JPH10147566A - Vascularization-inhibiting medicine - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the new medicine useful as a medicine for inhibiting vascularization occurring in vulnerary, inflammation, tumor growth, diabetic retinopathy, immature infant retinopathy, retinal venous obstruction, senile disciform degeneration of macula, etc. SOLUTION: This vascularization-inhibiting agent contains a calpain-inhibiting compound of formula I or II [R<1> is carboxyl, ethoxycarbonyl, etc.; R<2> is H or a lower alkyl; R<3> is benzyl, isobutyl, etc.; R<4> is H, etc., or R<4> is combined with R<2> to form a ring; R<5> is (substituted) phenyl; R<11> is naphthyl, (substituted) phenyl; R<12> , R<13> are each isopropyl, tertiary butyl, etc.; R<14> is benzyl, indol-2- ylmethyl, etc.; (n) is 1, 0]. The compound of formula I is obtained by reacting a compound of formula III (including its reactive derivative or its salt) with a compound of formula IV (including its reactive derivative or its salt).

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an angiogenesis inhibitor comprising a cysteine protease inhibitory compound.

[0002]

2. Description of the Related Art Angiogenesis is the formation of new blood vessels in a living body,
A phenomenon in which a new vascular network is formed. Angiogenesis is observed in embryos, fetuses, placenta, uterus, etc. under normal physiological conditions such as development and reproduction, when it is associated with wound healing, inflammation, tumor growth, etc. Is a pathological phenomenon seen in diabetic retinopathy, retinopathy of prematurity, retinal vein occlusion, and senile discoid macular degeneration.

[0003] Angiogenesis largely depends on the function and proliferation of endothelial cells, and is considered to be a cascade reaction that proceeds in the smallest vein according to the following process. That is,
(1) activation of differentiated resting vascular endothelial cells, (2) disruption of cell matrix such as basement membrane by endothelial cells expressing protease activity, (3) migration of endothelial cells, (4) endothelial cells It is thought that new blood vessels are formed by the continuous occurrence of elementary reactions such as proliferation of endothelial cells and (5) vascularization of endothelial cells due to differentiation [T. Oik
awa, Drug News Perspest, Vol. 6, pp. 157-162 (1
993)]. It has been found that each of these processes is promoted by pro-angiogenic factors. These angiogenesis-promoting factors include tumor angiogenetic factor (TAF) secreted from tumor tissue and fibroblast growth factor (FG) present in various normal tissues.
F), growth factors such as platelet-derived endothelial cell growth factor and vascular endothelial cell growth factor. In addition, it has been reported that cytokines, prostaglandins, monobutyrin, angiogenin and the like have similar effects directly or indirectly (M. Klagsbrun et al., Annu. Rev. Physio
l., 53, 217-239 (1991)].

[0004] Substances that suppress such angiogenesis include:
Angiostatic steroids such as cortisone that inhibit endothelial cell proliferation [Folkman, J. et al., Scie
nce, vol.221, p.719 (1983)], medroxyprogesterone acetate which inhibits plasminogen activator production by endothelial cells, fumagillinic acid derivative which inhibits endothelial cell growth inhibition and tube formation, endothelial cell proliferation And SD-4152, a sulfate polysaccharide that inhibits migration, and retinoic acid that is involved in the modification of endothelial cell differentiation (Tsushi Oikawa, Vascular and Endothelial, 2, 470-478, 1992).
Year).

[0005]

However, the above-mentioned drug for suppressing angiogenesis is a therapeutic agent which suppresses angiogenesis clinically, for example, when there are strong side effects and there is a problem in safety, or the effect is not sufficient. It is not yet established.

[0006]

Means for Solving the Problems The present inventors have conducted intensive studies with the aim of developing a drug that strongly suppresses angiogenesis. As a result, they have found that a compound having a cysteine protease inhibitory activity has a strong angiogenesis inhibitory effect, and completed the present invention. That is, according to the present invention,
(1) An angiogenesis inhibitor comprising a cysteine protease inhibitory compound is provided.

[0007] Cysteine protease refers to a protease having a cysteine residue in the active center of the enzyme among proteases. Representative examples thereof include lysosomal enzymes cathepsin B, H, L, dipeptidyl peptidase and cytoplasm. Calpain and the like. Although the physiological role of these enzymes is often unknown, their role has been gradually elucidated in recent years. For example, calpain is a proteolytic enzyme that exists widely in the body, is activated by calcium ions, and has an optimum pH near neutrality.
Its roles have been elucidated to date, such as the degradation of cytoskeletal proteins, the activation of inactive cell precursors such as protein kinase C, and the degradation of receptor proteins.
Furthermore, it has been revealed that abnormalities in the activity of this enzyme are involved in many diseases. For example, involvement in intractable diseases such as stroke, subarachnoid hemorrhage, Alzheimer's disease, ischemic disease, muscular dystrophy, cataract, platelet aggregation, arthritis, and osteoporosis has been suggested (Trends in Pharmaco
logical Science, vol. 15, p. 412 (1994)].

Further, the following embodiments can be mentioned as the angiogenesis inhibitor of the present invention. (2) The angiogenesis inhibitor according to the above (1), wherein the cysteine protease inhibitory compound is a calpain inhibitory compound.

(3) The calpain inhibitory compound is selected from calpastatin and calpastatin peptide.
The angiogenesis inhibitor according to the above (2), which is at least one compound.

(4) The calpastatin peptide has the following general formula: -Gly-A-Tyr-Arg- (where A is -Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys
-, -Lys-Arg-Glu-Val-Thr-Leu-Pro-Pro-Lys-, -Glu-As
p-Asp-Glu-Thr-Ile-Pro-Ser-Glu-, -Glu-Asp-Asp-Glu-
Thr-Val-Pro-Pro-Glu-, -Glu-Asp-Asp-Glu-Thr-Val-Pr
o-Ala-Glu-, -Glu-Lys-Glu-Glu-Thr-Ile-Pro-Pro-Asp-
Or -Glu-Arg-Asp-Asp-Thr-Ile-Pro-Pro-Glu-. (3) The angiogenesis inhibitor according to the above (3), which is one or more compounds selected from peptides having the amino acid sequence of (3).

(5) The angiogenesis inhibitor according to the above (4), wherein the calpastatin peptide has the following amino acid sequence: Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Ly
s-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-
Leu-Ala

(6) The angiogenesis inhibitor according to the above (2), wherein the calpain inhibitory compound is at least one compound selected from substances which inhibit a Ca ²⁺ binding site having high homology to calmodulin of calpain.

(7) The angiogenesis inhibitor according to the above (6), wherein the substance that inhibits a Ca ²⁺ binding site having high homology to calmodulin of calpain is at least one compound selected from calmodulin antagonistic compounds.

(8) The cysteine protease inhibitory compound is selected from epoxy succinate peptide compounds, peptide aldehyde compounds, peptide halomethane compounds, peptide diazomethane compounds, peptide halohydrazide compounds, peptide disulfide compounds, peptide ketoamide compounds and isocoumarin compounds. The angiogenesis inhibitor according to claim 1, which is one or more compounds.

(9) The cysteine protease-inhibiting compound is an epoxysuccinate peptide compound (8).
An angiogenesis inhibitor according to the above.

(10) An epoxysuccinic acid peptide compound represented by the formula (I):

[0017]

Embedded image

[In the formula, R ¹ represents a carboxyl group which may be esterified or a carboxamide which may be substituted, and R ² represents hydrogen or lower (unless otherwise specified in the present specification, “lower”). May represent an alkyl group or may be linked to R ³ or R ⁴ to form a ring, wherein R ³ and R ⁴ are the same or different and are hydrogen, substituted It is a lower alkyl group or a substituted or show a good sulfide group, also R ³
And R ⁴ may be linked to form a ring, and R ⁵ is a group represented by the formula (I
I)

[0019]

Embedded image

Wherein R ⁶ represents a halogen atom or an alkoxy group, or a substituted phenyl group represented by the formula (III): SO ₂ -R ⁷ (III) (wherein R ⁷ is substituted with a lower alkyl group) (Indicating an optionally substituted aryl group or an optionally substituted amino group), and n represents 0 or 1.] or a salt thereof. Angiogenesis inhibitors.

(11) The angiogenesis inhibitor according to the above (10), wherein R ¹ is a carboxyl group which may be esterified, or a carboxamide group which may be substituted with a hydroxy group or an aralkyloxy group.

(12) The angiogenesis inhibitor according to the above (10), wherein R ² is hydrogen or a methyl group.

(13) The angiogenesis inhibitor according to the above (10), wherein R ² is linked to R ³ or R ⁴ to form a pyrrolidine ring.

(14) R ³ and R ⁴ are the same or different and are sulfide groups which may be substituted with hydrogen, aromatic or lower alkyl groups which may be substituted with a carbamoyl group or acylamino groups. )).

(15) The angiogenesis inhibitor according to the above (10), wherein R ³ and R ⁴ are linked to form a cyclopentane ring.

(16) The angiogenesis inhibitor according to the above (10), wherein R ^{6 in the} formula (II) is chlorine or fluorine.

(17) The angiogenesis inhibitor according to the above (10), wherein R ^{7 in the} formula (III) is a phenyl group or a dimethylamino group which may be substituted with a lower alkyl group.

(18) The angiogenesis inhibitor according to the above (8), wherein the cysteine protease inhibitory compound is a peptide aldehyde compound.

(19) The angiogenesis inhibitor according to the above (18), wherein the peptide aldehyde compound is leupeptin.

(20) The peptide aldehyde compound represented by the formula (VI):

[0031]

Embedded image

(In the formula, R ¹¹ represents an aryl group having 6 to 10 carbon atoms which may have a substituent, and R ¹² and R ¹³ may be the same or different and may be hydrogen, an alkyl group having 1 to 4 carbon atoms. Or may be linked to form a ring having 3 to 7 carbon atoms, and R ¹⁴ represents a lower alkyl group optionally substituted with an aryl group, a cycloalkyl group or an aromatic heterocyclic ring)
19. The angiogenesis inhibitor according to claim 18, which is a compound represented by the formula: or a salt thereof.

(21) The angiogenesis inhibitor according to the above (20), wherein R ¹¹ is a phenyl group or a naphthyl group which may be substituted with fluorine, chlorine or methyl group.

(22) R ¹¹ is 4-fluorophenyl,
The angiogenesis inhibitor according to the above (21), which is a group selected from 4-chlorophenyl, p-tolyl and 2-naphthyl.

(23) The angiogenesis inhibitor according to the above (20), wherein R ¹² is propyl, isopropyl or tert-butyl and R ¹³ is hydrogen.

(24) R ¹² is isopropyl;
^The angiogenesis inhibitor according to the above (23), wherein ¹³ is hydrogen.

(25) The angiogenesis inhibitor according to the above (20), wherein the ring formed by linking R ¹² and R ¹³ is cyclohexylidene.

(26) R ¹⁴ is isobutyl, benzyl,
The angiogenesis inhibitor according to the above (20), which is cyclohexylmethyl or indol-3-ylmethyl.

(27) Formula (VI):

[0040]

Embedded image

(In the formula, R ¹¹ represents an aryl group having 6 to 10 carbon atoms which may have a substituent, and R ¹² and R ¹³ may be the same or different, and may be hydrogen, an alkyl group having 1 to 4 carbon atoms. Or may be linked to form a ring having 3 to 7 carbon atoms, and R ¹⁴ represents a lower alkyl group optionally substituted with an aryl group, a cycloalkyl group or an aromatic heterocyclic ring)
Or a salt thereof.

(28) The compound of the above-mentioned (27), wherein R ¹¹ is a phenyl group or a naphthyl group optionally substituted with fluorine, chlorine or methyl group, or a salt thereof.

(29) The compound or a salt thereof according to the above (27), wherein R ¹¹ is a group selected from 4-fluorophenyl, 4-chlorophenyl, p-tolyl and 2-naphthyl.

(30) The compound of the above-mentioned (27), wherein R ¹² is propyl, isopropyl or tert-butyl and R ¹³ is hydrogen, or a salt thereof.

(31) R ¹² is isopropyl,
The compound or a salt thereof according to the above (27), wherein R ¹³ is hydrogen.

(32) The compound or a salt thereof according to the above (27), wherein the ring formed by linking R ¹² and R ¹³ is cyclohexylidene.

(33) The compound or a salt thereof according to the above (27), wherein R ¹⁴ is isobutyl, benzyl, cyclohexylmethyl or indol-3-ylmethyl.

[0048]

BEST MODE FOR CARRYING OUT THE INVENTION As the cysteine protease inhibitor used in the angiogenesis inhibitor of the present invention, any compound that inhibits cysteine protease can be suitably used. As such a compound, for example, (+)-(2S, 3S) -3-[[[1-
[[[4-[(aminoiminomethyl) amino] butyl]
Amino] carbonyl] -3-methylbutyl] amino] carbonyl] -2-oxiranecarboxylic acid (E-64);
(+)-(2S, 3S) -3 [(S) -3-methyl-1
-(3-methylbutylcarbamoyl) butylcarbamoyl] -2-oxiranecarboxylic acid (E-64c),
(+)-(2S, 3S) -3 [(S) -3-methyl-1
-(3-methylbutylcarbamoyl) butylcarbamoyl] -2-oxiranecarboxylic acid ethyl ester (E-
Epoxy succinate peptide compounds such as 64d); leupeptin, calpeptin, Ac-Leu-Leu-nLeu-H (calpain inhibitor peptide I), Ac-Leu-Leu-nMet-H
(Calpain inhibitor peptide II), Z-Val-Phe-H
(MDL28170), peptide aldehyde compounds such as Boc-Leu-Nle-H; peptide halomethane compounds such as Z-Leu-Leu-Tyr-CH ₂ F; peptide diazomethane compounds such as Z-Leu-Leu-Tyr-CHN ₂ ; Z-3-I-Tyr -NHNHCOCH peptide halo hydrazide compounds such as ₂ I; Leu-Leu- (3- nitro-2-pyridine sulfenyl) peptide disulfide compounds such as _{Cys-NH 2; Z-Leu} -Abu- CONHEt (AK27
Peptide ketoamide compounds such as 5); 7-amino-4
And isocoumarin compounds such as -chloro-3- (3-isothioureidopropoxy) isocoumarin.

The cysteine protease inhibiting compound used in the angiogenesis inhibitor of the present invention is calpain or cathepsin B, H, L among cysteine proteases.
Can be used. Among them, examples of the substance that specifically inhibits calpain include calpastatin and calpastatin peptide.

The calpastatin peptide has the following general formula: -Gly-A-Tyr-Arg- (where A is -Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys
-, -Lys-Arg-Glu-Val-Thr-Leu-Pro-Pro-Lys-, -Glu-As
p-Asp-Glu-Thr-Ile-Pro-Ser-Glu-, -Glu-Asp-Asp-Glu-
Thr-Val-Pro-Pro-Glu-, -Glu-Asp-Asp-Glu-Thr-Val-Pr
o-Ala-Glu-, -Glu-Lys-Glu-Glu-Thr-Ile-Pro-Pro-Asp-
Or a peptide having the amino acid sequence of -Glu-Arg-Asp-Asp-Asp-Thr-Ile-Pro-Pro-Glu-), and particularly preferably a peptide having the following amino acid sequence (27-mer calpastatin peptide). Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Ly
s-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-
Leu-Ala

The cysteine protease inhibitory compound used in the angiogenesis inhibitor of the present invention also includes a substance that inhibits a Ca ²⁺ binding site having high homology to calmodulin of calpain. Such substances include melittin, calmidazolium, trifluoperazine and N-
Calmodulin antagonistic compounds such as (6-aminohexyl) -5-chloro-1-naphthalenesulfonamide hydrochloride (W7) and ethylenediaminetetraacetic acid (EDTA)
And so on. Any of these cysteine protease inhibiting compounds can be used alone, or two or more can be used in combination.

Further, as an epoxy succinate peptide compound having a strong cysteine protease inhibitory activity,
A novel compound represented by the following general formula (I) can be used.

[0053]

Embedded image

(Wherein each symbol is as defined above)

In the above formula (I), examples of the optionally esterified carboxyl group represented by R ¹ include a carboxyl group and an alkoxycarboxyl group. As the alkoxy group of the alkoxycarboxyl group, for example, an alkoxy group having 1 to 6 carbon atoms, preferably an alkoxy group having 1 to 4 carbon atoms, specifically, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec. -Butoxy and tert-butoxy. Of these, ethoxy is particularly preferred. Examples of the substituent of the optionally substituted carboxamide represented by R ¹ include a hydroxy group, an alkoxy group (eg, methoxy, ethoxy, propoxy) and an aralkyloxy group (eg, benzyloxy). Preferred are hydroxy and benzyloxy.

As the lower alkyl group represented by R ² ,
A linear or branched alkyl group having 1 to 6 carbon atoms, preferably an alkyl group having 1 to 4 carbon atoms, specifically, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec- Butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl, 4-methylpentyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl and the like. Preferably, they are hydrogen and methyl.

The ring formed by linking R ² to R ³ or R ⁴ includes, for example, aziridine, azetidine, pyrrolidine, piperidine and the like. Of these, pyrrolidine is particularly preferred.

As the alkyl group of the lower alkyl group which may be substituted for R ³ and R ⁴ , a linear or branched alkyl group having 1 to 6 carbon atoms, specifically, methyl,
Ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl,
4-methylpentyl, 1,1-dimethylbutyl, 2,2
-Dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl and the like. Preferred are methyl, ethyl, isobutyl and sec-butyl. Examples of the substituent which the alkyl group may have include an aromatic ring and a carbamoyl group. Examples of the aromatic ring include an aromatic carbon ring such as a benzene ring and an aromatic heterocyclic ring such as an indole ring. Of these, a benzene ring is particularly preferred.

The sulfide group of the sulfide group which may be substituted by R ³ or R ⁴ includes an alkylthioalkyl group, preferably a C ₁ -C ₄ alkylthio C ₁ -C ₄
Alkyl group, specifically, dimethyl sulfide, diethyl sulfide, dipropyl sulfide, dibutyl sulfide, dipentyl sulfide, dihexyl sulfide,
Methyl ethyl sulfide, methyl propyl sulfide,
Ethyl butyl sulfide and the like can be mentioned. Preferably, dimethyl sulfide and methyl ethyl sulfide are used. Examples of the substituent that the sulfide group may have include an acylamino group. Examples of the acylamino group include formylamino, acetylamino, propionylamino, butyrylamino, isobutyrylamino, valerylamino, isovalerylamino, pivaloylamino, and n-hexanoylamino. Preferably, it is acetylamino.

