, was replaced by 0.5 g N-cyclohexanecarbonyl-D-glutaraine, 0.41 g of carbonyldiimidazole and 0.41 g of N- (3-mercapto-2-D-methyl-propanoyl) -L-proline . Thus, a gummy substance in an amount of 0.3 g was obtained and was found to be equal to the title compound based on NMR analysis and thin layer chromatography.

Example XVII.

N-3- (N-adamantanecarbonyl-D-alanylthio) -2-D-methylpropanoyl-L-proline.

The procedure in Example 6 was repeated except that N-cyclohexanecarbonylglycine was replaced by 1.2 g of N-adamantancarbonyl-D-analine, 0.97 g of carbonyldidmidazole and 0.98 g of N- (3-mercapto-2-D- methylpro-panoyl) -L-proline. Thus, 1.02 g of gummy substance was obtained and was found to be equal to the title compound on the basis of NMR analysis and thin layer chromatography.

Example XVIII.

Dicydohexylamine salt of N - [3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methylpropanoyl -L-proline.

1) 2 * 11 g of 3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methyl-ethanecarboxylic acid was suspended in 1.3 l of dry dichloromethane. To the suspension, 117 ml of triethylamine was added dropwise at a temperature of -15 ° C with stirring, after which 104 ml of pivaloyl chloride was added dropwise at a temperature below -5 ° C with stirring and the mixture was further stirred for 30 minutes.

2) 101 g of L-proline was slurried in 1.2 l of dry dichloromethane - 8202472 *, - - ♦ ·· '* · t r -30- 22547 / Vk / ts. To the suspension, 112 ml of trimethylsilyl chloride was added dropwise at -15 ° C with stirring, then 122 ml of triethylamine was added dropwise at a temperature below -5 ° C with stirring and the mixture was further stirred at -15 ° C for 30 hours. minutes. To the resulting mixture, the solution prepared under 1) was added and the mixture was stirred with cooling for 15 minutes, then at ambient equipment for 1 hour. After completion of the reaction, 1 L of cold distilled water was added to the reaction mixture under cooling and then 90 ml of concentrated hydrochloric acid was added dropwise. The mixture was stirred for 10 minutes and the organic layer was separated, washed twice with a saturated NaCl solution and dried over magnesium murasulfate. To the filtrate was added 326 ml of dicyclohexylamine and 800 ml of acetonitrile and the mixture was added for 30 minutes stirred. Then 800 ml of acetonitrile was added four times at 10 minute intervals and the resulting mixture was stored overnight at a temperature of 5 ° C. The precipitate was filtered off under reduced pressure and dried in air to obtain a crude sample of the title compound. The crude sample was dissolved in 3 L of dichloromethane under an elevated temperature and then filtered. The filtrate was stirred at ambient temperature for 30 minutes and again 750 ml of acetonitrile was added four times at 10 minute intervals and the resulting mixture was stored at 5 ° C overnight. The precipitate was filtered and air-dried to obtain a colorless sample of the title compound in an amount of 40 µg. The melting point was 190-191 ° C and the D2 value = -24.5 (C = 1.0, MeOH).

Example XIX.

The calcium salt of N- (3- (N-cyclohexanecarbonyl-D-alanylthio) -2-D-methylpropanyl] -L-proline.

404 g of the dicyclohexylamine salt, prepared according to Example XVIII, were suspended in 2 liters of ethyl acetate and 2.5 L of 0.5 N KHSOjj was added to the suspension and the mixture was stirred for 10 minutes.

The organic layer was separated, washed with distilled water and dried over magnesium sulfate. The filtrate was evaporated under reduced pressure and the residue was dissolved in 1 * 2 1 acetone. 1.2 L of distilled water and 36.6 g of calcium carbonate were added to the acetone solution and the mixture was stirred vigorously for 1 hour at 40 ° C. After cooling, the mixture was filtered and the filtrate evaporated under reduced pressure.

8202472 -31-22547 / Vk / ts i

The resulting syrup was suspended in 1 liter of acetone and the solution was stirred for 5 hours. The precipitate was filtered off with suction, dried in air, then dried under reduced pressure, to obtain a white powder of the title compound in an amount of 212 g. Det ^ value = -47.2 ° (C = 1.0, MeOH).

In summary, it can be stated that according to the invention proline derivatives have been obtained corresponding to formula 1, stated on the forraule sheet, in which R is an acyl group attached to the pC-amino group of the amino acid and selected from the group consisting of cyclopropanecarbonyl, cyclo-10 hexanecarbonyl and adamantanocarbonyl groups, A is glycine, sarcosine or "I-D-amino acid residue and having an οί-carbonyl group forming a thiol ester with a sulfur atom, or a pharmaceutically acceptable salt thereof, and the invention further relates to a process for preparing this substance. The proline derivatives are suitable for the treatment of hypertension caused by angiotensin II and as a means for the treatment of cardiac insufficiency (inadequate functioning).

