JPH09315975A - Sustained release agent of 2-piperazinone-1-acetic acid derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prepare a microcapsule, comprising an amorphous and water-soluble type 2-piperazinone-1-acetic acid derivative and a high-molecular polymer, having a high uptake ratio of the compound useful as a therapeutic agent, etc., for circulatory diseases and hardly causing the initial release and adverse effects. SOLUTION: This microcapsule comprises an amorphous and water-soluble type compound of the formula [A<1> and A<2> are each a proton accepting group or a group convertible into the proton accepting group; D is a hetero-atom and/or a spacer of a 2 to 6 atomic chain; R<1> is H or a hydrocarbon; R<2> is H or a residue after removing CH(NH2 )COOH from an α-amino acid; P is a hetero- atom and/or a spacer of a 1 to 10 atomic chain; Y is carboxyl; (n) is 0-8] or its salt and a high-molecular polymer (preferably a polymer of lactic acid- glycolic acid, etc.). (S)-4-(4-guanidinobenzoylamino)acetyl-3-[3-4- guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid hydrochloride, etc., are preferred as the compound of the formula or its salt.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an amorphous water-soluble 2-
The present invention relates to a microcapsule containing a piperazinone-1-acetic acid derivative and a method for producing the same.

[0002]

2. Description of the Related Art There are many reports on sustained-release microcapsules of various low-molecular-weight water-soluble drugs [JP-A-57 / 1982].
-118512, J. Pharm. Sci. , 75 , 750-
755 (1986)], many of them (1) have a large leakage of the drug to the external aqueous phase in the production process and have a low drug trap rate. (2) The resulting capsule is generally porous and has an initial stage. It has the drawback of large emission,
The current situation is that long-term sustained release has not been achieved. on the other hand,
In recent years, novel peptides or low molecular weight compounds having excellent cell adhesion regulation or inhibitory action have been found,
It is expected as a therapeutic agent for various diseases. For example, a compound having a GPIIb / IIIa antagonistic activity exhibits a remarkable platelet aggregation inhibitory activity and a cancer cell metastasis inhibitory activity, and is expected as a clinically useful drug [Sci., 233 , 4].
67-469 (1986); Sci., 238 , 1132.
-1134 (1987); Proc. Natl. Acad. Sc
i. USA, 87 , 2471-2475 (1990)].
As such an example, -Arg-Gly-Asp- (RGD)
A linear peptide or cyclic peptide containing the amino acid sequence of J. is known [J. Biol. Chem., 262 , 172
94-17298 (1987); JP-A-2-17479.
7 etc.]. In addition to peptides or peptide-based compounds, non-peptide compounds having antithrombotic activity are also disclosed
-264068 and European Patent Application Publication No. 50
5868. Compounds each having a 4- to 7-membered cyclic alkyleneimino such as a pyrrolidine ring,
And compounds having a piperidine ring and the like. Further, a compound having a piperidinone ring having a cell adhesion inhibitory action is disclosed in European Patent Application Publication No. 529
858. The above-mentioned known compounds cannot be said to be sufficient in terms of strength of activity, side effect (prolonging action of bleeding time, etc.), absorbability, stability, or duration of action, and therefore, they are not suitable for clinical application as pharmaceuticals. Further improvements were needed. Recently, new 2-piperazinone-1-acetic acid derivatives have been synthesized, and these compounds have a strong platelet aggregation inhibitory action based on their chemical structural characteristics, and at the same time, are highly safe. It has been found to have minor properties of side effects such as bleeding tendency. These compounds are found in many cardiovascular diseases (eg, thrombosis, transient cerebral ischemic attack, myocardial infarction, cerebral infarction,
Peripheral artery occlusion, arteriosclerosis obliterans, etc., tumor, inflammatory disease, or re-occlusion after coronary artery bypass surgery and prevention of re-occlusion of coronary artery after PTCA (percutaneous intracoronary angioplasty) It is expected to be applied for prevention of restenosis, secondary prevention after reopening of infarction. Particularly in patients with chronic diseases, long-term administration is required, and long-term sustained-release preparations are desired for clinical application thereof, but there is no report on sustained-release microcapsules for the above novel compounds.

[0003]

2-Piperazinone-1
-To provide a sustained-release microcapsule having a high uptake rate of an acetic acid derivative and a small initial release, and a method for producing the same.

[0004]

Means for Solving the Problems As a result of intensive studies for solving the above-mentioned problems, the present inventors have found that a microstructure containing an amorphous water-soluble 2-piperazinone-1-acetic acid derivative and a high molecular weight polymer. It was found that the capsules had a high uptake rate of the compound and a low initial release. As a result of further intensive research based on this, the present invention has been completed.

That is, the present invention provides an amorphous water-soluble formula (I):

Embedded image [In the formula, A ¹ and A ² are each a proton accepting group or a group capable of converting into a proton accepting group, and D is a hetero atom and / or an atomic chain of 2 to 6 optionally having a 5- or 6-membered ring. Spacer (however, the 5 or 6 membered ring is 2 depending on the bonding position)
Or -CH 3 converted with atomic chains), the R ¹ is a hydrogen atom or a hydrocarbon group, R ² represents a hydrogen atom or an α- amino acid
Represents a residue excluding (NH ₂ ) COOH, or R ¹ and R ²
May be bonded to form a 5- or 6-membered ring, and P is a spacer of 1 to 10 atom chain which may be through a heteroatom and / or a 5- or 6-membered ring (provided that the 5- or 6-membered ring is Represents a 2 or 3 atom chain depending on the bonding position), Y represents a carboxyl group which may be esterified or amidated, and n represents an integer of 0 to 8. ] 2-piperazinone-1-acetic acid derivative which is a compound or salt thereof represented by
The present invention provides a microcapsule containing [a compound (I) may be abbreviated below] and a polymer. Further, the present invention provides a dispersion obtained by dispersing an amorphous water-soluble compound represented by formula (I) or a salt thereof, a 2-piperazinone-1-acetic acid derivative, in an organic solvent solution of a polymer. Disperse in water phase to make s / o / w type emulsion,
Also provided are microcapsules obtained by subjecting to underwater drying. Furthermore, the present invention provides a dispersion obtained by dispersing an amorphous water-soluble compound represented by formula (I) or a salt thereof, a 2-piperazinone-1-acetic acid derivative, in an organic solvent solution of a polymer. Also provided is a method for producing microcapsules, which comprises dispersing in an aqueous phase to form an s / o / w emulsion, and subjecting the emulsion to water drying.

In the present specification, amino acids, peptides, protecting groups and the like are represented by abbreviations such as Ayupack-IBU Commission on Biochemical Nomenclature (IUPAC-IUB Commission).
on Biochemical Nomenclature) or an abbreviation commonly used in the art, and where amino acids may have optical isomers, the L form is shown unless otherwise specified. In the present specification, the microcapsule means a general term for microspheres, microcapsules, microparticles, nanoparticles, nanospheres, and nanocapsules. In the present specification, the s / o / w type emulsion means a solid / oil / water type (solid phase in oil-in-water) emulsion. The s phase means a solid phase, but is also meant to include fine particles and a gelled aqueous phase. In the present invention, it is possible to obtain sustained-release microcapsules containing a high content of the water-soluble compound (I) and having a small initial release.

The amorphous compound (I) used in the present invention is water-soluble. Compound (I) is water-soluble means that water is 100 m.
It means that the solubility in 1 at 20 ° C. is generally about 1 g / 100 ml or more. Preferably compound (I) is readily soluble in water. Compound 1 is water-soluble, meaning that it is water 1
Solubility in 00ml is generally about 5g / 100 at 20 ℃
It means more than ml. As described above, the compound (I) of the present invention has the formula (I) characterized in that (1) it has a proton accepting group at each of the 3 and 4 terminal substituents of the piperazine ring:

Embedded image [In the formula, A ¹ and A ² each represent a group capable of being converted into a proton accepting group or a proton donating group, and D is a hetero atom and / or a chain of 2 to 6 atoms which may have a 5- or 6-membered ring. (Provided that the 5- or 6-membered ring is converted into a 2- or 3-atom chain depending on the bonding position), R ¹ represents a hydrogen atom or a hydrocarbon group, and R ² represents a hydrogen atom or an α-amino acid to -CH. Indicates a residue excluding (NH ₂ ) COOH,
Or, R ¹ and R ² may be bonded to each other to form a 5- or 6-membered ring, and P is a hetero atom and / or a spacer having a chain of 1 to 10 which may be through a 5- or 6-membered ring ( However,
A 5- or 6-membered ring is represented by a 2- or 3-atom chain depending on the bonding position), Y is a carboxyl group which may be esterified or amidated, and n is an integer of 0 to 8]
Is a compound or a salt thereof. In particular, (2) A ¹
And A ² are each an optionally substituted amino, amidino or guanidino group or a group convertible to them, (3) A ¹ and A ² may be each substituted The compound according to (1) above, which is an oxadiazolyl or thiadiazolyl group, (4) A ¹ and A ² are 1) an amidino or guanidino group which may be substituted with C _2-8 alkoxycarbonyloxy, or 2) oxo, or The compound according to (1) above, which is an amino group optionally substituted with an oxadiazolyl group optionally substituted with C _1-4 alkyl optionally substituted with halogen, (5) A ¹ and A ² are each The compound according to (1) above, which is an unsubstituted amino, amidino or guanidino group, (6) D has the formula

Embedded image The compound according to (1) above, which is a group represented by:

[0008] (7) Compound of R ¹ is (1) wherein a hydrogen atom, substituted with a (8) R ² is a hydrogen atom or a C _1-4 phenyl group which may be substituted with an alkoxy group C The compound according to (1) above, which is a _1-4 alkyl group, (9) P is of the formula -Z-B- [wherein Z is

Embedded image (In these, any bond may be bonded to B) or a bond is

[Chemical 6] (A represents an integer of 0 to 2, b represents an integer of 0 to 2, c represents an integer of 1 to 5) or a bond (provided that both Z and B are bonds. except for). ] The compound described in (1) above, which is a group represented by

[Chemical 7] The compound according to the above (9), which is (any bond may be bonded to B),

(11) B

Embedded image Or - (CH _{2) d} - [d represents an integer of 1 to 4. ] The compound according to (9) above, (12) the compound according to (1) above, wherein Y is a carboxyl group or a C _1-6 alkoxy-carbonyl group, (13) above, wherein n is an integer of 1 to 4. (1) The compound described above, (14) The compound described in (1) above, wherein n is 2 or 3, (15) A ¹ and A ² may be substituted with 1) C _2-8 alkoxycarbonyloxy. An amidino or guanidino group, 2) oxo, or an amino group optionally substituted with an oxadiazolyl group optionally substituted with C _1-4 alkyl optionally substituted with halogen, or 3) oxo, or halogen An oxadiazolyl group optionally substituted with optionally substituted C _1-4 alkyl,

D is an expression

Embedded image A group represented by, R ¹ is a hydrogen atom, R ² is a hydrogen atom or C
_1-4 C _1-4 alkyl group substituted with an optionally substituted phenyl group with an alkoxy group, P is wherein -Z-B- (wherein, Z is

Embedded image (In the formula, b represents 0 or 1, and c represents an integer of 1 to 5.). ) Group,

Y is an expression

Embedded image A group represented by the formula (wherein R ⁷ is 1) hydroxyl group, 2) C _1-4 alkoxy-carbonyl or 5-methyl-2-oxo-1,
C _1-8 alkoxy or C _2-12 alkenyloxy group optionally substituted with 3-dioxolen-4-yl or 3) formula —OCH (R ^7a ) OCOR ⁸ (wherein R ^7a is a hydrogen atom or A C _1-6 alkyl group, R ⁸ represents a C _1-6 alkyl group or a C _5-7 cycloalkyloxy group). ) And n are integers of 1 to 4, the compound according to the above (1) or (9), (16) A ¹ and A ² are 1) amidino or guanidino group optionally substituted with methoxycarbonyloxy or 2 ) Amino group optionally substituted by 5-oxo-1,2,4-oxadiazol-3-yl or 5-trifluoromethyl-1,2,4-oxadiazol-3-yl,

D is

[Chemical 12] R ¹ is a hydrogen atom, R ² is a hydrogen atom or a p-methoxybenzyl group, and P is

Embedded image The above (1) in which Y is a carboxyl group and n is 2 or 3.
Preferred are the compounds described above and (17) the compounds described in (1) above, wherein A ¹ and A ² each are an unsubstituted amino, amidino or guanidino group and R ² is a hydrogen atom.

In the above formula (I), A ¹ and A ² each represent a proton accepting group or a group capable of being converted into a proton accepting group. In the above formula (I), the proton-accepting group represents a group that receives a proton from the partner, that is, a Bronsted base, and examples thereof include a group containing a nitrogen atom that can be positively charged. Furthermore, as a specific example of the proton accepting group,
Examples thereof include optionally substituted amino, amidino, or guanidino groups. The proton-accepting group is preferably an unsubstituted amino, amidino or guanidino group, or a C _1-4 alkyl-substituted secondary or tertiary amino (especially ethylamino), amidino or guanidino group.

The substituent of the optionally substituted amino, amidino or guanidino group is, for example, C
_1-6 alkyl groups (eg, methyl, ethyl, propyl,
Isopropyl, butyl, sec-butyl, tert-butyl,
Pentyl, hexyl etc.), C _2-6 alkenyl group (eg vinyl, allyl, isopropenyl, butenyl, isobutenyl, sec-butenyl etc.), C _2-6 alkynyl group (eg propargyl, ethynyl, butynyl, 1-hexynyl). Etc.), C _3-6 cycloalkyl groups (eg cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl etc.), C _6-14 aryl groups (eg phenyl, tolyl, xylyl, 1-naphthyl, 2-naphthyl, biphenyl, 2 -Indenyl, 2-anthryl and the like, especially a phenyl group), a C _7-16 aralkyl group (for example, benzyl, phenethyl, diphenylmethyl, triphenylmethyl, 1-
Naphthylmethyl, 2-naphthylmethyl, 2-diphenylethyl, 3-phenylpropyl, 4-phenylbutyl,
A chain or cyclic hydrocarbon group such as 5-phenylpentyl and the like, especially a benzyl group; carbamoyloxy optionally substituted by C _1-4 alkyl (eg, N, N-dimethylaminocarbonyloxy) and C _{2 -5} alkanoyloxy (for example, pivaloyloxy and the like) or a 5- or 6-membered heterocyclic group (for example, 2- or 3-thienyl, 2- or 3-)
Furyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, or 3-pyrrolidinyl, 2-, 4- or 5-oxazolyl, 2-, 4- or 5-thiazolyl, 3-, 4- or 5
-Pyrazolyl, 2-, 4- or 5-imidazolyl, 1,
2,3-triazolyl, 1,2,4-triazolyl, 1H
-Or a hetero atom selected from an oxygen atom, a sulfur atom, a nitrogen atom and the like in addition to a carbon atom such as 2H-tetrazolyl
5- to 4-membered ring groups such as 2-, 3- or 4-pyridyl, N-oxide-2-, 3- or 4-pyridyl,
2-, 4- or 5-pyrimidinyl, N-oxide-2
-, 4- or 5-pyrimidinyl, thiomorpholinyl, morpholinyl, piperidinyl, pyranyl, thiopyranyl,
1,4-oxazinyl, 1,4-thiazinyl, 1,3-thiazinyl, piperazinyl, triazinyl, 3- or 4-
Pyridazinyl, pyrazinyl, N-oxide-3- or 4
A 6-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom, nitrogen atom, etc. in addition to carbon atom such as pyridazinyl, preferably pyrrolidin-1-yl, morpholino etc.) C _1-4 alkyl group (eg,
Methyl etc.); C _2-8 alkoxycarbonyl (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, pentyloxycarbonyl, n-hexyloxycarbonyl, n-octyloxycarbonyl etc.); C _1-8 alkylaminocarbonyl (Eg, n-hexylaminocarbonyl, n-octylaminocarbonyl, etc.); C _2-8 alkoxycarbonyloxy (eg, methoxycarbonyloxy, ethoxycarbonyloxy, propoxycarbonyloxy, butoxycarbonyloxy, pentyloxycarbonyloxy, n- Hexyloxycarbonyloxy, n-octyloxycarbonyloxy and the like, preferably methoxycarbonyloxy and the like); 5- or 6-membered heterocyclic group (for example, 2
-Or 3-thienyl, 2- or 3-furyl, 1-, 2-
Or 3-pyrrolyl, 1-, 2- or 3-pyrrolidinyl,
2-, 4- or 5-oxazolyl, 2-, 4- or 5-
Thiazolyl, 3-, 4- or 5-pyrazolyl, 2, -4
-Or 5-imidazolyl, 1,2,3-triazolyl,
5-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom, nitrogen atom and the like in addition to carbon atom such as 1,2,4-triazolyl, 1H- or 2H-tetrazolyl,
For example, 2-, 3- or 4-pyridyl, N-oxide-2
-, 3- or 4-pyridyl, 2-, 4- or 5-pyrimidinyl, N-oxide-2-, 4- or 5-pyrimidinyl, thiomorpholinyl, morpholinyl, piperidinyl,
Pyranyl, thiopyranyl, 1,4-oxazinyl, 1,4
-Thiazinyl, 1,3-thiazinyl, piperazinyl, triazinyl, 3- or 4-pyridazinyl, pyrazinyl,
6-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom, nitrogen atom, etc. in addition to carbon atom such as N-oxide-3- or 4-pyridazinyl, preferably tetrahydrofuran-2-yl Etc.) etc. In addition, the substituent to the amino, amidino or guanidino group is 2
When one or more are present, they may be bonded to each other to form a 5- or 6-membered heterocycle (pyrrolidine, piperidine, morpholine, imidazoline, etc.).

The group which can be converted into a proton accepting group is
A group capable of converting into a proton-accepting group in the living body and accepting a physiologically active free proton is desirable. As such a group, an amidoxime group which may have a substituent on the oxygen atom [a specific example of the substituent is lower (C _1-4 ).
Alkyl (eg methyl, ethyl, propyl etc.), acyl (eg C _2-5 alkanoyl (eg pivaloyl etc.), benzoyl etc.), lower (C _1-4 ) alkoxy-carbonyl (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl). Etc.), lower (C _1-4 ) alkylthiocarbonyl (eg methylthiocarbonyl, ethylthiocarbonyl etc.), acyloxycarbonyl (eg C _2-5 alkanoyloxycarbonyl (eg pivaloyloxycarbonyl etc.), benzoyloxycarbonyl etc.) Optionally substituted C _6-12 aryloxycarbonyl (eg phenoxycarbonyl etc.) or C _7-14 aralkyloxycarbonyl (eg benzyloxycarbonyl etc.) (specific examples of the substituent include cyano, nitro , Amino, lower (C _1-4 ) alkoxy-carbonyl (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl etc.), lower (C _1-4 ) alkyl (eg methyl, ethyl, propyl etc.), lower (C
_1-4 ) Alkoxy (eg methoxy, ethoxy, propoxy etc.), mono- and di-lower (C _1-4 ) alkylamino (eg methylamino, ethylamino, propylamino, dimethylamino etc.), hydroxy, amido or lower (C _1-4 ) alkylthio (eg, methylthio, ethylthio, etc.) and the like, optionally substituted C
_6-12 aryl-carbonyl group (eg phenylcarbonyl etc.) (specific examples of the substituent are lower (C _1-4 ) alkyl (eg methyl, ethyl, propyl etc.), lower (C
_1-4 ) Alkenyl (eg vinyl, allyl etc.) or lower (C _1-4 ) alkynyl (eg ethynyl etc.) and the like, or an optionally substituted carbamoyl group (as specific examples of the substituent) Cyano, nitro, amino, lower (C _1-4 ) alkoxy-carbonyl (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl etc.), lower (C _1-4 ) alkyl (eg methyl, ethyl, propyl etc.), lower (C _1-4 ) Alkoxy (eg methoxy, ethoxy, propoxy etc.), mono- and di-lower (C _1-4 ) alkylamino (eg methylamino, ethylamino, propylamino, dimethylamino etc.), hydroxy, amido or lower (C _1-4 ) alkylthio (eg, methylthio, ethylthio, etc.) and the like], etc.], having a substituent Optionally an oxadiazolyl or thiadiazolyl group [wherein oxo as a substituent,
Thioxo, hydroxy, amino, mono- and di-lower (C
_1-4 ) Alkylamino (eg methylamino, ethylamino, propylamino, dimethylamino etc.), halogen (eg fluorine, bromine, chlorine etc.), cyano, azide, lower optionally substituted by halogen (C _1-4 ) Alkyl (eg trifluoromethyl etc.), lower (C _1-4 ) alkoxy (eg methoxy, ethoxy, propoxy etc.), lower (C _1-4 ) alkylthio (eg methylthio, ethylthio etc.), lower (C _1-4 ) Alkoxy-carbonyl (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl etc.), mono- or di-lower (C _1-4 ) alkylamino (eg methylamino, ethylamino, propylamino, dimethylamino etc.), lower (C _{1- 4} ) Alkylcarbamoyl (eg methylcarbamoyl, ethylcarbamoyl etc.), substituent C _6-12 aryl group which may have (eg phenyl etc.) (specific examples of the substituent are cyano, nitro, amino, lower (C _1-4 ) alkoxy-
Carbonyl (eg methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl etc.), lower (C _1-4 ) alkyl (eg methyl, ethyl, propyl etc.), lower (C _1-4 ) alkoxy (eg methoxy, ethoxy, propoxy etc.), Mono- and di-lower (C _1-4 ) alkylamino (eg methylamino, ethylamino, propylamino, dimethylamino etc.), hydroxy, amido or lower (C _1-4 ) alkylthio (eg methylthio, ethylthio etc.) and the like. Or a C _7-14 aralkyl group which may have a substituent (eg benzyl etc.) (specific examples of the substituent are cyano, nitro, amino, lower (C
_1-4 ) Alkoxy-carbonyl (eg, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, etc.), lower (C _1-4 ) alkyl (eg, methyl, ethyl, propyl, etc.), lower (C _1-4 ) alkoxy (eg, methoxy, Ethoxy, propoxy etc.), mono- and di-lower (C _1-4 ) alkylamino (eg methylamino, ethylamino, propylamino, dimethylamino etc.), hydroxy, amido or lower (C _1-4 ) alkylthio (eg methylthio, Ethylthio and the like) and the like) and the like] and the like, and among the oxadiazolyl or thiadiazolyl groups which may have a substituent, 1,2,4-oxa which may have a substituent, respectively. Diazole-3
Preference is given to the -yl or 1,2,4-thiadiazol-3-yl group. When the substituent is oxo or thioxo, it may be keto type or enol type. A C _6-12 aryloxycarbonyl or C _7-14 aralkyloxycarbonyl group which may have a substituent as a substituent of the above-mentioned amido oxime group or oxadiazolyl or thiadiazolyl group, an optionally substituted carbamoyl group, Among the C _6-12 aryl group which may have a substituent and the C _7-14 aralkyl group which may have a substituent, cyano, nitro, lower (C _1-4 ) alkoxy-carbonyl or Those substituted with lower (C _1-4 ) alkoxy are preferred. Among the above-mentioned C _6-12 aryl-carbonyl groups which may have a substituent as a substituent of the _amidoxime group, a hydrogen atom or lower (C _1-4 )
Those substituted with alkyl are preferred.