The ring which may be formed by linking R ³ and R ⁴ includes cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane and the like. Of these, cyclopentane is particularly preferred. Formula (I
Examples of the halogen atom represented by R ⁶ which is a substituent of the substituted phenyl group represented by I) include fluorine, chlorine, bromine and iodine. Preferred are fluorine and chlorine. These halogen atoms may be substituted at any of the meta, para and ortho positions of the phenyl group. As the alkoxy group of the substituent R ⁶ of the substituted phenyl group (II), an alkoxy group having 1 to 6 carbon atoms, preferably an alkoxy group having 1 to 4 carbon atoms, specifically, methoxy, ethoxy, propoxy, Isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-
Butoxy and the like. Of these, methoxy is particularly preferred.

Examples of the aryl group of the aryl group optionally substituted by lower alkyl represented by R ⁷ , which is a substituent of the substituted sulfonyl group represented by the formula (III), include a phenyl group and a naphthyl group. The lower alkyl group which may be substituted on the aryl group includes, for example, methyl, ethyl, propyl, isopropyl, butyl and the like. The substituent may be substituted at any position of the aryl group.

Examples of the amino group represented by R ⁷ include an amino group and a group in which one or two amino groups are substituted with a linear, branched or cyclic alkyl group having 1 to 6 carbon atoms, for example, Methylamino, dimethylamino, ethylamino, diethylamino, propylamino, dipropylamino, isopropylamino, diisopropylamino,
Butylamino, dibutylamino, cyclohexylamino and the like. Of these, dimethylamino is particularly preferred.

The salt of the compound represented by the formula (I) is preferably a physiologically acceptable salt, for example, a salt with an inorganic base, a salt with an organic base, a salt with an inorganic acid, or an organic acid. And salts with basic or acidic amino acids. Preferred examples of the salt with an inorganic base include, for example,
Alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt and magnesium salt;
And aluminum salts and ammonium salts. Preferred examples of the salt with an organic base include, for example,
Salts with trimethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, N, N-dibenzylethylenediamine and the like can be mentioned. Preferable examples of salts with inorganic acids include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Suitable examples of salts with organic acids include, for example, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, Salts with p-toluenesulfonic acid and the like can be mentioned. Preferred examples of the salt with a basic amino acid include, for example, salts with arginine, lysine, ornithine and the like. Preferred examples of the salt with an acidic amino acid include, for example, salts with aspartic acid, glutamic acid, and the like. Is mentioned. The compound represented by the general formula (I) is represented by the following reaction formula

[0064]

Embedded image

(Wherein each symbol is as defined above). In the present production method, a compound represented by the general formula (IV) or a reactive derivative or a salt thereof at the carboxyl group is represented by the compound represented by the general formula (V). [The compound (I) can be produced by reacting the compound [V] (hereinafter sometimes referred to as compound (V)) or a reactive derivative thereof or a salt thereof.

In the above production method, for example, a conventional means of peptide synthesis such as a liquid phase synthesis method and a solid phase synthesis method is used. Such a method of peptide synthesis may be in accordance with any known method. For example, Nobuo Izumiya et al., "Basic and Experimental Peptide Synthesis", Maruzen Co., Ltd., 1985; Haruaki Yajima, Shunpei Sakakibara, "Biochemical Experiment Lecture 1", edited by The Biochemical Society of Japan, Tokyo Kagaku Doujin, 1977; Toshiya Kimura, "Serial Chemistry Experimental Lecture 1", edited by the Biochemical Society of Japan, Tokyo Kagaku Dojin, 1987; Nobuo Suzuki, " 4th edition Experimental Chemistry Course 22 Organic Synthesis IV ”,
It is manufactured by the method described in The Chemical Society of Japan, Maruzen Co., Ltd., 1992, or a method similar thereto.

Suitable reactive derivatives at the carboxyl group of compound (IV) include acid halides, acid anhydrides, activated amides, activated esters and the like.
Acid halides include acid chlorides, and acid anhydrides include, for example, substituted phosphoric acids (such as dialkyl phosphoric acid, phenyl phosphoric acid, diphenyl phosphoric acid, dibenzyl phosphoric acid, and halogenated phosphoric acid), and dialkyl phosphoric acids. Phosphorous acid, sulfurous acid, thiosulfuric acid, sulfuric acid, sulfonic acid (such as methanesulfonic acid), aliphatic carboxylic acid (such as acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, pentanoic acid, isopentanoic acid, and trichloroacetic acid) or aromatic Mixed anhydrides with acids such as aromatic carboxylic acids (such as benzoic acid) or symmetrical acid anhydrides. Suitable examples of the activated amide include, for example, imidazole, 4-substituted imidazole, dimethylpyrazole, triazole or tetrazole. Preferred examples of the activated ester include, for example, cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl ester, vinyl ester, propargyl ester, p-nitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, methylphenyl ester, phenyl ester Azophenyl ester, phenylthioester, p-
Nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, 8-quinolyl thioester, or N, N-dimethylhydroxyamine, 1-hydroxy-2- (1H) -pyridone, N-hydroxy Esters with N-hydroxy compounds such as succinimide, N-hydroxyphthalimide, 1-hydroxy-1H-benzotriazole and the like.

Suitable salts of compound (IV) and its reactive derivative include, for example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, and aluminum salt.
Ammonium salts such as organic base salts such as trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, diethanolamine salt, triethanolamine salt, dicyclohexylamine salt and N, N-dibenzylethylenediamine salt; Base salts are mentioned. These reactive derivatives can be arbitrarily selected depending on the type of the compound (IV) to be used.

Suitable reactive derivatives at the amino group of compound (V) include Schiff base-type imino or enamine-type tautomers formed by the reaction of compound (V) with a carbonyl compound such as an aldehyde or ketone. ,
Alternatively, a silyl derivative formed by reacting compound (V) with a silyl compound such as bis (trimethylsilyl) acetamide, mono (trimethylsilyl) acetamide, bis (trimethylsilyl) urea, or compound (V) with phosphorus trichloride or phosgene And derivatives formed by the reaction with Suitable salts of compound (V) and its reactive derivatives include, for example, inorganic acid salts such as hydrochloride, hydrobromide, nitrate, sulfate and phosphate, for example, formate, acetate, triacetate and the like. Organics such as fluoroacetate, fumarate, oxalate, tartrate, maleate, citrate, succinate, malate, methanesulfonate, benzenesulfonate and p-toluenesulfonate Acid salts. These reactive derivatives can be arbitrarily selected depending on the type of compound (V) used.

The reaction between the compounds (IV) and (V) is usually carried out with water,
For example, alcohols such as methanol and ethanol,
The reaction is carried out in a common solvent such as acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N, N-dimethylformamide, pyridine, but any other solvent which does not adversely affect the reaction. The reaction can be carried out in any organic solvent. These conventional solvents may be used as a mixture with water.

In this reaction, when the compound (IV) is used in a free form or a salt thereof, N, N'-dicyclohexylcarbodiimide, N-cyclohexyl-
N'-morpholinoethylcarbodiimide, N-cyclohexyl-N '-(4-diethylaminocyclohexyl)
Carbodiimide, N, N′-diethylcarbodiimide,
N, N'-diisopropylcarbodiimide, N-ethyl-N '-(3-dimethylaminopropyl) carbodiimide, N, N'-carbonylbis (2-methylimidazole), pentamethyleneketene-N-cyclohexylimine, diphenylketene- N-cyclohexylimine, ethoxyacetylene, 1-alkoxy-1-chloroethylene, trimethyl phosphite, ethyl polyphosphate, isopropyl polyphosphate, phosphorus oxychloride, diphenylphosphoryl azide, thionyl chloride, oxalyl chloride, lower alkyl haloformate (for example, , Ethyl chloroformate, isopropyl chloroformate, etc.), triphenylphosphine, N-hydroxybenzotriazole, 1- (p-chlorobenzenesulfonyloxy) -6-chloro-1H-benzotriazole, N, N-di Chill formamide with thionyl chloride, phosgene, trichloromethyl chloroformate, called Birusumaiya was prepared by the reaction of such phosphorus oxychloride - to carry out the reaction in the presence of a conventional condensing agent such as reagents desired. The reaction is also performed with alkali metal bicarbonate, tri C _1-6 alkylamine, pyridine, N—C _1-6
It may be carried out in the presence of an inorganic or organic base such as alkylmorpholine, N, N-diC _1-6 alkylbenzylamine and the like. The reaction temperature is not particularly limited, but the reaction is usually performed under cooling, at room temperature or under heating. The structural formulas of the compounds obtained by the examples described below are shown below.

[0072]

[Table 1]

[0073]

[Table 2]

[0074]

[Table 3]

As the peptide aldehyde compound having a strong cysteine protease inhibitory activity, a novel compound represented by the following general formula (VI) can also be used.

[0076]

Embedded image

(Wherein each symbol has the same meaning as described above) When the amino acid used in the present invention has an optical isomer, the L-form is indicated unless otherwise specified.

The aryl group having 6 to 10 carbon atoms for R ¹¹ includes, for example, phenyl, naphthyl, pentaphenyl,
Indenyl, azulenyl and the like can be mentioned. Preferably, they are phenyl and naphthyl. Examples of the substituent which the aryl group may have include a halogen atom (such as fluorine and chlorine), alkyl having 1 to 5 carbons, trifluoromethyl, alkoxy having 1 to 5 carbons, hydroxyl, and 2 to 5 carbons. Acyloxy, carboxyl and carbon number 2-5
Acyl group. Preferred are a halogen atom and an alkyl group having 1 to 5 carbon atoms. More preferably,
Fluorine, chlorine and methyl. As preferable specific examples of R ¹¹ is 4-fluorophenyl, 4-chlorophenyl, p- tolyl and 2-naphthyl.

The alkyl group having 1 to 4 carbon atoms represented by R ¹² or R ¹³ includes, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
c-butyl, tert-butyl and the like. Preferred are propyl, isopropyl and tert-butyl. More preferably, it is isopropyl. R ¹² and R ¹³ are
Preferably one of R ¹² or R ¹³ is hydrogen and the other is propyl, isopropyl, isobutyl or tert-butyl, more preferably R ¹² is propyl, isopropyl,
Isobutyl or tert-butyl, wherein R ¹³ is hydrogen,
More preferably, R ¹² is isopropyl and R ¹³ is hydrogen. R ¹² and R ¹³ may be linked to each other to form 3 to 7 carbon atoms
Examples of the ring include cyclopropylidene, cyclobutylidene, cyclopentylidene, cyclohexylidene, cycloheptylidene and the like. Particularly, cyclohexylidene is preferable.

The lower alkyl group represented by R ¹⁴ includes
Linear, branched or cyclic ones having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, Hexyl, 4-methylpentyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 2-ethylbutyl and the like. Preferred are methyl and isobutyl.

The aryl group which may be substituted with the lower alkyl group includes, for example, phenyl, 1-naphthyl, 2
-Naphthyl and the like. Particularly, phenyl is preferable. Examples of the cycloalkyl group which may be substituted with the lower alkyl group include cyclopropane, cyclobutane, cyclopentane and cyclohexane. Particularly, cyclohexane is preferable. Further, examples of the aromatic heterocyclic residue which may be substituted with the lower alkyl group include a monocyclic heterocyclic residue and a condensed heterocyclic residue substituted with oxygen, nitrogen and sulfur atoms. Examples of the monocyclic heterocyclic residue include, for example, pyrrolyl, furanyl, thiophenyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, and pyridyl. Indazolyl, quinazolinyl, phthalazinyl, quinoxalinyl and the like can be mentioned. In particular, indolyl is preferable.

Preferred specific examples of R ¹⁴ include:
Isobutyl, benzyl, cyclohexylmethyl, indol-3-ylmethyl.

The salt of the compound represented by the general formula (VI) is preferably a physiologically acceptable salt.
Examples thereof include salts with inorganic bases, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. Preferred examples of the salt with an inorganic base include, for example, alkali metal salts such as sodium salt and potassium salt;
Alkaline earth metal salts such as calcium salts and magnesium salts; and aluminum salts and ammonium salts. Preferred examples of the salt with an organic base include salts with trimethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, N, N-dibenzylethylenediamine, and the like. Preferable examples of salts with inorganic acids include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Preferred examples of salts with organic acids include, for example, formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfone And salts with acids, p-toluenesulfonic acid and the like. Preferred examples of the salt with a basic amino acid include, for example, salts with arginine, lysine, ornithine and the like. Preferred examples of the salt with an acidic amino acid include, for example, salts with aspartic acid, glutamic acid, and the like. Is mentioned. Compound (VI) is prepared, for example, by the following reaction formula

[0084]

Embedded image

(Wherein each symbol is as defined above).

Examples of the sulfonyl chloride represented by the general formula (VII) [hereinafter sometimes referred to as compound (VII)] include, for example, naphthalenesulfonyl chloride, toluenesulfonyl chloride, fluorobenzenesulfonyl chloride, chlorobenzenesulfonyl chloride, Bromobenzenesulfonyl chloride, benzenesulfonyl chloride and the like can be mentioned.

As the general formula (VIII) [hereinafter sometimes referred to as compound (VIII)], for example, glycine,
Alanine, valine, D-valine, norvaline, leucine, isoleucine, norleucine, tert-leucine, 1
-Aminocyclopropanecarboxylic acid, 1-aminocyclobutanecarboxylic acid, 1-aminocyclopentanecarboxylic acid, 1-aminocyclohexanecarboxylic acid and the like. The reaction between compound (VII) and compound (VIII) can be carried out by a generally known method, for example, Schotten-Baumann (Shot
ten-Baumann) reaction.

The compound represented by formula (IX) and N-hydroxysuccinimide are dissolved in a commonly used organic solvent (eg, tetrahydrofuran, dichloromethane, chloroform, ethyl acetate, etc.) and condensed with a condensing agent. . As the condensing agent, for example, N, N-dicyclohexylcarbodiimide and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride are preferably used.

An amino alcohol represented by the general formula (XI) [hereinafter sometimes referred to as compound (XI). ] Include, for example, vallinol, leucinol, D-leucinol, phenylalaninol, tryptophanol,
(S) -2-amino-3-cyclohexyl-1-propanol and the like.

The compound represented by the general formula (X) and the compound (XI) are dissolved in a solvent such as tetrahydrofuran, dichloromethane, chloroform and ethyl acetate.
The reaction is performed in the presence of a base (triethylamine, pyridine, etc.).

Further, when the compound represented by the general formula (XII) is oxidized with an oxidizing agent (a sulfur trioxide pyridine complex, oxalyl chloride, chromic acid-pyridine, etc.), a novel compound (VI) can be produced. .

The reaction temperature is not particularly limited.
It is performed under cooling, at room temperature or under heating. The structural formulas of the compounds obtained by the examples described below are shown below.

[0093]

[Table 4]

[0094]

[Table 5]

[0095]

[Table 6]

The cysteine protease inhibiting compounds can be administered systemically or locally. In addition to oral administration systemically, parenteral administration such as intravenous injection, subcutaneous injection, and intramuscular injection can be used. It can be administered topically to the skin, mucous membranes, intranasally, intraocularly, and the like.

The cysteine protease inhibiting compounds can be formulated for use in pharmaceutical compositions. Compositions for oral administration to humans include, for example, powders, granules, tablets, capsules, syrups, solutions and the like. When the composition is formulated as a powder, granules, tablets or the like, any pharmaceutical carrier suitable for formulating a solid composition,
For example, excipients (starch, glucose, fructose, sucrose, etc.), lubricants (magnesium stearate, etc.), disintegrants (starch,
Crystalline cellulose, etc.), binders (starch, gum arabic, etc.)
And the like, and may be coated with a coating agent (gelatin, sucrose, etc.). When the composition is formulated as a syrup or a liquid, for example, a stabilizer (eg, sodium edetate), a suspending agent (eg, gum arabic, carmellose), a flavoring agent (eg, simple syrup, glucose), and a fragrance can be used. It can be appropriately selected and used. Parenterally formulated compositions include injections, suppositories and the like. When the composition is formulated as an injection, for example, a solvent (such as distilled water for injection), a stabilizer (such as sodium edetate), an isotonic agent (such as sodium chloride, glycerin, and mannitol), and a pH adjuster (such as hydrochloric acid) , Citrate, sodium hydroxide, etc.) and suspending agents (eg, methylcellulose). When formulated as a suppository, for example, a suppository base (eg, cocoa butter, macrogol, etc.) may be appropriately selected. Can be used. Examples of the composition for external use include ointments, creams, lotions, nasal drops, eye drops and the like. In these external compositions, in addition to the inhibitory compound, for example, an ointment base (Vaseline, lanolin, etc.), a solvent (physiological saline, purified water, etc.), a stabilizer (Sodium edetate, citric acid, etc.), a wetting agent ( Glycerin, etc.), emulsifier (polyvinylpyrrolidone, etc.), suspending agent (hydroxypropylmethylcellulose, methylcellulose, etc.), surfactant (polysorbate 80, polyoxyethylene hydrogenated castor oil, etc.), preservative (benzalkonium chloride, parabens) , Chlorobutanol, etc.), buffers (boric acid, borax, sodium acetate, citrate buffer, phosphate buffer, etc.), tonicity agents (sodium chloride, glycerin, mannitol, etc.), pH adjusters (hydrochloric acid, A known compound such as sodium hydroxide can be appropriately selected and used.

The angiogenesis inhibitor of the present invention may be formulated with other pharmaceutical ingredients, such as anti-inflammatory drugs, anti-tumor drugs, antibacterial drugs and the like.

The dose of the cysteine protease inhibitor compound varies depending on the target disease, symptom, subject of administration, administration method and the like, but the dose per dose is usually 1 to 500 mg, preferably 10 to 200 mg for oral administration. For injections, usually 0.1 to 100 mg, preferably 1 to 50 mg is used for treating diseases. Also, when used locally,
Usually 0.001 to 1.0 w / v%, preferably 0.01
Ophthalmic solution adjusted to 0.5% w / v% once at a time of 20 to 50%
μ1, it is preferable to instill 5 to 6 times a day.

The present invention will be described in more detail with reference to the following Examples and Test Examples, but the present invention is not limited thereto.

Example 1 (Tablets) E-64 30 mg Lactose 80 mg Starch 17 mg Magnesium stearate 3 mg A tablet is formed by a conventional method using the above material for one tablet.
Sugar coating may be applied as necessary.

Example 2 (Injection) Leupeptin 100 mg Sodium chloride 900 mg 1N sodium hydroxide Appropriate amount Distilled water for injection Total 100 ml The above components are mixed by a conventional method to prepare an injection.

Example 3 (Eye drops) 27-mer calpastatin peptide 1 g Boric acid 0.7 g Borax qs Sodium chloride 0.5 g Hydroxypropyl methylcellulose 0.1 g Sodium EDTA 0.02 g Benzalkonium chloride 0.0 g 005 g Sterile purified water A total of 100 ml or more components are mixed by a conventional method to prepare an ophthalmic suspension.

