KR100220987B1 - Amino acid derivatives and their use as anti-viral agents - Google Patents

Abstract

Pharmaceutically acceptable salts of the compounds of formula (I) and of the compounds of formula (I) which are basic may inhibit the aspartyl protease of viral origin and may be used as pharmaceutical forms for the prevention and treatment of viral infections. .

Wherein R ¹ is alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aralkanoyl, heterocyclylcarbonyl or a general formula

Wherein R ⁸ is alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aroyl, aralkanoyl or heterocyclylcarbonyl, and R ⁹ is alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, cyanoalkyl , Carbamoylalkyl, alkylthioalkyl, alkoxyalkyl or alkoxycarbonylalkyl), R ² is alkyl, cycloalkylalkyl or aralkyl, R ³ is hydrogen, R ⁴ represents hydroxy, or Or R ³ and R ⁴ together represent oxo, R ⁵ is alkoxycarbonyl or alkylcarbamoyl, and R ⁶ and R ⁷ together are trimethylene or tetra optionally substituted by alkyl or by tetramethylene on adjacent carbon atoms Methylene is shown.

These compounds can be prepared according to commonly known methods.

Description

Amino acid derivatives, preparation methods thereof, and pharmaceuticals containing the same

The present invention relates to amino acid derivatives. In particular, the present invention relates to pharmaceutically acceptable acid addition salts of amino acid derivatives of general formula (I) and compounds of general formula (I) which are basic.

Wherein R ¹ is alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aralkanoyl, heterocyclylcarbonyl or a general formula wherein R ⁸ is alkoxycarbonyl, alkoxycarbonyl, alkanoyl,

Aroyl, aralkanoyl or heterocyclylcarbonyl, R ⁹ is alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, cyanoalkyl, carbamoylalkyl, alkylthioalkyl, alkoxyalkyl or alkoxycarbonylalkyl) R ² is alkyl, cycloalkylalkyl or aralkyl, R ³ is hydrogen, R ⁴ represents hydroxy or R ³ and R ⁴ together represent oxo and R ⁵ is alkoxycarbonyl Or alkylcarbamoyl, R ⁶ and R ⁷ together represent trimethylene or tetramethylene optionally substituted by alkyl or by tetramethylene on adjacent carbon atoms.

The compounds and salts mentioned above are novel and have useful pharmacological properties. In particular, the compounds and salts inhibit aspartyl proteases of viral origin and can be used to prevent and treat viral infections, especially infections caused by HIV and other retroroid viruses.

It is an object of the present invention to provide compounds of general formula (I) and these compounds and salts for use as therapeutically active substances as described above and the salts themselves; Intermediate products used in the process; Medicaments containing these compounds and salts; The use of these compounds and salts for preventing or controlling diseases, in particular for preventing or treating viral infections; And the use of these compounds and salts for the manufacture of a medicament for preventing or treating a viral infection.

As used herein, alone or in combination, the term “alkyl” refers to a straight or branched chain alkyl containing up to eight, preferably up to four carbon atoms, for example methyl, ethyl, n-propyl, iso Propyl, n-butyl, isobutyl, secondary-butyl, tert-butyl, n-pentyl, n-hexyl and the like. The term "alkoxy", used alone or in combination, refers to an alkyl ether group, wherein the term "alkyl" is as described above, for example, methoxy, ethoxy, n-propoxy, isoprop Foxy, n-butoxy, isobutoxy, secondary-butoxy, tertiary-butoxy and the like. The term "cycloalkyl", used alone or in combination, refers to a cycloalkyl group having 3 to 8 carbon atoms, preferably 3 to 6 carbon atoms, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. have. The term “aralkyl” refers to one or more substituents selected from alkyl groups as described above, wherein one hydrogen atom is substituted by an aryl group, ie, alkyl, alkoxy, halogen, trifluoromethyl, hydroxy, nitro, amino, and the like. Optionally containing phenyl or naphthyl group. Examples of aralkyl groups include benzyl, 4-chlorobenzyl, 4-methoxybenzyl, 2-phenylethyl, 2-naphthylethyl and the like. The term "aralkyloxycarbonyl" denotes a group of the general formula aralkyl-O-(CO)-wherein the term "aralkyl" is as defined above. The term "alkanoyl" refers to an acyl group derived from alkanecarboxylic acid, for example, acetyl, propionyl, butyryl, valeryl, 4-methylvaleryl and the like. The term "aroyl" denotes a benzoyl or naphthoyl group optionally containing one or more substituents selected from alkyl, alkoxy, halogen, trifluoromethyl, hydroxy, nitro, amino and the like, for example benzoyl, p- Chlorobenzoyl, 3,5-dichlorobenzoyl, 1-naphthoyl and the like. The term “arkanoyl” refers to an acyl group derived from aryl-substituted alkancarboxylic acid, for example phenylacetyl, 3-phenylpropionyl (hydrocinnamoyl), 4-phenylbutyryl, (2-naphthyl) ) Acetyl, 4-chloro-, 4-amino-, or 4-methoxyhydrocinnamoyl. The term "heterocyclylcarbotil 'refers to the general formula-CO-Het [where Het contains one or more heteroatoms selected from nitrogen, oxygen and sulfur and is optionally substituted by halogen, alkyl, alkoxy, oxo or the like on one or more carbon atoms. Substituted and / or substituted by alkyl, aralkoxycarbonyl, alkanoyl, phenyl or phenylalkyl on one secondary nitrogen atom (i.e., -NH-) or on the oxydo on the tertiary nitrogen atom (i.e. Is a saturated, partially unsaturated, or aromatic monocyclic, bicyclic, or tricyclic heterocycle, optionally substituted by C. C. Examples of such Het groups include pyrrolidinyl, piperidinyl , Morpholinyl, thiomorpholinyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyridinyl, furyl, thienyl, triazolyl, oxazolyl, thiazolyl, phosphorus Aryl, quinolyl, isoquinolyl, tetrahydroquinolyl, 1,2,3,4-tetrahydroisoquinolyl, quinoxalinyl, β-carbolinyl, etc. The term "halogen" refers to fluorine, chlorine, Bromine and iodine.

Pharmaceutically acceptable acid addition salts include basic compounds of formula (I) and inorganic acids such as hydrohalic acid (eg hydrochloric acid and hydrobromic acid), sulfuric acid, nitric acid, phosphoric acid, etc., or organic acids such as acetic acid. , Citric acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid, p-toluenesulfonic acid and the like.

Compounds of general formula (I) contain at least three asymmetric carbon atoms and are therefore present in the form of optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates or mixtures of diastereomeric racemates . The present invention includes all of these forms within its scope.

One particular group of compounds of formula (I) includes compounds wherein R ⁸ is alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aralkanoyl or heterocyclylcarbonyl.