8202472

Claims (14)

A proline derivative, characterized in that it corresponds to formula 1, stated on the formula sheet, wherein R is an acyl group attached to the 5-amino group of the amino acid and selected from a group consisting of cyclopropanecarbonyl, cyclohexanoarbonyl and adamantane -earbonyl groups A is glycine, sarcosine or £-D-amino acid residue and with a (--carbonyl group which forms a thiol ester bond with a sulfur bomb, or a pharmaceutically acceptable salt thereof. A proline derivative according to claim 1, characterized in that A is an amino acid residue selected from a group consisting of glycine, sarcosine, D-alanine, D-leucine, D-methionine, D-glutamine, D-phenylalanine, D-trypto phane and D-phenylglycine. Proline derivative according to claim 1, characterized in that the 2-methylpropanoyl portion of the compound of formula 1 has a D-configuration. Proline derivative according to claim 3, characterized in that R is a cyclohexane carbonyl group and A is D-alanine. 5. Process for the preparation of a proline derivative, characterized in that a proline derivative is prepared according to formula 1, stated on the formula sheet, wherein R is an acyl group bonded to the amino group of the amino acid and is selected from a group consisting of cyclopropanecarbonyl, cyclohexanocarbonyl and adamantane carbonyl group, A is glycine, sarcosine or a D-amino acid residue and with a 1-carbonyl group forming a thiol ester bond with a sulfur atom, or a pharmaceutically acceptable salt thereof, by a compound of formula R-A -S-CH 2 CH (CH 2) COOH, where A 'is glycine, sarcosine, or a protected or unprotected <-D-amino acid residue, R has the same meaning as indicated above or a reactive derivative thereof in to react with a compound of formula 3, 30 indicated on the formula sheet, wherein R 'is a hydrogen atom or a carboxyl protecting group or a reactive derivative thereof in an inert organic solvent at a temperature between and -50 and 20 ° C, a possible protecting group is removed from the reaction product and optionally the reaction product is converted into a pharmaceutically acceptable salt. Process according to claim 5, characterized in that the reactive derivative of the compound of formula R-A'-S-CH 2 CH (CH 2) COOH is an activated amide, acid halide, activated ester or a mixed acid anhydride. 7. Process according to claim 5, characterized in that the: compound 8202472 -33-22547 / Vk / ts of formula R-A'-S-CH2CH (CH2) C00H is reacted with a compound of formula 3, listed on the formula sheet, or a reactive derivative thereof in the presence of a carbodiimide. 8. Process according to claim 5, characterized in that the reactive derivative of the compound of formula 3 has a silyl group which is introduced in the imino group of the compound. 9. Process according to claim 5, characterized in that the inert organic solvent used is tetrahydrofuran, dioxane, dimethylformamide, hexamethylphosphotriamide, chloroform, trichloromethane or acetontrile. A process for the preparation of a proline derivative, characterized in that a pDOline derivative of the formula 1, stated on the formula sheet, is prepared, wherein R is an acyl group bound to the ^-amino group of the amino acid and selected from the group consisting of from cyclopropanecarbonyl, cyclohexanecarbonyl and adamantancarbonyl groups, A is glycine, sarcosine or 15 cl-D-arainoic acid residue and with an od-carbonyl group forming a thiolester bond with a sulfur atom or a pharmaceutically acceptable salt thereof, by a compound of formula R-A ' -OH, where A 'is glycine, sarcosine or a protected or unprotected O-D-amino acid residue and R has the same meaning as indicated above, or to react a reactive derivative thereof with a compound of formula 4, mentioned on the formula sheet, wherein Rr is a hydrogen atom or a carboxyl protecting group, or a reactive derivative thereof in an inert organic solvent at a teraperature between -50 and 20 ° C, a possible protecting group is removed from the reaction product and the reaction product is optionally converted into a pharmaceutically acceptable salt. Web method according to claim 10, characterized in that the reactive derivative of the compound of formula R-A'-OH is an activated amide, acid halide, activated ester or a mixed acid anhydride. 12. Process according to claim 10, characterized in that the compound 30 of formula R-A-OH is reacted with a compound of formula 4 in the presence of a carbodiimide. Process according to claim 10, characterized in that the inert organic solvent is tetrahydrofuran, dioxane, dimethylformamide, hexamethylphosphotriamide, chloroform, dichloromethane or acetonitrile. 3514. A process for the manufacture of a medicament for combating hypertension and cardiac insufficiency, characterized in that a proline derivative of formula 1, wherein R and A have the same significance as stated in claim 1, is in a drug-suitable dosage form 8202472 '* ί · -'. · ·· 4 ~ Γ * -3 ^ - 22547 / Vk / ts brought. A method of preparing a proline derivative according to claim 1, characterized in that the preparation is carried out according to a method known for analogous compounds. 8202472