WO1996018627A1

WO1996018627A1 - Ahpba-containing tripeptide derivative

Info

Publication number: WO1996018627A1
Application number: PCT/JP1995/002546
Authority: WO
Inventors: Yuichiro Yabe; Takashi Watanabe; Takashi Nishigaki; Yuji Ozawa; Tomoaki Komai; Akihiko Nakagawa
Original assignee: Sankyo Company, Limited
Priority date: 1994-12-14
Filing date: 1995-12-13
Publication date: 1996-06-20
Also published as: AU4188096A

Abstract

A novel AHPBA-containing tripeptide derivative represented by general formula (I), having an excellent HIV protease inhibitory activity, and being useful as a preventive or remedy for HIV infection, wherein R1 represents quinoline-2-carbonyl or quinoxaline-2-carbonyl; and R2 represents H, alkyl, acetyl, methoxymethyl, methylthiomethyl, acetoxymethyl, pivaloyloxymethyl, HOOC(CH¿2?)nCO- (wherein n represents an integer of 2 to 4), or HOOCCH2OCH2CO-.

Description

Specification

Technical Field The present invention relates to a novel human protease having an inhibitory activity on a human derived immunodeficiency syndrome onset virus (hereinafter referred to as HIV) -derived protease (hereinafter referred to as HIV protease). The present invention relates to a tripeptide derivative containing amino-2-hydroxy-4-phenylbutanoic acid (hereinafter, AHPBA). Background art

AIDS is a disease caused by a retrovirus called HIV, one of the lentiviruses. According to WHO estimates, about 1 million people worldwide are affected by the disease today, and the number of patients is ever increasing. The disease is fatal and there is currently no effective cure for cure.

A general feature of retrovirus replication is that the viral protease processes the viral precursor protein to produce the viral protein required for virus composition and function. Inhibiting this processing usually prevents the production of infectious virus. For example, Kohl, NE et al., Proc. Natl. Acad. Sci. 85, 4686 (1988), report that mature infectious virions when HIV-encoded proteases are genetically inactive. Has not been produced. This indicates that that force ⁵ to inhibit HIV protease Ichize ', prevention of HIV infection properly is effective in the treatment of treatment and AIDS Things.

Based on this belief, a number of HIV protease inhibitor trades have already been synthesized, and some substances found in nature and exhibiting anti-HIV activity at / v / ir have been reported [Lang., Rosel J. Arch. Pharm. 326, 921 (1993); Martin, JA, Antiviral Ros., 17, 265 (1992);

Meek, T.D., J. of Enzyme Inhibition, _6_, 65 (1992); JP-A-5-222020; J. Med. Chem. 1993, 36, 292)]].

However, in general, when these HIV protease inhibitors are administered in vivo or parenterally to a living body, it is difficult to maintain a blood concentration sufficient to suppress virus replication in infected cells in the body. In fact, few HIV protease inhibitors are in clinical trials.

The present invention includes a transition-state analog that is not cleaved by HIV protease, is specific for HIV protease, has excellent enzyme inhibitory activity, is effective at low levels on infected cells, and is useful in vivo. Provided is a low-molecular-weight HIV protease inhibitor that achieves a blood level that can be expected to be effective by oral or parenteral administration. DISCLOSURE OF THE INVENTION The novel AHPBA-containing peptide derivatives of the present invention

General formula

H / [Wherein, Rl represents a quinolin-12-carbonyl group or a quinoxaline-12-carbonyl group, R2 represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, an acetyl group, a methoxymethyl group, Represents a methylthiomethyl group, an acetyloxymethyl group, a bivaloyloxymethyl group, a HOOC (CH ₂ ) _n CO— group (n represents an integer of 2 to 4) or a HOOCCH ₂₀ CH ₂ CO— group And a salt thereof. BEST MODE FOR CARRYING OUT THE INVENTION In the above general formula (I):

Examples of the alkyl group having 1 to 4 carbon atoms of R ² include a methyl, ethyl, _n- propyl, isopropyl, n-butyl, isobutyl, s-butyl and t-butyl group. Is a methyl or t-butyl group.

The compound (I) of the present invention may be converted into a salt in some cases. Preferably, the compound (I) is an alkali metal salt such as a sodium salt or a potassium salt, or a calcium salt. Metal salts such as alkaline earth metal salts such as magnesium and magnesium salts; hydrohalides such as hydrochloride, hydrobromide and hydroiodide, nitrate, perchlorate and sulfuric acid Inorganic acid salts such as salts and phosphates; lower alkane sulfonates such as methanesulfonate, trifluormethanesulfonate and ethanesulfonate, benzenesulfonate, p_toluenesulfonate Organic acid salts such as arylsulfonate, fumarate, succinate, citrate, tartrate, oxalate, maleate, etc. and glutamate, aspartate Amino acid salts such as Can be When the compound (I) of the present invention is left in the air, it may absorb water, adhere to adsorbed water, or form hydrates. The compounds (I) of the present invention also include stereoisomers having an asymmetric carbon in the molecule and each having an R configuration or an S configuration, and each of them or a mixture thereof of Both are included in the present invention.

Among the compounds of the present invention, preferred are:

2) R2 is a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, an acetyl group, a methoxymethyl group, a HO OC (C H2) _η CO— group (n represents an integer of 2 to 4) or H 00 A compound which is a CC H2 0 CH ₂ CO— group,

3) compounds wherein R2 is a hydrogen atom or an alkyl group having 1 to 4 carbon atoms,

4) A compound in which R2 is a hydrogen atom.

Representative compounds of the present invention are shown in Table 1 below, but the compounds of the present invention are not limited thereto.

In Table 1, Me represents a methyl group, 丄 Bu represents a t-butyl group, M0M represents a methoxymethyl group, Ac represents an acetyl group, and Suc represents a succinyl group (H 00 C (CH 2) 2 CO—), Dig indicates a diglycolyl group (H00 CC H2 0 C H2 CO—), and Glt indicates a glutaryl group (H 00 C (C H2) 3 C-one). ), M SM represents a methylthiomethyl group, A OM represents an acetomethyl group, Q xa represents a quinoxaline 12-carbonyl group, and Q oa represents a quinolinyl 12-carbonyl group. And PM represents a pinoloyloxymethyl group.

(table 1 )

Example number R2

1 Q x a H 2 Q x a M e 3 Q x a 丄 B u 4 Q x a A c 5 Q x a MOM 6 Q x a S ue 7 Q x a D i g 8 Q x a G 1 t 9 Q x a SM

1 0 Q xa A〇M 1 1 Q xa P OM 1 2 Q oa H 1 3 Q oa M e 1 4 Q o.a 丄 B u 1 5 Q oa A c 1 6 Q oa MOM 1 7 Q oa S ue 1 8 Q oa D ig 1 9 Q oa G 1 t

2 0 Q o a M S M

2 1 Q o a A OM

2 2 Q o a P OM In the above table, preferred compounds are

2 (S), 3 (S) — 3 — [N-(quinoxaline-1-carbonyl)-1-L-threonyl] amino 2-hydroxy-1-phenylbutanoyl [

4 (S) — black mouth] — L-proline t e rt — butylamide (example number ュ) and

2 (S), 3 (S) -3-3- [N- (quinoline-l-carbonyl) -l-threonyl] amino 2-hydroxyl 4-phenylbutanol [4 (

5) One mouth] One L-proline t ert —Butyl amide (Example number 12) ^) ο

Next, a method for producing the compound of the present invention will be described.

CT / JP95 / 02546

(Method A)

In the above process scheme, and R2 have the same meanings as above, and R3 and R4 are the same and represent a arylmethyl group which may be substituted with lower alkyl, lower alkoxy or hydroxyl group, or Represents a hydrogen atom, the other represents an amino group-protecting group other than arylmethyl group, and R5 represents a normal amino-protecting group.

R ³ and R ⁴ are substituted with lower alkyl, lower alkoxy or hydroxyl group Examples of benzyl and arylmethyl groups include benzyl, 4-methylbenzyl, 4-methoxybenzyl, 4-ethylbenzyl, 4-ethoxybenzyl and 4-hydroxybenzyl. .

The “protecting group for amino groups other than arylmethyl group” for R 3 or R 4 and the usual protecting group for the amino group for R 5 are not limited as long as they are commonly used. Preferably, said "aliphatic acyl group"; benzoyl, o-naphthoyl.

^ Aryl carbonyl groups, such as 1-naphthyl, 2-bromobenzoyl, halogenylarylcarbonyl groups, such as 4-cyclobenzoyl, 2,4,6—trimethylbenzoyl, 4-toluoyl Lower alkylated aryls such as lower carbonyl groups, lower alkoxylated aryls such as 4-anisole, and lower alkoxylated aryls such as 4-anisole. Aromatic carbonyl groups, lower alkoxycarbonylated arylcarbonyl groups such as 2- (methoxycarbonyl) benzoyl, and arylated aryloxy groups such as 4-phenylbenzyl Acyl group; lower alkoxy such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, and isobutoxycarbonyl Lower alkoxycarbonyl substituted by halogen or tri-lower alkylsilyl groups such as cyclocarbonyl group, 2,2,21-triethoxy carbonyl, 2-trimethylsilylethoxycarbonyl An alkoxycarbonyl group such as a group; an alkenyloxycarbonyl group such as vinyloxycarbonyl and aryloxycarbonyl; benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, and 3,4-dimethoxybenzylo The aryl ring is substituted with one or two lower alkoxy or nitro groups, such as oxycarbonyl, 2-2 benzyloxycarbonyl, carbonyl, 412 benzyloxycarbonyl; Aralkyl carbonyl group; trimethylsilyl, triethylsilyl, isov Chirushi Li Le, t one-butyldimethylsilyl Shi Li Le, Mechirujii Sopuro pin Resid Li Le main Chiruji t - Buchirushi Li Le, cormorants good bet Li Lee Sopu port Pirushi Li Le Tri-lower alkyl substituted with one or two aryl groups, such as trialkyl lower alkylsilyl, diphenylmethylsilyl, diphenylbutyryl, diphenylsilyl, bilsilyl, and phenylsilyl propylsilyl It is a silyl group such as an alkylsilyl group.

Compound (VI) used as a starting material is known or can be synthesized according to a known method. The 4-chloroproline of compound (VI) can be obtained by using 4-hydroxyproline as a raw material, using hydrochloric acid, using phosphorus trichloride, or using an acid halide such as thionyl chloride or oxychlorine. Preferably, according to the method of AK Bose et al. [Tetrahedron Letters, 40, 3937 (1973), a method using N-chlorosuccinic acid imide and trialkyl phosphate, JG Calzada S (0rg. Syn. 634), a method of reacting carbon tetrachloride with triphenylphosphine is used.

Compound (V) used as a starting material is known or is easily synthesized according to a known method. For example, it is synthesized according to the method of R. Horanz et al. (Synthes is, 703, 1989) and the method of MT Reets et al. (Tetraheron Lett., 29.3295, 1988).

These are known compounds or compounds that can be easily synthesized according to known methods, and variations that are not detailed in this specification can also be used.