Specific examples of the group capable of being converted into a proton accepting group include 5-oxo-1,2,4-oxadiazol-3-yl group and 5-oxo-1,2,4-thiadiazole-3. -Yl group, 5-thioxo-1,2,4-oxadiazol-3-yl group, 5-thioxo-1,2,4-thiadiazol-3-yl group, 4-methyl-5-oxo-1, Two,
4-oxadiazol-3-yl group, 4-ethyl-5-
Oxo-1,2,4-oxadiazol-3-yl group, 4
-Propyl-5-oxo-1,2,4-oxadiazol-3-yl group, 1,2,4-oxadiazol-3-yl group, 5-ethoxycarbonyl-1,2,4-oxadi Azol-3-yl group, 5-carbamoyl-1,2,4-oxadiazol-3-yl group, 5-cyano-1,2,4-oxadiazol-3-yl group, 5-trifluoromethyl -1,2,4-oxadiazol-3-yl group, 5-phenyl-1,2,4-oxadiazol-3-yl group, 5-
Amino-1,2,4-oxadiazol-3-yl group, 5
-Propylamino-1,2,4-oxadiazole-3-
Group, 5-methylthio-1,2,4-oxadiazol-3-yl group, 5-azido-1,2,4-oxadiazol-3-yl group, amino (hydroxy) imino group, amino ( Methoxycarbonyloxy) imino group, amino (ethoxycarbonyloxy) imino group, amino (n-propyloxycarbonyloxy) imino group, amino (benzyloxycarbonyloxy) imino group, amino (p-
Nitrobenzyloxycarbonyloxy) imino group, amino (p-nitrophenyloxycarbonyloxy) imino group, amino (p-nitrobenzoyloxycarbonyloxy) imino group, amino (methoxy) imino group, amino (carbamoyloxy) imino group, Examples thereof include amino (methylcarbamoyloxy) imino group, amino (ethylcarbamoyloxy) imino group, amino (n-propylcarbamoyloxy) imino group and amino (n-butylcarbamoyloxy) imino group.

Among them, 5-oxo-1,2,4-oxadiazol-3-yl group, 5-oxo-1,2,4-thiadiazol-3-yl group, 5-ethoxycarbonyl-1,2,
4-oxadiazol-3-yl group, 5-cyano-1,
2,4-oxadiazol-3-yl group, 5-trifluoromethyl-1,2,4-oxadiazol-3-yl group, amino (methoxycarbonyloxy) imino group, amino (carbamoyloxy) imino group , An amino (methylcarbamoyloxy) imino group and an amino (ethylcarbamoyloxy) imino group are preferable. A ¹ and A ² are (1) C _2-8 alkoxycarbonyloxy-substituted amidino or guanidino group or (2) oxo, or halogen-substituted C
_An amino group which may be substituted with an oxadiazolyl group which may be substituted with _1-4 alkyl is preferable, and among them, an unsubstituted amino, amidino or guanidino group is preferable. Further, the compound (I) or a salt thereof in which A ¹ or A ² is a group capable of converting into a proton accepting group is advantageously used as an oral preparation.

In the above formula (I), D is a spacer of a chain of 2 to 6 atoms which may be through a hetero atom and / or a 5- or 6-membered ring (provided that the 5- or 6-membered ring is 2 or 6 depending on the bonding position). (Converted as 3 atom chain).

Embedded image In the present invention, it means that there is a space in which 2 to 6 atoms are connected in a straight line. In the above formula (I), a hetero atom represented by D and / or 5
Alternatively, examples of the hetero atom in the spacer (2 to 6 member chain) of the 2 to 6 atom chain which may be through a 6 membered ring include N, O, S and the like. The 5- or 6-membered ring may be a carbocycle or a heterocycle containing 1 to 4 heteroatoms selected from N, O and S, and may be a saturated ring or aromatic. It may be an unsaturated ring such as a ring. Examples of such a 5- or 6-membered ring include:

Embedded image And the like. Further, it is preferable that the 5- or 6-membered ring does not have a bond at an adjacent position on the ring. The 5- or 6-membered ring preferably has a bond at each of the second to third atom positions on the ring, but usually a saturated or unsaturated ring having a two to three atom chain (2 Or a three-membered chain), and a group having a chain of 2 to 6 as D as a whole is preferable. As the heteroatom intervening in the spacer represented by D, nitrogen is particularly preferable, and D bonded to a group represented by A ¹ such as an adjacent amidino group through an —NH— group is particularly preferable. The 5- or 6-membered ring may be directly bonded to an adjacent amidino group, or may be bonded to a group represented by A ¹ such as an amidino group via an —NH— group and further via a methylene chain. And may be bonded to a group represented by A ¹ such as an amidino group.

Further, D may be formed by directly connecting the adjacent carbonyl group and the above-mentioned 5- or 6-membered ring, bonding via a methylene chain, or bonding via a hetero atom. The methylene chain in D is

Embedded image [Wherein R ³ is hydrogen or a lower (C _1-4 ) alkyl group which may be substituted with an optionally substituted phenyl group, and R ⁴ is substituted with an optionally substituted phenyl group. Optionally a lower (C _1-4 ) alkyl group, an optionally substituted phenyl group or a benzyloxy group]. The substituent of the optionally substituted phenyl group as the substituent of the lower (C _1-4 ) alkyl group in R ³ or R ⁴ is a lower (C _1-4 ) alkyl group (eg methyl, ethyl, etc.) , Low grade (C _1-4 )
Examples thereof include alkoxy groups (eg, methoxy, ethoxy, etc.), halogens (eg, fluorine, chlorine, bromine, etc.), hydroxyl groups and the like. Examples of the lower (C _1-4 ) alkyl group for R ³ or R ⁴ include methyl, ethyl and the like.

A typical group represented by D is, for example, a group represented by the formula:

Embedded image The group represented by is preferable. Particularly, E is preferably a 5- or 6-membered ring. Further, h is preferably 0 or 1, m is preferably 0, 1 or 2, and k is preferably 0. Of the 5- or 6-membered ring represented by E, a benzene ring and a cyclohexane ring are preferable, and a benzene ring is particularly preferable. In the above formula (I)

Embedded image [In the formula, R ³ , R ⁴ and m have the same meanings as defined above] The group represented by arginine and homoarginine represents a substituent.

Embedded image (In these, any bond may be bonded to A ¹ )

In the above formula (I), R ¹ represents a hydrogen atom or a hydrocarbon group. The hydrocarbon represented by R ¹ is C
_1-6 alkyl groups (eg, methyl, ethyl, propyl,
Isopropyl, butyl, sec-butyl, tert-butyl,
Pentyl, hexyl etc.), C _2-6 alkenyl group (eg vinyl, allyl, isopropenyl, butenyl, isobutenyl, sec-butenyl etc.), C _2-6 alkynyl group (eg propargyl, ethynyl, butynyl, 1-hexynyl). Etc.), C _3-6 cycloalkyl groups (eg cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl etc.), C _6-14 aryl groups (eg phenyl, tolyl, xylyl, 1-naphthyl, 2-naphthyl, biphenyl, 2 -Indenyl, 2-anthryl and the like, especially a phenyl group), a _C7-16 aralkyl group (for example, benzyl, phenethyl, diphenylmethyl, triphenylmethyl, 1-naphthylmethyl, 2-naphthylmethyl, 2-diphenylethyl, 3- Phenylpropyl, 4-phenylbutyl, 5
-Phenyl pentyl, etc., and particularly a chain or cyclic hydrocarbon group such as benzyl group), and R ¹ is hydrogen,
Lower (C _1-4 ) alkyl (eg methyl, ethyl, propyl etc.) or benzyl (amongst others hydrogen) are preferred.

In the above formula (I), R ² represents a hydrogen atom or a residue obtained by removing —CH (NH ₂ ) COOH from α-amino acid. The group represented by R ² includes an α-amino acid
Any residue may be used except CH (NH ₂ ) COOH. Further, R ¹ and R ² may be bonded to each other to form a 5- or 6-membered ring.

Embedded image Rings such as are preferred. Usually, R ² is preferably an essential amino acid residue. In particular, R ² is hydrogen, a lower (C _1-4 ) alkyl group, a lower (C _1-4 ) alkyl group substituted with an optionally substituted phenyl group, a lower (C _1- ) group substituted with a hydroxyl group. ₄ ) A lower (C _1-4 ) alkyl group substituted with an alkyl group or a carbamoyl group is preferable.
As more specific groups, for example, hydrogen, methyl, isopropyl, sec-butyl, isobutyl, hydroxymethyl, benzyl, p-hydroxybenzyl, p-methoxybenzyl, carbamoylmethyl, carbamoylethyl and the like can be mentioned as representative examples. The substituent which may be substituted on the benzene ring of the optionally substituted phenyl group as the substituent of the lower (C _1-4 ) alkyl group in R ² is a lower (C _1-4 ) alkyl Groups (eg, methyl, ethyl, propyl, isopropyl, n-butyl,
sec-butyl group, etc.), lower (C _1-4 ) alkoxy group (eg, methoxy, ethoxy group, etc.), halogen (eg, chlorine, fluorine, bromine, etc.), hydroxyl group, etc., among which lower (C _{1- 4} ) Alkoxy groups are preferred. C _1-4 alkyl group is preferably substituted by also phenyl group substituted with a hydrogen atom or a C _1-4 alkoxy group as a group or atom represented by R ^2, among others p- hydroxybenzyl, p
-Methoxybenzyl or a hydrogen atom (among others, p-methoxybenzyl or a hydrogen atom, especially a hydrogen atom) is preferable.

In the above formula (I), n represents an integer of 0 to 8 (preferably 1 to 4, particularly preferably 2 or 3). In the above formula (I), P is a spacer of an atom chain of 1 to 10 which may be through a hetero atom and / or a 5- or 6-membered ring (provided that the 5- or 6-membered ring is 2 depending on the bonding position).
Or converted to a three-atom chain). Here, the spacer represented by P means a linear interval between (CH ₂ ) _n and A ² , and in the present invention, an interval in which 1 to 10 atoms are connected in a straight line is defined. It means having. As a spacer of 1 to 10 atom chain (1 to 10 member chain) optionally having a hetero atom represented by P and / or a 5- or 6-membered ring,

[Chemical 21] 1 to 4 (preferably 1 or 2) and / or 5 or 6 membered ring (wherein 5 or 6 membered ring is a carbocycle or hetero atom selected from N, O and S) It may be a hetero ring containing 1 to 4 of, or may be a saturated ring or an unsaturated ring such as an aromatic ring.

Embedded image Etc., and a benzene ring and a cyclohexane ring are preferable, and a benzene ring is particularly preferable. Examples of the hetero ring include 2- or 3-thienyl, 2- or 3-furyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, or 3
-Pyrrolidinyl, 2-, 4- or 5-oxazolyl, 2
-, 4- or 5-thiazolyl, 3-, 4- or 5-pyrazolyl, 2-, 4- or 5-imidazolyl, 1,2,3-
Triazolyl, 1,2,4-triazolyl, 1H- or 2
5-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom, nitrogen atom, etc. in addition to carbon atom such as H-tetrazolyl, for example, 2-, 3- or 4-pyridyl, N
-Oxide-2-, 3- or 4-pyridyl, 2-, 4-
Or 5-pyrimidinyl, N-oxide-2-, 4- or 5-pyrimidinyl, thiomorpholinyl, morpholinyl,
Carbons such as piperidinyl, pyranyl, thiopyranyl, 1,4-oxazinyl, 1,4-thiazinyl, 1,3-thiazinyl, piperazinyl, triazinyl, 3- or 4-pyridazinyl, pyrazinyl, N-oxide-3- or 4-pyridazinyl Examples thereof include 6-membered ring groups containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom, nitrogen atom and the like in addition to atoms, and piperazine or piperidine is preferable. And a divalent hydrocarbon group which may be interposed.

As a spacer having a hetero atom represented by P and / or an atom chain of 1 to 10 optionally via a 5- or 6-membered ring, more preferable examples include:

Embedded image Examples thereof include divalent hydrocarbon groups which may be 1 to 4 (preferably 1 or 2) selected from In the above formula (I), P is, for example, formula -Z-B- [wherein, Z is

Embedded image (These may be bonded to B by any bond) or a kind selected from the bonds, and B is

Embedded image (A and b represent an integer of 0 to 2 (preferably 0 or 1), c represents an integer of 1 to 5) or a bond (provided that Z and B are both bonds). ] And the like.

Of the above Z

[Chemical formula 26] Those represented by (any bond may be bonded to B) are preferable. Also, the above B is

Embedded image [Wherein, b represents an integer of 0 to 2 (preferably 0 or 1) and d represents an integer of 1 to 4]. Furthermore, the above B is

Embedded image Or - (CH _{2) d} - wherein, d is 1 to an integer of 4] is what is preferable.

The optionally amidated carboxyl group represented by Y is represented by the formula:

[Chemical 29] [Wherein, in NR ⁵ R ⁶ , R ⁵ and R ⁶ are the same or different and each represents hydrogen, a lower (C _1-6 ) alkyl group (eg, methyl, ethyl, propyl, butyl, hexyl, etc.), C _{2- 8}
Alkenyl groups (eg, allyl, 2-butenyl, 3-pentenyl, etc.), 5- or 6-membered heterocyclic groups (eg 2- or 3-)
Thienyl, 2- or 3-furyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, or 3-pyrrolidinyl, 2-4-
Or 5-oxazolyl, 2-, 4- or 5-thiazolyl, 3-, 4- or 5-pyrazolyl, 2-, 4- or 5
-Imidazolyl, 1,2,3-triazolyl, 1,2,4-
5-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom, nitrogen atom, etc. in addition to carbon atom such as triazolyl, 1H- or 2H-tetrazolyl, for example, 2
-, 3- or 4-pyridyl, N-oxide-2-, 3-
Or 4-pyridyl, 2-, 4- or 5-pyrimidinyl,
N-oxide-2-, 4- or 5-pyrimidinyl, thiomorpholinyl, morpholinyl, piperidinyl, pyranyl, thiopyranyl, 1,4-oxazinyl, 1,4-thiazinyl, 1,3-thiazinyl, piperazinyl, triazinyl, 3- or 4 6-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom, nitrogen atom, etc. in addition to carbon atom such as -pyridazinyl, pyrazinyl, N-oxide-3- or 4-pyridazinyl, preferably , Pyridyl) -substituted lower (C _1-4 ) alkyl (pyridylmethyl etc.) or C _6-12 aralkyl group (eg benzyl, phenethyl, phenylpropyl etc.), and the aryl group in the aralkyl group is not It may be substituted or may be substituted with 1 or 2 substituents. As the substituent,
Nitro, halogen (eg, chlorine, fluorine, bromine, etc.), lower (C _1-4 ) alkyl group (eg, methyl, ethyl etc.), lower (C _1-4 ) alkoxy group (eg, methoxy, ethoxy, propoxy etc.) And a group represented by

The optionally esterified carboxyl group represented by Y is represented by the formula:

Embedded image [Wherein R ⁷ is 1) a hydroxyl group, 2) an optionally substituted alkoxy, alkenyloxy or benzyloxy group [eg, a hydroxyl group, an optionally substituted amino (eg, amino,
N-lower (C _1-4 ) alkylamino (eg methylamino etc.), N, N-dilower (C _1-4 ) alkylamino (eg dimethylamino etc.), piperidino, morpholino etc.),
Halogen (eg chlorine, bromine, fluorine, etc.), lower (C
_1-6 ) alkoxy (eg, methoxy, ethoxy, etc.), lower (C _1-6 ) alkylthio (eg, methylthio, ethylthio, etc.), lower (C _1-4 ) alkoxy-carbonyl (eg, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, Isobutyloxycarbonyl etc.) or optionally substituted dioxolenyl (eg, 5-
Methyl-2-oxo-1,3-dioxolen-4-yl etc.) optionally substituted lower (C _1-8 ) alkoxy (eg methoxy, ethoxy, propoxy etc.), lower (C _2-12 ) alkenyl Oxy (eg vinyloxy, allyloxy, etc.) or benzyloxy group] or 3) formula —O
CH (R ^7a ) OCOR ⁸ [In the formula, R ^7a represents hydrogen, a linear or branched lower (C _1-6 ) alkyl group (eg, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl). , Sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, etc.) or a C _5-7 cycloalkyl group (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.), and R ⁸ is i) straight chain or branched A branched lower (C _1-6 ) alkyl group (eg, methyl, ethyl,
n-propyl, ylpropyl, n-butyl, isobutyl
sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, etc.), ii) lower (C _2-8 ) alkenyl group (eg, vinyl, propenyl, allyl, isopropenyl, etc.), iii) C _5-7 cyclo Alkyl groups (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.), iv) C _5-7
Lower (C _6-12 aryl group substituted by an optionally substituted C _6-12 aryl group such as cycloalkyl group (eg, cyclopentyl, cyclohexyl, cycloheptyl etc.) or phenyl group
_1-3 ) alkyl (eg, benzyl, p-chlorobenzyl,
Substituted with an _optionally substituted C _6-12 aryl group such as phenethyl, cyclopentylmethyl, cyclohexylmethyl), v) a C _5-7 cycloalkyl group (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.) or a phenyl group. A lower (C _2-3 ) alkenyl group (eg, those having an alkenyl moiety such as vinyl such as cinnamyl, propenyl, allyl, isopropenyl, etc.), vi) substituted with an optionally substituted phenyl group, etc. Good C _6-12 aryl groups (eg, phenyl, p-tolyl, naphthyl, etc.), vi
i) a linear or branched lower (C _1-6 ) alkoxy group (eg, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, t-butoxy, n-pentyloxy) , Isopentyloxy, neopentyloxy, etc.), viii) linear or branched lower (C _1-6 ) alkenyloxy group (eg, allyloxy, isobutenyloxy, etc.), ix) C _5-7 cycloalkyloxy group (eg , Cyclopentyloxy, cyclohexyloxy, cycloheptyloxy, etc.), x) C _5-7 cycloalkyl group (eg, cyclopentyl, cyclohexyl, cycloheptyl, etc.) or optionally substituted C _6-12 aryl group such as phenyl group Lower substituted with (C _1-3 )
Alkoxy groups (eg, those having an alkoxy moiety such as benzyloxy, phenethyloxy, cyclopentylmethyloxy, cyclohexylmethyloxy, methoxy, ethoxy, n-propoxy, isopropoxy, etc.), xi) C _5-7 cycloalkyl group (eg, Cyclopentyl, cyclohexyl, cycloheptyl, etc.) or a lower (C _2-3 ) alkenyloxy group substituted by an optionally substituted C _6-12 aryl group such as an optionally substituted phenyl group (eg, cinnamyloxy, etc. Vinyloxy, propenyloxy, allyloxy,
Those having an alkenyloxy moiety such as isopropenyloxy), xii) an optionally substituted C _6-12 aryloxy group such as an optionally substituted phenoxy group (eg, phenoxy, p-nitrophenoxy, naphthoxy, etc. ) Is shown. ]. ] The group etc. which are represented by these are preferable. In the above formula, when the substituent R ⁸ includes an optionally substituted C _6-12 aryl group, examples of the C _6-12 aryl group include phenyl and naphthyl (preferably phenyl). Substituents for the _6-12 aryl group include nitro, halogen (chlorine, fluorine, bromine, etc.), lower (C _1-4 ) alkyl (eg, methyl, ethyl, propyl, etc.), lower (C
_1-4 ) Examples thereof include alkoxy (eg, methoxy, ethoxy, propoxy, etc.), and among them, unsubstituted phenyl is preferably used. Y is preferably a carboxyl group which may be esterified with lower (C _1-4 ) alkyl (eg, carboxy, ethoxycarbonyl, etc.),
A carboxyl group is particularly preferable.

Among the compounds represented by the above formula (I) or salts thereof, the formula

[Chemical 31] [In the formula, A ¹ and A ² each may have an optionally substituted amino, amidino or guanidino group, an optionally substituted amidoxime group on the oxygen atom, or an optionally substituted group. oxadiazolyl or thiadiazolyl group, R ² is hydrogen, lower (C _1-4) alkyl group, substituted with also a phenyl group optionally being substituted lower (C _1-4) alkyl group, substituted by a hydroxyl group a lower ( A lower (C _1-4 ) alkyl group substituted with a C _1-4 ) alkyl group or a carbamoyl group, and P is

Embedded image 1 to 4 divalent hydrocarbons, Y is a carboxyl group which may be esterified or amidated, m is an integer of 0 to 2, and n is 0.
Is an integer of 8 to 8. ] The compound (Ia) represented by or its salt is preferable.

