JPH07157472A - 2-piperazinone-1-acetic acid derivative and its use - Google Patents

Abstract

PURPOSE:To obtain the subject new compound useful as an antithrombotic agent due to its manifestation of stronger inhibiting actions on cell adhesion in a small dose, excellence in persistence thereof and suppressing actions on blood platelet agglutination with adenosine diphosphate(ADP) for a long period by the oral administration. CONSTITUTION:This compound is expressed by formula I [A is amidino or aminoethyl; R<10> is nitro, a halogen, etc.; R<12> and R<13> are each H, OH, etc.; X is OH, p-hydroxyphenyl, etc.; Y is carboxyl; (n) is 1 or 2] or its salt, e.g. (S)-4-(4-amidinobenzoyl-L-tyrosyl)-3-methoxycarbonylmethyl-2-oxopipera zine-1- acetic acid. The compound is obtained by condensing, e.g. a compound expressed by formula II with a compound expressed by formula III in a solvent such as THF in the presence of a base using a peptide-forming reagent such as dicyclohexylcarbodiimide at 0 deg.C to ambient temperature.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel 2-piperazinone-1-acetic acid derivative having a cell adhesion inhibitory action or a salt thereof and an animal cell adhesion inhibitor containing them as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION The present invention relates generally to the provision of therapeutic agents for various diseases by regulating or inhibiting cell adhesion. Fibronectin, vitronectin, osteopontin, collagen, thrombospondin, fibrinogen, and von Willebrand factor are known as factors involved in the adhesion of animal cells to the extracellular matrix. These proteins are tripeptide-
It contains Arg-Gly-Asp- as a cell recognition site. This tripeptide is a heterodimeric protein consisting of two membrane-bound subunits, the receptor integrin (in
is recognized by at least one protein belonging to tegrin (E. Ruoslahti and MD Pierschbacher,
Science, 238 , 491 (1987)). A structurally related integrin receptor that recognizes the amino acid sequence -Arg-Gly-Asp- is known to be expressed on the extracellular surface of platelets, endothelial cells, leukocytes, lymphocytes, monocytes, and granulocytes. ing. The compound having the amino acid sequence -Arg-Gly-Asp- inhibits the binding of the cell adhesion factor by competitively binding with the adhered site to which the cell adhesion factor binds. Examples of such cell adhesion inhibitors include H
-Gly-Arg-Gly-Asp-Ser-Pro-OH is known. When a blood vessel is damaged, platelets are activated by collagen or the like under the blood vessel, fibrinogen binding to the platelet, that is, platelet aggregation occurs, and a thrombus is formed.
The interaction between platelets and fibrinogen is GP IIb / II
It occurs via Ia, which is an important feature of platelet aggregation. Cell adhesion inhibitors are thrombin, epinephrine, A
It is possible to inhibit platelet aggregation due to platelet aggregation-inducing substances such as DP and collagen. Further, a cell adhesion inhibitor is expected as a drug for suppressing the metastasis of cancer cells (preventing adhesion and immobilization at the destination of metastasis). As a conventionally known cell adhesion inhibitor, a linear peptide or cyclic peptide containing an amino acid sequence of -Arg-Gly-Asp- (RGD) is known. Examples of them include the Journal of Biological Chemistry (J. Biol. Chem.), 262 , 172.
94 (1987) and JP-A-2-174797.

The above-mentioned known peptide derivatives cannot be said to have sufficient activity, and their oral absorbability is not satisfactory. In addition, these peptide derivatives are hydrolyzed by enzymes such as aminopeptidases, carboxypeptidases, and various endopeptidases such as serine proteases, and their stability in a solution in which these enzymes are present or in vivo is insufficient, and thus clinical application as a drug. In order to do so, it has not been fully resolved. Further, non-peptide compounds having an antithrombotic effect are described in JP-A-4-264068 and European Patent Application Publication No. 505868. A compound having a 4- to 7-membered cyclic alkyleneimino such as a pyrrolidine ring and a compound having a piperidine ring are described. Furthermore, a compound having a piperidinone ring having an inhibitory effect on cell adhesion has been published in European patent application.
No. 529858 is disclosed. There is a need for compounds having stronger and more persistent activity than these known compounds having antithrombotic activity.

[0004]

DISCLOSURE OF THE INVENTION The object of the present invention is to provide a compound having a stronger and longer-lasting cell adhesion-inhibiting action at a low dose, as compared with the conventionally known compounds having a cell-adhesion inhibiting action. That is. That is, the present invention relates to a novel 2-piperazinone-1-acetic acid derivative which has solved the above-mentioned problems, and a drug containing the same as an active ingredient and having an action of inhibiting cell adhesion. That is, the present invention has the formula (I)

[Chemical 3] [In the formula, A represents an amidino group or an optionally substituted aminoethyl group, and R ¹⁰ represents a nitro group, a halogen atom, a lower alkenyl group, a lower alkynyl group, a hydroxymethyl group, a formyl group, a lower alkyloxycarbonyl group. And a group represented by the formula —OR ¹¹ (in the formula, R ¹¹ represents a hydrogen atom or an optionally substituted lower alkyl, lower alkenyl, lower alkynyl, lower alkanoyl, carbamoyl or methanesulfonyl group). R ¹² and R ¹³ each represent a hydrogen atom, a hydroxyl group, a lower alkoxy group or a halogen atom, X represents a hydroxyl group,
A p-hydroxyphenyl group or a carboxyl group which may be esterified or amidated is shown, Y is a carboxyl group which may be esterified or amidated, and n is 1 or 2. ] It is related with the compound represented by these, or its salt, and the cell adhesion inhibitor containing these compounds. A typical compound represented by formula (I) is formula (I ′)

[0005]

[Chemical 4] [In the formula, R ¹¹ represents a hydrogen atom, an optionally substituted lower alkyl group, a lower alkanoyl group, an optionally substituted carbamoyl group or a methanesulfonyl group, and R ¹² and R ¹³ represent a hydrogen atom and a hydroxyl group, respectively. Represents a lower alkoxy group or a halogen atom, X represents a hydroxyl group, p-hydroxyphenyl group or a carboxyl group which may be esterified or amidated, and Y represents a carboxyl group which may be esterified or amidated. Shown, n is 1
Or 2] or a salt thereof. A represents an amidino group or an optionally substituted aminoethyl group. The substituted aminoethyl group is preferably a group that can be converted into a physiologically active free aminoethyl group by elimination of the substituent in vivo after administration as a prodrug. Examples of the amino substituent of such a substituted aminoethyl group include pivaloyloxymethyl, n-octyloxycarbonyl, n-hexyloxycarbonyl, n-octylaminocarbonyl, n-hexylaminocarbonyl and tetrahydrofuran-2-yl. , Pyrrolidin-1-ylmethyl, morpholinomethyl, N, N-dimethylaminomethylaminocarbonyloxymethyl group and the like. As the group or atom represented by R ¹⁰ ,
For example, a nitro group, a halogen atom, a lower alkenyl group, an alkynyl group, a hydroxymethyl group, a formyl group, a lower alkyloxycarbonyl group and a group bonded through O represented by the formula —OR ¹¹ are preferable. R ¹¹ is a hydrogen atom or optionally substituted lower alkyl,
Lower alkenyl, lower alkynyl, lower alkanoyl,
A carbamoyl group, a methanesulfonyl group and the like are preferable.

That is, R ¹⁰ is a nitro group, a halogen atom, a lower alkenyl group, a lower alkynyl group, a hydroxymethyl group, a formyl group, a lower alkyloxycarbonyl group and a formula OR ¹¹ (wherein R ¹¹ is a hydrogen atom or a hydrogen atom, respectively). (Indicated is optionally substituted lower alkyl, lower alkenyl, lower alkynyl, lower alkanoyl optionally substituted carbamoyl or methanesulfonyl group.)
It is preferably one selected from the group represented by
More specifically, R ¹⁰ is a hydroxyl group, a C _1-5 alkoxy group which may be substituted with a C _1-3 alkoxy group, a C _2-3
Alkenyloxy group, C _2-3 alkynyloxy group, nitro group, halogen atom, C _1-3 alkanoyloxy group, C
Carbamoyloxy group optionally substituted with _1-3 alkyloxy group, methanesulfonyloxy group, C _2-3 alkenyl group, C _2-3 alkynyl group and hydroxymethyl group, formyl group, C _1-3 alkyloxycarbonyl One selected from the group is preferable. Examples of the lower alkyl group represented by R ¹¹ include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl and tert.
A linear or branched alkyl group having 1 to 5 carbon atoms such as -butyl, n-pentyl, i-pentyl is preferable. Particularly, a straight-chain C _1-3 alkyl group is preferable.

Substituents of the optionally substituted lower alkyl, lower alkenyl, lower alkynyl, lower alkanoyl and methanesulfonyl groups represented by R ¹¹ include, for example, hydroxyl group, lower (C _1-3 ) alkoxy group, amino Base,
Mono-lower (C _1-3 ) alkylamino group, di-lower (C _1-3 ).
Alkylamino group, lower (C _1-3 ) alkanoyl group, lower (C _1-3 ) alkanoyloxy group, lower (C _1-3 ) alkanoylamino group, lower (C _1-3 ) alkoxycarbonyl group, lower (C _1-3 ) Examples thereof include a carbamoyl group which may be substituted with an alkyl group. Among these substituents, lower (C _1-3 ) alkoxy groups such as methoxy, ethoxy and propyloxy are particularly preferable. Further, examples of the substituent of the optionally substituted carbamoyl group represented by R ¹¹ include lower (C _1-3 ) alkyl. Lower alkoxy group represented by R ¹² and R ¹³ and optionally substituted lower alkyl, lower alkenyl, lower alkynyl, lower alkanoyl represented by R ¹¹ and lower alkoxy contained in the substituent of methanesulfonyl group. As the group, an alkoxy group having 1 to 3 carbon atoms such as methoxy, ethoxy and n-propyloxy is preferable. A lower alkanoyl group represented by R ¹¹ and an optionally substituted lower alkyl, lower alkenyl, lower alkynyl represented by R ¹¹ .
The lower alkanoyl group contained in the substituents of the lower alkanoyl and methanesulfonyl groups is preferably an alkanoyl group having 1 to 3 carbon atoms such as formyl, acetyl and propionyl. Examples of the halogen atom represented by R ¹⁰ , R ¹² and R ¹³ include fluorine, chlorine, bromine and iodine. Especially, a fluorine atom and a chlorine atom are preferable. Lower alkyl group of the substituents of the optionally substituted carbamoyl group represented by R ^11, which may lower each be substituted a lower alkyl group and R ¹¹ contained in lower alkyloxycarbonyl groups represented by R ¹⁰ Substituted amino groups as substituents of alkyl, lower alkenyl, lower alkynyl, lower alkanoyl and methanesulfonyl groups and lower alkyl groups as substituents of substituted carbamoyl groups are carbon such as methyl, ethyl, n-propyl, i-propyl and the like. A lower alkyl group having a number of 1 to 3 is preferable.

The lower alkenyl group represented by R ¹⁰ and R ¹¹ is preferably an alkenyl group having 2 to 3 carbon atoms such as vinyl, allyl and 1-propenyl. The lower alkynyl group represented by R ¹⁰ and R ¹¹ is preferably an alkynyl group having 2 to 3 carbon atoms such as ethynyl and propargyl. Among the above compounds, the compound in which R ¹⁰ is a hydroxyl group or a methoxy group is most preferable. Also preferred are compounds of the formula (I) in which R ¹⁰ is an ethoxy group, a methoxyethoxy group, a propoxy group, a propargyloxy group, an allyloxy group, a fluorine atom or a chlorine atom, and among these, an ethoxy group, a propargyloxy group, An allyloxy group and a fluorine atom are preferred. A compound having a hydrogen atom is particularly preferable for both R ¹² and R ¹³ . Also, regarding n, n
Compounds with = 1 are preferred. A is an amidino group,
A compound in which R ¹⁰ is a hydroxyl group or a methoxy group, R ¹² and R ¹³ are both hydrogen atoms, and n = 1 is particularly preferable. A compound in which A is an amidino group, R ¹⁰ is an ethoxy group, a propargyloxy group, an allyloxy group or a fluorine atom, R ¹² and R ¹³ are both hydrogen atoms, and n = 1 is also preferable. A is an aminoethyl group and R ¹⁰
Is also a hydroxyl group or a methoxy group, R ¹² and R ¹³ are both hydrogen atoms, and a compound in which n = 1 is also preferable. Further, a compound in which A is an aminoethyl group, R ¹⁰ is an ethoxy group, a propargyloxy group, an allyloxy group or a fluorine atom, R ¹² and R ¹³ are both hydrogen atoms, and n = 1 is also preferable.