Hereinafter, each step will be described in detail.

The first step is a step of reacting the compound (V) or a reactive derivative of the carboxylic acid thereof with the amino compound (VI) to produce the compound (VII).

The reaction is carried out according to a general peptide synthesis method, for example, an azide method, an active esterification method, a mixed acid anhydride method or a condensation method.

In the azide method, amino acid hydrazine produced by reacting an amino acid or its ester with hydrazide in an inert solvent at around room temperature was reacted with a nitrite compound to convert it to an azide compound. Later, it is carried out by treating with an amine compound. Examples of the nitrite compound used include an alkali metal nitrite such as sodium nitrite or an alkyl nitrite compound such as isonitrite.

Examples of the inert solvent used include amides such as dimethylformamide and dimethyl acetate, sulfoxides such as dimethyl sulfoxide, and pyrrolidone such as N-methylpyrrolidone. Can be mentioned.

The two reactions in this step are usually performed in one reaction solution, and the reaction temperature is 150 to 0 in the first stage, 110 to 10 in the second stage, and the reaction time is Is 5 minutes to 1 hour, and the latter is 10 hours to 5 days. The active esterification method is carried out by reacting an amino acid with an active esterifying agent in a solvent to produce an active ester, and then reacting the active ester with an amide compound.

The solvent used is not particularly limited as long as it is inert. Examples thereof include halogenated hydrocarbons such as methylene chloride and chloroform, ethers, and tetrahydrofuran. Examples of such ethers include amides such as dimethylformamide and dimethylacetamide.

The active esterifying agents used include, for example, N-hydroxysuccinimide, n-hydroxybenzotriazole, N-hydroxy-5-norbornene-2,3-dicarboximide. N-hydroxy compounds can be mentioned, and the active esterification reaction is suitably carried out in the presence of a condensing agent such as dicyclohexylcarbodiimide (DCC). ,,,,, ,,,,,,,,,,,,,,,,,,,,,,,,,,,,-,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, ), N, N'-bis (2-oxo3'-oxazolidinyl) phosphonic chloride (B〇P-CI), NN'-carbonylimidazole, N, N'—Jisak Kiserate (DSO), N, N'-diphthalimidoxalate (DP〇), N, N'-bis (norbornenylsuccinimidyl) oxalate (BN0), 1,1'-bis (Benzotriazolyl) oxalate (BBTO), 1,1'-bis (6-benzobenzoylazolyl) oxalate (BCT 0), 1,1'-bis (6-trifluoromethylbenzoate) Condensation can also be carried out in the presence of a condensing agent of liazolyl) oxalate (B-B0) and bromitol tris-pyrrolidinosine phosphonium-hexafluoro-phosphate (PyBr OP).

The reaction temperature is between 10 and 25 c in the active esterification reaction, around room temperature in the reaction between the active ester compound and the amine, and the reaction time is between 30 min and 10 h for both reactions. is there.

The mixed acid anhydride method is carried out by producing a mixed acid anhydride of an amino acid and then reacting it with an amine.

The reaction to produce the mixed anhydride is carried out in an inert solvent (for example, ethers such as ether and tetrahydrofuran, amides such as dimethylformamide and dimethylacetamide), and ethyl ethyl carbonate. Lower alkyl carbonates such as sodium butyl carbonate, di-lower alkyl cyanophosphoric acid such as getyl cyanophosphoric acid (DEPC), or diphenyluric acid azide (DP PA) This is achieved by reacting the acid.

The reaction is suitably carried out in the presence of an organic amine such as triethylamine, N-methylmorpholine, the reaction temperature is between 110 and 25 "C, and the reaction time is 3 minutes. 0 minutes to 5 hours.

The reaction of the mixed anhydride with the amine is preferably carried out in an inert solvent (eg, ethers such as ether, tetrahydrofuran, amides such as dimethylformamide, dimethylacetamide). The reaction is carried out in the presence of the above-mentioned organic amine, the reaction temperature is from 0 to room temperature, and the reaction time is from 1 hour to 24 hours. The condensation method is carried out by directly reacting an amino acid and an amine in the presence of a condensing agent such as dicyclohexylcarbodiimide or carbonyldimidazole to produce the above-mentioned active ester. The reaction is performed in the same manner as in the reaction. The second step is a step of removing the protecting group for the amino group of the compound (VII) in a solvent to produce the compound (VIII).

When a silyl group is used as a protecting group for an amino group, it is usually treated with a compound that generates fluorine anion such as tetrabutylammonium fluoride or potassium fluoride. Removed.

The reaction solvent is not particularly limited as long as it does not inhibit the reaction, but ethers such as tetrahydrofuran and dioxane are preferable.

The reaction temperature and reaction time are not particularly limited, but the reaction is usually carried out at room temperature for 10 to 18 hours.

When an alkoxycarbonyl group such as t-butyloxycarbonyl group is used as a protecting group for an amino group, it is removed by treating with an acid in an inert solvent. .

The reaction solvent is not particularly limited as long as it does not inhibit the reaction. Amides such as dimethylformamide and dimethylacetamide; getyl ether, diisopropyl ether, and tetrahydrofuran Ethers such as dioxane, dimethoxetane, diethylene glycol dimethyl ether; methanol, ethanol, n-prono. Nor, Isoprono ,. Nol, n-butanol, isobutanol, t-butanol, isoamyl alcohol, polyethylene glycol, glycerin, octanol, cyclohexanol, methyl methyl ester Alcohols such as Solve, halogenated hydrocarbons such as dichloromethane, chloroform, and trichloroethane are preferred.

The acid to be used is not particularly limited as long as it is an acid, but is preferably an inorganic acid such as hydrochloric acid, an organic acid such as trifluoroacetic acid, or a trifluoroboronate ester. It is such a Lewis acid. The reaction temperature and reaction time are not particularly limited, but are usually 0 to 30 and the reaction is carried out by treating for 20 minutes to 1 hour.

When the protecting group for the amino group is an aliphatic acyl group, an aromatic acyl group, or a substituted methylene group forming a Schiff base, treatment with an acid or a saturated group in the presence of an aqueous solvent. Can be removed.

The acid used is not particularly limited as long as it is usually used as an acid. Preferably, an inorganic acid such as hydrochloric acid, sulfuric acid, phosphoric acid, or hydrobromic acid is used. The base used is not particularly limited as long as it does not affect the other parts of the compound, but is preferably a metal alkoxide such as sodium methoxide, or carbonate. Alkali metal carbonates, such as sodium and potassium carbonate, alkali metal hydroxides or ammonium hydroxide, such as sodium hydroxide and potassium hydroxide, ammonia and methanol Such ammonias are used.

It should be noted that isomerization may occur in hydrolysis with a base.

The solvent used is not particularly limited as long as it is one used in a usual hydrolysis reaction. Water; alcohols such as methanol, ethanol, n-propanol, tetrahydrofuran, and dioxane; Organic solvents such as ethers or mixed solvents of water and the above organic solvents are preferred.

The reaction temperature and reaction time vary depending on the starting material, the base used, and the like, and are not particularly limited. However, in order to suppress a side reaction, the reaction is usually performed at 0 to 150 for 1 to 10 hours.

When the protecting group for the amino group is an aralkyloxycarbonyl group, it is removed by reduction in an inert solvent in the presence of a catalytic reduction catalyst. The solvent to be used is not particularly limited as long as it is one used in a usual reduction reaction. Preferably, it is dimethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, dimethoxeta. Diethylene glycol

Ethers such as dimethyl ether; methanol, ethanol, n -Prono, 'Nol, Isoprono ヽ', n-butanol, isobutanol, toluene, isoamyl alcohol, diethyl glycol, glycoline, octanol , Alcohols such as cyclohexanol and methyl sorb.

Examples of the catalytic reduction catalyst to be used include palladium carbon, palladium black and the like.

The reaction temperature and reaction time vary depending on the starting materials and the like, and are not particularly limited. Usually, the reaction is carried out by reacting hydrogen at normal pressure to 10 atm near room temperature for 1 hour to 8 hours.

When the protecting group of the amino group is an arylmethyl group, it is usually removed by contacting with a reducing agent in a solvent, and preferably, the catalytic reduction is carried out at room temperature under a catalyst. It is preferable to use a removing method or a removing method using an oxidizing agent.

The solvent used in the removal by catalytic reduction is not particularly limited as long as it does not participate in the reaction. Alcohols such as ethanol, methanol, ethanol, and isopropanol; ethers such as getyl ether, tetrahydrofuran, and dioxane; and aromatics such as toluene, benzene, and xylene. Aliphatic hydrocarbons such as aromatic hydrocarbons, hexane and cyclohexane, esters such as ethyl acetate and propyl acetate, fatty acids such as formic acid and acetic acid, or a mixed solvent of these organic solvents and water Alternatively, a mixed solvent of a fatty acid and an alcohol is preferable.

The catalyst to be used is not particularly limited as long as it is usually used in a catalytic reduction reaction, but is preferably palladium black, palladium carbon, Raney-nickel, platinum oxide, platinum oxide Black, rhodium aluminum monoxide, triphenylphosphine rhodium monochloride, no. Radium monosulfate barrier is used. Although the pressure is not particularly limited, it is usually carried out at a pressure of 1 to 0 atm.

The reaction temperature and reaction time vary depending on the starting material and catalyst type, etc. It is usually carried out at 0 to 100 hours for 5 minutes to 24 hours.

The solvent used in the removal by oxidation is not particularly limited as long as it does not participate in this reaction, but is preferably a water-containing organic solvent.

Preferred examples of such organic solvents include ketones such as acetone, halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, and acetonitrile. Ethers such as nitriles, getyl ether, tetrahydrofuran, and dioxane; and amines such as dimethylformamide, dimethylacetamide, and hexamethylphosphorotriamide And sulphoxides such as dimethylsulphoxide.

The oxidizing agent to be used is not particularly limited as long as it is a compound usually used for oxidation, but preferably, potassium persulfate, sodium persulfate, or ammonium silicate is used. CAN) and 2,3-dichloro-5,6-dicyano- _p- benzoquinone (DDQ) are used.

The reaction temperature and reaction time vary depending on the starting material and the type of catalyst, etc., but are usually 0 to 150, and are usually from 10 minutes to 24 hours.

When the protecting group of the amino group is a diarylmethyl group, it can be achieved by performing the treatment in the same manner as in the above-mentioned conditions for the removal reaction of the arylmethyl group. When the amino-protecting group is an alkenyloxycarbonyl group, the amino-protecting group usually forms the above-mentioned aliphatic acyl group, aromatic acyl group, alkoxycarbonyl group or Schiff base. This can be achieved by treating with a base in the same manner as the conditions for the removal reaction in the case of a substituted methylene group. In addition, in the case of aryloxycarbonyl, the method of removing it using palladium and triphenylphosphine or nickel tetrapropyl is particularly simple and can be carried out with few side reactions.

The third step is a step of producing the compound (X) by reacting the compound (IX) or a reactive derivative of the carboxylic acid thereof with the compound (VIII) in the same manner as in the first step. The fourth step is a step of producing the compound (XI) by removing the protective period of the amino group of the compound (X) in the same manner as in the second step.