More preferably, as the compound (Ia) or a salt thereof, for example, A ¹ and A ² each may have an unsubstituted amino, amidino or guanidino group, or may each have a substituent 1,2, 4-oxadiazol-3-yl or 1,2,4-thiadiazol-3-yl group, R ² is p-hydroxybenzyl, p-methoxybenzyl or a hydrogen atom, and P is of the formula —ZB— [wherein , Z is

[Chemical 33] (These may be bonded to B by any bond) or a kind selected from the bonds, and B is

Embedded image (A and b represent an integer of 0 to 2 (preferably 0 or 1), c represents an integer of 1 to 5) or a bond (provided that Z and B are both bonds). .. ]
Is a carboxyl group which may be esterified or amidated, m is an integer of 0 to 2, and n is an integer of 1 to 4 or a salt thereof.

More preferably, as the above compound (Ia) or a salt thereof, for example, A ¹ and A ² each have an unsubstituted amino, amidino or guanidino group, or may each have a substituent 1,2, 4-oxadiazole-3-
Or a 1,2,4-thiadiazol-3-yl group, R ²
Is p-hydroxybenzyl, p-methoxybenzyl or a hydrogen atom, P is of the formula -Z-B- [wherein Z is

Embedded image And B is

Embedded image (B represents an integer of 0 to 2 (preferably 0 or 1)). ], A Y group is a carboxyl group which may be esterified or amidated, and m is 0 to 2
And n is an integer of 1 to 4 or a salt thereof.

As the compound (I) or a salt thereof, for example, A ¹ and A ² are (1) an amidino or guanidino group which may be substituted with C _2-8 alkoxycarbonyloxy, (2) oxo, or C _1-4 alkyl which may be substituted by halogen, amino group which may be substituted by oxadiazolyl group or (3) oxo, or C _1-4 alkyl which may be substituted by halogen An oxadiazolyl group which may be substituted with

Embedded image A group represented by, R ¹ is a hydrogen atom, R ² is a hydrogen atom or C
_1-4 C _1-4 alkyl group substituted with an optionally substituted phenyl group with an alkoxy group, P is wherein -Z-B- (wherein, Z is

Embedded image (In the formula, b represents 0 or 1, and c represents an integer of 1 to 5.). ) Group,

Y is an expression

Embedded image A group represented by the formula (wherein R ⁷ is 1) hydroxyl group, 2) C _1-4 alkoxy-carbonyl or 5-methyl-2-oxo-1,
C _1-8 alkoxy or C _2-12 alkenyloxy group optionally substituted with 3-dioxolen-4-yl or 3) formula —OCH (R ^7a ) OCOR ⁸ (wherein R ^7a is a hydrogen atom or A C _1-6 alkyl group, R ⁸ represents a C _1-6 alkyl group or a C _5-7 cycloalkyloxy group). And a compound in which n is an integer of 1 to 4 or a salt thereof is also preferable. As the compound (I) or a salt thereof, A ¹ and A ² are (1) an amidino or guanidino group which may be substituted with methoxycarbonyloxy, or (2) 5-oxo-1,2,4-oxadiazole. -3-
Or an amino group optionally substituted with 5-trifluoromethyl-1,2,4-oxadiazol-3-yl,

D is

Embedded image R ¹ is a hydrogen atom, R ² is a hydrogen atom or a p-methoxybenzyl group, and P is

Embedded image Most preferred are compounds where Y is a carboxyl group and n is 2 or 3 or salts thereof.

When the compound of the present invention is used as an oral preparation,
The optionally esterified carboxyl group as Y is, for example, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, propoxycarbonyl, pivaloyloxymethoxycarbonyl, 1- (cyclohexyloxycarbonyloxy) ethoxycarbonyl, 5- Methyl-2-oxo-1,3-dioxolene
4-ylmethoxycarbonyl, acetoxymethyloxycarbonyl, propionyloxymethoxycarbonyl,
n-Butyloxymethoxycarbonyl, isobutyloxymethoxycarbonyl, 1- (ethoxycarbonyloxy) ethoxycarbonyl, 1- (acetyloxy) ethoxycarbonyl, 1- (isobutyloxy) ethoxycarbonyl, 2- (isobutyloxycarbonyl) -2-
Propylidene ethoxycarbonyl, (3-phthalidylidene) ethoxycarbonyl and the like are preferable. The compound 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 salts of the compounds (I) and (Ia) of the present invention include inorganic salts such as hydrochloride, hydrobromide, sulfate, nitrate and phosphate, such as acetate, tartrate and citric acid. Organic salts such as salts, fumarates, maleates, toluene sulfonates, methane sulfonates, etc., metal salts such as sodium salts, potassium salts, calcium salts, aluminum salts, etc. such as triethylamine salts, guanidine salts, ammonium Examples thereof include pharmaceutically acceptable salts such as salts, hydrazine salts, quinine salts and cinchonine salts with bases. The compounds (I) and (Ia) and salts thereof may be hydrates or non-hydrates. Compound (I)
A ¹ and A ² are (1) C _1-6 alkyl group; C _2-6 alkenyl group; C _2-6 alkynyl group; C _3-6 cycloalkyl group; C _6-14 aryl group; C _{7- 16} aralkyl group; carbamoyloxy optionally substituted by C _1-4 alkyl, C _2-5
Alkanoyloxy, 5-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom and nitrogen atom in addition to carbon atom, or hetero atom selected from oxygen atom, sulfur atom and nitrogen atom in addition to carbon atom A C _1-4 alkyl group substituted by a 6-membered ring group containing 1 to 4 C; C _2-8 alkoxycarbonyl; C _1-8 alkylaminocarbonyl; C _2-8
Alkoxycarbonyloxy; 1 heteroatom selected from oxygen atom, sulfur atom and nitrogen atom in addition to carbon atom
A 5-membered ring group containing 4 to 4; or 1 hetero atom selected from an oxygen atom, a sulfur atom and a nitrogen atom in addition to the carbon atom.
To an amino, amidino or guanidino group which may each be substituted with a 6-membered ring group containing 4 to 4 (when two or more substituents to the amino, amidino or guanidino group are present, they are bonded to each other to form a 5- or 6-membered group). A heterocycle may be formed),

(2) C _1-4 alkyl; C _2-5 alkanoyl; benzoyl; C _1-4 alkoxy-carbonyl; C _1-4
Alkylthiocarbonyl; C _2-5 alkanoyloxycarbonyl; benzoyloxycarbonyl; cyano, nitro, amino, C _1-4 alkoxy-carbonyl, C _1-4 alkyl, C _1-4 alkoxy, mono or di C _1-4 alkylamino C _6-12 aryloxycarbonyl or C _7-14 aralkyloxycarbonyl, each of which may be substituted with hydroxy, amido or C _1-4 alkylthio;
A C _6-12 aryl-carbonyl group optionally substituted by C _1-4 alkyl, C _2-4 alkenyl or C _2-4 alkynyl; or cyano, nitro, amino, C _1-4 alkoxy-
Substituted on the oxygen atom with a carbamoyl group which may be substituted with carbonyl, C _1-4 alkyl, C _1-4 alkoxy, mono- or di-C _1-4 alkylamino, hydroxy, amido or C _1-4 alkylthio. Optionally amidoxime group or (3) oxo; thioxo; hydroxy; amino; mono- or di-C _1-4 alkylamino; halogen;
Cyano; azido; C _1-4 alkyl optionally substituted with halogen; C _1-4 alkoxy; C _1-4 alkylthio; C _1-4
Alkoxy-carbonyl; C _1-4 alkylcarbamoyl; cyano, nitro, amino, C _1-4 alkoxy-carbonyl, C _1-4 alkyl, C _1-4 alkoxy, mono- or di-C _1-4 alkylamino, hydroxy, amide Or C
C _6-12 aryl group optionally substituted by _1-4 alkylthio; or cyano, nitro, amino, C _1-4 alkoxy-carbonyl, C _1-4 alkyl, C _1-4 alkoxy, mono or di C _{1 -4} alkylamino, hydroxy, amido or C _7-14 optionally substituted by C _1-4 alkylthio
An oxadiazolyl or thiadiazolyl group each optionally substituted with an aralkyl group, D being a heteroatom and /
Or a 5- or 6-membered carbon ring or a 2- or 6-membered chain optionally having a 5- or 6-membered heterocycle containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom and nitrogen atom (provided that 5 or 6 A 6-membered carbon ring or a 5- or 6-membered heterocycle containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom and nitrogen atom is converted to a 2- or 3-membered chain depending on the bonding position), and R ¹ is hydrogen , C _1-6 alkyl group, C _2-6 alkenyl group, C _2-6 alkynyl group, C
_3-6 cycloalkyl group, C _6-14 aryl group or C _7-16 aralkyl group, R ² is hydrogen atom; C _1-4 alkyl group; C _1-4
Alkyl group, C _1-4 alkoxy, C _1-4 substituted by also phenyl group substituted by halogen or hydroxyl
Alkyl groups; C _1-4 alkyl group substituted by a hydroxyl group; or C _{1 -4} alkyl group substituted by a carbamoyl group, or R ¹ and R ² are combined to

Embedded image 1 to 10 member which P may be through a hetero atom and / or a 5 or 6 membered carbon ring or a 5 or 6 membered hetero ring containing 1 to 4 hetero atoms selected from oxygen atom, sulfur atom and nitrogen atom Chain (however, a 5- or 6-membered carbon ring or a 5- or 6-membered heterocycle containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom and nitrogen atom is converted to a 2- or 3-membered chain depending on the bonding position) ,

Y is an expression

Embedded image [In the formula, R ⁵ and R ⁶ are the same or different and each is a hydrogen atom; a C _1-6 alkyl group; a C _2-8 alkenyl group; a hetero atom selected from an oxygen atom, a sulfur atom and a nitrogen atom in addition to the carbon atom. 5-membered ring group containing 1 to 4 atoms or C _1-4 alkyl group substituted with 6-membered ring group containing 1 to 4 heteroatoms selected from oxygen atom, sulfur atom and nitrogen atom in addition to carbon atom ; Or nitro, halogen, C _1-4 alkyl or C
_1-4 represents a C _6-12 aralkyl group which may be substituted with alkoxy. ] Or a group represented by

Embedded image [In the formula, R ⁷ is 1) hydroxyl group, 2) hydroxyl group, amino, N—C _1-4 alkylamino, N, N
-Di C _1-4 alkylamino, piperidino, morpholino,
Halogen, C _1-6 alkoxy, C _1-6 alkylthio, C
_1-4 alkoxy-carbonyl or C _1-8 alkoxy, C _2-12 alkenyloxy or benzyloxy group optionally substituted by 5-methyl-2-oxo-1,3-dioxolen-4-yl or 3 ) Formula —OCH (R ^7a ) OCOR ⁸ [In the formula, R ^7a represents a hydrogen atom, a C _1-6 alkyl group, or a C _5-7 cycloalkyl group, and R ⁸ is i) a C _1-6 alkyl group, ii) C _2-8 alkenyl group, iii) C _5-7 cycloalkyl group, iv) C _5-7 cycloalkyl group or substituted with nitro, halogen, C _1-4 alkyl or C _1-4 alkoxy C _1-3 alkyl substituted with a good C _6-12 aryl group, v) C _5-7 cycloalkyl group or C optionally substituted with nitro, halogen, C _1-4 alkyl or C _2-4 alkoxy _6-12 C _2-3 alkenyl group substituted with an aryl group, vi) nitro, halogen , A C _6-12 aryl group optionally substituted by C _1-4 alkyl or C _1-4 alkoxy, vii) C _1-6
Alkoxy group, viii) C _1-6 alkenyloxy group, ix) C
_5-7 cycloalkyloxy group, x) C _5-7 cycloalkyl group or nitro, halogen, C _1-4 alkyl or C _1-4
A C _1-3 alkoxy group substituted by an optionally substituted C _6-12 aryl group, xi) a C _5-7 cycloalkyl group or nitro, halogen, C _1-4 alkyl or C
_1-4 C _2-3 alkenyloxy group substituted by C _6-12 aryl group optionally substituted by alkoxy, xii) nitro,
It represents a C _6-12 aryloxy group which may be substituted with halogen, C _1-4 alkyl or C _1-4 alkoxy. ]
And n is an integer of 0 to 8. ) Compounds are included.

Specific preferred compounds include 4-
(4-Amidinobenzoyl) aminoacetyl-3- [3
-(4-Amidinobenzoyl) aminopropyl] -2-
Oxopiperazine-1-acetic acid, 4- (4-amidinobenzoyl) aminoacetyl-3- [4- (4-amidinobenzoyl) aminobutyl] -2-oxopiperazine-1
-Acetic acid, 4- (4-amidinobenzoyl) aminoacetyl-3- [2- (4-amidinobenzoyl) aminoethyl] -2-oxopiperazine-1-acetic acid, 4- (4-amidinobenzoyl) aminoacetyl-3 -[2- (4-
Amidinophenylaminocarbonyl) ethyl] -2-oxopiperazine-1-acetic acid, 4- (4-amidinobenzoyl) aminoacetyl-3- [3- (4-amidinophenylaminocarbonyl) propyl] -2-oxopiperazine-1 -Acetic acid, 4- (4-amidinobenzoyl) aminoacetyl-3- [4- (4-amidinophenylaminocarbonyl) butyl] -2-oxopiperazine-1-acetic acid, 4- (4-guanidinobenzoyl) aminoacetyl- 3- [2- (4-guanidinobenzoylamino) ethyl] -2-oxopiperazine-1-acetic acid, 4- (4-guanidinobenzoyl) aminoacetyl-3- [3- (4
-Guanidinobenzoylamino) propyl] -2-oxopiperazine-1-acetic acid, 4- (4-guanidinobenzoyl) aminoacetyl-3- [4- (4-guanidinobenzoylamino) butyl] -2-oxopiperazine-1
-Acetic acid), 4- (4-amidinobenzoylamino) acetyl-3- [2- (4-guanidinobenzoylamino)
Ethyl] -2-oxopiperazine-1-acetic acid, 4- (4
-Amidinobenzoylamino) acetyl-3- [4-
(4-guanidinobenzoylamino) butyl] -2-oxopiperazine-1-acetic acid, 4- [4- (2-aminoethyl) benzoylamino] acetyl-3- [2- (4-
Amidinobenzoylamino) ethyl] -2-oxopiperazine-1-acetic acid, 4- [4- (2-aminoethyl) benzoylamino] acetyl-3- [3- (4-amidinobenzoylamino) propyl] -2-oxo Piperazine-1-acetic acid, 4- [4- (2-aminoethyl) benzoylamino] acetyl-3- [4- (4-amidinobenzoylamino) butyl] -2-oxopiperazine-1-acetic acid, 4- (4 -Amidinobenzoylamino) acetyl-
3- [3- (4-guanidinobutanoylamino)] propyl-2-oxopiperazine-1-acetic acid, 4- (4-amidinobenzoylamino) acetyl-3- [3- (4-
Guanidinobutanoylamino)] propyl-2-oxopiperazine-1-acetic acid, (S, S)-[3- [3- (4
-Guanidinobenzoylamino) propyl] -4- [3
-(4-Methoxyphenyl) -2- [4- (5-trifluoromethyl- [1,2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxopiperazin-1-yl] Acetic acid, (S, S)-[4-
[3- (4-Methoxyphenyl) -2- [4- (5-trifluoromethyl [1,2,4] oxadiazole-3-
Ilamino) benzoylamino] propionyl] -2-
Oxo-3- [3- [4- (5-trifluoromethyl [1,2,4] oxadiazol-3-ylaminobenzoylamino] propyl] piperazin-1-yl] acetic acid,
(S, S)-[4- [3- (4-methoxyphenyl)-
2- [4- (5-oxo-4,5-dihydro [1,2,
4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo-3- [4- (5-
Oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoylamino] propyl] piperazin-1-yl] acetic acid or (S, S) -4-
[2- (4-guanidinobenzoyl) amino-3- (4
-Methoxyphenyl) propionyl] -3- [3- (4
-Guanidinobenzoyl) aminopropyl] -2-oxopiperazine-1-acetic acid or a salt thereof and the like.

Among them, (S) -4- (4-amidinobenzoyl) aminoacetyl-3- {3- (4-amidinobenzoyl) amino} propyl-2-oxopiperazine-1
-Acetic acid (the trifluoroacetic acid salt of this compound may be hereinafter referred to as Compound B), (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] Propyl-2-oxopiperazine-1-acetic acid (the hydrochloride of this compound may be hereinafter referred to as compound A), (S) -4- (4-amidinobenzoylamino) acetyl-3- [2- (4 -Guanidinobenzoylamino)] ethyl-2-oxopiperazine-1-acetic acid (the hydrochloride salt of this compound is referred to as compound D
(S) -4- [4- (2-aminoethyl) benzoylamino] acetyl-3- [3- (4-amidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid ( The trifluoroacetic acid salt of this compound may be referred to as compound C hereinafter.) (S) -4- (4-amidinobenzoylamino) acetyl-3- [3- (4-guanidinobutanoylamino)]
Propyl-2-oxopiperazine-1-acetic acid (The hydrochloride of this compound may be hereinafter referred to as compound E) (S, S)-[3- [3- (4-guanidinobenzoylamino) propyl] -4 -[3- (4-methoxyphenyl) -2- [4- (5-trifluoromethyl- [1,2,
4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxopiperazin-1-yl] acetic acid, (S, S)-[4- [3- (4-methoxyphenyl) -2- [4] -(5-trifluoromethyl [1,
2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo-3- [3-
[4- (5-trifluoromethyl [1,2,4] oxadiazol-3-ylaminobenzoylamino] propyl] piperazin-1-yl] acetic acid, (S, S)-[4-
[3- (4-Methoxyphenyl) -2- [4- (5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoylamino] propionyl]
-2-oxo-3- [4- (5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino)
Benzoylamino] propyl] piperazin-1-yl]
Acetic acid or (S, S) -4- [2- (4-guanidinobenzoyl) amino-3- (4-methoxyphenyl) propionyl] -3- [3- (4-guanidinobenzoyl) aminopropyl] -2-oxo Piperazine-1-acetic acid or a salt thereof is preferable, and particularly (S) -4- (4
-Amidinobenzoylamino) acetyl-3- {3-
(4-Amidinobenzoyl) amino} propyl-2-oxopiperazine-1-acetic acid, (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2 -Oxopiperazine-1-acetic acid, (S) -4- (4-amidinobenzoylamino) acetyl-3- [2- (4-guanidinobenzoylamino)]
Ethyl-2-oxopiperazine-1-acetic acid or (S) -4- [4- (2-aminoethyl) benzoylamino] acetyl-3- [3- (4-amidinobenzoylamino)] propyl-2-oxopiperazine -1-Acetic acid or a salt thereof is preferable, and particularly (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-
Guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid or a salt thereof is preferable, and more preferably (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino). )] Propyl-2-oxopiperazine-1-acetic acid or acid addition salts thereof, most preferably (S) -4-
(4-guanidinobenzoylamino) acetyl-3-
[3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid hydrochloride and the like.
Further, 4- (4-guanidinobenzoyl) aminoacetyl-3- [3- (4-guanidinobenzoylamino) propyl] -2-oxopiperazine-1-acetic acid or a salt thereof is also preferable.

The compounds (I) and (Ia) of the present invention can be produced, for example, by the following method or a method analogous thereto. formula

Embedded image [Wherein each symbol has the same meaning as defined above] (I
I) or a reactive derivative thereof or a salt thereof, and a formula

Embedded image [Wherein each symbol has the same meaning as defined above] (II
It can be produced by reacting I) or a salt thereof. Examples of the salt of compound (II) or compound (III) include inorganic salts such as hydrochloride, hydrobromide, sulfate, nitrate and phosphate, such as acetate, tartrate, citrate and fumaric acid. Organic salts such as salts, maleates, toluenesulfonates and methanesulfonates, metal salts such as sodium salts, potassium salts, calcium salts and aluminum salts such as triethylamine salts, guanidine salts, ammonium salts and hydrazine salts. , Quinine salts, cinchonine salts and the like, and pharmaceutically acceptable salts such as salts with bases. As the reactive derivative of the compound (II), for example,
formula,

Embedded image [In the formula, A ¹ has the same meaning as described above, and W represents halogen (preferably a base). ] The compound (II) represented by or corresponding acid anhydride, azide, active ester [alcohol (for example, pentachlorophenol, 2,4,5-trichlorophenol, 2,4-dinitrophenol, cyanomethyl alcohol, para. Ester with nitrophenol, N-hydroxy-5-norbornene-2,3-dicarboximide, N-hydroxysuccinimide, N-hydroxyphthalimide, N-hydroxybenztriazole), and the like.

