KR0138603B1 - Human hiv protease inhibitor, process for the preparation thereof and composition containing same - Google Patents

Abstract

The present invention provides a novel compound that inhibits human immunodeficiency virus (HIV) protease represented by the following general formula (I) and a method for preparing the same, and the treatment of acquired immunodeficiency syndrome caused by HIV infection containing the compound as an active ingredient. Or to a composition for prevention.

Where

A, D and R ₁ are as defined in the specification.

Description

Human Immunodeficiency Virus (HIV) Protease Inhibitory Compounds, Methods for Making the Same, and Compositions Comprising the Same

The present invention provides a novel compound that inhibits human immunodeficiency virus (HIV) protease, a method for preparing the same, and a composition for treating or preventing AIDS caused by HIV infection, comprising the compound as an active ingredient. It is about.

HIV-1 is a retrovirus that causes AIDS and AIDS syndrome, keeping genetic information in RNA form. The structure of the virus contains RNA, the innermost genetic information, and reverse transcriptase, which can generate DNA from the RNA, which is surrounded by the shell protein and on the outside. The lipid film forms a protective film. The reverse transcriptase is an enzyme that replicates DNA from RNA. Because of this specificity, the reverse transcriptase is an enzyme that exists only in RNA viruses, that is, retroviruses. Retroviruses infect the host and then make double-stranded DNA from the viral RNA by the reverse transcriptase, and the DNA thus prepared is integrated into the host's chromosome by integrase to create a new viral RNA that is infected as well as the host. By using the enzyme mechanism of the virus can be made. The resulting polyprotein is modified by enzymes of the host or enzymes of the virus to form new viruses. One of the most important enzymes causing the modification is protease, which is an enzyme that breaks down a polyprotein into structural proteins and enzymes necessary for virus formation.

Among the retrovirus proteases, HIV proteases have been the most intensive research subjects. As mentioned above, HIV does not synthesize each of them from mRNA when making bark proteins and necessary enzymes. Instead, Gag-protein (P55) is a long polyprotein that combines all of the enzymes into the bark protein. ) Or Gag-po1 protein (P165) is made first, and then the polyprotein is broken down into enzymes necessary for virus formation such as shell protein, reverse transcriptase, and integrase by proteases inside. Inhibiting the protease responsible for this degradation process stops the replication of the virus, which is based on this principle.

In conventional mutation experiments, viruses that do not have the function of such proteases have been reported to be not infectious [Kohl et al., Rroc. Nat. Acad. Sci., USA, 85, 4686-4690 (1988); and Peng et al., J. Virol., 63, 2550 (1989)]. Thus, inhibitors that inhibit the function of HIV proteases are produced by HIV infection. The possibility for use as a therapeutic agent for diseases has been suggested.

The HIV protease consists of 99 amino acids and its structure has been determined by X-ray structuring [Navia et al., Nature 337, 615-620 (1989); Wlodawer et al., Science, 245, 616-621 (1989); and Miller et al., Science, 246, 1149-1152). HIV protease is a typical aspartic protease in which two identical units are present as dimers and the molecular weight of each unit is 10793, with an arrangement 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 similar to the enzyme's transition state (TSA) to increase the affinity of the enzyme, thus enhancing its binding to the enzyme. HIV protease inhibitors developed in this manner have been described in several documents [Roberts, et al., Science, 248, 358 (1990); European Patent Publication No. 0337714; 0346847; 356223; 352000; 357332; 36,062; And 361341; Bone et al., IACS 113, 9382 (19910) However, all of these known HIV protease inhibitors are reversible inhibitors with increased affinity for enzymes, which are insufficient for use in actual therapeutics.

Accordingly, the present inventors have developed an irreversible inhibitor incorporating a cis-epoxide group unlike the transition state in order to obtain a stronger inhibitory effect than the reversible inhibitor that mimics the above-described transition state (by Applicant 1993). Korean Patent Application No. 93-10811, filed June 14, 2013), the present invention introduces an alkylamine carbamate to the (3R, 2S) cis-epoxide alcohol structure and the non-natural amino acid at the N-terminus. The introduction of sulfone-containing amino acids has led to the development of new compounds with stronger HIV proliferative inhibitory effects.

Accordingly, it is an object of the present invention to provide novel compounds with improved human immunodeficiency virus inhibitory activity and methods for their preparation.

Another object of the present invention is to provide a composition useful for the prevention or treatment of acquired immunodeficiency syndrome or HIV infection, comprising the compound as an active ingredient.

According to the present invention there is provided a compound of formula (I) having the cis-epoxide structure and pharmaceutically acceptable salts, hydrates or solvates thereof;

Where

R ₁ is selected from lower alkyl, lower alkyl substituted with an aromatic radical, and cyclic alkyl having 3 to 8 carbon atoms;

A is a functionalized acyl of the general formula (IIa) or (IIb) below;

(Wherein

R ₂ is selected from lower alkyl and lower alkyl substituted with amide,

R ₃ is selected from hydrogen and lower alkyl,

R ₄ is selected from hydrogen, lower alkyl, —NH ₂ and —OH,

1 is 0 or 1,

n is 0, 1 or 2),

B is a group of the following general formula (III);

R ₅ (Y) _m X (III)

(Wherein

X is -OC-, -SO-, -SO ₂ -or CS,

Y is -O-, -NH- or -NCH _3- ,

R ₅ is hydrogen, lower alkyl, cyclic alkyl, aryl, arylalkyl, aryloxy alkyl, aminoalkyl, alkylaminoalkyl, cyclicaminoalkyl, alkoxyalkyl, polyalkoxyalkyl, heterocycle, heterocyclicalkyl, arylthio Alkyl, arylsulfonylalkyl, heterocyclicthioalkyl, heterocyclicsulfonylalkyl, cycloalkoxyalkyl, cycloalkylthioalkyl, heterocyclicoxyalkyl and cycloalkylsulfonyl alkyl, m is 0 or 1),

D is a group of the following general formula (IV);

In which Z is O, NH or -NCH _3- ,

R ₆ and R ₇ are each independently selected from lower alkyl, lower alkyl substituted by aromatic radicals, cyclic radicals having 3 to 8 carbon atoms, lower alkyl substituted by cyclic radicals and aromatic radicals).