The fifth step is a step of reacting the compound (XII) or a reactive derivative of the carboxylic acid thereof with the compound (XI) in the same manner as in the first step to produce the compound (I) of the present invention.

(Method B)

In the above formula, R1, _R2 , _R3 , _R4 and R5 have the same meanings as described above, and R6 represents a hydroxyl-protecting group.

Hereinafter, each step will be described in detail. The sixth step is a step of producing a compound (XIV) by protecting the hydroxyl group of the compound (VII) by a usual method.

The seventh step is a step of producing a compound (XV) by removing the amino group of the compound (XIV) in the same manner as in the second step of the method A.

The eighth step is a step of reacting the compound (XVI) or a reactive derivative of the carboxylic acid thereof with the amino compound (XV) in the same manner as in the first step of Method A to produce the compound (XVII). is there.

The ninth step is a step of producing a compound (XVIII) by removing the protective period of the amino group of the compound (XVIII) in the same manner as in the second step of Method A.

In the tenth step, the compound (XII) or a reactive derivative of the carboxylic acid thereof and the amino compound (XVIII) are reacted in the same manner as in the first step of the method A to produce a conjugate (XIX). It is a process.

The first step is a step of reacting compound (X) with compound (XIX) to produce compound (XXI).

The 12th step is a step for producing the compound (I) of the present invention by selectively removing the hydroxyl-protecting group (R ⁶ ) of the compound (XXI).

After completion of the reaction in each of the above steps, each target compound is collected from the reaction mixture according to a conventional method. For example, the reaction mixture is appropriately neutralized, and if insolubles are present, they are removed by filtration, an organic solvent immiscible with water is added, and after washing with water, the solvent is distilled off. can get. The obtained target compound can be further purified, if necessary, by a conventional method, for example, recrystallization, reprecipitation or chromatography.

The administration form of the compound (I) of the present invention includes, for example, oral administration by tablets, capsules, granules, powders or syrups, or parenteral administration by injections or suppositories. These preparations may include excipients (eg, lactose, sucrose, dextrose, mannite, saccharides such as sorbite; corn starch, starch starch, starch, Dextrin, power Starch derivatives such as ropoxymethyl starch; cellulose derivatives such as crystalline cellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium, and internally-carboxylated methylcarboxymethylcellulose sodium. Arabic gum; dextran; organic excipients such as burlan: and silicate derivatives such as light silicic anhydride, synthetic aluminum silicate, and magnesium metasilicate magnesium; like calcium phosphate Inorganic excipients such as phosphates, carbonates such as calcium carbonate, and sulfates such as calcium sulfate. ), Lubricants (eg, stearic acid, calcium stearate, metal stearate such as magnesium stearate; talc; colloid silica; veegum; Cousins; boric acid; adipic acid; sulfates such as sodium sulfate; glycol; fumaric acid; sodium benzoate; DL leucine; sodium fatty acid salt; sodium lauryl sulfate; Lauryl sulfates such as magnesium lauryl sulfate; silicic acids such as silicic anhydride and silicic acid hydrate; and the above-mentioned starch derivatives.), Binders (for example, polyvinylpyrrolidone) Don, Macrogol, and the same compounds as the excipients can be mentioned.), Disintegrants (for example, the same compounds as the excipients, and Chemically modified starches such as croscarmellose sodium, carboxymethyl starch sodium, and cross-linked polyvinyl pyrrolidone can be mentioned.) Stabilizers (methylparaben, propylparaben) Paraoxybenzoic acid esters such as phenol; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonidum chloride; phenols such as phenol and cresol; Thimerosal; dehydroacetic acid; and sorbic acid), flavoring agents (for example, commonly used sweeteners, acidulants, flavors, etc.). It is manufactured by a well-known method using additives such as a diluent. The amount used depends on the symptoms, age, administration method, etc.For example, in the case of oral administration, the lower limit is 0.1 mg / kg and the upper limit is 20 mg / kg. In the case of intravenous administration, it is preferable to administer once or less, 0.01 mg / kg as the lower limit and 2 mg / kg as the upper limit, once or several times a day depending on the symptoms. The present invention will be described in more detail with reference to examples.

(Example)

(Example 1)

2 (S), 3 (S) -3- (N- (quinoxalin-2-carbonyl) -l-Lthreonyl 1_amino-1 2 --_ hydroxy-1 4-phenylbutanol-1 [4 (S) 1-mouth] 1 L-proline tert-butyl amide

1) 2 (S), 3 (S) — 3 — (N-tert-butoxycarboxy-L-leuronyl) _ _ _ MINOH _2 _ HYDROXY-1-4-phenylbutanol-[ 4 (S) —Cross mouth ”! L-proline tert—Butylamide

3 (S) 1 amino 2 (S) hydroxy 4 1 phenylbutanol

4 (S) monochloride] Proline tert-butylamide (200 mg, 0.52 mmol) was dissolved in N, N-dimethylformamide (hereinafter abbreviated as DMF) 6 m], and N-tert-Butoxycarbonyl-1-L-threonine (114 mg, 0.52 mmol) was added, and the mixture was cooled with water. 0.1 ml (0.624 millimol) of getyl cyanophosphate (hereinafter abbreviated as DEPC) was added, followed by dropwise addition of 0.14ral (1.04 millimol) of triethylamine, followed by stirring for 3 hours. The reaction mixture solvent was evaporated under reduced pressure, acetic Echiru 2 0 m 1 was added 5% Kuen acid, washed with 5 ⁰ / oN a HC O3 solution and saturated brine, and dried over anhydrous N a 2 S 0 ₄ . The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylennomethanol chloride = 8: 1). 90 mg of the title compound as a colorless crystalline solid was obtained.

Melting point 2 1 1-2 1 4 "Co

Elemental analysis: C ₂₈ H ₄ 3 ^ 0 ₇ C1 · 1 / 2H ₂₀ (molecular weight 592.12) Theory: C, 56.79; H, 7.49; N.9.46; C1, 5.99

Found: 56.93; H ₍ 7.17;, 9.46; C1, 6.07.

Mass s: 584 (M + H) +.

NMR: δ (dg-D SO, 270 MHz): 7.97 (1H, d, J = 7.9 Hz), 7.64 (1H, s), 7.41 (2H, d, J-6.6 Hz), 7.28-7.18 (3H , m), 6.36 (1H, d, J = 8.6Hz), 5.01 (1

H, d, J = 7.3Hz), 4.71 (lH, d, J = 5.3Hz), 4.55 (lH, m), 4.43 (1H, t, J = 7.9Hz), 4.34-4.19 (3H, m), 3.81-3.61 (3H, m), 2.82-2.65 (3H, m), 2.01-1.96 (lH, m), 1.43 (9H, s), 1.30 (9H, s), 0.86 (3H, d, J = 5.9Hz).

2) 2 (S), 3 (S) 1-3-[N-(quinoxaline 1-2-carbonyl)-1-L-threonyl] amino 2-hexane-1-phenyl-1-4-phenylbutanol (S) 1-mouth] 1-L-proline tert-butylamide

2 (S), 3 (S) 1 3— (N-tert-butyloxycarbonyl L-threonyl) amino 2-hydroxy 4-phenylbutanol [4 (S) —close-mouth] 1 0.33 g (0.57 mmol) of L-proline tert-butylamide was added to 10 ml of a 4 N hydrochloric acid-dioxane solution, and tert-butyloxyl-ponylol was removed. The total amount of the obtained hydrochloride and 2-quinoxali Carboxylic acid (99 mg, 0.57 mmol) was dissolved in DMF (6 ml) and cooled with ice. DEPCO.11m] (0.684 millimoles), followed by triethylamine 0.16m1 (

I.14 mimol) was added dropwise and stirred for 3 hours. The reaction mixture was removed distillate solvent under reduced pressure, acetic Echiru 2 0 m 1 was added 5% Kuen acid, washed with 5% N a HC 03 aqueous solution and saturated brine, and dried over anhydrous N a ₂ S 0 _4. The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride = 8: 1). Crystallization from hexane monoethyl ether gave 244 mg of the title compound as a white crystalline solid.

Mp 109-118.

Elemental analysis: as the _{_{_{_{C 32 H 39 N 6 0 6}}}} CI ( molecular weight 639. I ⁴⁾

Theoretical value: C, 60.13; H, 6.15: N, 13.15; C1.5.55 Found: C, 59.91; H, 6.09; N, 13.04; C1, 5.64.

Mass: 639 (M + H) ⁺ .

NMR: δ (CD3OD, 270MHz): 9.50 (lH, m), 8.23-8.16 (2H, m), 7.99-793 (2H, m), 7.33-7.29 (2H, m), 7.13-7.14 (2H, ra), 7.04-7.01 (1H, m), 4.47-4.34 (6H, m), 4.23-4.19 (1H, m), 3.78 (1H, m), 3.01 (1H, dd, J = 13.9, 3.3 Hz), 2.79-2.70 (2H, m), 2.11-2.06 (lH, m),] .36, 1.30 (9H, ss),

1.17 (3H, d, J = 6.6Hz).

(Example 2)

2 (S), 3 (S) -3- (N- (quinoxaline-l-carbonyl) -l-O-acetyl-L-threonyl] _amino-2-hydroxyl-l-phenylbutanoyl [4 (S) one mouth] one; L-proline tert-butylamine_

1) 2 (S), 3 (S) -3-([-benzyloxycarbonyl) -l-threonyl 1_amino-2- tert-dibutyldimethylsilyloxylinini 4-phenylbutanol [4 ( S) Monochrome] — L-proline tert — Butylamide

0.64 g (l.01 mimol) of the compound synthesized in Reference Example 2 was dissolved in 50 ml of methanol and hydrogenated with 5% palladium-carbon catalyst to remove the benzyloxycarbonyl group. Dissolve 0.53 g (1.52 mmol) of the obtained amino compound and N-benzyloxycarbonyl-L-threonyl-N-hydroxysuccinimide ester in 12 ml of DMF, cool with water, and add triethylamine 0.2 Sral (2.02 mmol) was added dropwise and stirred for 5 hours. The reaction mixture was evaporated reduced pressure solvent, acetic Echiru 3 0 m 1 was added 5% Kuen acid, 5% N a HC_〇 ₃ solution and saturated brine in washing诤, and dried over anhydrous N a ₂ S 0 ₄ . The solvent was distilled off under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride Z methanol =] 0:]). Crystallization with hexane gave 0.46 g of the title compound as a white crystalline solid. NMR: δ (CDC13, 270MHz): 7.36-7.13 (10H, m), 6.49-6.42 (1H, m), 5.47 (lH.d, J = 8.3Hz). 5.13 (2H, q, J = 25.2, 12.0) Hz), 441-416 (6H, m), 3.96 (1H, dd, J = 8.5, 1.7Hz), 3.75 (lH, m), 3.20 (1H, dd, J = 14.0, 3.5Hz) , 3.00 (1H, d, J = 3.7Hz), 2.68-2.43 (3H, m), 1.31 (9H, s), 1.10 (3H, d, J-6.5Hz), 0.96 (9H, s), 0.13 ( (6H, d, J = 5.4Hz).