Reference Example 1 N-tert-butoxycarbonylphenylalanine (53
g, 0.2 mol) and p-nitrophenol (27.8 g, 0.2 mo
l) in ethyl acetate (200 ml) solution under ice-cooling, N, N'-
A solution of dicyclohexylcarbodiimide (41.2 g, 0.2 mol) in ethyl acetate (100 ml) was added dropwise, and the mixture was stirred as it was for 3 hours, and further stirred at room temperature for 20 hours. N deposited,
N'-Dicyclohexylcarbodiurea was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was recrystallized from ethyl acetate-hexane to give N-tert-butoxycarbonylphenylalanine p-nitrophenyl ester [61.7 g, 80% (Hereinafter, unless otherwise specified, the values are shown in% by weight)].

Reference Example 2 N-tert-butoxycarbonylleucine (6.94 g, 30 m
mol) and N-hydroxysuccinimide (3.45 g, 30 mmo
l) in dioxane (50 ml) solution under ice-cooling with N-ethyl-
A solution of N ′-(3-dimethylaminopropyl) carbodiimide hydrochloride (5.75 g, 30 mmol) in dioxane was added dropwise,
The mixture was stirred for 20 minutes as it was, and further stirred at room temperature for 24 hours. The reaction solution was poured into cold water, extracted with ethyl acetate,
The organic layer washed with a w / v% citric acid aqueous solution, a 10 w / v% sodium hydrogencarbonate aqueous solution and a saturated saline solution in that order was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and the residue was recrystallized from isopropyl ether. -Tert-Butoxycarbonylleucine N-hydroxysuccinimide ester (7.77 g, 78.9%) was obtained.

Reference Example 3 1- (4-Fluorophenyl) piperazine dihydrochloride (2.
53 g, 10 mmol) of N, N'-dimethylformamide (40
ml) solution, triethylamine (2.8 ml, 20 mmol) and N
-Tert-Butoxycarbonylphenylalanine p-nitrophenyl ester (2.65 g, 10 mmol) was sequentially added, and the mixture was stirred at room temperature overnight. The reaction solution was poured into cold water, extracted with ethyl acetate, and the organic layer washed with a 1% (w / v) aqueous ammonia solution, a saturated saline solution, 0.1N hydrochloric acid, a saturated saline solution, a saturated sodium hydrogen carbonate aqueous solution, and a saturated saline solution in that order was dried. After drying over magnesium sulfate, the mixture was concentrated under reduced pressure, and the residue was subjected to silica gel chromatography. Elution was performed with chloroform: methanol (50: 1, volume ratio, the same applies hereinafter), and 2- (4
-(4-fluorophenyl) -1-piperazinyl) -2
-Oxo-1- (phenylmethyl) ethylcarbamic acid 1,1-dimethylethyl ester (2.7 g, 92.2%) was obtained as a colorless oil.

Reference Example 4 In the same manner as in Reference Example 3 except that 1- (o-fluorophenyl) piperazine monohydrochloride was used instead of 1- (4-fluorophenyl) piperazine dihydrochloride, 2- ( 4-
(2-fluorophenyl) -1-piperazinyl) -2-
Oxo-1- (phenylmethyl) ethylcarbamic acid
1,1-dimethylethyl ester (1.89 g, 88.4%) was obtained.

Reference Example 5 1- (4-fluorophenyl) piperazine dihydrochloride (0.1%)
91 g, 3 mmol) and N-tert-butoxycarbonylleucine N-hydroxysuccinimide ester (0.99 g,
Triethylamine (1.3 ml, 9 mmol) was added to a mixture of 3 mmol) of dichloromethane (50 ml), and the mixture was stirred at room temperature for 20 hours. The reaction solution was washed successively with 0.1N hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate, water and saturated saline, and the organic layer was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and the residue was subjected to silica gel chromatography. Elution with ethyl acetate: hexane (1: 1) gave 2- (4- (4-fluorophenyl) -1-piperazinyl) -2-oxo-1-.
(2-methylpropyl) ethylcarbamic acid 1,1-
Dimethyl ethyl ester (1.05 g, 89.0%) was obtained as a colorless oil.

Reference Example 6 The same operation as in Reference Example 5 was carried out except that 4-phenylpiperazine was used instead of 1- (4-fluorophenyl) piperazine dihydrochloride, to give 2- (4-phenyl-1-piperazinyl). -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester (7.99
g, 99%).

Reference Example 7 In the same manner as in Reference Example 5 except that 1-dimethylsulfamoylpiperazine was used instead of 1- (4-fluorophenyl) piperazine dihydrochloride, 2- (4-dimethylsulfuryl) Moyl-1-piperazinyl) -2-oxo-1-
(2-methylpropyl) ethylcarbamic acid 1,1-
Dimethyl ethyl ester (7.19 g, 88.4%) was obtained.

Reference Example 8 The same operation as in Reference Example 5 was carried out except that p-toluenesulfonylpiperazine was used instead of 1- (4-fluorophenyl) piperazine dihydrochloride, to obtain 2- (4- (4-methylphenyl) piperazine. Sulfonyl) -1-piperazinyl) -2-oxo-1
-(2-methylpropyl) ethylcarbamic acid 1,1
-Dimethylethyl ester (6.95 g, 79.4%) was obtained.

Reference Example 9 In the same manner as in Reference Example 5 except that 1- (2-chlorophenyl) piperazine was used instead of 1- (4-fluorophenyl) piperazine dihydrochloride, 2- (4- (2 -Chlorophenyl) -1-piperazinyl) -2-oxo-1-
(2-methylpropyl) ethylcarbamic acid 1,1-
Dimethyl ethyl ester (5.70 g, 95.5%) was obtained.

Reference Example 10 In place of 1- (4-fluorophenyl) piperazine dihydrochloride, 1- (m-chlorophenyl) piperazine monohydrochloride was used, and the same operation as in Reference Example 5 was carried out to obtain 2- (4 -(3-
(Chlorophenyl) -1-piperazinyl) -2-oxo-
1- (2-methylpropyl) ethylcarbamic acid 1,
1-dimethylethyl ester (2.63 g, 88.4%) was obtained.

Reference Example 11 The same operation as in Reference Example 5 was carried out except that 1- (4-chlorophenyl) piperazine monohydrochloride was used instead of 1- (4-fluorophenyl) piperazine dihydrochloride, to obtain 2- (4 − (4
-Chlorophenyl) -1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid
1,1-Dimethylethyl ester (2.83 g, 94.8%) was obtained.

Reference Example 12 N- (p-methoxyphenyl) piperazine succinate was used in place of 1- (4-fluorophenyl) piperazine dihydrochloride, and the same operation as in Reference Example 5 was carried out to give 2- (p-methoxyphenyl) piperazine succinate. 4-
(4-methoxyphenyl) -1-piperazinyl) -2-
Oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester (2.73 g, 92.3
%).

Reference Example 13 2- (4- (4-Fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid 1,1-dimethylethyl ester (2.7 g,
To a solution of 6.3 mmol) in ethyl acetate (20 mmol) was added dropwise a solution of 4N hydrogen chloride in ethyl acetate (20 ml) under ice cooling, and the mixture was stirred at room temperature overnight. The precipitated crystals were collected by filtration and recrystallized from ethanol + diethyl ether to give 1- (2-amino-1-
Oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride (2.2 g, 96.1%) was obtained as pale yellow crystals.

Reference Example 14 2- (4- (4-fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Using 4- (2-fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid 1,1-dimethylethyl ester, the same operation as in Reference Example 13 was performed to obtain 1- ( 2-amino-
1-oxo-3-phenylpropyl) -4- (2-fluorophenyl) piperazine hydrochloride (1.3 g, 99.1%) was obtained as white crystals.

Reference Example 15 2- (4- (4-fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Using 4- (2-fluorophenyl) -1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester, the same operation as in Reference Example 13 was performed to obtain 1 -(2-Amino-4-methyl-1-oxopentyl) -4- (4-fluorophenyl) piperazine hydrochloride (0.56 g, 70.4%) was obtained as white crystals.

Reference Example 16 2- (4- (4-Fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Reference Example 13 using 4-phenyl-1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester
The same operation as described above was carried out, and 1- (2-amino-4-methyl-1
-Oxopentyl) -4-phenylpiperazine hydrochloride (6.5 g, 99.2%) was obtained as white crystals.

Reference Example 17 2- (4- (4-fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Using 4-dimethylsulfamoyl-1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester, the same operation as in Reference Example 13 was performed to obtain 1- ( 2-amino-
4-Methyl-1-oxopentyl) -4-dimethylsulfamoylpiperazine hydrochloride (5.0 g, 83.3%) was obtained as white crystals.

Reference Example 18 2- (4- (4-fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( 4- (4-methylphenylsulfonyl) -1
-Piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester was used, and the same operation as in Reference Example 13 was performed to obtain 1- (2
-Amino-4-methyl-1-oxopentyl) -4-
(4-Methylphenylsulfonyl) piperazine hydrochloride (4.83 g, 78.4%) was obtained as white crystals.

Reference Example 19 2- (4- (4-Fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Using 4- (2-chlorophenyl) -1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester, the same operation as in Reference Example 13 was performed to give 1- (2-amino-
4-Methyl-1-oxopentyl) -4- (2-chlorophenyl) piperazine hydrochloride (1.54 g, 62.6%) was obtained as white crystals.

Reference Example 20 2- (4- (4-fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Using 4- (3-chlorophenyl) -1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester, the same operation as in Reference Example 13 was performed to give 1- (2-amino-
4-Methyl-1-oxopentyl) -4- (3-chlorophenyl) piperazine hydrochloride (1.40 g, 65.7%) was obtained as white crystals.

Reference Example 21 2- (4- (4-Fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Using 4- (4-chlorophenyl) -1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester, the same operation as in Reference Example 13 was performed to give 1- (2-amino-
4-Methyl-1-oxopentyl) -4- (4-chlorophenyl) piperazine hydrochloride (1.50 g, 65.3%) was obtained as white crystals.

Reference Example 22 2- (4- (4-Fluorophenyl) -1-piperazinyl) -2-oxo-1- (phenylmethyl) ethylcarbamic acid Instead of 1,1-dimethylethyl ester, 2- ( Using 4- (4-methoxyphenyl) -1-piperazinyl) -2-oxo-1- (2-methylpropyl) ethylcarbamic acid 1,1-dimethylethyl ester, the same operation as in Reference Example 13 was carried out to give 1 -(2-Amino-4-methyl-1-oxopentyl) -4- (4-methoxyphenyl) piperazine hydrochloride (2.21 g, 87.4%) was obtained as white crystals.

Reference Example 23 N-tert-butoxycarbonyl-L-valine (2.27 g,
10 mmol) and 1- (2-chlorophenyl) piperazine (2.
00 g, 10 mmol) of N, N-dimethylformamide (50 m
l) 1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (2.2 g, 11 mmol
l) and 1-hydroxybenzotriazole (1.5 g, 11
mmol) in dichloromethane (50 ml) was added dropwise. After the reaction mixture was stirred at room temperature for 15 hours, dichloromethane was distilled off under reduced pressure, and ethyl acetate (200 ml) was added to the residue. The ethyl acetate layer was washed sequentially with a 10% w / v aqueous solution of citric acid, a saturated aqueous solution of sodium bicarbonate, and a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate.
The residue was subjected to silica gel chromatography, eluting with ethyl acetate: n-hexane (1: 2) to give 2- (4-
(2-chlorophenyl) piperazinyl) -2-oxo-
(s) -1- (2-propyl) ethylcarbamic acid 1,1
-Dimethyl ethyl ester was obtained. This colorless oil was dissolved in ethyl acetate (50 ml), and a 4N solution of hydrogen chloride in ethyl acetate (50 ml) was added dropwise under ice-cooling, followed by stirring at room temperature for 3 hours. The product was filtered, washed with ethyl acetate-n-hexane (1: 1) and 1-((s) -2-amino-3-methyl-1-oxobutyl) -4- (2-chlorophenyl) piperazine hydrochloride. The salt (3.32 g, 95.8%) was obtained as colorless crystals.

Reference Example 24 Using the same method as in Reference Example 23, 1- (2-amino-1-oxoethyl) -4- (2-chlorophenyl) piperazine hydrochloride was prepared from N-tert-butoxycarbonyl-glycine. 2.4 g, 90.4%) as colorless crystals.

Reference Example 25 Using the same method as in Reference Example 23, N-tert-butoxycarbonyl-L-alanine was used to give 1-((s) -2-amino-1-oxopropyl) -4- (2 -Chlorophenyl) piperazine hydrochloride (1.7 g, 58.8%) was obtained as colorless crystals.

Reference Example 26 Using the same method as in Reference Example 23, N-tert-butoxycarbonyl-L-isoleucine was used to give 1-((s) -2-
Amino-3-methyl-1-oxopentyl) -4- (2
-Chlorophenyl) piperazine hydrochloride (3.4 g, 90.2
%) As colorless crystals.

Reference Example 27 Using the same method as in Reference Example 23, N-tert-butoxycarbonyl-β-alanine was converted to 1- (3-amino-1-amino).
Oxopropyl) -4- (2-chlorophenyl) piperazine hydrochloride (2.9 g, 90.0%) was obtained as colorless crystals.

Reference Example 28 Using the same method as in Reference Example 23, 1- (2-methylamino-1-oxoethyl) -4- (2-chlorophenyl) piperazine hydrochloride was obtained from N-tert-butoxycarbonyl-sarcosine. (3.0 g, 93.3%) was obtained as colorless crystals.

Reference Example 29 Using the same method as in Reference Example 23, N-tert-butoxycarbonyl-L-proline was used to give 1- (1- (2-pyrrolidinyl) -1-oxomethyl) -4- (2- Chlorophenyl) piperazine hydrochloride (4.3. G, 98.0%) was obtained as colorless crystals.

Reference Example 30 Using the same method as in Reference Example 23, N-tert-butoxycarbonyl- (s) -acetamidomethyl) -L-cysteine was used to give 1-((s) -2-amino-3- (Acetylaminomethylthio) -1-oxopropyl) -4- (2-chlorophenyl) piperazine hydrochloride (4.0 g, 95.9%) was obtained as colorless crystals.

Reference Example 31 Using the same method as in Reference Example 23, N-tert-butoxycarbonyl-L-methionine was used to give 1-((s) -2-amino-4-methylthio-1-oxobutyl) -4. -(2
-Chlorophenyl) piperazine hydrochloride (3.7 g, 97.1
%) As colorless crystals.

Reference Example 32 Using a method similar to that of Reference Example 23, N-tert-butoxycarbonyl-L-glutamine was used to give 1-((s) -2-amino-4-carbamoyl-1-oxobutyl) -4. −
(2-Chlorophenyl) piperazine hydrochloride (2.7 g, 60.
7%) as colorless crystals.

Example 4 N, N-dimethylformamide (1.82 g, 5 mmol) of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride (20%) ml) solution in solution with triethylamine (0.697 ml, 5 mmol)
And stirred at room temperature for 10 minutes, followed by a method such as TAMAI [Ch
em. Pharm. Bull., 35, 1098 (1987)], and added p-nitrophenyl L-trans-epoxysuccinic acid ethyl ester (1.41 g, 5 mmol) at room temperature.
Stirred for hours. The reaction solution was poured into cold water, extracted with ethyl acetate, 1 w / v% aqueous ammonia, saturated saline,
0.1N hydrochloric acid, saturated saline, saturated sodium hydrogen carbonate,
The organic layer washed with saturated saline in this order was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and the residue was subjected to silica gel chromatography. Ethyl acetate: hexane (1:
1), and eluted with (2s, 3s) -3-[[[[(1s)
-1-[[4- (4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.2 g, 51.1%; compound 1) was obtained.

¹ H NMR (CDCl ₃ ) δ: 1.31 (t, 3H, J = 7.0 H
_{z, -C-CH 3),} 2.42~2.50 (m, 1H, piperazine ring), 2.
83 〜2.94 (m, 2H, piperazine ring), 3.00 (d, 2H, J =
7.6 Hz, ph-CH ₂ -C-), 2.97 ~ 3.07 (m, 1H, piperazine r
ing), 3.14-3.22 (m, 1H, piperazine ring), 3.35
(d, 1H, J = 1.9 Hz, epoxy ring), 3.43-3.52 (m, 1H, p
iperazine ring), 3.64 (d, 1H, J = 1.9 Hz, epoxy rin
g), 3.71 (t, 2H, J = 5.4 Hz, piperazine ring), 4.25
(dq, 2H, J = 7.3, 2.6 Hz, -O-CH ₂ -C), 5.18 (q, 1H, J =
5.3 Hz, -N-CH-CO), 6.75 to 6.83 (m, 2H, aromatic),
6.91 to 7.04 (m, 2H, aromatic, 1H, NH), 7.17 to 7.34
(m, 5H, aromatic).

Example 5 Instead of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-1-oxo-3 Using (-phenylpropyl) -4- (2-fluorophenyl) piperazine hydrochloride, the same operation as in Example 4 was performed to obtain (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (0.83 g, 58.6%;
Compound 2) was obtained.

¹ H NMR (CDCl ₃ ) δ: 1.31 (t, 3H, J = 7.1 H
_{z, -C-CH 3),} 2.42~2.49 (m, 1H, piperazine ring), 2.
72 〜2.90 (m, 2H, piperazine ring), 3.02 (d, 2H, J =
8.3 Hz, ph-CH ₂ -C-), 2.94-3.10 (m, 1H, piperazine r
ing), 3.17-3.29 (m, 1H, piperazine ring), 3.36
(d, 1H, J = 1.7 Hz, epoxy ring), 3.44-3.57 (m, 1H, p
iperazine ring), 3.65 (d, 1H, J = 1.3 Hz, epoxy rin
g), 3.66 to 3.80 (m, 2H, piperazine ring), 4.25 (dq,
2H, J = 7.1, 2.3 Hz, -O-CH ₂ -C), 5.19 (q, 1H, J = 7.6 H
z, -N-CH-CO), 6.8 (t, 1H, J = 8.3 Hz, -NH-), 6.93-7.1
1 (m, 4H, aromatic), 7.18 to 7.34 (m, 5H, aromatic).

Example 6 Instead of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 Using (-oxopentyl) -4- (4-fluorophenyl) piperazine hydrochloride, the same operation as in Example 4 was performed to obtain (2s,
3s) -3-[[[[(1s) -1-[[4- (4-fluorophenyl) -1-piperazinyl] carbonyl]-
3-Methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (0.30 g, 45.6%; compound 3)
I got

¹ H NMR (CDCl ₃ ) δ: 0.93 (d, 3H, J = 6.3 H
z, -C-CH ₃ ), 1.00 (d, 3H, J = 6.3 Hz, -C-CH ₃ ), 1.32
(t, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.40-1.63 (m, 3H, -CC
H ₂ -CH-C ₂ ), 3.06-3.15 (m, 4H, piperazine ring), 3.4
9 (d, 1H, J = 1.7 Hz, epoxy ring), 3.60-3.89 (m, 4
H, piperazine ring), 3.68 (d, 1H, J = 1.7 Hz, epoxy r
ing), 4.26 (dq, 2H, J = 7.3, 3.3 Hz, -O-CH ₂ -C), 5.03
(dt, 1H, J = 8.91, 4.3 Hz, -N-CH-CO), 6.85 to 7.03
(m, 5H, aromatic and -NH).