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

As used herein, the term “heterocycle” refers to a tri- or tetragonal compound comprising oxygen, nitrogen or sulfur atoms, a 5- or hexagonal compound comprising 1-3 heteroatoms selected from oxygen and sulfur atoms, In the case of pentagonal compounds, 0 to 2 double bonds are present, and in the case of hexagonal compounds, 0 to 3 double bonds are present. The nitrogen or sulfur atom may optionally be in oxidized form.

Heterocycles also include bicycles in which the aforementioned heterocycles are combined with benzene, cyclo hexane or other heterocycles. Representative heterocycles include pyrrole, pyrrolinyl, pyrrolidinyl, pyrazoyl, pyrazolinyl, pyrazolidinyl, imidazoline, imidazolinyl, imidazolidinyl, pyridyl, piperidinyl, pyrazinyl, pi Ferrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, isoxazolyl, isoxazolinyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl , Isothiazolidinyl, indolyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, benzimidazolyl, benzothiazoylyl, benzoxazoyl, benzopefranyl, furyl, dihydrofuranyl , Tetrahydrofuranyl, pyranyl, dihydro pyranyl, tetrahydro pyranyl, dioxanyl, dioxolanyl, thienyl, benzotianyl, benzotyranyl and the like.

The abbreviations for amino acids used in this specification are in accordance with the IUPAC-IUB consensus on biochemical nomenclature for amino acids and peptides. [Eur. J. Biochem., 158, 9-31 (1984).

The compounds according to the invention may also have asymmetric carbon centers and may exist as racemates, racemic compounds, diastereomeric mixtures and individual diastereomers, all of which are in the form of isomers.

Preferred compounds of the invention are those of formula (I) wherein R ₅ is selected from the group consisting of cyclicalkyl, arylalkyl, aryloxyalkyl, heterocycle, heterocyclicalkyl, heterocyclicthioalkyl and heterocyclicoxyalkyl Compound, more preferably,

R ₁ is isobutyl, cyclopentylmethyl, cyclohexylmethyl or benzyl,

R ₂ is a residue of an amino acid,

R ₃ is hydrogen, methyl or ethyl,

R ₄ is hydrogen, methyl or ethyl,

R ₅ is cyclohexylmethyl, benzyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, indolyl, pyridyl, pyridylmethyl, isoquinolinyloxymethyl, acetoxymethyl, tetrahydropyranyl, Benzopyranyl, thiazolylmethyl or thiazolyl,

R ₆ and R ₇ are each independently isopropyl, isobutyl, isopentyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclopropyl methyl or phenyl

It is a compound of general formula (I).

Representative examples of preferred classes of compounds according to the invention are as follows:

Compounds of formula (I) according to the invention can be prepared as shown in Scheme 1 below.

Scheme 1

Wherein A, B, D, and R ₁ to R ₄ are as defined above.

According to Scheme 1, the compound of Formula a is oxidized with MCPBA (metachloroperoxybenzoic acid), followed by coupling with the compound of Formula b to obtain a compound of Formula c, and then benzyloxycarbo The Nyl protecting group is removed to afford the compound of formula d. Subsequently, the compound d is subjected to a coupling reaction with the compound of the general formula e and then oxidized as described above to obtain a compound of the general formula f, and then the benzyloxycarbonyl protecting group is removed to remove the compound of the general formula g. This compound f is obtained by coupling with the compound of the general formula h or i to obtain the compound of the general formula (I).

The coupling reaction includes, for example, dicyclohexyl carbodiimide (DCC), 3-ethyl-3 '-(dimethylamino) -propylcarbodiimide (EDC), bis- (2-oxo-) as a coupling reagent. 3-oxazolidinyl) -phosphinic chloride (BOP-C1), diphenylphosphoryl azide (DPPA) and the like can be used, but are not limited thereto. Alternatively, the carboxylic acid of formula a may be converted into a halide or other active ester derivative and then subjected to a coupling reaction. The acid halide derivatives include acid chlorides, and active ester derivatives activate carboxylic acid groups to form amide bonds by coupling reactions with amines or to form ester bonds by coupling reactions with alcohols. Commonly used ones include, for example, alkoxy carbonyl halides such as methoxycarbonyl chloride, isobutoxycarbonyl chloride, and the like, carboxylic anhydrides derived from coupling reagents, and N-hydroxybenzotriazole derived esters. , N-hydroxyphthalimide derived esters-hydroxysuccinimide derived esters, N-hydroxy-5-norbornene-2′3′-dicarboxyamide derived esters, 2,4,5- Trichlorophenol derived esters and the like, but are not limited thereto.

The benzyloxycarbonyl leaving reaction can also be removed by conventional methods, for example by reaction under hydrogen pressure in the presence of a Pd / C catalyst.

On the other hand, the compound of Formula a may be prepared as shown in Scheme 2 below.

Scheme 2

Wherein X is a halogen atom, for example Br or I.