2) 2 (S), 3 (S)-3-[N-(quinoxaline-1-carbonyl)-1-L-threonyl] amino 2-tert-butyldimethylsilyloxy 4-phenylbutanol [4 (S) 1-_1: purine tert-butylamide

2 (S), 3 (S) -13-[(N-benzyloxycarbonyl) -1-L-threonyl] amino 2-tert-butyldimethylsilyloxy-14-phenylbutanol [4 ( S) One-sided mouth] 0.37 g (0.50 mmol) of proline tert-butylamide is dissolved in 15 51 of methanol and hydrogenated with 5% palladium-carbon catalyst to form benzyloxycarbonyl. The radical was removed. The total amount of the obtained amino compound and 96 mg (0.55 mmol) of 2-quinoxalinecarboxylic acid were dissolved in 8 ml of DMF and cooled with ice. 098 ml (0.6 mmol) of DEP CO was added, and then 0.14 ml (1.00 mmol) of triethylamine was added dropwise, followed by stirring for 3 hours. The reaction mixture solvent was evaporated under reduced pressure, acetic Echiru 2 0 m 1 was added 5% Kuen acid, 5% N a HC_rei_3 solution and saturated brine in washing净, and dried over anhydrous N a 2 S 0 ₄ . The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride methanol = 7: 1) to obtain the title compound (306 mg).

NMR: S (CDCI3, 270MHz): 9.57 (1H, s), S.46 (1Η.d, J = 8.5Hz). 8.2 5-8.12 (2H, m), 7.94-7.87 (2H, m), 7.08 (2H, d, J = 7.5Hz), 6.S8 (2H, m).

6.77-6.59 (3H.m), 4.51-4.27 (7H.m) .3.81-3.78 (1H, m), 3.63 (1H, br.s), 3.24-3.17 (1H, m) .2.75-2.67 (1H m), 2.60-2.51 (2H, m) 1.32 (9H.s), 1.19 (3H, d, J = 6.5Hz) .0.96 (9H, s), 0.15 (6H, d.J = 3.9Hz) . 3) 2 (S), 3 (S) 1-3-[N-(quinoxalin-1-carbonyl) 1-acetyl-L-threonyl] ammino-1-2-tert-butyl-dimethyl 4 1-phenylbutanol [4 (S) chloro] 1-L-proline tert-butylamide

2 (S), 3 (S) -13- [N- (quinoxaline-12-carbonyl) -1L-threonyl] amino-2-tert-butyldimethylsilyloxy-14-4phenylbutyryl [4 (S) one mouth] — 100 mg (0.13 mmol) of L-proline tert-butylamide was dissolved in 4 ml of methylene chloride, 0.025 ml (0.26 mmol) of anhydrous acetic acid was added, and then 4-dimethylamino 1 mg (0.013 mmol) of pyridin was added and the mixture was stirred at room temperature for 1 晚. The reaction mixture solvent was evaporated under reduced pressure, acetic acid Echiru 2 0 m I added 5% Kuen acid, 5 6 N a HC 0 ₃ washed with an aqueous solution and saturated brine, and dried over anhydrous N a ₂ S 0 _4. The solvent was filtered under reduced pressure to obtain the title compound (65 mg).

M a s s: 795 (+ H) +

NMR: δ (CDCI 3.270MHz): 9.61 (1Η, s), 8.36 (1H, d, J = 9.2Hz), 8.2 4-8.16 (2H, ra) 7.95-7.90 (2H, m), 7.26 -7.15 (2H, m), 7.12-7.01 (2H, m).

6.85-6.73 (2H, m), 6.61 (lH, br.s), 5.47-5.42 (1H, m), 4.65-4.59 (1H, m), 4.47-4.2δ (5Η, m), 3.71-3.70 ( 3.11-3.05 (lH, m), 2.71-2.58 (3H, m), 1.96 (3H, s), 1.31 (9H.s), 1.26 (3H, d, J = 5.9Hz) , 0.92 (9H, s), 0.10 (6H, d, J = 8.6Hz).

4) 2 (S), 3 (S)-3-[N-(quinoxaline-1-carbonyl)-0-acetyl-L-threonyl 1-amino 2-hydroxy 4-phenylbutanol One [4 (S) one mouth] One L-proline tert-butyl amide

2 (S), 3 (S) — 3 — [N- (quinoxaline- 1 2 — carbonyl) -1 0-acetyl-L-threonyl] amino-2- 4-tert-butyldimethylsilyloxy 1 4-phenylbutanoy Lou [4 (S) 1 black mouth] 1 Loop / 94

twenty five

Dissolve 65 mg (81.7 micromol) of tert-butyl amide in mouth in 12 ml of tetrahydrofuran (hereinafter abbreviated as THF), and add 28 mg (0.49 mimol) of tetral n-butylammonium fluoride I ) Was added and the mixture was stirred at room temperature for 14 hours. Acetate Echiru 2 0 ml was added, washed诤with 5% N a HC 0 ₃ solution and dried over anhydrous N a 2 S 0 _4. The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride / methanol = 9: 1). Crystallization from hexane monoethyl ether gave 23 mg of the title compound as a white crystalline solid.

With a melting point of 10 8—1 12

Elemental analysis: C34H4iN ₆ 0 ₇ as a Cl · H ₂ 0 (molecular weight 6S1.17)

Theory II: C, 59.95; H, 6.07; N, 12.34; CI, 5.20

Found: C, 60.28; H, 6.23; N, 12.05: CI, 5.01,

a s s: 681 (M + H) +.

NMR: δ (CDCI3, 270 MHz): 9.63 (1H, s), 8.60 (1H, d, J = 8.5 Hz), 8.2 5-8.19 (2H, m), 7.94-7.86 (2H, m), 7.28-7.11 (5H, m), 7.02-6.96 (1H, m) .6.44dH.br), 5.55-5.51 (1H, ra), 4.88-4.83 (1H, m), 4.58-4.52 (2 H, m), 4.36 -4.23 (2H, m), 4.04-3.99 (1H, m), 3.84-3.77 (lH, m), 2.88 -2.53 (4H, m), 2.05, 2.04 (3H, ss), 1.85-1.79 (1H, m) .1.32 (3H, d, J = 6.4Hz), 1, 37, 1, 28 (9H, ss).

(Example 3)

2 (S), 3 (S) -1-3- [N- (quinoxaline-12-carbonyl) -10-methyl-L-threonyl] amino-2-hydroxy-4-phenylbutanoyl [4 ( S) Monochrome] — L-proline tert-butyl 7-medium 2) (S), 3 (S) -3-(N-tert-butyloxycarbonyl) 0-notyl-L-leonich Amy _ 2 _— ^ key J "4-Feline butanol 4_ (S) — black mouth] — L-proline tert — butyramite 3 (S) -Amino 2 (S) -Hydroxy 4 4-Phenylbutanoyl [4 (S) -Cro-mouth] Proline tert-Butylamide 190 mg (0.50 mmol) in methylene chloride 6m 1, and added to the N-tert-butoxycarbonyl group synthesized according to the method of Francis MF Chen and Leo Benoiton (J. Org. Chem., 1979, vol. 44, 2299). Methyl-L-threonine (114 mg, 0.52 mmol) was added and cooled on ice. 98 mg (0.56 mimol) of DEPC was added, followed by dropping of 57 mg (0.56 mimol) of triethylamine, followed by stirring for 3 hours. The reaction mixture solvent was evaporated under reduced pressure, acetic Echiru 2 0 m 1 was added 5% Kuen acid, washed with 5% N a HC_rei_3 solution and saturated brine, and dried over anhydrous N a ₂ S 0 _4. The solvent was concentrated under reduced pressure, and ethyl ether was added to the oily residue, and the resulting crystals were collected by filtration. Recrystallization from ethyl ether gave 280 mg of the title compound as colorless fine needles.

Melting point 2 08—2 1 1.

Elemental analysis: as a C29H45N4O7CI · H ₂ 0 (molecular weight 615.17)

Theoretical value: C, 56.62; H, 7.70; N, 9.11; C1, 5.76

Found: C, 56.68; H, 7.11; N, 9.31; C1, 6.51.

M a s s. * 596M +.

NMR: o (CD3OD, 360MHz): 7.33-7.14 (5H, m), 4.44-4.31 (4H, m), 3.98-3.94 (1H, m), 3.76 (1H, dd, J = 10.2, 8.0Hz) , 3.66-3.60 (1H.m), 3.12 (3H, s), 2.92-2.70 (3H, m), 2.17-2.10 (1H, m), 1.45 (9H.s), 1.32 (9H, s), 1.08 -1.03 (1H + 3H, d (3H), J = 6.3Hz).

2) 2 (S), 3 (S)-3-[N-(quinoxalin-1-carbonyl)-0-methyl-1-L-s-leonyl] amino 2-hydroxy-1-4-butane Nooru [4 (S) black mouth] — L-proline tert—butylamite

2 (S), 3 (S) — 3 — (N— te 1-t — Butoxyloxycarbonyl 0 — Methyl-l-Lethreonyl) Amino 2 — Hydroxy-1 4 1-phenyl Tanoyl [4 (S) 1-mouth] 1-L-proline tert-butylamide 0.24 g (0.4 mimol) was added to 10 ml of 4 N hydrochloric acid-dioxane solution, and tert-butyrate was added. After removing the carbonyl, the total amount of the obtained hydrochloride and 77 mg (0.44 mimol) of 2-quinoxalinecarboxylic acid were dissolved in 6 ml of DMF and cooled with ice. 77 mg (0.44 mimol) of DEP C was added, and then 85 mg (0.84 mimol) of triethylamine was added dropwise, followed by stirring for 3 hours. The reaction mixture was evaporated under reduced pressure Solvent, acetate Echiru 2 0 m 1 was added 5% Kuen acid, 5% N a HC 0 one solution and saturated brine in washing诤, dried over anhydrous N a ₂ S 0 ₄ did. The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride = 10: 1). Crystallization from hexane and ethyl ether gave 194 mg of the title compound as a white crystalline solid.

90-92.

Elemental analysis: as the _{_{C 33 H4iN 6 06Cl '1/}} 2 Η 2 0 ( molecular weight ^{2 .1Θ)}

Theoretical: 59.86; Η, 6.39; Ν, 12.69; C1, 5.35

Found: 59.97; H, 6.57; N, 12.20; C1, 5.03.

M a s s: 653 (M + H) +.

NMR: δ (CD3OD, 360MHz): 9.53 (1H, m), 8.27-8.24 (lH, m) .8.23 -8.18 (1H, m), 8.01-7.95 (2H.m), 7.31

(1H, d, J = 7.7Hz), 7.24-7.20 (2H, m), 7.11 -7.07 (1H, m), 4.51 (1H, d, J = 3.5Hz), 4.47-4.34 (4H, m), 3.91-3.85 (1H, m), 3.77 (1H, dd, J = 10.4, 3.8Hz), 3.26 (3H, s), 2.99-2.93 (2H.m), 2.79-2.70 (2H.m), 2.15- 2.08 (1H, m), 1.28 (9H, s),) .14 (3H, d, J = 6.1Hz), 0.9

(1H, t, J = 6.7Hz).