In the compounds of formula (I), R ¹ is preferably alkoxycarbonyl, in particular tert-butoxycarbonyl, or R ⁸ is aralkoxycarbonyl (especially benzyloxycarbonyl), aroyl (especially 3,5-dichlorobenzoyl) or heterocyclylcarbonyl (particularly 2-quinolylcarbonyl), R ⁹ is alkyl (particularly isopropyl or tert-butyl), aralkyl (particularly benzyl), cya A general formula of noalkyl (especially cyanomethyl), carbamoylalkyl (especially carbamoylmethyl), alylthioalkyl (especially methylthiomethyl) or alkoxycarbonylalkyl (especially methoxycarbonylmethyl) ) Group. R ² preferably denotes aralkyl, in particular benzyl. Preferably R ³ represents hydrogen and R ⁴ represents hydroxy. When R ⁵ represents alkoxycarbonyl, it is preferably methoxycarbonyl and when R ⁵ is alkylcarbaroyl, it is preferably tert-butyl carbamoyl. Preferably, R ⁶ and R ⁷ together represent tetramethylene unsubstituted.

From the foregoing paragraphs, particularly preferred compounds of formula (I) are those in which R ¹ is tert-butoxycarbonyl, or R ⁸ is benzyloxycarbonyl, 3,5-dichlorobenzoyl or 2-quinolylcarbonyl ^Is a group of formula (i) wherein ⁹ is isopropyl, tert-butyl, benzyl, cyanomethyl, carbamoylmethyl, methylthiomethyl or methoxycarbonylmethyl, R ² is benzyl, R ³ is hydrogen It will be appreciated that R ⁴ is hydroxy, R ⁵ is methoxycarbonyl or tert-butylcarbamoyl, and R ⁶ and R ⁷ together represent an unsubstituted tetramethylene.

One particularly preferred compound of formula (I) is 2 (S)-[3 (S)-[[N-(2-quinolylcarbonyl)-L-asparaginyl] amino]-2 (R)- Hydroxy-4-phenylphenyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide.

Other particularly preferred compounds of formula (I) are as follows.

Other interesting compounds of formula (I) are illustrated below:

According to the process provided by the present invention, (a) to prepare a compound of formula (I) wherein R ¹ is alkoxycarbonyl or alkoxycarbonyl, R ³ is hydrogen and R ⁴ is hydroxy, Treating the compound of II) with an acid, (b) reacting the compound of formula (III) with an acylating agent which introduces a group R ¹ as defined above, or (c) R ¹ is To prepare a compound of formula (I) which is a group of, the compound of formula (IV) is reacted with an acylating agent which introduces group R ⁸ as defined above, or (d) R ³ and R ⁴ are To prepare a compound of formula (I) which together represent oxo, oxidize a compound of formula (I) wherein R ³ is hydrogen and R ⁴ is hydroxy, and / or (e) optionally Mixtures of isomeric racemates with diastereomeric racemates or optically pure diasteres Separating into isomers and / or (f) optionally separating the mixture of diastereomers into optically pure diastereomers, and / or (g) optionally Compounds of formula (I) and pharmaceutically acceptable acid addition salts of these compounds are prepared by converting the basic compounds into pharmaceutically acceptable acid addition salts.

Wherein R ^1a is alkoxycarbonyl or aralkoxycarbonyl, and R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ⁹ are as defined above.

When the compound of formula (II) is treated with an acid according to the aspect of method (a), a compound of formula (I) wherein R ¹ is alkoxycarbonyl or alkoxycarbonyl, R ³ is hydrogen and R4 is hydroxy Is obtained. The form of the acid used in this embodiment depends mainly on the nature of the substituent R ^1a present in the starting material of general formula (II). When R ^1a represents alkoxycarbonyl, eg tert-butoxycarbonyl, the treatment is carried out in the presence of an organic solvent inert under the reaction conditions, for example an alkanol (eg methanol, ethanol, etc.). Strong acids, in particular organosulfonic acids, for example alkanesulfonic acids (e.g. methanesulfonic acid, etc.), or aromatic sulfonic acids (e.g. benzenesulfonic acid, p-toluenesulfonic acid, mesitylenesulfonic acid, etc.) It is desirable to. However, halogenated alkancarboxylic acids such as trifluoroacetic acid can be used instead of organic sulfonic acids. When R ^1a is aralkoxy carbonyl, eg benzyloxycarbonyl, the treatment is carried out in the presence of an alkanol (eg, in the presence of an organic solvent inert under the reaction conditions, for example a halogenated aliphatic hydrocarbon such as dichloromethane, etc.) Methanol), preferably with hydrogen halide such as hydrogen chloride.

The process of reacting a compound of formula (III) with an acylating agent according to the embodiment of method (b) can be carried out in a manner known per se, using the corresponding acid or reactive derivative thereof as the acylating agent. Suitable reactive derivatives include acid halides (eg acid chlorides), acid anhydrides, mixed acid anhydrides, activated esters, and the like. When the acylating agent is one which introduces a group of the general formula, the reaction is condensed [eg; Dicyclohexylcarbodiimide or benzotriazole-1 -yloxy-tris (dimethylamino) phosphonium hexa fluorophosphate] and an acid in the presence of a base (e.g. triethylamine, ethyldiisopropylamine, etc.) It is convenient to carry by. Reaction about 0 Preference is given to performing at about room temperature, preferably at room temperature.

The process of reacting a compound of formula (IV) with an acylating agent according to the embodiment of process (c) can be carried out by a method known per se, using the corresponding acid or reactive derivative thereof as the acylating agent. Suitable reactive derivatives include acid halides (eg acid chlorides), acid anhydrides, mixed acid anhydrides, activated esters, and the like. Reaction about 0 Suitably at about room temperature, preferably at room temperature.

The oxidation according to the aspect of process (d) can be carried out in a manner known per se to oxidize the secondary alcohols to ketones. Thus, for example, pyridinium dichromate in dimethylformamide, pyridinium chlorochromate in dichloromethane, sulfur trioxide-pyridine complex in dimethyl sulfoxide, oxalyl chloride in dimethyl sulfoxide and triethylamine, dimethyl sulfoxide in Oxidation can be carried out using dicyclohexylcarbodiimide and organic acids such as dichloroacetic acid or trifluoroacetic acid.

Any separation process according to the embodiments of methods (e) and (f) can be carried out according to conventional methods such as, for example, column chromatography, thin layer chromatography, high pressure liquid chromatography and the like.

The process for converting the basic compound of general formula (I) into a pharmaceutically acceptable salt according to the embodiment of method (g) can be carried out using inorganic acids such as hydrohalic acid (eg hydrochloric acid or hydrobromic acid), sulfuric acid, nitric acid, Phosphoric acid and the like, or organic acids such as acetic acid, citric acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid, p-toluenesulfonic acid, and the like, can be carried out by conventional methods.

The compounds of formula (II) used as starting materials in the embodiments of process (a) are novel and form another object of the present invention. The compound can be prepared according to the following scheme (wherein R ^1a , R ² , R ⁶ and R ⁷ are as defined above).

In relation to the above reaction scheme, in the first step, the compound of formula (V) is reacted with the compound of formula (VI) to obtain a compound of formula (VII). The reaction is about 0 in an organic solvent, such as ether (eg diethyl ether), which is inert under conventional Grignard reaction conditions, for example under reaction conditions. To about 40 , Preferably at approximately room temperature.