Scheme 2 illustrates a method for preparing cis-en-allylic alcohol, as described in Hiroshi et al., J. Chem, Soc., Chem. Commu., 311-312 (1987); and Adolphc Bohnstedt et al., Tetrahedron Letters, 34 (33), 5217-5220 (1993), analogously to the LN-benzyloxy carbonyl-R ₃ substitutions. The cis-en-allylic alcohol of the general formula a can be obtained by reacting the aldehyde with a digig.

Functionalized amines which provide a group of formula (IV) in the present invention can be prepared as shown in Schemes (3) and (4) below.

Scheme 3

tBOC: t-butoxycarbonyl

Scheme 4

Cbz: benzyloxycarbonyl

Wherein R ₆ , R ₇ and X are as defined above.

Scheme 3 is a process for preparing functionalized amines which provide the desired group of general formula (IV) by Grignard reaction and reduction reaction using N _a BH ₄ from nitrile (R ₆ CN). Since this product is formed into a racemate, the product is formed into a racemate, and after coupling with t-butoxycarbonyl-phenylalanine, the t-butoxycarbonyl group, which is a protecting group, is removed and subjected to column chromatography to separate into two diastereomers. Each isomer is then obtained by the Edman method.

Scheme 4 is a diagram showing a method for producing optically pure, wherein R ₆ is immobilized to an isopropyl group in a functionalized amine providing a group of general formula (IV), protected with N-benzyloxycarbonyl. The L- or D- amino acid of formula n with methoxymethyl amine, followed by Grignard reaction to give a compound of formula o. Subsequently, to prevent racemization, potassium hexamethyldisilazane (KHMDS) was used to carry out a Vitig reaction at −20 ° C. to obtain a compound of Formula q, and then hydrogenated to obtain the amine represented by Formula t. Can be synthesized.

The compound of general formula e used in Scheme 1 may be prepared as in Scheme 5 below.

Scheme 5

In this instance, R ₃ is as defined above, R ₄ is lower alkyl and X is a halogen atom, for example Br or I.

Scheme 5 shows a process for protecting the functionalized amine with a protecting group, wherein the thiol group of the substituted cysteine may be alkylated under a base and then protected with the benzyloxycarbonyl group.

Since the compound of the present invention has HIV protease inhibitory activity, it can be used for the purpose of treating or preventing AIDS or HIV infection. The total daily dose given to the host for this purpose is 100 to 600 mg / kg body weight, which may be administered once or several times, and the specific dose level for a particular patient will depend on the specific compound to be used, weight, age, sex and health. It may vary depending on the condition, diet, time of administration, method of administration, rate of excretion, drug mixing and the severity of the disease.

The compounds of the present invention can be parenterally or orally administered as desired. Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be prepared using suitable dispersing, wetting or suspending agents according to known techniques. Solvents that may be used include polyethylene glycol, ethylene glycol, polypropylene glycol, ethanol and the like. Solid dosage forms for oral administration may be capsules, tablets, topical agents, powders and granules. However, in view of compound properties, capsules are particularly preferred, and tablets may be useful in preparation of enteric preparations. Solid dosage forms may contain the active compound in one or more inert diluents such as sucrose, lactose or starch and lubricants such as magnesium stearate in addition to the diluent.

The compounds of the present invention may also be administered in parallel with one or more other anti-AIDS agents, immunomodulators and the like. 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 useful agent of the compounds of the present invention for the treatment and prevention of HIV infection may be included.

Such a present invention will be described in more detail based on Production Examples and Examples. The following examples are only for the understanding of the preparation method of the novel compounds according to the present invention, but do not limit the scope of the present invention.

Preparation Example 1 Preparation of N-benzyloxycarbonyl-β- (S-methyl) -L-valine

8.9 g (0.06 mol) of β-mercapto-L-valine was added to a mixture of 120 ml of dioxane and 40 ml of water, cooled to 0 ° C., and then dissolved by adding 20 ml of 6N aqueous sodium hydroxide solution. 9.24 g (0.066 mol) of iodine methane was added to the solution, and the flask was capped and then reacted at 0 ° C. for 3 hours and then at room temperature for 2 hours. The methylation reaction was cooled to 0 ° C. and 15 mL of 6N aqueous sodium hydroxide solution and 10.20 g (0.09 mol) of benzylchloroformate were slowly added. The reaction was stirred at 0 ° C. for 1 hour and then at 5 ° C. for 2 hours to terminate the reaction. After completion of the reaction, the solvent was distilled off under reduced pressure, and then 20 mL of water and ether were added to remove unreacted benzylchloroformate, and then the organic layer was removed. 60 ml of ethyl acetate was added to the aqueous layer and the pH was lowered to 3 or less with 6N hydrochloric acid. The organic layer was separated, dried over anhydrous MgSO ₄ , and the solvent was evaporated under reduced pressure to yield 14.25 g (yield 80%) of the title compound.

¹ H NMR (CD ₃ OD) δ 1.2 (s, 6H), 2.1 (s, 3H), 4.3 (d, 1H), 5.1 (s, 2H), 7.1 (m, 5H)

Preparation Example 2

Step 1: Preparation of 2-methyl-3-R-benzyloxycarbonylamino-4-phenyl-1-butene

After dissolving 5.7 g (0.012 mol) of methyltriphenyl phosphine bromide in 40 ml of anhydrous toluene, it was cooled to -20 DEG C, and 22 ml (0.011 mol) of 0.5N potassium hexamethyl disilazine solution was slowly added under a nitrogen atmosphere. It was. Then, 2.63 g (0.01 mol) of R-3-benzyloxycarbonylamino-4-phenyl-2-butanone synthesized by the method of Nahm (Tetrahedron Letter, 54, 3815 (1981)) was slowly added. . After reacting at the same temperature for 30 minutes, the temperature of the reactant was gradually raised to room temperature, followed by stirring for 3 hours. After completion of the reaction, the solvent was removed by distillation under reduced pressure and column chromatography (eluent; ethyl acetate: hexane = 10: 90) was performed to obtain 2.2 g (84%) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 1.77 (s, 3H), 2.65-2.95 (m, 2H), 4.27 (br, 1H), 4.51 (b, 1H), 4.80 (d, 2H), 5.15 (s, 2H ), 7.19-7.33 (m, 10H)