(Example 4)

2 (S), 3 (S) -3-[[N- (quinoxaline-1-2-carbonyl) -1 0-methoxymethyl-1L-threonyl] amino 2-hydroxy-1 4-phenylbutanoy L- [4 (S) -black mouth] L-proline tert-butyl Rua Mid

1) 2 (S), 3 (S) 1 3-(N-benzyloxycarbonyl) 1 0

Nilbutanol (4 (S) 1-mouth) 1 L-proline t er t —Butyl amide

3 (S) -Amino 2 (S) -Hydroxy-1-4-Phenylbutanoyl [4 (S) -Cross-mouth] Proline tert-Butylamide 190 mg (0.50 mmol) The residue was dissolved in 6 ml of methylene chloride, and 149 mg (0.50 mmol) of N-benzyloxycarbonyl-10-methoxymethyl-L-threonine was added thereto, followed by cooling on ice. 96 mg (0.55 mimol) of DEP C was added, and then 56 mg (0.55 mimol) of triethylamine was added dropwise, followed by stirring for 3 hours. The reaction mixture was evaporated reduced pressure solvent, acetic Echiru 2 0 m 1 was added 5% Kuen acid, washed with 5% N a HC 0 ₃ solution and saturated brine, and dried over anhydrous N a ₂ S 0 _4. The solvent was reduced under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride / methanol = 20: 1). Crystallization from hexane monoethyl ether gave 260 rag of the title compound as a white crystalline solid. Mp 57.5-5-9.

Elemental analysis: as C ₃₃ H45N408C1 · H ₂₀ (molecular weight 679.2 ² )

Theory: 58.36; H, 6.97; N.8.25; C1.5.22

Obtained: C.58.33; H, 7.02: N, 8.21: C1, 5.11.

Mass: 661 (M + H) +.

NR: δ (CDCI3, 270MHz): 7.37- 7.01 (10H, m), 6.3 (1H, br.s), 5.64-5.59 (1H.m), 4.49 (3H, br.s), 4.45-4.07 (6H , m). 3.86-3.64 (2H, m), 3.24 (3H, s), 2.81-2.53 (5H, m), 1.3K9H, s), 1.1δ (2Η, d, J = 5.7Hz), 0.89 ( 1H.t, J = 5.1, 6.9Hz).

2) 2 (S), 3 (S)-3-[N-(quinoxaline-1-carbonyl) 1-0-methoxymethyl-L-_ threonyl-amino _-2-_ Hydroxy J_ One phenylbutanol [4 (S) one mouth "! _ One I One proline ter _t — Butylamide

2 (S), 3 (S) -13- (N-benzyloxycarbonyl) 0-methoxymethyl-L-threonyl synthesized in Example 4 Amino 2-hydroxyl 4-phenylbutanol [4 (S) 1-closed mouth] Dissolve 210 mg (0.3 mmol) of 1 L-proline tert-butylamide in 10 ml of methanol and hydrogenate with 10% palladium-carbon catalyst. The benzyloxycarbonyl group was removed. The total amount of the obtained amino compounds and 55 mg (0.33 mimol) of 2-quinoxalinecarboxylic acid were dissolved in 5 ml of DMF and cooled with ice. 58 mg (0.33 mimol) of DEP C was added, and then 64 mg (0.63 mimol) of triethylamine was added dropwise, and the mixture was stirred for 0 hours. Acetate Echiru 2 0 m I added 5% Beautique phosphate, washed with 5% N a HC 0 ₃ solution and saturated brine, and dried over anhydrous N a 2 S 0 4. The solvent was distilled off under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene nomanol = 20: 1). Crystallization from ethyl ether-hexane gave 60 mg of the title compound i as a white crystalline solid.

Melting point 8 2 — 8 8.

Elemental analysis: C ₃₄ H43N607C1 · 1 / 2H ₂₀ (molecular weight 692, 22)

Theoretical: C, 59.00; H, 6.41; N, 12.14; C1, 5.12

Found: 58.97; H, 7.02: N, 11.66: CI.5.05.

M a s s: 683 (+ H) +.

NMR: δ (CDC13, 360 MHz): 9.62 (1H.m), 8.7 (1H, d, J = 7.2 Hz), 8.23-8.20 (2H, m), 7.94-7.85 (2H, m), 7.31-7.11 ( 5H, m), 6.23 (1H.s). 4.74-4.66 (2H, m), 4.57 (1H, d, J = 2.4Hz), 4.50-4.2 (6H, m).

3.82 (1H.dd, J = 10.7, 6.8Hz), 3.69-3.56 (1H, m), 3.38 (1H, q,).

2.84-2.60 (5H, m). 1.39 (1H, s), 1.27 (9H, s), 1.04 (1H, d, J = 6. Hz), 0.9 (1H, t, J = 6.6, 6.1 Hz). (Example 5)

2 (S), 3 (S) -3-3- [N— (quinoxaline-12-carbonyl) -10-succinyl-L-threonyl] amino 2-hydroxy-4 [4- (4-butaylyl)] S) Monochrome] — L-Proline tert-butyl amide

1) 2 (S), 3 (S)-3-[(N-benzyloxycarbonyl) 1-L-threoni ^ amino 2-methoxymethyloxy 4-phenylbutanol 4 ( S) 1-mouth] 1-L-proline tert-butyl amide

1.90 g (3.40 mimol) of the compound synthesized in Reference Example 3 was dissolved in 50 ml of methanol, and hydrogenated with 10% palladium-carbon catalyst to remove a benzyloxycarbonyl group. The total amount of the obtained amino compound and N-benzyloxycarbonyl; L-threonyl-N-hydroxysuccinimide ester 1.24 g (3.70 mimol) were dissolved in methylene chloride (60 ml), cooled with ice, and cooled with ice. 0.56 ml (4.04 mmol) of tilamine was added dropwise and stirred for 16 hours. Anti応混compound solvent was evaporated under reduced pressure, acetic Echiru 3 0 m 1 was added 5% Kuen acid, 5% N a HC 03 solution and saturated brine in washing诤, dried over anhydrous N a 2 S 0 ₄ did. The solvent was evaporated under reduced pressure, and the oily residue was purified by silica gel chromatography (methylene chloride-methanol-methanol-10 :). Crystallization from hexane gave 1.56 g of the title compound as a white crystalline solid.

Melting point 8 3 — 8 6.

Elemental analysis: as C ₃₃ H45N408C1 (molecular weight 661.20)

Theory: 59.95; H, 6.86; N. 8.47; C1.5.36

Found: 59.75: H, 6.89; N, 8.46: C1, 5.44.

Mass: 661 (M + H) +.

NMR: δ (CDCI3, 270MHz): 7.38-7.24 (10H, m), 6.95 (1H, d, J = 4.8Hz), 6.26 (1H, br.s). 5.69 (1H, d, J = 4.5Hz) ), 5.ll (2H.d, J = 6.1Hz), 4.79-4.63 (3H.m), .52-4.06 (7H, m), 3.77-3.71 (1H.m). H, s), 2.95 (2H, d, J = 7.17Hz), 2.56 (2H, t, J = 7.1Hz), 1.31 (9H, s) 1.12 (3H, d, J = 6.5Hz).

2) 2 (S), 3 (S) 13-[N-(quinoxaline-12-carbonyl)-l-threonyl] amino 2-methoxymethyl-1-phenylbutanoyl [4 (S) 1-outlet ] 1 L 1 proline tert-butylamine on

2 (S), 3 (S) 1 3 — [(N-benzyloxycarbonyl) 1 L-threonyl] amino 2- 2-methoxymethyloxy 4-phenylbutanoyl

[4 (S) 1-mouth] Dissolve 1.55 g (2.35 mimol) of proline tert-butyl amide in 40 ml of methanol and hydrogenate with 10% palladium-carbon catalyst to benzyloxycarbonyl group. Was removed. The total amount of the obtained amino compounds and 412 mg (2.37 mmol) of 2-quinoxalinecarboxylic acid were dissolved in 40 ml of DMF and cooled with ice. 1-Ethyl-3 (3-dimethylaminobu pill) carbodimid hydrochloride 450 mg (2.35 mimol), then 1-hydroxybenzotriazo azo-luo hydrate 317 mg (2.35 mimol) were added, and the mixture was stirred for 14 hours. did. The reaction mixture solvent was evaporated under reduced pressure, acetic Echiru 2 0 ml was added, 5% Kuen acid, 5% N a HC 03 solution and saturated brine in washing诤and Drying over anhydrous N a ₂ S 0 ₄ . The solvent is decompressed and the oily residue is purified by silica gel chromatography.

(Methylene chloride Z methanol = 10: 1) to give the title compound (1.12 g).

Melting point 8 2 — 8 7;

Elemental analysis: as the C ₃₄ H43N607C Bok H ₂ 0 (molecular weight § ^01.23)

Theoretical value: C, 58.24; H, 6.47; N, 11.98; C1, 5.06

Found: 57.87: H.6.73; N.11.87: CI, 4.62.

Mass: 683 (M + H) ⁺ .

NMR: δ (CDC13, 270MHz): 9.65-9.55 (1H, m), 8.31-7.83 (6H, m) .7.37-7.25 (5H, m), 7.01 (1H, d, J = 7.4Hz), 6.35 (1H, br.s), 4.79 -4.21 (9H, ra), 3.79-3.73 (1H, m). 3.37 (3H, s), 3.08-2.83 (2H, m), 2.72-2.51 (2H, m), 1.31 (9H, s). 1.21 (3H, d, J = 6.5Hz).

3) 2 (S), 3 (S) -3- (N- (quinoxaline-l-2-carbonyl) -1 0-succinyl-L-threonyl] amino 2- 2-methoxymethyloxy 4-phenyl Evening Root [4 (S) 1-mouth] 1 L-proline tert-butylamide 2 (S), 3 (S) 13-[N-(quinoxaline-1-carbonyl) 1 L —Threonyl] amino 2 —Methoxymethoxyloxy 4-phenylphenyl [4 (S) monochloride] — L-proline tert-butylamide 1, llg (l.63 mimol) Was dissolved in methylene chloride (20 ml), 360 mg (3.60 mmol) of succinic anhydride was added, then 3 mg (0.039 mmol) of 4-dimethylaminopyridine was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture solvent was evaporated under reduced pressure, acetic Echiru 2 0 m 1 was added 596 Kuen acid, 5 6 N a HC O3 solution and a saturated saline wash诤, and dried over anhydrous N a 2 S 0 _4. The solvent was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (methylene chloride = 10: 1) to obtain 756 mg of the title compound.

Melting point 99-101.

Elemental analysis: As C ₃₈ H ₄ 7N60ioCl (molecular weight 783.29)

Theory: 58.27: H, 6.05; N, 10.73; C 1.4.53

Found: 57.81; H, 6.28; N, 10.60; C1, 4.50.

Mass: 783 (M + H) ⁺ .

NMR: δ (CDCI3, 270 MHz): 9.61 (1H, s), 8.61 (1H ₍ d, J = 7.9 Hz).

8.23-8.19 (2H.m), 7.89-7.86 (2H, m), 7.26-7.22 (5H, m), 6.45-6.37 (1H, br.s), 5.41-5.38 (1H, m), 4.78 -4.58 (3H, m), 4.45-4.21 (4H, m), 3.71- 3.65 (1H, m), 3.47 (3H, s) .3.42 (1H, d, J = 8.6Hz), 3.36 ( 2H, s), 2.94-2.88 (2H, m), 2.66-2.50 (6H, m), 1.36- 1.27 (12H, m).

4) 2 (S), 3 (S) _- 3 -_ [N-(quinoxaline-1 2-carbonyl Amino 2-hydroxy 4-phenylbutanol [4 (S) 1-mouth] L-proline tert-butyramide

2 (S), 3 (S) 13- [N- (quinoxaline-12-carbonyl) monosuccinyl L-threonyl] amino 2- methoxymethyloxy 4-phenylbutano Yield [4 (S) 1-mouth] 1-L-Brolin tert-butylamide 270 mg (0.35 mmol) was dissolved in methylene chloride (20 ml), and trifluoracetic acid 0.2 ml (1.8 mmol) was added. And stirred at room temperature for 2 hours. Acetic acid Echiru 2 O ml was added, washed with 5% N a HC_rei_3 solution and dried over anhydrous N a 2 S 0 _4. The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride: ethanol: acetic acid = 95: 5: 3) to give 130 mg of the title compound as a white crystalline solid. Was.

With a melting point of 1 2 6 1 2 9.

Elemental analysis: (: ₃ 43 09 ^ '1.50 (molecular weight 766, 26)

Theoretical: C, 56.43; H, 6.05; N, 10.97; C 1,4.63

Found: C, 56.23; H, 6.06: N.10.23; C1, 4.36.

M a s s! 739 (M + H) +.

NMR: δ (CDC13-270MHz): 9.62 (1Η, s), 8.70-δ.59 (1H, m), 8.2 3-8.18 (2H, m), 7.93-7.84 (2H, m), 7.26-7.20 ( 5H, m), 7.02-6.97 (1H, m), 6.30 (1H, br.s), 5.54-5.50 (1H, m), 4.83-4.11 (7H, m), 3.97-3.68 (2H, m ), 2.97-2.51 (8H, m), 1.43- 1.22 (12H, m).

(Example 6)

2 (S), 3 (S) -1-3- [N- (quinoxaline-12-carbonyl) -1-L-aros-leonyl 1-amino-^^-hydroxy-4-phenylbutanoy "-[4 (S) 1-mouth] — L-proline tert-butylamide

1) _2_ (S) -3 (S) 13- (N-tert-butyloxycarbonyl-L-aros-leonyl) amino 2-hydroxy-14-phenylbutano Yl— “4 (S) -closed mouth” —L-proline tert-butylamide

3 (S) 1 amino 2 (S) 1-hydroxyl 4-phenylbutanol [

4 (S) 1 mouth] Proline tert-butylamide 461 mg (1.21 mimol) was dissolved in N, N-dimethylformamide (DMF) 10 ml 285 mg (1.30 mimol) of N-tert-butyloxycarbonyl-L-aros-leonin were added, and the mixture was cooled with ice. 0.23 ml (1.31 mimol) of getyl cyanophosphate (hereinafter abbreviated as DEPC) was added, and then 0.18 ml (1.31 mimol) of triethylamine was added dropwise, followed by stirring for 3 hours. The reaction mixture was evaporated reduced pressure solvent, acetic Echiru 2 0 m 1 was added 5% Kuen acid, 5% N a HC O3 solution and a saturated saline wash诤, and dried over anhydrous N a 2 S 0 _4. The solvent was concentrated under reduced pressure, and the oily residue was purified by silica gel chromatography (methylene chloride methanol = 10: 1). 670 mg of the title compound were obtained as a colorless crystalline solid.

Mp 194-1-196.

a s s: 583 (M) +.

NMR: δ (CDCl3, 270MHz): 7.32-7.12 (5H, m), 4.43-4.04 (9H, m), 3.96-3.73 (3H, m), 2.95-2.67 (4H, m), 2.18-2.07 (1H , m), 1.44 (9H.m), 1.33 (9H, s) 1.01 (3H, d, J = 6.1Hz)

2) 2 (S), 3 (S) 1-3-[N-(quinoxaline-1-carbonyl)-1-L-alos-leonyl] amino 2-Hydroxy-4-phenylbutanol-4 [S ) —H H_ mouth] — L-proline tert —Butylamide

2 (S), 3 (S) -13- (N-tert-butyloxycarbonyl-1L-arosthreonyl) amino-2-hydroxy-14-phenylbutanoyl [4 (S) 1-chloro Mouth] 1 L-Proline tert-butylamide 651 mg (l.12 mmol) was treated with 4 N hydrochloric acid-dioxane solution (10 ml) to remove tert-butyloxycarbonyl and obtained. The total amount of the hydrochloride and 193 mg (1.11 mmol) of 2-quinoxalinecarboxylic acid were dissolved in 0 m1 of DMF and cooled on ice. Was. 18 ml (1.18 mimol) of DEP CO was added, and then 0.42 ml (0.30 mimol) of triethylamine was added dropwise, followed by stirring for 16 hours. The reaction mixture solvent was evaporated under reduced pressure, acetic Echiru 0 0 m 1 was added 5% Kuen acid, 5% N a HC 03 solution and saturated brine in washing诤, and dried over anhydrous N a 2 S 0 _4. The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride Z methanol: = 10: 1). Crystallization from hexane ethyl ether gave 305 mg of the title compound as a white crystalline solid.

Melting point 1 1 7—1 2 1.

Elemental analysis: C32H39N6O6CI · 1.5 · ₂₀ (molecular weight 666.18)

Theoretical: 57.70; Η, 6.35; Ν, 12.62; C1, 5.32

Found: 57.97; H, 6.11; N, 12.31; C1, 5.01.

Mass: 639 (M + H) ⁺ .

NMR: δ (CDC 13.270 MHz): 8.50 (lH, d, J = 8.5 Hz), 8.24-8.12 (2H, m), 7.95-7.85 (2H ₍ m), 7.14-7.09 (2H, m), 6.99- 6.87 (2H, m), 6.77-6.71 (lH.m), 6.20 (1H, br.s), 454-4.42 (4H, m), 4.38-4.22 (3H, m), 4.11 -3.67 (5H , M>, 2.85-2.62 (4H, m), 1.30-1.21 (12H, m)

(Example 7) 2 (S), 3 (S) -3- (N- (quinolin-12-carbonyl) -1-L-threonyl 1_amino_2_2__h Nylor “4 (S) 1-channel] — L-proline tert — butylamide

2 (S), 3 (S) -1 3- (L-threonyl) amino 2-hydroxy-1 41-Phenylbutanoyl [4 (S) -closure] 1-L-proline tert- Dissolve 118 mg (0.25 mmol) of butylamide and 55 mg (0.31 mmol) of quinaldic acid in 5 ml of THF, cool with water, and cool with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride 57 mg. (0.3 mmol) and then 38 mg (0.28 mmol) of 11-hydroxybenzotriazole-hydrate were added, and the mixture was stirred for 6 hours. The solvent was distilled off from the reaction mixture under reduced pressure, 20 ml of methylene chloride was added, and the mixture was washed with 10% citric acid, 5% NaHC3 aqueous solution and saturated saline. 净, and dried over anhydrous N a 2 S 0 _4. The solvent was concentrated under reduced pressure, and the oily residue was purified by preparative thin-layer chromatography (methylene chloride / methanol = 10: 1). Crystallization from hexane monoethyl ether gave 103 mg of the title compound as a white crystalline solid.

Melting point 1 1 5 — 1 1 8

Elemental analysis: as the _{_{_{_{C 33 H 40 N 5 0 6}}}} C1 · H 2 0 ( molecular weight 6 ⁵ 6.19)

Theoretical value: C.60.40: H.6.45; N,〗 0.67: C1, 5.40

Found: C, 60.77: H, 6.56; N.10.62; C and 5.94.

M a s s: 638 (M + H) +.

NMR: δ (CDC13-270MHz): 8.88-8.79 (1Η, m), 8.50-8.14 (2Η, m), 7.94-7.64 (3Η.m) .7.21-6.72 (6Η.m), 6.26 ( 1Ή, br.s), 4.57-4.11 (7Η, m) .3.93-3.68 (3Η, m), 2.80-2.56 (5H, m)

(Example 8)

2 (S), 3 (S) -13- [N- (quinoxaline-12-carbonyl) -10-carboxymethoxyacetyl: L-threonyl 1-amino-2-hydroxy-4-phenylbutanol 4 _ (S) -clo!! -L-proline tert-butylamide (Exemplary Compound No. 7)

1) 2 (S), 3 (S)-3-[N-(quinoxaline-1-carbonyl)-0_-_ force) * _ _ _ _ tocinacetyl-L-threonyl] _ amino 2- 4- (7) -N-butadiene 4- (S) 1-L 1-L-proline tert-butylamide

Compound 5 synthesized in Example 5 (2) 2 (S) 3 (S) —3— [N— (quinoxaline-12-carbonyl) -1-L-threonyl] amino-2-methoxymethoxyloxy Phenylbutanol (4 (S) 1-mouth) Dissolve 1.20 g (0.18 mmol) of 1 L-proline tert-butylamide in 2 ml of methylene chloride, add 31 mg (0.27 mmol) of diglycolic anhydride and add Then, 2 mg (0.018 mmol) of 4-dimethylaminopyridine was added and the mixture was stirred at room temperature for 1 hour. The reaction mixture was washed with 5% citrate, 5% NaHCl3 aqueous solution and brine, and dried over anhydrous Na2S04. The solvent was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (methylene chloride / methanol / acetic acid = 90: 9: 3) to obtain 52 mg of the title compound.

M a s s: 799 (M + H) +.

NMR: δ (CDCI 3.270 MHz): 9.58 (1H, s), 8.64 (1Η, d, J = 7.9 Hz), 8.1 6-8.15 (2H, m), 7.89-7.84 (3H m), 7.27-7.12 ( 4H, m), 7.05-7.00 (1H, m), 6.62 (1H, br), 5.49-5.45 (1H, m), 4.91 -4.80 (1H, m) .4.72-4.61 (3H m), 4.47 (1H , t, J = 7.3Hz), 4.35 (1H, t, J = 5.9Hz). 4.29-4.13 (3H.m), 4.07 (2H.br.s). 3.70-3.63 (lH, m), 3.38- 3.29 (3H, m) ₍ 2.91-2.88 (2H, m), 2.64-2.47 (2H, m), 1.34-1.26 (12H, m).

