KR960000077B1 - Hiv protease inhibitors and process for the preparation thereof - Google Patents

Abstract

The protease inhibitors (I) of amino(3R,4S)-epoxy alkane as A-B-CHR1-NH-epoxy-R2 consist of (1) esterifying phenylalanine derivatives, (2) hydrating (1) with diisobutylaluminum, (3) coupling with A-B-OH, (4) forming epoxide with meta-chloroperoxybenzoic acid. R2 is one of diphenyl hexane, 1-phenyl octane. The chemical composition and NMR and Mass spectrum data of new 23 compounds of (I) are described. The IC50 of (I) is 0.2-5.0 micromole, KI is 0.025-44.4 micromole, and Kina is 0.03-0.45 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 acquired 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 contains RNA, the innermost genetic information, and reverse transcriptase, which can reverse DNA from the RNA, which is surrounded by a shell protein, and outside of it. The lipid film forms a protective film. 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 for 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. Azidothimidine (AZT) from Boro-Welcome, 2 ', 3'-dideoxyinosine (DDI) from Bristol-Myers Squibbs, and 2', 3'-dideoxycytosine (DDC) from Hoffman La Roche ) And the like. However, these compounds are currently only used for their life-extending effects, rather than completely treating the disease, causing 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 bark protein and necessary enzymes, but gag-protein (P55) or gag-pol, which is a long form of protein that is combined with bark protein and enzyme. The protein (P165) is made first, and then the protein is broken down by the proteases 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 present as dimers, and each unit has a molecular weight of 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 resins).

The basic approach is to increase the affinity to the enzyme by developing a compound (transition state analogue, TSA) similar to the enzyme's transition state. HIV protease inhibitors developed in this way have been disclosed in several documents (Roverts, at al., Science 248, 358 (1990); European Patent Application Publications 0337714; 0346847; 356223; 352000; 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 disadvantage that they are 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 present inventors have developed new compounds including 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.

Wherein R ¹ and R ² may be the same or different and each is a lower alkyl group or an arylalkyl group; A is R ^3- (CH ₂ ) ₁ Wherein R ³ is an aryl group, an aryloxy group substituted or unsubstituted with lower alkyl or halogen or hydroxy, an aromatic heterocyclic system containing 1 to 3 nitrogen atoms or an N-oxide system thereof, or oxygen and nitrogen A 3 to 6 membered heterocyclic system containing atoms at the same time or containing only nitrogen atoms, 1 being 0, 1 or 2), Wherein R ⁴ is a lower alkyl group or an arylalkyl group, or ; Wherein R ⁵ is a substituted group boson ring system comprising oxygen and nitrogen atoms; B is group represented by following General formula (a).

Wherein R ⁴ is a lower alkyl group or a carbamoylalkyl group, R 7 is a lower alkyl group or an arylalkyl group, and m and n are each 0 or 1.

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, especially when lower alkyl is a bridge It is a linear alkyl of.

As used herein, "aryl" refers to a monovalent mono or dicyclic aromatic hydrocarbon group such as phenyl, naphthalene and the like.

In the present specification, the "aromatic heterocyclic system containing 1 to 3 nitrogen atoms" refers to a mono or cyclic aromatic heterocyclic system including 1 to 3 nitrogen atoms in a ring such as quinoline or quinoxalinyl, and The N-oxide system ˝ refers to a form in which an oxygen atom is bonded to a nitrogen atom inside the aromatic heterocycle.

In the present specification, the 3- to 6-membered heterocyclic system including oxygen and nitrogen atoms or only nitrogen atoms includes at least one nitrogen atom or nitrogen atom in the ring, such as imidazole, triazole, and morpholine. And an alicyclic or aromatic heterocyclic system including oxygen atoms at the same time.

Among the compounds of the present invention as defined above, a more preferable compound is, in the general formula (I), R ¹ is an isobutyl group or a benzyl group; R ² is a 2-phenylethyl group or an isopentyl group; A is R ^3- (CH ₂ ) ₁ Wherein R ³ is phenoxymethyl, 2,4-difluorophenoxymethyl, naphthoxymethyl, quinolinyl, oxyquinolinyl, morpholinyl, morpholinpropyl or quinoxalinyl, 1 is 0 , One or two, And R ⁴ is t-butyl or benzyl, or It is represented by and R <^5> is group which is morpholine; B are those in which asparagine, alanine, phenylalanine, glutamine or valine residues are alone or optionally two linked residues.

The compounds according to the invention may also have an asymmetric carbon center and may exist as racemates, racemic compounds, diastereomeric mixtures and individual diastereoisomers, all of these isomeric forms being included in the invention.

Representative compounds according to the invention include 2-L- (t-butoxycarbonyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-benzyloxycarbonyl-L -Asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-benzyloxycarbonyl-L-valinyl) amino- (3R, 4S) -epoxy- 1,6-diphenylhexane, 2-L- (N-benzyloxycarbonyl-L-isorosinyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- ( N-benzyloxycarbonyl-L-glutaminyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 5-L- (N-benzyloxycarbonyl-L-asparaginyl) amino -(3R, 4S) -Epoxy-7-methyl-1-phenyloctane, 2-L- (N-phenoxyacetyl-L-asparaginyl) amino (3R, 4S) -epoxy-1,6-diphenyl Hexane, 2-L- (N-1-naphthoxyacetyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-2-quinolinecarbo Nyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-quinoxalinylcarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhex Acid, 2-L- (N-benzyloxycarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-7-methyl-1-phenyloctane, 2-L- (N-2-kinoline Carbonyl-L-asparaginyl) amino-epoxy-1-phenyloctane and the like.

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

Method I

In the above formulas, R ¹ , R ² , A and B are as defined above; P is a protecting group, Preferably it is a t-butoxycarbonyl group; Me is a methyl group; DIBAL is diisobutylaluminum hydride; Ph is a phenyl group; mCPBA is metachloroperoxybenzoyl acid.