The carboxyl group which may be esterified or amidated represented by X and Y is a group which can be converted into a physiologically active compound in vivo by administering Compound (I) as a prodrug. But X and Y are each of the formula

[Chemical 5] Can be expressed as R ¹ and R ² are usually the same or different and each represent a hydroxyl group, an alkoxy group having 1 to 8 carbon atoms (eg, methoxy, ethoxy, propoxy, butoxy),
Examples of the lower alkenyloxy group include alkenyloxy groups having 3 to 12 carbon atoms such as allyloxy and butenyloxy, and aralkyloxy groups (for example, lower alkyl moieties such as benzyloxy, phenethyloxy, and 3-phenylpropyloxy have 1-4 carbon atoms). Phenyl lower alkyloxy groups) or each -NR
³ R ⁴ represents an optionally substituted amino group represented by —NR ⁵ R ⁶ . In NR ³ R ⁴ and NR ⁵ R ⁶ , R ³ and R ⁴ , R ⁵ and R ⁶ are the same or different and each is hydrogen,
Lower alkyl group (methyl, ethyl, propyl, butyl,
A lower alkyl group having 1 to 6 carbon atoms such as hexyl), an alkenyl group having 3 to 8 carbon atoms (eg, allyl, 2-
Butenyl, 3-pentenyl), or an aralkyl group having 6 to 12 carbon atoms (eg, benzyl, phenethyl, phenylpropyl, pyridylmethyl), wherein the aryl group in the aralkyl group is unsubstituted or 1 to 2 May be substituted with one substituent. The substituent is nitro,
Halogen (chlorine, fluorine, bromine), lower alkyl group (methyl, ethyl, propyl), lower alkoxy group (methoxy, ethoxy, propoxy) and the like can be mentioned.

When the compound (I) of the present invention is used as a prodrug type oral preparation, R ¹ and R ² are hydroxyl groups or optionally substituted amino [(eg, amino, N
-Lower (C _1-4 ) alkylamino, N, N-dilower (C
_1-4 ) Alkylamino, etc.)] or an optionally substituted alkoxy group [eg, an alkyl part is a hydroxyl group, optionally substituted amino (eg, amino, dimethylamino,
Diethylamino, piperidino, morpholino, etc.), halogen, lower (C _1-6 ) alkoxy, lower (C _1-6 ) alkylthio, lower (C _1-6 ) alkyloxycarbonyl (eg, butyloxycarbonyl), propylidene, 3-
Phthalidylidene, optionally substituted aminocarbonyl or optionally substituted dioxolenyl (eg,
5-methyl-2-oxo-1,3-dioxolen-4-
Lower (C _1-6 ) alkoxy group which may be substituted with, for example, formula —OCH (R ⁷ ) OCOR ⁸ [wherein R ⁷
Is hydrogen or a linear or branched lower alkyl group having 1 to 6 carbon atoms (eg, methyl, ethyl, n-propyl, ylpropyl, n-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, neopentyl. Etc.) or a cycloalkyl group having 5 to 7 carbon atoms (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.), and R ⁸ is a linear or branched lower alkyl group having 1 to 6 carbon atoms (eg, methyl). , Ethyl, n-propyl, isopropyl,
n-butyl, isobutyl, sec-butyl, t-butyl,
n-pentyl, isopentyl, neopentyl, etc.), lower alkenyl group having 2-8 carbon atoms (eg, vinyl, propenyl, allyl, isopropenyl, etc.), cycloalkyl group having 5-7 carbon atoms (eg, cyclopentyl, cyclohexyl, cyclo) Heptyl), a cycloalkyl group having 5 to 7 carbon atoms (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.) or a lower alkyl group having 1 to 3 carbon atoms substituted with an aryl group such as phenyl group (eg, benzyl, p -Chlorobenzyl, phenethyl, cyclopentylmethyl, cyclohexylmethyl, etc.), a cycloalkyl group having 5 to 7 carbon atoms (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.), or an aryl group such as a phenyl group having 2-3 carbon atoms. Lower alkenyl groups (eg, cinnamyl, etc.) Vinyl, propenyl, allyl, such as those having alkenyl portions of such isopropenyl), an aryl group such as phenyl group which may be substituted (e.g.,
Phenyl, p-tolyl, naphthyl, etc.), carbon number 1-6
A linear or branched lower alkoxy group (eg, methoxy, ethoxy, n-propoxy, isopropoxy, n-
Butoxy, isobutoxy, sec-butoxy, t-butoxy, n-pentyloxy, isopentyloxy, neopentyloxy, etc.), straight-chain or branched lower alkenyloxy group having 2-8 carbon atoms (eg, aryloxy, isobuteroxy, etc.) ), A cycloalkyloxy group having 5 to 7 carbon atoms (eg, cyclopentyloxy, cyclohexyloxy,
Cycloheptyloxy, etc.), a C 5-7 cycloalkyl group (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.) or an optionally substituted aryl group such as phenyl, etc. Groups (eg, those having an alkoxy moiety such as benzyl, phenethyloxy, cyclopentylmethyloxy, cyclohexylmethyloxy, methoxy, ethoxy, n-propoxy, isopropoxy, etc.), 5 to 7 carbon atoms
C2-substituted with a cycloalkyl group (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.) or an optionally substituted aryl group such as phenyl.
3 lower alkenyloxy groups (eg, those having an alkenyloxy moiety such as vinyloxy such as cinnamyloxy, propenyloxy, allyloxy, isopropenyloxy, etc.), aryloxy groups such as an optionally substituted phenoxy group (eg, phenoxy, p- Group, which represents nitrophenoxy, naphthoxy, etc.) is preferably introduced.

Particularly, preferred groups of the esterified carboxyl group as the esterified carboxyl group represented by Y when used as a prodrug are -COOMe, -COOEt, -COOtBu, -COO.
Pr, pivaloylmethoxycarbonyl, 1- (cyclohexyloxycarbonyloxy) ethoxycarbonyl, 5
-Methyl-2-oxo-1,3-dioxolen-4-ylmethoxycarbonyl, acetoxymethyloxycarbonyl, propionyloxymethoxycarbonyl, n-butyryloxymethoxycarbonyl, isobutyryloxymethoxycarbonyl, 1- (ethoxycarbonyloxy) ) Ethoxycarbonyl, 1- (acetyloxy) ethoxycarbonyl, 1- (isobutyryloxy) ethoxycarbonyl, cyclohexylcarbonyloxymethoxycarbonyl, benzoyloxymethoxycarbonyl, cinnamyloxycarbonyl, cyclopentylcarbonyloxymethoxycarbonyl, N, N-dimethylamino Carbonyl methoxy, 2
Examples include-(isobutyryloxycarbonyl) -2-propylideneethoxycarbonyl and (3-phthalidylidene) ethoxycarbonyl. In the formula (I), X is especially —COOH, —COOCH ₃ , or —CON.
H _{2 and the} like are preferable. Further, Y is preferably —COOH or a group capable of being converted into a carboxyl group in vivo. In the formula (I), X is —COOCH ₃ , Y is —COOH or a group capable of being converted into a carboxyl group in vivo, and A,
The compounds in which R ¹⁰ , R ¹² , R ¹³ and n are the above-mentioned preferred groups are particularly excellent in activity.

The compound (I) of the present invention has one or more asymmetric carbons in the molecule, and both the R configuration and the S configuration are included in the present invention with respect to these asymmetric carbons. The compound (I) may be a hydrate, and the hydrate and the hydrate are hereinafter referred to as the compound (I). Examples of the salt of the compound (I) include inorganic acid salts such as hydrochloride, hydrobromide, sulfate, nitrate and phosphate, for example, acetate, tartrate, citrate, fumarate and maleate. , Organic salts such as toluene sulfonate and methane sulfonate, metal salts such as sodium salt, potassium salt, calcium salt, aluminum salt, such as triethylamine salt, guanidine salt,
Examples thereof include pharmacologically acceptable salts such as salts with bases such as ammonium salts, hydrazine salts, quinine salts and cinchonine salts.

Specific preferred compounds include:
(S) -4- (4-Amidinobenzoyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid, (S) -4- (4-amidinobenzoyl-L-tyrosyl) -3 -Methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid hydrochloride, (S)-
4- (4-amidinobenzoyl-L-tyrosyl) -3-
Methoxycarbonylmethyl-2-oxopiperazine-1
-Acetic acid methanesulfonate, (S) -4- (4-amidinobenzoyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-
Acetic acid, (S) -4- (4-amidinobenzoyl-O-ethyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid, (S) -4- (4
-Aminoethylbenzoyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid and the like.

The compound (I) of the present invention [including the compound of the formula (I ')] can be produced, for example, by the following method. In the following description of the production method, it is included in the raw material compounds and intermediates. R ¹⁰ , R ¹² and R ¹³ may have a protecting group usually used in the peptide field, and the following description shows that these protected groups are also included.
Needless to say, the introduction and elimination of these functional groups can be carried out by conventional techniques. a) expression

[Chemical 6] [Wherein each of the symbols has the same meaning as defined above] is subjected to a condensation reaction, or b)

[Chemical 7] [Wherein each symbol has the same meaning as defined above] and a compound represented by the formula

[Chemical 8] It can be produced by subjecting a compound represented by the formula [each symbol is as defined above] to a condensation reaction.

The condensation reaction of the method a) and the method b) as a method for producing the compound (I) of the present invention includes an amide bond forming reaction in a conventional peptide, for example, active ester or mixed acid anhydride or acid chloride. It can be carried out by the method of. For example, compound (II) and compound (II
In the condensation reaction of I) or compound (IV) with compound (V), compound (II) or compound (IV) and 2,4,5-trichlorophenol, pentachlorophenol, 2-nitrophenol, or 4-nitrophenol , N-hydroxysuccinimide, N-
N-hydroxy compounds such as hydroxy-5-norbornene-endo-2,3-dicarboximide (HONB), 1-hydroxybenztriazole (HOBT) and N-hydroxypiperidine are condensed in the presence of a catalyst such as dicyclohexylcarbodiimide. After conversion to the active ester form, the condensation can be performed. It is also possible to obtain a mixed acid anhydride by reacting the compound (II) or the compound (IV) with isobutyl chloroformate, followed by condensation.

The condensation reaction between the compound (II) and the compound (III) or the compound (IV) and the compound (V) is carried out by dicyclohexylcarbodiimide, N, N'-carbonyldiimidazole, diphenylphosphoric acid azide, diethyl cyanophosphoric acid. It is also possible to use a peptide-forming reagent such as, for example, for condensation. In the condensation reaction, the compound (II),
The amidino group present in formula (II ′) or compound (IV) exists as a salt of an inorganic acid (eg, hydrogen chloride, sulfuric acid, nitric acid, hydrogen bromide), a tert-butoxycarbonyl group or a benzyloxycarbonyl group. It is preferably protected with a group. In any of the above cases, the condensation reaction is preferably the addition of an organic base (eg, triethylamine, N-methylpiperidine, 4-N, N-dimethylaminopyridine) or an inorganic base (sodium hydrogen carbonate, sodium carbonate, potassium carbonate). Can accelerate the reaction. The reaction temperature is generally -20 to + 50 ° C, preferably 0 ° C
~ Around room temperature, the solvent usually used is, for example, water,
Dioxane, tetrahydrofuran, acetonitrile, pyridine, N, N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone, chloroform, methylene chloride and the like can be mentioned, and they may be used alone or as a mixed solvent.

A carboxyl group-protecting group (benzyl group or tert-butyl group which is a carboxyl group-protecting group of X or Y in the general formula (I)) contained in the product of the final method and / or phenolic Protecting group for hydroxyl group or alcoholic hydroxyl group (in the general formula (I), R ¹⁰ , R ¹² , R ¹³
And a benzyl group which is a protective group for the hydroxyl group of X or tert-
The butyl group) can be removed by a method known per se. For example, a compound having a benzyl ester group or a benzyl ether group can be hydrogenated in the presence of a noble metal catalyst such as palladium or platinum to obtain a carboxylic acid derivative, or a tert-butyl ester group or a tert-butyl ether group. A compound having a can be treated with an acid such as trifluoroacetic acid or hydrogen chloride to obtain a carboxylic acid derivative. The salt of compound (I) is
Although it can be obtained by the reaction itself for producing the compound (I), it is also possible to produce a salt of the compound (I) as described above by adding an acid, an alkali or a base, if necessary.

The thus-obtained object compound (I) of the present invention is obtained from the reaction mixture by conventional means of separation and purification such as extraction, concentration, neutralization, filtration, recrystallization, column chromatography and thin layer chromatography. It can be isolated by Compound (I) can exist in at least two stereoisomers. All of these individual isomers and mixtures thereof are naturally included in the scope of the present invention, and if desired, these isomers can be individually produced. The above formulas (III) and (I
V), raw material compounds represented by (V), and the following formula (I
X), (X), (XII), (XIII), (XIV) and (XV)
A single optical isomer of compound (I) can be obtained by carrying out the reaction described above using a single isomer of each of the starting compounds represented by In the case of a mixture of isomers, this can be separated by a conventional separation method such as an optically active acid (eg, camphorsulfonic acid, tartaric acid, dibenzoyltartaric acid, etc.), an optically active base (eg, cinchonine, cinchonidine, quinine, quinidine, α-methylbenzyl). Amine, dehydroabiethylamine, etc.)
Each isomer can also be separated by a method of forming a salt with or by a separation means such as various chromatography and fractional recrystallization.

Raw material compounds (II) and (II ') of the present invention
Is a compound known per se, and the starting compounds represented by the formulas (III), (IV) and (V) can be produced according to a method known per se. For example, the method represented by the following reaction formula Can also be manufactured by. In the following description, the compound of formula (III) may be simply referred to as (III). Other compounds may be referred to in the same manner.