2) 2 (S) 3 (S)-3--N-(quinoxaline-1-carbonyl) 1-Carboxymethoxyacetyl-L-threonyl 1_Amino 2-Hydrox-l-l-Phenylbutanoyl [4 (S) -Cross-lout 1 1: L-Proline tert-Butylamide

2 (S), 3 (S) -13- [N- (quinoxaline-12-carbonyl) -10-carboxymethoxyacetyl-l-threonyl] amino-2-methoxymethyloxy-14-phenylbutano [[4 (S) 1-mouth]-L-mouth mouth tert-Butylamide 40 mg (0.05 mmol)

In addition to 2 ml of 20% trifluoroacetic acid methylene chloride, the mixture was stirred at low temperature. The solvent was distilled off from the reaction mixture under reduced pressure, and benzene was added to remove trifluoroacetic acid as much as possible by azeotropic distillation, followed by crystallization with ether to obtain the title compound 21ing as a white crystalline solid. .

Elemental analysis: As C ₃₆ H ₄ 3N60l0Cl / 5H ₂₀ (molecular weight 845.28)

Theoretical: C, 51.15; H, 6.32; Ν, 9.94; C1, 4.19

Found: 51.48; H, 5.47: N, 9.29; C 1,4.29.

Mass: 755 (+ H) +.

NMR: δ (CDCl3, 270 MHz): 9.61 (1Η, s), 9.59 (lH, s), 8.70-8.66 (1 H, m), 8.27-8.19 (3H, m), 7.96-7.88 (3H.m) , 7.26-6.99 (5H.m), 6.80 -6.55 (1H, ra), 5.62 (1H, br), 5.08-5.05 (1H, br), .68-4.57 (2H, m) .4.41-4.15 (6H , m), 3.96-3.93 (1H, m), 2.84 (3H, br), 2.42-2.39 (1H.m), 1.35-1.22 (12H, m).

(Example 9)

2 (S), 3 (S) -13- [N- (quinoxaline-1-2-carbonyl) -1 0- (3-sulfoxybutanol) -1L-threonyl] amino1-2-hydr Roxy 4-phenylphenylbutanol [4 (S) -chloro mouth] 1 L-proline tert-butylamide (Exemplary Compound No. 8)

1) 2 (S), 3 (S) 13-[N-(Quinoxaline 1-2-carbonyl)-0-(3-carboxybutanol)-1-L-threonyl] _ amino 2 — Methoxymethyloxy 4-phenylbutanol 4- (S) 1-L-proline tert-Butylamide

2 (S), 3 (S) -13- [N- (quinoxaline-12-carbonyl) -1L-threonyl] amino 2-methoxymethyloxy 4-phenylbutyryl [4 (S ) One mouth] Dissolve 1.20 g (0.18 mmol) of 1 L-proline tert-butyl amide in 2 ml of methylene chloride, add 103 mg (0.9 mmol) of glutaric anhydride, and then add Dimethylaminopyridine 10 mg (0.09 mimol) was added, and the mixture was searched at room temperature for 1 hour. The reaction mixture 5% Kuen acid aqueous solution was washed successively with saturated brine and dried over anhydrous N a ₂ S 0 _4. The solvent was concentrated under reduced pressure, and the remaining brew was purified by silica gel chromatography (methylene chloride methanol = 10: 1) to obtain 93 mg of the title compound.

Mass: 797 (M + H) +.

NMR: δ (CDC13.270 MHz): 9.65 (lH, m), 8.59-8.56 (1H, m), 8.24- 8.19 (2H, m), 7.92-7.88 (2H, m), 7.26-7.00 (5H, m ), 6.40 (lH, br.s), 5.45-5.41 (lH, m), 4.79-4.61 (4H, m), 4.60-.20 (4H, m), 3.39-3.32 (3H, m), 2.88 ( 2H, d, J = 7.3Hz), 2.59 (2H, t, J = 7.3Hz), 2.44-2.33 (6H, m), 2.00-1.87 (3H, m), 1.37, 1.33 (9H, s, s) , 1.27 (3H, d, J = 6.6 Hz). 2) 2 (S), 3 (S) 1-3-[N-(quinoxalin-1-carbonyl) 1-1-(3-carboxybutanol) ) 1-L-threonyl] amino 1-2-hydroxy-4—phenylbutyryl “_4 (S) -chloroπ] —L-proline tert-butylamide

2 (S), 3 (S) -13- [N- (quinoxaline-12-carbonyl) -10- (3-carboxybutanoyl) -1-L-threonyl] amino-2-meth Xymethyloxy 4—phenylbutyrene [4 (S) —Cross mouth] —L—Proline tert—Butylamide 78 mg (0.098 mmol) in 20% trifluoracetic acid Z methylene chloride 5 ml In addition, the mixture was stirred at room temperature. The solvent is distilled off from the reaction mixture under reduced pressure, benzene is added, and azeotropic distillation is carried out as much as possible. After removing chloroacetic acid, the residue was crystallized from ether to obtain 36 mg of the title compound as a white crystalline solid.

Elemental _{_{analysis: C 37 H 4 5N609C1 / Η}} 2 0 ( molecular weight § § 1.2 ⁴⁾ and to

Theoretical: 57.62; Η, 6.14; 10., 10.90; C1, 4.60

Found: 57.65; H, 6.30; N, 10.57; C 1,4.54.

Mass s: 753 (M + H) +.

NMR: δ (CDC13, 270MHz): 9.61 (lH, s), 9.61 (1H, s), 8.62-8.59 (1H, m), 8.25-8.22 (2H, m), 7.96-7.91 (2H, m), 7.27-6.98 (4H, m), 6.48-6.41 (lH, m), 5.54-5.47 (lH, m), 4.90-4.85 (1H, m), 4.57-4.47 (2H, m), 4.39-4.24 (2H, m), 3.89-3.83 (1H, m), 2.87-2.65 (2H, m), 2.57-2.35 (7H, m), 2.02-1.89 (3H, m) ₍ 1.36-1.28 (9H, m) .

(Reference example 1)

2 (S), 3 (S) -1-3- (N-benzyloxycarbonyl) amino

2-Hydroxy 4-phenylbutyryl [4 (S) 1-close] 1-L-proline tert-butylamide

3 (S) —Amino 2 (S) —Hydroxy 4-phenylbutanol

4 (S) —Cross mouth]-Dissolve 1.5 g (3.93 mimol) of L-proline tert-butyl amide in 25 ml of methylene chloride and add N-carbobenzoxysuccinic acid imide. 1.47 g (5.90 mimol) and 0.55 ml (3.93 mimol) of triethylamine were added and stirred at room temperature for 2 hours. The reaction mixture 5% Kuen acid, and washed净with 5% N a HC O3 solution and saturated brine, and dried over anhydrous N a 2 S 0 _4. The solvent was concentrated under reduced pressure, and the oily residue was purified by silica gel column chromatography (methylene chloride Z-methanol) to obtain 1.9 g of the title compound as a colorless crystalline solid with hexane. Was.

NMR: δ (CDC13, 270MHz): 7.33-7.18 (10H, m), 6.29 (1Η, br), 5.26 (1Η, m), 5.02 (2H, m), 4.57-4.ll (4H, m), 3.87-3, 80 (1Η, m), 3.70 (1Η, d, J = 6.6), 2.71-2.58 (4Η, m), 1.29 (9Η.s). (Reference example 2)

2 (S), 3 (S) -13- (N-benzyloxycarbonyl) amino-2-tert-butyldimension) silyloxy_4 -_7 enylbutanoyl J4 (S) -chloro Mouth] _ One L One Proline

e t-butylamide

1.45 g (2.81 mimol) of the compound obtained in Reference Example 1 was dissolved in 15 ml of DMF, and 0.85 g (5.62 mimol) of tert-butyldimethylsilyl chloride and 0.30 g (5.62 mi) of imidazole were dissolved. ) And stirred at room temperature for 1 晚. Anti応混compound solvent was evaporated under reduced pressure, acetic Echiru 5 0 m 1 was added 5% Kuen acid, and washed诤with 5% N a HC 03 solution and saturated brine, anhydrous N a 2 S 0 ₄ It was dried. The solvent was concentrated under reduced pressure to obtain the title compound l. Llg.

NMR: δ (CDCl3, 270 MHz): 7.33-7.17 (10H, m), 6.60 (1H, br.s), 4.96 (2H, q, J = 35.0, 12.5 Hz), 4.87 (1H, d, J = 9.2Hz), 4.37-4.27 (3H, m), 4.10-3.99 (2H, m),

3.68 (lH, m), 3.12 (1H, dd, J = 14.5, 4.0Hz), 2.69-2.59 (2H, m), 2.34-2.26 (lH, m), 1.30 (9H, s), 0.93 ( 9H, s), 0.09 (6H, d, J = 4.6Hz)

(Reference example 3)

2 (S), 3 (S) -13- (N-benzyloxycarbonyl) amino 2-methoxymethyloxy 4-phenylbutanol [4 (S) -cyclo π 3-1 L -_7 ° π-line tert-butylamide

Reference Example] 1.95 g (3.78 mimol) of the compound obtained in was dissolved in DMF 4 O m 1, and N, N-diisopropylpyrethylamine 2 · 62 ml (15.12 mimol), and octamethylmethylether were dissolved; 1.20 ml (15.12 mmol) and 52 mg (0.43 mmol) of 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 3 days The solvent was distilled off from the reaction mixture under reduced pressure, and 50 ml of ethyl acetate was removed. Was added, and the mixture was washed with 5% citrate, 5% NaHC03 aqueous solution and saturated saline, and dried over anhydrous Na2S0. The solvent was concentrated under reduced pressure, and the oily residue was purified by silica gel chromatography.ー (Methylene chloride = 10: 1) to give 1.91 g of the title compound as a white crystalline solid.

Mp 7 1-7 3.

Elemental analysis: as the _{_{_{_{C 29 H 3 8N 3 0 6}}}} C1 ( molecular weight 560.10)

Theory: 62.19; H, 6.84; N, 7.50; C1, 6.33

Found: 61.63; H, 6.81; N, 7.31; C1.6.28.

Mass: 560 () ⁺ .

NMR: δ (CDC13, 270MHz): 7.38-7.18 (5H, s), 6.41 (1H, br.s), 5.39 (1H, s), 5.19 (2H, q, J = 4.2, 8.6Hz), 4.75- 4.63 (2H, m), 4.39-

4.14 (4H.m), 3.77-3.71 (1H, m), 3.33 (3H, s), 2.92 (2H, d, J = 7.3Hz), 2.72-2.65 (2H, m), 2.50-2.44 (2H, m), 1.61 (4H, s), 1.30 (9H, s).

1.04 (1H, d, J = 6.2Hz).

(Formulation example)

(Formulation example 1) (Hard capsule)

By filling each of the standard bisected hard gelatin capsules with 100 mg of the powdered compound of Example 1, 150 mg of lactose, 50 mg of cellulose and 6 mg of magnesium stearate, Manufacture unit capsules, wash and dry.