Step 2: Preparation of (R) -2-amino-3-methyl-1-phenyl butane

100 mg of 10% palladium on carbon was added to a solution obtained by dissolving 2-methyl-3-R-benzyloxycapnylamino-4-phenyl-1-butene in 2.61 g (0.01 mol) of 30 ml of methanol, followed by 1 The mixture was stirred for 3 hours under atmospheric pressure hydrogen conditions (rubber balloon). The reaction solution was passed through celite to remove the inorganic metal, and then the reaction solvent was removed to obtain 1.43 g (yield 87%) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 0.98 (t, 6H), 1.65 (m, 1H), 2.39 (m, 1H), 2.82 (m, 2H), 7.19-7.33 (m, 5H)

Preparation Example 3

Step 1: Preparation of 2-amino-3-methyl-1-phenylbutane hydrochloride

To 13.8 g (0.2 mol) of isobutylnitrile diluted with 50 ml of anhydrous THF (tetrahydrofuran) was added 110 ml (0.22 mol) of 2.0 M benzyl magnesium chloride at room temperature, refluxed for 1 hour and then cooled to room temperature.

Then, 200 ml of methanol and 11.4 g (0.3 mol) of NaBH ₄ were added, followed by stirring at room temperature for 1 hour. 1N hydrochloric acid was added to adjust the pH to about 11. The mixture was extracted with chloroform, dried over anhydrous Na ₂ SO _4, and methanolic hydrochloric acid was added to give 37.5 g (94% yield) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 1.08 (m, 6H), 1.96 (m, 1H), 2.90-3.16 (m, 2H), 3.37 (m, 1H), 7.15-7.31 (m, 5H), 8.36 (b , 3H)

Step 2: Preparation of L- (N-t-butoxycarbonyl) -phenylalaninyl-2- (1-phenyl-3-methylbutyl) amide

1.5 equivalents of EDC and HOBT were added to 26.5 g (0.1 mol) of Nt-butoxycarbonyl phenylalanine, respectively, and then dissolved by adding 130 ml of DMF and 15 ml of triethylamine. To the resulting solution, 20 g (0.1 mole) dmf of the product of step 1 was added at 0 ° C., and stirred at room temperature for 5 hours. The solvent was then removed by distillation under reduced pressure, dissolved in ethyl acetate, washed with 1N hydrochloric acid and saturated solution of NaHCO ₃ , the organic layer was dried over anhydrous MgSO _4, and the solvent was removed to give 37.7 g (92% yield) of the title compound. Got it.

Step 3: Preparation of L-phenylalaninyl-2- (1-phenyl-3-methylbutyl) amide

20.5 g (0.05 mol) of the product of step 2 was dissolved in 30 ml of dichloromethane and 15 ml of trifluoroacetic acid, stirred at room temperature for 1 hour, the solvent was distilled off under reduced pressure, and column chromatography was performed using ethyl acetate. Two isomers were separated by running. As a result, 8.0 g of isomers having an Rf value of 0.50 and 7.4 g having an Rf value of 0.45 were obtained. Overall yield was 99%.

¹ H NMR (CDC1 ₃ ) δ

1.Rf = 0.50: 0.93 (m, 6H), 1.80 (m, 1H), 2.22 (m, 1H), 2.63 (m, 1H), 2.85 (m, 1H), 3.05 (m, 1H), 3.51 ( m, 1H), 4.11 ((m, 1H), 7.10-7.34 (m, 10H)

2.Rf = 0.45: 0.91 (m, 6H), 1.79 (m, 1H), 2.65-2.70 (m, 2H), 2.83 (m, 1H), 3.18 (m, 1H), 3.44 (m, 1H), 4.08 (m, 1 H), 7.10-7.32 (m, 10 H)

Step 4: Preparation of 2-amino-3-methyl-1-phenylbutane

1.46 g (4.7 mmol) of each isomer of step 3 was dissolved in 50 mL of anhydrous dichloromethane. To the resulting solution, 0.66 mL (5.5 mmol) of phenylisothiocyanate was added at room temperature, refluxed for 2 hours, and then cooled to room temperature. 10 ml of fluoroacetic acid was added thereto, refluxed at 60 DEG C for 40 minutes, and then the solvent was distilled off. The product was then dissolved in 20 mL of water, washed with ether, adjusted to pH 11 with NaOH and extracted with chloroform to give each of the isomers of the title compound in a yield of 82-85%.

