SK47695A3 - Boropeptide derivatives, method of their preparation and pharmaceutical agents containing these derivatives - Google Patents

Abstract

Boronic peptide derivs. of formula (I) are new. R = H, 1-4C alkyl, 1-4C alkoxy or 2-5C alkanoyl; R1 = Ph or cyclohexyl; R2 = an imidazolyl gp. of formula (a) or (b); R5 = H or -4C alkyl; R3, R4 = H; or R3+R4 = residue of a dihydroxy cpd.

Description

Technical field

The invention relates to boropeptide derivatives, to a process for their preparation and to their therapeutic use.

SUMMARY OF THE INVENTION

The compounds of the invention correspond to the general formula I

RH:

OR ³

OR ⁴ (I) in which

R represents a hydrogen atom or a straight or branched alkyl group having 1 to 4 carbon atoms or a straight or branched -CO-alkyl group in which the alkyl radical contains 1 to 4 carbon atoms, or a straight or branched -CO4-alkyl group in which the alkyl radical contains 1 to 4 carbon atoms,

R ¹ represents a phenyl group or a cyclohexyl group,

R ² represents a group R ⁵ R 6 or a group

NH 4 NH 2 = z

^R5 '\ wherein ^R is H or alkyl of 1 to 4 carbon atoms, and

R ³ and R ⁴ each represent a hydrogen atom or together represent a radical of a dihydroxylated compound such as 2,3-butanediol, 2,3-dimethyl-2,3-butanediol or (1a1a, 3a, 5a) -2,6,6 -trimethylbicyclo [3.1.1] -heptane-2,3-diol ((+) - alpha · pinanediol).

As defined by the general substituents R ³ and R ⁴ , the compounds of the invention have 3 or 7 asymmetric centers. The compounds of the invention may exist in the form of optical or geometric isomers in pure form or in the form of mixtures, and these forms are also within the scope of the invention. Preferred compounds of the invention having 3 asymmetric centers are D-alanyl-L-prolinamide derivatives having the (2 (R)) configuration, while preferred compounds of the invention having 7 asymmetric centers are D-alanyl-N- (derivatives) (4,6-methano-1,3,2-benzodioxaborol-2-yl) methyl] -L-proline amide having the (3aS), (2) R configuration, 3aalpha, 4beta, 6beta, 7aalpha //.

The compounds of the invention may exist in the form of the free bases or in the form of addition salts with pharmaceutically acceptable acids or bases, and these forms are also within the scope of the invention.

Preferred compounds of the invention are those wherein R ^a is a group

wherein ^R is H or alkyl of 1 to 4 carbon atoms. Among these compounds, selective compounds of the invention are those wherein R is hydrogen or straight or branched (C 1 -C 4) alkyl, R ¹ is phenyl, and R ³ and R ^{4 are} each hydrogen.

Finally, the most interesting compound of the invention is (R) - [1 - [(D-phenyl-alanyl-L-prolyl) amino] -4- (1H-imidazol-4-yl) butyl] boronic acid and its addition salts with pharmaceutically acceptable acids and bases.

Compounds of the invention wherein R ² is a group can be synthesized according to the following Reaction Scheme 1.

Reaction scheme 1

Borotripeptide of formula II (wherein R ^x is as defined above, R ³ and R ⁴ together represent the radical of a dihydroxy compound as defined above, R ⁶ represents either a hydrogen atom when R ⁷ represents a -CO-alkyl group in which the alkyl group the residue contains 1 to 4 carbon atoms in a straight or branched carbon chain, or -CO ₂ -alkyl group in which the alkyl residue contains 1 to 4 carbon atoms in a straight or branched carbon chain, when R ⁷ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms in a straight or branched carbon chain), is reacted with a compound of formula III (wherein ^R is H or alkyl of 1 to 4 carbon atoms) in a solvent such as dioxane, to afford of formula la. Then, if it is desired to obtain a compound of formula Ib (wherein R, R ¹ and R ^p are as defined above), a compound of formula Ia may be reacted with hydrochloric acid or a boron trichloride.

Compounds of the Invention in which ^RÄ represents a group;

NH wherein ^R is hydrogen or alkyl having 1 to 4 carbon atoms, may be synthesized according to Scheme second

Reaction scheme 2

(IV)

2) HCl

Ί

Reaction Scheme 2 (continued)

Reaction Scheme 2 (continued)

R ^ff R ⁷

OR '(Ic)

