SI9200269A - Amino acid derivates - Google Patents

Abstract

Amino acid derivatives of the formula: Sl 9200269 A in which R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aralkanoyl, heterocyclylcarbonyl, or a group with the formula: R®HN R 2 represents alkyl, cycloalkyl or aralkyl; R3 represents hydrogen and R4 represent hydroxy, or R3 and R4 together represent oxo; R5 represents alkoxycarbonyl or alkylcarbamoyl; R6 and R7 together represent trimethylene or tetramethylene, optionally substituted by alkyl or adjacent carbon atoms with tetramethylene; R8 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aroyl, aralkanoyl, or heterocyclylcarbonyl; and R9 represents alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, cyanoalkyl, carbamoylalkyl, alkylthioalkyl, alkoxyalkyl or alkoxycarbonylalkyl, and pharmaceutically acceptable acid the addition salts of those compounds of formula I which are basic, inhibit aspartyl proteases of viral origin and to be given in the form of medicines for prophylaxis or treatment viral infections. It is generally known to prepare them methods.

Description

R ² represents alkyl, cycloalkyl or aralkyl; R ³ represents hydrogen and R ⁴ represents hydroxy, or R ³ and R ⁴ together represent oxo; R ⁵ represents alkoxycarbonyl or alkylcarbamoyl; R ⁶ and R ⁷ together represent trimethylene or tetramethylene, optionally substituted with alkyl or on adjacent carbon atoms with tetramethylene; R ⁸ represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aroyl, aralkanoyl, or heterocyclylcarbonyl; and R ⁹ represents alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, cyanoalkyl, carbamoylalkyl, alkylthioalkyl, alkoxyalkyl or alkoxycarbonylalkyl, and the pharmaceutically acceptable acid addition salts of those compounds of formula I that are basic, inhibit viral source aspartyl proteases and are used medicines for the prophylaxis or treatment of viral infections. They can be prepared by commonly known methods.

F.HOFFMANN-LA ROCHE AG

DERIVATES AMINO ACID 15

The present invention relates to amino acid derivatives. It relates in more detail to amino acid derivatives of the general formula

FPHN

in which R4 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, 30 aralkanoyl, heterocyclylcarbonyl, or a group of formula

R®HN (s);

R ⁹

R 2 represents alkyl, cycloalkylalkyl or aralkyl; R ⁴ represents hydrogen and R ⁴ represents hydroxy; or R ^ and R ⁴ together represent an oxo; R4 represents alkoxycarbonyl or alkylcarbamoyl; R ^ and R? together represent trimethylene or tetramethylene, optionally substituted by alkyl or adjacent carbon atoms with tetramethylene; R4 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aroyl, aralkanoyl, or heterocyclylcarbonyl; and R4 represents alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, cyanoalkyl, carbamoylalkyl, alkylthioalkyl, alkoxyalkyl or alkoxycarbonylalkyl, and to the pharmaceutically acceptable acid addition salts of those compounds of formula I which are basic.

The compounds and salts mentioned above are novel and have valuable pharmacological properties. In particular, they inhibit aspartyl proteases of viral origin and can be used in the prophylaxis and treatment of viral infections, in particular infections caused by HIV and other retro viruses.

The subject of the present invention are the compounds of formula I and the abovementioned salts alone and for use as therapeutically effective substances, the process for the preparation of said compounds and salts, the intermediates used in that process, the medicaments containing the said compounds and salts, the use of said compounds and salts in the prevention or control of diseases, in particular in the prophylaxis or treatment of viral infections, and the use of said compounds and salts for the preparation of medicaments for the prophylaxis or treatment of viral infections.

As used herein, the term alkyl, alone or in combination, means a straight or branched alkyl group containing not more than eight, preferably not more than four, carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec.butyl, tert.butyl, n-pentyl, n-hexyl and the like. The term alkoxy, alone or in combination, means an alkyl ether group in which the term alkyl has the foregoing meaning, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec.butoxy, tert-butoxy and the like. The term cycloalkyl, alone or in combination, means a cycloalkyl group containing 3 to 8, preferably 3 to 6, carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. The term aralkh means an alkyl group as previously defined in which one hydrogen atom is replaced by an aryl group, e.g. a phenyl or naphthyl group, optionally bearing one or more substituents selected from alkyl, alkoxy, halogen, trifluoromethyl, hydroxy, nitro, amino and the like. Examples of aralkyl groups are benzyl, 4-chlorobenzyl, 4-methoxybenzyl, 2-phenylethyl, 2-naphthylethyl and the like. The term aralkyloxycarbonyl means a group of the formula aralkyl-O-C (O) - in which the term aralkyl has the aforementioned meaning. The term alkanoyl means an acyl group derived from an alkanecarboxylic acid such as acetyl, propionyl, butyryl, valeryl,

4-methylvaleryl and the like. The term aroyl means a benzoyl or naphthoyl group, optionally bearing one or more substituents selected from alkyl, alkoxyl, halogen, trifluoromethyl, hydroxy, nitro, amino and the like, such as benzoyl, p-chlorobenzoyl, 3,5-dichlorobenzoyl, 1-naphthoyl and the like. The term aralkanoyl means an acyl group derived from aryl-substituted alkanecarboxylic acid such as phenylacetyl, 3-phenylpropionyl (hydrocinamoyl), 4-phenylbutyryl, (2-naphthyl) acetyl, 4-chloro-,

4-amino- or 4-methoxyhydro-cinnamoyl and the like. The term heterocyclylcarbonyl means a group of the formula -CO-Het in which Het is a saturated, partially unsaturated or aromatic, monocyclic, bicyclic or tricyclic heterocycle containing one or more hetero atoms selected from nitrogen, oxygen and sulfur, as applicable substituted on one or more carbon atoms by halogen, alkyl, alkoxy, oxo, etc. and / or on a secondary nitrogen atom (i.e.-NH-) with alkyl, aralkoxycarbonyl, alkanoyl, phenyl or phenylalkyl, or on a tertiary nitrogen atom (i.e. = N-) with an oxide, which is attached via a carbon atom. Examples of such Het groups are pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, pyrrolyl, imidazolyl, irazolyl, pyridyl, pyrazinyl, pyrimidinyl, furyl, thienyl, triazolyl, oxazolyl, thiazolyl, indolyl, quinolyl, isoquinolinyl, 4,3-quinolinyl. -tetrahydroisoquinolyl, quinoxalinyl, beta-carbolinilin-like. The term halogen means fluorine, chlorine, bromine and iodine.

Pharmaceutically acceptable acid addition salts are formed between the base compounds of formula I and inorganic acids, e.g. hydrohalic acids such as hydrochloric acid and hydrobromic acid, sulfuric, solitic and phosphoric acids, etc., or organic acids, e.g. acetic acid, citric acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid and p-toluenesulfonic acid, etc.

The compounds of formula I contain at least three asymmetric carbon atoms and are therefore present in the form of optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisemic racemates or mixtures and diastereoisomic racemates.

The present invention encompasses all these forms.

A particular group of compounds of formula I comprises those in which it represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aralkanoyl or heterocyclylcarbonyl.

In the compounds of formula I, R 1 is preferably alkoxycarbonyl, in particular tert-butoxycarbonyl, or a group of formula (i), in which R 1 represents araloxycarbonyl, in particular benzyloxycarbonyl, aroyl, in particular 3,5-dichlorobenzoyl, or heterocyclylcarbonyl, in particular 2-quinolylcarbonyl, and R4 represents alkyl, especially isopropyl or tert-butyl, aralkyl, especially benzyl, cyanoalkyl, especially cyanomethyl, carbamoylalkyl, especially carbamoylmethyl, alkylthioalkyl, especially methylthiomethyl, or alkoxycarbonylalkyl, especially methoxycarbonylmethyl. R 2 preferably represents aralkyl, especially benzyl. Preferably R 4 is hydrogen and R 4 represents hydroxy. If R ^ represents alkoxycarbonyl, it is preferably methoxycarbonyl and if R ^ represents alkylcarbamoyl, it is preferably tert-butylcarbamoyl. R ^ and R ^ together are preferably unsubstituted tetramethylene.