¹ H NMR (CDC1 ₃ ) δ 0.94 (m, 6H), 1.11 (bs, 2H), 1.65 (m, 1H), 2.39 (m, 1H), 2.82 (m, 2H), 7.16-7.32 (m, 5H )

[α] _D = 1. -38.1 (c = 0.12, dichloromethane)

2. +38.1 (c = 0.12, dichloromethane)

Preparation Example 4

Step 1: Preparation of [1- [4S- (N-benzyloxycarbonyl) amino] -5-phenyl-cis-2-pentene]-[tetrahydropyran] ether

5.64 g (12 mmol) of 2- (2-bromoethyloxy) tetrahydropyranylether triphenylphosphine salt were dissolved in 40 ml of tetrahydrofuran and stirred at -78 ° C. 25 ml of potassium hexamethyldisilazane 0.5M solution was added and maintained at -78 ° C for 1 hour. 3.0 g (10.6 mmol) of L- (N-benzyloxycarbonyl) phenylalanineal were then dissolved in 15 ml of tetrahydrofuran and added slowly to the solution at -78 ° C and at -78 ° C for 1 hour and then at room temperature. After stirring for 1 hour, water was added to terminate the reaction. The reaction solvent was then removed and dissolved in ethyl acetate, the resulting solution was washed with saturated NaHCO ₃ solution and water, and then the organic layer was dried over anhydrous MgSO ₄ and subjected to column chromatography to give 1.96 g (45% yield) of the title. The compound was obtained.

¹ H NMR (CDC1 ₃ ) δ 1.81 (m, 6H), 3.01 (m, 2H), 3.63 (m, 1H), 4.21 (m, 3H), 4.6-4.9 (br, 3H), 5.13 (s, 2H ), 5.41 (t, 1H), 5.62 (m, 1H) 7.0-7.4 (m, 10H)

Step 2: Preparation of 4S (N-benzyloxycarbonyl) amino-5-phenyl-cis-2-penten-1-ol 2.47 g (6 mmol) of the compound obtained in step 1 was dissolved in 80 ml of ethanol, 0.2 equivalents of PPTS (pyridinium para-toluenesulfonate) was added dropwise and stirred at room temperature for 24 hours. Subsequently, the organic solvent was removed by distillation under reduced pressure, and then dissolved in dichloromethane solvent, washed with saturated NaHCO ₃ solution, and the organic layer was dried over anhydrous MgSO ₄ to remove the solvent, followed by column chromatography (eluent; ethyl acetate: hexane = 2: 8) to yield 1.67 g (90% yield) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 2.82 (m, 2H), 3.41 (m, 3H), 4.32 (m, 1H), 4.83 (br, 1H), 5.01 (s, 2H), 5.21 (t, 1H), 5.42 (m, 1 H) 7.0-7.4 (m, 10 H)

Step 3: Preparation of 4S- (N-benzyloxycarbonyl) amino-5-phenyl- (3R, 2S) -epoxypentan-1-ol alcohol

311 mg (1 mmol) of the product of step 2 was dissolved in 20 ml of dichloromethane, 3 equivalents of metachloroperoxy benzoic acid was added, and stirred at room temperature for 24 hours. 30 ml of concentrated Na ₂ S ₂ O ₃ solution was added thereto, stirred for 30 minutes, and the organic layer was washed with saturated NaHCO ₃ solution.

The organic layer was then dried over anhydrous MgSO ₄ and the organic solvent was removed to give 198 mg (63% yield) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 2.8-3.2 (m, 6H), 3.81 (m, 1H), 4.83 (br, 1H), 5.01 (s, 2H), 7.0-7.4 (m, 10H)

Example 1: [4S-[[N- (2-tunolincarbonyl) amino] -L-asparazinyl] amino]-(3R, 2S) -epoxy-5-phenyl-pentoxy]-[(2R Preparation of -amino-1-phenyl-3-methyl] butyl] carbamate

Step: 1 [(4S)-[N-benzyloxycarbonyl) amino]-(3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(2R-amino-1-phenyl-3-methyl ] Butyl] carbamate

198 mg (0.6 mmol) of the product obtained in step 3 of Preparation Example 4

[4s-[(N-benzyloxycarbonyl) amino]-(3R, 2S) -epoxy, synthesized according to the method of Argo, Ghosh et el., Tetrahedron Letter 33, 2781-2784 (1992). 122 mg (0.7 mmol) of the product of Preparation Example 2, Step 2 and 3 equivalents of triethylamine in -5-phenyl-1-pentoxy] succinimidylcarbonate were dissolved in 30 ml of a dichloromethane solution at room temperature. I ate for 2 hours. Subsequently, the resultant was washed with saturated NaHCO ₃ solution, and the organics were dried over anhydrous MgSO ₄ to remove the organic solvent, and then subjected to column chromatography (ethyl acetate: hexane = 3: 7) to give 258 mg (yield 81%) of the title compound. .

¹ H NMR (CDC1 ₃ ) δ 0.81 (dd, 6H), 2.02 (m, 1H), 2.8-3.1 (m, 6H), 3.6-4.0 (m, 4H), 4.9-5.3 (m, 4H), 7.0 -7.4 (m, 15H)

Step 2: [(4S-[[N- (2-Quinolinecarbonyl) amino] -L-asparazinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(2R Preparation of -amino-1-phenyl-3-methyl) butyl] carbamate

258 mg (0.5 mmol) of the product of step 1 was dissolved in 50 ml of methanol, and then 10% by weight of 10% Pd / C was mixed and stirred for 3 hours under hydrogen (balloon) conditions. The reaction solution was passed through celite to remove the inorganic catalyst, and then the reaction solvent was removed by distillation under reduced pressure. 173 mg (0.6 mmol) of N- (2-quinolincarbonyl) asparagine, 1.5 equivalents of EDC and HOBT, respectively, were dissolved in 10 ml of dimethylformamide, followed by addition of the amine obtained above at 0 ° C and 6 hours at room temperature. Was complained about. The solvent was removed by distillation under reduced pressure, dissolved in dichloromethane, and washed with saturated NaHCO ₃ solution. The organic layer was dried over anhydrous Na ₂ SO _4, and the organic solvent was distilled off under reduced pressure, followed by column chromatography (pec acid: ethyl acetate = 3: 7) to obtain 140 mg (yield 43%) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 0.81 (dd, 6H), 2.01 (m, 1H), 2.6-3.0 (m, 8H), 3.6-4.4 (m, 5H), 5.02 (m, 1H), 5.41 (br , 2H), 6.43 (br, 1H), 7.0-7.4 (m, 10H), 7.5-8.3 (m, 7H), 9.43 (br, 1H)

FAB MS (M + 1) 652

Examples 2 and 3

The reaction was carried out in a similar manner as in Example 1 to obtain the compounds listed in Table 1 below.