OR ^

NH

KCI

• NH //

HCl • 1 '- 3aS- (3aalpha, 4beta, 6beta, 7aalpha) -2- (3-bromopropyl) 3a, 5,5-trimethylhexadyhydro-4,6-methano-1,3,2-benzodioxaborol of formula IV reacted with sodium iodide in a solvent such as acetone to give [3aS- (3aalpha, 4beta, 6beta, 7aalpha) -2- (3-iodopropyl) -3a, 5,5-trimethylhexahydro-4,6-methano- l, 3,2-benzodioxaborole of formula V, which is condensed with a compound of formula VI (wherein ^R is as hereinbefore defined) to give a compound of formula VII, this reaction being carried out in a solvent such as tetrahydrofuran, at a temperature between -78 and 20 [deg.] C. Then, in accordance with a procedure analogous to that described by Mattheson in Organomettalics (1984), 3, 614, a compound of formula VII is reacted with dichloromethyllithium in the presence of zinc chloride in a solvent such as tetrahydrofuran , at a temperature between -100 and 20 ° C to give the compound of formula VIII, which is d o reaction with lithium bis (trimethylsilyl) amide, which reaction is carried out in a solvent such as tetrahydrofuran at a temperature between -78 and 20 ° C. This gives a compound which is reacted with hydrochloric acid in a solvent such as dioxane to give (R) -alpha-aminobutaneboronic acid hydrochloride of formula X, which is reacted with a diol of formula XI (wherein R ³ and R ³⁾ . ⁴ together are a radical as defined above dihydroxy compound) to form a hydrochloride salt of formula XII, then the compound of formula XII is condensed with an activated form of the dipeptide of the formula XIII (where ^R1 is as defined above, ^R represents a hydrogen atom when R ⁷ is CO -alkyl in which the alkyl radical contains 1 to 4 carbon atoms in a straight or branched carbon chain, or -CO 4 -alkyl in which the alkyl radical contains 1 to 4 carbon atoms in a straight or branched carbon chain when R ⁷ represents straight or branched (C 1 -C 4) alkyl, and X represents either 1-pyrrolidinyl-2,5-dione or 2-methylpropyl (carbonyl group) to form a compound which is reacted with hydrochloric acid to give a compound of formula Ic. Then, in order to obtain a compound of formula Id (in which R, R ¹ and R ⁵ are as defined above), the compound of formula Ic can be reacted with hydrochloric acid or boron trichloride.

The starting compounds used in the process of the invention are either commercially available or are described in the literature or can be prepared by the procedures described therein or by methods known to those skilled in the art.

The tripeptides of formula II are thus described in European patent application 0293881.

N-Acetyl-D-phenylalanyl-L-proline, 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha-N-acetyl-D-phenylalanyl-N- / 4-bromo-1- (3a) , 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl] -L-prolinamide and 1- (N -) (1,1-dimethylethoxy) -D-phenylalanyl] Proline is described by Kettner et al in J. Biol Chem (1990), 265, 18289.

[3aS- (3aalpha, 4beta, 6beta, 7aalpha)] - 2- (3-Bromopropyl) -3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol is described in European application 0293881.

2- [1,1-dimethylethyl] dimethylsilyl] -N, N-dimethyl-1H-imidazole-1-sulfonamide is described by Ngochindo in J. Chem. Bathroom. Perkin Trans. (1990), 1, 1645.

The preparation of the compounds of formula XII is described in French patent application FR 9404287.

The preparation of compounds of formula XIII (wherein R ⁶ represents a straight or branched -CO 4 -alkyl group in which the alkyl radical contains 1 to 4 carbon atoms and R ⁷ represents a straight or branched alkyl group containing 1 to 4 carbon atoms) is carried out using the corresponding starting compounds of formula XIII (wherein R ⁷ is hydrogen) according to the procedure of Bajusz in J. Med. Chem., (1990), 33, 1729-1735.

In the following, the invention will be further elucidated by means of specific examples thereof, the examples being illustrative only and not in any way limiting the scope of the invention, which is clearly defined by the definition of the claims. The chemical structures of the obtained compounds were confirmed by elemental microanalysis and infrared and nuclear magnetic resonance spectroscopy. The numbers of the compounds given in the headings of the examples correspond to those given in the table below. The ratio in brackets refers to the acid / base molar ratio.

DETAILED DESCRIPTION OF THE INVENTION

Example 1 (compound 1) / 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha) - N-Acetyl-D-phenylalanyl-N - [4- (1H-imidazol-1-yl) - 1- (3a, 5,5-trimethylhexahydro4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl] -L-proline amide hydrochloride (1: 1).

1.1. (3aS-) 2 (R), 3aalpha, 4beta, 6beta, 7aalpha) - N-Acetyl-D-phenylalanyl-N - [4- (ΙΗ-imidazol-1-yl) -1- (3a, 5) 5-trimethylhexahyro-4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl] -L-prolinamide g (4.8 mmol) / 3aS- / 2 (R), 3aalpha, 4beta, 6beta .

7aalfa // - N-acetyl-D-phenylalanyl-N- / 4-bromo-l- (3, 5,5-trimethylhexahydro-4,6-methano-l, 3,2-benzodioxaborol-2-yl) butyl / 1-L-proline amide and 0.902 g (9.6 mmol) of 1H-imidazole are dissolved in 10 ml of dioxane. The solution was heated at 80 ° C for 6 hours, then the solvent was evaporated in vacuo. The residue is taken up in 50 ml of dichloromethane and the organic phase is then washed successively with 5% aqueous sodium bicarbonate solution and saturated sodium chloride solution. This phase is then dried over sodium sulphate, filtered and evaporated. The residue is purified by chromatography on a column of silica gel, eluting with a methanol / dichloromethane (9:95; v / v) mixture. 1.2 g of product are obtained in the form of an oil.