It is apparent from the above that particularly preferred compounds of formula I are those in which R1 represents tert-butoxycarbonyl or a group of formula (i) in which R1 represents benzyloxycarbonyl, 3,5-dichlorobenzoyl or 2-quinolylcarbonyl and R4 represents isopropyl , tert-butyl, benzyl, cyanomethyl, carbamoylmethyl, methylthiomethyl or methoxycarbonylmethyl, R ^ represents benzyl, R ^ represents hydrogen and R ^ represents hydroxy, R ^ represents methoxycarbonyl or tert-butylcarbamoyl and R ^ and RT together represent unsubstituted tetramethylene.

A particularly preferred compound of formula I is:

2 (S) - [3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane -carboxamide.

Other particularly preferred compounds of formula I are:

2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide,

2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1 (R) -cyclohexane-carboxamide ,

2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-phenylalanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide ,

2 (S) - [3 (S) - [[N- (2-quinolylcarbonyl) -L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1 (R) -cyclohexane -carboxamide,

2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -S-methyl-L-cysteinyl] annno] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide,

2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-valyl] amino] -2-oxo-4-phenylbutyl] -Nert-t-butyl-1 (R) -cyclohexane-carboxamide, ² ( S) - [3 (S) - [[N- (benzyloxycarbonyl) -0-methyl-L-aspartyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) - cyclohexane-carboxamide,

2 (S) - [3 (S) - [[N- (3,5-dichlorobenzoyl) -L-asparagmyl] amino] -2- (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R ) -cyclohexane-carboxamide,

2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -3-methyl-L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamidine methyl 2 (S) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -1 (R) cyclohexane-carboxylate.

Examples of other interesting compounds of formula I are:

2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide ,

2 (S) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexanecarboxamide,

2 (R) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexanecarboxamide and

2 (R) - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide.

In accordance with the method provided by the present invention, compounds of formula I and pharmaceutically acceptable acid addition salts of those compounds which are basic are prepared such that (a) for the preparation of a compound of formula I in which R4 represents alkoxycarbonyl or aralkoxycarbonyl , R 4 represents hydrogen and R 4 represents hydroxy, the compound of the general formula is treated

wherein R ^ ^a represents alkoxycarbonyl or aralkoxycarbonyl and R ² , R 5, r 6 and R ^{7 have} the meanings given above, with an acid, or (b) by reaction of a compound of the general formula

in which R ² , R 4, R 4, r ⁵ , r 6 j _n r 7 p _re j have the indicated meaning, with an acylating agent which introduces the group Rf as previously defined, or (c) for the preparation of a compound of formula I in which Rf represents a group of formula (i) by reaction of a compound of general formula IV

in which R ² , R ⁴ , r ⁴ r ⁵ , r 6, r ⁷ j _n r 9 p _re j have the above meaning, with an acylating agent which introduces the group R 1 as defined above, or (d) for the preparation of a compound of formula I, in which R4 and R4 together represent oxo, by oxidation of a compound of formula I, in which R4 represents hydrogen and R4 represents hydroxy, and / or (e) optionally by separating a mixture of diastereoisomeric racemates into diastereoisomeric racemates or optically pure diastereoisomers , and / or (f) optionally separating the mixture of diastereoisomers into optically pure diastereoisomers, and / or (g) optionally converting the resulting basic compound of formula I into a pharmaceutically acceptable acid addition salt.

Treatment of a compound of formula II with an acid according to embodiment (a) of the process yields a compound of formula I wherein Rf represents alkoxycarbonyl or aralkoxycarbonyl, R4 represents hydrogen and R4 represents hydroxy. The type of acid used in this embodiment depends essentially on the nature of the substituent R ^ ³ 'present in the starting material of formula II. If R ^ ^a represents alkoxycarbonyl, e.g. tert.butoxycarbonyl, the treatment is preferably carried out using a strong organic acid, in particular an organic sulfonic acid such as alkanesulfonic acid, e.g. methanesulfonic acid, or aromatic sulfonic acid, e.g. benzenesulfonic acid, p-toluenesulfonic acid, mesitylensulfonic acid, etc. and in the presence of an organic solvent which is inert under the reaction conditions, e.g. alkanols such as methanol, ethanol, etc. However, halogenated alkanecarboxylic acid, e.g. trifluoroacetic acid, etc., instead of organic sulfonic acid. Where R ^a represents an aralkoxycarbonyl, for example. benzyloxycarbonyl, the treatment is preferably carried out using a solution of halogen, e.g. hydrogen chloride, in alkanol, e.g. methanol, and in the presence of an organic solvent which is inert under the reaction conditions, e.g. halogenated aliphatic hydrocarbon such as dichloromethane, etc.

The reaction of a compound of formula III with an acylating agent according to embodiment (b) of the process can be carried out in a manner known per se using the appropriate acid or its reactive derivative as an acylating agent. Suitable reactive derivatives are acid halides, e.g. acid chlorides, acid anhydrides, mixed anhydrides, activated esters, etc. If the acylime agent is such that it introduces a group of formula (i), the reaction is expeditiously carried out using an acid in the presence of a condensing agent, e.g. dicyclohexylcarbodiimide or benzotriazol-1yloxy-tris (dimethylamino) phosphonium hexafluorophosphate, and in the presence of a base such as triethylamine, ethyldiisopropylamine and the like. The reaction is conveniently carried out at a temperature between about 0 ° C and about room temperature, preferably at room temperature.

The reaction of a compound of formula IV with an acylating agent according to embodiment (c) of the process can be carried out in a manner known per se using the appropriate acid or its reactive derivative as an acylating agent. Suitable reactive derivatives are acid halides, e.g. acid chlorides, acid anhydrides, mixed anhydrides, activated esters, etc. The reaction is conveniently carried out at a temperature between about 0 ° C and about room temperature, preferably at room temperature.

Oxidation according to embodiment (d) of the process can be carried out in a manner known per se for the oxidation of secondary alcohols into ketones. So e.g. oxidation can be carried out using pyridinium dichromate in dimethylformamide, pyridinium chlorochromate in dichloromethane, sulfur trioxide / pyridine complex in dimethylsulfoxide, oxalyl chloride and triethylamine in dimethylsulfoxetrolyethoxylate trifluoromethoxybluoxytetrahydrofluoroimethoxidine

Optional separations according to embodiments (e) and (f) of the process can be performed by conventional methods, e.g. by column chromatography, thin layer chromatography, high pressure liquid chromatography, etc.

The conversion of a basic compound of formula I into a pharmaceutically acceptable salt according to the embodiment (g) of the process can be carried out by treatment in a conventional manner with an inorganic acid, e.g. hydrochloric acid such as hydrochloric or hydrobromic acid, sulfuric, solitic and phosphoric acid, etc., or with an organic acid, e.g. acetic acid, citric acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid and ptoluenesulfonic acid, etc.

The compounds of formula II which are used as starting materials in carrying out (a) of the process are novel and form a further object of the present invention. Prepare them according to the following reaction scheme in which R ^ ³ , R2, r6 and R? the meaning given above.

FP * HN

Reaction scheme

(Π)

In view of the above Reaction Scheme, a compound of formula V is reacted in the first step with a compound of formula VI to give a compound of formula VIL The reaction is carried out under normal Grignard reaction conditions; e.g. in an organic solvent which is inert under reaction conditions such as ether, e.g. diethyl ether, and at a temperature between about 0 ° C and about 40 ° C, preferably at about room temperature.

In the next step, the compound of formula VII is converted to the compound of formula VIII by reaction with 2,2-dimethoxypropane in the presence of a strong organic acid, preferably an organic sulfonic acid, such as p-toluenesulfonic acid. The reaction is conveniently carried out at about room temperature.

Subsequently, the compound of formula VIII is oxidized to the compound of formula IX. The oxidation is preferably carried out using alkali metal permanganate as potassium permanganate at about room temperature. The oxidation is advantageously carried out in a solvent system comprising a mixture of water, alkanecarboxylic acid as a freezing inert organic solvent which is immiscible with them, e.g. aromatic hydrocarbons such as benzene, toluene, etc., and in the presence of a phase transfer catalyst.

Finally, the compound of formula IX is converted to the compound of formula II by esterification or amidation. The esterification or amidation is carried out by reaction of a compound of formula IX with a suitable alkanol or amine by methods known per se. .

The compounds of formulas V and VI used to prepare the compounds of formula II are known compounds or analogs of known compounds which can be prepared in a similar manner to known compounds. In addition, the Examples below are detailed information pertaining to the preparation of certain compounds of Formula VI. The compounds of formulas VII, VIII and IX, on the other hand, are novel and are the subject of the present invention.

The compounds of formula III used as starting materials in carrying out (b) of the process are novel and also form the object of the present invention. They can be prepared by cleaving an alkoxycarbonyl or aralkoxy carbonyl group from a compound of formula I wherein R1 represents an alkoxycarbonyl or aralkoxycarbonyl group. The cleavage can be performed by methods known per se. So e.g. in the case where R1 represents alkoxycarbonyl, cleavage can be carried out using a strong inorganic acid as hydrochloric acid, or a strong organic acid, e.g. trifluoroacetic acid, preferably at about 0 ° C to about room temperature, but if R1 represents an aralkoxycarbonyl group, cleavage can be carried out using hydrogen in the presence of a noble metal catalyst, e.g. palladium on charcoal, in an organic solvent that is inert under reaction conditions, e.g. in alkanol as ethanol, etc., or in the alkanecarboxylic acid ester as ethyl acetate, and preferably at about room temperature.

The compounds of formula IV used as starting materials in carrying out (c) the process are novel and form a further object of the present invention. They can be prepared by cleaving an alkoxycarbonyl or aralkoxycarbonyl group from a compound of formula I, in which R1 represents a group of formula (i) and R1 represents an alkoxycarbonyl or aralkoxycarbonyl group. The cleavage can be performed by methods known per se. So e.g. in the case where R < 1 > represents an alkoxycarbonyl group, cleavage can be carried out using a strong inorganic acid as hydrochloric acid, or a strong organic acid, e.g. trifluoroacetic acid, preferably at about 0 [deg.] C. to about room temperature, but if r8 represents an aralkoxycarbonyl group, cleavage can be carried out using hydrogen in the presence of a noble metal catalyst, e.g. palladium on carbon, in an organic solvent which is inert under reaction conditions, e.g. in alkanol as ethanol, etc., or in the alkanecarboxylic acid ester as ethyl acetate, and preferably at about room temperature.

As previously mentioned, the compounds of formula I and the pharmaceutically acceptable acid addition salts of those compounds which are basic, inhibit aspartyl proteases of viral origin and are useful in the treatment and prophylaxis of viral infections, in particular infections caused by HIV and other retro viruses.

In vitro inhibition of HIV protease by the compounds of the present invention can be demonstrated by the following assay:

HTV protease was expressed in E. coli and partially purified from soluble bacterial extracts by fractionation with ammonium sulfate (0-30%). Protease activity was determined using the protected heptapeptide of succinyl-Val-Ser-Gln-Asn-Phe-Pro-Ile isobutylamide as a substrate. The cleavage of the substrate was quantified by measuring the production of H-Pro-Ile isobutylamide by spectrophotometric evidence on the Infinite Proline.

The 1.25 mM substrate was dissolved in 125 mM citrate buffer (pH 5.5) containing 0.125 mg / ml Tween 20. A solution (10 μΥ) of various concentrations of the test compound (dissolved in methanol or dimethylsulfoxide and diluted with water containing 0.1% Tween 20) and 10 μΐ protease were added to 80 μ \ of the upper buffered substrate. Digestion was performed at 37 ° C for a fixed period of time and ended with the addition of 1 ml of color reagent [30 µg / ml isatin and

1.5 mg / ml 2- (4-chlorobenzoyl) benzoic acid in 10% acetone in ethanol (vol./vol.)]. The solution was heated in a water bath and then the pigmented residues were redissolved in 1 ml of 1% pyrogallol in 33% water in acetone (w / v / v). The optical density of the solution was measured spectrophotometrically at 599 nm. The formation of H-Pro-Ile isobutylamide in the presence of test compound was compared with controls and determine the concentration of test compound required to give 50% inhibition (I50) ^by means of the curve plotted from the various concentrations of the test compound uprabljene.

The in vitro antiviral activity of the compounds of formula I can be demonstrated by the experiment described below:

this experiment uses HTLV-III (RF strain) grown in C8166 cells (human CD4 +

T lymphoblastoid line) using RPM1 1640 medium with bicarbonate buffer, antibiotics and 10% fetal bovine serum.

Cell suspension was infected with tenfold TCID5q virus and allowed adsorption to progress for 90 minutes at 37 ° C. The cells were washed three times with medium. The assay was performed in 6 ml tissue culture tubes containing 2 x 10 ^ infected cells in 1.5 ml medium. The test compounds were dissolved, either in aqueous medium or in dimethylsulfoxide, in accordance with the solubility, and 15 μl of ratopine test compounds were added. The cultures were incubated at 37 ° C for 72 hours in a humidified atmosphere containing 5% carbon dioxide in the air. The cultures were then centrifuged and an aliquot of the supernatant (supernatant) solubilized with Nonidet P40 and subjected to antigen-based assays using a primary antiserum with specific reactivity to the viral p24 antigen and horseradish peroxidase detection system. The color formation is measured spectrophotometrically and plotted against the concentration of the test compound. Determine the concentration (I50) which gives 50% protection.

The results obtained in the above tests using representatives of the compounds of formula I are summarized in the following table:

Table

Compound HIV protease inhibition Ϊ5θ (nmol) Anti-HIV activity 150 (nmol) A 2 46 B 57 1000 C 1.4 4 D 1.3 NT E 1.9 NT F 2.0 NT

NT = not tested.

Compound A = 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] -amino] -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R ) -cyclohexanecarboxamide.

Compound B = 2 (R) - [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] -amino] -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R ) -cyclohexanecarboxamide.

Compound C = 2 (S) - [3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] -amino] -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide.

Compound D = 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -S-methyl-L-cysteinyl] -amino] -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl -l (R) -cyclohexanecarboxamide.

Compound E = 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -0-methyl-L-aspartyl] -amino] -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl -l (R) -cyclohexanecarboxamide.

Compound F = 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] -amino] -2 (R) hydroxy-4-phenylbutyl] -N-tert-butyl -l (R) -cyclohexanecarboxamide.

The compounds of formula I and the pharmaceutically acceptable salts of those compounds of formula I which are basic can be used as medicaments, e.g. in the form of pharmaceutical preparations. The pharmaceutical preparations may be administered enterally as oral, e.g. in the form of tablets, coated tablets, dragees, hard and soft gelatin capsules, solutions, emulsions or suspensions, nasally, e.g. in the form of nasal sprays (sprays), rectally, e.g. in the form of suppositories, or parenterally, as intramuscularly or intravenously, e.g. in the form of injection solutions.

For the preparation of pharmaceutical preparations, the aforementioned compounds and salts may be treated with therapeutically inert, inorganic or organic additives. Lactose, corn starch or derivatives thereof, talc, stearic acid or its salts may be used e.g. as such additives for tablets, coated tablets, dragees and hard gelatin capsules. Suitable additives for soft gelatin capsules are e.g. vegetable oils, waxes, fats, semi-solids and liquid polyols and the like. Depending on the nature of the effective ingredient, they are generally not required in the case of soft gelatin capsules. Suitable additives for making solutions and syrups are e.g. water, polyols, sucrose, invert sugar, glucose and the like. Suitable additives for the manufacture of injection solutions are e.g. water, alcohols, polyols, glycerin, vegetable oils and the like. Natural and hardened oils, waxes, fats, semi-liquid polyols and the like are suitable additives for the manufacture of suppositories.

Pharmaceutical preparations may also contain preservatives, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavors, osmotic pressure adjustment salts, buffers, coating agents or antioxidants. They may also contain other therapeutically effective substances.

Drugs containing a compound of formula I or a pharmaceutically acceptable salt of a basic compound of formula I and a therapeutically inert additive, as well as a process for making such drugs, are also objects of the present invention. Such a process involves the conversion of a compound of Formula I or its salts mentioned above into a galenic delivery form, together with a therapeutically inert additive, and optionally with one or more other therapeutically effective substances.

As mentioned previously, the compounds of formula I and their salts mentioned above may be used in the control or prevention of disease, especially in the prophylaxis or treatment of viral infections, in particular retroviral infections. Dosage can vary within wide limits and will, of course, be tailored to individual needs in each specific case. Generally, in the case of oral administration, a daily dosage of about 3 mg to about 3 g, preferably about 10 mg to about 1 g, is acceptable, although the upper limit may be exceeded if deemed appropriate. The daily dosage can be given as a single dose or in divided doses.

The following examples illustrate the present invention:

Example 1

A solution of 100 mg (0.20 mmol) 2 (S) - [[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamidine

3.3 mg (0.017 mmol) of p-toluenesulfonic acid in 3 ml of methanol was maintained at room temperature for 40 hours. The solvent was removed by evaporation and the residue was partitioned between 10 ml of dichloromethane and 2 ml of saturated sodium hydrogen carbonate solution. The organic solution was dried over anhydrous magnesium sulfate and evaporated to give 90 mg of rubber. The crude product was chromatographed on a silica gel column using 33% ethyl acetate / hexane for elution to give 45 mg of 2 (S) - [3 (S) - (tert-butoxyformamido) -2- (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide; MS m / e 447 [M + H] ⁺ .

2 (S) - [[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl-tert-butyl-1 (R) -cyclohexanecarboxamide used as the starting material was prepared as follows:

(i) a solution of 4.375 g (31 mmol) of trans-4-cyclohexen-1 (R), 2 (R) -dimethanol and 4.66 g (31 mmol) of tert-butyl dimethylsilyl chloride in 16 ml of dimethylformamide were stirred under nitrogen and cooled 5.30 g of imidazole was added and the mixture was allowed to warm to room temperature and then stirred overnight. 200 ml of water were added and the mixture was extracted with three 100 ml portions of diethyl ether. The combined extracts were washed with 100 ml of water, dried over anhydrous magnesium sulfate and evaporated to give 6.50 g of oil. This was chromatographed on a silica gel column using 10% ethyl acetate in hexane for elution to give 2.80 g of trans-1 (R), 2 (R) -bis [[(tert-butyl) - (dimethyl) silyloxy] methyl] - 4-cyclohexene as a colorless oil. Further elution of the column gave 2.576 g of trans-6 (R) - [[(tert-butyl) (dimethyl) silyloxy] methyl] -3-cyclohexene-1 (R) -methanol as a colorless oil;

MS m / e 257 [M + H] +.

(ii) A solution of 2.57 g (10 mmol) of the second product of paragraph (i) in 50 ml of ethanol was hydrogenated over 130 mg of 10% palladium-on-charcoal catalyst at room temperature and atmospheric pressure for 1 hour. The catalyst was removed by filtration and the filtrate was evaporated to give 2.30 g of trans-2 (R) - [[(tert-butyl) (dimethyl) silyloxy] methyl] -1 (R) -cyclohexanmethanol as a colorless oil;

MS m / e 259 [M + H] +.

(iii) A solution of 1.17 g (9.2 mmol) of oxalyl chloride in 23 ml of dichloromethane was stirred under nitrogen and cooled to -78 ° C. A solution was added over 6 minutes.

1.3 ml (1.43 g, 18 mmol) of anhydrous dimethyl sulfoxide in 5 ml of dichloromethane. The mixture was stirred for a further 2 minutes and then a solution of 2.30 g (8.9 mmol) of the product from paragraph (ii) in 10 ml of dichloromethane was added over a 9 minute period. The mixture was stirred for 20 minutes at -70 ° C and then 5.5 ml (3.99 g, 39 mmol) of triethylamine were added over a 6 minute period. The mixture was then allowed to warm to room temperature and 40 ml of water were added. The phases were separated and the aqueous solution extracted with two 50 ml portions of dichloromethane. The combined organic solutions were dried over anhydrous magnesium sulfate and evaporated to give 2.45 g of a colorless oil. This was purified by silica gel column chromatography using 2.5% ethyl acetate in hexane for elution to give 1.53 g of trans-2 (R) - [[(tert-butyl) (dimethyl) silyloxy] methyl] -1 (R) -cyclohexanecarboxaldehyde as a colorless gum; MS m / e 257 [M + H] +.

(iv) 3.46 ml (8.6 mmol) of 2.5M solution of n-butyllithium in hexane was added over 5 minutes to a stirred suspension of 3.8 g (8.06 mmol) of methyl triphenylphosphonium bromide in 23 ml of diethyl ether. The mixture was stirred at room temperature for 85 minutes and then a solution of 1.53 g (6.0 mmol) of the product of paragraph (iii) in 10 ml of diethyl ether was added over 5 minutes. The mixture was stirred for 140 minutes at room temperature, 20 ml of water was added and the phase was separated. The aqueous solution was extracted with two 20 ml portions of diethyl ether and the combined organic solutions were washed with 20 ml water, dried over anhydrous magnesium sulfate and evaporated to give 1.85 g of oil. This was purified by chromatography on a silica gel column using 1% ethyl acetate in hexane for elution to give 0.870 g of trans-1 (R) - [[(tert-butyl) - (dimethyl) silyloxy] methyl] 2 (S) - vinylcyclohexane as a colorless oil;

MS m / e 239 [M - CH ₃ ] +.

(v) A solution of 870 mg (3.4 mmol) of the product of paragraph (iv) in 6 ml of an IM solution of tetrabutylammonium fluoride in tetrahydrofuran was maintained at room temperature for 3.5 hours. The solvent was then evaporated and the residue taken up in 30 ml of water and extracted with three 20 ml portions of diethyl ether. The combined extracts were dried over anhydrous magnesium sulfate and evaporated to give 910 mg of a colorless oil. This was chromatographed on a silica gel column using 20% ethyl acetate in hexane for elution to give 435 mg of trans-2 (S) -vinyl-1 (R) -cyclohexanmethanol as a colorless oil;

MS m / e 141 [M + H] +.

(vi) A solution of 435 mg (3.1 mmol) of the product of paragraph (v) in 10 ml of pyridine was stirred under nitrogen and cooled in an ice bath. 265 gl (391 mg,

3.4 mmol) of methanesulfonyl chloride, the ice bath was removed and the mixture was stirred at room temperature for 4 hours. The solvent was removed by evaporation and the residue was partitioned between 30 ml of 2M hydrochloric acid and 30 ml of diethyl ether. The aqueous solution was extracted with two 30 ml portions of diethyl ether and the combined extracts washed with 30 ml of 2M hydrochloric acid, 30 ml of saturated sodium hydrogen carbonate solution and 30 ml of water, dried over anhydrous magnesium sulfate and evaporated to give 615 mg l ( R) - [(methanesulfonyloxy) methyl] -2 (S) 10 vinylcyclohexane as a colorless oil; MS m / e 219 [M + H] ⁺ .

(vii) A mixture of 1.55 g (7.1 mmol) of the product of paragraph (vi) and 935 mg (10.7 mmol) of lithium bromide in 20 ml of dimethylformamide was stirred for 50 hours at 60 ° C under nitrogen. The mixture was then poured into 250 ml of water and extracted with three 100 ml portions of diethyl ether. The combined extracts were washed with 200 ml of water, dried over anhydrous magnesium sulfate and evaporated to give 1.312 g of trans-l (R) -bromomethyl2 (S) -vinylcyclohexane as a colorless liquid; MS m / e 123 [M - Br] +.

(viii) A mixture of 172 mg (7.1 mmol) of magnesium chips and iodine crystals in 2 ml of tetrahydrofuran was stirred under argon and 1.113 g (5.5 mmol) of the product of paragraph (vii) was added over three minutes. The mixture was stirred and refluxed for 1 hour, then cooled to room temperature. A solution of 683 mg (2.74 mmol) of tert-butyl (La-formylphenethyl) carbamate in 5 ml of tetrahydrofuran was added over 10 minutes and the mixture was stirred for a further 2.5 hours at room temperature. Then 30 ml of 10% ammonium chloride solution was added, the mixture was adjusted to pH 2 with 2M hydrochloric acid and then extracted with three 25 ml portions of ethyl acetate. The combined extracts were washed with 25 ml of saturated sodium bicarbonate solution and 25 ml of water, dried over anhydrous magnesium sulfate and evaporated to give 1.19 g of oil. This was chromatographed on a silica gel column using 20% ethyl acetate / hexane for elution to give 630 mg of mixture 1 (S) [3 (S) - (tert-butoxyformamido) -2 (S) -hydroxy-4- phenylbutyl] -2 (S) -vinylcyclohexane and 1 (S) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -2 (S) vinylcyclohexane as a white crystalline solid; MS m / e 374 [M + H] ⁺ . The diastereomeric mixture was used in the next step without further purification.

(ix) A solution of 630 mg (1.7 mmol) of the product of paragraph (viii) in 20 ml of dimethylformamide was stirred and cooled to 0 ° C. 4.48 g (12 mmol) of pyridinium dichromate was added and the mixture was stirred at 0 ° C for 10 minutes. The cooling bath was removed and the mixture was stirred at room temperature for 6 hours, then poured into 170 ml of water. The resulting mixture was extracted with three 80 ml portions of ethyl acetate and the combined extracts washed with 100 ml water, dried over anhydrous magnesium sulfate and evaporated to give 640 mg l (S) - [3 (S) - (tert-butoxyformamido) - Zoxo-4-phenylbutyl] -2 (S) -vinylcyclohexane as a colorless oil which crystallizes on condition;

This product was used in the next step without further purification.

(x) A solution of 640 mg (1.7 mmol) of the product of paragraph (ix) in 35 ml of ethanol was stirred at 0 ° C and 368 mg (9.7 mmol) of sodium borohydride was added. The mixture was stirred at 0 ° C for 2.5 hours and then the solvent was removed by evaporation. The residue was partitioned between 100 ml of water and 100 ml of ethyl acetate, the phases separated and the aqueous phase extracted with separate 100 ml and 50 ml portions of ethyl acetate. The combined extracts were dried over anhydrous magnesium sulfate and resisted to give 595 mg of a white oily solid. This was chromatographed on a silica gel column using 20% ethyl acetate / hexane for elution to give 85 mg of 1 (S) - [3 (S) (tert-butoxyformamido) -2 (S) -hydroxy-4-phenylbutyl ] -2 (S) -vinylcyclohexane as a white waxy solid; MS m / e 374 [M + H] +. Further elution of the column gave 263 mg of l (S) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -2 (S) vinylcyclohexane as a white solid; MS m / e 374 [M + H] ⁺ .

(xi) A solution of 370 mg (0.99 mmol) of the second product of paragraph (x) and 225 mg (1.2 mmol) of p-toluenesulfonic acid monohydrate in 7 ml of 2,2-dimethoxypropane was maintained overnight at room temperature. The solution was diluted with 45 ml of diethyl ether and washed with two 40 ml portions of sodium hydrogen carbonate, dried over anhydrous magnesium sulfate and evaporated to give 530 mg of yellow oil. This was chromatographed on a silica gel column using 5% ethyl acetate / hexane for elution to give 230 mg of l (S) - [[4 (S) -benzyl-3 (tert-butoxycarbonyl) -2,2-dimethyl]. -5 (R) -oxazolidinyl] methyl] -2 (S) -vinylcyclohexane as a pale yellow oil; MS m / e 414 [M + H] ⁺ .

(xii) A solution of 670 mg (4.2 mmol) of potassium permanganate in 7 ml of water was added to a mixture of 230 mg (0.56 mmol) of the product of paragraph (xi), 123 mg of Aliquat 336 and 0.65 ml of glacial in 8 ml of benzene. The mixture was stirred vigorously overnight at room temperature and then 428 mg of sodium metabisulphite was added. 24 ml of diethyl ether and 2.5 ml of 2M citric acid were added, the mixture was stirred for 10 minutes and then filtered.

The phases were separated and the aqueous phase extracted with two 10 ml portions of diethyl ether. The combined extracts were washed twice with 10 ml of water each, then dried over anhydrous magnesium sulfate and evaporated to give 340 mg of oil. This was chromatographed on a silica gel column using 5% methanol / dichloromethane for elution to give 210 mg of 2 (S) - [[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5 (R) ) -oxazolidinyl] methyl] -1 (R) -cyclohexane carboxylic acid as a colorless gum, which was used in the next step without further purification.

(xiii) Mixture of 210 mg (0.49 mmol) of the product referred to in paragraph (xii) 67 mg (0.49 mmol)

1-Hydroxybenzotriazole hydrate, 101 mg (0.49 mmol) of dicyclohexylcarbodiimide and 52 μϊ (36 mg, 0.49 mmol) of tert.butyl amine in 2 ml of dimethylformamide were stirred for 20 hours at room temperature under nitrogen. The mixture was filtered and the solid was washed with 2 ml ethyl acetate. The combined filtrates were evaporated and the residue partitioned between 10 ml of ethyl acetate and 10 ml of saturated sodium hydrogen carbonate solution. The phases were separated and the aqueous phase extracted with two 10 ml portions of ethyl acetate. The combined extracts were washed with 10 ml of saturated sodium chloride solution, dried over anhydrous magnesium sulfate and evaporated to give 280 mg of an oily solid. This was extracted with two 1 ml portions of diethyl ether and discarded. The ether solutions were evaporated to give 260 mg of crude product. This was chromatographed on a silica gel column using 22% ethyl acetate / hexane for elution to give 109 mg of 2 (S) - [[4 (S) -benzyl-3 (tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl] -N-tert-butyl-1 (R) cyclohexanecarboxamide as a white solid; MS m / e 487 [M + E < ^{+ &gt};] ⁺ .

Example 2

A mixture of 35 mg (0.1 mmol) 2 (S) - [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl (R) -cyclohexanecarboxamide and 44 mg (0.1 mmol) N -benzyloxycarbonyl-Lasparagine pentafluorophenyl ester in 1 ml of dioxane was stirred for 16 hours at room temperature under nitrogen. The solvent was evaporated, the residue taken up in 10 ml of ethyl acetate and the solution was washed with two 3 ml portions of 2M hydrochloric acid, 3 ml 2M sodium hydroxide solution and 3 ml saturated sodium chloride solution, dried over anhydrous magnesium sulfate and evaporated. The residue was chromatographed on a silica gel column using 6% methanol / dichloromethane for elution to give 34 mg of a white solid. This was recrystallized from dichloromethane / hexane to give 14 mg of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) 20

L-asparaginyl] -amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexanecarboxamide as a white solid from m.p. 195-199 ° C;

MS m / e 595 [M + H] < + >.