Example 4 [4S)-[(N-benzyloxycarbonyl) amino-β-methanesulfonyl-L-valinyl] amino]-(3R, 2S) -epoxy-5-phenyl-1-pentoxy] Preparation of-[(2R-amino-1-phenyl-3-methyl) butyl) carbamate

258 mg (0.5 mmol) of the product of Example 1, Step 1 was dissolved in 50 ml of methanol, and then 10% by weight of 10% Pd / C was mixed and stirred for 3 hours under hydrogen (balloon) conditions. The reaction solution was passed through celite to remove the inorganic catalyst, and then the reaction solvent was removed by distillation under reduced pressure. 148 mg (0.5 mmol) of the title compound of Preparation Example 1, 1.5 equivalents of EDC, HOBT, and triethylamine, respectively, were dissolved in 50 ml of dimethylformamide, and the obtained amine was added at 0 ° C. for 3 hours at room temperature. Stirred. The solvent was removed by distillation under reduced pressure, dissolved in dichloromethane, and washed with saturated NaHCO ₃ solution. The organic layer was dried over anhydrous MgSO ₄ and 5 equivalents of metachloroperoxybenzoic acid was added and stirred at room temperature for 2 hours. A 10% Na ₂ S ₂ O ₃ solution was added, stirred for 30 minutes, and the organic layer was washed with saturated NaHCO ₃ solution. The organic layer was dried over anhydrous MgSO _4, and the organic solvent was removed, followed by column chromatography (hexane: ethyl acetate = 3.7) to give 263 mg (yield 76%) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 0.82 (d, 6H), 1.4-2.0 (m, 7H), 2.6-3.9 (m, 13H), 4.63 (br, 1H), 4.81 (d, 1H), 5.0 (s , 2H), 5.82 (br, 1H), 7.0-7.5 (m, 11H)

FAB MS (M + 1) 694

Examples 5 and 6

The reaction was carried out in a similar manner as in Example 4, to obtain the compounds listed in Table 1 below.

Example 7 [4S-[[N- (2-Quinolinecarbonyl)] amino-β-methanesulfonyl-L-valinyl] amino]-(3R, 2S) -epoxy-5-phenyl-1-phene Preparation of oxy]-[(2R-amino-1-phenyl-3-methyl) butyl) carbamate

347 mg (0.5 mmol) of the product of Example 4 was dissolved in 20 ml of methanol, and then 10% by weight of 10% Pd / C was mixed and stirred for 3 hours under hydrogen (balloon) conditions. The reaction solution was passed through celite to remove the inorganic catalyst, and then the reaction solvent was removed by distillation under reduced pressure. 86.6 mg (0.5 mmol) of N- (2-quinoline carbonyl acid) and 1.5 equivalents of EDC and HOBT, respectively, were dissolved in 10 ml of dimethylformamide, and the amine obtained above was added at 0 ° C. for 6 hours at room temperature. Stirred. The solvent was removed by distillation under reduced pressure, dissolved in dichloromethane, and washed with saturated NaHCO ₃ solution. The organic layer was dried over anhydrous Na ₂ SO _4, and the organic solvent was distilled off under reduced pressure, and then subjected to column chromatography (hexane: ethyl acetate = 3: 7) to give 221 mg (yield 62%) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 0.81 (m, 6H), 1.4-2.0 (m, 7H), 2.6-3.8 (m, 12H), 4.12 (m, 2H), 4.63 (br, 1H), 4.82 (d , 1H), 5.42 (br, 1H), 7.0-7.5 (m, 10H), 9.33 (br, 1H)

FAB MS (M + 1) 715

Examples 8 and 9

The reaction was carried out in a similar manner as in Example 7, to obtain the compounds listed in Table 1 below.

Example 10 [4S-[[[N- (5-isoquinolyloxy)] methylenecarbonyl] amino-β-methanesulfonyl-L-valinyl] amino]-(3R, 2S) -epoxy-5 Preparation of -phenyl-1-pentoxy]-[(2R-amino-1-phenyl-3-methyl) butyl) carbamate

347 mg (0.5 mmol) of the product of Example 4 was dissolved in 20 ml of methanol, and then 10% by weight of 10% Pd / C was mixed and stirred under hydrogen (balloon) conditions for 3 hours. The reaction solution was passed through celite to remove the inorganic catalyst, and then the reaction solvent was removed by distillation under reduced pressure. Dissolve 101 mg (0.5 mmol) of 5-isoquinolinyloxy acetic acid and 1.5 equivalents of EDC and HOBT in 10 ml of dimethylformamide, then add the amine obtained above at 0 ° C. and stir at room temperature for 3 hours. It was. The solvent was removed by distillation under reduced pressure, dissolved in dichloromethane, and washed with saturated NaHCO ₃ solution. The organic layer was dried over anhydrous Na ₂ SO _4, and the organic solvent was distilled off under reduced pressure, and then subjected to column chromatography (hexane: ethyl acetate = 3: 7) to give 316 mg (yield 85%) of the title compound.