Yield: 41%, [.alpha.] D @ ²⁰ = -77.5 DEG (c = 1.5, chloroform).

D

1.2. (3aS-) 2 (R), 3aalpha, 4beta, 6beta, 7aalpha) - N-Acetyl-D-phenylalanyl-N - [4- (1H-imidazol-1-yl) -1- (3a, 5) 5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl-L-proline amide hydrochloride (1: 1)

Dissolve 1.2 g (1.97 mmol) of 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha] - N-acetyl-D-phenylalanyl-N- / 4- (3-chloro) in 3 ml of chloroform. ΙΗ-Imidazol-1-yl) -1- (3α, 5,5-trimethylhexahyro-4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl] -L-proline amide and the resulting solution was cooled to 0 C. To the solution is added 20 ml of a 0.1N solution of hydrochloric acid in isopropanol, and the mixture is stirred for 15 minutes at 0 ° C, then concentrated in vacuo. The residue is dried over phosphorus pentoxide and triturated in ether. 1 g of product is obtained in the form of a solid amorphous substance.

MP: 99 ^e C, yield: 78%, / alpha / ²⁰ = -90.4 ° (c = 1.1, chloroform).

Example 2 (compound 2) (R) -N-Acetyl-D-phenylalanyl-N- [1-borono-4- (1H) imidazol-1-yl) butyl] -L-prollnamide hydrochloride (1: 1)

Dissolve 700 mg (1,2 mmol) of (3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha) - N-acetyl-Dphenylalanyl-N- / 4- (ΙΗ-imidazole) in 6 ml of anhydrous dichloromethane. -1-yl) -1- (3α, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl] -L-proline amide 13 hydrochloride. The solution was cooled to -78 ° C and 5 mL (5 mmol) of a 1M solution of boron trichloride in dichloromethane was added dropwise over 15 minutes. The mixture is stirred for 15 minutes at -78 ° C and for 45 minutes at 0 ° C, then 15 ml of water are slowly added. The solution is stirred for 30 minutes at 0 ° C, then 10% acetic acid solution is added. The phases are separated and the aqueous phase is extracted three times with 10 ml of ether, then evaporated to dryness. The residue is purified by chromatography on a Biogel P2 column, eluting with a 10% acetic acid solution. The product-containing fraction was concentrated and the residue was triturated in ether. The product is obtained in the form of a white powder which is diluted with 2 ml of water. A solution is obtained which is purified by chromatography on a Bíograd AG 1W8 column (OH cycle), eluting with water and 1N hydrochloric acid solution. The product-containing fraction was concentrated and the residue was triturated in ether.

50 mg of product are obtained.

Melting point: 168-175 ° C (dec.), Yield: 10%, [.alpha.] D @ ²⁰ = -118.5 DEG (c = 0.6, water).

D

Example 3 (compound 3) / 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha) - N-Acetyl-D-phenylaanyl-N- / 4- (1H-imidazol-4 (5) -yl) ) -1- (3a, 5,5-trimethylhexahydro4,6-methano-1,3,2-benzodoxaborol-2-yl) butyl] -L-proline amide hydrochloride (1: 1)

3.1 / 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha] - alpha (3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) - 1H-imidazole-4 (5) -butanamine hydrochloride (2: 1)

3.1.1. (3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha) -2- (3-iodopropyl) -3a, 5,5-trimethylhexahydro-4,6-methano14

1,3,2-benzodioxaborole

A solution of 37 g (122 mmol) of [3aS- (3aalpha, 4beta, 6beta, 7aalpha)] - 2- (3-bromopropyl) -3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol and 72 Sodium iodide (7 g, 488 mmol) in acetone (500 ml) was heated at reflux for 24 hours. The solvent is then evaporated and the residue is taken up in a mixture of 500 ml of ether and 100 ml of water containing 1 g of sodium sulfite. The organic phase is dried over magnesium sulphate, then filtered and evaporated. 40 g of product are obtained, which product is used immediately in the next step. Yield: 95%.