According to Method I, first, a compound of formula (II) is esterified according to a conventional method to obtain a compound of formula (II), and then the compound of formula (III) is reduced to diisobutylaluminum hydride To obtain an aldehyde compound of formula (IV). Subsequently, the compound of general formula (IV) is reacted with the triphosphonium salt of general formula (V) by Wittig, and the compound of general formula (IV) is reacted with triphosphonium salt of general formula (V) by (Wittig) The compound of formula (VI) is obtained by reaction, and the reaction is preferably carried out in the presence of a base, and such base is preferably n-butyllithium, potassium carbonate, dimethyl sulfoxide anion or the like. After removing the protecting group from the compound of formula (VI) thus obtained, the compound of formula (VIII) is obtained by carrying out an amide coupling reaction with the compound of formula (VII). If it can be used conventionally, it can be used without a restriction | limiting in particular, Especially dicyclohexyl carbodiimide (DCC), 3-ethyl-3'- (dimethylamino) propyl carbodiimide (EDC), etc. can be used preferably. . The compound of formula (VIII) can then be epoxidized in metachloroperoxybenzoyl acid to afford the desired compound of formula (I).

Method II

In the above formulas, R ¹ , R ² , A and B are as defined above; P is a protecting group, preferably benzyloxycarbonyl; mCPBA is metachloroperoxybenzoyl acid.

In method II, the compound of formula (VI) obtained by a method similar to that of method I above is first epoxidized to give a compound of formula (IX), and then the protecting group is removed from the compound of formula (IX) And amide coupling reaction with a compound of the general formula (VII) to obtain a compound of the general formula (I). At this time, the epoxidation reaction and the amide coupling reaction are carried out in the same manner as in Method I.

In the method II, the amide coupling reaction can be performed twice as follows when m and n are both 1.

Wherein R ¹ , R ² , R ⁶ , R ⁷ , A and B are as defined above; P and P 'are each a protecting group, Preferably they are a benzyloxycarbonyl group.

That is, after removing a protecting group from the compound of general formula (IX), it carries out amide coupling reaction with the compound of general formula (X), obtaining the compound of general formula (XI), and then removing a protecting group from this compound of general formula (IX). Then, an amide coupling reaction with a carboxylic acid of formula (XII) or an acyl chloride of formula (XII ′) yields a compound of formula (I) wherein m and n are both 1. At this time, the reaction of coupling with the compound of the general formula (X) and the compound of the general formula (XII) can be carried out in the same manner as described above.

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

Compounds of the present invention have a total daily dose of 0.001-10 mg / kg body weight to be administered to the host, which may be administered at one time or several times, and specific dose levels for a particular patient may be determined by the specific compound to be used, weight, age, sex, health It may vary depending on the condition, diet, time of administration, method of administration, rate of excretion, drug mixture and severity of the 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 may be used include water, Ringer's solution, and isotonic NaCl solution, and sterile fixed oils are commonly compatible as solvents or suspending media.

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 slate 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), didioxyinosine (DDI), ribavirin, compound Q, alpha interferon, beta interferon, polymanno acetate, sodium phosphonophore Mate, HAD-23, UA001, eflornithine, acyclovir.

Area modifiers that can be administered in admixture with the compounds of the invention include bropyrimin, amprigen, anti-human alpha interferon antibody cloni stimulatory factor, CL246738, impe 2-2, diethyldithiocarbamate, interleukin-2 , Alpha-interferon, inosinepranovex, methionine enkephalin, muramyl-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 may be included as long as they are useful agents of the compounds of the present invention necessary for the treatment and prevention of HIV infection.

Such a present invention will be described in more detail based on examples. The following examples are only intended to aid 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: N-t-butoxycarbonyl-L-phenylalanine methyl ester

8.8 g (0.0344 mol) of N-t-butoxycarbonylphenylalanine was dissolved in 50 ml of dichloromethane, 50 mg of triethylamine was added at 0 ° C., and 3 ml of chloromethylformate was slowly added thereto. 50 mg of 4-dimethylaminopyridine was added to promote the reaction. After completion of the reaction, the reaction mixture was extracted with dichloromethane and distilled under reduced pressure to obtain the target compound quantitatively.

¹ H NMR (CDCl ₃ ) δ 1.4 (9H, s), 3.1 (2H, m), 3.7 (3H, s), 4.6 (1H, m), 5.0 (1H, d), 7.1 ~ 7.3 (5H, m )

Step 1: N-t-butoxycarbonyl-L-phenylalanal

1.3 g of the compound obtained in step 1 was dissolved in 20 ml of anhydrous toluene and the temperature was lowered at -78 ° C using dry ice-acetone. Diisobutylaluminum hydride was dissolved in hexane at 1 molar concentration under nitrogen pressure, and slowly added for 2 hours for 10 ml of the solution. After further stirring for about 50 minutes, excess diisobutylaluminum hydride was removed with 5 ml of methanol at -78 ° C, raised to room temperature, and extracted with ethyl acetate. After drying over anhydrous MgSO ₄ and concentrated to give the title compound in 95% yield. Used for the next reaction without further purification.

¹ H NMR (CDCl ₃ ) δ 1.4 (9H, s), 3.1 (2H, m), 4.4 (1H, m), 5.2 (1H, d), 5.0 (1H, d), 7.1 ~ 7.3 (5H, m ), 9.8 (1H, s)

Step 3: 3- (phenylpropyl) triphenylphosphine bromide

19.9 g (0.1 mol) of 1-bromo-3-phenylpropane and 26.2 g (0.1 mol) of triphenylphosphine were dissolved in 300 ml of anhydrous toluene and heated at reflux overnight. After cooling the reaction solution, the resulting solid was filtered, washed with 100 ml of diethyl ether and dried to give 28.1 g (60%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.92 (2H, m), 3.03 (2H, m), 3.85 (2H, br), 7.12 ~ 7.29 (5H, m), 7.60 ~ 7.84 (15H, m)

Step 4: 2-L- (t-butoxycarbonyl) amino-1.6-diphenylhex-3-cis-ene

18.5 mg (0.04 mmol) of the phosphine salt obtained in step 3 was dissolved in 2 ml of 1,4-dioxane, and 12.2 ml (0.08 mmol) of powdered K ₂ CO ₃ was added and stirred for 30 minutes. 10.0 mg (0.04 mmol) of the compound obtained in Example 2 was added thereto, followed by stirring at 90 ° C to 100 ° C for 30 minutes. The reaction solution was cooled, filtered to remove solids, diluted with 20 ml of diethyl ether, and washed twice with 10 ml of distilled water. After drying over anhydrous Mg _S O ₄ and distilling under reduced pressure, the residue was purified by column chromatography (hexane: ethyl acetate = 6: 1) to obtain 9.3 mg (66%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.39 (9H, br), 2.15 ~ 2.08 (6H, m), 4.30 ~ 4.62 (2H, br), 5.17 (1H, t), 5.42 (1H, m), 7.05 ~ 7.31 (10H, m)

Mass (FAB, m / e) 352 (M + 1)

Step 5: 2-L- (t-butoxycarbonyl) amino-3,4-epoxy-1,6-diphenylhexane

9.1 mg (0.026 mmol) of the compound obtained in step 3 was dissolved in 3 ml of dichloroethane, and 26.8 mg (content 50%, 0.078 mmol) of metachloroperoxybenzoic acid were added thereto and refluxed. It was refluxed again for 1 hour, cooled to room temperature and filtered to remove solids. After distillation under reduced pressure, the resulting oil was dissolved in 20 ml of ethyl acetate, washed twice with 10 ml of saturated Na ₂ S ₂ O ₃ solution, twice with 10 ml of saturated NaHCO ₃ solution, and twice with 10 ml of distilled water. After drying over anhydrous MgSO ₄ and distillation under reduced pressure, the residue was purified by column chromatography (hexane: ethyl acetate = 4: 1) to obtain 5.7 mg (60%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.39 (11H, br), 2.40 ~ 3.05 (6H, m), 3.70 (1H, br), 4.80 (1H, br), 7.02 ~ 7.34 (10H, m)

Example 2

Step 1: 2-L-amino-1,6-diphenylhex-3-ene trifluoroacetate salt

120 mg of the compound obtained in step 4 of Example 1 was dissolved in 3 ml of dichloromethane, 2 ml of trifluoroacetic acid was added thereto, stirred at room temperature for 30 minutes, and then distilled under reduced pressure to obtain the title compound quantitatively.

¹ H NMR (CDCl ₃ ) δ 2.7 (2H, m), 3.0 (2H, t), 3.2 ~ 3.5 (2H, m), 4.6 (1H, br), 5.9 (1H, m), 6.3 (1H, m ), 7.4-8.0 (10H, m), 10.5 (3H, br)

Step 2: 2-L- (N-benzyloxycarbonyl-L-asparaginyl) amino-1,6-diphenylhex-3-cis-ene

17 mg (0.067 mmol) of N-benzyloxycarbonyl-L-asparagine, 15 mg of 3-ethyl-3 '-(dimethylamino) propylcarbodiimide, and 10 mg of N-hydroxybenzotriazole were dissolved in anhydrous dimethylformamide 30 After stirring for 10 minutes, 18 mg (0.05 mmol) of the compound obtained in Step 1 and 10 ul of N-methylmorpholine were added thereto, followed by stirring at room temperature for 24 hours. Distilled under reduced pressure, dimethylformamide was dissolved in a mixture of 30ml of ethyl acetate, and then washed three times with 1N HCl and saturated aqueous NaHCO ₃ solution. After drying over anhydrous MgSO ₄ and removing the solvent, the residue was purified by column chromatography (ethyl acetate: hexane = 2: 5) to give 14.5 mg (60%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, m), 2.0 (1H, m), 2.2 ~ 2.9 (6H, m), 3.9 (1H, m), 4.9 (1H, m), 5.2 (1H, m ), 5.3 (1H, d), 5.5 (1H, m), 5.8 (1H, d), 7.1-7.4 (15H, m)

Step 3: 2-L- (N-benzyloxycarbonyl-L-asparaginyl) amino-3R, 4S) -epoxy-1,6-diphenylhexane

The compound obtained in step 2 was reacted in the same manner as in step 5 of Example 1, and then purified by column chromatography (ethyl acetate: hexane = 1: 2) to obtain the title compound in a yield of 30%.

¹ H NMR (CDCl ₃ ) δ 1.5 (2H, m), 2.4 ~ 3.0 (8H, m), 3.9 (1H, m), 4.4 (1H, m), 5.1 (2H, s), 5.4 (1H, d ), 5.6 (1H, d), 6.4 (2H, br), 7.0-7.4 (15H, m)

Mass (FAB, m / e) 516 (M + 1)

Example 3

Step 1: 2-L- (N-benzyloxycarbonyl-L-valanyl) amino-1,6-diphenylhex-3-cis-ene

The title compound was synthesized in 70% yield by the same procedure as in step 2 of Example 2 above using N-benzyloxycarbonyl-L-valine instead of N-benzyloxycarbonyl-L-asparagine.

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, m), 2.0 (1H, m) 1, 2.2 ~ 2.9 (6H, m), 3.9 (1H, m), 4.9 (1H, m), 5.1 (2H, m), 5.2 (1H, s), 5.3 (1H, d), 5.5 (1H, m), 5.8 (1H, d), 7.1-7.4 (15H, m)

Step 2: 2-L- (N-benzyloxy-L-balanyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The compound obtained in Step 1 was reacted in the same manner as in Step 3 of Example 2, to obtain the title compound.

¹ H NMR (CDCl ₃ ) δ 0.9 (6H, m), 1.4 (1H, m), 1.6 (1H, m), 2.1 (1H, m), 2.5 ~ 3.0 (6H, m), 4.0 (2H, m ), 5.1 (2H, s), 5.3 (1H, d), 6.1 (1H, d), 7.1 ~ 7.4 (15H, m)

Mass (FAB, m / e) 501 (M + 1)

Example 4

Step 1: 2- (N-benzyloxycarbonyl-isorocinyl) amino-1,6-diphenylhex-3-cis-ene

The title compound was synthesized in 70% yield by the same procedure as in step 2 of Example 2 above using N-benzyloxycarbonyl-L-isorocin instead of N-benzyloxycarbonyl-L-asparagine.

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, m), 1.0 ~ 1.5 (2H, m), 1.8 (1H, m), 2.2 ~ 2.8 (6H, m), 3.9 (1H, m), 4.9 (1H , s), 5.1 (2H, s), 5.2 (2H, m), 5.5 (1H, m), 5.8 (1H, d), 7.1 ~ 7.4 (15H, m)

Step 2: 2-L- (N-benzyloxycarbonyl-L-isorocinyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The compound obtained in Step 1 was reacted in the same manner as in Step 3 of Example 2, to obtain the title compound.

¹ H NMR (CDCl ₃ ) δ 0.9 (6H, m), 1.0 ~ 1.7 (4H, m), 1.9 (1H, m), 2.5 ~ 3.1 (6H, m), 4.0 (2H, m), 5.1 (2H s), 5.3 (1H, d), 6.1 (1H, d), 7.1 ~ 7.4 (15H, m)

Mass (FAB, m / e) 513 (M + 1)

Example 5

Step 1: 2- (N-benzyloxycarbonyl-L-glutaminyl) amino-1,6-diphenylhex-3-cis-ene

The title compound was obtained in 60% yield by the same method as in step 2 of Example 2 above using N-benzyloxycarbonyl-L-glutamine instead of N-benzyloxycarbonyl-L-asparagine.

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, m), 2.0 (1H, m), 2.2 ~ 2.9 (8H, m), 3.9 (1H, m), 5.0 (1H, m), 5.1 (2H, m ), 5.2 (1H, m), 5.3 (1H, d), 5.5 (1H, m), 5.8 (1H, d), 7.1 ~ 7.4 (15H, m)

Step 2: 2-L- (N-benzyloxycarbonyl-L-glutaminyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The compound obtained in Step 1 was reacted in the same manner as in Step 3 of Example 2, to obtain the title compound in a yield of 35%.

¹ H NMR (CDCl ₃ ) δ 1.5 (2H, m), 2.4 ~ 3.0 (10H, m), 3.9 (1H, m), 4.3 (1H, m), 5. 2 (2H, s), 5.4 (1H d), 5.6 (1H, d), 6.4 (2H, br), 7.0-7.4 (15H, m)

Mass (FAB, m / e) 530 (M + 1)

Example 6

2-L- (N-pepoxyacetyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

25 mg of 2-L- (N-pexoxyacetyl-L-asparaginyl) amino-1,6-diphenyl-hex-3-x-ene was dissolved in 5 ml of dichloromethane and The reaction was carried out in the same manner, and then purified by column chromatography (ethyl acetate: hexane = 1: 1) to obtain 3 mg (20%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.2 ~ 1.6 (2H, m), 2.4 ~ 3.2 (8H, m), 4.4 (2H, s), 4.7 (1H, m), 6.4 (2H, br), 7.0 ~ 7.5 (15H, m)

Mass (FAB, m / e) 516 (M + 1)

Example 7

2-L- [N- (2,4-difluoro) phenoxyacetyl-L-asparaginyl] amino- (3R, 4S) -epoxy-1,6-diphenylhexane

25 mg of 2-L- [N- (2,4-difluoro) phenoxyacetyl-L-asparaginyl] amino-1,6-diphenyl-hex-3-cis-ene was dissolved in 5 ml of dichloromethane. After reacting in the same manner as in Step 3 of Example 2, the residue was purified by column chromatography (ethyl acetate: hexane = 1: 1) to obtain 5 mg (yield 30%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.2 ~ 1.6 (2H, m), 2.4 ~ 3.2 (8H, m), 3.9 (1H, m), 4.4 (2H, s), 4.7 (1H, m), 6.4 (2H , br), 7.0-7.5 (13H, m)

Mass (FAB, m / e) 522 (M + 1)

Example 8

Step 1: N-benzyloxycarbonyl-L-phenylalanine methyl ester

The title compound was obtained in the same manner as in Example 1 using N-benzyloxycarbonyl phenylalanine.

Step 2: N-benzyloxycarbonyl-L-phenylalanal

6.0 g (0.019 mol) of the compound obtained in step 2 was added to anhydrous toluene solution and cooled to -50 ° C using dry ice-acetone. While stirring vigorously under nitrogen pressure, 22 ml of a solution obtained by dissolving diisobutylaluminum hydride in 1.76 mol concentration in hexane was taken, and slowly added over 30 minutes. After a further 20 minutes of stirring, excess diisobutylaluminum hydride was removed by addition of 2N HClDYDDOR. The temperature of the reaction solution was raised to 0 ° C. and extracted twice with 60 ml of ethyl acetate. The combined organic layers were washed with saturated NaCl solution, dried over anhydrous MgSO ₄ , and distilled under reduced pressure to obtain 3.8 g of the target compound. Used for the next reaction without further purification.

¹ H NMR (CDCl ₃ ) δ 3.05-3.20 (2H, d), 4.52 (1H, q), 5.10 (2H, s), 5.31 (1H, br), 7.10-7.43 (10H, m), 9.63 (1H , s)

Mass (FAB, m / e) 284 (M + 1)

Step 3: 2-L- (N-benzyloxycarbonyl) amino-1,6-diphenylhex-3-ene

The compound obtained in Step 2 was reacted in the same manner as in Step 4 of Example 1, to obtain the title compound (cis: trans = 9: 1) in a yield of 60%. Purified by column chromatography (hexane: ethyl acetate = 6: 1).

Step 4: 2-L- (benzyloxycarbonyl) amino-3,4-epoxy-1,6-diphenylhexane

3.3 g (8.56 mole) of the compound obtained in step 3 was dissolved in 50 ml of chloroform, and then 2.94 g (content 50%, 17.1 mmol) of metachloroperoxybenzoic acid were added and refluxed. After refluxing for 3 hours, the mixture was cooled to room temperature, and washed twice with 20 ml of 10% Na2S2O2 solution, 20 ml of saturated NaHCO3 solution, and 20 ml of distilled water. Dry over anhydrous MgSO4 and purify by column chromatography (hexane: ethyl acetate = 6: 1) to give a total of 2.57 g (75%) of the product 2- (benzyloxycarbonylamino)-(3R, 4S) -epoxy-1 , 6-diphenylhexane: 2- (benzyloxycarbonylamino)-(3R, 4S) -epoxy-1,6-diphenylhexane: 2- (benzyloxycarbonylamino)-(3R, 4S) -epoxy -1,6-diphenylhexane was obtained by the ratio of 7: 2: 1.

A) 2-L- (benzyloxycarbonyl) amino- (3S, 4R) -epoxy-1,6-diphenylhexane

¹ H NMR (CDCl ₃ ) δ 1.32-1.70 (2H, m), 2.46-3.08 (6H, m), 3.7 (1H, m), 5.0 (1H, d), 5.1 (2H, s), 7.05-7.4 (15H, m)

Mass (FAB, m / e) 402 (M + 1)

B) 2-L- (benzyloxycarbonyl) amino- (3S, 4R) -epoxy-1,6-diphenylhexane

¹ H NMR (CDCl ₃ ) δ 1.78 (2H, m), 2.58 ~ 3.0 (6H, m), 4.1 (1H, m), 4.75 (1H, d), 5.05 (2H, s), 7.1 ~ 7.4 (15H , m)

Mass (FAB, m / e) 402 (M + 1)

Example 9

Step 1: 2-amino- (3S, 4R) -epoxy-1,6-diphenylhexane

0.4 (g) (1 mmol) of Compound (A) obtained in Step 4 of Example 8 was dissolved in 10 ml of anhydrous methanol, and 40% of 10% Pb / C was mixed and stirred for 6 hours at room temperature under hydrogen gas pressure. After filtration through celite, methanol was distilled off under reduced pressure to obtain 0.26 g (95%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.3 ~ 1.6 (2H, m), 2.35 ~ 3.0 (7H, m)

Step 2: 2-L- (N-benzyloxycarbonyl-L-asparaginyl) amino- (3S, 4R) -epoxy-1,6-diphenylhexane

0.26 g (1 mmol) of the compound obtained in Step 1, 0.27 g (1 mmol) of N-benzyloxycarbonyl-L-asparagine, and 0.29 g (1.5 mmol) of 3-ethyl-3 '-(dimethyl amino) propylcarbonylimide Mmol), 0.21 g (1.5 mmol) of N-hydrobenzotriazole and 0.15 g (1.5 mmol) of triethylamine were dissolved in 8 ml of anhydrous dimethylformamide, dissolved at 40 ° C, and reacted with stirring for 15 hours. After completion of the reaction, dimethylformamide was removed by distillation under reduced pressure, dissolved in 50 ml of ethyl acetate, washed with water and treated with anhydrous MgSO ₄ . The solvent was removed and separated by column chromatography (ethyl acetate 100%) to yield 0.26 g (50%) of the title compound. NMR and mass spectrum were the same as the result obtained in step 3 of Example 2 above.

Example 10

2-L- [N- (2,4-difluoro) phenoxyacetyl-L-asparaginyl] amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The benzyloxycarbonyl protecting group was removed from the compound obtained in step 2 of Example 9 in the same manner as in step 1 of Example 9, and the compound was then replaced by N-benzyl instead of N-benzyloxycarbonyl-L-asparagine. Reaction was carried out in the same manner as in Step 2 of Example 9, except that oxycarbonyl-L-alanyl was used to obtain the title compound in yield.

¹ H NMR (CD ₃ OD) δ 1.2 ~ 1.5 (2H, m), 1.35 (3H, d), 2.4 ~ 3.1 (8H, m), 3.9 (1H, m), 4.1 (1H, m), 4.7 ( 1H, m), 5.12 (2H, s), 7.05 ~ 7.4 (15H, m)

Mass (FAB, m / e) 587 (M + 1)

Example 11

2- (N-2-Quinolinecarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

100 mg (0.2 mmol) of the compound obtained in step 2 of Example 9 was dissolved in 2 ml of anhydrous methanol, and 10 mg of 10% Pd / C was mixed and stirred for 6 hours at room temperature under hydrogen gas pressure. After filtration through celite, the methanol was removed, 41 mg (0.24 mmol) of 2-quinolinecarboxylic acid, 38 g (0.2 mmol) of 3-ethyl-3 '(dimethylamino) propylcarbodiimide, N-hydroxybenzotriazole It was dissolved in 27 mg (0.2 mmol), 24 mg (0.24 mmol) of triethylamine and 3 ml of anhydrous dimethylformamide and stirred at about 40 ° C. for 15 hours. After completion of the reaction, dimethylformamide was removed and then dissolved in ethyl acetate. After washing several times with water and dried over anhydrous MaSO 4 and distilled under reduced pressure, the residue was separated by column chromatography (ethyl acetate: hexane = 1: 1) to obtain 40 mg (yield 37%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.4-1.7 (2H, m), 2.3-3.0 (8H, m), 4.1 (1H, m), 5.0 (1H, m), 5.5 (1H, br), 6.0 (1H , br), 6.7 (1H, s), 7.05-8.3 (16H, m), 9.5 (1H, dd)

Mass (FAB, m / e) 537 (M + 1)

Example 12

2- (N-1-naphthoxyacetyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The title compound was obtained in 65% yield by the same method as in Example 11 above using 1-naphthoxyacetyl acid instead of 2-quinolinecarboxylic acid.

¹ H NMR (CDCl ₃ ) δ 1.4 ~ 1.7 (2H, m), 2.3 ~ 3.0 (8H, m), 4.0 (1H, m), 4.6 (2H, s), 4.9 (1H, m), 5.6 (1H d), 6.1 (1H, s), 6.8 ~ 7.8 (1H, m), 8.4 (1H, t), 8.6 ~ 8.7 (1H, dd)

Mass (FAB, m / e) 566 (M + 1)

Example 13

2- (N-1-quinoxalin-2-ylcarbonyl-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

150 mg (0.29 molmol) of the compound obtained in step 2 of Example 9 was removed in the same manner as in step 1 of Example 9, followed by 73 mg (0.38 of quinoxalin-2-yl carbonyl chloride). Mmol) and triethylamine 146 mg (1.45 mmol) were dissolved in anhydrous dichloromethane and stirred at reflux for 10 hours. After completion of the reaction, the mixture was washed twice with saturated NaHCO 3 aqueous solution, dried over anhydrous MgSO 4, and distilled under reduced pressure. Column chromatography (ethyl acetate 100%) was isolated to give 54 mg (yield 34%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.3 ~ 1.8 (2H, m), 2.4 ~ 3.1 (8H, m), 4.05 (1H, m), 4.95 (1H, m), 5.5 (1H, br), 5.9 (1H , d), 7.1 to 7.4 (10H, m), 7.5 (1H, d), 7.9 (1H, m), 8.2 (1H, m), 9.2 (1H, dd), 9.6 (1H, s)

Mass (FAB, m / e) 538 (M + 1)

Example 14

Step 1: N-morphulincarbonyl-L-alanine

2 g (50 mmol) of NaOH and 30 ml of distilled water were added to 4 g (50 mmol) of L-alanine, and cooled to 0 ° C., followed by vigorous stirring. 7.1 g (50 mmol) of morpholin carbonyl chloride was dissolved in 30 ml of tetrahydrofuhan and slowly added to pH 8.5-pH 9.5 with 0.05 N NaOH. After completion of the reaction, the mixture was lowered to pH 2 with 6N HCl, extracted three times with 30 ml of a mixed solvent of ethyl acetate and sol (1: 1), and distilled under reduced pressure to obtain 4 g (40%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 1.3 (3H, d), 3.2 (4H, br), 3.6 (4H, br), 4.0 (1H, q)

Step 2:

The compound obtained in Step 1 of Example 10 and the compound obtained in Step 1 were coupled in the same manner as in Step 2 of Example 9, and then purified by column chromatography (ethyl acetate = 100%) to obtain the title compound. Yield 58.4%.

¹ H NMR (CDCl ₃ ) δ 1.2 ~ 1.7 (5H, m, d), 2.4 ~ 3.1 (8H, m), 3.4 (4H, br), 3.7 (4H, br), 3.9 (1H, m), 4.2 (1H, m), 4.7 (1H, m), 5.3 (1H, br), 5.7 (1H, d), 6.2 (1H, br), 7.0 ~ 7.5 (10H, m), 7.8 (1H, dd), 8.0 (1H, d)

Mass (FAB, m / e) 565 (M + 1)

Example 15

2-L- [N- (N-phenylalaninyl) -L-asparaginyl] amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The title compound was synthesized in the yield of 54% according to Examples 1 and 2, except that L-phenylalanine was used instead of L-alanine in Step 1 of Example 12.

¹ H NMR (CDCl ₃ ) δ 1.2 ~ 1.7 (5H, m), 2.4 ~ 3.2 (10H, m), 3.4 (4H, br), 3.7 (4H, br), 5.7 (1H, d), 6.2 (1H , br), 7.0 to 7.5 (15H, m), 7.6 (1H, dd), 7.9 (1H, d)

Example 16

2-L- (N-3-triazolopancarbonyl-L-asparaginyl] amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The benzyloxycarbonyl group was removed from the compound obtained in step 2 of Example 9 in the same manner as in step 1 of Example 9, and the compound was then substituted with 3-triazole instead of N-benzyloxycarbonyl-L-asparagine. The title compound was obtained in 58% yield by the same method as in step 2 of Example 9, except for using propyl acid.

¹ H NMR (CD ₃ OD) δ 1.2 ~ 1.6 (2H, m), 2.4 ~ 3.2 (10H, m), 3.9 (1H, m), 4.5 (2H, t), 4.75 (1H, m), 7.0 ~ 7.4 (10H, m), 8.0 (1H, s), 8.45 (1H, s)

Mass (FAB, m / e) 505 (M + 1)

Example 17

2-L- [N-3-triazolopancarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane

The benzyloxycarbonyl group was removed from the compound obtained in step 2 of Example 9 in the same manner as in step 1 of Example 9, and the compound was then replaced with N-benzyloxy instead of N-benzyloxycarbonyl-L-asparagine. The title compound was obtained in 65% yield by the same method as in step 2 of Example 7, except that carbonyl-L-phenylalanyl was used.

¹ H NMR (CD ₃ OD) δ 1.2 to 1.5 (2H, m), 2.4 to 3.1 (10H, m), 3.9 (1H, m), 4.3 (2H, m), 4.7 (1H, m), 5.05 ( 2H, s), 7.0 ~ 7.5 (20H, m)

Example 18

2-L- [N- (N-morphopropionyl-L-phenylalanyl) L-asparaginylamino- (3R, 4S) -epoxy-1,6-diphenylhexane

The benzyloxycarbonyl group was removed from the compound obtained in Example 17 in the same manner as in Step 1 of Example 9, and then 3-morpholine propionic acid was used instead of N-benzyloxycarbonyl-L-asparagine. The title compound was obtained after the reaction in the same manner as in Step 2 of Example 9, except that.

Mass (FAB, m / e) 670 (M + 1)

Example 19

Step 1: 2-L- (N-2-quinolinecarbonyl-L-asparaginyl) amino-1,6-diphenyl-hex-3-cis-ene

Example 9 The compound obtained in Step 1 of Example 9 was replaced with N-benzyloxycarbonyl-L-asparagine except that N- (quinolin-2-yl-carbonyl) -L-asparagine was used. The reaction was carried out in the same manner as in step 2, followed by column chromatography (ethyl acetate: hexane = 1: 2) to obtain 60 mg (yield 40%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 2.2-3.1 (8H, m), 4.7 (2H, m), 5.2 (1H, t), 5.4 (1H, m), 6.4 (1H, d), 7.0-7.5 (10H , m), 7.6-8.3 (6H, m), 8.8 (1H, dd)

Step 2:

20 mg of the compound obtained in step 1 was dissolved in 5 ml of dichloromethane and reacted in the same manner as in step 4 of Example 8, followed by column chromatography (ethyl acetate: hexane = 1: 1) to obtain 45 mg of the title compound (yield). 25%).

¹ H NMR (CDCl ₃ ) δ 1.2 ~ 1.6 (2H, m), 2.4 ~ 3.2 (8H, m), 3.9 (1H, m), 5.0 (1H, m), 6.8 (1H, d), 7.0 ~ 8.3 (16H, m), 8.8 (1H, dd)

Mass (FAB, m / e) 553 (M + 1)

Example 20

Step 1: N-t-butoxycarbonyl-L-leucine methyl ester

The title compound was obtained in the same manner as in Step 1 of Example 1, using N-n-butoxycarbonyl leucine instead of N-t-butoxycarbonylphenylalanine.

Step 2: N-t-butoxycarbonyl-L-leucine

The compound obtained in Step 1 was reacted in the same manner as in Step 2 of Example 1, to obtain the title compound.

Step 3: 5-L- [N- (t-butoxycarbonyl)] amino-7-methyl-1-phenyl-oct-3-cis-ene

215 mg (1 mmol) of the compound obtained in Step 2 and 262 mg (1 mmol) of the compound obtained in Step 3 of Example 1 were added to 200 mg of K ₂ CO ₃ and 10 ml of 1,4-dioxane and refluxed for 4 hours. After completion of the reaction, the precipitate was filtered off, and 1,4-dioxane was distilled under reduced pressure, followed by extraction with 100 ml of diethyl ester. After drying over MgSO ₄ and distillation under reduced pressure, the residue was purified by column chromatography (ethyl acetate: hexane = 1: 8) to obtain 300 mg (95%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.9 (6H, 2d), 1.1 ~ 1.7 (3H, m), 1.4 (9H, m), 2.3 ~ 2.8 (4H, m), 4.2 ~ 4.5 (2H, m), 5.2 (1H, m), 5.5 (1H, m), 7.1-7.5 (5H, m)

Mass (FAB, m / e) 318 (M + 1)

Step 4: 5-L-amino-7-methyl-1-oct-3-cis-ene trifluoro acetate

42 mg (0.13 mmol) of the compound obtained in step 3 was dissolved in 1 ml of dichloromethane, trifluoroacetic acid was added thereto, stirred at room temperature for 30 minutes, and distilled under reduced pressure to obtain the title compound quantitatively.

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, 2d), 1.3 ~ 1.5 (3H, m), 2.4 (2H, m), 2.7 (2H, m), 3.9 (1H, br), 5.3 (1H, m ), 5.8 (1H, m), 7.0 ~ 7.3 (5H, m), 10.5 (3H, br)

Step 5: 5-L- (N-benzyloxycarbonyl-L-asparag) amino-7-methyl-1-phenyloct-3-cis-ene

The compound obtained in step 4 was reacted in the same manner as in step 2 of Example 9, and then purified by column chromatography (ethyl acetate: hexane = 1: 2) to obtain 24 mg (40%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.9 (6H, 2d), 1.1 ~ 1.56 (3H, m), 2.3 ~ 3.0 (6H, m), 4.4 (1H, m), 4.7 (1H, m), 5.1 (2H , s), 5.2 (1H, m), 5.5 (2H, m), 6.0 (1H, d), 7.1 ~ 7.4 (10H, m)

Mass (FAB, m / e) 466 (M + 1)

Step 6: 5-L- (N-benzyloxycarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-7-methyl-1-phenyloctane

10 mg of the compound obtained in step 5 was dissolved in dichloromethane, and then reacted in the same manner as in step 4 of Example 8, followed by purification by column chromatography (ethyl acetate: hexane = 1: 2) to 5 mg of the title compound. (Yield 50%) was obtained.

¹ H NMR (CDCl ₃ ) δ 0.9 (6H, dd), 1.1 ~ 1.6 (3H, m), 1.8 (2H, m), 2.7-3.1 (6H, m), 3.9 (1H, m), 4.5 (1H , m), 5.1 (2H, s), 5.6 (1H, d), 6.3 (1H, d), 7.1-7.4 (10H, m)

Mass (FAB, m / e) 482 (M + 1)

[Example 21]

Step 1: 2-L- (benzyloxycarbonylamino) 7-methyl-1-phenyl-oct-3-cis-ene

2.49 g (10 mmol) of L-phenylalanal and 4.27 g (10 mmol) of 4-methyl-1-pentyltriphenylphosphonum bromide are dissolved in 50 ml of 1,4-dioxane, and 2.76 g of powdered K ₂ CO ₃ ( 20 mmol) was added and stirred at the reflux temperature of the reaction solvent for 5 hours. The reaction solvent was cooled and filtered to remove solids. 30 ml of diethyl ether was added, diluted, and washed with 10 ml of distilled water. After drying over anhydrous MaSO ₄ and distillation under reduced pressure, the residue was purified by column chromatography (hexane: ethyl acetate = 10: 1) to obtain 2.02 g (yield 60%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, d), 1.1 ~ 1.5 (5H, m), 1.8 (1H, m), 2.8 (2H, m), 5.0 (2H, s), 5.05 ~ 5.3 (2H , m), 7.0 to 7.4 (10H, m)

Step 2: 2-L- (benzyloxycarbonylamino)-(3R, 4S) -epoxy-7-methyl-1-phenyl-octane

2.02 g of the compound obtained in step 1 was dissolved in 20 ml of chloroform and reacted in the same manner as in step 4 of Example 8, followed by purification by column chromatography (ethyl acetate: hexane = 1: 2) in total, 0.8 g (yield). 50%) of product was obtained, of which the title compound was 0.6 g (40% yield).

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, dd), 1.0 ~ 1.5 (5H, m), 2.8 ~ 3.1 (4H, m), 3.8 (1H, m), 5.05 (2H, s), 7.1 ~ 7.4 (10H, m)

Example 22

2-L- (N-benzyloxycarbonyl-L-asparagyl) amino- (3R, 4S) -epoxy-7-methyl-1-phenyl-octane

500 mg (1.36 mmol) of the compound obtained in Example 21 were removed in the same manner as in Step 1 of Example 9, and then the compound was reacted in the same manner as in Step 2 of Example 9 to obtain the title compound. 400 mg (yield 55%).

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, dd), 0.9 ~ 1.4 (5H, m), 2.4 ~ 3.0 (6H, m), 3.8 (1H, m), 5.0 (2H, s), 7.0 ~ 7.4 (10H, m)

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

Example 23

2-L- [N- (2-quinolinecarbonyl) -L-asparagyl) amino- (3R, 4S) -epoxy-7-methyl-1-phenyl-octane

150 mg (0.31 mmol) of the compound obtained in Example 21 were coupled in the same manner as in Example 11, and then separated by column chromatography (100% ethyl acetate), to obtain 60 mg (yield 40%) of the title compound.