¹ H NMR (CDC1 ₃ ) δ 0.83 (m, 6H), 1.5-2.0 (m, 7H), 2.4-3.8 (m, 12H), 4.13 (m, 12H), 4.13 (m, 1H), 4.63 (br , 1H), 4.71 (s, 2H), 4.82 (d, 1H), 5.82 (br, 1H), 7.0-8.5 (m, 16H), 9.32 (br, 1H)

FAB MS (M + 1) 745

Examples 11 and 12

The reaction was carried out in a similar manner as in Example 10, to obtain the compounds listed in Table 1 below.

Table 1

Inhibitory Effect of HIV Protease

The following method was used to confirm the inhibitory efficacy of the compounds of the present invention against HIV proteases.

First, the enzyme is mixed with the compound prepared in Examples 1 to 12 of the present invention in the absence of the reactor (Bangyang solution), and then a portion of the reaction solution is taken and added to the solution having an excess of the reactor (Analysis solution). The degree of decrease in enzyme activity was measured as the degree of better activity. The pre-culture solution is 50 mM sodium acetate, pH 0.5, 1 mM dithiothreitol (DTT), 1 mM ethyl diamine tetraacetate (EDTA), 0.75 M ammonium sulfate, 0.1% Np 40 and various concentrations of the compounds of Examples 1-12. This is included. The inhibitory response was initiated by adding 2.6 nM HIV-1 protease. Every 10 hours the solution was added to 10 μl of solution to assay for remaining enzyme activity. At this time, the oligopeptide consisting of 11 amino acids of H-His-Lys-Ala-Arg-Val-Leu-Phe- (p-nitro)-(Glu-Ala-Ile-Ser-NH ₂ ) was used as the reactor. The substrate breaks the amide bond between (p-nitro) -Phe and Leu by HIV protease. The reaction rate was determined by using a strong hop luminous intensity at (280 nm) of (p-nitro) -Phe to separate the substrate before the reaction and the product after the reaction by HPLC to measure the relative amounts of the product. K _obs was calculated from the slope of the graph by calculating the decrease in enzymatic activity with time and plotting the natural log value (1n) of the decrease over time. Inhibition constant was calculated | required by the following formula.

The above equation can be adapted to the case where the inhibitor concentration is much higher than the enzyme concentration (steady-state conditions). However, if the concentration of inhibitor is high and the concentration of inhibitor and enzyme is not sufficient, the formula E + I

(E: free enzyme concentration, I: free inhibitor concentration, EI: Michael-Menten complex concentration, EI ': covalent conjugate concentration of enzyme and inhibitor) The inhibitory constants K _I and K _ina and the second order constants are calculated by calculating the relative concentration of enzyme [E] / ([E] + [EI] + [EI ']) by entering them into the KINSIM / FITSIM program. K _ina / K _I (Constant) was obtained. The results of the inhibition constants are shown in Table 2 below.

In the case of the compound of the present invention, since the value of the second inhibitory constant (k _ina / K _I ) is sufficiently large, it can be seen that an irreversible reaction occurred between the enzyme and the inhibitor.

Antiviral Activity and Cytotoxicity Measurement

Antiviral effect was determined by the concentration of 50% inhibiting viral replication (IC ₅₀ ) by the study of C. chitin formation and reverse transcriptase assay.

H9 cell line and Sup T1 cell line of 1 × 10 ⁵ cells were placed in a 24-well plate and various concentrations of the compound of the present invention were added. HIV-1 inoculum was added to 200TCID ₅₀ (200-fold 50% cell infection concentration, Tissue Culture Infectious Dose) and Rpmi-1640 medium was added and cultured at 37 ° C. In the case of Sup T1, the number of synchitium was formed after 3 to 9 days. In the case of H9, three quarters of the culture solution was changed to a new culture solution of the same concentration every three days. After 9 days, 6 ml of the culture was taken, 2.5 ml of 30% PEG (polyethylene glycol, molecular weight about 6000 to about 8000) and 0.4 M NaC1 were added, and the mixture was left overnight at 0 ° C. to precipitate the virus. Centrifuge at 2000 rpm for 45 minutes to discard the supernatant, dilute the precipitate with 20 µl of reverse transcriptase suspension, and place it in an Eppendorf tube. Store at -70 ° C until use. The suspension buffer composition contains 50 mM Tris buffer (Sigma), pH 7.5, 1 mM dithiothreitol, 20% glycerol, 0.25M KC1 and 0.25% Triton X-100. The suspension was frozen in dry ice for 2 minutes and then dissolved three times at 37 ° C. for three minutes, followed by centrifugation at 4 ° C., and reverse transcriptase assay using a supernatant. Reverse transcriptase assay solution contains 10 μl of the virus suspension, 10 μl Tris buffer (250 mM Tris, pH 7.5, 37.5 mM MgC1 ₂ , 0.25% Triton X-100), 1.2 μl 200 mM dithiothreitol, 5 μl 100 μM (dT) -poly (A) (Boeringer Mannheim, 12-18 oligomers), 1 μl (1 μCi) of TTP (thymine triphosphate) containing ³ H, and 23.6 μl of water were used in combination. After time, the solutions are poured into Whatman DEB1 filter paper and then placed in 2 X SSC buffer. Wash three times with 2 X SSC buffer (taken for about 10 minutes each time), then wash twice with 10% of 95% methanol. Filter paper is placed in aluminum foil, dried with an infrared bulb and quantified with a liquid scintillation counter.

To test the cytotoxicity to determine the maximum allowance of anti-AIDS agent, the compounds in the range of 0.1 μM to 100 μM were added to H9 cells or Sup T1 cells, and then incubated at 37 ° C. in Rpmi-1640 medium and cultured at 3 days intervals. The cytotoxicity CT ₅₀ was determined after observing the proliferation of the cells for 2 weeks in a hemocytometer plate by Trypan blue dye exclusion technique. The results of the measured antibacterial activity and cytotoxicity are shown in Table 2 below.