3.1.2. [3aS- (3aalpha, 4beta, 6beta, 7aalpha) -2 - [(1,1-dimethylethyl) dimethylsilyl] -N, N-dimethyl-4 (5) - / 3- (3a, 5,5-trimethylhexahydrone-4,6) -methano-1,3,2-benzodioxaborol-2-yl) propyl] -1H-imidazole-1-sulfonamide

70.5 g (244 mmol) of 2 - [(1,1-dimethylethyl) dimethylsilyl] -N, N-dimethyl-1H-imidazole-1-sulfonamide are dissolved in 250 ml of tetrahydrofuran. The reaction mixture was cooled to -78 ° C and 152 mL (244 mmol) of a 1.6 M solution of n-butyllithium in hexane was added. The mixture is stirred for one hour at -78 ° C, then 40 g (115 mmol) of < 3 > S- (3aalpha, 4beta, 6beta, 7aalpha) -2- (3-iodopropyl) -3a, 5,5- Trimethylhexahydro-4,6-methano-1,3,2-benzodoxaborol in solution in 100 ml of tetrahydrofuran The reaction mixture is stirred for one hour at -78 and 20 ° C and then for 2 hours at 20 ° C. The reaction mixture is then poured into 350 ml of an ice / water mixture containing 14.5 g (121 mmol) of sodium hydrogen sulfate, the aqueous phase is extracted three times with 100 ml of ether, then the ether phases are combined, dried over magnesium sulphate, filtered and evaporated. The residue is purified by chromatography on a column of silica gel, eluting with a 20% ethyl acetate / hexane solution.

45 g of product are obtained.

Yield: 73%, [.alpha.] D @ ²⁰ +12.5 DEG (c = 1.9, chloroform).

D

3.1.3. [3aS- / 2 (S), 3aalpha, 4beta, 6beta, 7aalpha] -4-Chloro-4- (3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2- yl) butyl / 2 - / (1,1-dímetylety1) dimethylsilyl / N, N-dimethyl-lH-imidazol-l-sulfonic acid amide

A solution of 8.3 g (98 mmol) of dichloromethane in 100 mL of tetrahydrofuran was cooled to -100 ° C. To this cooled solution was added 39.1 mL (98 mmol) of a 2.5 M solution of n-butyllithium in hexane. The mixture is left at this temperature for 15 minutes, then 45 g (89 mmol) of [3aS- (3aalpha, 4beta, 6beta, 7aalpha)] - 2 - / (1,1-dimethylethyl) dimethylsilyl] -N, N-dimethyl are added. -4 (5) - [3- (3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) propyl] -1H-imidazole-1-sulfonamide in solution in 50 ml of tetrahydrofuran . The reaction mixture is left at -100 ° C for 15 minutes, then 9.8 g (70 mmol) of a solution of zinc chloride in 50 ml of tetrahydrofuran is added. The mixture was allowed to warm to 20 ° C over 16 hours. It is evaporated under vacuum and the residue is taken up in a mixture of 200 ml of dichloromethane and 50 ml of water. The phases are separated and the aqueous phase is extracted with 100 ml of dichloromethane. The organic phases are combined, dried over magnesium sulphate, filtered and evaporated. The color residue is purified by chromatography on silica gel, eluting with a mixture of ethyl acetate and hexane (20:80; v / v).

40 g of product are obtained in the form of a colorless oil.

Yield: 80%, [.alpha.] D @ ²⁰ +15.9 DEG (c = 2.65, chloroform).

D

3.1.4. / 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha // -4 (5) - / 4 amino-4- (3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2benzodioxaborol- 2-yl) butyl N, N-dimethyl-1H-imidazole 1-sulfonamide hydrochloride (1: 1)

12.6 g (78 mmol) of 1,1,1,3,3,3-hexamethylsilazane are dissolved in 80 ml of tetrahydrofuran and 31 ml (78 mmol) of a 2.5 M solution of n-butyllithium in hexane are added. The mixture is left for one hour at-78 ^and C, followed by the addition of 40 g (71 mmol) of / 3a / 2 (R), 3aalfa, 4beta, 6?, 7aalfa // - 4 (5) - / 4-chloro-4- (3, 5,5-trimethylhexahydro-4,6-methano-l, 3,2-benzodioxaborol-2-yl) butyl / 2 - / (1,1-dimethylethyl) dimethylsilyl / N Of N-dimethyl-1H-imidazole-1-sulfonamide in solution in 80 mL of tetrahydrofuran. The mixture was stirred for 1 hour at -78 ° C and then for 16 hours at 20 ° C. The reaction mixture was then cooled to -78 ° C, 78 mL (312 mmol) of a 4N solution of hydrochloric acid in dioxane was added thereto, and the mixture was stirred at -78 ° C for one hour and at room temperature for 2 hours. temperature 20 ° C. The mixture was evaporated in vacuo and the residue taken up in 200 ml of chloroform. The mixture was evaporated. 32 g of product are obtained in the form of an oil which is triturated in ether.

The product is obtained as a solid.

Yield: 89%, mp: 90-92 ° C, [α] ²⁰ D +11 ° (c = 1, methanol).

D

3.1.5. (3aS-) 2 (R), 3aalpha, 4beta, 6beta, 7aalpha-alpha (3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodoxaborol-2-yl) -1H- imidazole-4 (5) -butanamine hydrochloride (2: 1) g (70 mmol) / 3aS- / 2 (R), 3aalpha, 4beta, 6beta,

7aalfa // - 4 (5) - / 4-amino-4- (3, 5,5-trimethylhexahydro-4,6-metano1,3,2-benzodioxaborol-2-yl) butyl / N, N-dimethyl- 1 H-Imidazole-1-sulfonamide hydrochloride in solution in 200 ml of 4N hydrochloric acid is heated to boiling for 3 hours. The solution is extracted four times with 100 ml of ether, then evaporated to dryness. The residue is taken up in 100 ml of methanol and 11.9 g (70 mmol) of (1R- (1alpha, 2alpha, 3alpha, 5alpha) -2,6,6-trimethylbicyclo [3.1.1] heptane-2,3-) are added to the obtained fraction. The mixture was stirred for 16 hours at 20 ° C and then evaporated to dryness to give 27 g of product as an oil which was triturated in ether to give the product as a solid.

Melting point: 75-80 ° C.

2.3 N-acetyl-D-phenylalanyl-L-proline ester with 1-hydroxypyrrolidine-2,5-dione g (20 mmol) of N-acetyl-D-phenylalanyl-L-proline is suspended in a mixture of 100 ml of ethyl acetate and 5 ml of dimethylformamide. To the obtained suspension was added 2.53 g (22 mmol) of 1-hydroxypyrrolidine-2,5-dione and the resulting mixture was cooled to 0 ° C. 4.53 g (22 mmol) of 1,3-dicyclohexylcarbodiimide in solid form are then added in small portions. The mixture was stirred at 20 ° C for 20 hours, then the resulting suspension was filtered. The filtrate is washed successively with 20 ml of 5% sodium bicarbonate solution and saturated sodium chloride solution, then dried over magnesium sulfate. The residue is triturated in ether.

8 g of a glassy product are obtained, which product is used as is in the following stage.

3.3 [3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha] - N-acetyl

D-phenylalanine-N- / 4- (lH-imidazol-4 (5) -yl-1- (3, 5,5trimetylhexahydro-4,6-methano-l, 3,2-benzodioxaborol-2yl) butyl / -L -prolinamide hydrochloride (1: 1)

2.4 g (6.4 mmol) / 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha-alpha- (3a, 5,5-trimethylhexahydro-4,6-methano-1,3), 2-Benzodioxaborol-2-yl) -1H-imidazole-4 (S) -butanamine hydrochloride was dissolved in 20 mL of dichloromethane and 2.5 g (6.4 mmol) of N-acetyl-D- ester was added. of phenylalanyl-L-proline with 1-hydroxypyrrolidone-2,5-dione, then the resulting mixture is cooled to -30 ° C. Triethylamine (3.6 mL, 25.6 mol) was added dropwise and the mixture was stirred at -30 ° C for 2 hours and at 20 ° C for 2 hours. 20 ml of a 5% aqueous sodium hydrogen carbonate solution are then added and the aqueous phase is extracted twice with 20 l of dichloromethane. The organic phases are combined, dried over sodium sulfate, filtered and evaporated.

The residue is taken up in 10 ml of isopropanol, then 64 ml of a 0.1N solution of hydrochloric acid in isopropanol are added at 0 ° C. After evaporation, the residue is decolorized with charcoal in ethyl acetate and purified by chromatography on a Sephadex LH-20 column eluting with methanol. The product-containing eluate was evaporated and the residue triturated in ether. 2 g of product are obtained in the form of a colorless solid.

Melting point: 130-135 ° C, yield: 51%, [.alpha.] D @ ²⁰ = -112.1 DEG (c = 1, chloroform).

D

Example 4 (compound 4) (R) -N-Acetyl-D-phenylalanyl-N- [1-borono-4- (1H-imidazol-4 (5) -ylbutyl) -L-proline amide hydrochloride (1: 1) g (3 mmol) / 3aS- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha - N-acetyl-D-phenylalanine-N- / 4- (1H-imidazol-4 (5) -yl-1) (3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl-L-proline amide hydrochloride was dissolved in 30 ml of dichloromethane and cooled to -78 °. 12 ml of a 1M solution of boron trichloride in dichloromethane is then added dropwise to the cooled solution, and the reaction mixture is left at -78 ° C for 15 minutes, then placed on ice and stirred at 0 ° for 45 minutes. C. 30 ml of water are added, then the phases are separated, the organic phase is extracted twice with 25 ml of 10% acetic acid solution and the aqueous phase is extracted three times with 25 ml of ether, the aqueous phases are combined, concentrated and diluted with 10 ml of methanol and evaporated. The residue was then purified by bio-gel P2 column chromatography using 10% acetic acid solution as eluent followed by column chromatography on Bio-gel P2 using 1 mM hydrochloric acid as eluent.

800 mg of product are obtained.

Melting point: 155-160 ° C, yield: 52%, [.alpha.] D @ ²⁰ -98.5 DEG (c = 0.65, water).

D

Example 5 (compound 5) / 3aS- / 2 / R / S (R) //, 3aalpha, 4beta, 6beta, 7aalpha // - / 2- / 2 - /// 4- (1H-Imidazol-4-yl) ) -1- (3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) butyl / amino / carbonyl / pyrrolidin-1-yl] 2-oxo -1- ( phenylmethyl) ethyl / carbamate hydrochloride (1: 1)

To 7.2 g (7.2 mmol) of N - [(1,1-dimethylethoxy) carbonyl] D-phenylalanyl-L-proline in solution in 30 ml of tetrahydrofuran is added 2.3 ml (21 mmol) of N at ambient temperature. methylmorpholine. The reaction mixture was cooled to -20 ° C and then 2.72 mL (21 mmol) of isobutyl formate was added. The reaction mixture was stirred for 15 min at-20 ^and C, followed by the addition of 7.5 g (20 mmol) / 3a / 2 (R), 3aalfa, 4beta, 6?, 7aalfa // - a- ( 3a, 5,5-trimethylhexahydro-4,6-methano-1,3,2-benzodioxaborol-2-yl) -1H-imidazol-4 (5) -butanamine in solution in 10 ml of chloroform and 5.6 ml (40 ml). mmol) triethylamine. The reaction mixture is stirred for 30 minutes at -20 ° C and then for 24 hours at ambient temperature. The mixture was diluted with 300 mL of ethyl acetate and washed successively with 200 mL of water and 200 mL of saturated sodium chloride solution, then dried over sodium sulfate. The solvent is evaporated to dryness and the residue is taken up in 50 ml of dichloromethane. The hydrochloride is prepared by adding 20 ml of a solution of isopropanol in 0.1N hydrochloric acid, then purified by column chromatography on Lichoprep RP 18, eluting with a gradient of acetonitrile in 0.02N hydrochloric acid (20% to 100%).

5 g of product are obtained in the form of the hydrochloride.

Melting point: 115-120 ° C, yield: 45%, [.alpha.] D @ ²⁰ -106 DEG (c = 1, chloroform).

Example 6 (Compound 6) (R) - 1 - [(D-Phenylalanyl-L-prolyl) amino] -4- (1H-imidazol-4-yl) butyl] boronic acid hydrochloride (3: 1)

From 698 mg (1 mmol) 1,1-dimethylethyl- (3aS-) / 2 / R / S (R) //, 3aalpha, 4beta, 6beta, 7aalpha // - / 2- / 2 - /// 4- ( H-imidazol-4-yl) -1- (3, 5,5-trimethylhexahydro-4,6-methano-l, 3,2-benzodioxaborol-2-y1) -butyl / amino / carbonyl / pyrrolidine-l- yl] -2-oxo-1- (phenylmethyl) ethyl] carbamate hydrochloride was prepared as in Example 4 and purified on a Lichoprep RP 18 column eluting with acetonitrile in 0.02 N hydrochloric acid to give 300 mg of the title compound.

Melting point: 194 DEG C., [.alpha.] D @ ²⁰ -131 DEG (c = 1.3, water).

D

Example 7 (compound 7)

1,1-Dimethylethyl- [3aS-] 2 / R / S (R) //, 3aalpha, 4beta, 6beta, 7aalpha // - / 2- / 2 - /// 4- / 5-methyl-1H-imidazole 4-yl) -1- (3, 5,5-trimethylhexahydro-4,6-methano-l, 3,2-benzodioxaborol-2-yl) butyl / amino / carbonyl / pyrrolidin-l-yl / -2 -oxo-1- (phenylmethyl) ethyl / carbamate hydrochloride (1: 1)

This product was prepared according to the procedure described in Example 5 of (3a, S- / 2 (R), 3aalpha, 4beta, 6beta, 7aalpha) -5-methyl-alpha- (3a, 5,5-trimethylhexahydro-4,6). methano-l, 3,2benzodioxaborol-2-yl) -lH-imidazole-4-butanamine hydrochloride. 600 mg of product are obtained.

Melting point: 85-90 ° C, [α] ²⁰ D -53.2 ° (c = 0.86, methanol). about

The following table shows the chemical structures and chemical properties of some of the compounds of the invention. In the R ³ column, the -C ₆ H _s symbol represents a phenyl group and the symbol

-C ₆ H _xi represents a cyclohexyl group. In the Salt column, the HCl symbol represents the hydrochloride and the brackets indicate the acid / base molar ratio. In the column Tt (° C), the symbol d indicates that melting takes place on decomposition.

table

Table (continued)

g about J 130-135 155-160 115-120 194 85-90 0 rh Ό (0 and V> tn a M about E U 0 ι-H V_i '0 m N <H 0 i-4 about rh (0 T m ú «. what cn in 1 VD about -106 (1; chloroform) and <0 Ό ABOUT rH> n r-3 · * » 1 n H rh rH 0 q (0 CN e n ^Β lf) 1 co co * o rh 0 0] f-j about ·· X rt <-H 1 H G ·· X H fH rH u ·· CS Ή f — 1H G ·· X n fH G ·· X H <· · CS <of .alpha. X 1 m. X 1 ΛΔ m cx x 1 Y-_ _υ x 1 \ 4-p cs. CS <\ J 2 · ^ x / 2 ^ _ / <\ j 3: / 2___ / <\ J 2 --- cC about ts' X C 1 Wk x Ό G 1 wn x Ό G 1 wn x J = 1 x G 1 x —1 x G ABOUT G 1 x G ABOUT G l m «n x G G ABOUT ABOUT G 1 x 1 ÍR m x G G ABOUT ABOUT G 1 > G n and VO r *

Table (continued)

The compounds of the invention were tested to determine their effect against thrombin and trypsin in vitro using the following assays.

1) Precipitation of human fibrinogen by bovine thrombin

The test compound or vehicle (10 µl) is incubated for 2 minutes at 37 ° C in human fibrinogen solution (200 µl, 2 mg / ml in saline). Then 200 µl of bovine thrombin dissolved in distilled water is added. The final thrombin concentration is 0.5 units of NIH / ml. The mixture is agitated and the time in seconds required to form a visible fibrin network is recorded. Inhibition of fibrin formation is quantified by calculating the concentration of compound that increases the time required to precipitate the fibrin network by 100% (CA).

2) Precipitation of human fibrinogen by human thrombin

The test compound or vehicle (10 µl) is incubated for 2 minutes at 37 ° C in a human fibrinogen solution (200 µl, 2 mg / ml in saline). Then 200 µl of human thrombin dissolved in distilled water is added. The final thrombin concentration is 2 units of NIH / ml. The mixture is agitated and the time in seconds required to form a visible fibrin network is recorded. Inhibition of fibrin formation is quantified by calculating the concentration of compound that extends the time required to precipitate the fibrin network by

100% (CA).

'xoo ¹

3) Coagulation of rat plasma with bovine thrombin

Rat male CDs weighing 150-200 g are anesthetized with nembutal (60 mg / kg, 0.1 ml / kg). Anesthetized rats are bled from the retro-orbital canal for 3.8% trisodium citric acid salt. Plasma is prepared from blood by centrifugation at 3600 g and ambient temperature for 15 minutes. The test compound or vehicle (10 µl) is incubated with 200 µl of said plasma at 37 ° C for 2 minutes, then 200 µl of bovine thrombin solution is added. The final thrombin concentration is 0.75 units of NIH / ml. The time in seconds to coagulate rat plasma is recorded. Inhibition of thrombin is quantified by calculating the concentration that increases the time required to coagulate plasma by 100%.

4) Aggregation of rabbit platelets by human thrombin

The animals were bled by cardiac puncture to a 3.8% trisodium citric acid salt (1 volume per 9 volumes of blood). The collected blood is centrifuged at 250 g for 10 minutes. The platelet-rich plasma (P ₃ P) thus obtained is isolated and the platelet count is determined. To the P ₃ P plates, 2 ng / ml prostacyclin in solution in ice-cold Tris buffer (pH = 9.0) is added. The mixture is centrifuged at 110 g for 10 minutes, then decanted. Prostacyclin in a solution (50 mM, pH 12) of sodium hydroxide is added again to a final concentration of 200 ng / ml. Plates P1P are centrifuged again at 800 g for 10 minutes. Platelet-poor plasma is removed and the remaining pellet is suspended in a volume of tyrod containing 200 ng / ml prostacyclin equal to a starting volume of P ₃ P. This suspension is centrifuged at 800 g for 10 minutes. The suspension of the sediment pellet and centrifugation of the suspension are repeated once again under the same conditions. The final pellet is suspended in a solution of prosthodylin-free tyrodine, then the suspension is allowed to stand for 2 hours to allow complete prostacyclin elimination. Aggregation of these platelets is induced by human thrombin at a final concentration of 0.3 NIH / ml. Changes in optical density are recorded using a 4 channel aggregometer. Test compound is added to the plate suspension (maximum volume added 3 µl) 2 minutes before thrombin addition. The concentration which inhibits aggregation of the said 50% (C _a).

5) Effect on bovine trypsin

The test compound or its vehicle (50 .mu.l) is incubated for 5 minutes at room temperature with 50 .mu.l _of bovine trypsin dissolved in tris-HCl buffer (pH 8.0). The final thrombin concentration is 229 units / ml. The reaction is initiated by the addition of a substrate which is N-alpha-benzoyl-L-arginine-4-nitroaniline (50 µM). The mixture is incubated for 20 minutes at ambient temperature, then the optical density of the released is measured

4-nitroaniline at 405 nm. The concentration of 4-nitroaniline is calculated from the calibration curve by subtracting the optical density corresponding to the blank (100 µl buffer + 50 µl substrate). The concentration which inhibits 50% of the enzyme activity of trypsin (C _a).

The compounds of the invention were tested in parallel to inhibit rat plasma coagulation ex vivo. The test compound is administered intravenously, orally or subcutaneously to the test compound prior to blood collection. The time required for coagulation of plasma by thrombin is then measured as in Test 3).

The compounds of the invention are thrombin inhibitors and have CA and Cl in the range of between 10 ^-a and 10 ^-6 M in this regard. Nexoo exhibits no or little bovine trypsin inhibitory activity, demonstrating their specificity.

These compounds inhibit rat plasma coagulation at doses less than 1 mg / kg by intravenous administration, and are equally effective for oral and subcutaneous administration.

The compounds of the invention may be useful in all clinical indications associated with thrombosis, or in clinical indications in which three shoe complications could occur.

For this purpose, the compounds of the invention may be formulated in forms suitable for oral, parenteral or intravenous administration such as tablets, dragees, gelatin capsules, capsules, drinkable or injectable suspensions or solutions and the like, in combination with suitable excipients and in amounts which it allows the patient a daily dose of the active compound of 1 to 1000 mg in a single dose or in several divided doses.

Claims (10)

PATENT CLAIMS Compounds of general formula I in which: R represents either a hydrogen atom or a straight or branched alkyl group having 1 to 4 carbon atoms or a straight or branched -CO-alkyl group in which the alkyl radical contains 1 to 4 carbon atoms or a straight or branched -CO4-alkyl group in wherein the alkyl radical contains 1 to 4 carbon atoms, R ^x represents either a phenyl or cyclohexyl, R ² is either wherein ^R is H or alkyl of 1 to 4 carbon atoms, and R ³ and R ⁴ each represent a hydrogen atom or together represent the radical of the dihydroxy compound, in the form of optical or geometric isomers, in the pure state or in the form of mixtures as well as addition salts thereof with pharmaceutically acceptable acids and bases. Compounds according to claim 1, wherein the residue of the dihydroxylated compound is selected from the group consisting of 2,3-butanediol, 2,3-dimethyl-2,3-butanediol, and (1alpha, 3alpha, 5alpha) 2,6,6-trimethylbicyclo (3.1.1) heptane-2,3-diol / (+) - alpha-pinanediol /. Compounds according to claims 1 and 2 having, according to the meanings of R ³ and R ^{4, the} configuration (2 (R)) or (3aS), (2), 3aalpha, 4beta, 6beta, 7aalpha //. Compounds according to any one of claims 1 to 3, in which R ² represents a group in which R ⁵ represents a hydrogen atom or an alkyl group containing 1 to 4 carbon atoms. Compounds according to claim 4 of the general formula I, in which R represents a hydrogen atom or a straight or branched alkyl group having 1 to 4 carbon atoms, R ¹ represents a phenyl group and R ³ and R ⁴ each represent a hydrogen atom. 6. A compound according to claim 1 which is (R) - [1- (D-phenylalanyl-L-prolyl) amino] -4- (1H-imidazole-4-butyl) boronic acid as well as its addition salts with pharmaceutically acceptable salts. acids and bases. 7. A process for the preparation of compounds of claim 1 of formula I, wherein R ² is a group where ^R is H or alkyl having 1 to 4 carbon atoms, characterized in that the borotripeptid formula II Br in which R ¹ is as defined in claim 1, R ³ and R ⁴ together represent the radical of the dihydroxy compound as defined in claim 1, R ^e represents either a hydrogen atom when R ⁷ represents a straight or branched -CO-alkyl group, wherein the alkyl radical contains 1 to 4 carbon atoms, or a straight or branched -CO 4 -alkyl group, wherein the alkyl radical contains 1 to 4 carbon atoms, when R ⁷ is a hydrogen atom or a straight or branched alkyl group containing 1 to 4 atoms % of carbon, is reacted with a compound of formula (III) (CIII) wherein R ⁵ is as defined in claim 1, to form a compound of formula Ia and then, when formula 1b is to be obtained, Q K HCl in which R, R ^x and R ^a are as defined in claim 1, reacting a compound of formula Ia with hydrochloric acid or a boron trichloride. A process for the preparation of compounds according to claim 1, wherein R ² is a group NH wherein ^R is H or alkyl of 1 to 4 carbon atoms, characterized in that the hydrochloride salt of formula XII (XII) wherein R ³ and R ⁴ together are a radical of dihydroxy compound as defined in Claim l and R ^p is as referred to in claim 1, condenses with an activated form of a dipeptide of formula xiii * wherein R ¹ is as defined in claim 1, R ^e represents either a hydrogen atom when R ⁷ represents a straight or branched -CO-alkyl group in which the alkyl moiety contains 1 to 4 carbon atoms, or a straight or branched -CO4-alkyl group in which the alkyl radical contains 1 to 4 carbon atoms, when R ⁷ represents a hydrogen atom or a straight or branched alkyl group containing 1 to 4 carbon atoms and X represents either 1-pyrrolidinyl-2,5-dione or 2-methylpropyloxycarbonyl, to form a compound which is reacted with hydrochloric acid to form for a compound of formula 1c 0R ^J < OR ⁴ HCl (c) then in the case where the compounds of formula Id wherein R, R ¹ and R ^p are as defined in claim 1, reacting a compound of formula Ic with a hydrochloric acid or a boron trichloride. Medicament, characterized in that it comprises a compound according to any one of claims 1 to 5. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5 in combination with an appropriate pharmaceutical excipient.