Example 7 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 -Oxopentyl) -4-phenylpiperazine hydrochloride,
The same operation as in Example 4 was performed, and (2s, 3s) -3-
[[[[[1s) -1-[(4-phenyl-1-piperazinyl) carbonyl] -3-methyl] butyl] amino]
Carbonyl] oxiranecarboxylic acid ethyl ester (3.
96 g, 63.3%; compound 4) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.93 (d, 3H, J = 6.3 H
z, -C-CH ₃ ), 1.00 (d, 3H, J = 6.3 Hz, -C-CH ₃ ), 1.32
(t, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.40-1.63 (m, 3H, -CC
H ₂ -CH-C ₂ , 3.16 to 3.24 (m, 4H, piperazine ring), 3.4
9 (d, 1H, J = 1.7 Hz, epoxy ring), 3.60-3.89 (m, 4H,
piperazine ring), 3.68 (d, 1H, J = 1.7 Hz, epoxy rin
g), 4.26 (dq, 2H, J = 7.3, 3.3Hz, -O-CH ₂ -C), 5.03 (d
t, 1H, J = 8.9, 4.3Hz, -N-CH-CO), 6.90-6.95 (m, 4
H, aromatic and -NH), 7.25 to 7.33 (m, 2H, aromati
c).

Example 8 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 (Oxopentyl) -4-dimethylsulfamoylpiperazine hydrochloride, the same operation as in Example 4 was carried out to obtain (2s, 3
s) -3-[[[[(1s) -1-[(4-dimethylsulfamoyl-1-piperazinyl) carbonyl] -3-
Methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (3.7 g, 82.6%; compound 5) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.92 (d, 3H, J = 6.3 H
_{z, -C-CH 3),} 0.89 (d, 3H, J = 6.3 Hz, -C-CH 3), 1.32
(t, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.38-1.60 (m, 3H, -CC
H ₂ -CH-C ₂ ), 2.85 (s, 6H, -N-CH ₃ ), 3.15 to 3.38 (m, 4H,
piperazine ring), 3.48 (d, 1H, J = 1.7 Hz, epoxy rin
g), 3.52-3.68 (m, 3H, piperazine ring), 3.67 (d, 1
H, J = 1.7 Hz, epoxy ring), 3.79-3.87 (m, 1H, pipera
zine ring), 4.27 (dq, 2H, J = 7.3, 4.0 Hz, -O-CH _2-
C), 4.96 (dt, 1H, J = 8.9, 4.3 Hz, -N-CH-CO), 6.90
(d, 1H, J = 8.6 Hz, -NH-).

Example 9 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 -Oxopentyl) -4- (4-methylphenylsulfonyl)
Using piperazine hydrochloride, the same operation as in Example 4 was performed,
(2s, 3s) -3-[[[[(2s) -1-[[4-
(4-Methylphenylsulfonyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (4.31
g, 95.1%; compound 6) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.87 (d, 3H, J = 6.3 H
_{z, -C-CH 3),} 0.94 (d, 3H, J = 6.3 Hz, -C-CH 3), 1.30
(t, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.31 to 1.55 (m, 3H, -CC
H ₂ -CH-C ₂ ), 2.45 (s, 6H, -ph-CH ₃ ), 2.70 to 2.84 (m, 2
H, piperazine ring), 3.22 ~ 3.54 (m, 4H, piperazine
ring), 3.43 (d, 1H, J = 1.7 Hz, epoxy ring), 3.61 (d,
1H, J = 2.0 Hz, epoxy ring), 3.68 〜 3.78 (m, 1H, pip
erazine ring), 3.96 to 4.06 (m, 1H, piperazine rin
g), 4.25 (dq, 2H, J = 7.3, 4.0 Hz, -O-CH ₂ -C), 4.87 (d
t, 1H, J = 9.2, 4.0 Hz, -N-CH-CO), 6.81 (d, 1H, J = 8.6
Hz, -NH-), 7.35 (d, 2H, J = 7.9 Hz, aromatic), 7.63
(d, 2H, J = 8.3 Hz, aromatic).

Example 10 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 (Oxopentyl) -4- (2-chlorophenyl) piperazine hydrochloride and the same operation as in Example 4 to give (2s, 3
s) -3-[[[[(1s) -1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] -3-
Methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (0.65 g, 35.9%; compound 7) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.93 (d, 3H, J = 6.3 H
z, -C-CH ₃ ), 1.01 (d, 3H, J = 6.3 Hz, -C-CH ₃ ), 1.32
(t, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.41 to 1.61 (m, 3H, -CC
H ₂ -CH-C ₂ ), 2.96--3.10 (m, 4H, piperazine ring), 3.4
9 (d, 1H, J = 2.0 Hz, epoxy ring), 3.61 to 3.81 (m, 3
H, piperazine ring), 3.68 (d, 1H, J = 2.0 Hz, epoxy r
ing), 3.90 to 3.98 (m, 1H, piperazine ring), 4.27 (d
q, 2H, J = 7.3, 4.0 Hz, -O-CH ₂ -C), 5.03 (dt, 1H, J =
8.9,4.3 Hz, -N-CH-CO), 6.91 (d, 1H, J = 8.6 Hz, -NH
-), 7.00 to 7.06 (m, 2H, aromatic), 7.21 to 7.27 (m,
1H, aromatic), 7.37 to 7.41 (m, 1H, aromatic).

Example 11 Instead of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 (Oxopentyl) -4- (3-chlorophenyl) piperazine hydrochloride and the same operation as in Example 4 to give (2s, 3
s) -3-[[[[(1s) -1-[[4- (3-chlorophenyl) -1-piperazinyl] carbonyl] -3-
Methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.24 g, 68.4%; compound 8) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.93 (d, 3H, J = 6.3 H
z, -C-CH ₃ ), 1.00 (d, 3H, J = 6.3 Hz, -C-CH ₃ ), 1.32
(t, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.42 to 1.63 (m, 3H, -CC
H ₂ -CH-C ₂ ), 3.17 ~ 3.25 (m, 4H, piperazine ring), 3.4
8 (d, 1H, J = 2.0 Hz, epoxy ring), 3.60-3.90 (m, 4
H, piperazine ring), 3.67 (d, 1H, J = 2.0 Hz, epoxy r
ing), 4.27 (dq, 2H, J = 7.3, 4.0 Hz, -O-CH ₂ -C), 5.02
(dt, 1H, J = 8.9, 4.3 Hz, -N-CH-CO), 6.77 ~ 6.81 (m, 1
H, aromatic), 6.86-6.89 (m, 3H, aromatic and -N
H), 7.16-7.22 (m, 1H, aromatic).

Example 12 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 Using (oxopentyl) -4- (4-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was carried out to obtain (2s, 3
s) -3-[[[[(1s) -1-[[4- (4-chlorophenyl) -1-piperazinyl] carbonyl] -3-
Methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (0.55 g, 27.1%; compound 9) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.93 (d, 3H, J = 6.3 H
z, -C-CH ₃ ), 1.0 (d, 3H, J = 6.3 Hz, -C-CH ₃ ), 1.32 (t,
3H, J = 7.3 Hz, -C-CH ₃ ), 1.42 to 1.63 (m, 3H, -C-CH _2-
CH-C ₂ ), 3.12 ~ 3.20 (m, 4H, piperazine ring), 3.48
(d, 1H, J = 2.0 Hz, epoxy ring), 3.60-3.90 (m, 4H,
piperazine ring), 3.67 (d, 1H, J = 2.0 Hz, epoxy rin
g), 4.27 (dq, 2H, J = 7.3, 4.0 Hz, -O-CH ₂ -C), 5.02 (d
t, 1H, J = 8.9, 4.3 Hz, -N-CH-CO), 6.83-6.87 (m, 2H,
aromatic), 6.90 (d, 1H, J = 9.9 Hz, -NH), 7.21-7.3
(m, 2H, aromatic).

Example 13 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-4-methyl-1 (Oxopentyl) -4- (4-methoxyphenyl) piperazine hydrochloride and the same operation as in Example 4 to give (2s,
3s) -3-[[[[(1s) -1-[[4- (4-methoxyphenyl) -1-piperazinyl] carbonyl]-
3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (0.94 g, 65.2%; compound 1
0) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.92 (d, 3H, J = 6.6 H
z, -C-CH ₃ ), 1.00 (d, 3H, J = 6.3 Hz, -C-CH ₃ ), 1.32
(t, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.40-1.60 (m, 3H, -CC
H ₂ -CH-C ₂ ), 3.03-3.11 (m, 4H, piperazine ring), 3.4
9 (d, 1H, J = 2.0 Hz, epoxy ring), 3.60-3.88 (m, 4
H, piperazine ring), 3.67 (d, 1H, J = 2.0 Hz, epoxy r
ing), 3.78 (s, 3H, -O-CH ₃ ), 4.27 (dq, 2H, J = 7.3, 4.
0 Hz, -O-CH ₂ -C), 5.03 (dt, 1H, J = 8.9, 4.3 Hz, -N-CH
-CO), 6.83 to 6.96 (m, 5H, aromatic and -NH).

Example 14 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1-((s) -2-amino-3 -Methyl-
Using 1-oxobutyl) -4- (2-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was performed to obtain (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
-2-Methyl] propyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.3 g, 57.4%) was obtained as a colorless oil.

Example 15 Instead of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-amino-1-oxoethyl)-
Using 4- (2-chlorophenyl) piperazine hydrochloride,
The same operation as in Example 4 was performed, and (2s, 3s) -3-
[[[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] methyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.75 g, 53.5%) was obtained as a colorless oil.

Example 16 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1-((s) -2-amino-1 Using (-oxopropyl) -4- (2-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was performed to obtain (2s, 3s)
-3-[[[(1s) -1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.23 g, 55.0%) as a colorless oil As obtained.

Example 17 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1-((s) -2-amino-3 -Methyl-
Using 1-oxopentyl) -4- (2-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was performed,
(2s, 3s) -3-[[[[(1s) -1-[[4-
(2-Chlorophenyl) -1-piperazinyl] carbonyl] -2-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.48 g, 56.6%) was obtained as a colorless oil.

Example 18 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (3-amino-1-oxopropyl)
Using (4- (2-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was carried out to obtain (2s, 3s) -3-
[[[2-[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] ethyl] amino] carbonyl]
Oxirane carboxylic acid ethyl ester (1.16, 52.9%)
Was obtained as a colorless oil.

Example 19 Instead of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (2-methylamino-1-oxoethyl) Using (4- (2-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was performed to obtain (2s, 3s) -3.
-[[[[N-[[4- (2-chlorophenyl) -1-
Piperazinyl] carbonyl] methyl] -N-methyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.55 g, 76.7%) was obtained as a colorless oil.

Example 20 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1- (1- (2-pyrrolidinyl) -1 Using (-oxomethyl) -4- (2-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was carried out to obtain (2s, 3
s) -3-[[(2s) -2-[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] -1-pyrrolidinyl] carbonyl] oxiranecarboxylic acid ethyl ester (1.42 g, 49.9%) was colorless. Obtained as an oil.

Example 21 1-((s) -2-amino-3 was used instead of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride. -(Acetylaminomethylthio) -1-oxopropyl) -4-
Using (2-chlorophenyl) piperazine hydrochloride, the same operation as in Example 4 was performed to obtain (2s, 3s) -3-
[[[[[(1s) -1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] -2-acetylaminomethylthio] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester (1.19 g, 43.7% ) Was obtained as a colorless oil.

Example 22 In place of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride, 1-((s) -2-amino-4 -Methylthio-1-oxobutyl) -4- (2-chlorophenyl)
Using piperazine hydrochloride, the same operation as in Example 4 was performed,
(2s, 3s) -3-[[[[(1s) -1-[[4-
(2-Chlorophenyl) -1-piperazinyl] carbonyl] -4-methylthio] butyl] amino] carbonyl]
Oxirane carboxylic acid ethyl ester (1.54 g, 61.5
%) As a colorless oil.

Example 23 1-((s) -2-amino-4 was used instead of 1- (2-amino-1-oxo-3-phenylpropyl) -4- (4-fluorophenyl) piperazine hydrochloride. -Carbamoyl-1-oxobutyl) -4- (2-chlorophenyl) piperazine hydrochloride was operated in the same manner as in Example 4 to obtain (2s, 3s) -3-[[[[(1s) -1-
[[4- (2-chlorophenyl) -1-piperazinyl]
Carbonyl] -3-carbamoyl] propyl] amino]
Carbonyl] oxiranecarboxylic acid ethyl ester (0.
2 g, 5.8%) as a colorless oil.

Example 24 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Oxirane carboxylic acid ethyl ester (0.5 g, 1.06 mm
ol) in ethanol (20 ml) was added under ice-cooling a 0.1 N sodium hydroxide in ethanol (16 ml), and the mixture was stirred at room temperature for 20 hours. The reaction solution was poured into cold water, acidified with 1N hydrochloric acid, and the precipitated white matter was collected by filtration, dried, and recrystallized from ethyl acetate + hexane to give (2s, 3s) -3.
-[[[[(1s) -1-[[4- (4-fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl] oxiranecarboxylic acid (0.36 g, 77.8%; compound 11) was obtained.

¹ H NMR (CDCl ₃ ) δ: 2.34 to 2.41 (m, 1H, p
iperazine ring), 2.82 to 2.96 (m, 2H, piperazine rin
g), 2.99 〜 3.08 (m, 1H, piperazine ring), 3.06 (d,
2H, J = 7.3 Hz, ph-CH ₂ -C-), 3.16-3.24 (m, 1H, pipera
zine ring), 3.49-3.58 (m, 1H, piperazine ring),
3.55 (d, 1H, J = 1.7 Hz, epoxy ring), 3.57 (d, 1H, J =
1.7 Hz, epoxy ring), 3.71 (t, 2H, J = 5.1 Hz, pipera
zine ring), 4.5-6.0 (brd, 1H, -COOH), 5.23 (q, 1
H, J = 7.9 Hz, -N-CH-CO), 6.74 to 6.82 (m, 2H, aromati
c), 6.91 to 7.00 (m, 2H, aromatic), 7.20 to 7.35 (m, 5
H, aromatic), 8.23 (d, 1H, J = 8.6 Hz, -NH-).

Example 25 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
Example 24 using -fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester.
And the same operation as ((2s, 3s) -3-
[[[[[(1s) -1-[[4- (2-fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl] oxiranecarboxylic acid (0.1 g, 29.6%; compound 12) ) Got. ¹ H NMR (CDCl ₃ ) δ: 2.38 to 2.43 (m, 1H, piperazine ri
ng), 2.83 to 2.93 (m, 2H, piperazine ring), 2.95 to 3.
08 (m, 1H, piperazinering), 3.06 (d, 2H, J = 7.6 Hz,
ph-CH ₂ -C-), 3.20-3.28 (m, 1H, piperazine ring),
3.49-3.66 (m, 1H, piperazine ring), 3.55 (d, 1H, J
= 1.7 Hz, epoxy ring), 3.58 (d, 1H, J = 1.3 Hz, epoxy
ring), 3.67-3.80 (m, 2H, piperazine ring), 4.0-
6.0 (brd, 1H, -COOH), 5.23 (q, 1H, J = 7.9 Hz, -N-CH-
CO), 6.77 to 6.87 (m, 1H, aromatic), 6.93 to 7.09 (m, 3
H, aromatic), 7.20-7.36 (m, 5H, aromatic), 8.23
(d, 1H, J = 8.6 Hz, -NH-).

Example 26 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (4
(2s, 3s) -3-[[-fluorophenyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester in the same manner as in Example 24. [[(1
s) -1-[[4- (4-Fluorophenyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (0.13 g, 69.5
%; Compound 13) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.96 (d, 3H, J = 6.6 H
_{z, -C-CH 3),} 0.99 (d, 3H, J = 6.59 Hz, -C-CH 3), 1.42
(ddd, 1H, J = 14.1,10.5, 3.6 Hz, -C-CH ₂ -C), 1.6 to 1.8
2 (m, 1H, -C- CH-C 2), 1.69 (ddd, 1H, J = 14.5, 10.7,
4.23 Hz, -C-CH ₂ -C), 3.08 to 3.26 (m, 4H, piperazine
ring), 3.55 (d, 1H, J = 1.7 Hz, epoxy ring), 3.60-3.
92 (m, 4H, piperazine ring), 3.62 (d, 1H, J = 1.8 H
z, epoxy ring), 5.08 (ddd, 1H, J = 10.6, 8.6, 3.6 Hz,
-N-CH-CO), 5.2 to 6.4 (brd, 1H, -COOH), 6.84 to 7.0
3 (m, 4H, aromatic), 8.18 (d, 1H, J = 8.6 Hz, -NH-).

Example 27 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[(4-phenyl-1-piperazinyl) carbonyl] -3-methyl]
Using butyl [amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was performed.
(2s, 3s) -3-[[[[(1s) -1-[(4-
Phenyl-1-piperazinyl) carbonyl] -3-methyl] butyl] amino] carbonyl] oxirane carboxylic acid (1.06 g, 29.1%; compound 14) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.96 (d, 3H, J = 6.6 H
z, -C-CH ₃ ), 0.99 (d, 3H, J = 6.26 Hz, -C-CH ₃ ), 1.37
〜1.47 (m, 1H, -C-CH-C ₂ ), 1.64 〜1.80 (m, 2H, -C-CH
₂ -C-), 3.17-3.36 (m, 4H, piperazine ring), 3.55
(d, 1H, J = 1.7 Hz, epoxy ring), 3.62 (d, 1H, J = 1.3 H
z, epoxy ring), 3.70-3.90 (m, 4H, piperazinerin
g), 5.10 (m, 1H, -N-CH-CO), 6.5 to 7.5 (brd, 1H, -CO
OH), 6.87 ~ 6.96 (m, 3H, aromatic), 7.27 ~ 7.33 (m, 2
H, aromatic), 8.20 (d, 1H, J = 8.6 Hz, -NH-).

Example 28 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[(4-dimethylsulfamoyl-1-piperazinyl) carbonyl]
Using (-3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was performed to obtain (2s, 3s) -3-[[[[(1s)-
1-[(4-Dimethylsulfamoyl-1-piperazinyl) carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (1.28 g, 39.0%; compound 15) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.94 (d, 3H, J = 6.3 H
z, -C-CH ₃ ), 0.96 (d, 3H, J = 5.6 Hz, -C-CH ₃ ), 1.36 ~
1.44 (m, 1H, -C-CH-C ₂ ), 1.61 to 1.68 (m, 2H, -C-CH _2-
C-), 2.85 (s, 6H, -N-CH ₃ ), 3.17 to 3.35 (m, 4H, pipe
razine ring), 3.48 to 3.60 (m, 2H, piperazine ring),
3.58 (d, 1H, J = 1.7 Hz, epoxy ring), 3.62 (d, 1H, J
= 1.7 Hz, epoxy ring), 3.70-3.80 (m, 1H, piperazin
e ring), 3.83〜3.95 (m, 1H, piperazine ring), 4.95
5.05.05 (m, 1H, -N-CH-CO), 7.7 8.18.1 (brd, 1H, -COO
H), 7.94 (d, 1H, J = 8.6 Hz, -NH-).