TABLE 2

* If k _ina / K _I is more than 10 ⁹ , there is a limit to get accurate value by KINSIM / FITSIM. Willams, JW and Morrison, JF, Methods, Enzymo 1., 63, 437-466 (1079); and Zimmerle, CT and Frieden, C., J. Biochem., 258, 381-387 (1989).

As described above, the compound of the general formula (I) of the present invention is an irreversible inhibitor of HIV protease, which has high inhibitory effect and low cytotoxicity, and thus is useful as a pharmaceutical composition for treating or preventing HIV or HIV infection. Can be used.

Claims (5)

A compound of formula (I) having the cis-epoxide structure or a pharmaceutically acceptable salt, hydrate or solvate thereof: Where R ₁ is selected from lower alkyl, lower alkyl substituted with an aromatic radical, and cyclic alkyl having 3 to 8 carbon atoms; A is a functionalized acyl of the general formula (IIa) or (IIb) below; (Wherein R ₂ is selected from lower alkyl and lower alkyl substituted with amide, R ₃ is selected from hydrogen and lower alkyl, R ₄ is selected from hydrogen, lower alkyl, —NH ₂ and —OH, 1 is 0 or 1, n is 0, 1 or 2), B is a group of the following general formula (III); R ₅ (Y) _m X (III) (Wherein X is -OC-, -SO-, -SO ₂ -or CS, Y is -O-, -NH- or -NCH _3- , R ₅ is selected from the group consisting of aryl, arylalkyl, heterocycle and heterocyclylalkyl, m is 0 or 1). D is a group of the following general formula (IV); (Wherein Z is 0, NH or -NCH _3- , R ₆ and R ₇ are each independently selected from lower alkyl, lower alkyl substituted by aromatic radicals, cyclic radicals having 3 to 8 carbon atoms, lower alkyl substituted by cyclic radicals and aromatic radicals). The method of claim 1, R ₁ is isobutyl, cyclopentylmethyl, cyclohexylmethyl or benzyl, R ₂ is a residue of an amino acid, R ₃ is hydrogen, methyl or ethyl, R ₄ is hydrogen, methyl or ethyl, R ₅ is benzyl, quinolinyl or isoquinolinyl, R ₆ and R ₇ are each independently isopropyl, isobutyl, isopentyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclopropyl methyl or phenyl Compound or a pharmaceutically acceptable salt, hydrate or solvate thereof. The method of claim 1, [4S-[[N- (2-Quinolinecarbonyl) -L-asparazinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(2R-amino-1-phenyl -3-methyl) butyl] carbamate, [4S-[[N- (2-Quinolinecarbonyl) amino] -L-asparazinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(1R-amino-1 -Phenyl-2-methyl) propyl] carbamate, [4S-[[N- (2-Quinolinecarbonyl) amino] -L-asparazinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(3-amino-2 , 4-dimethyl) pentyl] carbamate, [4S-[[N- (2-benzyloxycarbonyl) amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[( 1R-amino-1-phenyl-3-methyl) butyl] carbamate, [4S-[[N- (2-benzyloxycarbonyl) amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[( 1S-amino-1-phenyl-2-methyl) propyl] carbamate, [4S-[[N- (2-benzyloxycarbonyl) amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[( 3-amino-2,4-dimethyl) pentyl] carbamate, [(2R-amino-1-phenyl-3-methyl) butyl] carbamate, [4S-[[N- (2-benzyloxycarbonyl) amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[( 1S-amino-1-phenyl-2-methyl) propyl] carbamate, [(3-amino-2,4-dimethyl) pentyl] carbamate, [4S-[[N- (2-Quinolinecarbonyl) amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(2R -Amino-1-phenyl-3-methyl) butyl] carbamate, [4S-[[N- (2-Quinolinecarbonyl) amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(1S -Amino-1-phenyl-2-methyl) propyl] carbamate, [4S-[[N- (2-Quinolinecarbonyl) amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1-pentoxy]-[(3 -Amino-2,4-dimethyl) pentyl] carbamate, [4S-[[N- (5-isoquinolinyloxy)] methylenecarbonyl] amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1- Pentoxy]-[(2R-amino-1-phenyl-3-methyl) butyl] carbamate, [4S-[[N- (5-isoquinolinyloxy)] methylenecarbonyl] amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1- Pentoxy]-[(1S-amino-1-phenyl-2-methyl) propyl] carbamate, [4S-[[N- (2-isoquinolinyloxy)] methylenecarbonyl] amino-β-methanesulfonyl-L-valinyl] amino- (3R, 2S) -epoxy-5-phenyl-1- Pentoxy]-[(3-amino-2,4-dimethyl) pentyl] carbamate, and A compound selected from pharmaceutically acceptable salts, hydrates or solvates thereof. Oxidizing the compound of the general formula a followed by coupling reaction with a compound of the general formula b to obtain a compound of the general formula c; Removing benzyloxycarbonyl protecting group from compound c to obtain a compound of the general formula d; Compound d is subjected to a coupling reaction with a compound of the general formula e and then oxidized to remove the benzyloxycarbonyl protecting group from the following general formula f to obtain a compound of the general formula g; A process for preparing a compound of formula (I) according to claim 1 comprising coupling a compound g to a compound of formula h or i: Wherein A, B, D, and R ₁ to R ₄ are as defined in claim 1. A composition for the treatment or prevention of acquired immunodeficiency or human immunodeficiency virus infection comprising the compound according to claim 1 or a pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient.