KR960000078B1 - Hiv protease inhibitors and process for the preparation thereof - Google Patents

Abstract

The protease inhibitors (I) of A-B-NH-CHR1-epoxy-CH2-CO-D-E consist of (1) coupling cis-epoxy deriv. as PNH-CHR1-epoxy-CH2-COOH (II) with H-D-E, (2) coupling with A-B-OH. R1 is cycloalkyl or arylalkyl, A is aryloxyalkylcarbonyl or arylalkoxycarbonyl, B and D are amino acid, E is lower alkoxy, lower alkylamine, aryl or cycloalkylamine. The chemical composition and NMR and Mass spectrum data of new 24 compounds of (I) are described. The IC50 of (I) is 0.2-10.0 micromole, KI is 0.018-1.057 micromole, and Kina is 0.10-0.43 per minute.

Description

Inhibitory Compounds of Human Immunodeficiency Virus Protease and Methods for Making the Same

The present invention relates to novel compounds that inhibit human immunodeficiency virus (HIV) proteases, methods for their preparation, and compositions for the treatment or prevention of epistatic immune deficiency syndrome (AIDS) caused by HIV infection.

HIV is a retrovirus that retains genetic information in the form of RNA. The virus's structure consists of RNA, the innermost genetic information, and a reversetranscriptase that can reverse DNA from the RNA, which is surrounded by a shell protein, and on the outside is a lipid membrane. This protective film is formed. The lipid membrane is embedded with glycoproteins called gp-120 and gp-40, which are known to play a critical role in the virus' recognition and penetration of T-cells.

The more complex the mechanism of action of a virus, the more likely it is to develop a method of combating it. Since HIV has the most complex form of replication among many viruses, various forms of treatment have been developed. The best known therapeutic agents to date are reverse transcriptase inhibitors. Reverse transcriptase is an enzyme that replicates DNA from RNA. Because of this specificity, it is an enzyme that exists only in RNA viruses, ie retroviruses. Because of their presence only in retroviruses, it was expected that compounds that inhibit these reverse transcriptases would be able to neutralize HIV with minimal side effects. Accordingly, many reverse transcriptase inhibitors have been developed. Azidothymidine (AZT) from Boro-Welcome, 2 ', 3'-dideoxyinosine (DDI) from Bristol-Myers Squibb, and 2', 3'-dideoxytocin (DDC) from Hoffman La Roche This is the case. However, most of these compounds currently used have only a life-extending effect rather than a complete cure, and cause side effects such as decreased platelet count and decreased bone marrow cells.

Another important therapeutic agent is an HIV protease inhibitor. HIV does not synthesize each protein from mRNA when it makes a skin protein and the necessary enzymes, but gag-protein (P55) or gag-, a long form of proprotein that combines both skin proteins and enzymes together. The pol protein (P165) is made first, and then the protein is broken down by the protease in it into enzymes necessary for the formation of virus such as shell protein, reverse transcriptase and integrase. Inhibiting the protease responsible for this degradation process stops the replication of the virus, and protease inhibitors are based on this principle. The HIV protease consists of 99 amino acids, two identical units exist as dimers, and the molecular weight of each unit value is 10793. HIV proteases are aspartic proteases that have a typical configuration of aspartate-threonine-glycine at the reaction site. The development of HIV protease inhibitors follows the trend of developing inhibitors of other types of aspartic proteases (particularly Lenin).

The basic approach is to develop a compound that is similar to the enzyme's transition state (TSA) to increase its affinity for the enzyme, thereby enhancing its binding to the enzyme. HIV protease inhibitors developed in this manner are disclosed in several documents (Roberts, at al., Scinence 248, 358 (1990); European Patent Application Publications 0337714; 0346847; 356223; 352000; 357332; 357332; 362002; and 361341; Korean Patent Publication Nos. 90-18134; bone et al., JACS 113, 9382 (1991).

However, all of these known HIV protease inhibitors have the incidence of reversible inhibitors with increased affinity for enzymes.

Accordingly, the present inventors have conducted research to develop irreversible inhibitors that can solve the above-mentioned problems. As a result, the inventors have developed a new compound containing cis-epoxide as described below.

In an attempt to suppress HIV protease using epoxides, Moeling et al. Reported using a natural product, serurenin (Moeling, et al., FEBS Letter 261, 373 (1990); Pal, et al., PNAS 85, 9283 (1988)), its inhibitory effect is so low that the potential for therapeutic use is questioned. Therefore, neither has been described or mentioned for the use of cis-epoxides as cited in the present invention as irreversible inhibitors or as anti AIDS agents.

Hereinafter, the present invention will be described in detail.

The present invention provides a cis-epoxide derivative represented by the following general formula (I) and a pharmaceutically acceptable salt thereof.

In the above formulas, R ¹ is a cycloalkyl or lower alkyl group substituted with aryl; A is a aryloxyalkylcarbonyl group, an aryl alkoxycarbonyl group or a carbonyl group substituted with an aromatic heterocyclic system containing a nitrogen atom; B is an amino acid residue; D is an amino acid residue or a residue in which two amino acids are the same or different from each other; E is a lower alkoxy group, a lower alkylamine group, a lower alkaliamine group substituted with an aromatic heterocyclic ring containing aryl or a nitrogen atom, or a cycloalkylamine group fused with an aromatic ring and substituted with hydroxy.

As used herein, the term "lower alkyl" refers to straight or branched chain alkyl of 1 to 4 carbon atoms, including methyl, isopropyl, isobutyl, t-butyl.

More preferred among the compounds of the present invention as defined above include those in which the general formula (I) R ¹ is cyclohexyl methyl or benzyl, and A is phenoxymethylcarbonyl, naphthoxymethylcarbonyl, quinalinylcarbon N, pyrimidinylcarbonyl, benzimidazolylcarbonyl or benzylcarbonyl, B is an asparagine or valine residue, D is an isoleucine, phenyl alanine, glutamine, or valine residue or any two of these amino acids Is bonded, E is methoxy, ethoxy, methylamino, dimethylamino, ethylamino, t-butylamino, benzylamino, benzylamino, benzimidazol-2-yl-methyleneamino, pyridin-2-yl- Included are compounds that are methyleneamino or (2R) -hydroxy- (1S) -indanyl amino groups.

Other compounds in the present invention may also have asymmetric carbon centers and exist as racemates, racemic compounds, diastereomeric mixtures and individual diastereomeric stereoisomers, all of which isomeric forms are included in the present invention.

The main compounds according to the invention are as shown in Table 1 below.

TABLE 1

.

.

The compounds of formula (I) according to the invention can be prepared by methods I, II and III.

Method I

In the above formulas, R ¹ , A, B, D and E are as defined above; P is a protecting group, preferably benzyloxycarbonyl or t-butoxy carbonyl group.

The method I is a method of using an epoxide compound as a starting material, the compound of the general formula (IV) by coupling the epoxide compound of the general formula (II) with the compound represented by the general formula (III) Coupling reaction with to obtain the desired compound of general formula (I).

Method II

In the above formulas, R ¹ , A, B, D and E are as defined above; P is a protecting group, preferably benzyloxycarbonyl or t-butoxy carbonyl group.

According to the method II, the compound of the general formula (VIII) is reacted by coupling the compound of the general formula (VI) with the compound of the general formula (III) to epoxidize the compound of the general formula (VII). The protective group is removed from the compound of the general formula (VIII) and then subjected to a coupling reaction with the compound of the general formula (V) to obtain the compound of the general formula (I).

Method III

In the above formulas, R ¹ , A, B, D and E are as defined above; P is a protecting group, Preferably it is a benzyloxycarbonyl group or t-butoxy carbonyl group.

According to the above method III, the compound of the general formula (VI) is reacted with the compound of the general formula (III) to obtain the compound of the general formula (VII), and the protecting group from the compound of the general formula (VII) After removal, the compound is reacted with the compound of the general formula (V) to obtain the compound of the general formula (IX), and then the compound of the general formula (IX) is subjected to an epoxidation reaction to obtain the compound of the general formula (I). Obtain the compound.

Coupling reactions in methods I, II and III can be carried out according to methods commonly used in the art. For example, dicyclohexylcarbodiimide (DCC), 3-ethyl-3 '-(dimethylamino) -propylcarbodiimide (EDC), bis- (2-oxo-3-oxazolidinyl) -phosphinic acid The reaction can be carried out in the presence of a coupling reagent such as chloride (BOP-Cl), diphenylphosphoryl azide (DPPA) and the like. Such coupling reagents need not be limited to those listed above. Alternatively, the reaction may be carried out after the carboxylic acid of Formula (II), (V) or (VI) is made of an acid halide or other active ester without a coupling reagent. Acid chloride may be easily used as the acid halide, and active esters are those commonly used to activate a carboxylic acid group, and the carboxylic acid compound may be, for example, an alkoxycarbon such as methoxycarbonyl chloride, isobutoxycarbonyl chloride, or the like. Nylhydride or N-hydroxybenzotriazole, N-hydroxyphthalimide, N-hydroxysuccinimide, N-hydroxy-5-norbornene-2,3-dicarboxamide, 2,4, Esters derived by reacting with carboxylic anhydrides derived from coupling reagents such as 5-trichlorophenol and the like.

On the other hand, the epoxidation reaction in the above method II and III is carried out according to the method commonly used in this field.

The protecting group leaving reaction may also be carried out by a conventional method according to the type of protecting group. For example, when the protecting group is benzyloxycarbonyl, the protecting group may be removed by reaction under hydrogen pressure in the presence of a Pd / C catalyst, and the protecting group may be t-part. In the case of oxycarbonyl, it can be removed by reacting with trifluoroacetic acid.

As described above, the compounds of formula (III) and compounds of formula (VI) used as starting materials in the preparation methods I, II and III of the present invention may be prepared according to the methods described below as novel substances. Can be.

First, the compound of general formula (II) is prepared by the following reactions.

In the above formulas, R1 and P are as defined above, 9 '' is a protecting group and X is Cl, Br or I as the halogen atom.

First, the alcohol site of 3-halo-propan-1-ol is protected and then reacted with triphenylphosphine to obtain a phosphonium salt of the general formula (X), and the compound of the general formula (X) is And react with an aldehyde of XI) to obtain a compound of formula (XII). Then, the protecting group P '' is removed from the compound of the general formula (XII) and then subjected to an epoxidation reaction in the same manner as described above to obtain a compound of the general formula (XIII); Subsequently, the alcohol site | part of this compound of general formula (XIII) is oxidized, and the compound of the general formula (II) can be obtained.

In the above formulas, R 'and P are as defined above and X is Br or I as halogen atom.

This is based on the method described in Keinan et al., Tetrahedron 47, 4631-4638 (1991) and Corey & Shimaji, JACS 105, 1662-1664 (1983) as a process for preparing cis-β, γ-olefin acids. It is a process. In the case of synthesizing the compound of the general formula (VI) by acid treatment of the ortho ester of the general formula (XV) according to the above paper to form a compound of the general formula (XVI) and then base treatment to remove the boro groups 50 More than% is converted from [beta]-[gamma] olefinic acid to [alpha]-[beta] olefinic acid so that the yield of the desired compound is very low and the separation thereof is difficult. Accordingly, the present invention relates to the general formula (VI) which is not converted to α-β olefinic acid by heating and stirring the orthoester of the general formula (XV) compound with t-butanol in the presence of an acid catalyst as a solvent to the reflux temperature of the solvent. Compounds can be obtained in 80-90% yield.

The compounds of the present invention are useful for the treatment or prevention of AIDS or HIV infection.

Compounds of the present invention may be administered at a total daily dose of 0.01-10 mg / kg body weight to be given to the host, either once or several times, and specific dose levels for a particular patient may be determined by the specific compound, weight, age, May vary depending on sex, health, diet, time of administration, method of administration, rate of excretion, drug mixing and severity of disease.

The compounds of the present invention can be administered in injectable and oral formulations as desired.

Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be prepared using suitable dispersing agents, wetting agents or suspending agents according to known techniques. Acceptable solvents that can be used include water, Ringer's solution and isotonic NaCl solution, and sterile fixed oils are conventionally employed as a solvent or suspending medium.

Any non-irritating fixed oil may be used for this purpose, including mono- and diglycerides, and fatty acids such as oleic acid are used as injectables.

In the case of a solid dosage form for administration, capsules and tablets are possible, but in view of compound properties, the capsules are particularly preferable, and in the case of tablets, it is useful to prepare them as enteric agents. The solid dosage form may contain one or more inert diluents, such as sucrose, lactose or starch, and a lubricant such as magnesium squarate in addition to the diluent.

The compounds of the present invention may be administered in combination with one or more other anti-AIDS agents, immunomodulators, and the like.