Example 29 (2s, 3s) -1-[[[[(1s) [[4- (4-
Fluorophenyl) -1-piperazinyl] carbonyl]
-2-phenyl] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, (2s, 3
s) -3-[[[[[1s] -1-[[4- (4-methylphenylsulfonyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester And the same operation as in Example 24 was carried out to obtain (2s, 3s) -3-[[[[(1
s) -1-[[4- (4-Methylphenylsulfonyl)
-1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (2.8
g, 70.0%; compound 16) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.90 (d, 3H, J = 6.6 H
z, -C-CH ₃ , 0.93 (d, 3H, J = 6.6 Hz, -C-CH ₃ ), 1.23-1.
33 (m, 1H, -C- CH-C 2), 1.53 ~1.67 (m, 2H, -C-CH 2 -C
-), 2.45 (s, 3H, -ph-CH ₃ ), 2.73 to 2.91 (m, 2H, pipe
razine ring), 3.28 ~ 3.59 (m, 4H, piperazine ring),
3.45 (d, 1H, J = 1.7 Hz, epoxyring), 3.48 (d, 1H, J =
1.7 Hz, epoxy ring), 3.70 to 3.83 (m, 1H, piperazine
ring), 3.98〜4.08 (m, 1H, piperazine ring), 4.85
~ 4.97 (m, 1H, -N-CH-CO), 7.35 (d, 2H, J = 7.9 Hz, a
romatic), 7.63 (d, 2H, J = 8.3 Hz, aromatic), 7.97
(d, 1H, J = 8.6 Hz, -NH-).

Example 30 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
Using (-3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was performed to obtain (2s, 3s) -3-[[[[(1s)-
1-[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (0.41 g, 67.2%; compound 17) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.97 (d, 3H, J = 6.9 H
z, -C-CH ₃ ), 0.98 (d, 3H, J = 7.3 Hz, -C-CH ₃ ), 1.38 〜
1.47 (m, 1H, -C- CH-C 2), 1.65 ~1.77 (m, 2H, -C-CH 2 -
C-), 2.98-3.19 (m, 4H, piperazine ring), 3.55 (d,
1H, J = 1.7 Hz, epoxy ring), 3.61 to 3.77 (m, 2H, pip
erazine ring), 3.64 (d, 1H, J = 1.7 Hz, epoxy ring),
3.77 to 3.89 (m, 1H, piperazine ring), 3.89 to 4.15
(m, 1H, piperazine ring), 5.04-5.18 (m, 1H, -N-CH
-CO), 7.00 to 7.06 (m, 2H, aromatic), 7.21 to 7.28 (m,
1H, aromatic), 7.37 to 7.41 (m, 1H, aromatic), 8.25
(d, 1H, J = 8.9 Hz, -NH-).

Example 31 (2s, 3s) -3-[[[[(1s) -1-[[4-
Instead of (4-fluorophenyl) -1-perazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (3
-Chlorophenyl) -1-piperazinyl] carbonyl]
Using (-3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was performed to obtain (2s, 3s) -3-[[[[(1s)-
1-[[4- (3-Chlorophenyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (0.39 g, 67.2%; compound 18) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.96 (d, 3H, J = 6.6 H
_{z, -C-CH 3),} 0.99 (d, 3H, J = 6.6 Hz, -C-CH 3), 1.36~
1.47 (m, 1H, -C- CH-C 2), 1.64 ~1.80 (m, 2H, -C-CH 2 -
C-), 3.18 to 3.36 (m, 4H, piperazine ring), 3.53 (d,
1H, J = 1.7Hz, epoxy ring), 3.60-3.92 (m, 4H, pip
erazine ring), 3.62 (d, 1H, J = 1.7 Hz, epoxy ring),
5.04-5.12 (m, 1H, -N-CH-CO), 5.5-6.5 (brd, 1H, -C
OOH), 6.77 to 6.82 (m, 1H, aromatic), 6.88 to 6.90 (m,
2H, aromatic), 7.17 to 7.23 (m, 1H, aromatic), 8.21
(d, 1H, J = 8.6 Hz, -NH-).

Example 32 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (4
-Chlorophenyl) -1-piperazinyl] carbonyl]
Using (-3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was performed to obtain (2s, 3s) -3-[[[[(1s)-
1-[[4- (4-Chlorophenyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (0.33 g, 63.4%; compound 19) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.96 (d, 3H, J = 6.6 H
z, -C-CH ₃ ), 0.99 (d, 3H, J = 6.59 Hz, -C-CH ₃ ), 1.36
〜1.47 (m, 1H, -C-CH-C ₂ ), 1.64 〜1.80 (m, 2H, -C-CH
₂ -C-), 3.10 〜 3.31 (m, 4H, piperazine ring), 3.54
(d, 1H, J = 1.7 Hz, epoxy ring), 3.58-3.93 (m, 4H, p
iperazine ring), 3.62 (d, 1H, J = 1.7 Hz, epoxy rin
g), 5.04 to 5.12 (m, 1H, -N-CH-CO), 4.8 to 6.5 (brd,
1H, -COOH), 6.82 to 6.88 (m, 2H, aromatic), 7.21 to 7.
26 (m, 1H, aromatic), 8.18 (d, 1H, J = 8.9 Hz, -NH-).

Example 33 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (4
(2s, 3s) -3-[[-methoxyphenyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester in the same manner as in Example 24. [[(1
s) -1-[[4- (4-Methoxyphenyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (0.49 g, 56.7
%; Compound 20) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.95 (d, 3H, J = 6.3 H
_{z, -C-CH 3),} 0.99 (d, 3H, J = 6.6 Hz, -C-CH 3), 1.38~
1.46 (m, 1H, -C-CH-C ₂ ), 1.63 to 1.80 (m, 2H, -C-CH _2-
C-), 3.05-3.19 (m, 4H, piperazine ring), 3.55 (d,
1H, J = 1.7 Hz, epoxy ring), 3.60-3.90 (m, 4H, pipe
razine ring), 3.62 (d, 1H, J = 1.7 Hz, epoxy ring),
5.06 to 5.13 (m, 1H, -N-CH-CO), 4.8 to 5.8 (brd, 1H,
-COOH), 6.84 to 7.00, (m, 4H, aromatic), 8.14 (d, 1H,
J = 8.6 Hz, -NH-).

Example 34 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
Using (-2-methyl] propyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was carried out to obtain (2s, 3s) -3-[[[[(1s)
-1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] -2-methyl] propyl] amino]
Carbonyl] oxiranecarboxylic acid (1.09 g, 89.5%; compound 21) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 0.98 (d, 3H, J = 6.6 H
z, -C-CH ₃ ), 1.01 (d, 3H, J = 6.6 Hz, -C-CH ₃ , 2.10 (m,
1H, -CH-C ₂ ), 2.98-3.14 (m, 4H, piperazine ring),
3.64 (d, J = 1.6 Hz, 1H, epoxy ring), 3.66 (d, 1H, J =
1.6 Hz, epoxy ring), 3.68 ~ 4.01 (m, 4H, piperazine
ring), 4.98 (dd, 1H, J = 8.9, 5.9 Hz, -N-CH-CO), 7.00
~ 7.06 (m, 2H, aromatic), 7.24 (m, 1H, aromatic),
7.39 (m, 1H, aromatic), 8.29 (d, 1H, J = 8.9, -NH).

Example 35 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
Using s, 3s) -3-[[[[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] methyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was performed. (2s, 3s)
-3-[[[[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] methyl] amino] carbonyl] oxiranecarboxylic acid (1.08 g, 66.5%; compound 2
2) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 3.02 to 3.09 (m, 4H, p
iperazine ring), 3.63 (m, 2H, piperazine ring), 3.6
6 (d, 1H, J = 1.6 Hz, epoxy ring), 3.78 (d, 1H, J = 1.6
Hz, epoxy ring), 3.80 (m, 2H, piperazine ring), 4.
11 (dd, 1H, J = 17.0, 5.4 Hz, -N-CH-CO), 4.33 (dd, 1
H, J = 17.0,5.4 Hz, -N-CH-CO), 6.99 to 7.05 (m, 2H, a
romatic), 7.23 (m, 1H, aromatic), 7.38 (m, 1H, arom
atic), 8.67 (brd, 1H, -NH).

Example 36 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
Same as Example 24 using s, 3s) -3-[[[(1s-1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] ethyl] amino] carbonyl] oxiranecarboxylate ethyl ester. And (2)
s, 3s) -3-[[[(1s-1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] ethyl] amino] carbonyl] oxiranecarboxylic acid (0.87
g, 77.5%; compound 23) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 1.41 (d, 3H, J = 6.8 H
_{z, -C-CH 3),} 2.98~3.13 (m, 4H, piperazine ring), 3.
61 (d, 1H, J = 1.6 Hz, epoxy ring), 3.63 (d, 1H, J = 1.
6 Hz, epoxy ring), 3.67 〜 3.84 (m, 3H, piperazine r
ing), 3.95 (m, 1H, piperazine ring), 5.06 (dq, 1H,
J = 8.4, 6.8 Hz, -N-CH-CO), 7.00 ~ 7.06 (m, 2H, aroma
tic), 7.24 (m, 1H, aromatic), 7.39 (m, 1H, aromati
c), 8.13 (d, 1H, J = 8.4, -NH).

Example 37 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
Using (-2-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was carried out to obtain (2s, 3s) -3-[[[[(1s)-
1-[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] -2-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (1.07 g, 77.4%; compound 24) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 0.92 (t, 3H, J = 7.3 H
z, -C-CH ₃ ), 1.00 (d, 3H, J = 6.8 Hz, -C-CH ₃ ), 1.22
(m, 1H, -CH-C _2- ), 1.54 (m, 1H, -CH-C), 1.84 (m, 1H,
-CH-C), 2.97-3.17 (m, 4H, piperazine ring), 3.60
(d, 1H, J = 1.6 Hz, epoxy ring), 3.64 (d, 1H, J = 1.6 H
z, epoxy ring), 3.65-3.79 (m, 2H, piperazine rin
g), 3.85-4.05 (m, 2H, piperazine ring), 4.98 (dd,
1H, J = 9.2, 6.3 Hz, -N-CH-CO), 7.00-7.06 (m, 2H, a
romatic), 7.24 (m, 1H, aromatic), 7.39 (m, 1H, arom
atic), 8.29 (d, 1H, J = 9.2, -NH).

Example 38 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[2-[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester and the same operation as in Example 24 was performed. And (2s, 3s)
-3-[[[2-[[4- (2-chlorophenyl) -1
-Piperazinyl] carbonyl] ethyl] amino] carbonyl] oxiranecarboxylic acid (0.85 g, 78.9%; compound 2
5) was obtained as colorless crystals.

¹ H NMR (DMSO-d ₆ ) δ: 2.56 (t, 2H, J = 6.9
Hz, -C-CH ₂ -CO), 2.91 to 2.98 (m, 4H, piperazine rin
g), 3.35 (td, 2H, J = 6.9, 5.6 Hz, N-CH ₂ -C), 3.49 (d,
1H, J = 2.0 Hz, epoxy ring), 3.59 (d, 1H, J = 1.6 Hz,
epoxy ring), 3.56 to 3.64 (m, 2H, piperazine ring),
3.80 (m, 2H, piperazine ring), 7.07 (td, 1H, J = 7.
9, 1.7 Hz, aromatic), 7.15 (dd, 1H, J = 7.9, 1.7 Hz,
aromatic), 7.43 (dd, 1H, J = 7.9, 1.7 Hz, aromatic),
7.31 (m, 1H, aromatic), 8.40 (t, 1H, J = 5.6 Hz, -N
H), 13.50 (brd, 1H, -COOH).

Example 39 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[N-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] methyl]-
The same operation as in Example 24 was carried out using ethyl N-methyl] amino] carbonyl] oxiranecarboxylate to give (2s, 3s) -3-[[[[N-[[4- (2-
Chlorophenyl) -1-piperazinyl] carbonyl] methyl] -N-methyl] amino] carbonyl] oxiranecarboxylic acid (1.08 g, 74.8%; compound 26) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 3.01 to 3.10 (m, 6H, p
iperazine ring), 3.27 (s, 3H, -NCH ₃ ), 3.63-3.71
(m, 2H, -N-CH ₂ -CO), 3.75 (d, 1H, J = 1.9 Hz, epoxy ri
ng), 3.78-3.90 (m, 2H, piperazine ring), 4.02 (d,
1H, J = 1.9 Hz, epoxy ring), 6.98-7.05 (m, 2H, aro
matic), 7.24 (m, 1H, aromatic), 7.36 (m, 1H, aromat
I c).

Example 40 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[(2s) -2-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] -1
-Pyrrolidinyl] carbonyl] oxiranecarboxylic acid ethyl ester and the same operation as in Example 24,
(2s, 3s) -3-[[(2s) -2-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
-1-Pyrrolidinyl] carbonyl] oxiranecarboxylic acid (0.94 g, 7.07%; compound 27) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 1.94-2.11 (m, 2H, py
rrolidine ring), 2.17 to 2.30 (m, 2H, pyrrolidine rin
g), 3.06-3.20 (m, 4H, piperazine ring), 3.63-3.76
(m, 2H, piperazine ring), 3.81-3.85 (m, 5H), 4.0
0 (dt, 1H, J = 13.7, 4.4 Hz, pyrrolidine ring), 4.96
(dd, 1H, J = 7.8, 4.3 Hz, pyrrolidine ring), 6.97〜
7.04 (m, 2H, aromatic), 7.24 (m, 1H, aromatic), 7.3
7 (m, 1H, aromatic).

Example 41 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
Using (-2-acetylaminomethylthio] ethyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was carried out to obtain (2s, 3s) -3.
-[[[[(1s) -1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] -2-acetylaminomethylthio] ethyl] amino] carbonyl] oxiranecarboxylic acid (0.88 g, 77.9%; Compound 28) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 2.05 (s, 3H, -COC
H ₃ ), 2.85 (dd, 1H, J = 13.9, 8.3 Hz, -C-CH-S), 2.96
~ 3.16 (m, 5H), 3.69 (d, 1H, J = 1.6 Hz, epoxy ring),
3.78 (d, 1H, J = 1.6 Hz, epoxy ring), 3.71 to 3.89 (m,
4H, piperazine ring), 4.39 (d, 2H, J = 8.3 Hz, -S-CH
₂ -N), 5.21 (m, 1H, -N-CH-CO), 6.97 to 7.02 (m, 2H,
aromatic), 7.22 (m, 1H, aromatic), 7.36 (m, 1H, aro
matic), 7.80 (d, 1H, J = 8.3, -NH), 9.00 (brd, 1H, -N
H).

Example 42 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
Example 24 using -3-methylthio] propyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester
The same operation as described above is performed, and (2s, 3s) -3-[[[[(1
s) -1-[[4- (2-Chlorophenyl) -1-piperazinyl] carbonyl] -3-methylthio] propyl]
Amino] carbonyl] oxiranecarboxylic acid (1.17 g,
80.7%; compound 29) was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 1.98 (dd, 2H, J = 6.9,
6.6 Hz, -CH-CC), 2.12 (s, 3H, -SCH ₃ ), 2.57 (dt, 2
H, J = 6.9, 2.3 Hz, -C-CH ₂ -CS), 3.05 to 3.19 (m, 4H,
piperazine ring), 3.63 (d, 1H, J = 1.9 Hz, epoxy rin
g), 3.65 (d, 1H, J = 1.9 Hz, epoxy ring), 3.71-3.94
(m, 4H, piperazine ring), 5.26 (m, 1H, -N-CH-CO),
7.00 ~ 7.06 (m, 2H, aromatic), 7.24 (m, 1H, aromati
c), 7.38 (m, 1H, aromatic), 8.18 (d, 1H, J = 8.6, -N
H).

Example 43 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[[(1s) -1-[[4- (2
-Chlorophenyl) -1-piperazinyl] carbonyl]
-3-carbamoyl] propyl] amino] carbonyl]
Example 2 using ethyl oxiranecarboxylate
Perform the same operation as in (4), and (2s, 3s) -3-
[[[[[(1s) -1-[[4- (2-chlorophenyl) -1-piperazinyl] carbonyl] -3-carbamoyl] propyl] amino] carbonyl] oxiranecarboxylic acid (0.14 g, 74.5%; compound 30) Was obtained as colorless crystals.

¹ H NMR (CDCl ₃ ) δ: 1.85 (brd, 2H, -N
H ₂ ), 2.14 (m, 1H, -C-CH-C-CO-), 2.36 to 2.53 (m, 3H,
-CH-CH ₂ -C-CO-), 2.89 ~ 3.06 (m, 4H, piperazine rin
g), 3.57 to 3.79 (m, 6H, piperazine and epoxy ring),
5.02 (m, 1H, -N-CH-CO), 6.96 to 7.02 (m, 2H, aromat
ic), 7.21 (m, 1H, aromatic), 7.37 (m, 1H, aromati
c), 7.88 (brd, 1H, -NH).

Example 44 (2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Fluorophenyl) -1-piperazinyl] carbonyl] -2-phenyl] ethyl] amino] carbonyl]
Instead of oxiranecarboxylic acid ethyl ester, (2
s, 3s) -3-[[[(1s) -1-[[4- (4-
Fluorophenyl) -1-piperazinyl] carbonyl]
Using (1-cyclopentyl] amino] carbonyl] oxiranecarboxylic acid ethyl ester, the same operation as in Example 24 was carried out to obtain (2s, 3s) -3-[[[(1s) -1
-[[4- (4-Fluorophenyl) -1-piperazinyl] carbonyl] -1-cyclopentyl] amino] carbonyl] oxiranecarboxylic acid (0.52 g, 55.6%; compound 31) was obtained as colorless crystals.

¹ H NMR (DMSO-d ₆ ) δ: 1.61 (m, 4H, cyclo
pentyl), 1.87 (m, 2H, cyclopentyl), 2.22 (m, 2H, cy
clopentyl), 2.97 (m, 4H, piperazine), 3.45 (d, 1H,
J = 1.6 Hz, epoxy ring), 3.58 (d, 1H, J = 2.1 Hz, epoxy
ring), 3.60 (m, 4H, piperazine ring), 6.95-7.20
(m, 4H, aromatic), 8.89 (s, 1H, -NH), 13.4 (brd, 1
H, -COOH).