[0020]

[Chemical 9]

[Chemical 10]

In the above reaction formula, R is a protecting group for amino group of amino acid, and is a benzyloxycarbonyl group or tert.
-Butoxycarbonyl group and the like are shown. R ¹⁰ , R ¹² , R ¹³ and X may have a protective group for a phenolic hydroxyl group or an alcoholic hydroxyl group, and examples of such a protective group include a tert-butyl group and a substituted or unsubstituted benzyl group. . X or Y carboxyl group, benzyl group or tert-
It may have a protecting group such as a butyl group. The method for producing the compound (III) represented by the above reaction scheme will be described in more detail.
The reaction for obtaining III) is a normal alkylation reaction for amino groups, and compound (VI) and compound (VII) are combined with a base (eg, an inorganic base such as sodium carbonate, potassium carbonate, potassium hydrogen carbonate, or cesium fluoride or , An organic base such as triethylamine, pyridine, 4-N, N-dimethylaminopyridine, etc.), usually at a temperature of about 0 to + 100 ° C.
Examples of the reaction solvent include acetonitrile, N, N-dimethylformamide, tetrahydrofuran, toluene,
Examples include organic solvents such as methylene chloride. Then, the production of the compound (X) by the condensation reaction of the compound (VIII) and the N-protected derivative (IX) of the amino acid is a usual peptide bond forming reaction of the amino acid, and the compound (II) and the compound (II)
It can be produced under the same reaction conditions as the condensation reaction of I).

The cyclization reaction of the compound (X) thus obtained to the cyclic compound (XI) is an acid-catalyzed cyclization reaction. For example, as the catalyst, p-toluenesulfonic acid,
Organic sulfonic acids such as camphorsulfonic acid and methanesulfonic acid can be used as catalysts. The reaction is usually carried out by adding the compound (X) in a solvent such as toluene, benzene, ethyl acetate or 1,2-dichloroethane at a reaction temperature of 0 to +10.
Compound (XI) is obtained by reacting at 0 ° C, preferably +30 to + 80 ° C. Then, the reduction reaction from compound (XI) to compound (XII) is a reduction reaction of a double bond, for example, catalytic reduction using a metal such as platinum, palladium, Raney nickel or a mixture thereof with an arbitrary carrier, for example, It can be easily produced by reduction with a metal hydride compound such as sodium borohydride and sodium cyanoborohydride. The above reaction is usually carried out in the presence of an organic solvent (eg, methanol, ethanol, dioxane, ethyl acetate), and the reaction temperature is preferably −20 to + 100 ° C. although it varies depending on the reducing means. This reaction can sufficiently achieve the purpose at atmospheric pressure, but may be carried out under pressure depending on the circumstances. In this reaction, R represents a benzyloxycarbonyl group, and when the reduction reaction is carried out by a catalytic reduction reaction, the deprotection reaction of R also proceeds at the same time, and the compound (V) can be obtained all at once.

The deprotection reaction of (XII) ➝ (V) and (XIV) ➝ (III) is a usual reaction for removing the protecting group of the amino group of a peptide, and when R represents a benzyloxycarbonyl group, platinum is used. It can be removed by a catalytic reduction reaction using a metal such as palladium, rhodium, or the like as a catalyst. When R represents a tert-butoxycarbonyl group, it can be easily removed with an acid such as trifluoroacetic acid or hydrogen chloride in an organic solvent such as methanol, ethanol, ethyl acetate or dioxane. The condensation reaction between the compound (V) and the amino acid derivative (XIII) and the condensation reaction between the compound (II ′) and the compound (XV) are amide bond forming reactions,
It can be produced by a method similar to the condensation reaction of compound (II) and compound (III). The starting compound represented by the formula (XIII) is, for example, J. Am. Chem. Soc., 77 (19
It can be manufactured according to the method described in 55). More generally, compounds in which R ¹⁰ is an optionally substituted lower alkyloxy, lower alkenyloxy, lower alkynyloxy, lower alkanoyloxy, carbamoyloxy, or methanesulfonyloxy group (XII
I) is a benzyloxycarbonyl group or t
Tert or 3,4-dihydroxyphenylalanine (DOPA) protected by ert-butoxycarbonyl group
And dialkyl sulfuric acid such as dimethyl sulfate and diethyl sulfate,
Alternatively, it can be produced by reacting an alkenyl halide, an alkynyl halide, an acyl halide, an acid anhydride, an alkyl isocyanate, an alkyl sulfonyl halide and the like. The reaction is a base (eg, an inorganic base such as sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogencarbonate, cesium fluoride or an organic base such as triethylamine, pyridine, 4-N, N-dimethylaminopyridine). In the presence of
It can be performed at a temperature of about 0 to 100 ° C. Examples of the reaction solvent include water, acetonitrile, N, N-dimethylformamide, tetrahydrofuran, toluene, methylene chloride and the like.

The compound (XIII) in which R ¹⁰ is a lower alkynyl group, a lower alkenyl group or a lower alkyloxycarbonyl group can be produced according to a method known per se. For example, J. Chem. Soc., Chem. Commun., 904 (198)
7), J. Am. Chem. Soc., 109, 547 (198).
1), J. Org. Chem., 48, 3252 (1983). In the compound represented by the formula (XIII), R ¹⁰ is a nitro group or a halogen atom, and R ¹² ,
As the compound of the hydrogen atom of R ¹³ , known compounds which are commercially available can be used. Further, the compound generally represented by the formula (XIII) can also be produced by a method used as a method for synthesizing an α-amino acid. For example J. Am. Chem. So
c., 70, 1451 (1948), Proc. Chem. Soc.,
117 (1962), J. Am. Chem. Soc., 65, 221.
1 (1943). Further, each optical isomer of the compound represented by the formula (XIII) can be produced according to a method known in the literature. For example, U. S. P. , 3841966 (194
It can be manufactured according to the method described in 6).

Of the compounds represented by the formula (XIV), those in which R ¹⁰ is a formyl group are represented by the formula (XIV) in which R ¹⁰ is an allyl group.
It can be produced by oxidizing the compound (1) with ruthenium tetraoxide. Among the compounds represented by the formula (XIV), the compound in which R ¹⁰ is a hydroxymethyl group can be produced by reducing the compound (XIV) in which R ¹⁰ is a formyl group with sodium borohydride. Among the compounds represented by the formula (XIV), the compound in which Y is an esterified carboxyl group used as a prodrug can be produced according to a method known in the literature. That is, Chem. Pharm. Bul
l., 31, 2698 (1983), Chem. Pharm. Bul.
l., 32, 2241 (1984), J. Antibiotics, 4
0, 81 (1987), J. Antibiotics, 45, 135.
8 (1992) and the like. More generally, the compound (XIV) in which Y is a carboxyl group is described in Chem. Pharm. Bull., 31, 2698.
(1983), Chem. Pharm. Bull., 32, 2241.
(1984), J. Antibiotics, 40, 81 (198
7), J. Antibiotics. 45, 1358 (1992) and the like known organic halides and bases (eg, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, cesium fluoride, etc. Inorganic base or triethylamine, pyridine, 4-N, N-
It can be produced by reacting at a temperature of usually -20 to 100 ° C in the presence of an organic base such as dimethylaminopyridine. Examples of the reaction solvent include N, N-dimethylformamide, acetonitrile, methylene chloride, ethyl acetate, tetrahydrofuran and the like. Compound (I) above
In the process for producing the compound and its intermediate, the compound used in the reaction is an inorganic acid salt such as hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc., such as acetate, tartrate, etc., as long as the reaction is not hindered. , Citrate, fumarate, maleate, toluenesulfonate, methanesulfonate, and other organic acid salts such as sodium salt, potassium salt,
Metal salts such as calcium salt and aluminum salt, such as triethylamine salt, guanidine salt, ammonium salt,
It may be used in the form of a salt such as a salt with a base such as a hydrazine salt, a quinine salt or a cinchonine salt. When the free form of compound (I) is obtained by the above-mentioned production method, it is converted into its salt by a conventional method, and when it is obtained as a salt of compound (I), its salt is converted into a compound (I) by a conventional method. You can

Compounds of formula (I), including hydrates thereof
Is a low toxicity and safe compound that binds fibrinogen, fibronectin and von Willebrand factor to the platelet fibrinogen receptor (glycoprotein IIb / IIIa), and its equivalent on the surface of various types of cells. Inhibit the binding of those and other adhesive proteins such as vitronectin, collagen and laminin to the receptors. Therefore, the compounds of the present invention affect cell-cell and cell-matrix interactions, and in particular, the compounds interfere with platelet thrombin development, leading to peripheral arterial occlusion, acute myocardium in mammals, including humans. Infarction (AMI), deep vein thrombosis, pulmonary embolism, dissecting aneurysm, transient ischemic attack (TI)
A), stroke and other obstruction-related disorders, unstable angina,
Diffuse intravascular coagulation (DIC), sepsis, surgical or infectious shock, postoperative and postpartum trauma, cardiopulmonary bypass surgery, incompatible transfusion, placental ablation, thrombotic thrombocytopenic purpura (TTP), snake venom and It can be used for the treatment or prevention of acute or chronic renal diseases due to overaggregation such as immune diseases, inflammation, arteriosclerosis, hemolytic uremic syndrome, targeted peripheral necrosis, and pressure ulcers. Further, the compound (I) of the present invention enhances the action of a thrombolytic agent and prevents reocclusion, PTCA
It can be used for prevention of subsequent reocclusion, prevention of thrombocytopenia due to dialysis, prevention of blood clots caused by artificial blood vessels and organs, and can also be used as an antitumor agent by inhibiting cancer metastasis.

Medical compositions containing compounds of formula (I), including their hydrates and their salts, are for example tablets, lacquered tablets, sugar-coated tablets, hard and soft gelatin capsules, solutions, emulsions. It can be administered orally in the form of solutions or suspensions, rectally in the form of suppositories or as a spray. However, the administration also
It can also be carried out parenterally, for example in the form of injectable solutions. The above-mentioned preparations in each form can be produced according to a conventional method, if necessary, using an excipient and the like. tablet,
The active compounds can be mixed with pharmaceutically inert, inorganic or organic excipients for the production of lacquered tablets, sugar-coated tablets and hard gelatine capsules. tablet,
Representative of such excipients which can be used for sugar coated tablets and hard gelatin capsules are lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof. Examples of suitable excipients for soft gelatin capsules are vegetable oils, waxes, fats,
Semi-solid and liquid polyols. However,
If the properties of the active compound are suitable, then no excipient is necessary when using soft gelatin capsules.

Examples of suitable excipients for the production of solutions and syrups are water, polyols, sucrose, invert sugar and glucose. Suitable examples for injectable solutions are water, alcohols, polyols, glycerol and vegetable oils. Suitable examples for suppositories are natural or hardened oils, waxes, fats and semi-liquid or liquid polyols. The pharmaceutical composition further comprises preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavors, osmotic pressure changing salts, buffers, coatings or antioxidants. can do. The dose of the active compounds which suppresses or prevents the abovementioned diseases can be varied within wide limits. And of course the adjustments have to be made to suit the individual circumstances in each particular case. Generally, about 0.01-20 mg / kg per day,
Preferably a dose of about 0.1-4 mg / kg is suitable for oral administration to adults. In the case of parenteral administration, the daily dose for an adult is usually about 0.005-1.0 mg / kg, preferably about 0.01-0.3 mg / kg.

[0029]

INDUSTRIAL APPLICABILITY The present invention provides compounds and pharmaceuticals that are effective in preventing or treating various diseases by regulating or inhibiting cell adhesion. In particular, the compound of the present invention exerts an inhibitory action on platelet aggregation by ADP for a long period of time by oral administration, so that it can be a stronger and more durable antithrombotic agent than known agents having the same effect.

[0030]

EXAMPLES The present invention will be described in more detail with reference to test examples, reference examples and examples, but the present invention is not limited thereto. Test Example 1 In vitro binding experiment

Method: Washed platelets were prepared from citrated blood of guinea pigs and suspended in Hepes-Tyrode solution (pH 7.5) to adjust the platelet count to 500,000 / μl. This platelet suspension 70
10 μl of ADP solution (final concentration: 5 × 10 ⁻⁵ M) was added to μl and left at room temperature for 15 minutes. Test drug solution and
¹²⁵ I-fibrinogen solution (final concentration: 0.1 μM) 80
μl was added and left at room temperature for 2 hours. Reaction solution 100 μl
Was overlaid with 20% sucrose buffer and centrifuged at 3,000 rpm for 20 minutes to perform ¹²⁵ I-fib
The rinogen was isolated. Platelet binding ¹²⁵ I-fibrinogen
Was measured, and the binding inhibition rate was determined from the value of the test drug treated group relative to the specific binding of the control group (buffer solution treated). Effect of the compound of Example 1 on the specific binding of ¹²⁵ I-fibrinogen to guinea pig-activated platelets IC ₅₀ value: 2.7 × 10 ⁻⁹ M (n = 3; where n is the number of experimental animals) (In the following test examples, n is synonymous)

Test Example 2 In vitro measurement of platelet aggregation

[Method] In the case of human, the guinea pig was anesthetized with pentobarbital (20 mg / kg, ip) from the brachial vein of a healthy male who had not taken the drug for more than 2 weeks, and sodium citrate was used as an anticoagulant from the abdominal aorta (final). Concentration: 0.38
%, Human; 0.315%, guinea pig) Blood was collected. Citrated blood at 950 rpm for 10 minutes and 2,000 rpm
10 minutes each at room temperature by centrifugation to obtain platelet rich plasma (PRP) and platelet poor plasma (PPP).
RP was diluted with PPP to obtain a platelet count of 500,000 / μl
Was prepared. Platelet aggregation is the method of Born (Nature 19
4: 927, 1962), and was measured spectroscopically using an aggregometer (Hematracer 6, Toa Medical Electronics). After incubating PRP (250 μl) at 37 ° C. for 2 minutes, 25 μl of the test drug solution was added, and 2 minutes after that, 25 μl of an aggregation inducer was added. The effect of the test drug was represented by the inhibition rate of the maximum aggregation rate of the test drug treatment group with respect to the maximum aggregation rate of the control group. ADP used the lowest concentration that yielded maximum aggregation (0.6-1 μM). Inhibitory Action of Example Compounds on ADP-induced Platelet Aggregation ───────────────────────────────────────────────────────────────────────────── ₅₀ value (M) Example number Human (n = 3) Guinea pig (n = 3) ─────────────────────────────── ^{1 2.0 × 10 -8 3.7 × 10} -8 3 1.0 × 10 -8 1.5 × 10 -8 4 - 1.2 × 10 -7 5 4.0 × 10 -8 3.4 × 10 -8 14 1.6 × 10 -8 3.3 × 10 - ^{8 18 2.0 × 10 -8 2.6 ×} 10 -8 19 2.1 × 10 -8 2.2 × 10 -8 21 2.6 × 10 -8 3.8 × 10 -8 22 1.8 × 10 -8 4.7 × 10 -8 23 1.7 × 10 - ⁸ 3.8 × 10 ^-8 24 2.4 × 10 ^-8 4.6 × 10 ^-8 31 1.2 × 10 ^-8 1.8 × 10 ^-8 ─────────────────────── ──────── (-Not measured)

Test Example 3 Ex vivo platelet aggregation experiment

Method: The test drug solution was orally administered (2 ml / kg) to guinea pigs that had been fasted overnight, and blood was collected under pentobarbital anesthesia (20 mg / kg, ip) 1, 4 and 24 hours later.
PRP was prepared by the method described above, the platelet aggregation by ADP was measured, and the inhibition rate was calculated from the aggregation rate of the test drug administration group relative to the aggregation rate of the control group. The test drug was dissolved in distilled water and orally administered. ADP aggregation inhibitory effect of the compound of Example 1 in guinea pigs 3 mg / kg oral administration 1 hour later Inhibition rate: 100% (n
= 4) 3 mg / kg Oral administration 4 hours later Suppression rate: 100% (n
= 4) 3 mg / kg, 24 hours after oral administration Suppression rate: 100% (n
= 3) ADP aggregation inhibitory action of the compound of Example 3 in guinea pig 1 mg / kg Oral administration 1 hour later Inhibition rate: 95% (n
= 6) 1 mg / kg Oral administration 4 hours later Suppression rate: 100% (n
= 6)

Reference Example 1 N- (2,2-dimethoxyethyl) glycine tert-butyl ester 150 g of 2,2-dimethoxyethylamine, 200 g of anhydrous potassium carbonate and 1.4 liter of N, N-dimethylformamide were added to chloroacetic acid tert. 100 ml of butyl ester are added with stirring at 20-25 ° C. After stirring for 2 hours, 100 ml of chloroacetic acid tert-butyl ester was added, and the mixture was stirred at 20-25 ° C for 7 hours. Ice water 1.4 liters is added to the reaction solution, and the mixture is extracted with ethyl acetate. The organic layer is washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent is evaporated under reduced pressure. The residue is subjected to vacuum distillation to obtain a fraction having a boiling point of 80-85 ° C (0.4 mmHg) as a target product. Yield 163g. NMR spectrum (CDCl ₃ ) δ: 1.49 (s, 9H), 2.75
(d, J = 5.4Hz, 2H), 3.33 (s, 2H), 3.39 (s, 6H), 4.47 (t, J =
(5.4Hz, 1H)

Reference Example 2 3-Methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester oxalate N- (2,2-dimethoxyethyl) glycine tert-butyl ester 44 g and N-benzyloxycarbonyl- L-aspartic acid β-methyl ester 49.5
44 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide was added to a solution prepared by dissolving g in 400 ml of methylene chloride while stirring under ice cooling. Stir under ice cooling for 10 minutes, then at room temperature for 50 minutes. Water 30 in the reaction solution
Add 0 ml and 100 ml of 5% KHSO ₄ aqueous solution and shake. The organic layer is separated and the aqueous layer is extracted with 200 ml methylene chloride. The organic layers are combined, washed with water, dried over anhydrous sodium sulfate, and the solvent is distilled off under reduced pressure. The resulting residue (8
8 g) is dissolved in 1 liter of toluene, 4.0 g of p-toluenesulfonic acid is added, and the mixture is stirred at 70-75 ° C. for 4 hours. After cooling, the reaction solution is washed with an aqueous sodium hydrogen carbonate solution, dried over anhydrous sodium sulfate, and the solvent is distilled off under reduced pressure. The residue was subjected to column chromatography on silica gel (600 ml), eluent (ethyl acetate-hexane = 1: 1).
Fractions eluted at 1 liter are collected and concentrated under reduced pressure. To a solution of the obtained residue (68 g) in ethyl acetate (1.5 liter) was added 10% palladium-carbon (16 g), and the mixture was vigorously stirred in a hydrogen stream for 3 hours. A solution prepared by dissolving 20.2 g of oxalic acid dihydrate in 200 ml of methanol is added to the filtrate obtained by removing the catalyst, and the mixture is left to stand. The precipitated crystals are collected by filtration,
After washing with ethyl acetate and drying, 46.8 g of the desired compound is obtained. Mp 146-147 ° C. Specific rotation [α] ^{_{D 23 -26.0 ° (c = 0.603}} , methanol) Elemental _{analysis: C 13 H 22 N 2 O} 5 · C 2 H 2 O 4 Calculated: C, 47.87; H, 6.43; N, 7.44 Found: C, 47.79; H, 6.20; N, 7.42

Reference Example 3 (S) -4-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester acetate (S) -3-methoxycarbonylmethyl-2-oxopiperazine -1-acetic acid tert-butyl ester 1.65 g,
N-benzyloxycarbonyl-L-tyrosine 1.9 g
Was dissolved in 15 ml of dichloromethane, 1.44 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added with stirring at room temperature, and the mixture was stirred for 1 hour. The reaction solution is concentrated under reduced pressure, the resulting oily substance is dissolved in ethyl acetate, the organic layer is washed with 5% potassium hydrogen sulfate, dried over anhydrous magnesium sulfate, and the solvent is distilled off under reduced pressure. The obtained residue was subjected to silica gel column chromatography (eluent: ethyl acetate: hexane = 7: 3),
2.0 g of a colorless oily product of (S) -4- (N-benzyloxycarbonyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester is obtained. IR ν _max cm ^-1 : 3332, 2952, 1740, 1646, 1613, 151
4, 1444, 1367, 1227, 1153. NMR (CDCl ₃ ) δ: 1.46 (9H, s), 2.30-2.53 (1H, m),
2.72-3.13 (5H, m), 3.50-3.72 (2H, m), 3.63 (3H, s), 3.8
5 (1H, d, J = 12Hz), 4.07 (1H, d, J = 12Hz), 4.70-4.90 (1H,
m), 4.90-5.05 (1H, m), 5.11 (2H, s), 5.50-5.67 (2H, m),
6.76 (2H, d, J = 8.2Hz), 7.03 (2H, d, J = 8.2Hz), 7.36 (5H,
s). Obtained (S) -4- (N-benzyloxycarbonyl-L-tyrosyl) -3-methoxycarbonylmethyl-2
-Oxopiperazine-1-acetic acid tert-butyl ester 1.3 g, 10% palladium-carbon 300 mg, acetic acid 18
A mixture of 0 mg and 20 ml of methanol was added in a hydrogen stream at room temperature.
Stir for 0 minutes. The catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to give (S) -4-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester acetate colorless oil 1 To obtain 0.0 g. This product can be used in the next reaction without purification.

Example 1 (S) -4- (4-amidinobenzoyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid hydrochloride (S) -obtained in Reference Example 3 4-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-
1.0 g of acetic acid tert-butyl ester and 380 mg of sodium hydrogen carbonate are dissolved in a mixture of 20 ml of water and 10 ml of dioxane and 505 mg of 4-amidinobenzoyl chloride hydrochloride are added over 10 minutes with vigorous stirring. After stirring for 1 hour, the reaction solution is concentrated under reduced pressure. To the resulting residue was added 10 ml of trifluoroacetic acid and 1
Stir for hours. The reaction solution was concentrated to dryness under reduced pressure, the residue was dissolved in 5 ml of 1N hydrochloric acid, and applied to a Wakogel LP-C18 column (eluent: 7.5% acetonitrile aqueous solution),
Fractions containing the target compound are collected and freeze-dried to obtain 550 mg of the target compound as a colorless powder. Specific rotation: [α] _D ²³ + 91 ° (c = 0.2, H ₂ O) Elemental analysis value: C ₂₆ H ₂₉ N ₅ O ₈ · HCl (576.0) Calculated value: C, 54.22; H , 5.25; N, 12.16 Found: C, 53.94; H, 5.31; N, 11.93.

Reference Example 4 (S) -4-O- ^t- Butyl-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid
tert- butyl ester acetate salt (3) - methoxycarbonylmethyl-2-oxo-piperazin-1-acetic acid tert- butyl ester 1.87 g, and N- benzyloxycarbonyl -O- ^t-butyl -L
-Treating 2.43 g of tyrosine as in Reference Example 3,
(S) -4- (N-benzyloxycarbonyl-O- ^t
3.2 g of a colorless oil of butyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester are obtained. IR ν _max cm ^-1 : 3300, 2980, 1740, 1655, 1505, 144
0, 1365, 1237, 1160. NMR (CDCl ₃ ) δ: 1.31 (9H, s), 1.46 (9H, s), 2.35
-2.51 (1H, m), 2.70-3.18 (5H, m), 3.57-3.70 (2H, m), 3.6
3 (3H, s), 3.81 (1H, d, J = 17Hz), 4.02 (1H, d, J = 17Hz), 4.7
5-4.88 (1H, m), 5.00 (1H, t, J = 5.2Hz), 5.10 (2H, s), 5.65
(1H, d, J = 8.8Hz), 6.91 (2H, d, J = 8.4Hz), 7.10 (2H, d, J = 8.4Hz
Hz), 7.35 (5H, s ). The resulting (S) -4- (N- benzyloxycarbonyl -O- ^t-butyl -L- tyrosyl) -3-methoxycarbonylmethyl-2-oxo-piperazine-1-acetic acid tert-butyl ester 3.2 g, 10% palladium carbon 1.0
g, 300 mg of acetic acid and 50 ml of methanol are stirred in a stream of hydrogen for 30 minutes. Remove the catalyst by filtration,
The filtrate is concentrated under reduced pressure to obtain 2.80 g of the target colorless oil. This product can be used as it is in the next reaction.

Example 2 (S) -4- (4-amidinobenzoyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid (S) -4-obtained in Reference Example 4 2.80 g of O- ^t- butyl-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid acetate, 840 mg of sodium hydrogencarbonate were dissolved in a mixed solution of 40 ml of water and 20 ml of dioxane, and the mixture was stirred 4 vigorously. 1.1 g amidinobenzoyl chloride hydrochloride are added in 20 minutes. The reaction solution is concentrated under reduced pressure, the resulting crude product is dissolved in 30 ml of trifluoroacetic acid, left for 1 hour and then concentrated under reduced pressure. The residue is purified on a MCI GEL CHP20P column (10% aqueous acetonitrile). Concentrate the target fraction,
Recrystallization of the obtained residue from water-methanol gives 1.9 g of colorless crystals of the desired compound. Mp 229-235 ° C. Specific rotation: [α] ^{_{D 20 + 82 ° (c =}} 1.0, H 2 O) Elemental _{analysis: C 26 H 29 N 5 O} 8 · 3 / 2H 2 O Calculated: C, 55.12; H, 5.69; N, 12.36 Found: C, 55.38; H, 5.66; N, 12.40.

Reference Example 5 (S) -4-O-methyl-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid
tert-Butyl ester acetic acid salt (S) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester obtained in Reference Example 2 and N-benzyloxycarbonyl-O-methyl-L
-Tyrosine was condensed in the same manner as in Reference Example 3 and then catalytically reduced (S) -4-O-methyl-L-tyrosyl-3.
-Methoxycarbonylmethyl-2-oxopiperazine-
A colorless oil of 1-acetic acid tert-butyl ester acetate is obtained. Yield 85%. By the same method, the following Reference Example compound 6-9 was obtained as a colorless oil.

[Table 1]

Reference Example 10 (S) -4-L-3,4-dimethoxyphenylalanyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid trifluoroacetate salt Obtained in Reference Example 2 (S). -3-Methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester is condensed with N-tert-butoxycarbonyl-L-3,4-dimethoxyphenylalanine and then reacted with trifluoroacetic acid in methylene chloride. Then, the solvent is distilled off under reduced pressure to obtain a colorless oily substance of the target compound. This product can be used in the next reaction without purification. Reference Example 11 (S) -4-L-3,4-diethoxyphenylalanyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid trifluoroacetate salt In the same manner as in Reference Example 10 (S). -3-Methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-
Butyl ester and N-tert-butoxycarbonyl-L-
It can be produced from 3,4-diethoxyphenylalanine. Reference Example 12 (S) -3-tert-butoxymethyl-2-oxopiperazine-1-acetic acid tert-butyl ester oxalate N- (2,2-dimethoxyethyl) glycine tert-butyl ester and N-benzyloxycarbonyl -O-te
It was synthesized from rt-butyl-L-serine by the same method as in Reference Example 2. Mp 178-180 ° C. Specific rotation [α] ^{_{D 20 -30.4 ° (c = 1.0}} , DMS
O) Elemental analysis _{C 15 H 28 N 2 O 4} · C 2 H 2 O 4 · 0.5H 2 O
Calculated as: C, 51.12; H, 7.82; N, 7.01 Found: C, 51.40; H, 7.71; N, 6.94

Reference Example 13 (S) -3- (4-tert-Butoxybenzyl) -2-oxopiperazine-1-acetic acid tert-butyl ester N- (2,2-dimethoxyethyl) glycine and N-benzyloxycarbonyl It was synthesized from —O-tert-butyl-L-tyrosine by the same method as in Reference Example 2. Mp 124-126 ° C. Specific rotation [α] ^{_{D 20 -81.7 ° (c = 0.97}} , CHCl
₃ ) Elemental analysis value C ₂₁ H ₃₂ N ₂ O ₄ Calculated value: C, 66.99; H, 8.57; N, 7.44 Measured value: C, 66.92; H, 8.54; N, 7.38 In Reference Example 12 and Reference Example 13 The obtained compound and N-benzyloxycarbonyl-O-tert-butyl-L-tyrosine are condensed in the same manner as in Reference Example 3, and then catalytically reduced to obtain the compounds of Reference Examples 14 and 15. Reference Example 14 (S) -4-O-tert-butyl-L-tyrosyl-3-te
rt-Butoxymethyl-2-oxopiperazine-1-acetic acid
tert-Butyl ester acetate Reference Example 15 (S) -4-O-tert-butyl-L-tyrosyl-3-
(4-tert-Butoxybenzyl) -2-oxopiperazine-1-acetic acid tert-butyl ester acetate

Example 3 (S) -4- (4-amidinobenzoyl-O-methyl-)
L-tyrosyl) -3-methoxycarbonylmethyl-2-
Oxopiperazine-1-acetic acid (S) -4-O-methyl-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester obtained in Reference Example 5 and 4-amidinobenzoyl chloride And were condensed in the same manner as in Example 2, followed by deprotection with trifluoroacetic acid, and then MCI
Purification with a GEL CHP20P column (Mitsubishi Kasei) (eluent H ₂ O → 5% CH ₃ CN → 10% CH ₃ CN) and recrystallization from water-ethanol give colorless crystals of the target compound. 66% mp 208-212 ° C. Specific rotation yield [α] ^{_{D 20 -76.7 ° (c = 1.035}} , DM
SO) Elemental analysis value Calculated as C ₂₇ H ₃₁ N ₅ O ₈ · 3H ₂ O: C, 53.37; H, 6.14; N, 11.53 Actual value: C, 53.15; H, 6.14; N, 11.36 By the same method Reference examples 5, 6, 7, 8, 9, 10, 11,
The compounds obtained in Examples 14 and 15 and 4-amidinobenzoyl chloride were treated in the same manner as in Example 3 to give Example 4-
11 compounds were obtained.

[0043]

[Table 2]

[0044]

[Table 3]

By the same method as in Reference Example 5, Reference Example compound 15-20 was obtained as a colorless oil.

[0046]

[Table 4] The following Reference Example compound 2 was prepared in the same manner as in Reference Example 10.
1-26 was obtained as a colorless oil.

[0047]

[Table 5] The compounds obtained in Reference Examples 15, 16, 17, 18, 19, and 20 and 4-amidinobenzoyl chloride were subjected to the same operations as in Example 3 to obtain the following compounds of Examples 12-17.

[0048]

[Table 6] The compounds obtained in Reference Examples 21, 22, 23, 24, 25 and 26 and 4-amidinobenzoyl chloride were subjected to the same operations as in Example 2 to obtain the following compounds of Examples 18-23.

[0049]

[Table 7]

Example 24 (S) -4- {4-amidinobenzoyl- (3,4-dihydroxy) -DL-phenylalanyl} -3-methoxycarbonyl-2-oxopiperazine-1-acetic acid In Reference Example 2 Obtained (S) -tert-butyl 3-methoxycarbonylmethyl-2-oxopiperazine-acetic acid oxalate, N-benzyloxycarbonyl-3,4-di (methoxymethyloxy) -DL-phenylalanine,
Using 4-amidinobenzoyl chloride and the same operations as in Reference Example 5 and Example 3, the title compound was obtained as a colorless amorphous powder. Specific rotation [α] _D ²⁰ 64.2 ° (C = 0.5, H ₂ O) Elemental analysis value: C ₂₆ H ₂₉ N ₅ O ₉ · 2H ₂ O Calculated value: C, 52.79; H, 5.62; N, 11.84. Found: C, 52.83; H, 5.75; N, 11.66. Example 25 (S) -4- {4-amidinobenzoyl- (4-chloro) -DL-phenylalanyl} -3-methoxy. Carbonyl-2-oxopiperazine-1-acetic acid (S) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid tert-butyl ester obtained in Reference Example 2, N-benzyloxycarbonyl-4-chloro-D
The title compound was obtained as a colorless amorphous powder by the same procedures as in Reference Example 10 and Example 2 using L-phenylalanine and 4-amidinobenzoyl chloride. Specific rotation [α] _D ²⁰ 56.8 ° (c = 0.45, H
₂ O) Elemental _{analysis: C 26 H 28 ClN 5 O} 7 · H 2 O Calculated:. C, 54.22; H, 5.25; N, 12.16 Found: C, 53.98; H, 5.31 ; N, 12.33.

Reference Example 27 Ethyl N- ^t -butoxycarbonyl-L-tyrosinate Ethyl L-tyrosinate hydrochloride (25.0 g) and sodium hydrogencarbonate (9.41 g) were dissolved in water (125.0 ml) and 1,4-dioxane (125.0 ml). With stirring at room temperature, 25.7 ml of di- ^t- butyl dicarbonate was added. After stirring for 1 hour, liquid separation extraction was performed with methylene chloride, and the organic layer was concentrated under reduced pressure. The residue was dissolved in a small amount of ethyl acetate and crystallized from petroleum ether to give the title compound (29.0 g, 91.8%) as white crystals. Melting point: 83-86 ° C. Specific optical rotation [α] _D ²⁰ : + 41.7 ° (C = 1.0, CHC
l ₃ ) Elemental analysis value: C ₁₆ H ₂₂ NO ₅ · 0.1H ₂ O Calculated value: C, 61.96; H, 7.21; N, 4.52 Analysis value: C, 61.81; H, 7.41; N, 4.51 Reference Example 28 Ethyl N- ^t -butoxycarbonyl-4-trifluoromethanesulfonyloxy-L-phenylalanate Ethyl N- ^t -butoxycarbonyl-L-tyrosinate 6.0 g, 2,6-lutidine 3.40 ml, and 4-dimethylaminopyridine 0. 48 g was dissolved in 97.3 ml of methylene chloride, 4.91 ml of trifluoromethanesulfonic anhydride was slowly added dropwise with stirring at -30 ° C, and then the mixture was stirred at room temperature for 1 hour. Water was added to the reaction system, liquid separation extraction was performed with methylene chloride, and the organic layer was concentrated under reduced pressure. The residue was roughly purified by silica gel chromatography (hexane / ethyl acetate = 4/1) and crystallized from (ethyl acetate / pentane) to give the title compound (4.9 g, 57.2%) as pale red crystals. . Mp: 49-50 ° C. Specific rotation [α] ^{_{D 20: + 32.3 ℃ (C}} = 1.0, CHC
l ₃ ) Elemental analysis value: C ₁₇ H ₂₁ F ₃ NO ₇ S Calculated value: C, 46.36; H, 4.81; N, 3.18 Analysis value: C, 46.52; H, 5.07; N, 3.13

Reference Example 29 Ethyl N- ^t Butoxycarbonyl-4-vinyl-L-
Phenylalanate Ethyl N- ^t -butoxycarbonyl-4-trifluorone methanesulfonyloxy-L-phenylalanate 2.89 g, lithium chloride 834 mg, bis (triphenylphosphine) palladium (II) chloride 92.1 mg, 2,
145 mg of 6-di- ^t- butyl-4-methylphenol and 1.99 ml of vinyltributyltin were dissolved in 49.1 ml of N, N-dimethylformamide, and the mixture was dissolved in nitrogen atmosphere at 90 ° C. for 2 hours.
Stir for hours. 3.31 ml of pyridine and 6.61 ml of pyridinium fluoride (1.4 M tetrahydrofuran solution) were added to the reaction system, and the mixture was further stirred at room temperature for 18 hours. Water was added to the reaction system to remove insoluble matter by filtration, the mixture was extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure. The residue was purified by silica gel chromatography (hexane / ethyl acetate = 4/1) to give the title compound (1.65 g, 78.5%) as a pale-yellow oil. IR ν max cm ^-1 : 2978, 2932, 1738, 1713, 1510, 1444, 1
390, 1366, 1346, 1249, 1168, 1095, 1056, 989, 906,
850, 828, 777 NMR (CDCl ₃ ) δ: 1.24 (t, J = 7.2Hz, 3H), 1.42 (s, 9H), 2.90
-3.21 (m, 2H), 4.17 (q, J = 7.2Hz, 2H), 4.47-4.63 (m, 1H),
4.90-5.08 (m, 1H), 5.23 (d, J = 10.8Hz, 1H), 5.72 (d, J = 17.
6Hz, 1H), 6.69 (dd, J = 17.6Hz, J = 11.0Hz, 1H), 7.10 (d, J =
8.0Hz, 2H), 7.34 (d, J = 8.0Hz, 2H)

Reference Example 30 N- ^t -butoxycarbonyl-4-vinyl-L-phenylalanine ethyl N- ^t -butoxycarbonyl-4-vinyl-L-
Phenylalanate 1.65g and lithium hydroxide-1
0.24 g of the hydrate was dissolved in 5.17 ml of a mixed solvent of methanol: water = 10: 1 and stirred at room temperature for 20 minutes. The reaction system was adjusted to pH 2-3 with 1N HCl water, and separated and extracted with ethyl acetate. The organic layer was concentrated under reduced pressure to give the title compound 1.4.
0 g (92.7%) was obtained as a colorless oil. IR ν max cm ^-1 : 3428, 2978, 1714, 1511, 1442, 1393, 1
366, 1249, 1164, 1055NMR (CDCl ₃ ) δ: 1.42 (s, 9H), 2.9
9-3.28 (m, 2H), 4.53-4.70 (m, 1H), 4.83-5.04 (m, 1H), 5.
23 (d, J = 10.8Hz, 1H), 5.73 (d, J = 17.6Hz, 1H), 6.69 (dd, J =
11.0Hz, J = 17.6Hz, 1H), 6.40-7.00 (br, 1H), 7.14 (d, J = 8.
2Hz, 2H), 7.35 (d, J = 8.2Hz, 2H) Reference Example 31 (S) -4- (N- ^t -butoxycarbonyl-4-vinyl-L)
-Phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester (S) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester oxalate 2.17
g, and N- ^t -butoxycarbonyl-4-vinyl-L-
Phenylalanine (1.40 g) was dissolved in methylene chloride (21.7 ml), and while cooling with water, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.38 g) was added to 5 ml.
Add slowly over minutes. After stirring at room temperature for 1 hour, methylene chloride was distilled off under reduced pressure. The residue was dissolved in ethyl acetate, washed with 5% aqueous potassium hydrogen sulfate solution and saturated aqueous sodium hydrogen carbonate solution, the organic layer was dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue (ethyl acetate /
Crystallization with petroleum ether) gave the title compound 2.04 g (7
5.8%) was obtained as white crystals. Mp: 148-152 ° C. Specific rotation [α] ^{_{D 20: + 94.3 ° (C}} = 0.5%, CH
Cl ₃ ) Elemental analysis value: C ₂₉ H ₄₁ N ₃ O ₈ Calculated value: C, 62.24; H, 7.38; N, 7.51 Analysis value: C, 62.01; H, 7.51; N, 7.47

Example 26 (S) -4- (4-amidinobenzoyl-L-4-vinylphenylalanyl) -3-methoxycarbonylmethyl-
2-oxopiperazine-1-acetic acid (S) -4- (N- ^t -butoxycarbonyl-4-vinyl-L)
-Phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester 0.
60 g of methylene chloride 3.0 ml, trifluoroacetic acid 3.0
It was dissolved in ml and stirred at room temperature for 1 hour. The reaction system was concentrated under reduced pressure and azeotropically distilled several times with toluene. This product is H ₂ O 11.
It was dissolved in a mixed solvent of 1 ml and 1,4-dioxane (5.6 ml), and 0.45 g of sodium hydrogen carbonate was added at room temperature.
0.35 g of 4-amidinobenzoyl chloride at room temperature
Added over minutes. After stirring for 1 hour, with 1N HCl water
After adjusting to pH 2-3, the reaction solution was concentrated under reduced pressure. The residue is C
HP-20 column (H ₂ O → 5% CH ₃ CN / H ₂ O → 10
% CH ₃ CN / H ₂ O → 15% CH ₃ CN / H ₂ O → 20%
CH ₃ CN / H ₂ was purified by O), DAIKI compound and lyophilized converted to the hydrochloride salt by 1N HCl aqueous 0.39 g (58.
4%) was obtained as a colorless amorphous powder. Specific rotation [α] _D ²⁰ : + 84.9 ° (C = 1.0%, H ₂
O) Elemental _{analysis: C 28 H 32 N 5 O} 7 Cl · 2.0H 2 O Calculated: C, 54.06; H, 5.83 ; N, 11.26 analysis: C, 53.82; H, 5.52 ; N, 11.11

Example 27 (S) -4- (4-amidinobenzoyl-L-4-ethynylphenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ethyl N- ^t -butoxycarbonyl-4 -Trifluoromethanesulfonyloxy-L-phenylalanate 2.
97 g, lithium chloride 857 mg, bis (triphenylphosphine) palladium (II) chloride 94.6 mg, 2.6
Di- ^t- butyl-4-methylphenol (149 mg) and ethynyltributyltin (2.03 ml) were dissolved in dimethylformamide (50.5 ml), and the mixture was stirred at 90 ° C for 2 hours under a nitrogen atmosphere. Pyridine 4.8 ml and pyridinium fluoride 9.6 ml (1.4 M tetrahydrofuran solution) were added to the reaction system, and the mixture was further stirred at room temperature for 18 hours. Water was added to the reaction system to remove insoluble matter by filtration, the mixture was extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure. The residue was purified by silica gel chromatography (hexane / ethyl acetate = 4/1). This product and 34.3 mg of lithium hydroxide monohydrate were dissolved in 0.74 ml of a mixed solvent of methanol: water = 10: 1, and the mixture was stirred at room temperature for 45 minutes. PH of the reaction system with 1N HCl water
It was made 2-3, and the layers were separated and extracted with ethyl acetate. The organic layer was concentrated under reduced pressure to obtain N- ^t -butoxycarbonyl-4-ethynyl-L-phenylalanine as a colorless oil. This product and (S) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester oxalate 0.4
2 g was dissolved in 4.2 ml of methylene chloride, and 0.24 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was slowly added over 5 minutes while cooling with water. After stirring at room temperature for 1 hour, methylene chloride was distilled off under reduced pressure. The residue was dissolved in ethyl acetate, washed with 5% aqueous potassium hydrogen sulfate solution and saturated aqueous sodium hydrogen carbonate solution, the organic layer was dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (hexane / ethyl acetate = 2/3), and (S) 4- (N-
370 mg of ^t -butoxycarbonyl-4-ethynyl-L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester was obtained as a colorless oil. This product was dissolved in 1.9 ml of methylene chloride and 1.9 ml of trifluoroacetic acid and stirred at room temperature for 1 hour.
The reaction system was concentrated under reduced pressure and azeotropically distilled several times with toluene. The residue was dissolved in a mixed solvent of 6.8 ml of H ₂ O and 3.4 ml of 1,4-dioxane, 0.28 g of sodium hydrogencarbonate was added at room temperature, and 0.22 g of 4-amidinobenzoyl chloride was added at room temperature over 5 minutes. added. After stirring for 1 hour, the pH was adjusted to 2-3 with 1N HCl water, and the reaction solution was concentrated under reduced pressure.
P-20 column (H ₂ O → 5% CH ₃ CN / H ₂ O → 10%
CH ₃ CN / H ₂ O → 15% CH ₃ CN / H ₂ O → 20% C
H ₃ CN / H ₂ O) and lyophilized as a hydrochloride with 1N HCl water, 0.13 g (3.0%) of the title compound.
Was obtained as a colorless amorphous powder. Specific rotation [α] _D ²⁰ : + 57.2 ° (C = 1.0, H
₂ O) Elemental analysis value: C ₂₈ H ₃₀ N ₅ O ₇ Cl.3.0H ₂ O Calculation value: C, 52.71; H, 5.69; N, 10.98 Analysis value: C, 52.61; H, 5.51; N, 11.06

Reference Example 32 ^t- Butyl N-benzyloxycarbonyl-L-tyrosinate 5.0 g of ^t- butyl L-tyrosinate and 2.65 g of sodium hydrogen carbonate were added to 25.0 ml of H ₂ O and 1,4-dioxane 2
It was dissolved in 5.0 ml, and 4.51 ml of benzyloxycarbonyl chloride was added with stirring at room temperature. After stirring for 15 minutes, liquid separation extraction was performed with ethyl acetate. The organic layer was concentrated under reduced pressure,
The residue was purified by silica gel chromatography (hexane / ethyl acetate = 2/1) to obtain 7.79 g of the title compound as a pale yellow oily substance. IRνmax cm ^-1 : 3346, 2978, 1700, 1613, 1594, 1514, 1
452, 1366, 1227, 1153, 1103, 1057, 842, 751, 696 NMR (CDCl ₃ ) δ: 1.41 (s, 9H), 2.83-3.10 (m, 2H), 4.48 (d
d, J = 6.2Hz, J = 14.4Hz, 1H), 5.08 (s, 2H), 5.31 (d, J = 8.2Hz,
1H), 6.22 (s, 1H), 6.69 (d, J = 8.4Hz, 2H), 6.97 (d, J = 8.4H
z, 2H), 7.32 (s, 5H) Reference Example 33 ^t- butyl N-benzyloxycarbonyl-4-trifluorone methanesulfonyloxy-L-phenylalanate ^t- butyl N-benzyloxycarbonyl-L-tyrosinate 15.6 g, 7.36 ml of 2,6-lutidine, and 4
-Dimethylaminopyridine 1.03 g with methylene chloride 2
The solution was dissolved in 10 ml and 10.6 ml of trifluoromethanesulfonic anhydride was slowly added dropwise at -30 ° C with stirring, and then the mixture was stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with methylene chloride, and the organic layer was concentrated under reduced pressure. The residue was chromatographed on silica gel (hexane / ethyl acetate = 4
/ 1) to obtain 11.39 g (53.7%) of the title compound. IR ν max cm ^-1 : 3380, 1741, 1697, 1530, 1501, 1419, 1
369, 1348, 1249, 1226,1143, 1058, 1016, 892, 712,
696, 608, 498 NMR (CDCl ₃ ) δ: 1.37 (s, 9H), 3.10 (d, J = 6.2Hz, 2H), 4.52
(dd, J = 6.4Hz, J = 14.0Hz, 1H), 5.09 (dd, J = 12.2Hz, J = 14.0H
z, 2H), 5.31 (d, J = 7.8Hz, 1H), 7.10-7.28 (m, 4H), 7.35 (s,
5H)

Reference Example 34 ^t- butyl N-benzyloxycarbonyl-4-methoxycarbonyl-L-phenylalanate ^t -butyl N-
Benzyloxycarbonyl-4-trifluoromethanesulfonyloxy-L-phenylalanate 2.0 g, 1,
164 mg of 3-bis (diphenylphosphino) propane,
Palladium (II) acetate 89.1 mg and triethylamine 1.11 ml were dissolved in methanol 15.0 ml and dimethyl sulfoxide 20.0 ml, and carbon monoxide was bubbled for 5 minutes.
Furthermore, the mixture was stirred at 80 ° C. for 2 hours in a carbon monoxide atmosphere. Water was added to the reaction solution, and the mixture was extracted and extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure. The residue was purified by silica gel chromatography (hexane / ethyl acetate = 3/1) to give the title compound (1.52 g, 92.7%) as a colorless oil. IR ν max cm ^-1 : 3346, 2978, 2950, 1720, 1610, 1515, 1
452, 1434, 1367, 1351,1279, 1220, 1178, 1154, 110
7, 1056, 1020, 844, 751, 697 NMR (CDCl ₃ ) δ: 1.39 (s, 9H), 3.00-3.25 (m, 2H), 3.90 (s,
3H), 4.56 (dd, J = 7.0Hz, J = 13.0Hz, 1H), 5.09 (dd, J = 15.2H
z, J = 17.0Hz, 2H), 5.32 (d, J = 8.0Hz, 1H), 7.23 (d, J = 8.0H
z, 2H), 7.34 (s, 5H), 7.94 (d, J = 8.0Hz, 2H) Reference Example 35 N-benzyloxycarbonyl-4-methoxycarbonyl-L-phenylalanine ^t- butyl N-benzyloxycarbonyl-4- 0.55 g of methoxycarbonyl-L-phenylalanate was dissolved in 2.75 ml of trifluoroacetic acid and 2.75 ml of methylene chloride and stirred at room temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure and the toluene azeotrope was repeated several times. The residue was dissolved in a small amount of ethyl acetate and crystallized from petroleum ether to give the title compound 4
69 mg (98.7 mmol) were obtained as white crystals. Specific rotation [α] _D ²⁰ : + 58.6 ° (C = 0.1, CHC
l ₃ ) Melting point: 102-106 ° C. Elemental analysis value: C ₁₉ H ₁₉ NO ₆ calculated value: C, 63.89; H, 5.36; N, 3.92 Analysis value: C, 63.70; H, 5.47; N, 3.95

Reference Example 36 (S) -4- (N-benzyloxycarbonyl-4-methoxycarbonyl-L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t
Butyl ester (S) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester oxalate 0.76
g and N-benzyloxycarbonyl-4-methoxycarbonyl-L-phenylalanine 0.47 g were dissolved in methylene chloride 7.6 ml, and 1-ethyl-
3- (3-dimethylaminopropyl) carbodiimide 0.
44 g was added slowly over 5 minutes. After stirring at room temperature for 1 hour, methylene chloride was distilled off under reduced pressure. The residue was dissolved in ethyl acetate, washed with 5% aqueous potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate, the organic layer was dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was chromatographed on silica gel (hexane / ethyl acetate = 1 /
Purification in 2) gave 0.46 g (51.1%) of the title compound as a colorless oil. IR ν max cm ^-1 : 3322, 2980, 2952, 1719, 1649, 1525, 1
489, 1435, 1367, 1281,1152, 1104, 1020, 978, 846,
748, 699 NMR (CDCl ₃ ) δ: 1,45 (s, 9H), 2.61-2.72 (m, 1H), 2.72-3.
25 (m, 5H), 3.63 (s, 3H), 3.40-4.05 (m, 4H), 3.90 (s, 3H),
4.78-5.20 (m, 4H), 5.61 (d, J = 8.0Hz, 1H), 7.16-7.46 (m, 7
H), 7.95 (d, J = 8.0Hz, 2H)

Example 28 (S) -4- (4-amidinobenzoyl-L-4-methoxycarbonylphenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid (S) -4- ( N-benzyloxycarbonyl-4-methoxycarbonyl-L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid
0.46 g of ^t- butyl ester was dissolved in 4.6 ml of ethyl acetate, 0.19 g of 10% palladium-carbon was added, and the mixture was stirred under hydrogen for 3 hours. The catalyst was filtered off and the filtrate was concentrated to give a colorless oil. This product was dissolved in 3.0 ml of methylene chloride and 3.0 ml of trifluoroacetic acid and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and azeotropically distilled several times with toluene.
This product was dissolved in a mixed solvent of 8.1 ml of water and 4.0 ml of 1,4-dioxane, 0.33 g of sodium hydrogen carbonate was added at room temperature, and then 0.24 g of 4-amidinobenzoyl chloride was added at room temperature over 5 minutes. added. After stirring for 1 hour, 1N HCl
After adjusting the pH to 2-3 with water, the reaction solution was concentrated under reduced pressure, and the residue was subjected to CHP-20 column (H ₂ O → 5% CH ₃ CN / H ₂ O →
10% CH ₃ CN / H ₂ O → 15% CH ₃ CN / H ₂ O → 2
Purified with 0% CH ₃ CN / H ₂ O). 1NH
It is converted to the hydrochloride with Cl water and freeze-dried to give the title compound 0.2
6 g (53.0%) was obtained as a colorless amorphous powder. Specific rotation [α] _D ²⁰ : + 57.1 ° (C = 1.0%, H ₂
O) Elemental _{analysis: C 28 H 32 N 5 O} 9 · 2.5H 2 O Calculated: C, 50.72; H, 5.62 ; N, 10.56 analysis: C, 50.68; H, 5.50 ; N, 10.48

Reference Example 37 (S) -4- (N- ^t -butoxycarbonyl-4-formyl-
L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester (S) -4- (N- ^t -butoxycarbonyl-4-vinyl-L
-Phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester 2.
21 g, sodium metaperiodate 3.46 g, carbon tetrachloride 22.1 ml, acetonitrile 22.1 ml, water 33.2 m
The solution was dissolved in 1 l, and 16.4 mg of ruthenium (III) chloride nhydrate was added at room temperature and stirred for 1 hour. Ethyl acetate was added to the reaction solution for liquid separation and extraction, and the organic layer was concentrated under reduced pressure. The residue was chromatographed on silica gel (hexane / ethyl acetate = 1 /
Purification in 2) gave 1.14 g (51.4%) of the title compound as a colorless oil. IR ν max cm ^-1 : 2978, 2934, 1740, 1702, 1654, 1605, 1
489, 1436, 1366, 1247, 1228, 1157, 1047, 1016, 979,
848 NMR (CDCl ₃ ) δ: 1.40 (s, 9H), 1.45 (s, 9H), 2.66-3.27 (m,
6H), 3.42-3.93 (m, 6H), 4.05-4.36 (m, 1H), 4.72-4.90 (m,
1H), 4.98 (t, J = 5.0Hz, 1H), 5.35 (d, J = 9.0Hz, 1H), 7.34-
7.60 (m, 2H), 7.83 (d, J = 8.0Hz, 2H), 9.98 (s, 1H)

Example 29 (S) -4- (4-amidinobenzoyl-L-4-formylphenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid (S) -4- (N ^-T -butoxycarbonyl-4-formyl-
L-Phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester 0.22 g was dissolved in methylene chloride 1.1 ml and trifluoroacetic acid 1.1 ml and stirred at room temperature for 1 hour. . The reaction system was concentrated under reduced pressure and azeotropically distilled several times with toluene. This product 4.1 ml of water,
The mixture was dissolved in a mixed solvent of 2.0 ml of 1,4-dioxane, 0.16 g of sodium hydrogen carbonate was added at room temperature, and then 0.13 g of 4-amidinobenzoyl chloride was added at room temperature over 5 minutes. After stirring for 1 hour, with 1N HCl water, pH 2-3
The reaction mixture was concentrated under reduced pressure and the residue was concentrated on a CHP-20 column (H ₂ O → 5% CH ₃ CN / H ₂ O → 10% CH ₃ CN /
The product was purified by H ₂ O → 15% CH ₃ CN / H ₂ O) to obtain 0.15 g (64.9%) of the title compound as a white lyophilized product. Specific rotation [α] _D ²⁰ : + 28.6 ° (C = 1.0, H
₂ O) Elemental analysis value: C ₂₇ H ₂₉ N ₅ O ₈ 3.0H ₂ O Calculated value: C, 53.55; H, 5.83; N, 11.56 Analysis value: C, 53.61; H, 5.74; N, 11.68

Reference Example 38 (S) -4- (N- ^t -butoxycarbonyl-4-hydroxymethyl-L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester (S ) -4- (N- ^t -butoxycarbonyl-4-formyl-
L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester (0.65 g) was dissolved in methanol (13.0 ml), and sodium borohydride (21.9 mg) was added with stirring at 0 ° C. And stirred for 1 hour. Water was added to the reaction solution, and the mixture was separated and extracted with methylene chloride, and the organic layer was concentrated under reduced pressure. The residue was purified by silica gel chromatography (methylene chloride / ethyl acetate = 2/3) to give the title compound (0.56 g, 85.6%) as a colorless oil. IR ν max cm ^-1 : 3428, 2978, 2932, 1740, 1708, 1647, 1
489, 1437, 1366, 1248, 1156, 1048, 1015 NMR (CDCl ₃ ) δ: 1.44 (s, 9H), 1.45 (s, 9H), 2.18-2.40 (m,
1H), 2.52-3.25 (m, 6H), 3.63 (s, 3H), 3.40-4.02 (m, 3H),
4.30 (d, J = 16.0Hz, 1H), 4.63 (s, 2H), 4.68-4.94 (m, 2H),
5.23-5.50 (m, 1H), 7.10-7.40 (m, 4H)

Example 30 (S) -4- (4-amidinobenzoyl-L-4-hydroxymethylphenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid (S) -4- ( 0.26 g of N- ^t -butoxycarbonyl-4-hydroxymethyl-L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester was added to 2.0 ml of methylene chloride and 2.0 ml of trifluoroacetic acid. It was dissolved in 0 ml and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and azeotropically distilled with toluene several times. This product was dissolved in a mixed solvent of 4.8 ml of water and 2.4 ml of 1,4-dioxane, and 0.19 g of sodium hydrogen carbonate was added at room temperature.
0.15 g of amidinobenzoyl chloride was added over 5 minutes at room temperature. After stirring for 1 hour, p with 1N HCl water.
After adjusting to H2-3, the reaction solution was concentrated under reduced pressure and the residue was converted to CHP-2.
0 column (H ₂ O → 5% CH ₃ CN / H ₂ O → 10% CH ₃
After purification with CN / H ₂ O, 0.17 g (60.7) of the title compound was obtained.
%) Was obtained as a colorless lyophilized product. Specific rotation [α] _D ²⁰ : + 62.5 ° (C = 1.0, H
₂ O) Elemental analysis value: C ₂₇ H ₃₁ N ₅ O ₈・ 3.0H ₂ O Calculated value: C, 53.37; H, 6.14; N, 11.53 Analytical value: C, 53.50; H, 5.94; N, 11.46

Example 31 (S) -4- {4- (2-aminoethyl) benzoyl-
400 mg of O-methyl-L-tyrosyl} -3-methoxycarbonyl-2-oxopiperazine-1-acetic acid 4- (2-tert-butoxycarbonylaminoethyl) benzoic acid and 203 mg of 1-hydroxybenztriazole monohydrate were added to dichloromethane. Dissolve in 5 ml 1-
400 mg of ethyl-3- (3-dimethylaminopropyl) carbodiimide was added at room temperature and stirred for 1 hour. After adding 300 mg of triethylamine to this solution, (S) -4-O-methyl-L-tyrosyl-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid obtained in Reference Example 5 was obtained.
2 ml of a dichloromethane solution in which 785 mg of tert-butyl ester acetate was dissolved was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous sodium hydrogen carbonate solution and 5% aqueous potassium hydrogen sulfate solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The obtained crude product was dissolved in a mixture of 3 ml of dichloromethane and 3 ml of trifluoroacetic acid, allowed to stand at room temperature for 1 hour, and concentrated under reduced pressure. The residue was purified by MCI GEL CHP20P (Mitsubishi Kasei) (eluent H ₂ O → 15% CH ₃ CN) and freeze-dried to give 490 mg of the title compound as a colorless powder. Specific rotation [α] ^{_{D 20 68.4 ° (C = 0.45}} , H 2 O) Elemental _{analysis: C 28 H 34 N 4 O} 8 · 2H 2 O Calculated: C, 56.95; H, 6.48 ; N, 9.49. Found: C, 56.80; H, 6.61; N, 9.20.

Reference Example 39 (S) -4- (N-benzyloxycarbonyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-
2-Oxopiperazine-1-acetic acid (S) -3-methoxycarbonyl-2-oxopiperazine-1-acetic acid ^t -butyl ester 28.8 obtained in Reference Example 2
g, N-benzyloxycarbonyl-4-methoxyphenylalanine 23.0 g were dissolved in methylene chloride 202 ml, and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide 15.3 g was added. After stirring at room temperature for 2 hours, the reaction system was concentrated under reduced pressure, the residue was dissolved in ethyl acetate and washed with 5% aqueous potassium hydrogen sulfate solution and saturated aqueous sodium hydrogen carbonate solution. The organic layer was concentrated under reduced pressure to give crude (S) -4- (N
37.0 g of -benzyloxycarbonyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid ^t- butyl ester were obtained.
This product was dissolved in 74.0 ml of methylene chloride, and 74.0 ml of trifluoroacetic acid was added. After stirring at room temperature for 1 hour, the reaction system was concentrated under reduced pressure. The residue was azeotroped with toluene several times to give 39.12 g of the title compound as a colorless oil. IR ν max cm ^-1 : 3422, 1720, 1643, 1512, 1438, 1299, 1
246, 1177, 1027, 980, 823, 740, 696 NMR (CDCl ₃ ) δ: 2.30-2.90 (m, 6H),
2.90-3.83 (m, 7H), 3.71 (s, 3)
H), 3.90-4.15 (m, 1H), 4.50.
-4.75 (m, 1H), 4.80-4.95 (m,
1H), 4.96 (s, 2H), 6.81 (d, J
= 8.6 Hz, 2H), 7.10-7.40 (m, 7)
H)

Reference Example 40 (S) -4- (N-benzyloxycarbonyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-
2-oxopiperazine-1-acetic acid dimethylaminocarbonylmethyl ester (S) -4- (N-benzyloxycarbonyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxo obtained in Reference Example 39 Piperazine-1-acetic acid 2.
0 g, 83.0 mg of sodium iodide, 563 μl of N, N-dimethylaminocarbonylmethyl chloride were dissolved in 7.39 ml of N, N-dimethylformamide, and 772 μl of triethylamine was added with stirring at room temperature. After stirring for 6 hours, water was added to the reaction system for liquid separation and extraction with ethyl acetate, and the organic layer was concentrated under reduced pressure. Silica gel chromatography of the residue
The product was purified with (methylene chloride / methanol = 13/1) to obtain 1.57 g (68.0%) of the title compound as a colorless oily substance. IR ν max cm ^-1 : 2944, 1738, 1651, 1511, 1437, 1343, 1
246, 1202, 1028, 821,748, 697 NMR (CDCl ₃ ) δ: 2.47-2.68 (m, 1H), 2.76-3.23 (m, 5H), 2.
95 (s, 3H), 2.96 (s, 3H), 3.58-3.84 (m, 2H), 3.63 (s, 3H),
3.77 (s, 3H), 4.04 (d, J = 17.0Hz, 1H), 4.34 (d, J = 17.0Hz, 1
H), 4.72-4.89 (m, 3H), 4.92-5.06 (m, 1H), 5.09 (s, 2H),
5.54 (d, J = 10Hz, 1H), 6.80 (d, J = 8.4Hz, 2H), 7.08 (d, J = 8.4
Hz, 2H), 7.36 (s, 5H) By the same operation as in Reference Example 40, the following Reference Example No. 41-
43 compound was obtained as a colorless oil.

[0067]

[Table 8]

Example 32 (S) -4- (4-amidinobenzoyl-O-methyl-L)
-Tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid dimethylaminocarbonylmethyl ester hydrochloride (S) -4- (N-benzyloxycarbonyl-O-methyl-L-) obtained in Reference Example 40. Tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid dimethylaminocarbonylmethyl ester (1.50 g) was dissolved in methanol (15.0 ml), and 10% palladium-carbon (6.0 g) was added. After stirring for 1.5 hours at room temperature under a hydrogen atmosphere, the catalyst was filtered off and the filtrate was concentrated under reduced pressure.
570 mg of this product was added to 11.4 ml of water and 5.7 m of 1,4-dioxane.
dissolved in a mixed solvent of l, 0.29 g of sodium hydrogen carbonate
Was added at room temperature, then 0.38 g of 4-amidinobenzoyl chloride hydrochloride was added at room temperature over 5 minutes. After stirring for 1 hour, the pH was adjusted to 2-3 with 1N hydrochloric acid, and the reaction solution was concentrated under reduced pressure. The residue was subjected to CHP-20 column (H ₂ O → 5% CH _2
3 CN / H ₂ O → 10% CH ₃ CN / H ₂ O → 15% CH ₃
CN / H ₂ O), purified by LH-20 column (H ₂ O),
0.43 g (50.8%) of the title compound was obtained as a white lyophilized product. Specific rotation [α] _D ²⁰ : + 73.1 ° (C = 0.71, H ₂
O) Elemental analysis value: C ₃₁ H ₃₉ N ₆ O ₉ Cl.3.0H ₂ O (729.
183) as the calculated value: C, 51.06; H, 6.22; N, 11.53 Analytical value: C, 51.15; H, 5.94; N, 11.54 Example 3 using the compounds of Reference Example Nos. 41, 42 and 43
By the same operation as in Example 2, compounds of Examples No. 33-35 were obtained.

[0069]

[Table 9]

Example 36 (S) -4- {4- (2-aminoethyl) benzoyl-O-
Methyl-L-tyrosyl})-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid dimethylaminocarbonylmethyl ester hydrochloride (S) -4- (N-benzyloxycarbonyl-O-) obtained in Reference Example 40. Methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid dimethylaminocarbonylmethyl ester (1.50 g) was dissolved in methanol (15.0 ml) and 10% palladium-carbon (6.0 g) was added. After stirring for 1.5 hours at room temperature under a hydrogen atmosphere, the catalyst was filtered off and the filtrate was concentrated under reduced pressure.
570 mg of this product and 0.42 mg of 4- (2-benzyloxycarbonylaminoethyl) benzoic acid were added to 5.7 ml of methylene chloride.
And 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.29 g) was added over 5 minutes while cooling with water. After stirring for 2 hours, the reaction system was concentrated under reduced pressure, the residue was dissolved in ethyl acetate, washed with 5% aqueous potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate, and the organic layer was concentrated under reduced pressure. Silica gel chromatography of the residue
It was roughly purified with (methylene chloride / methanol = 13/1). This product was dissolved in 6.0 ml of methanol and 853 μl of 1N hydrochloric acid water, 180 mg of 10% palladium-carbon was added, and the mixture was stirred at room temperature under a hydrogen atmosphere for 40 minutes. The catalyst was filtered off, the filtrate was concentrated under reduced pressure, and the residue was collected on a CHP-20 column (H ₂
O → 5% CH ₃ CN / H ₂ O → 10% CH ₃ CN / H ₂ O →
15% CH ₃ CN / H ₂ O) to give the title compound 0.3.
9 g (69.6%) was obtained as a white lyophilized product. Specific rotation [α] _D ²⁰ : (+ 73.8 °, H ₂ O) Elemental analysis value: C ₃₂ H ₄₂ N ₅ O ₉ Cl.2.5H ₂ O (72
1.203) as the calculated value: C, 53.29; H, 6.57; N, 9.71 Analytical value: C, 53.27; H, 6.35; N, 9.81 Example 36 using the compounds of Reference Example Nos. 41, 42 and 43
Compounds of Example Nos. 37-39 were obtained by the same operation as described above.

[0071]

[Table 10]

Reference Example 44 (S) -4- (N-tert-butoxycarbonyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-
2-oxopiperazin-1-acetic acid (S) -4- (N-benzyloxycarbonyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid obtained in Reference Example 39 19
g was dissolved in 300 ml of methanol, 2.5 g of 10% palladium-carbon was added, and the mixture was stirred under hydrogen atmosphere for 1.5 hours. The catalyst was removed by filtration, the filtrate was concentrated under reduced pressure, and 200 ml of water and 50 ml of 1,4-dioxane were added to the residue to dissolve it. After adding 7.36 ml of triethylamine, dibutyl dicarbonate 8.4 was added under ice cooling.
g was added, pH was adjusted to 2 with 5% potassium hydrogen sulfate, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The extract was concentrated under reduced pressure to give the crude title compound (16.5 g) as a colorless powder. IR νmax cm ^-1 : 3400, 2975, 1735, 1710, 1665, 1510,
1440, 1365, 1300, 1245, 1210, 1170, 1030. NMR (CDCl ₃ ) δ: 1.42 (s, 9H), 2.30-4.20 (m, 10H), 3.64
(s, 3H), 3.76 (s, 3H), 4.76 (m, 1H), 4.99 (t, J = 5.2Hz, 1H),
5.12 (d, J = 8.4Hz, 1H), 6.82 (d, J = 8.4Hz, 2H), 7.09 (d, J =
8.4Hz, 2H).

Reference Example 45 (S) -4- (N- ^t -butoxycarbonyl-O-methyl-L)
- tyrosyl) -3-methoxycarbonylmethyl-2-oxo-piperazin-1-acetic acid 3- (phthalidylidene) was obtained ethyl ester Reference Example 44 (S) -4- (N- ^t butoxycarbonyl -O- methyl -L- Tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid 2.2 g,
1.52 g of potassium hydrogen carbonate was dissolved in 8.66 ml of N, N-dimethylformamide, and (Z) -3- (2-
Bromoethylidene) phthalide (1.55 g) was added. After stirring at room temperature for 2 hours, water was added to the reaction system, the mixture was extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure. The residue was chromatographed on silica gel (methylene chloride / ethyl acetate = 6/3.
Purification in 5) gave 1.82 g (63.2%) of the title compound as a colorless oil. IR ν max cm ^-1 : 2976, 2948, 1786, 1740, 1705, 1650,
1512, 1438, 1363, 1246, 1177, 978, 819, 764, 690,
635 NMR (CDCl ₃ ) δ: 1.43 (s, 9H), 2.37-2.58 (m, 1H), 2.76-3.
30 (m, 5H), 3.50-3.90 (m, 2H), 3.65 (s, 3H), 3.78 (s, 3H),
4.09 (dd, J = 17.0Hz, J = 24.6Hz, 2H), 4.64-4.82 (m, 1H),
4.92-5.11 (m, 3H), 5.36 (d, J = 8.0Hz, 1H), 5.75 (t, J = 7.4
Hz, 1H), 6.83 (d, J = 8.4Hz, 2H), 7.03-7.26 (m, 2H), 7.53-
7.83 (m, 3H), 7.92 (d, J = 8.2Hz, 1H) By the same operation as in Reference Example 45, the compounds of Reference Examples Nos. 46 and 47 were obtained as colorless oils.

[0074]

[Table 11]

Example 40 (S) -4- (4-amidinobenzoyl-O-methyl-L)
-Tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid (3-phthalidylidene) ethyl ester hydrochloride (S) -4- (N- ^t -butoxycarbonyl-4-methoxy-) obtained in Reference Example 45. L-phenylalanyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid
1.71 g of (3-phthalidylidene) ethyl ester was dissolved in 5.1 ml of methylene chloride and 5.1 ml of trifluoroacetic acid,
Stir at room temperature for 1.0 hour. The reaction system was concentrated under reduced pressure and azeotropically distilled several times with toluene. 850 mg of this product was added to H ₂ O 17.
It was dissolved in a mixed solvent of 0 ml and 8.5 ml of 1,4-dioxane, 0.53 g of sodium hydrogen carbonate was added at room temperature,
0.41 g of 4-amidinobenzoyl chloride hydrochloride was added over 5 minutes at room temperature. After stirring for 1 hour, the pH was adjusted to 2-3 with 1N hydrochloric acid, and the reaction solution was concentrated under reduced pressure. The residue was subjected to CHP-20 column (H ₂ O → 5% CH ₃ CN / H ₂ O → 1).
0% CH ₃ CN / H ₂ O → 15% CH ₃ CN / H ₂ O → 20
% CH ₃ CN / H ₂ O → 25% CH ₃ CN / H ₂ O → 30%
_{CH 3 CN / H 2 O)} , LH-20 column (MeOH), silica gel chromatography _{(CH 3 CN / H 2 O} = 5/1)
The title compound (0.27 g, 26.0%) was obtained as a white lyophilized product. Specific rotation [α] _D ²⁰ : + 51.5 ° (C = 0.77, H ₂
O) Elemental _{analysis: C 37 H 37 N 5 O} 10 · 0.7HCl · 0.3C
Calculated as F ₃ CO ₂ H · 3.5H ₂ O (834.510): C, 54.12; H, 5.43; N, 8.39 Analytical value: C, 54.11; H, 5.21; N, 8.64 Reference Example No. 46, The compounds of Example Nos. 41 and 42 were obtained by the same procedures as in Example 40 using the compound of 47.

[0076]

[Table 12]

Example 43 (S)-{4- (2-Aminoethyl) benzoyl-O-methyl-L-tyrosyl})-3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid (3-phthalidylidene) was obtained ethyl ester hydrochloride in reference example 45 (S) -4- (N- ^t butoxycarbonyl -O- methyl -L- tyrosyl) -3-methoxycarbonylmethyl-2-oxo-piperazine-1-acetic acid (3-phthalidylidene ) 1.71 g of ethyl ester was dissolved in 5.1 ml of methylene chloride and 5.1 ml of trifluoroacetic acid and stirred at room temperature for 1.0 hour. The reaction system was concentrated under reduced pressure and azeotropically distilled several times with toluene. 860 mg of this product and 531 μl of triethylamine were dissolved in 4.3 ml of methylene chloride, and 0.2 g of 1-hydroxybenztriazole, 0.37 g of 4- (2- ^t -butoxycarbonylaminoethyl) benzoic acid, 1-
(3-Dimethylaminopropyl) -3-ethylcarbodiimide (0.28 g) was added to a methylene chloride solution. After stirring for 1 hour, the reaction system was concentrated under reduced pressure, the residue was dissolved in ethyl acetate, washed with 5% potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate, and the organic layer was concentrated under reduced pressure. The residue was chromatographed on silica gel (methylene chloride / ethyl acetate = 1 /
It was roughly purified in 4). This product was dissolved in 2.64 ml of methylene chloride and 2.64 ml of trifluoroacetic acid. The reaction system was concentrated under reduced pressure, azeotroped several times with toluene, and converted into a hydrochloric acid salt with 1N aqueous hydrochloric acid, and the residue was converted into a CHP-20 column (H ₂ O → 5% C).
H ₃ CN / H ₂ O → 10% CH ₃ CN / H ₂ O → 15% CH
₃ CN / H ₂ O → 20% CH ₃ CN / H ₂ O → 25% CH ₃
CN / H ₂ O → 30% CH ₃ CN / H ₂ O), purified by LH-20 column (MeOH), 0.22 g (24.
7%) was obtained as a white lyophilized product. Specific rotation [α] _D ²⁰ : 49.3 ° (C = 0.31, H
₂ O) Elemental analysis value: C ₃₈ H ₄₀ N ₄ O ₁₀・ 0.7HCl ・ 0.3C
Calculated as F ₃ CO ₂ H.2.0H ₂ O (808.515): C, 57.34; H, 5.61; N, 6.93 Analytical value: C, 57.02; H, 5.35; N, 6.81 Reference Example No. 46, The compounds of Examples 44 and 45 were obtained by the same procedure as in Example 43 using the compound of 47.

[0078]

[Table 13]

[0079]

Formulation Example When the compound (I) of the present invention is used as a therapeutic agent for thrombosis, for example, it can be used by the following formulation. 1. Tablets (1) (S) -4- (4-amidinobenzoyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid 10 g (2) Lactose 90 g (3) Corn starch 29 g (4) Magnesium stearate 1 g 130 g Ingredients (1), (2) and 17 g of ingredient (3) were mixed and granulated with a paste made from 7 g of ingredient (3) into 5 g of ingredient (3). And component (4) are added and mixed, and the mixture is compressed by a compression tableting machine to produce 1000 tablets having a diameter of 7 mm and containing 10 mg of component (1) per tablet. 2. Injection (1) (S) -4- (4-amidinobenzoyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid 10 g (2) Sodium chloride 9 g Component (1) and ( 2) is dissolved in 1000 ml of distilled water,
Dispense 1 ml each into 1000 brown ampoules, replace with nitrogen gas, and seal. All steps are performed aseptically.

3. Tablet (1) (S) -4- (4-amidinobenzoyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid 10 g (2) Sodium chloride 9 g Component (1) And (2) are dissolved in 1000 ml of distilled water,
Dispense 1 ml each into 1000 brown ampoules, replace with nitrogen gas, and seal. All steps are performed aseptically. 4. Injection (1) (S) -4- (4-amidinobenzoyl-O-methyl-L-tyrosyl) -3-methoxycarbonylmethyl-2-oxopiperazine-1-acetic acid 10 g (2) Sodium chloride 9 g Component (1) ) And (2) are dissolved in 1000 ml of distilled water,
Dispense 1 ml each into 1000 brown ampoules, replace with nitrogen gas, and seal. All steps are performed aseptically.

Claims (13)

[Claims] 1. The formula: [In the formula, A represents an amidino group or an optionally substituted aminoethyl group, and R ¹⁰ represents a nitro group, a halogen atom, a lower alkenyl group, a lower alkynyl group, a hydroxymethyl group, a formyl group, a lower alkyloxycarbonyl group. And a group represented by the formula —OR ¹¹ (in the formula, R ¹¹ represents a hydrogen atom or an optionally substituted lower alkyl, lower alkenyl, lower alkynyl, lower alkanoyl, carbamoyl or methanesulfonyl group). R ¹² and R ¹³ each represent a hydrogen atom, a hydroxyl group, a lower alkoxy group or a halogen atom, X represents a hydroxyl group,
A p-hydroxyphenyl group or a carboxyl group which may be esterified or amidated is shown, Y is a carboxyl group which may be esterified or amidated, and n is 1 or 2. ] The compound or its salt represented by these. 2. R ¹⁰ is a hydroxyl group, a C _1-5 alkoxy group which may be substituted with a C _1-3 alkoxy group, a C _2-3 alkenyloxy group, a C _2-3 alkynyloxy group, a nitro group, Halogen atom, C _1-3 alkanoyloxy group, carbamoyloxy group optionally substituted by C _1-3 alkyl group, methanesulfonyloxy group, C _2-3 alkenyl group, C _2-3 alkynyl group, hydroxymethyl group , Formyl group and C
The compound according to claim 1, which is one selected from _1-3 alkyloxycarbonyl groups. 3. The compound according to claim 1, wherein R ¹⁰ is a hydroxyl group or a methoxy group. 4. R ¹⁰ is an ethoxy group, a methoxyethoxy group,
The compound according to claim 1, which is a propoxy group, a propargyloxy group, an allyloxy group, a fluorine atom or a chlorine atom. 5. The compound according to claim 1, wherein R ¹² and R ¹³ are both hydrogen atoms. 6. The compound according to claim 1, wherein n = 1. 7. The compound according to claim 1, wherein A is an amidino group, R ¹⁰ is a hydroxyl group or a methoxy group, R ¹² and R ¹³ are both hydrogen atoms, and n = 1. 8. A is an amidino group, R ¹⁰ is an ethoxy group, a propargyloxy group, an allyloxy group or a fluorine atom, R ¹² and R ¹³ are both hydrogen atoms, and n
The compound according to claim 1, wherein = 1. 9. The compound according to claim 1, wherein A is an aminoethyl group, R ¹⁰ is a hydroxyl group or a methoxy group, R ¹² and R ¹³ are both hydrogen atoms, and n = 1. 10. An aminoethyl group, R ¹⁰ is an ethoxy group, a propargyloxy group, an allyloxy group or a fluorine atom, R ¹² and R ¹³ are both hydrogen atoms, and n = 1. The compound according to 1. 11. X is --COOCH ₃ and Y is --COOH.
Alternatively, the compound according to claim 7, 8, 9 or 10, which is a group that can be converted into a carboxy group in vivo. 12. A cell adhesion inhibitor comprising the compound according to claim 1. 13. The formula: [In the formula, R ¹¹ represents a hydrogen atom, an optionally substituted lower alkyl group, a lower alkanoyl group, an optionally substituted carbamoyl group or a methanesulfonyl group, and R ¹² and R ¹³ represent a hydrogen atom and a hydroxyl group, respectively. Represents a lower alkoxy group or a halogen atom, X represents a hydroxyl group, p-hydroxyphenyl group or a carboxyl group which may be esterified or amidated, and Y represents a carboxyl group which may be esterified or amidated. Shown, n is 1
Or 2].