As the condensation reaction for producing the compounds (I) and (Ia) of the present invention, an amide bond-forming reaction in a conventional peptide, for example, a method using an active ester or a mixed acid anhydride or acid chloride is used. It can be carried out. For example, in the condensation reaction of compound (II) and (III), compound (II) and 2,4,5-trichlorophenol, pentachlorophenol, 2-nitrophenol,
Or phenols such as 4-nitrophenol or N-hydroxy such as N-hydroxysuccinimide, N-hydroxy-5-norbornene-endo-2,3-dicarboximide, 1-hydroxybenztriazole and N-hydroxypiperidine It can also be carried out by condensing the compound in the presence of a condensing reagent such as dicyclohexylcarbodiimide, converting it into an active ester form, and then condensing. Moreover, after obtaining the mixed acid anhydride by reacting the compound (II) with isobutyl chloroformate,
It can also be condensed. Further, the condensation reaction of the compound (II) or its reactive derivative with the compound (III) is carried out by using a peptide condensation reagent such as dicyclohexylcarbodiimide, N, N-carbonyldiimidazole, diphenylphosphoric acid azide or diethyl cyanophosphate alone. Can also be condensed. In the condensation reaction, the amidino group, guanidino group, or amino group present in compound (II), its reactive derivative or compound (III) is an inorganic acid (eg, hydrogen chloride, sulfuric acid, nitric acid, hydrogen bromide, etc.). It is desirable that it exists as a salt of or is protected with a tert-butoxycarbonyl group or a benzyloxycarbonyl group. In the condensation reaction, the carboxyl group present in compound (II), its reactive derivative or compound (III) exists as a salt of an inorganic acid (eg, hydrogen chloride, sulfuric acid, nitric acid, hydrogen bromide, etc.). Alternatively, it is preferably protected with a methyl, ethyl, benzyl or tert-butyl group. In the condensation reaction, the hydroxyl group present in compound (II), its reactive derivative or compound (III) is an inorganic acid (eg,
It is preferably present as a salt of hydrogen chloride, sulfuric acid, nitric acid, hydrogen bromide, etc.) or protected with a benzyl or tert-butyl group.

In any of the above cases, the condensation reaction is preferably an organic base (eg, triethylamine, N-methylpiperidine, 4-N, N-dimethylaminopyridine, etc.) or an inorganic base (sodium hydrogen carbonate, sodium carbonate, carbonic acid). The reaction can be accelerated by the addition of potassium, etc.). The reaction temperature is usually −20 to + 50 ° C., preferably around 0 ° C. to + 30 ° C., and the reaction time depends on the type of solvent (in the case of a mixed solvent, the mixing ratio thereof), the reaction temperature, etc. It is 1 minute to 72 hours, preferably about 15 minutes to 5 hours. Examples of commonly used solvents include water, dioxane, tetrahydrofuran, acetonitrile, pyridine,
Examples thereof include N, N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone, chloroform and methylene chloride, which may be used alone or as a mixed solvent. When a carboxyl group-protecting group (benzyl group or tert-butyl group which is a carboxyl group-protecting group of Y in the formula (I)) is present in the product of the final method, it is removed by a method known per se. be able to. For example, a compound having a benzyl ester group can be hydrogenated in the presence of a noble metal catalyst such as palladium or platinum to obtain a carboxylic acid derivative, and a compound having a tert-butyl ester group can be trifluoroacetic acid or a chloride. The carboxylic acid derivative can be obtained by treatment with an acid such as hydrogen. In the case where an amino-protecting group (tert-butoxycarbonyl group or benzyloxycarbonyl group, which is the amino-protecting group of X'shown in the following reaction formula) is present in the product of the final method, the method is known per se. Can be removed. For example, a compound having a tert-butoxycarbonyl group 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. Further, the compound having a benzyloxycarbonyl group can be removed by catalytic reduction using a metal such as platinum, palladium, Raney nickel or a mixture thereof with an arbitrary carrier as a catalyst. The salt of compound (I) can be obtained by the reaction itself for producing compound (I), but if necessary, an acid, an alkali or a base may be added to produce the salt of compound (I) as described above. it can.

The compound (I) obtained by the above-mentioned production method is separated from the reaction mixture by a conventional separation / production operation such as extraction, concentration, neutralization, filtration, recrystallization, column chromatography, thin layer chromatography and the like. Can be separated. Compound (I) can exist in at least two stereoisomers. Any of these individual isomers and mixtures thereof are within the scope of the invention. Moreover, these isomers can also be manufactured individually. A single optical isomer of compound (I) can be obtained by performing the reaction described above using a single isomer of compound (III). When the product is a mixture of two or more isomers, it is separated by a conventional separation method such as an optically active acid (eg, camphorsulfonic acid, tartaric acid, etc.), an optically active base (eg, cinchonine, cinchonidine, quinine, quinidine). , Α-methylbenzylamine, etc.) and various isomers can be separated by various methods such as chromatography, fractional recrystallization and the like.

The starting compounds (II) and (III) of the present invention are compounds known per se or can be produced according to a method known per se. Compound (III) can be produced according to a method known per se, for example, a method represented by the following reaction formula.

Embedded image In the above reaction, R is an amino-protecting group and represents a benzyloxycarbonyl group or a tert-butoxycarbonyl group. X'is a protected amino group (a benzyloxycarbonyl group, a tert-butoxycarbonyl group or the like is used as a protecting group), a protected carboxyl group (a methyl, ethyl, benzyl, tert-butyl group or the like as a protecting group) Used), a protected hydroxyl group (a benzyl group, a tert-butyl group or the like is used as a protecting group) and a protected mercapto group (a benzyl group, a trityl group or the like is used as a protecting group). Y'represents a protected carboxyl group (benzyl or tert-butyl group as a protecting group).

The method for producing the compound (III) represented by the above reaction scheme will be described in more detail. (IV) and (V)
The reaction to give (VI) is a normal alkylation reaction for amino groups, and the reaction between compound (IV) and compound (V)
In the presence of a base (eg, an inorganic base such as sodium carbonate, potassium carbonate, potassium hydrogen carbonate, cesium fluoride, or an organic base such as triethylamine, pyridine, 4-N, N-dimethylaminopyridine, etc.). It can be produced by reacting at a temperature of about 100 ° C. for about 15 minutes to 5 hours. As a reaction solvent, acetonitrile,
N, N-dimethylformamide, tetrahydrofuran,
Examples of the organic solvent include toluene and methylene chloride. Then, the production reaction of compound (VIII) by condensation of compound (VI) and compound (VII) is an ordinary peptide bond formation reaction, and should be performed under the same reaction conditions as the condensation reaction of compound (II) and compound (III). You can The cyclization reaction from compound (VIII) to compound (IX) is an acid-catalyzed cyclization reaction, and, for example, p-toluenesulfonic acid, camphorsulfonic acid, methanesulfonic acid or the like can be used as a catalyst. The reaction can usually be carried out in a solvent such as toluene, benzene, ethyl acetate or 1,2-dichloroethane at a reaction temperature of 0 to 100 ° C, preferably 30 to 80 ° C to produce the compound (IX).

Then, the reduction reaction from compound (IX) to compound (X) is carried out, for example, by catalytic reduction using a metal such as platinum, palladium, Raney nickel or the like and a mixture thereof with any carrier, for example sodium borohydride. Can be carried out by reduction with a metal hydride compound such as The reaction is usually performed in an organic solvent (eg, methanol, ethanol, dioxane, ethyl acetate, etc.), and the reaction temperature is generally −
20-100 degreeC is preferable. This reaction can be carried out under normal pressure and under pressure. When R is a benzyloxycarbonyl group, the deprotection reaction of R also simultaneously proceeds by the catalytic reduction reaction to obtain the compound (XI). (X) to (X
The deprotection reaction of (I) and (XVI) to (III) is a general reaction for removing an amino protecting group, and when R is a benzyloxycarbonyl group, a metal such as platinum, palladium, Raney nickel, or the like and any carrier It can be removed by catalytic reduction using a mixture of When R is a tert-butoxycarbonyl group, methanol, ethanol,
It can be easily removed with an acid such as trifluoroacetic acid or hydrogen chloride in an organic solvent such as ethyl acetate or dioxane. The condensation reaction between compound (XI) and compound (XII) is an amide bond formation reaction, and compound (II) and compound (III)
The condensation reaction can be performed by the same method.

The conversion reaction from the compound (XIII) to the compound (XIV) can be usually carried out in two steps of deprotection and condensation or substitution reaction. When X ′ is a protected amino group, deprotection is conducted under the same conditions as in the conversion of compound (X) to compound (XI), and then the corresponding carboxylic acid is condensed with compound (II) and compound (III). The condensation reaction can be carried out under the same conditions as described above, or the substitution reaction can be carried out under the same conditions as those used for the substitution reaction of the corresponding halide with the compound (IV) and the compound (V). When X ′ is a protected carboxyl group, it can be removed by a method known per se. For example, methyl or ethyl ester can be removed by reacting it with a base such as sodium hydroxide, potassium hydroxide or lithium hydroxide in an organic solvent such as methanol, ethanol, tetrahydrofuran or dioxane. Further, a compound having a benzyl ester group can be obtained by hydrogenating in the presence of a noble metal catalyst such as palladium or platinum, to obtain a carboxylic acid derivative, and a compound having a tert-butyl ester group can be trifluoroacetic acid, The carboxylic acid derivative can be obtained by treatment with an acid such as hydrogen chloride. The obtained carboxylic acid can be condensed by the method used in the condensation reaction of the corresponding amine compound or hydroxy compound with compound (II) and compound (III) to obtain compound (XIV).

In the above-mentioned process for producing compound (I) and intermediates, the compounds used in the reaction are
For example, inorganic acid salts such as hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc., such as acetate, tartrate, citrate, fumarate, maleate, toluenesulfonate, methanesulfone. It may be used in the form of an organic acid salt such as an acid salt, a metal salt such as a sodium salt, a potassium salt, and an aluminum salt, such as a salt with a base such as a triethylamine salt, a guanidine salt, an ammonium salt, a hydrazine salt, and a quinine salt. Good. When compound (I) is obtained as a free form by the above 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. can do. The compound of formula (I), a salt thereof and a hydrate thereof (in the following description, not only the salt but also the hydrate thereof may be simply referred to as compound (I)) have low toxicity. It is a safe compound that acts on the platelet fibrinogen receptor (glycoprotein (G
Fibrinogen, fibronectin, and von wille for P) IIb / IIIa)
brand factor), and their binding to corresponding receptors on the surface of various types of cells, and of other adhesive proteins such as vitronectin, collagen and laminin.

In producing the microcapsules of the present invention, the amount of the amorphous water-soluble compound (I) used varies depending on the kind of the compound (I), desired pharmacological effect and duration of effect. The concentration of the high molecular weight polymer in the organic solvent solution is about 0.001% to 90% (W / W), more preferably about 0.01% to 80% (W / W), and particularly preferably about 0. It is selected from 0.01% to 70% (W / W). The amorphous water-soluble compound (I) is used in the form of fine particles. The average particle size of the fine particles is generally 30 μm or less,
Usually about 1 nm to about 10 μm is used, preferably 5
It is less than or equal to μm, more preferably about 1 nm to about 1 μm.

The high molecular weight polymer used in the present invention is a biocompatible high molecular weight polymer which is hardly soluble or insoluble in water. Examples thereof include biodegradable polymer, and specific examples thereof include polyfatty acid ester (eg, polylactic acid, polyglycolic acid, polycitric acid, polymalic acid, polylactic acid caprolactone, etc.), poly-α- Cyanoacrylic acid ester, poly-β-hydroxybutyric acid, polyalkylene oxalate (eg, polytrimethylene oxalate, polytetramethylene oxalate, etc.), polyorthoester, polyorthocarbonate, or other polycarbonate (eg, polyethylene carbonate) , Polyethylene propylene carbonate, etc.), polyamino acids (eg, poly-γ-benzyl-L-glutamic acid, poly-
L-alanine, poly-γ-methyl-L-glutamic acid and the like), hyaluronic acid ester and the like. Furthermore, as other high molecular weight polymer having biocompatibility,
Polystyrene, polymethacrylic acid, copolymers of acrylic acid and methacrylic acid, polyamino acids, dextrans tearate, ethyl cellulose, acetyl cellulose, nitrocellulose, maleic anhydride copolymers, ethylene vinyl acetate copolymers, Examples include polyvinyl acetate and polyacrylamide. These high molecular polymers may be one kind, a copolymer of two or more kinds, or a simple mixture, or a salt thereof.

Among these high molecular weight polymers, biodegradable high molecular weight polymers are preferable especially when used as an injection. The biodegradability of the biodegradable polymer is, for example, lactic acid / glycolic acid copolymer (polymer) (PLG).
In the case of A), it is defined as the ratio (w / w%) of the water-soluble low molecular weight fragment decomposed from PLGA to PLGA, and generally 10% or more, preferably subcutaneously or subcutaneously for 3 months after subcutaneous or intramuscular administration. It is 80% or more in one year after intramuscular administration. The biodegradable polymer is preferably polyester. Preferred specific examples of the biodegradable high molecular weight polymer include, for example, a hydroxycarboxylic acid polymer or copolymer, or a mixture thereof. The hydroxycarboxylic acid is not particularly limited,

Embedded image [In the formula, R represents hydrogen or an alkyl group. ] A hydroxycarboxylic acid represented by the following is mentioned as a preferred specific example. In the above formula, the alkyl group represented by R is, for example, a linear or branched alkyl group having 1 to 8 carbon atoms (eg, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl). , Hexyl,
Heptyl, octyl, etc.) are preferred. Among these, a linear or branched alkyl group having 1 to 3 carbon atoms is particularly preferred.

Specific preferred examples of the hydroxycarboxylic acid include glycolic acid, lactic acid, hydroxybutyric acid (eg 2-hydroxybutyric acid), 2'-hydroxyvaleric acid, 2-hydroxy-3-methylbutyric acid and 2-hydroxy. Examples thereof include caproic acid, 2-hydroxyisocaproic acid and 2-hydroxycaprylic acid. Of these, glycolic acid, lactic acid, 2-hydroxybutyric acid, 2-hydroxy-3-methylbutyric acid, and 2-hydroxycaproic acid are particularly preferable. Furthermore, glycolic acid, lactic acid, and 2-hydroxybutyric acid are particularly preferable. Of these hydroxycarboxylic acids, any of those having D-form, L-form and D, L-form may be used, but D, L-form is preferable. The form of copolymerization of the copolymer may be random, block or graft. It is preferable that these glycolic acid copolymers are relatively rapidly decomposed in vivo and have a release period of 1 month or less when used alone. In particular, a lactic acid / glycolic acid copolymer or a homopolymer (hereinafter, also referred to as a homopolymer of either one of the acids may be simply referred to as a copolymer) or a hydroxybutyric acid / glycolic acid copolymer. Is preferred. The high molecular weight polymer used in the present invention can be synthesized by a general synthesis method (see, for example, JP-A-6-63).
No. 1-28521 reference), it can synthesize | combine without a problem.

The weight average molecular weight of these high molecular weight polymers used in the present invention is preferably about 2000 to about 800,000, more preferably about 5000 to about 2.
It is selected from the range of 00000. When a lactic acid / glycolic acid copolymer (polymer) is used as the above-mentioned polymer, the molar ratio of lactic acid / glycolic acid is preferably about 10
0/0 to about 25/75, more preferably about 100 /
0 to about 50/50. The weight average molecular weight of the lactic acid / glycolic acid copolymer is preferably about 5,000 to about 30,000, more preferably about 5,000 to about 2,000.
0. Hydroxybutyric acid as the above polymer
When a glycolic acid copolymer (polymer) (eg, 2-hydroxybutyric acid / glycolic acid copolymer) is used, the hydroxybutyric acid / glycolic acid molar ratio is preferably about 100 /
0 to about 25/75, more preferably about 100/0 to about 50/50. In particular, 2-hydroxybutyric acid /
The molar ratio of glycolic acid is preferably about 60/40 to about 30/70. The weight average molecular weight of the hydroxybutyric acid / glycolic acid copolymer is preferably about 5,000 to about 25,000, more preferably about 5,000 to about 200.
00. When a butyric acid / glycolic acid copolymer is used as the polymer, the butyric acid / glycolic acid molar ratio is preferably about 100/0 to about 25/75.
When a mixture of polylactic acid (A) and glycolic acid / 2-hydroxybutyric acid copolymer (B) is used as the above-mentioned polymer, the mixing ratio represented by (A) / (B) is about 10/90. To about 90/10 (weight ratio). It is preferably in the range of about 25/75 to about 75/25 (weight ratio). The weight average molecular weight of polylactic acid is about 50.
Those from 00 to about 30,000 are preferred. About 60 more
Those from 00 to about 20,000 are particularly preferred.

In the present specification, the molecular weight means a polystyrene-equivalent molecular weight measured by gel permeation chromatography (GPC) using polystyrene as a reference substance. The measurement is GPC column KF804L × 2
(Manufactured by Showa Denko), RI monitor L-3300 (manufactured by Hitachi, Ltd.) were used, and chloroform was used as a mobile phase. More specifically, the weight average molecular weight of the present application is such that the weight average molecular weight is 120,000, 52,000, 22,000, 9,
200, 5,050, 2,950, 1,055, 580,
162 kinds of polystyrene as reference substances, GP
The polystyrene-equivalent molecular weight measured by C. The polydispersity of the high molecular weight polymer is defined as weight average molecular weight / number average molecular weight, and is generally 1 to 3.5, preferably 1.5.
To 2.5. The amount of these high molecular weight polymers used depends on the strength of the pharmacological activity of the physiologically active substance and the release rate and period of the substance. Ratio), preferably about 1 to about 1000 times (weight ratio), is used as the microcapsule base. The concentration of the high molecular weight polymer in the oil phase is about 0.5 to about 90% (W /
W), more preferably about 2 to about 60% (W / W)
Chosen from.

Further, for the purpose of suppressing the initial release of the water-soluble drug from the microcapsules, it is also effective to add a basic substance or fats and oils to the organic solvent solution of the polymer. As the basic substance, for example, L-arginine,
Examples thereof include basic amino acids such as N-methylglucamine and L-lysine. Of these, L-arginine or N-
Methylglucamine is preferred. As fats and oils, for example,
Vitamin E, medium chain fatty acid triglycerides (Miglyol (Dynamite Nobel), etc.), cholesterol, phospholipids and the like can be mentioned. The concentration of the basic substance in the organic solvent solution of the high molecular weight polymer is about 0.01% to about 20% (W / W), preferably about 0.1% to about 5%.
(W / W), more preferably about 0.1% to about 3%
(W / W). The concentration of oils and fats in the organic solvent solution of the high molecular weight polymer is about 0.01% to about 30% (W /
W), preferably about 0.1% to about 20% (W /
W), more preferably about 0.2% to about 10% (W /
W).

In the present invention, it is preferable that the aqueous phase contains an osmotic pressure adjusting agent. As the osmotic pressure adjusting agent,
When used as an aqueous solution, any substance may be used as long as it exhibits an osmotic pressure. Specific examples of the osmotic pressure adjusting agent include, for example, water-soluble polyhydric alcohols, water-soluble monohydric alcohols, water-soluble inorganic substances (eg, inorganic salts), water-soluble monosaccharides, disaccharides, oligosaccharides. And polysaccharides or their derivatives, water-soluble organic acids or salts thereof, water-soluble amino acids, water-soluble peptides, proteins or their derivatives. Of these, water-soluble polyhydric alcohols, water-soluble inorganic substances, water-soluble monosaccharides, disaccharides,
Oligosaccharides and polysaccharides or their derivatives, water-soluble organic acids or salts thereof are preferred. Furthermore, salts, water-soluble polyhydric alcohols and water-soluble inorganic substances are particularly preferable.

Examples of the above water-soluble inorganic salts include alkali metal halides such as potassium chloride, sodium chloride, potassium bromide, sodium bromide, potassium iodide, sodium iodide, etc., halogenated calcium chloride, magnesium chloride, etc. Alkaline earth metal, alkali metal sulfate such as sodium sulfate, potassium sulfate, magnesium sulfate,
Alkaline earth metal sulfates such as calcium sulfate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, alkali metal phosphates such as sodium phosphate, etc. Is mentioned. Of these, sodium chloride is preferred.

The above water-soluble polyhydric alcohols include
Examples thereof include dihydric alcohols such as glycerin, pentahydric alcohols such as arabitol, xylitol and adonitol, and hexahydric alcohols such as mannitol and sorbitol. Of these, hexavalent alcohols are preferable. Examples of the water-soluble monohydric alcohols include methanol, ethanol and isopropyl alcohol. Of these, ethanol is preferred.
Examples of the water-soluble monosaccharides include pentacarbon sugars such as arabinose, xylose, ribose and 2-deoxyribose, and hexacarbon sugars such as glucose, fructose, galactose, mannose, sorbose, rhamnose and fucose. Of these, hexose sugars are preferred. Examples of the water-soluble disaccharides include maltose, cellobiose, α-trehalose, lactose, sucrose and the like. Of these, lactose and sucrose are preferred.

Examples of the water-soluble oligosaccharides include maltotriose, trisaccharides of raffinose sugar, tetrasaccharides such as stachyose, and the like. Of these, trisaccharides are preferred. Examples of the water-soluble polysaccharides include glucans such as cellulose, starch and glycogen, galacturonans such as pectic acid, mannuronans such as alginic acid, fructans such as inulin and levan, and N-acetylglycosamine heavy compounds such as chitin. Examples thereof include coalesced products, xylans such as rice straw xylan, and diheteroglucans such as mannan, glucomannan, galactomannan, hyaluronic acid, chondroitin sulfate, and heparin. Of these, glucans and diheteroglucans are preferable. Examples of the water-soluble monosaccharide, disaccharide, oligosaccharide and polysaccharide derivatives include glucosamine, galactosamine, glucuronic acid and galacturonic acid.

Examples of the water-soluble organic acid or salt thereof include:
Examples thereof include citric acid, tartaric acid, malic acid, alkali metal salts thereof (eg, sodium salt, potassium salt, etc.). Examples of the water-soluble amino acid include neutral amino acids such as glycine, alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, serine, threonine, proline, hydroxyproline, cysteine, methionine, aspartic acid, and glutamic acid. Examples include basic amino acids such as amino acids, lysine, arginine, and histidine. The acid of these water-soluble amino acids (eg hydrochloric acid, sulfuric acid, phosphoric acid, etc.)
Alternatively, a salt with an alkali (eg, an alkali metal such as sodium or potassium) may be used. Examples of the water-soluble peptide, protein or derivative thereof include casein, globulin, prolamin, albumin, gelatin, protamine, histone and the like.