In the next step, the compound of formula IV is reacted with 2,2-dimethoxypropane in the presence of a strong organic acid, preferably organic sulfonic acid (e.g. p-toluenesulfonic acid). Switch to The reaction is conveniently carried out at approximately room temperature.

Subsequently, the compound of formula IV is oxidized to obtain a compound of formula IV. The oxidation is preferably carried out at about room temperature using an alkali metal permanganate such as potassium permanganate. Conveniently, oxidation comprises a mixture of water, alkancarboxylic acids (e.g. glacial acetic acid) and inert organic solvents that are not miscible with them, for example aromatic hydrocarbons (e.g. benzene, toluene, etc.) in the presence of a phase transfer catalyst. It is performed in a solvent system.

Finally, the compound of formula (VII) is converted to the compound of formula (II) by esterification or amidation. Esterification or amidation is carried out by reacting a compound of general formula with a suitable alkanol or amine according to methods known per se.

Compounds of formulas (V) and (VI) used to prepare compounds of formula (II) are known compounds or homologues of known compounds which can be prepared by analogous methods to known compounds. In addition, the examples described below describe details in connection with the preparation of specific compounds of formula (VI). In addition, the compound of general formula (i), (iv), and (iv) is new and corresponds to the objective of this invention.

The compounds of formula (III) used as starting materials in the embodiments of process (b) are novel and also form the object of the present invention. The compound may be prepared by decomposing and removing an alkoxycarbonyl or alkoxycarbonyl group from the compound of formula (I) wherein R ¹ is an alkoxycarbonyl or alkoxycarbonyl group. Decomposition can be carried out according to a method known per se. For example, when R ¹ is alkoxycarbonyl, the decomposition may be conveniently performed using a strong inorganic acid (e.g. hydrohalic acid) or a strong organic acid (e.g. trifluoroacetic acid). To about room temperature, wherein when R ¹ is an alkoxycarbonyl group, an organic solvent inert under the reaction conditions, such as an alkanol (eg ethanol or the like) or an alkancarboxylic acid ester (eg ethyl acetate) Among others, the decomposition can be conveniently carried out using hydrogen in the presence of a noble metal catalyst (eg palladium / charcoal) at approximately room temperature.

The compounds of formula (IV) used as starting materials in embodiments of process (c) are novel and form another object of the present invention. The compound can be prepared by decomposing an alkoxycarbonyl or alkoxycarbonyl group from the compound of formula (I) wherein R ¹ is a group of formula (i) and R ⁸ is an alkoxycarbonyl or alkoxycarbonyl group. Can be. The decomposition can be carried out according to a method known per se. For example, when R ⁸ is an alkoxycarbonyl group, it is conveniently about 0 using strong inorganic acids (eg hydrohalic acid) or strong acid organic acids (eg trifluoroacetic acid). Decomposition may be carried out at about room temperature, and if R ⁸ is an araloxycarbonyl group, an organic solvent, such as alkanol, which is inert under the reaction conditions in the presence of a noble metal catalyst (eg palladium / charcoal) In (e.g. ethanol or the like) or alkancarboxylic acid esters (e.g. ethyl acetate), the decomposition can be conveniently carried out using hydrogen at approximately room temperature.

As mentioned above, the compounds of general formula (I) and pharmaceutically acceptable acid addition salts of these compounds which are basic, inhibit aspartyl proteases of viral origin, and are infected with viral infections, in particular HIV and other retroid viruses It is useful for the treatment and prevention of infection caused by.

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

HIV protease is expressed in E. coli and then partially purified from soluble extracts of bacteria by ammonium sulfate fractionation (0-30%), heptapeptide succinyl protected as substrate-Val-Ser Protease activity is assayed using Gln-Asn-Phe-Pro-Ile isobutylamide. The degree of degradation of the substrate is quantified by measuring the amount of H-Pro-Ile isobutylamide produced by spectroscopic assay of N-terminal proline.

1.25 mM of the substrate is dissolved in 125 mM citric acid buffer (pH 5.5) containing 0.125 mg / ml Tween 20. 10 μl of various test compound solutions (dissolved in methanol or dimethyl sulfoxide and then diluted with water containing 0.1% Tween 20) and 10 μl of protease are added to 80 μl of the above buffered substrate. 37 The digestion was terminated by addition of 1 ml of colorant [30 μg / ml of isatin in ethanol and 1.5 mg / ml (volume / volume) of 2-(4-chlorobenzoyl) benzoic acid in 10% acetone) at. The solution is heated in a water bath and the colored residue is redissolved in 1 ml of 1% pyrogallol (weight / volume / volume) in 33% water in acetone. The optical density of the solution is measured using spectrophotometry at 599 nm. Comparison of the H-Pro-Ile isobutylamide prepared in the presence of the test compound with the control sample and the concentration of the test compound (I ₅₀ ) required to inhibit 50% using a graph showing the test compound at various concentrations tested Decide

In vitro antiviral activity of compounds of formula (I) can be demonstrated through the assays described below.

The assay uses HTLV-111 (strain RF) grown in C8166 cells (human CD4 ⁺ T lymphoplastoid strain) using RPM1 1640 containing bicarbonate buffer, antibiotics and 10% calf fetal serum.

Cell suspensions were infected 10 times with TCID ₅₀ virus, and then 37 Adsorb for 90 minutes at. The cells are washed three times with medium. The test is carried out in 6 ml of tissue culture tubes containing 2 x 10 ⁵ infected cells in 1.5 ml of medium. The test compound is dissolved in aqueous medium or dimethyl sulfoxide depending on solubility, and then 15 μl of a solution of test compound is added. Place the culture in a humid atmosphere containing 5% carbon dioxide in air. Incubate for 72 hours at. Subsequently, the culture was centrifuged, an aliquot of the supernatant was dissolved in Nonidet P40, and then a primary antiserum and horseradish peroxidase detection system exhibiting specific reactivity to the viral p24 antigen. Assay by the antigen capture assay used. The degree of coloring is measured using spectrophotometry and plotted against the concentration of the test compound. Determine the concentration (I ₅₀ ) required to protect 50%.

The results obtained in the above test using representative compounds of general formula (I) are indicated in the following table.

NT = Not tested

Pharmaceutically acceptable salts of compounds of formula (I) and compounds of formula (I) which are basic may be used as medicaments, for example in the form of pharmaceutical preparations. Pharmaceutical formulations may be administered orally in the form of enteric administration, eg, tablets, tablets, dragees, hard and soft gelatin capsules, solutions, emulsions or suspensions, intranasally in the form of nasal sprays, or in the form of suppositories. It may be administered rectally or parenterally, for example intramuscularly or intravenously in the form of injection solutions.

To prepare pharmaceutical formulations, the compounds and salts described above can be processed with therapeutically inert inorganic or organic excipients. Lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof can be used, for example, as excipients for tablets, coated tablets, dragees and hard gelatin capsules. Suitable excipients for soft gelatin capsules include, for example, vegetable oils, waxes, fats, semisolid and liquid polyols and the like. However, depending on the nature of the active ingredient, excipients may not normally be required in the case of soft gelatin capsules. Suitable excipients for preparing solutions and syrups are, for example, water, polyols, saccharose, invert sugar, glucose and the like. Suitable excipients for preparing injectable solutions are, for example, water, alcohols, polyols, glycerin, vegetable oils and the like. Natural hardened oils, waxes, fats, semi-liquid polyols and the like are suitable excipients for preparing suppositories.