¹ H NMR (CDCl ₃ ) δ 0.8 (6H, dd), 0.9 ~ 1.6 (5H, m), 2.6 ~ 3.0 (6H, m), 4.0 (1H, m), 4.9 (1H, m), 5.4 (1H , br), 6.0 (1H, br), 7.0 ~ 7.3 (5H, m), 7.4 (1H, br), 7.6 ~ 8.3 (6H, m), 9.3 (1H, dd)

Mass (FAB, m / e) 503 (M + 1)

Example 24

Step 1: N-morpholinesulfonyl chloride

16 g of concentrated hydrochloric acid, 16 g of water, and 14.2 g of morpholine were mixed, the temperature was lowered to 5 ° C., and 80 ml of dichloromethane was added. 315 g of 4% NaOCl was added at 5 ° C. over 60 minutes, and then stirred at room temperature for 50 minutes. The dichloromethane layer was separated, dried over MgSO ₄ , the dichloromethane solution was cooled to −70 ° C., and then 24 g of SO ₂ and 100 ml of dichloromethane were further added. 3 ml of chlorine gas was condensed into the mixture, and the mixture was stirred while gradually raising the temperature to room temperature for 24 hours. The mixture was washed with pH 7.0 phosphate buffer solution (0.25 molarity), washed until the pH of the washed water was neutral, and the dichloromethane layer was dried over MgSO ₄ . Distillation under reduced pressure afforded the title compound in a yield of 60%.

¹ H NMR (CDCl ₃ ) δ 1.2 ~ 1.7 (5H, m, d), 2.4 ~ 3.2 (12H, m), 3.7 (4H, br), 3.9 (2H, m), 4.7 (1H, m), 6.8 (1H, br), 6.9 (1H, br), 7.05 ~ 7.4 (10H, m), 7.5 (1H, d), 7.7 ~ 7.8 (1H, dd), 8.3 ~ 8.5 (1H, dd)

Mass (FAB, m / e) 602 (M + 1)

Step 2:

The benzyloxycarbonyl protecting group was removed from the compound obtained in Example 10 in the same manner as in Step 1 of Example 9, and then the compound was stirred with N-morpholinesulfonyl chloride at room temperature for 4 hours. After distillation under reduced pressure, column chromatography with 100% ethyl acetate gave the title compound in 70% yield.

Mass (FAB, m / e) 634 (M + 1)

Assays for Screening the Inhibitory Activity of HIV Proteases

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

An oligopeptide consisting of 11 amino acids of Ser-Nle-Ala-Glu- (p-NO ₂ ) -Pha-Seu-Val-Arg-Lys-His was used as the reactor. This substrate was subjected to HIV protease using a strong absorbance at 280 nm 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.

The compounds of the present invention are ultimately irreversible inhibitors having an inhibitory effect of 100%, and Table 1 shows the irreversible inhibition constants of certain compounds of the present invention.

TABLE 1

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. Here, the HIV-1 inoculum of 50 TCID ₅₀ was inserted and 3 days after SupT1, syncytia formation was examined. In the case of H9, three quarters of the culture medium was replaced with fresh culture medium containing the same concentration of inhibitor at three-day intervals. After 9 days, 1 ml was taken and centrifuged, and the supernatant was taken to measure the inhibition rate of HIV by antigen test (ELIZA) or reverse transcriptase search. IC ₅₀ is the concentration of compound at which point 50% of the cells can survive. Table 2 below shows the degree of inhibition in SupT1.

TABLE 2

Claims (4)

Cis-epoxide derivatives represented by the following general formula (I), pharmaceutically acceptable non-toxic salts thereof, isomers thereof, and hydrate solvates. Wherein R ¹ and R ² may be the same or different and each is a lower alkyl group or an arylalkyl group; A is R3- (CH ₂ ) ₁ Wherein R ³ is an aryl group, an aryloxy group substituted or unsubstituted with lower alkyl or halogen or hydroxy, an aromatic heterocyclic system containing 1 to 3 nitrogen atoms or an N-oxide system thereof, or oxygen and nitrogen A 3 to 6 membered heterocyclic system containing atoms at the same time or containing only nitrogen atoms, 1 being 0, 1 or 2), Wherein R 4 is a lower alkyl group or an arylalkyl group, or ; Wherein R ⁵ is a substituted heterocyclic ring system containing oxygen and nitrogen atoms; B is group represented by following General formula (a). (Wherein R ⁶ is a lower alkyl group or carbamoyl group, R ⁷ is a lower alkyl group or allylalkyl group, and m and n are each 0 or 1). A compound according to claim 1, wherein R ¹ is an isobutyl group or benzyl group; R ² is a 2-phenylethyl group or an isopentyl group; A is R _3- (CH ₂ ) ₁ R ³ is phenoxymethyl, 2,4-difluorophenoxymethyl, naphthoxymethyl, quinolinyl, oxyquinolinyl, morpholinyl, morpholinpropyl or quinoxalinyl, and 1 is 0, 1 or 2, R ⁴ is t-butyl or benzyl, or It is represented by and R <^5> is group which is morpholine; B is a compound in which asparagine, alanine, phenylalanine, glutamine or valine residues are alone or optionally two bonded residues. The 2-L- (t-butoxycarbonyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-benzyloxycarbo) according to claim 1 or 2 Nyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-benzyloxycarbonyl-L-glutaminyl) amino- (3R, 4S ) -Epoxy-1,6-diphenylhexane, 5-L- (N-benzyloxycarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-7-methyl-1-phenyloctane, 2 -L- (N-phenoxyacetyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-1-naphthoxyacetyl-L-aspa Laginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-2-quinolinecarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy- 1,6-diphenylhexane, 2-L- (N-quinoxalinecarbonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- [N -(N-morpholinesulfonyl-L-asparaginyl) amino- (3R, 4S) -epoxy-1,6-diphenylhexane, 2-L- (N-benzyloxycarbonyl-L-asparaginyl Amino- (3R, 4S) -epoxy-7-methyl-1-phenyl Tan, 2-L- (N-2- quinolin-carbonyl -L- asparaginyl) amino - (3R, 4S) - epoxy-1-phenyl octane and compound selected from the accepted non-toxic salts with pharmaceutically. A composition for the treatment or prevention of a human immunodeficiency virus infection containing a therapeutically effective amount of a compound according to claim 1 and containing at least one adjuvant selected from a pharmaceutically acceptable carrier, dispersant, wetting agent, inert diluent, excipient and lubricant. .