Such anti-AIDS agents that can be administered in combination with a compound of the present invention include zidovudine (AZT), dididioxyinosine (DDI), dididioxycycin (DDC), ribavirin, compound Q, alpha interferon, beta interferon, polymanno Acetate, sodium, phosphonoformate, HAD-23, UA001, eflornithine, acyclovir.

Immunomodulators that can be administered in combination with a compound of the invention include bropyrimin, amprigen, anti-human alpha interferon antibody cloni stimulator, CL246738, Imp 2-2, diethyldithiocarbamate, interleukin-2 , Alpha-interferon, inosinepranovex, methionine enkephalin, muramil-tripeptide, TP-5, erythropoietin, naltrexone and payload lysine.

Formulations of the compounds of the present invention for the treatment and prevention of HIV infection are not limited to those described above, and any flexible formulation of the compounds of the present invention necessary for the treatment and prevention of HIV infection may be included.

This invention will be described in more detail based on the examples. The following examples are only intended to facilitate understanding of the preparation method of the novel compounds according to the present invention, but do not limit the scope of the present invention.

Example 1

Step 1:

5-L- (N-benzyloxycarbonyl) amino-6-phenyl-hex-3- (cis) -enyl-4'-methyl-2 ', 6', 7'-trioxa-bicyclo [2 ' , 2 ', 2'] Synthesis of oxetane

4.87 g (10 mmol) synthesized according to Keinan et al; Tetrahedran 26, 4631-4638 (1991) was dissolved in 50 ml of 1,4-dioxane and 5.1 g (30 mmol) of potassium carbonate in powder form. 2.5 g (10 mmol) of L- (N-benzyloxycarbonyl) phenylalanal was added thereto, followed by stirring for 15 hours at the reaction temperature of the reaction solvent After cooling, the reaction solvent was diluted with 50 mg of ethyl acetate. The organic layer was dried over anhydrous MaSO ₄ to remove the solvent, and then subjected to column chromatography (hexane: ethyl acetate = 7: 3) to obtain 2.5 g (60% yield) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.8 (s, 3H), 2.2 ~ 3.0 (m, 4H), 3.9 (s, 6H), 4.6 (m, 1H), 4.8 (br, 1H), 5.05 (s, 2H ), 5.4 to 5.6 (m, 2H), 7.1 to 7.5 (m, 10H)

Step 2:

Synthesis of 5-L- (N-benzyloxycarbonyl) amino-6-phenyl-hex-3- (cis) -ene-1-carbonyl acid

2.5 mg (6 mmol) of the compound obtained in step 1 was dissolved in t-butane octane containing 10% HCl, stirred at reflux of the reaction solvent for about 20 hours, the reaction solvent was distilled off under reduced pressure, and diluted with ethyl acetate. After washing with K ₂ CO ₃ solution. The organic layer was removed, the water layer was adjusted to pH 2, extracted with ethyl acetate, treated with anhydrous MaSO ₄ , and the organic solvent was removed to obtain 1.62 g (80% yield) of the title compound.

¹ H NMR (CDCl ₃ ) δ 2.7 ~ 3.3 (s, 4H), 4.6 (m, 1H), 4.8 (br, 1H), 5.05 (s, 2H), 5.4 (t, 1H), 5.6 (m, 1H ), 7.1 to 7.3 (m, 10H)

Mass (FAB, m / e) 340 (M + 1)

Step 3:

Synthesis of N- [5-L- (N-benzyloxycarbonyl) amino-6-phenyl-hex-3- (cis) -ene-1-carbonyl] -isoleucine methylester

339 mg (1 mmol) of the compound obtained in Step 2 was dissolved in anhydrous dichloromethane, and 1 equivalent of isoleucine methyl ester and 4 steps of triethylamine were added, followed by cooling to 0 ° C. 1.5 equivalents of POCl ₃ was slowly added dropwise to the solution, diluted with dichloromethanol after 3 hours, washed with basic water, dried over anhydrous MaSO ₄ , and the solvent was distilled off under reduced pressure. Then, column chromatography (hexane: ethyl ester = 7: 3) was carried out to obtain 256 mg (55% yield) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.9 (m, 6H), 1.1 ~ 1.4 (m, 2H), 1.8 (m, 1H), 2.6 ~ 2.9 (m, 3H), 3.2 (m, 1H), 3.7 (s , 3H), 4.4 ~ 4.6 (m, 2H), 4.8 ~ 5.1 (m, q, 3H), 5.4 (d, 1H), 5.6 (m, 1H), 6.8 (m, 1H), 7.1 ~ 7.4 (m , 10H)

Step 4:

Synthesis of N- [5-L- (N-benzyloxycarbonyl)-(4R, 3S) -epoxy-6-phenyl-hexanoyl] -L-isoleucine methyl ester

256 mg (5.5 mmol) of the compound obtained in step 3 was dissolved in anhydrous dichloromethane, and then 2 equivalents of 50% metachloroperbenzoic acid (mCPBA) was added and stirred for about 20 hours. After a certain time, the organic layer was washed with 10% NaS ₂ HCO ₃ solvent. The organic layer was dried over anhydrous MgSO ₄ , and then the organic solvent was distilled off under reduced pressure, followed by column chromatography (hexane: ethyl acetate = 1: 1) to obtain 212 mg (80% yield) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.9 (m, 6H), 1.1 ~ 1.4 (m, 2H), 1.8 ~ 2.3 (m, 3H), 2.7 ~ 3.4 (m, 4H), 3.7 (s, m, 4H) , 4.6 (m, 1H), 5.1 (br, s, 3H), 6.3 (br, 1H), 7.1 ~ 7.5 (m, 10H)

Mass (FAB, m / e) 483 (M + 1)

Step 5:

Synthesis of Methyl Ester (Compound 2 in Table 1)

212 mg (0.44 mmol) of the compound obtained in step 4 was dissolved in anhydrous methane jade, 10% Pd / C was added 40 mg, and stirred for about 5 hours under H ₂ condition (rubber balloon) to remove carbobenzoyloxy, and anhydride was added to celite. After filtration and removal of the organic solvent, 1 equivalent of N- (2-quinolinecarbonyl) -L-asparagine, 1 equivalent of EDC, 1.2 equivalents of HOBT, and 1.5 equivalents of triethylamine were dissolved in anhydrous dimethylformamide (3 ml) for about 20 hours. Stirred at ambient temperature. Dimethylformamide was removed by distillation under reduced pressure, diluted with dichloromethane solvent, and washed with saturated NaHCO ₃ solvent. After drying the organic layer with anhydrous MgSO ₄ , column chromatography (EtOAc = 100%) was carried out to give 190 mg (70% yield) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.9 (m, 6H), 1.2 ~ 1.5 (m, 2H), 1.9 (m, 1H), 2.3 ~ 3.4 (m, 6H), 3.7 (s, 3H), 4.2 ~ 4.6 (m, 2H), 5.05 (m, 1H), 5.8 to 6.6 (s, s, 2H), 6.9 to 7.5 (m, 6H), 7.6 to 8.4 (m, 6H), 8.9 to 9.4 (d, 1H)

Mass (FAB, m / e) 618 (M + 1)

Example 2

Step 1:

Synthesis of 2- [5-L- (N-benzyloxycarbonyl) amino) -6-phenyl-hex-3- (cis) -en-1-oxy] -tetrahydropyran

4.5 g (11 mmol) of N-benzyloxycarbonyl-phenylalanal and 7.5 g (16.5 mmol) of 2- (3-triphenylphosphonium-propane-1-oxy) -tetrahydropyran iodide salt It was dissolved in 150 ml of 1,4-dioxane with g (33 mmol) of K ₂ CO ₃ and refluxed for 20 hours. The reaction mixture was cooled, diluted with 50 ml of ethyl acetate, and washed with water. The organic layer was dried over anhydrous MgSO ₄ , the solvent was removed, and purified by column chromatography (hexane: ethyl acetate = 8: 2) to obtain 4.2 g (93% yield) of the title compound. This product contained cis-olefin: trans-olefin in a ratio of 6: 1.

¹ H NMR (CDCl ₃ ) δ 1.4 ~ 1.7 (m, 6H), 2.1 (m, 2H), 3.6 ~ 3.9 (m, 2H), 3.1 (m, 1H), 3.4 (m, 1H), 3.5 (m , 1H), 3.7 (m, 1H), 4.4 (s, 1H), 5.0 (s, 2H), 5.2 (m, 1H), 5.4 (m, 1H), 7.3 ~ 7.3 (m, 10H)

Mass (FAB, m / e) 410 (M + 1)

Step: 2

Synthesis of 5-L- (N-benzyloxycarbonyl) amino) -6-phenyl-hex-3- (cis) -en-1-ol

12 g (20 mmol) of the compound obtained in step 1 was dissolved in 450 mg of ethanol, and 1.2 g of pyridinium-para-toluenesulfonate (PPTS) was added thereto, followed by heating at 70 ° C. for 6 hours. The solvent was distilled off under reduced pressure, and column chromatography (hexane: ethyl acetate = 8: 2) gave 7.3 g (75% yield) of the title cis-olefin and 1.2 g (12%) of trans-olefin.

¹ H NMR (CDCl ₃ ) δ 2.0 ~ 2.4 (m, 2H), 3.6 ~ 3.9 (m, 2H), 3.5 (m, 2H), 4.5 (m, 1H), 5.0 (m, s, 3H), 5.3 5.5 (m, 2H), 7.1-7.4 (m, 10H)

Step 3:

Synthesis of 5-L- (N-benzyloxycarbonyl) amino) -6-phenyl- (4R, 3S) -epoxy-hexane-1-1ol

10 g of metachlorobenzoic acid of 4 g of the compound obtained in Step 2 was dissolved in 300 ml of dichloromethane, and stirred at room temperature for 10 hours. 5 g of Na ₂ S ₂ O ₃ was added thereto, followed by further stirring for 30 minutes, followed by extraction with ethyl acetate and saturated aqueous NaHCO ₃ solution. The organic layer was dried over anhydrous MgSO ₄ and distilled under reduced pressure to obtain 3.89 (90% yield) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.3 ~ 1.6 (m, 2H), 2.8 ~ 3.1 (m, 4H), 3.7 (t, 2H), 3.8 (m, 1H), 5.0 (br, 1H), 5.1 (s , 2H), 7.2 to 7.4 (m, 10H)

Mass (FAB, m / e) 342 (M + 1)

Step 4:

Synthesis of 5-L- (N-benzyloxycarbonylamino) -6-phenyl-hex- (4R, 3S) -epoxy-1-carbonyl acid

324 mg of the compound obtained in step 3 was dissolved in anhydrous DMF, 2.5 g (6.6 mmol) of pyridium dichloromate was added thereto, followed by stirring at room temperature under nitrogen for 100 hours. The reaction mixture was filtered through celite and washed with 150 ml of ethyl acetate. After washing with IM KHSO ₄ aqueous solution, the organic layer was dried over anhydrous MgSO ₄ and distilled under reduced pressure to obtain 280 ml (80% yield) of the title compound.

¹ H NMR (CDCl ₃ ) δ 2.0 (m, 2H), 2.8 ~ 3.1 (m, 3H), 3.3 (m, 1H), 3.6 (m, 1H), 5.1 (s, 2H), 5.9 (d, 1H ), 7.1 to 7.4 (m, 10H)

Mass (FAB, m / e) 356 (M + 1)

Step 5:

Synthesis of N- [5-L- (N-benzyloxycarbonylamino)-(4R, 3S) -epoxy-6-phenyl-hexanoyl] -L-isoleucine methylester

After dissolving the compound obtained in step 4 and L-isoleucine-methyl ester in 1 equivalent of dimethylformamide, the same amount of EDC, HOBT and FABMS results were obtained in the same manner as in step 5 of Example 1, respectively. Same as in step 4.

Step 6:

Synthesis of Methyl Ester (Compound 2 in Table 1)

The title compound was obtained in the same manner as in Step 5 of Example 1, above. NMR and FABMS results were the same as in step 5 of Example 1 above.

[Examples 3 to 25]

Compounds 1 and 3 to 24 listed in Table 1 were synthesized in the same manner as in Example 1 or 2, and their NMR and Mass results are reported in Table 2 below. However, (N-benzyloxycarbonyl) -L-cyclohexylphenyl alanineal was synthesized according to Boger et al., JMC 28, 1779-1790 (1985), and (2R) -hydroxy- (1S)- Amino-indane was prepared according to Thompson et al., JMC 35, 1685-1701 (1992).

TABLE 2

Assay for Screening HIV Protease

The compounds of the present invention used the following method to determine the inhibitory effect on HIV protease.