2 (S) - [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexanecarboxamide was used as starting material as follows:

mg (0.1 mmol) 2 (S) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] N-tert-butyl-1 (R) -cyclohexanecarboxamide [obtained as described in the first paragraph of Example 1], was dissolved in 2 ml of ethyl acetate and the solution cooled to 0 ° C. 0.2 ml of saturated hydrogen chloride solution in ethyl acetate was added and the mixture was allowed to stand overnight at room temperature. A further 1.0 ml of hydrogen chloride in ethyl acetate was then added, the mixture was stirred for a further 4 hours at room temperature and then evaporated to dryness. The residue was dissolved in 10 ml of dichloromethane and the solution was washed with 2 ml of saturated sodium hydrogen carbonate solution, dried over anhydrous magnesium sulfate and evaporated to give 36 mg of 2 (S) - [3 (S) amino-2 (R) -hydroxy -4-Phenylbutyl] -N-tert.butyl-1 (R) -cyclohexanecarboxamide as a colorless gum, MS m / e 346 [M] ⁺ , which was used without further purification.

Example 3

Treatment of 75 mg (0.15 mmol) of 2 (R) - [[4 (S) -benzyl-3-tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamidas p-toluenesulfonic acid in methanol, in an analogous manner to that described in the first paragraph of Example 1, gave 20 mg of 2 (R) - [3 (S) - (tert-butoxyformamido) -2 (R) hydroxy-4 -phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide; MS m / e 469 [M + Na] +, 447 [M + H] +.

2 (R) - [[4 (S) -benzyl-3-tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl] -Nertc.butyl-1 (R) -cyclohexanecarboxamide used as starting material , we prepared as follows:

(i) a solution of 18.40 g (0.10 mol) of cis-4 (R) -acetoxymethyl-5 (S) -hydroxymethyl) cyclohexene and 16.50 g (0.11 mol) of tert-butyldimethylsilyl chloride in 45 ml of dimethylformamide were stirred and cooled to 0 ° C. . 17.00 g (0.25 mol) of imidazole were added and the mixture was stirred at 0 ° C for one hour, then allowed to warm to room temperature and stirred for a further 160 minutes. The mixture was then poured into 400 ml of water and extracted with two 100 ml portions of diethyl ether. The combined extracts were washed with 200 ml of water, dried over anhydrous magnesium sulfate and evaporated to give 29.69 g of a straw colored liquid. The crude product was chromatographed on a silica gel column using 10% ethyl acetate / hexane for elution to give 20.85 g of cis-4 (R) (acetoxymethyl-5 (S) - [[(tert-butyl) (dimethyl) silyloxy] methyl ] -3cyclohexene as a colorless oil; MS m / e 299 [M + H] ⁺ .

(ii) 32.5 ml of 2M sodium hydroxide solution was added to a solution of 26.94 g (0.090 mol) of the product of paragraph (i) in a mixture of 90 ml of ethanol and 90 ml of tetrahydrofuran. The mixture was stirred for 80 minutes at room temperature and then the volatiles were removed by evaporation. The residue was diluted with 520 ml of water and extracted with three 150 ml portions of diethyl ether. The combined extracts were washed with 350 ml and 100 ml of water, dried over anhydrous magnesium sulfate and evaporated to give 22.77 g of cis-6 (S) - [[(tert-butyl) (dimethyl) silyloxy] methyl] -3-cyclohexenyl (R) -methanol as a colorless liquid; MS m / e 257 [M + H] ⁺ .

(iii) In the analogous manner as described in Example 1 (ii), 22.77 g (0.089 mol) of the product of paragraph (ii) was hydrogenated to give 19.85 g of cis-2 (S) [[(tert.butyl) (dimethyl) silyloxy] methyl] -1 (R) -cyclohexanmethanol as a colorless liquid; MS m / e 259 [M + H] +.

(iv) In an analogous manner to that described in Example 1 (iii), Swern oxidation of 23.15 g (0.090 mol) of the product of paragraph (iii) gave 22.78 g of cis-2 (S) [[(tert.butyl) ( dimethyl) silyloxy] methyl] -1 (R) -cyclohexanecarboxaldehydeacol colorless oil; MS m / e 257 [M + H] ⁺ .

(v) In an analogous manner to that described in Example 1 (iv), treatment of 17.70 g (0.069 mol) of the product of paragraph (iv) with methyl triphenylphosphonium bromide and n-butyllithium gave 14.51 g of cis-l (S) - [[(tert-butyl) (dimethyl) silyloxy] methyl] -2 (S) vinylcyclohexane as a colorless oil; MS m / e 255 [M + H] ⁺ .

(vi) In an analogous manner to that described in Example 1 (v), treatment of 12.47 g (0.049 mol) of the product of paragraph (v) with tetrabutylammonium fluoride gave 6.617 g of cis-2 (S) -vinyl-1 (S) ) -cyclohexanmethanol as a colorless oil; MS m / e 141 [M + H] +.

(vii) A solution of 6.56 g (47 mmol) of the product of paragraph (vi) in 30 ml of toluene was added over a 4 minute period to a stirred suspension of 22.56 g (53 mmol) of dibromotrophenylphosphorane in 120 ml of toluene. The mixture was stirred under nitrogen at room temperature for 20 hours, then filtered and the solid was washed with 200 ml of hexane. The combined filtrates were washed with two 200 ml portions of saturated sodium hydrogen carbonate solution and 200 ml water, dried over anhydrous magnesium sulfate and evaporated to give 7.898 g of cis-l (S) -bromomethyl-2 (S) -vinylcyclohexane as a colorless liquid ; MS m / e 205, 203 [M + H] ⁺ .

(viii) In an analogous manner to that described in Example 1 (viii), treatment of 7.766 g (38 mmol) of the product of paragraph (vii) with magnesium is followed by reaction with tert-butyl (La-formylphenethyl) carbamate, gave 3.196 g of 1 (R) - [3 (S) (tert-butoxyformamido) -2 (S) -hydroxy-4-phenylbutyl] -2 (S) -vinylcyclohexane containing 10% (R) -alcohol; MS m / e 374 [M + H] +.

(ix) In an analogous manner to that described in Example 1 (ix), oxidation of 2.81 g (7.5 mmol) of the product of paragraph (viii) with pyridinium dichromate gave 1.61 gl (R) - [3 (S) - (tert. butoxyformamido) -2-oxo-4-phenibutyl] -2 (S) -vinylcyclohexane as a colorless oil which crystallizes on condition; MS m / e 372 [M + H] ⁺ .

(x) In an analogous manner to that described in Example 1 (x), reduction of 1.44 g of the product of paragraph (x) with sodium borohydride gave 1.20 gl (R) - [3 (S) - (tert. butoxyformamido) - 2- (R) -hydroxy-4-phenibutyl] -2 (S) -vinylcyclohexane as a colorless gum which crystallizes on condition; MS m / e 373 [M] ⁺ .

(xi) In an analogous manner to that described in Example l (xi), treatment of 1.20 g (3.2 mmol) of the product of paragraph (x) with 2,2-dimethoxypropane gave 1.25 g of 1 (R) [[4 (S ) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5- (R) -oxazolidinyl] methyl] -2 (S) vinylcyclohexane as pale yellow gum; MS m / e 414 [M + H] ⁺ .

(xii) In an analogous manner to that described in Example 1 (xii), treatment of 850 mg (2.06 mmol) of the product of paragraph (xi) with potassium permanganate gave 185 mg of 2 (R) - [[4 (S) - benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5- (R) -oxazolidinyl] methyl] -1 (R) -cyclohexanecarboxylic acid as a colorless gum;

MS m / e 432 [M + H] +.

(xiii) In an analogous manner to that described in Example l (xiii), treatment of 165 mg (0.38 mmol) of the product of paragraph (xii) by coupling tert.butylamine gave 85 mg of 2 (R) - [[4 (S ) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5- (R) oxazolidinylmethyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide as a white solid; MS m / e 487 [M + H] ⁺ .

Example 4

In the analogous manner as described in the first paragraph of Example 2, we obtained from 2 (R) [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide and N-benzyloxycarbonyl-L-asparagine pentafluorophenyl ester 2 (R) [3 (S) - [[N- (benzyloxycarbonyl) -L-asparaginyl] -amino-2 (R) -hydroxy-4-phenylbutyl] N- tert.butyl-1 (R) -cyclohexane-carboxamide as a white solid from the ground. 166.5 to 168 ° C; MS m / e 617 [M + Na] + and 595 [M + H] +.

2 (R) - [3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide was used as starting material in an analogous manner as described in the second paragraph of Example 2, from 2 (R) - [3 (S) (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexane-carboxamide [obtained as described in the first paragraph of Example 3] and used without further purification.

Example 5

A mixture of 65 mg (0.13 mmol) 2 (S) - [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane- of carboxamide and 35 mg (0.13 mmol) of N-hydroxysuccinimide ester of quinaldic acid in 2 ml of tetrahydrofuran were stirred at room temperature under nitrogen for 18 hours. The solvent was removed by evaporation and the residue was partitioned between 10 ml of ethyl acetate and 10 ml of 10% sodium carbonate solution. The organic layer was washed with 10 ml of water, dried over anhydrous magnesium sulfate and evaporated. The crude product was chromatographed on a silica gel column using 7% methanol in dichloromethane for elution to give 80 mg of 2 (S) - [3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] amino] - 2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide as a white solid; MS m / e 616 [M + H] ⁺ .

2 (S) - [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexane-carboxamide used as starting material , we prepared as follows:

A solution of 80 mg (0.13 mmol) 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -Lasparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide [obtained as described in the first paragraph of Example 2] was hydrogenated in 10 ml of ethanol over 10 mg of 10% palladium-on-charcoal catalyst for 4 hours. The catalyst was removed by filtration and the filtrate was evaporated. The residue was suspended in toluene and the mixture was evaporated. This procedure was repeated once to give 65 mg of 2 (S) - [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexane-carboxamide as a colorless foam, which was used without further purification.

Example 6

A solution of 90 mg (0.26 mmol) of 2 (S) - [3 (S) -amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1 (R) -cyclohexane-carboxamide in 4 ml of dichloromethane a mixture of 65 mg (0.26 mmol) of N-benzyloxycarbonyl-cyano-L-alanine, 35 mg (0.23 mmol) of hydroxybenzotriazole hydrate and 54 mg (0.26 mmol) of dicyclohexylcarbodiimide in 2 ml of dimethylformamide were added. The mixture was stirred under nitrogen at room temperature for 18 hours and then filtered. The filtrate was evaporated and the residue partitioned between 25 ml of ethyl acetate and 10 ml of saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over anhydrous magnesium sulfate and evaporated. The crude product was chromatographed on a silica gel column using 3% methanol in dichloromethane for elution and the product was further purified by crystallization from a mixture of 1 ml of dichloromethane and 10 ml of hexane. This gave 68 mg of 2 (S) - [3 (S) [[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-t-butyl- l (R) -cyclohexane-carboxamide as a white solid;

MS m / e 577 [M + H] +.

Example 7

In an analogous manner to that described in Example 6, 73 mg (0.29 mmol) of N-benzyloxycarbonyl-L-valine were combined with 100 mg (0.29 mmol) of 2 (S) - [3 (S) -amino2 (R) -hydroxy- 4-Phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide to give 80 mg of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide as a white solid;

MS m / e 580 [M + H] ⁺ .

Example 8

In an analogous manner to that described in Example 6, 78 mg (0.26 mmol) of N-benzyloxycarbonyl-L-phenylalanine were combined with 90 mg (0.26 mmol) of 2 (S) - [3 (S) amino] -2 (R) - hydroxy-4-phenylbutyl-N-tert-butyl-1 (R) -cyclohexane-carboxamide to give 72 mg of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-phenylalanyl] amino ] 2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamidacate white solid; MS m / e 628 [M + H] < ⁺ >.

Example 9

In the analogous manner as described in Example 5, 95 mg (0.16 mmol) of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-valyl] amino] -2 (R) - hydroxy-4-phenylbutyl] -Nertc.butyl-1 (R) -cyclohexanecarboxamide was hydrogenated over 10% palladium-on-carbon catalyst and the product was reacted with quinaldic acid N-hydroxysuccinimide ester to give 50 mg of 2 (S) - [3 (S ) - [[N- (2-quinolylcarbonyl) -L-valyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide as a white solid; MS m / e 601 [M + H] ⁺ .

Example 10

In an analogous manner to that described in Example 6, 70 mg (0.26 mmol) of N-benzyloxycarbonyl-S-methyl-L-cysteine were combined with 90 mg (0.26 mmol) of 2 (S) - [3 (S) amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide to give 30 mg of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) - S-methyl-Lcysteinyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide as a white solid from m.p. 143-144 ° C; MS m / e 598 [M + H] < ⁺ >.

Example 11

220 mg (0.59 mmol) of pyridinium dichromate was added to a solution of 48 mg (0.08 mmol) of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-valyl] amino] -2 (R) -hydroxy4 -phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide in 1 ml dimethylformamide and the mixture was stirred for 18 hours at room temperature. 10 ml of water were added and the mixture was extracted with two 10 ml portions of ethyl acetate. The combined extracts were washed with two 10 ml portions of water, dried over anhydrous magnesium sulfate and evaporated. The crude product was chromatographed on a silica gel column using 4% methanol in dichloromethane for elution to give 31 mg of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) -L-valyl] amino] -2- oxo-4-phenylbutyl] -Nterc.butyl-1 (R) -cyclohexane-carboxamide as a white solid from m.p. 186-188 ° C; MS m / e 578 [M + H] ⁺ .

Example 12

In the analogous manner as described in Example 6, 72 mg (0.26 mmol) of N-benzyloxycarbonyl-L-aspartic acid β-methyl ester was combined with 90 mg (0.26 mmol) of 2 (S) - [3 (S) - amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexane-carboxamide to give 90 mg of 2 (S) - [3 (S) - [[N- ( benzyloxycarbonyl) -O-methyl-L-aspartyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexane-carboxamide as a colorless gum;

MS m / e 610 [M + H] +

Example 13

A mixture of 60 mg (0.13 mmol) 2 (S) - [3 (S) - [[L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane- of carboxamide, 25 mg (0.13 mmol) of 3,5 dichlorobenzoic acid, 18 mg (0.13 mmol) of hydroxybenzotriazole hydrate and 27 mg (0.13 mmol) of dicyclohexyl-carbodiimide in 2 ml of dimethylformamide were stirred for 20 hours at room temperature under nitrogen. The mixture was then filtered and the solid was washed with dichloromethane. The combined wash filtrate was evaporated to dryness and the residue was partitioned between 50 ml of ethyl acetate and 10 ml of saturated aqueous sodium hydrogen carbonate solution. The separated organic phase was dried over anhydrous magnesium sulfate and evaporated. The residue was chromatographed on silica gel using 5% methanol in dichloromethane for elution. The product was further purified by recrystallization from a mixture of 2 ml of dichloromethane and 10 ml of n-hexane to give 37 mg of 2 (S) - [3 (S) - [[N- (3,5-dichlorobenzoyl) -L-asparaginyl] amino2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamidacate white solid from m.p. 220 - 226 ° C.

Example 14

In the analogous manner as described in Example 6, 68 mg (0.26 mmol) of N-benzyloxycarbonyl-3-methyl-L-valine were combined with 90 mg (0.26 mmol) of 2 (S) - [3 (S) amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide to give 40 mg of 2 (S) - [3 (S) - [[N- (benzyloxycarbonyl) - 3-methyl-L-valyl] amino] 2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexane-carboxamide as a white solid from m.p. 90 ° C (decomposition); MS m / e 594 [M + H] ⁺ .

Example 15

A solution of 100 mg (0.23 mmol) of 2 (S) - [[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl 1 (R) -cyclohexanecarboxylic acid (obtained as described in Example 1) in 5 ml of diethyl ether was added to 5 ml of a 0.3 M solution of diazomethane in diethyl ether. The mixture was allowed to stand at room temperature overnight and then evaporated. The residue was dissolved in 3 ml of methanol and 4 mg of toluene-4-sulfonic acid was added. The mixture was allowed to stand at room temperature overnight and then evaporated to dryness. The residue was partitioned between 10 ml of dichloromethane and 3 ml of saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over anhydrous magnesium sulfate and evaporated to give 90 mg of rubber. The crude product was chromatographed on a silica gel column using hexane / ethyl acetate (2: 1, v / v) for elution to give methyl 2 (S) - [3 (S) (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -1 (R) -cyclohexanecarboxylate as a white solid from the ground. 127 - 128 ° C; MS m / e 406 [M + H] +.

The following Example illustrates a typical pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable acid addition salt of a basic compound of Formula I as an effective ingredient:

Example A

The aqueous solution of the active ingredient is sterile filtered and stirred by heating with a sterile solution of gelatin containing phenol as a preservative, using such quantities that 1 ml of the resulting solution contains 3 mg of effective ingredient, 150 mg of gelatin, 4.7 mg of phenol and distilled water ad 1 ml. The mixture is filled into ampoules of 1 ml capacity under aseptic conditions.

hifitj / jana

Claims (18)

Compounds of the general formula wherein R1 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aralkanoyl, heterocyclylcarbonyl, or a group of formula represents alkyl, cycloalkylalkyl or aralkyl; represents hydrogen and R ^ represents hydroxy, or R ^ and R ^ together represent oxo; R4 represents alkoxycarbonyl or alkylcarbamoyl; R ^ and R ^ together represent trimethylene or tetramethylene, optionally substituted by alkyl or on adjacent carbon atoms by tetramethylene; R4 represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aroyl, aralkanoyl, or heterocyclylcarbonyl; and R4 represents alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, cyanoalkyl, carbamoylalkyl, alkylthioalkyl, aicoxyalkyl or alkoxycarbonylalkyl, and the pharmaceutically acceptable acid addition salts of those compounds of formula I which are basic. Compounds according to claim 1, wherein it represents alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aralkanoyl or heterocyclylcarbonyl. Compounds according to claim 1 or 2, wherein R1 represents alkoxycarbonyl or an asupine of formula (i) in which it represents aralkoxycarbonyl, aroyl or heterocyclycarbonyl and R1 represents alkyl, aralkyl, cyanoalkyl, carbamoylalkyl, alkylthioalkyl or alkoxycarbonylalkyl. Compounds according to claim 3, wherein R1 represents tert-butoxycarbonyl or a group of formula (i) in which R1 represents benzyloxycarbonyl, 3,5-dichlorobenzoyl or 2quinolylcarbonyl and R1 represents isopropyl, tert-butyl, benzyl, cyanomethyl, carbamoylmethyl, methylthiomethyl or methoxycarbonylmethyl. Compounds according to any one of claims 1-4, wherein R- represents aralkyl. Compounds according to claim 5, wherein R- represents benzyl. Compounds according to any one of claims 1-6, wherein R 1 represents hydrogen and R ⁴ represents hydroxy. 8. Compounds according to any one of claims 1-7, wherein it represents methoxycarbonyl. Compounds according to any one of claims 1-7, wherein R 1 represents tert-butylcarbamoyl. Compounds according to any one of claims 1 to 9, wherein R 1 and R 5? together they represent unsubstituted tetramethylene. Compounds according to any one of claims 1 - 10, wherein R1 represents tert-butoxycarbonyl or a group of formula (i) in which R1 represents benzyloxycarbonyl, 3,5-dichlorobenzoyl or 2-quinolylcarbonyl and R1 represents isopropyl, tert-butyl , benzyl, cyanomethyl, carbamoylmethyl, methylthiomethyl or methoxycarbonylmethyl, represents benzyl, R ⁴ represents hydrogen and R ⁴ represents hydroxy, R ⁴ represents methoxycarbonyl or tert-butylcarbamoyl and and R? together they represent unsubstituted tetramethylene. 12. 2 (S) - [3 (S) - [[N- (2-quinolylcarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexane-carboxamide. A compound according to claim 1 selected from: 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -3-cyano-L-alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] N-tert-butyl-1 (R) cyclohexane-carboxamide, 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-t-butyl-1 (R) cyclohexane-carboxamide, 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -L-phenyl] alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1 (R) cyclohexane-carboxamide , 2 (S) - [3 (S) - [[N- (2-quinolylcarbonyl) -L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1 (R) cyclohexane- carboxamide, 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -S-methyl-L-cysteyl] amino] -2 (R) -hydroxy-4-phenylbutyl N-tert-butyl-1 (R) cyclohexane-carboxamide, 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -L-valyl] amino] -2-oxo-4-phenylbutyl] -N-tert-butyl (R) cyclohexane-carboxamide, 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -O-methyl-L-aspartyl] amino] -2 (R) -hydroxy-4-phenylbutyl-N-tert-butyl-1 (R) cyclohexane- carboxamide, 2 (S) - [3 (S) - [[N-3,5-dichlorobenzoyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-t-butyl-1 (R) cyc ] ohexane-carbok.samide, 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -3-methyl-L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] N-tert-butyl-1 (R) cyclohexane-carboxamide and methyl 2 (S) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -1- (R) cyclohexane-carboxylate. A compound according to claim 2 selected from: 2 (S) - [3 (S) - [[N-benzyloxycarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -Nert-butyl-1 (R) cyclohexane-carboxamide, 2 (S) - [3 (S) - (tert-butoxyl'ormamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl (R) cyclohexane-carboxamide, 2 (R) - [3 (S) - (tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl (R) cyclohexane-carboxamide and 2 (R) - [3 (S) - [[N-benzyloxycarbonyl) -L-asparaginyl] amino-2 (R) -hydroxy-4-phenylbutyl] -N tert-butyl-1 (R) cyclohexane-carboxamide. 15. Compounds of general formulas: wherein R ^ ^a represents alkoxycarbonyl or aryloxycarbonyl and R ⁷ , R ⁷ , r ⁴ , R ⁵ , R ⁶ , R ⁷ and R 4 have the meaning given in claim 1. Amino acid derivative according to any one of claims 1-14 for use as a therapeutically effective substance. Amino acid derivative according to any one of claims 1 - 14 for use in the prophylaxis and treatment of viral infections. 18. A process for the preparation of an amino acid derivative according to any one of claims 1-14, characterized in that the method comprises (a) for the preparation of a compound of formula 1 in which R1 represents alkoxycarbonyl or aralkoxycarbonyl, R ³ represents hydrogen and R ⁴ represents hydroxy , treatment of a compound of the general formula in which R ^a represents alkoxycarbonyl or aralkoxycarbonyl and R ^3, R 5, R 6 and in claim 1 of the meanings given above, with an acid, or (b) by reacting a compound of the general formula