Pharmaceutical formulations may also contain preservatives, stabilizers, wetting agents, emulsifiers, sweeteners, coloring agents, flavoring agents, salts for controlling osmotic pressure, buffers, skinning agents or antioxidants. In addition, the formulations may contain other therapeutically active substances.

Pharmaceuticals containing a pharmaceutically acceptable salt of a compound of general formula (I) and a basic compound of general formula (I) with a therapeutically inert excipient, and methods for preparing such agents are also objects of the present invention. do. The method for the preparation of a medicament is characterized in that the compound of formula (I) or a salt thereof is formulated in a herbal dosage form with a therapeutically inert excipient and, if necessary, one or more other therapeutically active substances. .

As mentioned above, the compounds of formula (I) and their salts described above can be used for the control or prevention of diseases, in particular for the prevention or treatment of viral infections (especially rat viral infection). Dosages can vary within wide limits and are, of course, adjusted to the individual needs in each particular case. Typically, for oral administration, the daily dosage is preferably from about 3 mg to about 3 g, preferably from about 10 mg to about 1 g, although it may be exceeded if it is found appropriate. The daily dose may be administered once or in several doses.

The following examples illustrate the invention.

Example 1

2 (S)-[[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl] in 3 ml of methanol] -N-3 A solution of 100 mg (0.20 mmol) of tert-butyl-1 (R) -cyclohexanecarboxamide and 3.3 mg (0.017 mmol) of p-toluene sulfonic acid is maintained at room temperature for 40 hours. The solvent is removed by evaporation and the residue is partitioned between 10 ml of dichloromethane and 2 ml of saturated sodium bicarbonate solution. The organic solution is dried over anhydrous magnesium sulfate and evaporated to yield 90 mg of rubber. The crude product was chromatographed on silica gel column with 33% ethyl acetate / hexane as eluent to give 2 (S)-[3 (S)-(tert-butoxyformamido) -2 (R) -hydroxy Phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide 45 mg is obtained; MS m / e 447 [M + H] ⁺ .

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

(Iii) A solution of 4.375 g (31 mmol) of trans-4 -cyclohexene-1 (R), 2 (R) -dimethanol and 4.66 g (31 mmol) of tert-butyldimethylsilyl chloride in 16 ml of dimethylformamide under nitrogen. After stirring, 0 Cool to 5.30 g of imidazole are added and the mixture is allowed to warm to room temperature and then stirred overnight. 200 ml of water are added and the mixture is extracted three times with 100 ml fractions of diethyl ether. The combined extracts are 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 as eluent to trans-1 (R), 2 (R) -bis-[[(tert-butyl) (dimethyl) silyloxy] methyl] 2.80 g of cyclohexene are obtained as a colorless oil. Eluting again through column to give 2.576 g of trans-6 (R)-[[(tert-butyl) (dimethyl) silyloxy] methyl] -3-cyclohexene-1 (R) -methanol as colorless oil; MS m / e 257 [M + H] ⁺ .

(Ii) A solution of 2.57 g (10 mmol) of the second product of short in 50 ml of ethanol is hydrogenated on 130 mg of 10% palladium / carbon catalyst for 1 hour at room temperature under atmospheric pressure. Filtration removes the catalyst and the filtrate is evaporated to yield 2.30 g of trans-2 (R)-[[(tert-butyl) (dimethyl) silyloxy] methyl] -1 (R) -cyclohexanemethanol as 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 then-78 Cool to A solution of 1.3 ml (1.43 g, 18 mmol) of anhydrous dimethyl sulfoxide in 5 ml of dichloromethane is added over 6 minutes. The mixture is stirred for a further 2 minutes, then a solution of 2.30 g (8.9 mmol) of the product from paragraph (ii) in 10 ml of dichloromethane is added over 9 minutes. Mixture-70 After stirring for 20 minutes, 5.5 ml (3.99 g, 39 mmol) of triethyl amine are added over 6 minutes. The mixture is allowed to warm to room temperature and then 40 ml of water are added. The phases are separated and the aqueous solution is extracted twice with 50 ml fractions of dichloromethane. The combined organic solutions are dried over anhydrous magnesium sulfate and evaporated to yield 2.45 g of colorless oil. This was purified by chromatography on a silica gel column using 2.5% ethyl acetate in hexane as eluent to give trans-2 (R)-[[(tert-butyl) (dimethyl) silyloxy] methyl] -1 (R) 1.53 g of cyclohexanecarboxaldehyde are obtained as a colorless rubber; MS m / e 257 [M + H] ⁺ .

(Iii) 3.46 ml (8.6 mmol) of n-butyllithium 2.5 M solution in hexane are added to a stirred suspension of 3.08 g (8.6 mmol) of methyl triphenylphosphonium bromide in 23 ml of diethyl ether over 5 minutes. The mixture was stirred at room temperature for 85 minutes, then a solution of 1.53 g (6.0 mmol) of the short product in 10 ml of diethyl ether was added over 5 minutes. The mixture is stirred for 140 minutes at room temperature, 20 ml of water is added and the phases are separated. The aqueous solution is extracted twice with 20 ml fractions of diethyl ether and the combined organic solutions are washed with 20 ml water and dried over anhydrous magnesium sulfate and then evaporated to yield 1.85 g of oil. This was purified by chromatography on a silica gel column using 1% ethyl acetate in hexane as eluent, to give trans-1 (R)-[[(tert-butyl) (dimethyl) silyloxy] methyl] -2 (S) 0.870 g of vinylcyclohexane is obtained as a colorless oil; MS m / e 239 [M-CH ₃ ] ⁺ .

(Iii) A solution of 870 mg (3.4 mmol) of the short product in 6 ml of a 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran is maintained at room temperature for 3.5 hours. The solvent is then evaporated and the residue is dissolved in 30 ml of water and then extracted three times with 20 ml portions of diethyl ether. The combined extracts are dried over anhydrous magnesium sulfate and then evaporated to yield 910 mg of colorless oil. This was chromatographed on a silica gel column with 20% ethyl acetate in hexane as eluent to give 435 mg of trans-2 (S) -vinyl-1 (R) -cyclohexanemethanol as colorless oil; MS m / e 141 [M + H] ⁺ .

(Iii) A solution of 435 mg (3.1 mmol) of the short-circuit product in 10 ml of pyridine is stirred under nitrogen and then cooled in an ice bath. 265 μl (391 mg, 3.4 mmol) of methane sulfonyl chloride are added, the ice bath is removed and the mixture is stirred at room temperature for 4 hours. The solvent is removed by evaporation and the residue is partitioned between 30 ml of 2M hydrochloric acid and 30 ml of diethyl ether. The aqueous solution was extracted twice with 30 ml fractions of diethyl ether, and the combined extracts were 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 then evaporated to give 1 (R)-[(methane Sulfonyloxy) methyl] -2 (S) -vinyl cyclohexane 615 mg are obtained as a colorless oil; MS m / e 219 [M + H] ⁺ .

(Iii) A mixture of 1.55 g (7.1 mmol) of short-circuit product and 935 mg (10.7 mmol) of lithium bromide in 20 ml of dimethylformamide was added under nitrogen. Stir for 50 hours. The mixture is then poured into 250 ml of water and extracted three times with 100 ml fractions of diethyl ether. The combined extracts are washed with 200 ml of water, dried over anhydrous magnesium sulfate and evaporated to give 1.312 g of trans-1 (R) -bromomethyl-2 (S) -vinyl cyclohexane as a colorless liquid; MS m / e 123 [M-Br] ⁺ .

(Iii) A mixture of 172 mg (7.1 mmol) of magnesium flakes and iodine crystals in 2 ml of tetrahydrofuran is stirred under argon, followed by addition of 1.113 g (5.5 mmol) of the short product over 3 minutes. The mixture is stirred and refluxed for 1 hour and then cooled to room temperature. A solution of 683 mg (2.74 mmol) of tert-butyl (L-α-formylphenethyl) carbamate in 5 ml of tetrahydrofuran is added over 10 minutes and the mixture is stirred at room temperature for an additional 2.5 hours. 30 ml of 10% ammonium chloride solution is then added, the pH is adjusted to 2 with 2M hydrochloric acid, and then extracted three times with 25 ml fractions of ethyl acetate. The combined extracts are washed with 25 ml of 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 as eluent to give 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]- 630 mg of a mixture of 2 (S) -vinylcyclo hexane are obtained as a white crystalline solid; MS m / e 374 [M + H] ⁺ . The diastereomeric mixture is used in the next step without further purification.

(Iii) A solution of 630 mg (1.7 mmol) of the short-circuit product in 20 ml of dimethylformamide was stirred and then 0 Cool to 4.48 g (12 mmol) of pyridinium dichromate was added and the mixture was Stir for 10 minutes. The cooling bath is then removed and the mixture is stirred at room temperature for 6 hours and then poured into 170 ml of water. The resulting mixture is extracted three times with 80 ml fractions of ethyl acetate, the combined extracts are washed with 100 ml of water, dried over anhydrous magnesium sulfate and then evaporated to give 1 (S)-[3 (S)-(tertiary -parts). Oxyformamido) -2-oxo-4-phenylbutyl]-2 (S)-vinylcyclohexane 640 mg is obtained as a colorless oil which crystallizes when left standing. The product is used in the next step without further purification.

(Iii) a solution of 640 mg (1.7 mmol) of the short-circuit product in 35 ml of ethane was After stirring, 368 mg (9.7 mmol) of sodium borohydride are added. Mixture 0 After stirring for 2.5 hours, the solvent is evaporated to remove the solvent. The residue is partitioned between 100 ml of water and 100 ml of ethyl acetate, the phases are separated and the aqueous phase is extracted with 100 ml fractions and 50 ml fractions of ethyl acetate, respectively. The combined extracts are dried over anhydrous magnesium sulfate and evaporated to give 595 mg of a white oily solid. This was chromatographed on a silica gel column using 20% ethyl acetate / hexane as eluent to give 1 (S)-[3 (S)-(tert-butoxyformamido) -2 (S) -hydroxy-4 -Phenylbutyl]-2 (S)-85 mg of vinylcyclohexane as a white waxy solid; MS m / e 374 [M + H] ⁺ . Elution was carried out again through the column to give 1 (S)-[3 (S)-(tert-butoxyformamido)-2 (R)-hydroxy-4-phenylbutyl]-2 (S)-vinylcyclohexane 263 mg are obtained as a white solid; MS m / e 374 [M + H] ⁺ .

(xi) A solution of 370 mg (0.99 mmol) of the second product of short in 7 ml of 2,2-dimethoxypropane and 225 mg (1.2 mmol) of p-toluenesulfonic acid monohydrate is allowed to stand overnight at room temperature. The solution is diluted with 45 ml of diethyl ether, washed twice with 40 ml fractions of sodium hydrogen carbonate, dried over anhydrous magnesium sulphate and evaporated to give 530 mg of a yellow oil. This was chromatographed on a silica gel column with 5% ethyl acetate / hexane as eluent to give 1 (S)-[[4- (S) -benzyl-3- (tert-butoxycarbonyl) -2,2- Dimethyl-5 (R) -oxazolidinyl] methyl] -2 (S) -vinylcyclohexane 230 mg is obtained 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 is 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 acetic acid in 8 ml of benzene. The mixture is stirred vigorously overnight at room temperature, then 428 mg of sodium metabisulfite is added. 24 ml of diethyl ether and 2.5 ml of 2M citric acid are added and the mixture is stirred for 10 minutes and then filtered. The phases are separated and the aqueous phase is extracted twice with 10 ml fractions of diethyl ether. The combined extracts are washed twice with 10 ml of water each time, 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 / dichloromethanol as eluent to give 2 (S)-[[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl- 210 mg of 5 (R) -oxazolidinyl] methyl] -1 (R) -cyclohexanecarboxylic acid is obtained as a colorless rubber, which is used in the next step without further purification.

(xiii) 210 mg (0.49 mmol) of the product of paragraph (xii) in 2 ml of dimethylformamide, 67 mg (0.49 mmol) of 1-hydroxybenzotriazole hydrate, 101 mg (0 49 mmol) of dicyclohexylcarbodiimide and tert-butyl A mixture of 52 μl (36 mg, 0.49 mmol) of amine is stirred at room temperature under nitrogen for 20 hours. The mixture is filtered and the solid is washed with 2 ml of ethyl acetate. The combined filtrates are evaporated and the residue is partitioned between 10 ml of ethyl acetate and 10 ml of saturated sodium bicarbonate solution. The phases are separated and the aqueous phase is extracted twice with 10 ml fractions of ethyl acetate. The combined extracts are washed with 10 ml of saturated sodium chloride solution, dried over anhydrous magnesium sulfate and evaporated to give 280 mg of an oily solid. It is extracted twice with 1 ml fraction of diethyl ether and then discarded. The ether solution is evaporated to give 260 mg of crude product. This was chromatographed on a silica gel column using 22% ethyl acetate / hexane as eluent to give 2 (S)-[[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl -109 mg of 5 (R) -oxazolidinyl] methyl] -N-tert-butyl- 1 (R) -cyclohexanecarboxamide as a white solid; MS m / e 487 [M + H] ⁺ .

Example 2

35 mg of 2 (S)-[3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide in 1 ml dioxane 0.1 mmol) and a mixture of 44 mg (0.1 mmol) of N-benzyloxycarbonyl-L-asparagine pentafluorophenyl ester are stirred under nitrogen at room temperature for 16 hours. The solvent is evaporated and the residue is dissolved in 10 ml of ethyl acetate, The solution is washed twice with 3 ml fractions of 2 M hydrochloric acid, once with 3 ml 2 M sodium hydroxide solution and once with 3 ml saturated sodium chloride solution, then dried over anhydrous magnesium sulfate and evaporated. The residue is chromatographed on a silica gel column with 6% methanol / dichloromethane as eluent to afford 34 g of a white solid. It was recrystallized from dichloromethane / hexanes to give 2 (S)-[3 (S)-[[N-(benzyloxycarbonyl)-L-asparaginyl] amino]-2 (R)-hydroxy-4- Phenylbutyl] -N-tert-Butyl-1 (R) -cyclohexanecarboxamide 14mg, melting point 195-199 Obtained as a phosphorus white solid; MS m / e 595 [M + H] ⁺ .

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

2 (S)-[3 (S)-(tert-butoxyformamido)-2 (R)-hydroxy-4-phenylbutyl]-N-tertiary-butyl-1 (R)-cyclohexane 45 mg (0.1 mmol) of carboxamide (obtained as described in the first paragraph of Example 1) were dissolved in 2 ml of ethyl acetate and the solution was Cool to 0.2 ml of saturated solution of hydrogen chloride in ethyl acetate is added and the mixture is allowed to stand at room temperature overnight. Then 1.0 ml of hydrogen chloride in ethyl acetate are added and the mixture is stirred for an additional 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 bicarbonate solution, then dried over anhydrous magnesium sulfate and evaporated to give 2 (S)-[3 (S) -amino-2 (R) -hydroxy- 4-mg of phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide was obtained as a colorless rubber (MS m / e 346 [M] ⁺ ), which was used without further purification. .

Example 3

In a manner similar to that described in the first paragraph of Example 1, 2 (R)-[[4 (S) -benzyl-3-tert-butoxycarbonyl) using p-toluenesulfonic acid in methanol 2,2-Dimethyl-5 (R) -oxazolidinyl] methyl] N-tert-Butyl-1 (R) cyclohexanecarboxamide 75 mg (0,15 mmol) was treated to give 2 (R)- [3 (S)-(tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) cyclohexanecarboxamide 20mg To obtain; MS m / e 469 [M + Na] ⁺ , 447 [M + H] ⁺ .

2 (S)-[[4 (S) -benzyl-3- (tert-butoxycarbonyl) -2,2-dimethyl5 (R) -oxazolidinyl] methyl] used as starting material] -N- Tert-Butyl-1 (R) -cyclohexanecarboxamide is prepared as follows:

(i) 18.40 g (0.10 mo1) of cis-4 (R) -acetoxymethyl-5 (5) -hydroxymethyl) cyclohexene and 16.50 g (0.11 mol) of tert-butyldimethylsilyl chloride in 45 ml of dimethylformamide After stirring the solution of 0 Cool to 17.00 g (0.25 mo1) of imidazole was added and the mixture was After stirring for 1 hour at room temperature and allowed to warm to room temperature, it is stirred for an additional 160 minutes. The mixture is then poured into 400 ml of water and extracted twice with 100 ml fractions of diethyl ether. The combined extracts are washed with 200 ml of water, dried over anhydrous magnesium sulfate and evaporated to yield 29.69 g of a pale yellow liquid. The crude product was chromatographed on silica gel column with 10% ethyl acetate / hexane as eluent to give cis-4 (R)-(acetoxymethyl) -5 (S)-[(tert-butyl) (dimethyl) silyl 20.85 g of oxy] methyl-3 cyclohexane are obtained as a colorless oil; MS m / e 299 [M + H] ⁺ .

(ii) 32.5 ml of 2M sodium hydroxide solution is added to a solution of 26.94 g (0.090 mol) of the product of paragraph (i) in a mixture of 90 ml of ethane and 90 ml of tetrahydrofuran. The mixture is stirred at room temperature for 80 minutes and then evaporated to remove volatile solvents. The residue is diluted with 520 ml of water and then extracted three times with 150 ml fractions of diethyl ether. The combined extracts were washed with 350 ml and 100 ml of water, dried over anhydrous magnesium sulfate and then evaporated to cis-6 (S)-[[(tert-butyl) (dimethyl) silyloxy] methyl-3 -cyclohexene-1 (R)-methanol is obtained as a colorless liquid; MS m / e 257 [M + H] ⁺ .

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

(iv) In a manner similar to that described in Example 1 (iii), 23.15 g (0.090 mo1) of the product of paragraph (iii) was swerned to give cis-2 (S)-[[class (3) Butyl) (dimethyl) silyloxy] methyl] -1 (R) -22.78 g of cyclohexane carboxaldehyde are obtained as a colorless oil; MS m / e 257 [M + H] ⁺ .

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

(vi) In a manner similar to that described in Example 1 (v), 12.47 g (0.049 mol) of the product of paragraph (v) were treated with tetrabutyl ammonium fluoride to give cis-2 (S) -vinyl-1 ( S)-6.617 g of cyclohexanemethane are obtained 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 is added to a stirred suspension of 22.56 g (53 mmol) of dibromotriphenylphosphorane in 120 ml of toluene over 4 minutes. The mixture is stirred at room temperature under nitrogen for 20 hours, then filtered and the solid is washed with 200 ml of hexane. The combined filtrates were washed twice with 200 ml fractions of saturated sodium hydroxide solution and 200 ml of water, dried over anhydrous magnesium sulfate and then evaporated to cis-1 (S) -bromomethyl-2 (S) -vinylcyclohexane 7.898 g is obtained as a colorless liquid; MS m / e 205, 203 [M + H] ⁺ .

(viii) In a manner similar to that described in Example 1 (viii), 7.766 g (38 mmol) of the product of paragraph (vii) were treated with magnesium and then tert-butyl (L-α-formylphenethyl) carba Reacted with a mate (R)-1 (R) containing 10% alcohol-[3 (S)-(tert-butoxyformamido)-2 (S)-hydroxy-4-phenylbutyl]- 2 (S)-3.196 g of vinylcyclohexane is obtained; MS m / e 374 [M + H] ⁺ .

(ix) In a manner similar to that described in Example 1 (ix), 2.81 g (7.5 mmol) of the product of paragraph (viii) were oxidized to pyridinium dichromate to yield 1 (R)-[3 (S)-( Tert-butoxyformamido) -2-oxo-4-phenylbutyl] -2 (S) -vinylcyclohexane 1.61 g is obtained as a colorless oil which crystallizes; MS m / e 372 [M + H] ⁺ .

(x) In a manner similar to that described in Example 1 (x), 1.44 g of the product of paragraph (ix) was reduced with sodium borohydride to yield 1 (R)-[3 (S)-(tertiary -parts). Oxyformamido) -2 (R) -hydroxy-4-phenylbutyl] -2 (S) -vinylcyclohexane 1.20 g is obtained as a colorless rubber to be crystallized; MS m / e 373 [M] ⁺ .

(xi) In a manner similar to that described in Example 1 (xi), 1.20 g (3.2 mmol) of the product of paragraph (x) were treated with 2,2-dimethoxypropane to give 1 (R)-[[4 ( S)-benzyl-3-(tert-butoxycarbonyl)-2,2-dimethyl-5 (R)-oxazolidinyl] methyl]-2 (S)-1.25 g of vinylcyclohexanes are obtained as a pale yellow rubber do; MS m / e 414 [M + H] ⁺ .

(xii) In a manner similar to that described in Example 1 (xii), 850 mg (2.06 mmol) of the product of paragraph (xi) were treated with potassium permanganate to give 2 (R)-[[4 (S) -benzyl-3 (Tert-butoxycarbonyl) -2,2-dimethyl-5 (R) -oxazolidinyl] methyl] -1 (R) cyclohexanecarboxylic acid 185 mg is obtained as a colorless rubber; MS m / e 432 [M + H] ⁺ .

(xiii) In a manner similar to that described in Example 1 (xiii), 165 mg (0.38 mmol) of the product of paragraph (xii) were coupled with tert-butyl amine to give 2 (R)-[[4 (S) -Benzyl-3-(tert-butoxycarbonyl)-2,2-dimethyl-5 (R)-oxazolidinyl] methyl]-N-tertiary-butyl-1 (R)-cyclohexanecarboxamide 85 mg is obtained as a white solid; MS me / 487 [M + H] ⁺ .

Example 4

2 (R)-[3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tertiary-butyl-1 in a similar manner as described in the first paragraph of Example 2 From (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) -cyclohexanecarboxamide with a melting point of 166.5 to 168 Obtained as a phosphorus white solid; MS m / e 617 [M + Na] ⁺ and 595 [M + H] ⁺ .

2 (R)-[3 (S) -amino-2 (R) -hydroxy-4-phenylbutyl] -N-tertiary-butyl-1 (R) -cyclohexanecarboxamide used as starting material 2 (R)-[3 (S)-(tert-butoxyformamido) -2 (R) -hydroxy-4-phenylbutyl] in a similar manner as described in the second paragraph of Example 2] N-tert-butyl- 1 (R) -cyclohexanecarboxamide (prepared by the method described in the first paragraph of Example 3) and used without further purification.

Example 5

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

2 (S)-[3 (S)-[(L-asparaginyl) amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tertiary-butyl-1 used as starting material R)-cyclohexanecarboxamide is prepared as follows:

2 (S)-[3 (S)-[[N- (benzyloxycarbonyl) -L-asparaginyl] amino] in 10 ml of ethanol-2 (R) -hydroxy-4-phenylbutyl] -N- Tert-Butyl-1 (R) -cyclohexanecarboxamide (prepared by the method described in the first paragraph of Example 2) A solution of 80 mg (0.13 mmol) of hydrogen over 10 mg of 10% palladium on carbon for 4 hours Process. Filtration removes the catalyst and the filtrate is evaporated. The residue is suspended in toluene and then the mixture is evaporated. The process was repeated once to obtain 2 (S)-[3 (S)-[(L-asparaginyl) amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tertiary-butyl- 65 mg of 1 (R) -cyclohexanecarboxamide were obtained as a colorless foam and used without further purification.

Example 6

90 mg of 2 (S)-[3 (S) -amino-2 (R) -hydroxy-4-phenylfutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide in 4 ml of dichloromethane 0.26 mmol) solution of 65 mg (0.26 mmol) of N-benzyloxycarbonyl-cyano-L-alanine in 35 ml of dimethylformamide, 35 mg of hydroxy benzotriazole hydrate (0.23 mmol) and 54 mg of dicyclohexylcarbodiimide ( 0.26 mmol). The mixture is stirred at room temperature under nitrogen for 18 hours and then filtered. The filtrate is evaporated and the residue is partitioned between 25 ml of ethyl acetate and 10 ml of saturated aqueous sodium hydrogen carbonate solution. The organic layer is dried over anhydrous magnesium sulfate and evaporated. The crude product is chromatographed on silica gel column with 3% methanol in dichloromethane as eluent and the product is purified again by crystallization from a mixture of 1 ml dichloromethane and 10 ml hexane. 2 (S)-[3 (S)-[[N- (benzyloxycarbonyl) -3-cyano-L-alanyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N 68 mg of tert-butyl-1 (R) cyclohexanecarboxamide is obtained as a white solid; MS m / e 577 [M + H] ⁺ .

Example 7

In a similar manner to that described in Example 6, 73 mg (0.29 mmol) of N-benzyloxycarbonyl-L-valine was added to 2 (S)-[3 (S) -amino-2 (R) -hydroxy-4 -Phenylbutyl]-N-tertiary-Butyl-1 (R)-Cyclohexanecarboxamide coupling with 100 mg (0.29 mmol) 2 (S)-[3 (S)-[[N-(benzyloxycarbo Nil) -L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide 80 mg is obtained as a white solid; MS m / e 580 [M + H] ⁺ .

Example 8

In a manner similar to that described in Example 6, 78 mg (0.26 mmol) of N-benzyloxycarbonyl-L-phenylalanine was added to 2 (S)-[3 (S) -amino-2 (R) -hydroxy- 4-Phenylbutyl] -N-tert-Butyl-1 (R) -cyclohexanecarboxamide coupled with 90 mg (0.26 mmol) to give 2 (S)-[3 (S)-[[N-(benzyloxy Carbonyl) -L-phenylalanine] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide 72 mg is obtained as a white solid; MS m / e 628 [M + H] ⁺ .

Example 9

In a manner similar to that described in Example 5, 2 (S)-[3 (S)-[[N-(benzyloxycarbonyl)-L-valely]-amino-2 (R)-hydroxy-4 Phenylbutyl] -N-tert-butyl- (1) (R)-95 mg (0.16 mmol) of cyclohexanecarboxamides were hydrotreated on 10% palladium on carbon, and then the product was quinadic acid N-hydroxysuccinimide. Reacted with an ester to give 2 (S)-[3 (S)-[[N- (2-quinolylcarbonyl) -L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N Tert-butyl- (1) R-cyclohexanecarboxamide 50 mg is obtained as a white solid; MS m / e 601 [M + H] ⁺ .

Example 10

In a manner similar to that described in Example 6, 70 mg (0.26 mmol) of N-benzyloxycarbonyl-S-methyl-L-cysteine was added to 2 (S)-[3 (S) -amino-2 (R)- Hydroxy-4-phenylphenyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide coupled to 90 mg (0.26 mmol) to give 2 (S)-[3 (S)-[[N- (Benzyloxycarbonyl) -S-methyl-L-cysteinyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tertiary-butyl-1 (R) -cyclohexanecarbox 30mg Amide Melting Point 143-144 Obtained as a phosphorus white solid; MS m / e 598 [M + H] ⁺ .

Example 11

220 mg (0.59 mmol) of pyridinium dichromate was added to 2 (S)-[3 (S)-[[N- (benzyloxycarbonyl) -L-valyl] amino] -2 (R) -hydride in 1 ml of dimethylformamide. Hydroxy-4-phenylphenyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide 48 mg (0.08 mmol) was added to the solution, and then the mixture was stirred at room temperature for 18 hours. 10 ml of water are added and the mixture is extracted twice with 10 ml fractions of ethyl acetate. The combined extracts are washed twice with 10 ml fractions of water and dried over anhydrous magnesium sulfate and then evaporated. The crude product was chromatographed on silica gel column with 4% methanol in dichloromethane as eluent to give 2 (S)-[3 (S)-[[N-(benzyloxycarbonyl)-L -valyl] amino]- 2-oxo-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide 31mg, melting point 186 to 188 Obtained as a phosphorus white solid; MS m / e 578 [M + H] ⁺ .

Example 12

In a manner similar to that described in Example 6, 72 mg (0.26 mmol) of N-benzyloxycarbonyl-L-aspartic acid β-methyl ester were added to 2 (S)-[3 (S) -amino-2 (R). )-Hydroxy-4-phenylbutyl]-N-tertiary-butyl-1 (R)-cyclohexanecarboxamide with 90 mg (0.26 mmol) for coupling 2 (S)-[3 (S)-[[ N- (benzyloxycarbonyl) -O-methyl-L-aspartyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tertiary-butyl-1 (R) -cyclohexaneca 90 mg of voxamide is obtained as white rubber; MS m / e 610 [M + H] ⁺ .

Example 13

2 (S)-[3 (S)-[(L-asparaginyl) amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 in 2 ml of dimethylformamide R)-60 mg (0.13 mmol) of cyclohexanecarboxamide, 3,5-25 mg (0.13 mmol) of dichlorobenzoic acid, 18 mg (0.13 mmol) of hydroxybenzotriazole hydrate and 27 mg (0.13 mmol) of dicyclohexylcarbodiimide The mixture is stirred at room temperature under nitrogen for 20 hours. The mixture is then filtered and the solid is washed with dichloromethane. The combined filtrates and washings are evaporated to dryness and the residue is partitioned between 50 ml of ethyl acetate and 10 ml of saturated aqueous sodium bicarbonate solution. The separated organic phase is dried over anhydrous magnesium sulfate and then evaporated. The residue is chromatographed on silica gel using 5% methanol in dichloromethane as eluent. The product was further purified by recrystallization from a mixture of 2 ml of dichloromethane and 10 ml of n-hexane to give 2 (S)-[3 (S)-[[N-(3,5-dichlorobenzoyl)-L-asparaginyl] amino ] -2 (R) -hydroxy-4-phenylbutyl] -N-tertiary-butyl- 1 (R) -cyclohexanecarboxamide 37 mg melting point 220-226 Obtained as a phosphorus white solid.

Example 14

In a similar manner to that described in Example 6, 68 mg (0.26 mmol) of N-benzyloxycarbonyl-3-methyl-L-valine was added to 2 (S)-[3 (S) -amino-2 (R)- Hydroxy-4-phenylphenyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide coupled to 90 mg (0.26 mmol) to give 2 (S)-[3 (S)-[[N- (Benzyloxycarbonyl) -3 -methyl-L-valyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-tert-butyl-1 (R) -cyclohexanecarboxamide 40 mg 90 melting point Obtained as a white solid that is (decomposed); MS m / e 594 [M + H] ⁺ .

Example 15

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

The following examples illustrate representative pharmaceutical formulations containing a compound of formula (I) or a pharmaceutically acceptable acid addition salt of a compound of basic formula (I) as the active ingredient.

Example A

The aqueous solution of the active ingredient was sterile filtered, and then in an amount such that 1 ml of the resulting sterile gelatin solution containing phenol as a preservative contained 3 mg of active ingredient, 15 mg of gelatin, 4.7 mg of phenol and distilled water such that the total volume was 1 ml. Mix while using. The mixture is filled into 1 ml vials under sterile conditions.

Claims (8)

Pharmaceutically acceptable acid addition salts of compounds of general formula (I) and compounds of general formula (I) which are basic. Wherein R ¹ is alkoxycarbonyl, araloxycarbonyl, alkanoyl, aralkanoyl, heterocyclylcarbonyl or a group of formula (i), Wherein R ⁸ is alkoxycarbonyl, aralkoxycarbonyl, alkanoyl, aroyl, aralkanoyl or heterocyclylcarbonyl, and R ⁹ is alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, cyanoalkyl , Carbamoylalkyl, alkylthioalkyl, alkoxyalkyl or alkoxycarbonylalkyl), R ² is alkyl, cycloalkylalkyl or aralkyl, R ³ is hydrogen, R ⁴ represents hydroxy, or R ³ And R ⁴ together represent oxo, R ⁵ is alkoxycarbonyl or alkylcarbamoyl, and R ⁶ and R ⁷ together are trimethylene or tetramethylene unsubstituted or substituted by tetramethylene on alkyl or on adjacent carbon atoms. Indicates. The compound of claim 1, wherein R 1 is tert-butoxycarbonyl or a group of formula wherein R ⁸ is benzyloxycarbonyl, 3,5-dichlorobenzoyl or 2-quinolylcarbonyl, and R is ⁹ is isopropyl, tert-butyl, benzyl, cyanomethyl, carbamoylmethyl, methylthiomethyl or methoxycarbonylmethyl), R ² is benzyl, R ³ is hydrogen, R ⁴ is hydroxy And R ⁵ is methoxycarbonyl or tert-butyl carbamoyl, and R ⁶ and R ⁷ together represent unsubstituted tetramethylene. 2 (S)-[3 (S)-[[N- (2-quinolylcarbonyl) -L-asparaginyl] amino] -2 (R) -hydroxy-4-phenylbutyl] -N-3 Class-Butyl-1 (R)-Cyclohexane-Carboxamide. (a) A compound of formula (II), characterized by treating with an acid, wherein R ¹ is alkoxycarbonyl or alkoxycarbonyl, R ³ is hydrogen and R ⁴ is hydroxy. A compound of formula (I) or a pharmaceutically acceptable salt thereof in the form of an optically pure diastereomer, diastereomeric mixture, diastereomeric racemate or diastereomeric racemate mixture according to any one of How to manufacture. Wherein R ^1a is alkoxycarbonyl or aralkoxycarbonyl, and R ² , R ⁵ to R ⁷ are as defined in claim 1. A medicament for preventing and treating a viral infection, comprising the amino acid derivative according to any one of claims 1 to 3 and a therapeutically inactive excipient. A compound of general formula (III) is reacted with an acylating agent which introduces a group R ¹ as defined in claim 1, wherein the optically pure diastereomer according to any one of claims 1 to 3. , A process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof in the form of a diastereomeric mixture, a diastereomeric racemate or a diastereomeric racemate mixture. Wherein R ¹ to R ⁷ are as defined in claim 1. A compound of formula (IV) is reacted with an acylating agent which introduces a group R ⁸ as defined in claim ¹ , wherein R ¹ is a group of formula (i). A compound of formula (I) or a pharmaceutically acceptable salt thereof in the form of an optically pure diastereomer, diastereomeric mixture, diastereomeric racemate or diastereomeric racemate mixture according to any one of How to manufacture. Wherein R ² to R ⁹ are as defined in claim 1. The compound according to any one of claims 1 to ³ , characterized in that the compound of formula (I) wherein R ³ is hydrogen and R ⁴ is hydroxy, wherein R ³ and R ⁴ together represent oxo. A process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof in the form of an optically pure diastereomer, diastereomeric mixture, diastereomeric racemate or diastereomeric racemate mixture. Wherein R ¹ , R ² and R ⁵ to R ⁷ are as defined in claim 1.