Example 45 (2s, 3s) -3- obtained in Example 29
[[[[[(1s) -1-[[4- (4-methylphenylsulfonyl) -1-piperazinyl] carbonyl] -3-
Methyl] butyl] amino] carbonyl] oxiranecarboxylic acid (0.935 g, 2 mmol) in dichloromethane (15 ml)
Add o-benzylhydroxylamine (0.638 g, 4.0
mmol) and N-methylmorpholine (0.405 g, 4.0 mmol)
And dicyclohexylcarbodiimide (0.619
g, 3.0 mmol) in dichloromethane (5 ml) was added dropwise under ice cooling. After stirring at room temperature for 24 hours, the precipitate was removed by filtration, washed with dichloromethane (20 ml), combined with the filtrate, and washed with water. The organic layer was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and the residue was subjected to silica gel chromatography. Elution with ethyl acetate: hexane (2: 1) gave (2s, 3s) -3-[[[[(1s) -1-
[[4- (4-Methylphenylsulfonyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarboxylic acid benzyloxyamide (0.86 g, 75.1%; compound 32) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.84 (d, 3H, J = 6.2 H
z, -C-CH ₃ ), 0.91 (d, 3H, J = 6.5 Hz, -C-CH ₃ ), 1.23 ~
1.31 (m, 1H, -C-CH ₂ -C), 1.36 to 1.58 (m, 2H, -C-CH-C,
-C-CH-C ₂ ), 2.43 (s, 3H, -ph-CH ₃ ), 2.72 to 2.86 (m, 2
H, piperazine ring), 3.14-3.27 (m, 2H, piperazin
ering), 3.31 to 3.51 (m, 2H, piperazine ring), 3.40
(d, 1H, J = 1.4 Hz, epoxy ring), 3.43 (d, 1H, J = 1.7
Hz, epoxy ring), 3.63 〜 3.74 (m, 1H, piperazine rin
g), 3.84 to 3.98 (m, 1H, piperazine ring), 4.80 to 4.90
(m, 1H, -N-CH -CO), 4.87 (s, 3H, -O-CH 2 -ph), 7.30~
7.40 (m, 8H, aromatic, -NH-), 7.56-7.66 (m, 2H, a
romatic), 9.05 (s, 1H, -NH-).

Example 46 (2s, 3s) -3- obtained in Example 45
[[[[[(1s) -1-[[4- (4-methylphenylsulfonyl) -1-piperazinyl] carbonyl] -3-
A catalytic amount of palladium carbon was added to a solution of methyl] butyl] amino] carbonyl] oxiranecarboxylate benzyloxyamide (0.57 g, 1 mmol) in methanol (25 ml) to perform catalytic hydrogen reduction. After completion of the reaction, palladium carbon was removed by filtration, the filtrate was concentrated, and the residue was subjected to silica gel chromatography. Elution with ethyl acetate,
(2s, 3s) -3-[[[[(1s) -1-[[4-
(4-Methylphenylsulfonyl) -1-piperazinyl] carbonyl] -3-methyl] butyl] amino] carbonyl] oxiranecarbohydroxamic acid (0.18 g, 3
7.3%; compound 33) was obtained.

¹ H NMR (CDCl ₃ ) δ: 0.84 (d, 3H, J = 5.9 H
_{z, -C-CH 3),} 0.90 (d, 3H, J = 5.9 Hz, -C-CH 3), 1.24~1.
33 (m, 1H, -C-CH ₂ -C), 1.50 to 1.64 (m, 2H, -C-CH-C,-
C-CH-C ₂ ), 2.42 (s, 3H, -ph-CH ₃ ), 2.90 to 3.20 (m, 4
H, piperazine ring), 3.44-3.80 (m, 3H, piperazine
ring), 3.51 (s, 1H, epoxy ring), 3.68 (s, 1H, epoxy
ring), 4.56〜4.66 (m, 1H, piperazine ring), 4.76〜
4.90 (m, 1H, -N-CH-CO), 7.33 (d, 2H, J = 7.8 Hz, arom
atic), 7.62 (dd, 2H, J = 7.8, 1.7 Hz, aromatic), 7.84
〜7.94 (brd, 1H, -NH-) 9.80 〜10.40 (brd, 1H, -O
H).

Example 47 (Tablet) Compound 23 88 mg Starch 17 mg Magnesium stearate 3 mg The above ingredients are molded into tablets by a conventional method using one tablet as a material. Dragees may be attached as necessary.

Example 48 (Capsule) Compound 18 50 mg Lactose 100 mg Starch 30 mg Magnesium stearate 10 mg The above components are mixed, and the mixture is filled in a gelatin capsule as a material for one capsule.

Example 49 (Injection) Compound 21 2.5 mg Sodium chloride 900 mg 1N Sodium hydroxide Appropriate amount Distilled water for injection Total amount 100 ml The above components are mixed by a conventional method to prepare an injection.

Example 50 (Eye drops) Compound 18 50 mg Boric acid 700 mg Borax proper amount Sodium chloride 500 mg Sodium edetate 0.05 mg Benzalkonium chloride 0.005 mg Sterilized purified water Total amount 100 ml or more are mixed by a conventional method. To be used as eye drops.

Example 51 N- (2-Naphthalenesulfonyl) -L-valyl-L-
Leucinal valine (11.5 g) in 1 M sodium hydroxide aqueous solution 1
In 100 ml of purified water, and 200 ml of purified water and 100 ml of tetrahydrofuran were added thereto.
100 ml of a 1 M aqueous sodium hydroxide solution and 100 ml of a solution of 2-naphthalenesulfonyl chloride (18.5 g) in tetrahydrofuran were simultaneously added dropwise. This solution was stirred at room temperature for 24 hours to react. After the reaction is completed, the reaction solution is adjusted to pH
Adjusted to 2-3 and extracted with ethyl acetate. The extract was washed with diluted hydrochloric acid and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution to obtain 12.8 g of N- (2-naphthalenesulfonyl) -L-valine as white crystals.

N- (2-naphthalenesulfonyl) -L-
Valine (12.0 g) and N-hydroxysuccinimide (5.4 g) are dissolved in 200 ml of tetrahydrofuran, and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (9 .0
200 ml of a dichloromethane solution of g) were slowly added. The solution was stirred at room temperature for about 4 hours to react. After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a mixed solution of hexane and ethyl acetate, and 14.1 g of N- (2-naphthalenesulfonyl) -L-valine N-hydroxysuccinimide ester was obtained.
Was obtained as white crystals.

N- (2-naphthalenesulfonyl) -L-
Valine N-hydroxysuccinimide ester (1.8
g) and leucinol (0.63 g) in dichloromethane 1
Then, triethylamine (0.68 g) was added while stirring at room temperature. The solution was stirred for 2 hours to react. After the completion of the reaction, the resultant was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. The dichloromethane was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution, and N- (2-naphthalenesulfonyl) -L-valyl-L-leucinol 1.3 was obtained.
g were obtained as white crystals.

N- (2-naphthalenesulfonyl) -L-
Valyl-L-leucinol (1.3 g) was dissolved in 20 ml of dimethylsulfoxide and 10 ml of dichloromethane, and triethylamine (1.9 g) was added. While stirring this solution at room temperature, 20 ml of a dimethyl sulfoxide solution of a sulfur trioxide pyridine complex (2.0 g) was added, and the mixture was further stirred for 2 hours. After completion of the reaction, ethyl acetate was added, and diluted hydrochloric acid,
After washing with saturated aqueous sodium hydrogen carbonate solution and saturated saline,
It was dehydrated with anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution.
0.98 g of (2-naphthalenesulfonyl) -L-valyl-L-leucinal (compound 34) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.42 (d, 3
H, J = 6.3Hz), 0.55 (d, 3H, J = 6.3Hz), 0.84 (d, 3H, J =
6.6Hz), 0.88 (d, 3H, J = 6.6Hz), 0.93-1.12 (m, 2H),
1.14-1.28 (m, 1H), 1.82-2.00 (m, 1H), 3.63-3.72 (m,
2H), 7.62-8.40 (m, 9H), 9.02 (s, 1H). Anal. (C ₂₁ H ₂₈ N ₂ O ₄ S) C, H, N.

Example 51-2 Using 4-fluorobenzenesulfonyl chloride in place of 2-naphthalenesulfonyl chloride of Example 51,
By performing the same operation as in Example 51, N- (4-fluorophenylsulfonyl) -L-valyl-L-leucinal (compound 35) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.74 (d, 3
H, J = 5.9Hz), 0.80 (d, 6H, J = 6.4Hz), 0.85 (d, 3H, J =
6.8Hz), 1.14-1.46 (m, 3H), 1.81-1.93 (m, 1H), 3.56-
3.62 (dd, 1H, J = 6.6, 9.5Hz), 3.80-3.88 (m, 1H), 7.3
3-7.42 (m, 2H), 7.79-7.86 (m, 2H), 7.96 (d, 1H, J =
9.8Hz), 8.27 (d, 1H, J = 7.3Hz), 9.14 (s, 1H). Anal. (C ₁₇ H ₂₅ FN ₂ O ₄ S) C, H, N.

Example 51-3 N- (4-chlorophenylsulfonyl) -L-valyl was prepared in the same manner as in Example 51 except that 4-chlorobenzenesulfonyl chloride was used in place of 2-naphthalenesulfonyl chloride in Example 51. -L-Leucinal (compound 36) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.74 (d, 3
H, J = 5.9Hz), 0.82 (d, 6H, J = 6.8Hz), 0.88 (d, 3H, J =
6.3Hz), 1.15-1.46 (m, 1H), 3.61 (dd, 1H, J = 6.8, 9.3
Hz), 3.82-3.90 (m, 1H), 7.56-7.63 (m, 2H), 7.44-7.7
9 (m, 2H), 8.03 (d, 1H, J = 9.3Hz), 8.26 (d, 1H, J =
7.3Hz), 9.15 (s, 1H). Anal. (C ₁₇ H ₂₅ ClN ₂ O ₄ S) C, H, N.

Example 51-4 Example 5 was repeated using p-toluenesulfonyl chloride in place of 2-naphthalenesulfonyl chloride in Example 51.
1-N- (4-methylphenylsulfonyl) -L-valyl-L-leucinal (Compound 37)
Was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.72-0.90
(m, 12H), 1.18-1.45 (m, 3H), 1.79-1.91 (m, 1H), 2.3
6 (s, 3H), 3.57 (t, 1H, J = 7.7Hz), 3.77-3.84 (m, 1
H), 7.32 (d, 2H), 7.62-7.70 (m, 2H), 7.76 (d, 1H, J
= 8.3Hz), 8.26 (d, 1H, J = 6.8Hz), 9.07 (s, 1H). Anal. (C 18 H 28 N 2 O 4 S) C, H, N.

Example 51-5 N- (2-Naphthalenesulfonyl) -L-tert-leucyl-L-leucinal was prepared in the same manner as in Example 51 except that tert-leucine was used in place of valine in Example 51. (Compound 38) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.35 (d, 3
H, J = 6.4Hz), 0.46 (d, 3H, J = 6.4Hz), 0.78-0.95 (m, 2
H), 0.95 (s, 9H), 1.08-1.20 (m, 1H), 3.45-3.55 (m,
1H), 3.67 (d, 1H, J = 10.3Hz), 7.62-7.72 (m, 2H), 7.8
2-7.86 (m, 1H), 7.97-8.10 (m, 4H), 8.17 (d, 1H, J =
6.4Hz), 8.29 (m, 1H), 8.91 (s, 1H). Anal. (C ₂₂ H ₃₀ N ₂ O ₄ S) C, H, N.

Example 51-6 Instead of 2-naphthalenesulfonyl chloride of Example 51, 4-fluorobenzenesulfonyl chloride was used.
N- (4-Fluorophenylsulfonyl)
-D-Valyl-L-leucinal (compound 39) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.78 (d, 3
H, J = 6.3Hz), 0.82 (d, 3H, J = 6.9Hz), 0.83 (d, 6H, J =
6.3Hz), 1.24-1.50 (m, 3H), 1.80-1.92 (m, 1H), 3.62
(s br, 1H), 3.84-3.92 (m, 1H), 7.32-7.41 (m, 2H),
7.79 (m, 3H), 8.33 (d, 1H, J = 6.9Hz), 8.96 (s, 1H). Anal. (C ₂₂ H ₃₀ N ₂ O ₄ S) C, H, N.

Example 51-7 Instead of 2-naphthalenesulfonyl chloride of Example 51, 4-fluorobenzenesulfonyl chloride was used.
N- (4-fluorophenylsulfonyl) -L-norleucyl-L-leucinal (compound 4) was prepared in the same manner as in Example 51 except that norleucine was used instead of valine.
0) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.74-0.90
(m, 9H), 1.07-1.59 (m, 9H), 3.76 (t, 1H, J = 5.4Hz),
3.84-3.91 (m, 1H), 7.34-7.45 (m, 2H), 7.79-8.07 (m,
3H), 8.29 (d, 1H, J = 7.3Hz), 9.18 (s, 1H). Anal. (C ₂₂ H ₃₀ N ₂ O ₄ S) C, H, N.

Example 51-8 In place of 2-naphthalenesulfonyl chloride in Example 51, 4-fluorobenzenesulfonyl chloride was used.
N- (4-fluorophenylsulfonyl) was obtained in the same manner as in Example 51, using norvaline instead of valine.
-L-Norvalyl-L-leucinal (Compound 41) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.69-0.85
(m, 9H), 1.14-1.66 (m, 7H), 3.78 (t, 1H, J = 6.3Hz),
3.84-3.92 (m, 1H), 7.34-7.42 (m, 2H), 7.79-8.02
(m, 3H), 8.28 (d, 1H, J = 7.3Hz), 9.18 (s, 1H) .; Anal. (C ₂₂ H ₃₀ N ₂ O ₄ S) C, H, N.

Example 52 N- (2-Naphthalenesulfonyl) -L-tert-b
Isyl-L-phenylalaninal tert-leucine (13.1 g) was dissolved in 100 ml of a 1 M aqueous sodium hydroxide solution, and purified water 200 m
and 100 ml of tetrahydrofuran were added, and 100 ml of a 1M aqueous sodium hydroxide solution and 100 ml of a tetrahydrofuran solution of 2-naphthalenesulfonyl chloride (20.4 g) were simultaneously added dropwise with stirring under ice cooling. This solution was stirred at room temperature for 24 hours to react. After completion of the reaction, the reaction solution was adjusted to pH 2-3 and extracted with ethyl acetate. The extract was washed with diluted hydrochloric acid and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution to give N- (2-naphthalenesulfonyl) -L-tert-leucine.
5 g were obtained as white crystals.

N- (2-naphthalenesulfonyl) -L-
Tert-leucine (16.0 g) and N-hydroxysuccinimide (6.9 g) were added to tetrahydrofuran 200
of 1-ethyl-3- while stirring under ice-cooling.
200 ml of a dichloromethane solution of (3-dimethylaminopropyl) carbodiimide hydrochloride (11.5 g) was slowly added. The solution was stirred at room temperature for about 12 hours to react. After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution to obtain 18.3 g of N- (2-naphthalenesulfonyl) -L-tert-leucine N-hydroxysuccinimide ester as white crystals. .

N- (2-naphthalenesulfonyl) -L-
tert-leucine N-hydroxysuccinimide ester (1.8 g) and phenylalaninol (1.0 g)
Was added to 50 ml of dichloromethane, and triethylamine (0.86 g) was added with stirring at room temperature. The solution was stirred for 2 hours to react. After the completion of the reaction, the resultant was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Dichloromethane was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution.
1.6 g of-(2-naphthalenesulfonyl) -L-tert-leucyl-L-phenylalaninol was obtained as white crystals.

N- (2-naphthalenesulfonyl) -L-
tert-leucyl-L-phenylalaninol (1.
6 g) was dissolved in dimethyl sulfoxide (20 ml) and dichloromethane (10 ml), triethylamine (2.1 g) was added, and the mixture was stirred at room temperature with stirring to obtain a sulfur trioxide pyridine complex (2.
15 g of a dimethyl sulfoxide solution of 2 g) were added.
The solution was stirred for 2 hours to react. After completion of the reaction, ethyl acetate was added, and the mixture was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution, and N- (2-naphthalenesulfonyl)-
1.1 g of L-tert-leucyl-L-phenylalaninal (compound 42) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.86 (s, 9
H), 2.26-2.40 (m, 1H), 2.63-2.77 (m, 1H), 3.56 (dd,
1H, J = 6.8, 13.2Hz), 3.63-3.68 (m, 1H), 6.87-6.90
(m, 1H), 6.99-7.03 (m, 1H), 7.11-7.22 (m, 3H), 7.60
-7.72 (m, 2H), 7.80-7.87 (m, 1H), 7.92-8.19 (m, 4
H), 8.35 (d, 1H, J = 6.8Hz), 8.40-8.43 (m, 1H), 8.63
(s, 1H). Anal. (C ₂₅ H ₂₈ N ₂ O ₄ S) C, H, N.

Example 52-2 Using 4-fluorobenzenesulfonyl chloride instead of 2-naphthalenesulfonyl chloride of Example 52,
Example 5 using valine instead of tert-leucine
By performing the same operation as in 2, N- (4-fluorophenylsulfonyl) -L-valyl-L-phenylalaninal (compound 43) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.76 (d, 3
H, J = 6.4Hz), 0.77 (d, 3H, J = 6.4Hz), 1.69-1.86 (m, 1
H), 2.67 (dd, 1H, J = 8.8,14.2Hz), 3.02 (dd, 1H, J =
5.1, 14.2Hz), 3.56 (dd, 1H, J = 6.4, 9.3Hz), 3.99-4.0
7 (m, 1H), 7.12-7.29 (m, 7H), 7.72-7.84 (m, 2H),
7.92 (d, 1H, J = 9.3Hz), 8.44 (d, 1H, J = 6.8Hz), 9.07
(s, 1H). Anal. (C ₂₀ H ₂₃ FN ₂ O ₄ S) C, H, N.

Example 52-3 N- (2-naphthalenesulfonyl) -L-valyl-L-phenylalaninal was prepared in the same manner as in Example 52 except that valine was used instead of tert-leucine. (Compound 44) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.63 (d, 3
H, J = 6.6Hz), 0.76 (d, 3H, J = 6.6Hz), 1.68-1.82 (m, 1
H,), 2.40-2.92 (m, 1H), 3.64 (dd, 1H, J = 6.6, 9.2H
z), 3.97-3.87 (m, 1H), 6.95-7.02 (m, 2H), 7.10-7.23
(m, 3H), 7.62-7.82 (m, 3H), 7.94-8.10 (m, 4H), 8.
36-8.43 (m, 2H), 8.86 (s, 1H). Anal. (C ₂₄ H ₂₆ N ₂ O ₄ S) C, H, N.

Example 52-4 Using 4-chlorobenzenesulfonyl chloride instead of 2-naphthalenesulfonyl chloride of Example 52,
Example 52 using valine instead of ert-leucine
By the same operation as described above, N- (4-chlorophenylsulfonyl) -L-valyl-L-phenylalaninal (compound 45) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.77 (d, 3
H, J = 6.8Hz), 0.79 (d, 3H, J = 6.8Hz), 1.70-1.87 (m, 1
H), 2.67 (dd, 1H, J = 8.8,14.2Hz), 3.01 (dd, 1H, J =
5.4, 14.2Hz), 3.60 (dd, 1H, J = 6.4, 9.3Hz), 4.00-4.0
7 (m, 1H), 7.12-7.32 (m, 5H), 7.50-7.60 (m, 2H),
7.68-8.00 (m, 2H), 7.98 (d, 1H, J = 9.3Hz), 8.44 (d,
1H, J = 6.8Hz), 9.09 (s, 1H). Anal. (C ₂₀ H ₂₃ ClN ₂ O ₄ S) C, H, N.

Example 52-5 p-Toluenesulfonyl chloride was used in place of 2-naphthalenesulfonyl chloride of Example 52, and tert
-Using valine instead of leucine and performing the same operation as in Example 52, N- (4-methylphenylsulfonyl)-
L-valyl-L-phenylalaninal (compound 46)
Was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.74 (d, 6
(H, J = 6.4Hz), 1.71-1.81 (m, 1H), 2.33 (s, 3H), 2.65
(dd, 1H, J = 8.8, 14.2), 2.99 (dd, 1H, J = 5.4, 14.2),
3.55 (dd, 1H, J = 6.4, 9.3Hz), 3.97-4.05 (m, 1H), 7.1
1-7.37 (m, 7H), 7.59-7.66 (m, 2H), 7.73 (d, 1H, J =
9.3Hz), 8.41 (d, 1H, J = 6.8Hz), 8.99 (s, 1H). Anal. (C ₂₁ H ₂₆ N ₂ O ₄ S) C, H, N.

Example 52-6 1- (2-Naphthalenesulfonylamino) cyclohexanecarbonyl-L was prepared in the same manner as in Example 52 except that 1-aminocyclohexanecarboxylic acid was used in place of tert-leucine. -Phenylalaninol (compound 47) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 1.12 (sb
r, 6H), 1.65 (s br, 4H), 2.28 (dd, 1H, J = 8.6, 14.2H
z), 3.06 (dd, 1H, J = 5.3,14.2Hz), 4.07-4.14 (m, 1
H), 7.16-7.29 (m, 5H), 7.63-7.72 (m, 2H), 7.86-7.72
(m, 2H), 7.98-8.15 (m, 4H), 8.41 (s, 1H), 9.29 (s,
1H). Anal. (C ₂₆ H ₂₈ N ₂ O ₄ S) C, H, N.

Example 53 N- (4-Chlorophenylsulfonyl) -L-valyl-
L-tryptophanal valine (13.1 g) was added to a 1 M aqueous sodium hydroxide solution 1
And then add 250 ml of purified water and 100 ml of tetrahydrofuran.
100 ml of an aqueous M sodium hydroxide solution and 100 ml of a tetrahydrofuran solution of 4-chlorobenzenesulfonyl chloride (19.0 g) were alternately added in five portions. This solution was stirred at room temperature for 24 hours to react. After completion of the reaction, the reaction solution was adjusted to pH 2-3 and extracted with ethyl acetate. The extract was washed with diluted hydrochloric acid and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a mixed solution of hexane-ethyl acetate to obtain 13.6 g of N- (4-chlorophenylsulfonyl) -L-valine as white crystals.

N- (4-chlorophenylsulfonyl)-
L-valine (13.5 g) and N-hydroxysuccinimide (6.4 g) are dissolved in 200 ml of tetrahydrofuran, and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride is stirred under ice-cooling. (10.
200 ml of a 6 g) dichloromethane solution was slowly added. The solution was stirred at room temperature for about 12 hours to react. After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate.
Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution to obtain 14.3 g of N- (4-chlorophenylsulfonyl) -L-valine N-hydroxysuccinimide ester as white crystals.

N- (4-chlorophenylsulfonyl)-
L-valine N-hydroxysuccinimide ester (1.5 g) and L-tryptophanol (0.88 g)
Was added to 100 ml of dichloromethane, and triethylamine (1.2 g) was added with stirring at room temperature. The solution was stirred for 2 hours to react. After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure,
The residue was washed with a hexane-ethyl acetate mixture, and N- (4-
1.6 g of (chlorophenylsulfonyl) -L-valyl-L-tryptophanol were obtained as white crystals.

N- (4-chlorophenylsulfonyl)-
L-valyl-L-tryptophanol (1.5 g) was added to 20 ml of dimethyl sulfoxide and 15 ml of dichloromethane.
And triethylamine (2.0 g) was added thereto, and while stirring at room temperature, 20 ml of a dimethylsulfoxide solution of a sulfur trioxide pyridine complex (2.1 g) was added, and the mixture was further stirred for 1 hour. After completion of the reaction, ethyl acetate was added, and the mixture was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent is distilled off under reduced pressure,
The residue was purified using a preparative TLC plate (developing solvent: hexane-ethyl acetate, 1: 1), and 0.10 g of N- (4-chlorophenylsulfonyl) -L-valyl-L-tryptophanal (compound 48) was used. Was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.81 (d, 3
H, J = 6.8Hz), 0.82 (d, 3H, J = 6.4Hz), 1.77-1.91 (m, 1
H), 2.82 (dd, 1H, J = 7.8, 15.1Hz), 3.07 (dd, 1H, J =
5.9, 15.1Hz), 3.65 (dd, 1H, J = 6.8, 9.3Hz), 4.06-4.1
4 (m, 1H), 6.96-7.69 (m, 9H), 7.99 (d, 1H, J = 9.8H
z), 8.41 (d, 1H, J = 6.4Hz), 9.21 (s, 1H), 10.92 (s,
1H) .; Anal. (C ₂₂ H ₂₄ ClN ₃ O ₄ S) C, H, N.

Example 53-2 N- (4-fluorophenylsulfonyl) -L was prepared in the same manner as in Example 53 except that 4-chlorobenzenesulfonyl chloride was used instead of 4-chlorobenzenesulfonyl chloride. -Valyl-L-tryptophanal (compound 49) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.80 (d, 3
H, J = 6.8Hz), 0.81 (d, 3H, J = 6.8Hz), 1.76-1.88 (m, 1
H), 2.82 (dd, 1H, J = 8.1,15.1Hz), 3.06 (dd, 1H, J =
6.1, 15.1Hz), 3.63 (dd, 1H, J = 6.8, 9.3Hz), 4.04-4.1
2 (m, 1H), 6.98-7.56 (m, 7H), 7.68-7.76 (m, 2H),
7.93 (d, 1H, J = 9.3Hz), 8.41 (d, 1H, J = 6.4Hz), 9.19
(s, 1H), 10.92 (s, 1H). Anal. (C ₂₂ H ₂₄ FN ₃ O ₄ S) C, H, N.

Example 53-3 The same operation as in Example 53 was carried out except that 2-naphthalenesulfonyl chloride was used instead of 4-chlorobenzenesulfonyl chloride and 1-aminocyclohexanecarboxylic acid was used instead of valine. 1- (2-Naphthalenesulfonylamino) cyclohexanecarbonyl-L
-Tryptophanal (compound 50) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 1.17 (sb
r, 6H), 1.72 (s br, 4H), 2.97-3.16 (m, 2H), 4.10-4.
17 (m, 1H), 6.95-7.22 (m, 3H), 7.33 (d, 1H, J = 8.3H
z), 7.48 (d, 1H, J = 7.6Hz), 7.61-7.72 (m, 2H), 7.83
-8.14 (m, 6H), 8.41 (s, 1H), 10.89 (s, 1H). Anal. (C ₂₈ H ₂₉ N ₃ O ₄ S) C, H, N.

Example 53-4 Example 53 was repeated using 2-naphthalenesulfonyl chloride in place of 4-chlorobenzenesulfonyl chloride and tert-leucine in place of valine.
And N- (2-naphthalenesulfonyl)
-L-tert-Leucyl-L-tryptophanal (Compound 51) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.89 (s, 9
H), 2.43 (dd, 1H, J = 6.8, 15.1Hz), 2.68 (dd, 1H, J =
7.3, 15.1Hz), 3.64-3.75 (m, 2H), 6.93-7.16 (m, 3H),
7.19 (d, 1H, J = 7.8Hz), 7.32 (d, 1H, J = 8.3Hz), 7,58
-7.67 (m, 2H), 7.76-7.80 (m, 2H), 7.88-8.01 (m, 3
H), 8.05-8.09 (m, 1H), 8.37 (d, 1H, J = 6.4Hz), 8.43
(m, 1H), 8.83 (s, 1H), 10.80 (s, 1H) .; Anal. (C ₂₇ H ₂₉ N ₃ O ₄ S) C, H, N.

Example 54 N- (4-Fluorophenylsulfonyl) -L-valyl
-L-cyclohexylalaninal valine (11.5 g) was added to a 1 M aqueous sodium hydroxide solution 1
In 200 ml of purified water, and 200 ml of purified water and 100 ml of tetrahydrofuran were added.
100 ml of M sodium hydroxide aqueous solution and 100 ml of a tetrahydrofuran solution of 4-fluorobenzenesulfonyl chloride (17.5 g) were simultaneously added dropwise. This solution was stirred at room temperature for 24 hours to react. After completion of the reaction, the reaction solution was adjusted to pH 2-3 and extracted with ethyl acetate. The extract was washed with diluted hydrochloric acid and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution to obtain 15.5 g of N- (4-fluorophenylsulfonyl) -L-valine as white crystals.

N- (4-fluorophenylsulfonyl)
-L-Valine (12.0 g) and N-hydroxysuccinimide (7.6 g) are dissolved in 200 ml of tetrahydrofuran, and 1-ethyl-3- (3-
Dimethylaminopropyl) carbodiimide hydrochloride (1
A solution of 2.6 g) in dichloromethane was slowly added. The solution was stirred at room temperature for about 4 hours to react. After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate.
Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution to obtain 14.1 g of N- (4-fluorophenylsulfonyl) -L-valine N-hydroxysuccinimide ester as white crystals.

N- (4-fluorophenylsulfonyl)
-L-valine N-hydroxysuccinimide ester (1.5 g) and (S) -2-amino-3-cyclohexyl-1-propanol hydrochloride (1.5 g) are added to 80 ml of dichloromethane, and the mixture is stirred at room temperature while stirring. Triethylamine (2.0 g) was added. The solution is stirred for 2 hours,
Reacted. After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a mixed solution of hexane-ethyl acetate to obtain 1.4 g of N- (4-fluorophenylsulfonyl) -L-valyl-L-cyclohexylalaninol as white crystals.

N- (4-fluorophenylsulfonyl)
-L-valyl-L-cyclohexylalaninol (1.
3g) was dissolved in 20 ml of dimethyl sulfoxide solution and 10 ml of dichloromethane, and triethylamine (1.9 g) was dissolved.
Was added. While stirring this solution at room temperature, a solution of sulfur trioxide pyridine complex (2.0 g) in dimethyl sulfoxide 1
0 ml was added and stirring was continued for another hour. After completion of the reaction, ethyl acetate was added, and the mixture was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified using a preparative TLC plate (developing solvent: hexane-ethyl acetate, 1: 1) and crystallized from isopropyl ether.
N- (4-Fluorophenylsulfonyl) -L-valyl-L-cyclohexylalaninal (Compound 52)
37 g were obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.74-1.61
(m, 13H), 0.82 (d, 3H, J = 10.9Hz), 0.84 (d, 3H, J = 1
0.9Hz), 1.80-1.93 (m, 1H), 3.53-3.66 (m, 1H), 3.77-
3.85 (m, 1H), 7.32-7.42 (m, 2H), 7.79-7.87 (m, 2H),
7.96 (d, 1H, J = 8.9Hz), 8.29 (d, 1H, J = 6.6Hz), 9.1
0 (s, 1H). Anal. (C ₂₀ H ₂₉ FN ₂ O ₄ S) C, H, N.

Example 54-2 In place of 4-fluorobenzenesulfonyl chloride in Example 54, 2-naphthalenesulfonyl chloride was used.
By the same operation as in Example 54, N- (2-naphthalenesulfonyl) -L-valyl-L-cyclohexylalaninal (compound 53) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.52-0.82
(m, 13H), 0.82 (d, 3H, J = 6.6Hz), 0.84 (d, 3H, J = 5.6
Hz), 1.81-1.99 (m, 1H), 3.63-3.69 (m, 2H), 7.80 (d
d, 1H, J = 1.9, 8.8Hz), 8.00-8.11 (m, 4H), 8.26 (d, 1
H, J = 6.6Hz), 8.39 (m, 1H), 8.96 (s, 1H). Anal. (C ₂₄ H ₃₂ N ₂ O ₄ S) C, H, N.

Example 54-3 N- (4-chlorophenylsulfonyl) -L- was obtained in the same manner as in Example 54, except that 4-chlorophenylsulfonyl chloride was used instead of 4-fluorobenzenesulfonyl chloride. Valyl-L-cyclohexylalaninal (Compound 54) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.74-1.61
(m, 13H), 0.82 (d, 3H, J = 10.2Hz), 0.85 (d, 3H, J = 1
0.5Hz), 1.89-1.93 (m, 1H), 3.58-3.63 (m, 1H), 3.77-
3.85 (m, 1H), 7.58-7.63 (m, 2H), 7.75-7.80 (m, 2H),
8.05 (d, 1H, J = 7.3Hz), 8.40 (d, 1H, J = 6.6Hz), 9.1
1 (s, 1H). Anal. (C ₂₀ H ₂₉ Cl-N ₂ O ₄ S) C, H, N.

Example 55 N- (4-Fluorophenylsulfonyl) -D-valyl
-D-leucineal D-valine (6.6 g) was dissolved in 50 ml of a 1 M aqueous sodium hydroxide solution, and 200 ml of purified water and 100 ml of tetrahydrofuran were added.
100 ml of an aqueous M sodium hydroxide solution and 50 ml of a tetrahydrofuran solution of 4-fluorobenzenesulfonyl chloride (9.7 g) were simultaneously added dropwise. This solution was stirred at room temperature for 24 hours to react. After the reaction is completed, the reaction solution is adjusted to pH
Adjusted to 2-3 and extracted with ethyl acetate. The extract was washed with diluted hydrochloric acid and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a mixed solution of hexane-ethyl acetate to obtain 8.3 g of N- (4-fluorophenylsulfonyl) -L-valine as white crystals.

N- (4-fluorophenylsulfonyl)
-L-valine (8.0 g) and N-hydroxysuccinimide (4.4 g) are dissolved in 150 ml of tetrahydrofuran, and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride is stirred under ice-cooling. Salt (7.3
150 ml of a dichloromethane solution of g) were slowly added. The solution was stirred at room temperature for about 12 hours to react.
After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution, and N- (4-fluorophenylsulfonyl)
9.6 g of -D-valine N-hydroxysuccinimide ester was obtained as white crystals.

N- (4-fluorophenylsulfonyl)
-D-Valine N-hydroxysuccinimide ester (1.8 g) and D-leucinol (0.74 g) were added to 80 ml of dichloromethane, and triethylamine (1.5 g) was added with stirring at room temperature. The solution was stirred for 2 hours to react. After completion of the reaction, the solvent was distilled off under reduced pressure, the residue was dissolved in ethyl acetate, washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. Ethyl acetate was distilled off under reduced pressure, and the residue was washed with a hexane-ethyl acetate mixed solution to obtain 1.6 g of N- (4-fluorophenylsulfonyl) -D-valyl-D-leucinol as white crystals. N- (4-Fluorophenylsulfonyl) -D-valyl-D-leucinol (1.5 g) was dissolved in dimethylsulfoxide solution (20 ml) and dichloromethane (10 ml), and triethylamine (2.4 g) was added. Sulfur pyridine complex (2.6 g) dimethyl sulfoxide 20 m
was added, and the mixture was further stirred for 1 hour to react. After completion of the reaction, ethyl acetate was added, and the mixture was washed with dilute hydrochloric acid, a saturated aqueous solution of sodium hydrogen carbonate and saturated saline, and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the fractionated residue was purified using a TLC plate (developing solvent: hexane-ethyl acetate, 1: 1) to give N- (4-fluorophenylsulfonyl) -D-valyl-D-leucinal. (Compound 55)
1.0 g was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.74 (d, 3
H, J = 6.3Hz), 0.82 (d, 6H, J = 6.3Hz), 0.87 (d, 3H, J =
6.9Hz), 1.15-1.45 (m, 3H), 1.81-1.93 (m, 1H), 3.59
(t, 1H, J = 6.8Hz), 3.80-3.88 (m, 1H), 7.33-7.42 (m,
2H), 7.79-7.86 (m, 2H), 7.95 (d, 1H, J = 6.9Hz), 8.26
(d, 1H, J = 6.9Hz), 9.14 (s, 1H). Anal. (C ₂₂ H ₃₀ N ₂ O ₄ S) C, H, N.

Example 55-2 N- (4-Fluorophenylsulfonyl) -L-valyl-D-leucinal (N- (4-fluorophenylsulfonyl) -L-valyl-D-leucinal) was prepared in the same manner as in Example 55, except that valine was used in place of D-valine. Compound 56) was obtained as white crystals.

¹ H-NMR (DMSO-d ₆ 270 MHz) δ: 0.78 (d, 3
H, J = 6.3Hz), 0.82 (d, 3H, J = 6.6Hz), 0.83 (d, 6H, J =
6.3Hz), 1.18-1.50 (m, 3H), 1.79-1.92 (m, 1H), 3.61-
3.63 (m, 1H), 3.84-3.92 (m, 1H), 7.33-7.44 (m, 2H),
7.80-7.96 (m, 3H), 8.22 (d, 1H, J = 6.9Hz), 8.96
(s, 1H). Anal. (C ₂₀ H ₂₉ FN ₂ O ₄ S) C, H, N.

Example 56 (Tablet) Compound 35 30 mg Lactose 80 mg Starch 17 mg Magnesium stearate 3 mg A tablet is formed by a conventional method using the above ingredients as ingredients for one tablet. Sugar coating may be applied as necessary.

Example 57 (Injection) Compound 48 2.5 mg Sodium chloride 900 mg 1N Sodium hydroxide Appropriate amount Distilled water for injection Total amount 100 ml The above components are mixed by a conventional method to prepare an injection.

Example 58 (Eyedrops) Compound 35 50 mg Boric acid 700 mg Borax proper amount Sodium chloride 500 mg Hydroxymethylcellulose 500 mg Sodium edetate 0.05 mg Benzalkonium chloride 0.005 mg Total amount of sterilized purified water 100 ml Mix to form an ophthalmic suspension.

Test Example 1 Guinea Pig Angle by Basic FGF (bFGF) Implantation
Of cysteine protease inhibitors on membrane angiogenesis
Effect (Test method) (1) Method of preparing pellet for transplantation 5 μl of a dichloromethane solution of 8% ethylene vinyl acetate copolymer (EVA) was dropped on a Teflon plate and air-dried, and 3 μl of 0.0167 w / v% bFGF was dropped thereon and air-dried. Was. After drying, it is rolled and formed into a small rod shape and bF
A pellet containing GF (500 ng) was prepared. Similarly, a test drug-containing pellet was prepared by using a test solution instead of 0.0167 w / v% bFGF. The test drug-containing pellets were prepared to be 27-mer calpastatin peptide (manufactured by Sigma) 0.03 μmole, 0.1 μmole / pellet and leupeptin (manufactured by Peptide Institute) 0.1 μmole / pellet. As a control, a pellet containing only EVA was prepared and used as a base pellet. (2) Transplantation of pellet into guinea pig cornea M. Kusaka et al. [Biochem. Biophys. Res. Comm., 174
Volume, pp. 1070-1076 (1991)].
That is, a male guinea pig weighing 300 to 400 g is anesthetized with a 1: 1 mixture of Ketalal 50 (ketamine hydrochloride; Sankyo Co., Ltd.) and Ceractal (Xylazine hydrochloride; Bayer Yakuhin Co., Ltd.), and then both eyes Using a 0.5 mm wide ophthalmic spatula, a pocket was created from the limbus of the cornea toward the center of the cornea in the middle layer of the corneal stromal layer. The test drug-containing pellet was inserted into the created pocket, and the bFGF-containing pellet was inserted adjacent to the pellet. To prevent infection, ofloxacin ophthalmic ointment (Taribit ointment (ofloxacin 0.3%), Santen Pharmaceutical Co., Ltd.) was instilled once immediately after both pellets were inserted.
% Lomefloxacin (Lomeflon ophthalmic otic solution, manufactured by Senju Pharmaceutical Co., Ltd.) was instilled once a day for 5 days. (3) Determination of Effect of Cysteine Protease Inhibitor The effect of the cysteine protease inhibitor was determined by observation of the cornea using a slit lamp. In addition, blood vessels newly formed in the cornea, which is an avascular tissue, have high permeability, and it is presumed that the wet weight of the cornea and the plasma component increase due to the plasma component flowing into the cornea. They were sacrificed and the corneas were harvested. The collected cornea was measured for wet weight. After measuring the wet weight, homogenize the cornea,
Water-soluble proteins obtained by centrifugation were separated by SDS polyacrylamide gel electrophoresis. After electrophoresis, stain the gel with Coomassie Brilliant Blue,
Albumin, one of the main proteins of plasma components, was quantified by image analysis (NIH Image 1.31). Guinea pig serum albumin was used as a standard.

(Test Results) (1) Results of Corneal Observation Using Slit Lamp Compared with the untreated group of corneas,
No changes were observed in the cornea. In the bFGF-containing pellet transplantation group (hereinafter sometimes referred to as a control group),
Appearance of blood vessels was observed around the bFGF-containing pellet transplantation site 4 days after transplantation, and 9 days after transplantation, blood vessels were regenerated in the entire cornea of the guinea pig. In the group transplanted with the 27-mer calpastatin peptide-containing pellet and the leupeptin-containing pellet together with the bFGF-containing pellet, the appearance of blood vessels was observed in all groups, but the degree of the blood vessels was milder than that in the control group, and the blood vessels were mainly in the limbus of the cornea. Only a newborn was found. The state of the cornea of the guinea pig 9 days after the transplantation is shown in FIGS. 1 and 2. (2) Wet weight of cornea The results are shown in FIG. The corneal wet weight of the untreated group and the base pellet-transplanted group was almost the same, and no effect of the base pellet-transplant was observed. On the other hand, the corneal wet weight of the control group was increased as compared with the untreated group and the base pellet transplant group. In the group transplanted with the 27-mer calpastatin peptide-containing pellet or leupeptin-containing pellet together with the bFGF-containing pellet, the increase in corneal wet weight was suppressed as compared with the control group. From this, it was found that the 27-mer calpastatin peptide and leupeptin suppressed the increase in wet weight of the cornea due to angiogenesis. (3) Amount of albumin in cornea The results are shown in FIG. The untreated group had an albumin level of 3
5.2 ± 9.6 (SD) or 56.1 ± 13.5
(SD). This is because the cornea is avascular tissue. In comparison, the amount of albumin in the group implanted with the base pellet was slightly higher than that in the untreated group. This was thought to be due to surgical stimulation during pellet implantation. In contrast, the amount of albumin in the control group increased about 17-fold or about 9-fold in the untreated group, and about 8-fold in the base pellet-transplanted group. This is because newly formed blood vessels have high permeability, and albumin which is a main component of plasma is increased corresponding to the newly formed blood vessels. In the group transplanted with the 27-mer calpastatin peptide-containing pellet and the leupeptin-containing pellet together with the bFGF-containing pellet, the increase in the amount of albumin in the cornea was suppressed as compared with the control group. As described above, it has been clarified that the cysteine protease inhibitor suppresses angiogenesis.

Test Example 2 The biological activity of the compounds represented by formulas (I) and (IV) is described. The compounds (I), (IV) and salts thereof exhibit a thiol protease inhibitory activity. The following methods were used to measure the inhibitory activities of calpain, cathepsin L and papain, and serine protease on trypsin, and to determine the results. The results are shown in Tables 6 and 7. μ-calpain inhibitory activity The activity of μ-calpain (manufactured by Nacalai Tesque, Inc.)
nal. Biochem., 208, 387-392 (1993)]. That is, 0.5 mg /
ml casein, 50 mM Tris-HCl (pH 7.4), 20
To a solution containing mM dithiothreitol and 4 mM calcium chloride, 2.5 μl of a dimethyl sulfoxide solution containing various concentrations of a test drug and 0.03 enzyme unit μ-calpain were added to initiate a reaction. The final liquid volume was 250 μl.
After reacting at 30 ° C. for 60 minutes, 100 μl of the reaction solution was transferred to another container, 50 μl of purified water and 100 μl of 50% Coomassie brilliant blue solution were added, the mixture was allowed to stand at room temperature for 15 minutes, and the absorbance at 595 nm was measured. . After adding 2.5 μl of a dimethyl sulfoxide solution containing no test drug and treating in the same manner, the measured value was used as a control value, 0.2 mM instead of 4 mM calcium chloride aqueous solution.
Using EDTA as a blank value, the inhibition rate was calculated by the following formula, and the relationship with the concentration of the inhibitor was plotted on a logarithmic graph, and the amount required for 50% inhibition (IC ₅₀ )
I asked.

[0281]

(Equation 1)

Assay for Cathepsin L Inhibitory Activity The activity of cathepsin L (manufactured by Cosmo Bio), which is a cysteine protease, is described in the literature [Methods in Enzymology, 80].
Vol., 535-561 (1981)]. That is, 85 mM acetate buffer (pH 5.5), 2
mM dithiothreitol, 1 mM EDTA, 2 μg cathepsin L, 20 μM
Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-
50 μl of Coumaryl-7-Amide (Z-Phe-Arg-MCA) was added,
The reaction was started with a final volume of 200 μl. 30 ° C,
After reacting for 20 minutes, 1M Tris-HCl (pH 8.0)
Was added to stop the reaction. The amount of released 4-methyl-7-aminocoumarin was measured using a fluorometer at an excitation wavelength of 360 nm and a fluorescence wavelength of 450 nm. Control values were measured after processing in the same manner under test without addition, those enzymes not added as a blank value, was calculated and IC ₅₀ in the same manner as described above.

Measurement of papain and trypsin inhibitory activity
Methods The activities of papain, a cysteine protease, and trypsin, a serine protease (manufactured by Sigma), were measured according to the method described in the literature [Anal. Biochem., 208, 387-392 (1993)]. That is, 0.5 mg / m
l casein, 50 mM Tris-HCl (pH 8.
0), 20 mM dithiothreitol, 0.2 mM ED
2.5 μl of a dimethyl sulfoxide solution containing various concentrations of the test drug and 0.03 enzyme units of papain or trypsin were added to the solution containing TA, and the reaction was started. The final liquid volume was 250 μl. After reacting at 30 ° C. for 60 minutes, 100 μl of the reaction solution was transferred to another container, and 50 μl of purified water was added.
l and 100 μl of 50% Coomassie Brilliant Blue solution
Was added and left at room temperature for 15 minutes, and then the absorbance at 595 nm was measured. It was added dimethyl sulfoxide solution 2.5μl containing no test drug, similarly treated control values were measured after, those enzymes not added as a blank value, was calculated and IC ₅₀ in the same manner as described above.

[0284]

[Table 7]

[0285]

[Table 8]

The cysteine protease inhibitory compound used in the present invention did not show toxicity to humans and animals.

[0287]

Industrial Applicability The angiogenesis inhibitor of the present invention suppresses the formation of blood vessels in living tissues, so that angiogenesis accompanying wound healing, inflammation, tumor growth, diabetic retinopathy, retina of premature infants It can be used as an excellent therapeutic and prophylactic agent for angiogenesis seen in diseases such as retinal vein occlusion and senile discoid macular degeneration, and as a preventive agent for tumor metastasis. Thus, the compounds represented by the general formulas (I) and (IV) and salts thereof have an inhibitory activity on cysteine proteases such as calpain, cathepsin L and papain, and have an inhibitory activity on serine protease (trypsin). I didn't. Therefore, Compounds (I), (IV) and salts thereof are useful for various diseases involving cysteine protease, for example, mammals (eg, mice, rats, rabbits, dogs, cats, cows, pigs, humans, etc.). It is useful as a prophylactic and therapeutic agent for ischemic disease, inflammation, muscular dystrophy, immune disease, essential hypertension, Alzheimer's disease, subarachnoid hemorrhage and osteoporosis, and particularly as an anti-angiogenic agent.

[Brief description of the drawings]

FIG. 1: bFGF-containing pellet and 27-mer calpastatin peptide (0.03 μmole and 0.1 μmole)
le) A drawing (a photograph showing the form of an organism) showing the state of the guinea pig cornea observed by a slit lamp after transplanting the bFGF-containing pellet together with the bFGF-containing pellet into the guinea pig cornea, 9 days later.

FIG. 2 is a drawing showing the state of the guinea pig cornea observed by a slit lamp 9 days after transplanting the bFGF-containing pellet and the leupeptin (0.1 μmole) -containing pellet together with the bFGF-containing pellet into a guinea pig cornea. Photo).

FIG. 3. bFGF-containing pellet and 27-mer calpastatin peptide (0.03 μmole and 0.1 μmole)
FIG. 11 is a graph showing the wet weight of guinea pig cornea collected after transplanting a pellet containing le) or leupeptin (0.1 μmole) into a guinea pig cornea together with a pellet containing bFGF.

FIG. 4. bFGF-containing pellet and 27-mer calpastatin peptide (0.03 μmole and 0.1 μmole)
le) containing pellet or leupeptin (0.1μmole)
It is a graph which transplanted the containing pellet to the guinea pig cornea with the bFGF containing pellet, and collect | recovered 9 days afterward and is a graph which shows the amount of albumin in the guinea pig cornea.

──────────────────────────────────────────────────の Continuation of the front page (51) Int.Cl. ⁶ Identification code FI A61K 38/55 ABN A61K 45/00 AED 45/00 AED C07D 209/14 C07D 209/14 303/48 303/48 405/06 207 405/06 207 C07K 14/81 // C07K 14/81 A61K 37/64 ABN (C07D 405/06 207: 16 303: 48) (72) Inventor Hiroka Yoshida 634-141 Wada-cho, Nishiwaki-shi, Hyogo Prefecture

Claims (33)

[Claims] 1. An angiogenesis inhibitor comprising a cysteine protease inhibitory compound. 2. A cysteine protease inhibitor compound,
The angiogenesis inhibitor according to claim 1, which is a calpain inhibitory compound. 3. The angiogenesis inhibitor according to claim 2, wherein the calpain inhibitory compound is one or more compounds selected from calpastatin and calpastatin peptide. 4. The calpastatin peptide has the following general formula: -Gly-A-Tyr-Arg- (where A is -Lys-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys
-, -Lys-Arg-Glu-Val-Thr-Leu-Pro-Pro-Lys-, -Glu-As
p-Asp-Glu-Thr-Ile-Pro-Ser-Glu-, -Glu-Asp-Asp-Glu-
Thr-Val-Pro-Pro-Glu-, -Glu-Asp-Asp-Glu-Thr-Val-Pr
o-Ala-Glu-, -Glu-Lys-Glu-Glu-Thr-Ile-Pro-Pro-Asp-
Or -Glu-Arg-Asp-Asp-Thr-Ile-Pro-Pro-Glu-. 4. The angiogenesis inhibitor according to claim 3, which is one or more compounds selected from peptides having the amino acid sequence of (3). 5. The angiogenesis inhibitor according to claim 4, wherein the calpastatin peptide has the following amino acid sequence. Asp-Pro-Met-Ser-Ser-Thr-Tyr-Ile-Glu-Glu-Leu-Gly-Ly
s-Arg-Glu-Val-Thr-Ile-Pro-Pro-Lys-Tyr-Arg-Glu-Leu-
Leu-Ala 6. The calpain inhibitory compound is one or more compounds selected from substances that inhibit a Ca ²⁺ binding site having high homology to calmodulin of calpain.
An angiogenesis inhibitor according to the above. 7. The angiogenesis inhibitor according to claim 6, wherein the substance that inhibits a Ca ²⁺ binding site having a high homology to calmodulin of calpain is at least one compound selected from calmodulin antagonistic compounds. 8. The cysteine protease inhibitor compound,
2. The compound according to claim 1, which is at least one compound selected from an epoxysuccinic acid peptide compound, a peptide aldehyde compound, a peptide halomethane compound, a peptide diazomethane compound, a peptide halohydrazide compound, a peptide disulfide compound, a peptide ketoamide compound and an isocoumarin compound. Angiogenesis inhibitors. 9. The cysteine protease inhibitor compound,
The angiogenesis inhibitor according to claim 8, which is an epoxysuccinic acid peptide compound. 10. The epoxy succinate peptide compound,
Formula (I): [Wherein, R ¹ represents a carboxyl group which may be esterified or a carboxamide which may be substituted, and R ² represents hydrogen or a lower alkyl group, or a ring linked to R ³ or R ⁴ May form R ³
And R ⁴ are the same or different and represent hydrogen, an optionally substituted lower alkyl group or an optionally substituted sulfide group, and R ³ and R ⁴ may combine to form a ring; ⁵ is of the formula (II) (Wherein R ⁶ represents a halogen atom or an alkoxy group), or a formula (III) SO ₂ —R ⁷ (III) wherein R ⁷ is substituted with a lower alkyl group. An aryl group or an amino group which may be substituted), and n represents 0 or 1.] or a salt thereof. Agent. 11. The angiogenesis inhibitor according to claim 10, wherein R ¹ is a carboxyl group which may be esterified, or a carboxamide group which may be substituted with a hydroxy group or an aralkyloxy group. 12. The angiogenesis inhibitor according to claim 10, wherein R ² is hydrogen or a methyl group. 13. The angiogenesis inhibitor according to claim 10, wherein R ² is linked to R ³ or R ⁴ to form a pyrrolidine ring. 14. The method according to claim 10, wherein R ³ and R ⁴ are the same or different and each is a sulfide group optionally substituted with hydrogen, an aromatic or a lower alkyl group optionally substituted with a carbamoyl group or an acylamino group. Angiogenesis inhibitors. 15. The angiogenesis inhibitor according to claim 10, wherein R ³ and R ⁴ are linked to form a cyclopentane ring. 16. The angiogenesis inhibitor according to claim 10, wherein R ^{6 in the} formula (II) is chlorine or fluorine. 17. The angiogenesis inhibitor according to claim 10, wherein R ^{7 in the} formula (III) is a phenyl group or a dimethylamino group which may be substituted with a lower alkyl group. 18. The angiogenesis inhibitor according to claim 8, wherein the cysteine protease inhibitory compound is a peptide aldehyde compound. 19. The angiogenesis inhibitor according to claim 18, wherein the peptide aldehyde compound is leupeptin. 20. The peptide aldehyde compound represented by the formula (V)
I): embedded image (Wherein, R ¹¹ represents an aryl group having 6 to 10 carbon atoms which may have a substituent, and R ¹² and R ¹³ are the same or different,
It may represent hydrogen or an alkyl group having 1 to 4 carbon atoms or may be connected to form a ring having 3 to 7 carbon atoms, and R ¹⁴ is substituted with an aryl group, a cycloalkyl group or an aromatic heterocyclic ring. 20) a compound represented by the formula (1) or a salt thereof. 21. The angiogenesis inhibitor according to claim 20, wherein R ¹¹ is a phenyl group or a naphthyl group which may be substituted with a fluorine, chlorine or methyl group. 22. R ¹¹ is 4-fluorophenyl, 4-
The angiogenesis inhibitor according to claim 21, which is a group selected from chlorophenyl, p-tolyl and 2-naphthyl. 23. R ¹² is propyl, an isopropyl or tert- butyl, claim 2 R ¹³ is hydrogen
0. The angiogenesis inhibitor according to 0. 24. The angiogenesis inhibitor according to claim 23, wherein R ¹² is isopropyl and R ¹³ is hydrogen. 25. The angiogenesis inhibitor according to claim 20, wherein the ring formed by linking R ¹² and R ¹³ is cyclohexylidene. 26. The angiogenesis inhibitor according to claim 20, wherein R ¹⁴ is isobutyl, benzyl, cyclohexylmethyl or indol-3-ylmethyl. 27. Formula (VI): (Wherein, R ¹¹ represents an aryl group having 6 to 10 carbon atoms which may have a substituent, and R ¹² and R ¹³ are the same or different,
It may represent hydrogen or an alkyl group having 1 to 4 carbon atoms or may be connected to form a ring having 3 to 7 carbon atoms, and R ¹⁴ is substituted with an aryl group, a cycloalkyl group or an aromatic heterocyclic ring. A lower alkyl group which may be substituted) or a salt thereof. 28. The compound or a salt thereof according to claim 27, wherein R ¹¹ is a phenyl group or a natyl group optionally substituted with fluorine, chlorine or methyl group. 29. R ¹¹ is 4-fluorophenyl, 4-
The compound according to claim 27, which is a group selected from chlorophenyl, p-tolyl and 2-naphthyl, or a salt thereof. 30. R ¹² is propyl, an isopropyl or tert- butyl, claim 2 R ¹³ is hydrogen
7. The compound according to 7, or a salt thereof. 31. The compound according to claim 27, wherein R ¹² is isopropyl and R ¹³ is hydrogen, or a salt thereof. 32. The compound or a salt thereof according to claim 27, wherein the ring formed by linking R ¹² and R ¹³ is cyclohexylidene. 33. The compound according to claim 27, wherein R ¹⁴ is isobutyl, benzyl, cyclohexylmethyl or indol-3-ylmethyl, or a salt thereof.