(Formulation Example 2) (Soft capsule)

A mixture of the compound of Example 1 in a digestible oil, for example, soybean oil, cottonseed oil, or olive oil, is prepared and injected into gelatin with a positive displacement pump to give a soft powder containing 100 mg of active ingredient. To obtain capsules, wash and dry. (Formulation Example 3) (Tablets)

In a conventional manner, 100 mg of the compound of Example), 0.2 mg of co-acid silicon dioxide, 5 mg of magnesium stearate, 275 mg of microcrystalline cellulose, 11 mg of starch and 98.8 mg of It is manufactured using lactose.

In addition, a coat is applied as required. (Formulation Example 4) (Injection)

1.5% by weight of the compound of Example 1 is stirred in 10% by volume of propylene glycol, made up to volume with water for injection, and then sterilized to produce.

(Formulation Example 5) (Suspension)

In 5 ml, 100 mg of the finely divided compound of Example 1, 100 mg of sodium carboxymethylcellulose, 5 mg of sodium benzoate, 1.0 g of sorbitol solution (Japanese Pharmacopoeia) and Produced to contain 0.025 ml of vanillin.

(Test Example 1)

Measurement of HIV protease inhibitory activity

Using the HIV protease expressed in Escherichia coli and a substrate synthesized, the HIV protease inhibitory activity of the peptide derivative of the present invention was determined using the IC ₅₀ value as an index as follows.

a) Creation of expression vector

From the BH10 clone containing the main part of the HT LVIIIB provirus (Flossie Wong-Staal 'et al., Nature, vol. 313, 277-284, 19S5), C1 in the virus gag region The region between the aI site and the Ec0RI site in the po1 region was cut out, and the obtained fragment was cloned into the same site of pBR322.

Next, a synthetic fragment containing a translation initiation codon (ATG) was inserted between the BamHI and C1aI sites 5 'upstream of the insertion site.

And a fragment consisting of the nucleotide sequence of T7 Promoter region [(BglII to BamHI fragment) Barbara A. Moffatt et al., J. Mol. Biol., 189 Vol., Pp. 113-130, described in 1986]. After cutting the Bg1II site (uppermost region of the po1 region) in the obtained plasmid, repair synthesis was performed using klenow fragment, and then T4DNA ligation was performed. And recombined to construct an expression vector pT7HIV.GP (-) containing the HIV gag region and a part of the p01 region. b) Expression in E. coli

Escherichia coli BL-21 containing the T7 polymerase gene [(DE-3) Barbar a A. Mofiatt et al., J. Mol. Biol., 189, 113-130, 1986] describes: After transfection of pT7HIV.GP (-), the resulting transformants were transformed into 600% of M9CA—10% LB medium containing 200 ^ gZml of ampicillin. Culture at 37 until the absorbance at nm reaches 2.

0.4 mM of isopropylthio-1- / D-galactoside was added, and the culture was further continued for 3 hours.

The obtained cells were collected and stored as pellets at 18. The polyprotein synthesized from the expression vector is degraded in the cells by autolysis, and the HIV protease itself is produced.

c) Purification

Cells obtained from 2 liters of the culture solution were mixed with 60 ml of Knuffer A (50 mM Tris-HCl (ρΗ7.5), ImM dithiothreitol, 0.7 mL). % Lysozyme,] 0 g / m1 aprotinin, 5 mM ethylenediaminetetraacetic acid, 10 / g / ml benzamide,] mM fluorinated phenylmethylsulfonic acid, 10% glycerin) After turbidity and left at 0 for 10 minutes, Triton

X-100 (0.1%) was added, and the mixture was left at 0 for 10 minutes.

After freeze-thawing four times, DNaseI (0.1 mg) and 10 mM magnesium chloride were added to decompose DNA in the suspension.

The supernatant obtained by centrifuging this suspension was applied to DEAE Sephade XA25 column (5 cm inner diameter X 20 cm) to fractionate. The column is a buffer B [5 O mMHE PES (pH 7.8),] mM dithiol. Plate, lO ^ gZmI aprotinin, 5 mM ethylenediaminetetraacetic acid, 1 OgZml benzamide, 1 mM fluormethylsulfonic acid, 10% glycerin] Using.

The flow-through fractions with recognized activity were collected, precipitated with ammonium sulfate (60%), and the resulting precipitate was washed with 2 ml of buffer C (50 mM Tris-HCl (pH 7.5), Dissolve in ImM dithiothreitol, ImM eradiamine tetraacetic acid, 200 mM sodium chloride) and use a TSK. S200 SW SW gel filtration column (7.5 mm ID x 60 cm, The product was fractionated at a flow rate of 0.5 m1Z using Knuffer C.

The obtained active fraction was concentrated twice by using an ultrafiltration membrane having a molecular weight cut off of 10,000 daltons, and stored at 180 as an enzyme solution.

d) Activity measurement

The measurement of the HIV protease inhibitory activity was carried out based on the method of E.D.Matey0shhi et al. (Science 247,954 (1990)).

That is, a synthetic substrate of 20 4M 4- (4-dimethylaminophenylazo) benzoic acid (DABCY L) -Ser-Gln-Asn—Tyr—Pro—

I 1 e-V a 1 G in— 5 — [(2-Aminoethyl) amino] naphthalene: I-sulfonic acid (EDAN S) (purchased from BAC HEM), dissolved in DMS O A reaction solution of 50 mM sodium acetate (pH 5.5) -11 M saline containing the inhibitor at each concentration was prepared. A partially purified recombinant HIV protease was added thereto, the reaction was started at room temperature, and the reaction product was measured by a fluorescence spectrophotometer. The amount of reaction produced per unit time in the presence of each concentration of the inhibitor was calculated, and the 509 inhibition degree (IC ₅₀ ) was determined.

The compounds of the present invention showed strong inhibitory activity.

(Test Example 2)

HI ^ Some. Inhibition of virus release from EM cells (Method)

HIV (HT LVIIIB) persistently infected cells Mo1t4 cells (Mo1t4ZHTLVIIIB) were transformed twice using RPMI1640 medium (containing 1096 non-immobilized fetal calf serum). After washing, the cells were inoculated into the same medium at a concentration of ixi 05 cells, and cultured at 37 under 5% carbon dioxide gas. After 2 hours, the culture supernatant was obtained by centrifugation and used as virus stock: CΕΜ cells were grown in RΡΜI 1640 medium (containing 10% non-immobilized fetal calf serum) for 2 hours. xi 0 was adjusted to 5 cells ^/ / ml, were added to the virus scan bets get diluted to a suitable concentration. Further, various concentrations of the compounds were added, and the cells were cultured under 37: 5% carbon dioxide. Seven days later, the culture supernatant was collected, and the amount of HIV antigen contained therein was measured by the EIA method (HIV antigen, EIAII, Abbott), and the release of the compound was suppressed by 90% by adding the compound. Io is called I Cgo.

The compounds of the present invention exhibited strong virus release inhibitory activity.

(Test Example 3)

The compounds of the present invention exhibit pharmacodynamic properties which are expected to exert the above inhibitory effects in vivo. For example, when 1 OmgZkg of the compound of the present invention is intravenously administered to a rat, the blood level at 1 hour after the administration is almost the same as or higher than the ED go value in the cell test. Similarly, in the case of intraduodenal administration or oral administration of the compound 30 mg / kg, the concentration detected in the blood of the rat 30 minutes after administration is higher than the ED90 value in the cell test, preferably the cell concentration. It is about 6 times the ED 90 value in the test.

To measure blood levels, prepare a solution for administration as follows. For intravenous administration, 1 Omg of the test compound is dissolved in 0.3 ml of dimethylacetamide (DMA), and a solution is prepared by adding 0.4 ml of PEG 400 ml and 0.3 ml of water I do. For intraduodenal and oral administration, 30 m of the test compound was dissolved in 0.6 ml of DMA, then 0.8 ml of PEG and 0.8 ml of water. Prepare a solution containing 6 ml. After administration of these solutions to the rat, blood is collected over time, and the blood is centrifuged (300 rpm, 10 minutes) to obtain plasma. Analyze by HPLC using the column switching method to determine the plasma concentration. Column switching HP LC conditions were as follows: 1 column, 1 column ^f , 300 mm length, 4.6 mm diameter, TSKG el G-200 O SW (manufactured by Tosoh Corporation). % Phosphoric acid (1: 9 VZ V%) is used as eluent. Column 2 uses the same eluent as column 1 on a YMC A-312 ODS (YMC) with a length of 50 mm and a diameter of 6 mm. However, the content of acetonitrile should be 40-70% depending on the test compound. Effect of the Invention The compound of the present invention has excellent and specific anti-HIV protease inhibitory activity, and also has an excellent inhibitory effect on virus release from HIV-infected cells. Furthermore, the compounds of the present invention exhibited excellent oral absorption and high blood levels in living organisms.

Claims

Claims General formula

[In the formula, R ¹ represents a quinolin-12-carbonyl group or a quinoxaline-12-carbonyl group, and R2 represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, an acetyl group, a methoxymethyl group. Group, methylthiomethyl group, acetomethyl group, vivaloyoxymethyl group, H〇〇C (C H2) _n C group (n is an integer of 2 to 4) or H 00 CC H2 0 CH ₂ CO —Indicating a group] and salts thereof.

2. The compound according to claim 1, wherein R 2 is a hydrogen atom or an alkyl group having 1 to 4 carbon atoms.

3. The compound according to claim 1, wherein R2 is a hydrogen atom.

4.2 (S), 3 (S) -1-3- [N- (quinoxaline-12-carbonyl) -1-L-threonyl] amino-2—hydroxy-4-phenylbutanol-[4 (S) 1-channel] 1-L-proline tert-butylamide.

5.2 (S), 3 (S)-3-[N- (quinoline-l-carbonyl) -l-L-threonyl] amino-2-hydroxyl 4-phenylbutanol B) L-proline tert-butylamide.

6. A protease inhibitor comprising an effective amount of the compound selected from claim 1 and a pharmacologically acceptable carrier or excipient.

7. Valid! A prophylactic or therapeutic agent for HIV infection comprising a compound selected from claim 1 and a pharmacologically acceptable carrier or excipient.

8. A protease inhibitor comprising an effective amount of the compound of claim 2 and a pharmacologically acceptable carrier or excipient.

9. A prophylactic or therapeutic agent for HIV infection comprising an effective amount of the compound of claim 2 and a pharmacologically acceptable carrier or excipient.

10. A protease inhibitor comprising an effective amount of the compound of claim 3 and a pharmacologically acceptable carrier or excipient.

1 1. A prophylactic or therapeutic agent for HIV infection comprising an effective amount of the compound of claim 3 and a pharmacologically acceptable carrier or excipient.

1 2. A protease inhibitor comprising an effective amount of the compound selected from claim 4 and a pharmacologically acceptable carrier or excipient.

1 3. A prophylactic or therapeutic agent for HIV infection comprising an effective amount of the compound selected from claim 4 and a pharmacologically acceptable carrier or excipient.

14. A protease inhibitor comprising an effective amount of the compound selected from claim 5 and a pharmacologically acceptable carrier or excipient.

15. A prophylactic or therapeutic agent for HIV infection comprising an effective amount of the compound selected from claim 5 and a pharmacologically acceptable carrier or excipient.