An oligopeptide consisting of 11 amino acids of Ser-Nle-Ala-Glu- (p-NO ₂ ) -Pha-Leu-Val-Arg-Lys-His was used as the reactor. This substrate was subjected to HIV protease using a strong absorbance at 280 mm of (p-NO ₂ ) -Phe to separate the substrate before reaction and the product after reaction by HPLC to determine the relative amount of product.

Compounds of the present invention are ultimately an irreversible inhibitor having an inhibitory effect of 100%. Table 3 shows the irreversible inhibition constants of certain compounds of the present invention.

TABLE 3

Antibacterial Activity Assay

The anti-HIV activity of the compounds according to the invention was measured by the following method.

1 × 10 ⁵ cells of H9 and Sup T1 were placed in 24 well plates, and various concentrations of the compound of the present invention were added. The HIV-1 inoculum of 50 TCID ₅₀ was added thereto, and in the case of Sup T1, syncytia formation was examined after 3 days. In the case of H9, three quarters of the culture solution was changed to a new culture solution containing the same concentration of inhibitor at three days intervals. After 9 days, 1 ml of the solution was centrifuged, the supernatant was placed in a tube containing 0.9 ml of 0.1 ml 5% Triton X-100, and the virus was neutralized, and then tested for HIV-1P. In case of reverse transcriptase assay, 1 ml of supernatant and 0.5 ml 30% PEG were added, and the virus was allowed to precipitate by centrifugation at 4 ° C. overnight. The precipitate was placed in a buffer solution containing Triton X-100 and subjected to reverse transcriptase assay.

Cell toxicity test to determine the maximum allowance of anti-AIDS agent was carried out by adding compounds ranging from 0.1 uM to 100 uM in H9 cells and Sup T1 cells and changing the culture medium every three days to determine the extent of cell proliferation. Observation was made for 2 weeks by the trypan blue dye exclusion tehnique.

Table 4 below shows the antibacterial activity and cytotoxicity test results of certain compounds of the present invention.

TABLE 4

Claims (7)

Cis-epoxide derivatives represented by the general formula (I), pharmaceutically acceptable nontoxic salts thereof, isomers, hydrates and solvates thereof. In the above formulas, R ¹ is a cycloalkyl or lower alkyl group substituted with aryl; A is a carbonyl group substituted with an aryloxyalkylcarbonyl group, an allyl alkoxycarbonyl group or an aromatic heterocyclic system containing a nitrogen atom containing a nitrogen atom; B is an amino acid residue; D is an amino acid residue or a residue in which two amino acids are the same or different from each other; E is a lower alkoxy group, a lower alkali amine group, a lower alkali amine group substituted with an aromatic heterocyclic ring containing aryl or a nitrogen atom, or a cycloalkylamine group fused with an aromatic ring and substituted with hydroxy. A compound according to claim 1, wherein R ¹ is a cyclohexylmethyl or benzyl group, A is phenoxymethylcarbonyl, naphthoxymethylcarbonyl, quinalylcarbonyl, pyrimidinylcarbonyl, benzimidazolylcarbonyl or Benzylcarbonyl group, B is an asparagine or valine residue, D is an isoleucine, phenylalanine, glutamine or valine residue, or any two amino acids thereof are combined, E is methoxy, ethoxy, methylamino, dimethyl A compound which is an amino, ethylamino, t-butylamino, benzylamino, benzimidazol-2-ylmethyleneamino, pyridin-2-yltyleneamino or (2R) -hydroxy- (1S) -indanylamino group. Cis-epoxide compound represented by the following general formula (II). Wherein R ¹ is as defined in claim 1 above; P is benzyloxycarbonyl or butyloxycarbonyl group. The cisolefin compound represented by the following general formula (VI). Wherein R ¹ is as defined in claim 1 above; P is benzyloxycarbonyl or butyloxycarbonyl. A method for producing a cis olefin compound represented by the following general formula (VI) by heating the reflux product of the following general formula (XV) with t-butanol as a solvent in the presence of an acid catalyst to the reflux temperature of the solvent. In the above formulas, R ¹ and P are as defined in claim 5 above. A composition for the treatment or prophylaxis of a human immunodeficiency virus infection containing a therapeutically effective amount of another compound of claim 1, which contains at least one adjuvant selected from a pharmaceutically acceptable carrier, dispersant, wetting agent, inert diluent, excipient and lubricant. .