The osmotic pressure adjusting agent may be used alone or in combination of one or more kinds. The concentration of these osmotic pressure adjusting agents in the outer aqueous phase is about 0.001% to about 60% (W / W) when the osmotic pressure adjusting agent is a nonionic substance, and preferably about 0.01%. To about 40% (W / W), more preferably about 0.05 to about 30% (W / W).
When the osmotic pressure adjusting agent is an ionic substance, a concentration obtained by dividing the above concentration by the total ionic value is used. The concentration of the osmotic pressure adjusting agent need not be lower than the solubility, and may be partially dispersed.

The microcapsules of the present invention can be produced, for example, by the s / o / w type in-water drying method shown below. First, the amorphous water-soluble compound (I) is dispersed in a water-insoluble organic solvent solution of a polymer, and the resulting dispersion is mixed well to obtain an s / o type emulsion. In the emulsion, the compound (I) is substantially uniformly dispersed in the polymer solution. The water-soluble compound (I) can be used as it is if it is an amorphous type itself, or can be used after being made amorphous even if it is a crystalline type. Preferably, the amorphous water-soluble compound (I) is obtained from an aqueous solution of the water-soluble compound (I), preferably a dilute aqueous solution, by a rapid drying method such as a freeze drying method and a spray drying method. As mentioned above, the amorphous water-soluble compound (I) is preferably used in the form of fine particles, and the average particle size of the compound (I) is generally about 1 nm to about 3
It is 0 μm, preferably about 1 nm to about 5 μm. When the compound (I) is available in the form of fine particles, it can be used as it is, but when it is not used, it is used after being made fine particles by a conventional method (eg, jet mill method, atomizer method, ball mill method, etc.). The water-insoluble solvent may be any one as long as it dissolves the polymer and is insoluble in water. Specific examples of the water-insoluble solvent include:
For example, halogenated hydrocarbons (eg, dichloromethane, chloroform, dichlorohexane, chloroethane, dichloroethane, trichloroethane, carbon tetrachloride etc.), esters (eg ethyl acetate etc.), ethers (eg ethyl ether etc.), aromatic Hydrocarbons (eg, benzene, toluene etc.), hydrocarbons (eg, n-pentane, n-hexane etc.) and the like can be mentioned. A known dispersion method is used for the emulsification operation of the s / o type emulsion. Examples of the dispersion method include an intermittent shaking method, a method using a mixer such as a propeller-type stirrer or a turbine-type stirrer, a colloid mill method, a homogenizer method, and an ultrasonic irradiation method. In this case, it is also effective to use a water-soluble solvent together with a water-insoluble solvent if desired. The water-soluble solvent may be any one as long as it is water-soluble and can be mixed with the water-insoluble solvent. Specific examples of the water-soluble solvent include:
Examples thereof include alcohols (eg, methanol, ethanol, propyl alcohol, isopropyl alcohol, etc.), acetone, acetonitrile and the like. In the s / o type emulsion, the compound (I) is preferably finely atomized and dispersed, and the particle size of the fine particles is usually about 1 nm to about 30 μm, preferably about 1 nm to about 5 μm, and most preferably about 1 nm. It is 1 nm to about 1 μm.

Then, the s / o type emulsion thus produced is subjected to an underwater drying method in an aqueous phase. Preferably, the aqueous phase contains the above-mentioned concentration of the osmotic pressure adjusting agent. That is, after adding the oil phase to the second phase aqueous phase further containing an osmotic pressure adjusting agent to form an s / o / w type emulsion,
The solvent in the oil phase is removed to produce microcapsules.

An emulsifier may be added to the outer aqueous phase in the s / o / w type underwater drying method. For example, generally stable o
Any material may be used as long as it forms a / w emulsion.
More specifically, for example, anionic surfactants (eg, sodium oleate, sodium stearate, sodium lauryl sulfate, etc.), nonionic surfactants (eg,
Polyoxyethylene sorbitan fatty acid ester [eg, Tween 60, Tween 80 (Atlas Powder Co.), etc.], polyoxyethylene castor oil derivative [eg,
HCO-60, HCO-50 (Nikko Chemicals), etc.]
Alternatively, polyvinylpyrrolidone, polyvinyl alcohol, carboxymethyl cellulose, lecithin, gelatin, etc. may be mentioned, and one of these may be used, or some of them may be used in combination. The concentration at the time of use can be appropriately selected from the range of about 0.01% to about 20% (W / W), and more preferably about 0.05% to about 10%.
Used in the range of (W / W).

For removal of the solvent in the oil phase, a commonly used method is adopted. As the method, a propeller type stirrer,
Alternatively, the pressure is gradually reduced while stirring with a magnetic stirrer or the like, or it is removed while adjusting the degree of vacuum using a rotary evaporator or the like. In this case, when the solidification of the high molecular weight polymer progresses to a certain extent and the loss due to the release of the compound (I) from the inner layer is reduced, the s / o / w type emulsion is used to complete the removal of the solvent. The time required can be shortened by gradually heating. Also,
When thickening and solidifying by methods other than temperature, simply s
The / o / w type emulsion may be removed by leaving it under stirring, heating, blowing nitrogen gas or the like. This process of removing the solvent is an important process that greatly affects the surface structure of the microcapsules that controls the release of compound (I). For example, by increasing the removal rate, many pores are formed on the surface and larger pores are formed, and the release rate of compound (I) is increased.

The microcapsules thus obtained are separated by centrifugation or filtration, and then the free water-soluble compound (I) attached to the surface of the microcapsules,
The compound (I) -retaining substance and the like are repeatedly washed with distilled water several times, and if necessary, heated to completely remove water in the microcapsules and solvent in the microcapsules under reduced pressure. To do. The microcapsules obtained above are lightly crushed if necessary and then sieved to remove microcapsules that are too large. The particle size of the microcapsules, when used as a suspending agent depending on the degree of sustained release, may be in a range that satisfies the dispersibility and needle penetration, for example, an average diameter of about 0.5 to 400 μm. And the range is more preferably about 2 to 200 μm.

The microcapsules produced by the method of the present invention can be administered orally and parenterally.
For example, it can be easily administered as an intramuscular, subcutaneous, blood vessel, organ, or a lesion such as a joint cavity or a tumor as an injection or implant. Further, it can be molded into various preparations for administration, and can be used as a raw material for producing such preparations. The above-mentioned preparations include injections and oral preparations (eg, powders, granules, capsules,
Tablets), nasal preparations, suppositories (eg, rectal suppositories, vaginal suppositories)
And the like. These preparations can be manufactured according to a conventional method. For example, in order to use the microcapsules of the present invention as an injection, the microcapsules of the present invention are used as a dispersant (eg, Tween 80, HCO 60 (manufactured by Nikko Chemicals), carboxymethyl cellulose, sodium alginate, etc.), a preservative. (Eg methylparaben,
Propylparaben, bezyl alcohol, chlorobutanol, etc.), an isotonic agent (eg sodium chloride, glycerin, sorbitol, glucose, etc.) and other aqueous suspensions, or olive oil, sesame oil, peanut oil, cottonseed oil, corn oil, etc. It is dispersed in vegetable oil, propylene glycol or the like to be molded into an oily suspension to give a sustained release injection.

Further, the sustained-release injection of microcapsules described above was redispersed by adding an excipient (eg, mannitol, sorbitol, lactose, glucose, etc.) as a suspension, in addition to the above composition. Thereafter, it is freeze-dried or spray-dried to solidify, and distilled water for injection or a suitable dispersant is added at the time of use to obtain a more stable sustained-release injection.

The microcapsules of the present invention can be formed into tablets, for example, by a generally known production method. For example, excipients (eg, lactose, crystalline cellulose, sucrose,
Starch such as corn starch), disintegrant (eg starch such as corn starch, croscarmellose sodium, sodium carboxymethyl starch, calcium carbonate, etc.), binder (eg crystalline cellulose, gum arabic, dextrin, carboxy) Methylcellulose, polyvinylpyrrolidone, hydroxypropylcellulose, etc. or lubricants (eg talc, magnesium stearate, polyethylene glycol 600)
0, etc.) is added and compression molding is performed.

The microcapsules of the present invention can be formed into a solid, semi-solid, or liquid form by, for example, nasal administration formulation, and any of them can be produced by a commonly used production method. For example, as the solid form, the microcapsules may be used as they are, or an excipient (eg, glucose, mannitol, starch, microcrystalline cellulose, etc.), a thickener (eg, natural gums, cellulose derivatives, acrylic acid). Polymers, etc.) are added and mixed to obtain a powdery composition.
The liquid is almost the same as the injection, and is an oily or aqueous suspension. If it is semi-solid,
Aqueous or oily gels or ointments are preferred.
In addition, all of these are pH regulators (eg, carbonic acid, phosphoric acid, citric acid, hydrochloric acid, sodium hydroxide, etc.), preservatives (eg, paraoxybenzoic acid esters, chlorobutanol, benzalkonium chloride, etc.), etc. May be added.

In order to use the microcapsules of the present invention as suppositories, oily or aqueous solid, semi-solid or liquid products are prepared by a method known per se using an oily base or an aqueous base, respectively. sell. The oily base used in the composition may be one that does not dissolve the microcapsules,
For example, glycerides of higher fatty acids [eg, cacao butter, Waitepsols (Dynamite Nobel), etc.], medium-chain triglycerides [eg, Miglyols (Dynamite Nobel), etc.], or vegetable oils (eg, sesame oil, soybean oil, cottonseed oil, etc.) ) And the like. Examples of the aqueous base include polyethylene glycols and propylene glycol, and examples of the aqueous gel base include natural gums, cellulose derivatives, vinyl polymers and acrylic acid polymers.

Since the microcapsules of the present invention release a constant drug amount over a long period of time, they are expected to be safe and highly effective sustained-release preparations which have low toxicity and constant drug efficacy. For example, even when bleeding tendency is a concern as a side effect of antithrombotic action, the use of the microcapsule of the present invention makes it possible to maintain a non-toxic and effective concentration in a region where side effects are suppressed for a long period of time. Therefore, in the microcapsule of the present invention, the compound (I) is a glycoprotein IIb as described above.
/ IIIa, such as fibrinogen, inhibits their adhesion proteins and other adhesion proteins from their corresponding receptors on the surface of various types of cells and inhibits cell-cell and cell-matrix interactions. Angina of mammals including human (e.g., mouse, rat, guinea pig, dog, rabbit, human, etc.) because it affects the action, and particularly the compound inhibits the formation of thrombus.
Unstable angina, acute myocardial infarction, Kawasaki disease, acute and chronic heart failure, transient cerebral ischemic attack (TIA), stroke, cerebral ischemic injury in acute cerebral thrombosis, dissecting aneurysm, subarachnoid Cerebral vasospasm after bleeding, acute or chronic renal disease (eg acute or chronic renal disease due to over-aggregation such as snake venom and immune diseases), chronic and acute glomerulonephritis, diabetic nephritis and neuropathy, nephrotic syndrome, liver Disease, pulmonary embolism, bronchial asthma, pulmonary edema, adult respiratory distress syndrome (ARDS), arteriosclerosis obliterans, peripheral arterial occlusion, deep vein thrombosis, oscillatory disease, peripheral arterial occlusion with diabetes, thrombotic thrombocytopenic purpura Disease (TTP), disseminated intravascular coagulation (DIC), sepsis, surgical or infectious shock, post- and postpartum trauma,
Placental premature detachment, incompatible transfusion, systemic lupus erythematosus,
It can be used for treatment or prevention of Raynaud's disease, inflammation, arteriosclerosis, hemolytic uremic syndrome, targeted peripheral artery necrosis, humiliation, and hemorrhoidal disease. Furthermore, the microcapsules of the present invention are used for cardiac bypass surgery, cardiopulmonary bypass, atrial fibrillation,
It can be used for surgery associated with hip fracture, valve replacement, prevention of thrombosis due to artificial blood vessels and organs, prevention of thrombocytopenia by artificial dialysis, and secondary prevention of myocardial infarction. Here, prevention of thrombocytopenia by artificial dialysis also means prevention of coagulation and residual blood in a hemodialysis extracorporeal circuit. Furthermore, the microcapsule of the present invention is used for coronary thrombolytic therapy (for example, enhancement of action of thrombolytic agents such as tissue plasminogen activator (TPA) and prevention of reocclusion), PTCA (percutaneous intracoronary angioplasty), stent placement, and Can be used to prevent reocclusion and restenosis of coronary arteries after atherectomy, prevent reocclusion and restenosis after coronary artery bypass surgery, and prevent ischemic complications (eg myocardial infarction, death) during PTCA or coronary thrombolytic therapy Besides, it can be used as an antitumor agent by inhibiting cancer metastasis. In particular, the microcapsule of the present invention is useful for the prevention / treatment of thrombosis, angina, unstable angina and PTCA or ischemic complication after coronary thrombolytic therapy, and coronary reocclusion and restenosis.

The dose of the sustained-release preparation of the present invention includes the type and content of the main compound (I), the dosage form, the duration of drug release, the animal to be administered (eg, mouse, rat, horse,
It depends on the purpose of administration, such as warm-blooded mammals such as cows and humans), and the effective amount of the main drug is sufficient. For example, when orally administered to patients with unstable angina, PTCA or patients undergoing coronary thrombolytic therapy for ischemic complications or reocclusion or restenosis of coronary arteries, the dose per adult (body weight 50 kg) The suitable amount of the microcapsules is in the range of about 1 mg to 10 g, preferably about 10 mg to 2 g (usually about 1 to 50 per dose as the main compound (I)).
It can be appropriately selected from 0 mg, preferably in the range of about 10 to 200 mg). In addition, suspension when administered parenterally to patients with unstable angina, ischemic complications after PTCA or coronary thrombolytic therapy, or patients with reocclusion or restenosis of coronary arteries, for example, when administered as the above injection The volume of the solution is in the range of about 0.1 to 5 ml, preferably about 0.5 to 3 ml (usually an adult (body weight 50 kg)).
Can be appropriately selected from the range of about 0.05 to 50 mg, preferably about 1 to 20 mg as the compound (I) which is the main drug). In this manner, the effective amount of the water-soluble compound (I) and the biocompatible high molecular weight polymer, which are larger than the usual single dose, can be used to continuously release the compound (I) over a long period of time. A pharmaceutical composition prepared as possible microcapsules is obtained.

The microcapsules of the present invention have the following features, for example. (1) By using an amorphous water-soluble drug, the drug can be efficiently incorporated into the microcapsule as compared with the microcapsule using the crystalline drug. (2) The initial release amount of the drug after administration of microcapsules can be reduced, and side effects such as bleeding can be suppressed. (3) By using a microcapsule having a high drug content, the total dose of the preparation can be reduced, so that, for example, pain and local irritation at the subcutaneous administration site can be alleviated.

[0075]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described more specifically with reference to Experimental Examples, Examples and Reference Examples, but the present invention is not limited to these exemplifications. The percentages used in the examples are weight / weight percentages unless otherwise stated.

Experimental Example Crystalline compound A was subjected to the same method as in Example 1 below to prepare crystalline compound A-containing microcapsules, which were compared with the amorphous compound A-containing microcapsules obtained in Example 1. . The drug encapsulation rate and the in vitro release property of the obtained microcapsules were measured, and the results shown in [Table 1] were obtained.

[Table 1] Drugs Drug encapsulation rate Initial release amount per day Amorphous 78% 9% Crystal 67% 46% From the above results, more amorphous drug is encapsulated in microcapsules than crystalline drug. Moreover, it can be seen that the initial release is suppressed.

【Example】

Example 1 Fine powder of amorphous compound A (450 mg) obtained by freeze-drying
A lactic acid / glycolic acid copolymer (lactic acid / glycolic acid = 75/25, polystyrene-equivalent weight average molecular weight 10200) in which L-arginine (90 mg) was dissolved in advance was dispersed in 4.0 ml of a solution of 4.05 g of methylene chloride, and Polytron [Kinematica Co., Ltd. (Switzerland)]. The mixture was then emulsified using a homogenizer in 800 ml of a 0.1% aqueous polyvinyl alcohol solution containing 2.7% sodium chloride cooled to 15 ° C. The obtained s / o / w type emulsion is
The mixture was slowly stirred with an ordinary propeller stirrer for 3 hours, waited for the microcapsules to solidify as the methylene chloride volatilized, was collected by centrifugation, and was washed with purified water. The collected microcapsules were obtained as a powder by freeze-drying overnight. The drug encapsulation rate and the in vitro release rate of the obtained microcapsules were measured. The drug trap rate was 78%, and the initial daily release amount was 9%.
%Met.

Example 2 Method s / o / w: Compound A (60 mg) as a fine powder obtained by freeze-drying
Is a lactic acid / glycolic acid copolymer (lactic acid / glycolic acid = 75/25, polystyrene-converted weight average molecular weight 102
(00) was dispersed in a solution of 1.94 g of methylene chloride in 2 ml and atomized with a polytron. Then cooled to 15 ° C. 2.7
The mixture was emulsified using a homogenizer in 800 ml of a 0.1% aqueous polyvinyl alcohol solution containing 10% sodium chloride. The obtained s / o / w emulsion was subjected to the same operations as in Example 1 to prepare compound A-containing microcapsules. w / o / w method: Compound A (60 mg) was dissolved in 1 ml of a 1% acetic acid aqueous solution to form an inner aqueous phase, and the lactic acid / glycolic acid copolymer (lactic acid / glycolic acid = 75/25, polystyrene-equivalent average molecular weight) 10200) 1.94 g of methylene chloride 2 ml
It was mixed with the solution and atomized with a Polytron to give a w / o emulsion. It was then emulsified using a homogenizer in 800 ml of a 0.1% aqueous polyvinyl alcohol solution containing 2.7% sodium chloride cooled to 15 ° C. The obtained w / o / w method emulsion was subjected to the same operations as in Example 1 to prepare compound A-containing microcapsules. The in vitro release properties of the microcapsules obtained in the above s / o / w type and w / o / w type were measured, and the initial daily release amounts were 16% and 33%, respectively.
The microcapsules obtained by the s / o / w method of the present invention were able to suppress the initial release.

Example 3 Compound B (450 mg), which is a fine powder obtained by freeze-drying, was added to L in advance.
A dispersion of arginine (90 mg) in lactic acid / glycolic acid copolymer (lactic acid / glycolic acid = 50/50, polystyrene-equivalent weight average molecular weight 9200) (3.96 g) in methylene chloride (4 ml) was dispersed in a polytron. It was then emulsified using a homogenizer in 800 ml of 0.2% aqueous polyvinyl alcohol solution containing 2.7% sodium chloride cooled to 15 ° C. Obtained s / o
The / w emulsion was subjected to the same operations as in Example 1 to prepare compound B-containing microcapsules.

Example 4 Fine powder of compound C (150 mg) obtained by spray drying was added to L in advance.
-Lactic acid / glycolic acid copolymer in which lysine (90 mg) was dissolved (lactic acid / glycolic acid = 50/50, polystyrene equivalent weight average molecular weight 8000) 4.26 g of methylene chloride 4.
It was dispersed in a 5 ml solution and atomized with a Polytron. Then
The mixture was emulsified using a homogenizer in 800 ml of a 0.2% aqueous polyvinyl alcohol solution containing 0.9% sodium chloride cooled to 15 ° C. The resulting s / o / w emulsion was slowly stirred with an ordinary propeller stirrer for 3 hours, collected by centrifugal separation after waiting for the microcapsules to solidify with the evaporation of methylene chloride, and then purified water. Washed with water. The collected microcapsules were obtained as a powder by freeze-drying with mannitol overnight.

EXAMPLE 5 Hydroxybutyric acid / glycolic acid copolymer (hydroxybutyric acid / glycolic acid = 75/25, polystyrene-converted weight average molecular weight 12000) of 4.20 g of fine powder of compound D (300 mg) obtained by freeze-drying Was dispersed in a solution of methylene chloride (5 ml) and atomized with a polytron. The mixture was then emulsified using a homogenizer in 1000 ml of a 0.2% aqueous polyvinyl alcohol solution containing 1.8% sodium chloride cooled to 15 ° C. The obtained s / o / w type emulsion was subjected to the same operations as in Example 4 to prepare compound D-containing microcapsules.

Example 6 Compound E (200 mg) as a fine powder obtained by freeze-drying was preliminarily diluted with N
-Methylglucamine (100 mg) dissolved in lactic acid / glycolic acid copolymer (lactic acid / glycolic acid = 90/10, polystyrene equivalent weight average molecular weight 8400) Dispersed in 3.70 g of methylene chloride 4 ml solution, and atomized with Polytron. did. Then, 800 ml of a 0.1% polyvinyl alcohol aqueous solution containing 10% mannitol cooled to 15 ° C.
And emulsified using a homogenizer. Got s
The / o / w type emulsion was subjected to the same operations as in Example 4 to prepare compound E-containing microcapsules.

Reference Example 1 (S) -3- (3-t-Butoxycarbonylaminopropyl) -2-oxopiperazine-1
-Acetic acid t-butyl ester oxalate (2,2-dimethoxyethyl) aminoacetic acid t-butyl ester (6.0 g 27.7 mmol), N-Z-Orn (Boc) -O
H (10.0 g 27.7 mmol) was dissolved in 54.6 cc of acetone and stirred at 15 ° C. 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (5.6 g 29.9).
2 mmol) was added. After stirring at room temperature for 1 hour, the reaction system was concentrated under reduced pressure, the residue was dissolved in ethyl acetate and washed with 5% aqueous potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate. The organic layer was concentrated under reduced pressure to obtain a pale yellow oily substance. This oil and p-toluenesulfonic acid 1.0 hydrate (1.04 g 5.46 mmol) were dissolved in 137 cc of toluene, and the mixture was stirred at 70 ° C. for 2 hours. The reaction system was poured into saturated aqueous sodium hydrogen carbonate, and the layers were separated and extracted with ethyl acetate. The organic layer was concentrated under reduced pressure and purified by silica gel column chromatography (hexane / ethyl acetate = 3/2) to obtain 8.3 g of a pale yellow oily substance. This oil (8.3
g (16.5 mmol) was dissolved in 166 cc of ethyl acetate, and 1
1.7 g of 0% Pd-C was added, and the mixture was stirred under a hydrogen atmosphere for 2 hours. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. Dissolve the residue in 16.6cc of methanol and add oxalic acid 2.0 hydrate.
(2.1 g 16.5 mmol) was added, and the mixture was concentrated under reduced pressure. The precipitated crystals were washed with ethyl acetate to give the title compound (5.1 g, 6 g)
6.8%) was obtained as white crystals. Specific rotation: [α] _D −29.3 ° (C = 0.73, H ₂ O) Melting point: 181 ° C. Elemental analysis: C ₁₈ H ₃₃ N ₃ O ₅ · (CO ₂ H) ₂ (461. 51
Calculated as 1): C, 52.05; H, 7.64; N, 9.10 Found: C, 51.98; H, 7.61; N, 9.20

Reference Example 2 (S) -4-benzyloxycarbonylaminoacetyl-3- (3-t-butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid
t-Butyl ester (S) -3- (3-t-butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid t-butyl ester oxalate (1.6 g 3.47 mmol) was dissolved in saturated sodium bicarbonate water. The mixture was separated, extracted with ethyl acetate, and concentrated under reduced pressure.
Residue and NZ-Gly-OH (0.87g 4.16mmol)
Was dissolved in 16.0 cc of acetone, and 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (0.87 g, 4.51 mmol) was added with stirring at 15 ° C. After stirring at room temperature for 1 hour, the reaction system was concentrated under reduced pressure, the residue was dissolved in ethyl acetate and washed with 5% aqueous potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate. The organic layer was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate) to give the title compound (1.95 g, 100%) as a colorless amorphous powder. IR ν max cm ^-1 : 3360,2970,2930,1
713, 1650, 1513, 1448, 1363, 124
6,1158,1045,964,848,744,695 NMR (CDCl ₃ ) δ: 1.43 (9H, s), 1.46 (9H,
s), 1.50-2.20 (4H, m), 3.02-4.28 (10
H, m), 4.52-4.80 (1H, m), 5.01 (1H, dd, J
= 8.8Hz, J = 4.6Hz), 5.13 (2H, s), 5.64-
5.86 (1H, m), 7.37 (5H, s)

Reference Example 3 (S) -4- (4-amidinobenzoyl) aminoacetyl-3- (3-aminopropyl)-
2-oxopiperazine-1-acetic acid trifluoroacetate (S) -4-benzyloxycarbonylaminoacetyl-
3- (3-t-butoxycarbonylaminopropyl) -2
-Oxopiperazine-1-acetic acid t-butyl ester
(1.34 g 2.38 mmol) was dissolved in 13.4 cc of methanol, 0.54 g of 10% Pd-C was added, and the mixture was mixed with hydrogen under an atmosphere of 3
Stir for 0 minutes. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue and baking soda (0.4 g 4.76 mmol) were added to H ₂ O 2
It was dissolved in 6.8 cc and 1,4-dioxane 13.4 cc, and 4-amidinobenzoyl chloride hydrochloride (0.
68 g (3.09 mmol) were added. After stirring for 3 hours, the reaction system was adjusted to PH4 with 1N hydrochloric acid water and concentrated to dryness. Dissolve the residue in 3.75 cc of trifluoroacetic acid and mix at room temperature for 2
Stirred for hours. The reaction system was concentrated under reduced pressure and purified by CHP-20 (Mitsubishi Kasei) column chromatography (H ₂ O),
1.0 g (63.3%) of the title compound was obtained as a colorless amorphous powder. Specific rotation: [α] _D + 35.4 ° (C = 0.75, MeOH) Elemental analysis value: C ₁₉ H ₂₆ N ₆ O ₅ .2CF ₃ CO ₂ H.H ₂ O
Calculated as (664.515): C, 41.57; H, 4.55; N, 12.65 Found: C, 41.86; H, 4.50; N, 12.60

Reference Example 4 (S) -4- (4-amidinobenzoyl) aminoacetyl-3- {3- (4-amidinobenzoyl) amino} propyl-2-oxopiperazine-1-
Acetic acid trifluoroacetate (ie compound B) (S) -4- (4-amidinobenzoylamino) acetyl-
3- (3-aminopropyl) -2-oxopiperazine-1
Acetic acid trifluoroacetic acid salt (0.5 g 0.94 mmol) and sodium bicarbonate (0.32 g 3.76 mmol) in H ₂ O 5.0 cc, 1,4
-Dissolved in 2.5 cc of dioxane and stirred at room temperature with 4-amidinobenzoyl chloride hydrochloride (0.22 g 0.99 mm
ol). After stirring for 2 hours, the reaction system was adjusted to PH4 with 1N HCl water and then concentrated under reduced pressure. The residue is CHP-2
Purification by 0 column chromatography (H ₂ O → 5% CH ₃ CN) gave the title compound 0.34 g (50.7%) as a colorless amorphous powder. Specific rotation: [α] _D + 41.9 ° (C = 0.73, MeOH) Elemental analysis value: C ₂₇ H ₃₂ N ₈ O ₆ · CF ₃ CO ₂ H · 2H ₂ O
Calculated as (714.653): C, 48.74; H, 5.22; N, 15.68 Found: C, 48.52; H, 5.22; N, 15.57

Reference Example 5 (S) -3- (4-t-Butoxycarbonylaminobutyl) -2-oxopiperazine-1-
Acetic acid t-butyl ester oxalate By the same method as in Reference Example 1, NZ-Lys (Boc) -OH
Was synthesized using Specific rotation: [α] _D −29.0 ° (C = 1.02, DMS
O) Melting point: 170-172 ° C. Elemental analysis value: C ₁₉ H ₃₅ N ₃ O _5. (CO ₂ H) ₂ (475.54
0) calculated: C, 53.04; H, 7.84; N, 8.84 Found: C, 52.75; H, 7.65; N, 8.66

Reference Example 6 (S) -4-benzyloxycarbonylaminoacetyl-3- (4-t-butoxycarbonylaminobutyl) -2-oxopiperazine-1-acetic acid t
-Butyl ester (S) -3- (4-t-butoxycarbonylaminobutyl) -2-oxopiperazine- by the same method as in Reference Example 2.
It was synthesized using 1-acetic acid t-butyl ester oxalate. IR ν max cm ^-1 : 3400, 2990, 2945, ¹
713, 1657, 1520, 1458, 1368, 125
3,1166,1070,745,700 NMR (CDCl ₃ ) δ: 1.42 (9H, s), 1.46 (9H,
s), 1.18-2.12 (6H, m), 2.92-4.28 (10
H, m), 4.48-4.84 (1H, m), 5.02 (1H, dd, J
= 8.6Hz, J = 4.8Hz), 5.13 (2H, s), 5.60-
5.88 (1H, m), 7.36 (5H, s)

Reference Example 7 (S) -4- (4-amidinobenzoylamino) acetyl-3- (4-aminobutyl) -2
-Oxopiperazine-1-acetic acid trifluoroacetic acid salt By the method similar to Reference Example 3, (S)-[4-benzyloxycarbonylaminoacetyl-3- (4-t-butoxycarbonylamino-butyl) -2-oxopiperazine -1
-Yl] -acetic acid t-butyl ester. Specific rotation: [α] _D + 46.8 ° (C = 1.01, H ₂ O) Elemental analysis value: C ₂₀ H ₂₈ N ₆ O ₅・ 1.7CF ₃ CO ₂ H ・ 2H
Calculated as ₂ O (662.394): C, 42.43; H, 5.13; N, 12.69 Found: C, 42.53; H, 4.88; N, 12.78.

Reference Example 8 (S) -4- (4-amidinobenzoylamino) acetyl-3- {4- (4-amidinobenzoylamino) butyl} -2-oxopiperazine-1-acetic acid 1 trifluoroacetic acid salt 1 Hydrochloride (S) -4- (4-amidinobenzoylamino) acetyl-3- (4-aminobutyl) was prepared in the same manner as in Reference Example 4.
2-oxopiperazine-1-acetic acid Synthesized using trifluoroacetic acid salt. Specific rotation: [α] _D + 44.3 ° (C = 1.01, H ₂ O) Elemental analysis value: C ₂₈ H ₃₄ N ₈ O ₆ · CF ₃ CO ₂ H · HCl · 3
Calculated as H ₂ O (78.157): C, 46.01; H, 5.41; N, 14.31 Found: C, 46.23; H, 5.22; N, 14.54.

Reference Example 9 (S, S) -4- {2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl} -3- (3-t-butoxycarbonylaminopropyl) -2-oxopiperazine -1-Acetic acid t-butyl ester NZ-Tyr (OMe) -O was prepared in the same manner as in Reference Example 2.
Synthesized using H. IR ν max cm ^-1 : 3360, 2975, 2925, ¹
710,1643,1512,1448,1360,124
5,1152,1033,743,696 NMR (CDCl ₃ ) δ: 1.41 (9H, s), 1.44 (9H,
s), 1.30-2.10 (3H, m), 2.20-2.44 (1H,
m), 2.80-3.84 (10H, m), 3.77 (3H, s), 4.
23 (1H, d, J = 17.2Hz), 4.50-4.85 (1H,
m), 4.93 (1H, dd, J = 6.2Hz, J = 7.0Hz), 5.
09 (2H, dd, J = 12.0Hz, J = 16.4Hz), 5.6
7 (1H, d, J = 8.8Hz), 6.80 (2H, d, J = 8.8H)
z), 7.09 (2H, d, J = 8.8Hz), 7.35 (5H, s)

Reference Example 10 (S, S) -4- {2- (4-amidinobenzoylamino) -3- (4-methoxyphenyl) -propionyl} -3- (3-aminopropyl) -2-
Oxopiperazine-1-acetic acid trifluoroacetic acid salt By the same method as in Reference Example 3, (S, S) -4- {2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl} -3- (3- t-butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid t-
Synthesized using butyl ester. Specific rotation: [α] _D + 78.2 ° (C = 0.62, H ₂ O) Elemental analysis value: C ₂₇ H ₃₄ N ₆ O ₆・ CF ₃ CO ₂ H ・ 3H ₂ O
Calculated as (706.672): C, 49.29; H, 5.85; N, 11.89 Found: C, 49.53; H, 5.68; N, 11.90

Reference Example 11 (S, S) -4- {2- (4-amidinobenzoylamino) -3- (4-methoxyphenyl) -propionyl} -3- {3- (4-amidinobenzoylamino) propyl } -2-Oxopiperazine-1-acetic acid trifluoroacetic acid salt By the same method as in Reference Example 4, (S, S) -4- {2- (4-
Amidinobenzoylamino) -3- (4-methoxyphenyl) propionyl} -3- (3-aminopropyl) -2-oxopiperazine-1-acetic acid was synthesized using trifluoroacetic acid salt. Specific rotation: [α] _D + 52.8 ° (C = 0.76, MeOH) Elemental analysis: C ₃₅ H ₄₀ N ₈ O ₇ · CF ₃ CO ₂ H · 3H ₂ O
Calculated as (852.820): C, 52.11; H, 5.55; N, 13.14 Found: C, 52.27; H, 5.50; N, 13.26

Reference Example 12 (S, S) -4- {2-benzyloxycarbonylamino-3- (4-methoxyphenyl)
-Propionyl} -3- (4-t-butoxycarbonylaminobutyl) -2-oxopiperazine-1-acetic acid t-butyl ester By the same method as in Reference Example 2, (S) -3- (4-tbutoxycarbonylamino) Butyl) -2-oxopiperazine-1
-Acetic acid t-butyl ester oxalate and N-Z-Ty
Synthesized using r (OMe) -OH. IR ν max cm ^-1 : 3345,2975,2930,1
712,1646,1512,1447,1364,124
4,1155,1034,743,696 NMR (CDCl ₃ ) δ: 1.43 (9H, s), 1.44 (9H,
s), 1.00-2.45 (6H, m), 2.80-3.90 (10
H, m), 3.78 (3H, s), 4.23 (1H, d, J = 17.4H
z), 4.70-5.10 (2H, m), 5.10 (2H, d, J = 2.
4Hz), 5.74 (1H, d, J = 8.8Hz), 6.81 (2H,
d, J = 8.6Hz), 7.10 (2H, d, J = 8.6Hz), 7.3
5 (5H, s)

Reference Example 13 (S, S) -4- {2- (4-amidinobenzoylamino) -3- (4-methoxyphenyl) propionyl} -3- (4-aminobutyl) -2-oxopiperazine- 1-Acetic acid trifluoroacetic acid salt By the same method as in Reference Example 3, (S, S)-[4- {2-benzyloxycarbonylamino-3- (4-methoxyphenyl) -propionyl} -3- (4-t -Butoxycarbonylaminobutyl) -2-oxopiperazin-1-yl]-
Synthesized using acetic acid t-butyl ester. Specific rotation: [α] _D + 53.1 ° (C = 0.64, MeOH) Elemental analysis value: C ₂₈ H ₃₆ N ₆ O ₆ · CF ₃ CO ₂ H · 3H ₂ O
Calculated as (720.699): C, 50.00; H, 6.01; N, 11.66 Found: C, 49.87; H, 5.77; N, 11.45

Reference Example 14 (S, S) -4- {2- (4-amidinobenzoylamino) -3- (4-methoxyphenyl) propionyl} -3- {4- (4-amidinobenzoylamino) butyl} 2-oxopiperazine-1-acetic acid hydrochloride By the same method as in Reference Example 4, (S, S) -4- {2- (4-
Amidinobenzoylamino) -3- (4-methoxyphenyl) propionyl} -3- (4-aminobutyl) -2-oxopiperazine-1-acetic acid was synthesized using trifluoroacetic acid salt. Specific rotation: [α] _D + 54.5 ° (C = 0.88, H ₂ O) Elemental analysis value: C ₃₆ H ₄₂ N ₈ O ₇ · HCl · 6H ₂ O (843.
329) as calculated: C, 51.27; H, 6.57; N, 13.29 Found: C, 51.24; H, 6.37; N, 13.26

Reference Example 15 (S) -4-benzyloxycarbonylaminoacetyl-3- {3- (6-t-butoxycarbonylaminohexanoylamino) propyl} -2-
Oxopiperazine-1-acetic acid (S) -4-benzyloxycarbonylaminoacetyl-3- (3-t-butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid t-butyl ester (obtained in Reference Example 2) 0.6 g 1.07 mmol) trifluoroacetic acid 3.
After dissolving in 0 cc and stirring at room temperature for 30 minutes, the reaction system was concentrated under reduced pressure. 6-t-butoxyaminocaproic acid (0.2
6 g 1.12 mmol), 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (0.22 g 1.
14 mmol) and 1-hydroxybenzotriazole
Dissolve (0.15g 1.12mmol) in 2.1cc DMF 1
Stirred for hours. To this solution was added 2.1 cc of DMF solution in which the above residue and triethylamine (0.3 cc 2.14 mmol) were dissolved, and the mixture was stirred at room temperature for 4 hours. The reaction system was diluted with ethyl acetate and washed with 5% aqueous potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate. The organic layer was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate / methanol / acetic acid = 20/10 / 0.6) to give 0.42 g (yield 63.3%) of the title compound as colorless amorphous. Obtained as a crystalline powder. IR ν max cm ^-1 : 3320, 2930,164
3,1533,1448,1203,1173,1046 NMR (CDCl ₃ ) δ: 1.42 (9H, s), 1.20-2.0
9 (10H, m), 2.17 (2H, t, J = 7.3Hz), 2.92
-4.20 (12H, m), 4.80-4.98 (1H, m), 5.1
1 (2H, s), 7.22-7.44 (5H, m)

Reference Example 16 (S) -4- (4-amidinobenzoylamino) acetyl-3- {3- (6-aminohexanoylamino) propyl} -2-oxopiperazine-1
-Acetic acid trifluoroacetate (S) -4-benzyloxycarbonylaminoacetyl-
3- {3- (6-t-Butoxycarbonylaminohexanoylamino) propyl} -2-oxopiperazine-1-acetic acid (0.42 g 0.68 mmol) was dissolved in 8.4 cc of methanol and 10% Pd-CO was added. Add .17g and under hydrogen atmosphere 1
Stirred for hours. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue and sodium bicarbonate a (0.18g 2.14mmol) H ₂ O8.
4 cc, 1,4-dioxane 4.2 cc, 4-amidinobenzoyl chloride hydrochloride (0.20 g) was stirred at room temperature.
0.93 mmol) was added. After stirring for 1 hour, the reaction system was 1N
After adjusting to pH4 with HCl water, the mixture was concentrated to dryness. Dissolve the residue in 4.3 cc of trifluoroacetic acid and add 1 at room temperature.
Stirred for hours. The reaction system is concentrated under reduced pressure, and the residue is CHP-2.
Purification by 0 column chromatography (H ₂ O → 5% CH ₃ CN) gave the title compound (0.26 g, yield 55%) as a colorless amorphous powder. Specific rotation: [α] _D + 42.7 ° (C = 0.99, MeOH) Elemental analysis value: C ₂₅ H ₃₇ N ₇ O ₆ · 1.1CF ₃ CO ₂ H · 2H
Calculated as ₂ O (693.067): C, 47.14; H, 6.12; N, 14.15 Found: C, 47.30; H, 5.82; N, 14.40.

Reference Example 17 (S) -4-Benzyloxycarbonylaminoacetyl-3- {3- (5-t-butoxycarbonylaminopentanoylamino) propyl} -2-
Oxopiperazine-1-acetic acid It was synthesized using 5-t-butoxyaminovaleric acid by the same method as in Reference Example 15. IR ν max cm ^-1 : 3370,2940,1650,1
533, 1455, 1254, 1170, 1050 NMR (CDCl ₃ ) δ: 1.42 (9H, s), 1.28-2.0
8 (8H, m), 2.18 (2H, t, J = 7.0Hz), 3.03 (2
H, t, J = 6.8Hz), 3.10-4.20 (10H, m), 4.
82-5.00 (1H, m), 5.11 (2H, s), 7.22-7.
52 (5H, m)

Reference Example 18 (S) -4- (4-amidinobenzoylamino) acetyl-3- {3- (5-aminopentanoylamino) propyl} -2-oxopiperazine-1
-Acetic acid trifluoroacetic acid salt (S) -4-benzyloxycarbonylaminoacetyl-3- {3- (5-t-butoxycarbonylaminopentanoylamino) propyl}-by the same method as in Reference Example 16.
Synthesized using 2-oxopiperazine-1-acetic acid. Specific rotation: [α] _D + 46.0 ° (C = 1.01, MeOH) Elemental analysis value: C ₂₄ H ₃₅ N ₇ O ₆ · CF ₃ CO ₂ H · 2.5H ₂
Calculated as O (676.646): C, 46.15; H, 6.11; N, 14.49 Found: C, 46.43; H, 6.15; N, 14.20.

Reference Example 19 (S) -4-Benzyloxycarbonylaminoacetyl-3- {3- (4-t-butoxycarbonylaminobutanoylamino) propyl} -2-oxopiperazine-1-acetic acid Reference Example 15 and A similar method was used to synthesize 4-t-butoxyaminobutyric acid. IR ν max cm ^-1 : 3350,2930,1642,1
530, 1452, 1252, 1170, 1050 NMR (CDCl ₃ ) δ: 1.42 (9H, s), 1.30-2.1
0 (4H, m), 1.73 (2H, t, J = 7.2Hz), 2.18 (2
H, t, J = 7.5Hz), 3.04 (2H, t, J = 6.8Hz),
3.10-4.20 (10H, m), 4.83-4.97 (1H,
m), 5.11 (2H, s), 7.22-7.50 (5H, m)

Reference Example 20 (S) -4- (4-amidinobenzoylamino) acetyl-3- {3- (4-aminobutanoylamino) propyl} -2-oxopiperazine-1-
Acetic acid trifluoroacetate (S) -4-benzyloxycarbonylaminoacetyl-3- {3- (4-t-butoxycarbonylaminobutanoylamino) propyl} -2 by the same method as in Reference Example 16.
Synthesized using -oxopiperazine-1-acetic acid. Specific rotation: [α] _D + 47.9 ° (C = 1.00, H ₂ O) Elemental analysis value: C ₂₃ H ₃₃ N ₇ O ₆・ 1.5CE ₃ CO ₂ H ・ 2H
Calculated as ₂ O (710.623): C, 43.95; H, 5.46; N, 13.80 Found: C, 44.23; H, 5.63; N, 13.52

Reference Example 21 (S, S) -4- {2- (4-amidinobenzoylamino) -3- (4-methoxyphenyl) propionyl} -3- (4-guanidinobutyl) -2-
Oxopiperazine-1-acetic acid hydrochloride (S, S) -4- {2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl} -3- (4-t-butoxycarbonyl) obtained in Reference Example 12 Aminobutyl)
2-oxopiperazine-1-acetic acid t-butyl ester (0.6 g 0.86 mmol) was dissolved in 2.0 cc of trifluoroacetic acid. After stirring at room temperature for 1 hour, the reaction system was concentrated under reduced pressure. An aqueous solution (5.6 cc) of the residue and sodium bicarbonate (0.22 g, 2.57 mmol) was added to S-methylisothiourea sulfate (0.4).
8 g 1.71 mmol) and 2N NaOH water (0.86 cc)
(1.71 mmol) in water (5.6 cc) and stirred at room temperature for 14 hours. The precipitated solid was collected by filtration, washed with water and dried. This solid was dissolved in 5.8 cc of methanol and 10
% Pd-c 0.12 g was added, and the mixture was stirred under a hydrogen atmosphere for 1 hour. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue was subjected to CHP-20 column chromatography (H ₂ O → 5% C
H ₃ CN → 10% CH ₃ CN), and then (S, S) -4
-{2-Amino-3- (4-methoxyphenyl) propionyl} -3- (4-guanidinobutyl) -2-oxopiperazine-1-acetic acid was obtained. This intermediate (0.16g 0.3
6 mmol) and baking soda (0.09 g 1.07 mmol) in H ₂ O 3.2
cc, 1,4-dioxane 1.6 cc, and stirred at room temperature with 4-amidinobenzoyl chloride hydrochloride (0.10 g).
0.46 mmol) was added. After stirring for 1.5 hours, the reaction system was set to 1
The pH was adjusted to pH 4 with NH Cl water, and concentrated under reduced pressure. The residue was subjected to CHP-20 column chromatography (H ₂ O →
5% CH ₃ CN) to give the title compound (0.16 g, yield 27.3%) as a colorless amorphous powder. Specific rotation: [α] _D + 62.7 ° (C = 0.99, MeOH) Elemental analysis value: C ₂₉ H ₃₈ N ₈ O ₆ .HCl.3H ₂ O (685.
176) as calculated; C, 50.84; H, 6.62; N, 16.35 Found: C, 50.76; H, 6.47; N, 16.11

Reference Example 22 (S) -4- (4-amidinobenzoylamino) acetyl-3- {3- (4-guanidinobutanoylamino) propyl} -2-oxopiperazine-1-acetic acid hydrochloride (ie compound E) (S) -4-benzyloxycarbonylaminoacetyl-3- {3- (4-t-butoxycarbonylaminobutanoylamino) propyl} -2-oxopiperazine-1-acetic acid (0) obtained in Reference Example 16 0.33 g (0.56 mmol) was dissolved in 6.6 cc of trifluoroacetic acid, the mixture was stirred at room temperature for 1 hour, and the reaction system was concentrated under reduced pressure. Residue and baking soda (0.14
3.3 cc of an aqueous solution of g 1.68 mmol) was added to S-methylisothiourea sulfate (0.93 g 3.35 mmol) and 2N NaO.
It was added to 3.3 cc of an aqueous solution of H water (1.68 cc 3.35 mmol), and the mixture was stirred at room temperature for 14 hours. The reaction system is concentrated under reduced pressure,
The residue was subjected to CHP-20 column chromatography (H ₂ O →
5% CH ₃ CN → 10% CH ₃ CN → 15% CH ₃ CN), and then (S)-[4- (benzyloxycarbonylamino) -acetyl-3- {3- (4-guanidinobutyrylamino) ) -Propyl} -2-oxopiperazin-1-yl] -acetic acid was obtained. This intermediate was dissolved in 6.0 cc of methanol, 0.30 g of 10% Pd-C was added, and the mixture was stirred under a hydrogen atmosphere for 1 hour. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue and baking soda (0.19 g 2.23 mmol) were added to H ₂ O.
It was dissolved in 6.0 cc and 1,4-dioxane 3.0 cc and stirred at room temperature with 4-amidinobenzoyl chloride hydrochloride (0.1 cc).
6 g 0.73 mmol) was added. After stirring for 1.0 hour, the reaction system was adjusted to PH4 with 1N HCl water and concentrated under reduced pressure. The residue was subjected to CHP-20 column chromatography (H ₂
O) to give 0.09 g of the title compound (yield 24.7%).
Was obtained as a colorless amorphous powder. Specific rotation: [α] _D + 48.4 ° (C = 0.96, H ₂ O) Elemental analysis value: C ₂₄ H ₃₅ N ₉ O ₆・ 2HCL ・ 3.5H ₂ O (6
81.572) calculated: C, 42.29; H, 6.51; N, 18.50 Found: C, 42.34; H, 6.59; N, 18.28

Reference Example 23 4- (N-benzyloxycarbonyl) glycyl-1-t-butoxycarbonylmethyl-
2-oxopiperazine-3-acetic acid 4- (N-benzyloxycarbonyl) glycyl-1-t
-Butoxycarbonylmethyl-2-oxopiperazine-
1.46 g of 3-acetic acid methyl ester was dissolved in a mixed solution of 5 ml of water and 5 ml of methanol, and 190 mg of lithium hydroxide monohydrate was added at 0 ° C over 5 minutes. After stirring at the same temperature for 1 hour, the mixture was stirred at room temperature for 1 hour. The reaction solution was adjusted to pH 7 with 5% aqueous potassium hydrogen sulfate, and methanol was removed by concentration under reduced pressure. Further, the pH was adjusted to 3 with 5% potassium hydrogen sulfate, and then extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give the title compound (1.1 g) as a colorless oil. NMR: (CDCl ₃ ) δ: 1.452 (9H, s), 2.80-4.
65 (10H, m), 5.10 (2H, s), 5.82 (1H, m), 6.
03 (1H, m), 7.33 (5H, s). IR νmax, cm ^-1 : 3000,1730,1660,146
5,1370,1230,1160

Reference Example 24 3- (4-amidinophenyl)
Aminocarbonylmethyl-4- (N-benzyloxycarbonyl) glycyl-2-oxopiperazine-1-acetic acid
t-Butyl ester 4- (N-benzyloxycarbonyl) glycyl-1-t-butoxycarbonylmethyl-2-oxopiperazine-3-acetic acid 820 mg and 4-aminobenzamidine dihydrochloride 370 mg obtained in Reference Example 9 were added to pyridine. Dissolve in 5 ml, at room temperature 370 mg of dicyclohexylcarbodiimide, and 4
-Dimethylaminopyridine (10 mg) was added, and the mixture was stirred at room temperature for 24 hours. The insoluble material was filtered, the filtrate was concentrated under reduced pressure, and the crude product obtained was dissolved in 1% aqueous hydrochloric acid and subjected to column chromatography using CHP-20. 5% acetonitrile-
The fraction eluted with water is lyophilized to give 550 mg of the title compound.
Was obtained as a colorless powder. NMR (DMSOd _-6 ) δ: 1.42 (9H, s), 2.83-4.
44 (13H, m), 5.02 (2H, s), 7.34 (5H, s), 7.
78-7.82 (4H, m), 9.03-9.25 (3H, m). IR v max, cm ^-1 : 3325,1730,1680,16
40, 1480, 1365, 1260, 1155.

Reference Example 25 (S) -4- [N- (4-Amidinobenzoylamino) acetyl] -3- (4-amidinophenyl) aminocarbonylmethyl-2-oxopiperazine-1-acetic acid Obtained in Reference Example 12. 3- (4-amidinophenyl) aminocarbonylmethyl-4- (N-benzyloxycarbonyl)
Glycyl-2-oxopiperazine-1-acetic acid t-butyl ester (930 mg) was dissolved in methanol (15 ml) to give 10%.
Palladium-carbon (100 mg) was added, and the mixture was stirred at room temperature under a hydrogen atmosphere for 1 hour. The reduction catalyst was removed by filtration, the filtrate was concentrated under reduced pressure, and the oily substance obtained was 350 mg of sodium hydrogen carbonate.
Along with the solution dissolved in a mixed solution of 25 ml of water and 25 ml of dioxane, 307 mg of 4-amidinobenzoic acid was added over 5 minutes with vigorous stirring at room temperature. The crude product obtained by concentrating the reaction solution was dissolved in 5 ml of dichloromethane, 5 ml of trifluoroacetic acid was added at room temperature, and the mixture was stirred for 1 hour. The crude product obtained by concentrating the reaction solution under reduced pressure was purified by column chromatography using CHP-20 to obtain 490 mg of the title compound as a colorless powder. Specific rotation: [α] _D ²³ + 57.5 ° (C = 0.9, H ₂ O) Elemental analysis value: C ₂₅ H ₂₈ N ₈ O ₆ · CF ₃ CO ₂ H · 2.7H ₂
Calculated as O: C, 46.41; H, 4.96; N, 16.04 Found: C, 46.56; H, 4.80; N, 15.84

Reference Example 26 (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid hydrochloride (ie, compound A) ) (S) -4-benzyloxycarbonylaminoacetyl-3- (3-t-butoxycarbonylamino) obtained in Reference Example 2
0.7 g of propyl-2-oxopiperazine-1-acetic acid t-butyl ester was dissolved in 4.9 ml of trifluoroacetic acid and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and azeotropically distilled with toluene several times. The residue was subjected to column chromatography using CHP-20 (Mitsubishi Kasei) and the fraction eluted with 20% acetonitrile / water was concentrated (S) -4.
-Benzyloxycarbonylaminoacetyl-3- (3
-Amino) propyl-2-oxopiperazine-1-acetic acid was obtained as a crude product. This product was dissolved in 12.0 ml of methanol, 250 mg of 10% palladium-carbon was added, and the mixture was stirred under a hydrogen atmosphere for 1 hour. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue and 836 mg of the double layer were dissolved in a mixed solvent of 7.0 ml of 1,4-dioxane and 14.0 ml of water, and 4-guanidinobenzoic acid N-hydroxy-5-under stirring at room temperature.
1.27 mg of norbornene-2,3-dicarboxylic acid imide ester hydrochloride was added. After stirring for 1 hour, the reaction system was 1N.
The mixture was adjusted to pH 3-4 with hydrochloric acid and then concentrated under reduced pressure. The residue CHP-20 column chromatography (5% CH ₃ CN
/ Elute with H ₂ O) and freeze-dry the aliquot to give 0.48 g of the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 56.3 ° (C = 1.017, H ₂
O) Elemental analysis value: C ₂₇ H ₃₄ N ₁₀ O ₆・ 1.0 HCl ・ 3.5H ₂ O
Calculated as: C, 46.72; H, 6.10; N, 20.18 Found: C, 46.56; H, 6.17; N, 20.05

Reference Example 27 (S) -3- (2-t-butoxycarbonylamino) ethyl-2-oxopiperazine-1
- acid t- butyl ester oxalate (S) -N ² - benzyloxycarbonyl -N ^{4-t-butoxycarbonyl-2,4-diaminobutane} acid 26 g, N-
15.5 g of (2,2-dimethoxyethyl) glycine t-butyl ester was dissolved in 200 ml of acetonitrile, and 19 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added with stirring at room temperature. After stirring at the same temperature for additional 2 hours, the reaction solution was concentrated, the obtained oily substance was dissolved in ethyl acetate, washed with 5% aqueous potassium hydrogen sulfate solution and saturated aqueous sodium hydrogen carbonate solution, and dried over anhydrous magnesium sulfate. It was concentrated under reduced pressure. The residue was dissolved in 500 ml of toluene, 1.4 g of p-toluenesulfonic acid was added, and the mixture was stirred at 70 ° C. for 3 hours. The reaction solution cooled to room temperature was washed with saturated aqueous sodium hydrogen carbonate solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was dissolved in 500 ml of methanol, 10 g of 10% Pd-C was added, and the mixture was stirred under a hydrogen atmosphere at room temperature for 10 hours. The catalyst was removed by filtration, 6.4 g of oxalic acid was added to the filtrate, and the mixture was concentrated under reduced pressure. The obtained crude crystals were recrystallized from methanol / ethyl acetate to give the title compound (9.5 g) as colorless crystals. Mp: 165-169 ° C. Elemental _{analysis: C 17 H 31 N 3 O} 5 · (CO 2 H) 2 Calculated: C, 51.00; H, 7.43 ; N, 9.39 Found: C , 50.78; H, 7.59; N, 9.14

Reference Example 28 (S) -4- (benzyloxycarbonylamino) acetyl-3- (2-t-butoxycarbonylamino) ethyl-2-oxopiperazine-1-
Acetic acid t-butyl ester (S) -3- (2-t-butoxycarbonylaminoethyl) -2-oxopiperazine-1-acetic acid t obtained in Reference Example 27
900 mg of butyl ester oxalate was suspended in 20 ml of dichloromethane, 20 ml of saturated aqueous sodium hydrogencarbonate was added, and the mixture was vigorously stirred for 10 minutes. The organic layer was separated and dried over anhydrous magnesium sulfate, then 420 mg of N-benzyloxycarbonylglycine, and 1-ethyl-3- (3
-Dimethylaminopropyl) carbodiimide hydrochloride 50
0 mg was added and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, the residue was dissolved in ethyl acetate, washed with 5% potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate solution, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: ethyl acetate-hexane = 3: 1) to obtain 1.05 g of the title compound as a colorless oil. IR ν max cm ^-1 : 3450,1705,1655,1
640, 1500, 1450, 1360, 1240, 116
NMR (CDCl ₃ ) δ: 1.43 (9H, s), 1.46 (9H,
s), 2.05-2.33 (1H, m), 2.73-2.95 (1H,
m), 3.15-4.20 (10H, m), 5.05 (1H, dd, J =
3Hz), 5.13 (2H, s), 5.30 (1H, brs), 5.83
(1H, brs), 7.36 (5H, s).

Reference Example 29 (S) -4- (4-amidinobenzoylamino) acetyl-3- [2- (4-guanidinobenzoylamino)] ethyl-2-oxopiperazine-1
-Acetic acid hydrochloride (ie, compound D) (S) -4- (N-benzyloxycarbonylamino) acetyl-3- (2-t-butoxycarbonylamino) ethyl-2-oxopiperazine-1 obtained in Reference Example 28. -Acetic acid
550 mg of t-butyl ester was added to 5 ml of trifluoroacetic acid.
And was stirred at room temperature for 1 hour. The oily substance obtained by concentrating the reaction solution and 400 mg of sodium hydrogencarbonate were dissolved in a mixed solution of 25 ml of water and 25 ml of dioxane, and the mixture was stirred at room temperature.
250 mg of guanidinobenzoyl chloride hydrochloride were added. The reaction mixture was adjusted to pH 7 with 1N hydrochloric acid, 10% palladium-carbon (100 mg) was added, and the mixture was stirred under a hydrogen atmosphere for 1 hr. The catalyst is filtered off and 30 ml of dioxane is added to the filtrate.
And 400 mg of sodium hydrogen carbonate were added and 230 mg of 4-amidinobenzoic acid hydrochloride was added with vigorous stirring. The reaction mixture was adjusted to pH 3 with 1N hydrochloric acid and then concentrated under reduced pressure to a half volume. The concentrate was purified by CHP-20 column (5% acetonitrile / water) to give 250 mg of the title compound as a colorless amorphous solid. Specific rotation: [α] _D ²⁰ + 26.112 ° (C = 0.450,
MeOH) Elemental _{analysis: C 26 H 31 N 9 O} 6 · HCl · 5H 2 O Calculated: C, 45.12; H, 6.12 ; N, 18.21 Found: C, 45.61; H, 6.06; N, 18.22

Reference Example 30 (S) -4-benzyloxycarbonylaminoacetyl-3-t-butoxycarbonylaminomethyl-2-oxopiperazine-1-acetic acid t-
Butyl ester (S) -N ² -benzyloxycarbonyl-N ³ -t-butoxycarbonyl-2,3-diaminopropanoic acid was used to obtain the title compound as a colorless oil by the same method as in Reference Examples 1 and 2. It was H ¹ -NMR (CDCl ₃ ) δ, 1.38 (9H, s), 1.47 (9
H, s), 3.19-4.20 (10H, m), 4.90-5.05
(2H, m), 5.13 (2H, s), 5.82 (1H, brs), 7.36
(5H, s).

Reference Example 31 (S) -4- (4-amidinobenzoylamino) acetyl-3-aminomethyl-2-oxopiperazine-1-acetic acid dihydrochloride (S) -4-benzyl obtained in Reference Example 30 Oxycarbonylaminoacetyl-3-t-butoxycarbonylaminomethyl-2-oxypiperazine-1-acetic acid t-butyl ester was subjected to the same operations as in Reference Example 3 to obtain the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 44.9 ° (C = 0.655, Me
OH) Elemental _{analysis: C 17 H 22 N 6 O} 5 · 2HCl · 4H 2 O Calculated: C, 38.14; H, 6.02 ; N, 15.70 Found: C, 38.11; H, 5.65; N, 15.70

Reference Example 32 (S) -4- (4-amidinobenzoylamino) acetyl-3- (4-amidinobenzoylamino) methyl-2-oxopiperazine-1-acetic acid
Hydrochloride (S) -4- (4-amidinobenzoylamino) acetyl-3-aminomethyl-2-oxopiperazine-1-acetic acid dihydrochloride obtained in Reference Example 31 was subjected to the same operation as in Reference Example 4. , The title compound was obtained as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 60.2 ° (C = 0.535, MeO
H) Elemental analysis value: calculated as C ₂₅ H ₂₈ N ₈ O ₆ .HCl.3H ₂ O: C, 47.89; H, 5.63; N, 17.87 measured value: C, 47.63; H, 5.36; N, 17.81

Reference Example 33 (S) -4- (4-amidinobenzoylamino) acetyl-3- [2- (4-amidinobenzoylamino)] ethyl-2-oxopiperazine-1-
Acetic acid trifluoroacetate (S) -4- (benzyloxycarbonylamino) acetyl-3- (2-t-butoxycarbonylamino) ethyl-2-oxopiperazine-1-acetic acid t-butyl ester obtained in Reference Example 28 The title compound was obtained as a colorless amorphous powder by performing the same operations as in Reference Example 3 and Reference Example 4. Specific rotation: [α] D ²⁰ + 30.299 ° (C = 0.47)
0, H ₂ O) Elemental analysis value: C ₂₆ H ₃₀ N ₈ O ₆ .CF ₃ CO ₂ H. 3H ₂ O Calculated value: C, 46.80; H, 5.19; N, 15.59 actual measurement Value: C, 46.67; H, 4.99; N, 15.39

Reference Example 34 (S) -4- (4-guanidinobenzoylamino) acetyl-3- [2- (4-guanidinobenzoylamino)] ethyl-2-oxopiperazine-
1-Acetic acid trifluoroacetic acid salt (S) -4- (benzyloxycarbonylamino) acetyl-3- (2-t-butoxycarbonylamino) ethyl-2-oxopiperazine-1-acetic acid t-obtained in Reference Example 28
The title compound was obtained as a colorless amorphous powder by the same operation as in Reference Example 26 using butyl ester. Specific rotation: [α] _D ²⁰ + 35.207 ° (C = 0.650, H
₂ O) Elemental _{analysis: C 26 H 32 N 10 O} 6 · CF 3 CO 2 H · 3H 2 O
Calculated: C, 44.92; H, 5.25; N, 18.71 Found: C, 44.95; H, 5.54; N, 18.69

Reference Example 35 (R) -4- (4-amidinobenzoylamino) acetyl-3- (3-amino) propyl-2-oxopiperazine-1-acetic acid trifluoroacetic acid ZD-Orn (Boc)- OH was subjected to the same operations as in Reference Examples 1, 2 and 3 to obtain the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ −35.6 ° (C = 0.519, Me
OH) Elemental _{analysis: C 19 H 26 N 6 O} 5 · 2CF 3 CO 2 H · 1.5H
Calculated as ₂ O: C, 41.02; H, 4.64; N, 12.48 Measured value: C, 41.16; H, 4.47; N, 12.60

Reference Example 36 (R) -4- (4-amidinobenzoylamino) acetyl-3- [3- (4-amidinobenzoylamino)] propyl-2-oxopiperazine-1
-Acetic acid trifluoroacetic acid salt (R) -4- (4-amidinobenzoylamino) acetyl-3- (3-amino) propyl-2-oxopiperazine-1-acetic acid obtained in Reference Example 35 The title compound was obtained as a colorless amorphous powder by performing the same operation as in 4. Specific rotation: [α] _D ²⁰ −41.6 ° (C = 0.495, Me
OH) Elemental analysis value: C ₂₇ H ₃₂ N ₈ O ₆ .CF ₃ CO ₂ H. 4H ₂ O Calculated value: C, 46.40; H, 5.51; N, 14.93 Measured value: C, 46.66; H, 5.20; N, 14.90.

Reference Example 37 (S) -4- (4-amidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-
1-Acetic acid trifluoroacetic acid salt (S) -4- (benzyloxycarbonylamino) acetyl-3- (3-t-butoxycarbonylamino) propyl-2-oxopiperazine-1-acetic acid t-obtained in Reference Example 2
The butyl ester was subjected to the same operations as in Reference Example 29 to obtain the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 48.6 ° (C = 1.017, H
₂ O) Elemental _{analysis: C 27 H 33 N 9 O} 6 · 1.1CF 3 CO 2 H · 1.
Calculated as 5H ₂ O: C, 48.00; H, 5.30; N, 16.97 Found: C, 47.91; H, 5.11; N, 17.22

Reference Example 38 (S) -4- (4-amidinobenzoylamino) acetyl-3- (4-amidinophenylaminocarbonyl) ethyl-2-oxopiperazine-1
-Acetic acid trifluoroacetate (S) -4-benzyloxycarbonylaminoacetyl-
1-t-Butoxycarbonylmethyl-2-oxopiperazine-3-propanoic acid methyl ester was subjected to the same operations as in Reference Example 23, Reference Example 24, and Reference Example 25 to obtain the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 59.625 ° (C = 0.360,
H ₂ O) Elemental analysis value: C ₂₆ H ₃₀ N ₈ O ₆ .CF ₃ CO ₂ H. 4H ₂ O Calculated value: C, 45.65; H, 5.34; N, 15.21 Actual value: C, 45.70; H, 5.10; N, 14.91

Reference Example 39 (S) -4- (4-amidinobenzoylamino) acetyl-3- (4-amidinomethylbenzoylamino) methyl-2-oxopiperazine-1-
Acetic acid dihydrochloride (S) -4-benzyloxycarbonylaminoacetyl-3-t-butoxycarbonylaminomethyl-2-oxopiperazine-1-acetic acid t-butyl ester obtained in Reference Example 30 and 4-amidinomethylbenzoic acid The title compound was obtained as a colorless amorphous powder by subjecting N-hydroxysuccinimide active ester of hydrochloride and 4-amidinobenzoyl chloride hydrochloride to the same operation as in Reference Example 29. Specific rotation: [α] _D ²⁰ + 48.4 ° (C = 0.665, Me
OH) Elemental _{analysis: C 26 H 30 N 8 O} 6 · 2HCl · 4.5H 2 O Calculated: C, 44.32; H, 5.87 ; N, 15.90 Found: C, 44. 23; H, 5.74; N, 15.88

Reference Example 40 (S) -4- (4-amidinobenzoylamino) acetyl-3- (4-guanidinomethylbenzoylamino) methyl-2-oxopiperazine-1
-Acetic acid dihydrochloride (S) -4-benzyloxycarbonylaminoacetyl-3-t-butoxycarbonylaminomethyl-2-oxopiperazine-1-acetic acid t-butyl ester obtained in Reference Example 30 and 4-guanidinomethylbenzoic acid The title compound was obtained as a colorless amorphous powder by subjecting N-hydroxysuccinimide active ester of hydrochloride and 4-amidinobenzoyl chloride hydrochloride to the same operation as in Reference Example 29. Specific rotation: [α] _D ²⁰ + 47.2 ° (C = 0.553, H ₂
O) Elemental _{analysis: C 26 H 31 N 9 O} 6 · 2HCl · 3H 2 O Calculated: C, 45.09; H, 5.67 ; N, 18.20 Found: C, 45.32; H, 5.55; N, 18.10

Reference Example 41 (S, S) -4- [2- (4-amidinobenzoylamino) -3- (4-methoxyphenyl)] propionyl-3- [3- (6-aminohexanoylamino)] Propyl-2-oxopiperazine-1-acetic acid
Trifluoroacetate salt (S, S) -4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl)] propionyl-3- (3-t-butoxycarbonylamino) propyl-obtained in Reference Example 9. 2-oxopiperazine-1-acetic acid t-butyl ester was subjected to the same operations as in Reference Example 15 and Reference Example 16 to obtain the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 57.3 ° (C = 0.678, Me
OH) Elemental analysis value: C ₃₃ H ₄₅ N ₇ O ₇ · CF ₃ CO ₂ H · 2.5H ₂
Calculated as O: C, 51.85; H, 6.34; N, 12.09 Found: C, 52.02; H, 6.25; N, 12.04

Reference Example 42 (S, S) -4- [2- (4-amidinobenzoylamino) -3- (4-methoxyphenyl)] propionyl-3- [4- (2-aminoacetylamino)] butyl 2-Oxopiperazine-1-acetic acid trifluoroacetic acid salt (S, S) -4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl)] propionyl-3- (4 obtained in Reference Example 12 -T-butoxycarbonylamino) butyl-2-oxopiperazine-1-acetic acid t-butyl ester and Nt-butoxycarbonylglycine Reference Example 1
5 and Reference Example 16 were performed to obtain the title compound as a colorless amorphous powder. Specific rotation: [α] D ²⁰ + 59.8 ° (C = 0.644, Me
OH) Elemental _{analysis: C 30 H 39 N 7 O} 7 · CF 3 CO 2 H · 2.5H 2
Calculated as O: C, 50.00; H, 5.90; N, 12.75 Found: C, 49.95; H, 5.72; N, 12.87

Reference Example 43 4- (Amino-hydroxyimino) benzoic acid methyl ester 16.5 g of 4-cyanobenzoic acid methyl ester and 7.2 g of hydroxylamine hydrochloride were dissolved in 200 ml of methanol, and 8.82 g of sodium hydrogen carbonate at room temperature. After adding 3
Heated to reflux for an hour. After cooling, 400 ml of water was added and the precipitated crystals were collected by filtration. After washing the crystals with water and ether,
After drying under reduced pressure, 16.1 g of the title compound was obtained as colorless needle crystals. Mp: 170-172 ° C. Elemental analysis: Calculated as _{C 9 H 10 N 2 O 3} : C, 55.67; H, 5.19; N, 14.43 Found: C, 55.57; H, 5.22; N, 14.39

Reference Example 44 4- (2,5-dihydro-5-
(Oxo-1,2,4-oxadiazol-3-yl) benzoic acid 5.83 g of 4- (amino-hydroxyimino) benzoic acid methyl ester obtained in Reference Example 43 and 6 g of N, N′-carbonyldiimidazole were obtained. The suspension was suspended in 30 ml of dioxane and heated and stirred at 110 ° C for 30 minutes. The reaction solution was concentrated to dryness, the residue was dissolved in water and the pH was adjusted to 4 with acetic acid to precipitate crystals. The crystals were dissolved in 60 ml of 2N sodium hydroxide and stirred overnight at room temperature. Add acetic acid to the reaction mixture to add pH 4
The precipitated crystals were collected by filtration and washed with water. This crystal was recrystallized from dimethylformamide / ethyl acetate to give the title compound 4.
3 g was obtained as colorless crystals. Melting point: 300 ° C. or higher Elemental analysis value: C ₉ H ₆ N ₂ O ₄ Calculated value: C, 52.44; H, 2.93; H, 13.59 Measured value: C, 52.14: H, 3 .29; H, 13.89

Reference Example 45 (S) -4- [4- (2,5-dihydro-5-oxo-1,2,4-oxadiazole-3)
-Yl) benzoylamino] acetyl-3- {3- [4-
(2,5-Dihydro-5-oxo-1,2,4-oxadiazol-3-yl) benzoylamino]} propyl-2-
Oxopiperazine-1-acetic acid ammonium salt (S) -4- (benzyloxycarbonylamino) acetyl-3- (3-t-butoxycarbonylamino) propyl-2-oxopiperazine-1-acetic acid t obtained in Reference Example 2 −
Dissolve 1 g of butyl ester in 50 ml of methanol and
% Palladium-carbon (0.2 g) was added, and the mixture was stirred under a hydrogen stream for 1 hour. The catalyst was filtered off and the filtrate was concentrated under reduced pressure to give a residue, which was obtained in Reference Example 44 with 4- (2,5-dihydro-5-oxo-).
1,2,4-oxadiazol-3-yl) benzoic acid was added and dissolved in 20 ml of dimethylformamide, then 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (hereinafter referred to as WSC) 0 0.36 g was added and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography (eluted with ethyl acetate-25% methanol / ethyl acetate) to give an oil. This product was dissolved in 6 ml of trifluoroacetic acid, stirred at room temperature for 2 hours, and the reaction solution was concentrated under reduced pressure. The residue was dissolved in 8 ml of dimethylformamide, 1.25 ml of triethylamine was added, and then 4-
An active ester prepared from 0.33 g of (2,5-dihydro-5-oxo-1,2,4-oxadiazol-3-yl) benzoic acid, 0.23 g of N-hydroxysuccinimide and 0.42 g of dicyclohexylcarbodiimide. The dimethylformamide solution of was added and stirred at room temperature for 3 hours. The insoluble matter was removed by filtration, the filtrate was concentrated under reduced pressure, the residue was dissolved in water, and acetic acid was added to adjust to pH 4, and the precipitate was collected by filtration. This product was dissolved in water, adjusted to pH 8 with aqueous ammonia, applied to an XAD-2 column, and the fraction eluted with 10% acetonitrile / water was lyophilized to give 0.14 g of the title compound as a colorless amorphous powder. It was Specific rotation: [α] _D ²⁰ + 49.9 ° (C = 0.522, Me
OH) Elemental analysis value: calculated as C ₂₉ H ₃₁ N ₉ O ₁₀ · 3.5H ₂ O: C, 47.80; H, 5.26; N, 17.30 actual value: C, 47.87; H, 5.12; N, 17.81

Reference Example 46 4-Cyanobenzoic acid t-butyl ester 45.0 g of 4-cyanobenzoic acid and 3.1 ml of concentrated sulfuric acid were suspended in 612 ml of methylene chloride, 310 ml of isobutene was added with stirring at 0 ° C., and the mixture was stirred at room temperature. Stir for 13 days. The reaction solution was neutralized with saturated multi-layered water, and then extracted and extracted with ethyl acetate,
The organic layer was concentrated under reduced pressure. The precipitated solid was collected by filtration and washed with hexane. The filtrate and washings are combined and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography (hexane / ethyl acetate = 10/1) and crystallized from methylene chloride / petroleum ether to give the title compound (43.1 g) as white crystals. NMR (CDCl ₃ ) δ: 1.61 (9H, s), 7.72 (2H, d,
J = 8.8Hz), 8.08 (2H, d, J = 8.8Hz)

Reference Example 47 4- (Amino-hydroxyimino) methyl-benzoic acid t-butyl ester 4.3 Cyanobenzoic acid t-butyl ester 4.3 g, hydroxylamine hydrochloride 1.84 g, sodium hydrogencarbonate 2.3
1 g was dissolved in a mixed solvent of 21.2 ml of t-butanol and 2.1 ml of water and stirred at 80 ° C. for 2 hours. Water was added to the reaction solution, the mixture was extracted with ethyl acetate, and the organic layer was concentrated under reduced pressure.
The residue was purified by silica gel column chromatography (hexane / ethyl acetate = 1/1) and crystallized from hexane to give the title compound (4.41 g) as colorless needle crystals. Mp: 153-155 ° C. Elemental _{analysis: C 12 H 16 N 2 O} 3 Calculated: C, 61.00; H, 6.83 ; N, 11.86 Found: C, 61.03; H, 6.70; N, 11.90

Reference Example 48 4- (Amino-methoxycarbonyloxyiminomethyl) benzoic acid t-butyl ester 4- (amino-hydroxyimino) methyl-benzoic acid t-
Butyl ester 1.0 g, potassium carbonate 292 mg 1,4
-Dissolved in 8.46 ml of dioxane and under stirring at 0 ° C 343 μl of methyl chloroformate was added. After stirring at room temperature for 1 hour, water was added to the reaction solution. The precipitated crystals were collected by filtration and washed with water to obtain 1.22 g of the title compound as white crystals. Mp: 157-159 ° C. Elemental _{analysis: C 14 H 18 N 2 O} 5 Calculated: C, 57.14; H, 6.16 ; N, 9.52 Found: C, 56.98; H, 6.21; N, 9.30

Reference Example 49 4- (amino-methoxycarbonyloxyiminomethyl) benzoic acid trifluoroacetic acid salt 4- (amino-methoxycarbonyloxyiminomethyl)
1.0 g of benzoic acid t-butyl ester was dissolved in 4.0 ml of trifluoroacetic acid and stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and azeotroped with toluene to give the title compound (0.80 g) as a colorless amorphous powder. Elemental _{analysis: C 10 H 10 N 2 O} 5 · CF 3 CO 2 H (352.2
233) as calculated: C, 40.92; H, 3.15; N, 7.95 Found: C, 41.21; H, 2.98; N, 7.96

Reference Example 50 (S) -4- [4- (Amino-methoxycarbonyloxyiminomethyl) benzoylamino] acetyl-3- {3- [4- (amino-methoxycarbonyloxyiminomethyl) benzoylamino]} Propyl-2-oxopiperazine-1-acetic acid (S) -4- (benzyloxycarbonylamino) acetyl-3- (3-t-butoxycarbonylamino) propyl-2-oxopiperazine-1-obtained in Reference Example 2 Acetic acid t−
Butyl ester and 4- (amino-obtained in Reference Example 20)
Methoxycarbonyloxyiminomethyl) benzoic acid Trifluoroacetic acid salt was used and subjected to the same operations as in Reference Example 26 to obtain the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 50.5 ° (C = 1.018, Me
OH) Elemental _{analysis: C 31 H 36 N 8 O} 12 · 2H 2 O Calculated: C, 49.73; H, 5.38 ; N, 14.97 Found: C, 49.54; H, 5.19; N, 14.87

Reference Example 51 (S) -4- (4-amidinobenzoylamino) acetyl-3- {3- [4- (amino-methoxycarbonyloxyiminomethyl) benzoylamino]} propyl-2-oxopiperazine-1 -Acetic acid hydrochloride (S) -4- (benzyloxycarbonylamino) acetyl-3- (3-t-butoxycarbonylamino) propyl-2-oxopiperazine-1-acetic acid t-obtained in Reference Example 2
The butyl ester, 4- (amino-methoxycarbonyloxyiminomethyl) benzoic acid trifluoroacetic acid salt and 4-amidinobenzoic acid obtained in Reference Example 49 were subjected to the same operations as in Reference Example 29 to give the title compound as a colorless amorphous compound. Obtained as a powder. Specific rotation: [α] _D ²⁰ + 47.5 ° (C = 1.00, H ₂ O) Elemental analysis value: Calculated as C ₂₉ H ₃₄ N ₈ O ₉ · HCl · 3H ₂ O: C, 47. 77; H, 5.67; N, 15.37 Found: C, 47.51; H, 5.68; N, 15.27.

Reference Example 52 (S) -3- [3- (4-amidinobenzoylamino)] propyl-4-benzyloxycarbonylaminoacetyl-2-oxopiperazine-1-
Acetic acid 1.35 g of (S) -4-benzyloxycarbonylaminoacetyl-3- (3-t-butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid t-butyl ester obtained in Reference Example 2 was trifluoro. It was dissolved in 6.8 ml of acetic acid, stirred at room temperature for 1 hour, and concentrated under reduced pressure. The residue was dissolved in a mixed solvent of 20 ml of water and 10 ml of dioxane,
At room temperature, 806 mg of sodium hydrogen carbonate and then 683 mg of 4-amidinobenzoyl chloride hydrochloride were added, and the mixture was vigorously stirred for 30 minutes. The crude product obtained by concentrating the reaction solution is CH
Purification by P-20 column gave 1.41 g of the title compound as a colorless amorphous powder. Elemental _{analysis: C 27 H 32 N 6 O} 7 · 2H 2 O Calculated: C, 55.09; H, 6.16 ; N, 14.28 Found: C, 55.36; H, 6 . 10; N, 14.35

Reference Example 53 (S) -4- [4- (2-Aminoethyl) benzoylamino] acetyl-3- [3- (4-amidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid tri Fluoroacetate (that is, compound C) (S) -3- [3- (4-amidinobenzoylamino)] propyl-4-benzyloxycarbonylaminoacetyl-2-oxopiperazine-1-acetic acid 30 obtained in Reference Example 52
0 mg was dissolved in 20 ml of methanol and 10% palladium-
Carbon (120 mg) was added, and the mixture was stirred under a hydrogen stream at room temperature for 1 hr.
The catalyst was filtered off and the filtrate was concentrated under reduced pressure to give an oily substance which was dissolved in 5 ml of dimethylformamide and dissolved in 173 mg of 4- (2-t-butoxycarbonylamino) benzoic acid and 1-ethyl-3- (3- Dimethylaminopropyl) carbodiimide hydrochloride (167 mg) was added, and the mixture was stirred at room temperature for 2 hours. The oily substance obtained by concentrating the reaction solution was dissolved in 7 ml of trifluoroacetic acid and stirred at room temperature for 1 hour. The crude product obtained by concentrating the reaction solution under reduced pressure was used as a CHP-20 column (5% acetonitrile /
(Elution with water) to obtain 110 mg of the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 41.7 ° (C = 1.018, Me
OH) Elemental analysis value: C ₂₈ H ₃₅ N ₇ O ₆ · 1.1CF ₃ CO ₂ H · 4
Calculated as H ₂ O: C, 47.53; H, 5.82; N, 12.85 Found: C, 47.64; H, 5.60; N, 12.72

Reference Example 54 (S) -4- (4-amidinobenzoylamino) acetyl-3- [3- (4-amidinobenzoylamino)] propyl-2-oxopiperazine-1
-Acetic acid hydrochloride (S) -4- (4-amidinobenzoylamino) acetyl-3- [3- (4-amidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid trifluoro obtained in Reference Example 4 Dissolve 1 g of acetate salt in 5 ml of 0.5 N hydrochloric acid,
After stirring at C for 5 minutes, it was adsorbed on a CHP-20 column and washed with water until the eluate became neutral. Next, the fraction eluted with 10% acetonitrile / water was freeze-dried to obtain 0.7 g of the title compound as a colorless amorphous powder. Specific rotation: + 51.3 ° (C = 1.018, H ₂ O) Elemental analysis value: C ₂₇ H ₃₂ N ₈ O ₆ · HCl · 5H ₂ O Calculated value: C, 46.92; H, 6 .27; N, 16.21 Found: C, 47.13; H, 6.14; N, 16.23.

Reference Example 55 N- (4-t-Butoxycarbonylphenyl) -N'-ethoxycarbonylthiourea 4-aminobenzoic acid ^t- butyl ester 13.51 g was dissolved in isopropyl ether 150 ml and stirred at room temperature with ethoxycarbonyl. Isothiocyanate 9.83g
Was added. After stirring for 2 hours, the precipitated crystals were collected by filtration,
Recrystallize from isopropyl ether to give the title compound 21.
83 g were obtained as colorless needle crystals. Mp: 119-120 ° C. Elemental _{analysis:. C 15 H 20 N 2} O 4 S Calculated C, 55.54; H, 6.21; N, 8.64 Found C, 55.56; H, 6 .06; N, 8.65

Reference Example 56 N- (4-butoxycarbonylphenyl) -N'-ethoxycarbonyl-S-methylisothiourea N- (4-t-butoxycarbonylphenyl) -N'-ethoxycarbonyl obtained in Reference Example 55 Thiourea 21.7
g was dissolved in 80 ml of tetrahydrofuran, 2.68 g of 60% oily sodium hydride was washed with hexane and then added in an ice bath with stirring. Next, 9.5 g of methyl iodide
A 30 ml solution of hexane was added dropwise under the same conditions. After completion of dropping, the mixture was stirred for 1 hour under the same conditions. The reaction solution is concentrated under reduced pressure,
The residue was dissolved in ethyl acetate, washed with water, and concentrated under reduced pressure. The residue was recrystallized from hexane to obtain 20 g of the title compound as colorless needle crystals. Mp: 67-68 ° C. Elemental analysis _{C 16 H 22 N 2 O 4} S Calculated C, 56.78; H, 6.55; N, 8.28 Found C, 56.63; H, 6.31 ; N, 8.15

Reference Example 57 3- (4-t-butoxycarbonylphenylamino) -1,2,4-oxadiazolin-4H-5-one N- (4-butoxycarbonylphenyl) -obtained in Reference Example 56 22.8 g of N'-ethoxycarbonyl-S-methylisothiourea and 14 g of hydroxylamine hydrochloride were dissolved in 350 ml of methanol, 18 g of triethylamine was added dropwise while stirring in an ice bath, and the mixture was further stirred at room temperature for 14 hours. The reaction solution was concentrated under reduced pressure, the residue was dissolved in ethyl acetate and washed with 1N hydrochloric acid. The crude product obtained by concentrating the organic layer under reduced pressure was recrystallized from ethyl acetate-hexane to obtain 7.8 g of the title compound as colorless prism crystals. Melting point: 271-272 ° C. (decomposition) Elemental analysis Calculated as C ₁₃ H ₁₅ N ₃ O ₄ 1 / 10H ₂ O C, 55.95; H, 5.49; N, 15.06 Measured value C, 55 .81; H, 5.47; N, 15.05

Reference Example 58 3- (4-Carboxyphenylamino) -1,2,4-oxazoline-4H-5-one 3- (4-t-butoxycarbonylphenylamino) -1, obtained in Reference Example 57, 2,4-oxadiazoline-4H-
5.7 g of 5-one was dissolved in 70 ml of 1N sodium hydroxide and stirred at 115 ° C. for 1.5 hours. After cooling, the reaction solution was neutralized with 2N hydrochloric acid, and the deposited precipitate was extracted with ethyl acetate. The crude crystals obtained by concentrating the extract under reduced pressure were washed with ethyl acetate to obtain 5.36 g of the title compound as yellow crystals. Mp: 272-273 ℃ (decomposition) Elemental analysis C ₉ H ₇ N ₃ O ₄ Calculated: C, 47.58; H, 3.40 ; N, 18.50 Found: C, 47.76; H , 3.39; N, 18.57.

Reference Example 59 4-Carboxyphenyl Cyanamide 4-Amino (N-hydroxyimino) methyl benzoic acid methyl ester 17.12 g was added to tetrahydrofuran 180
It was dissolved in ml, 12.12 g of triethylamine was added, and 12.65 g of methanesulfonyl chloride was added dropwise in an ice bath. The mixture was stirred under the same conditions for 1 hour and concentrated under reduced pressure. Methanol was added to the residue, and the precipitated crystals were collected by filtration. This product was dissolved in 100 ml of methanol, and 100 ml of water containing 12 g of sodium hydroxide was added with stirring at room temperature. Methanol was distilled off under reduced pressure, 700 ml of water was added to the residue, and 80 ml of 4N hydrochloric acid was added with stirring at room temperature. The precipitated crystals were collected by filtration to give the title compound (13.12 g) as colorless crystals. Melting point: 300 ° C. or higher Elemental analysis: Calculated as C ₈ H ₆ N ₂ O ₂ C, 59.26; H, 3.73; N, 17.28 Found C, 58.97; H, 3.82; N, 17.04

Reference Example 60 N- (4-Carboxyphenyl) -N'-hydroxyguanidine 6.56 g of 4-carboxyphenylcyanamide obtained in Reference Example 59 was dissolved in 150 ml of methanol, and hydroxylamine hydrochloride 6 was added with stirring at room temperature. 0.1 g and triethylamine 8.88 g were added and stirred for 2 hours. The precipitated crystals were collected by filtration to give the title compound (4.45 g) as colorless crystals. Mp: 200-202 ° C. (decomposition) Elemental _{analysis: C 8 H 9 N 3 O} 3 Calculated C, 48.78; H, 4.71; N, 21.33 Found C, 48.55; H, 4.69; N, 21.09

Reference Example 61 3- (4-Carboxyphenylamino) -5-trifluoromethyl-1,2,4-oxadiazole N- (4-carboxyphenyl) -obtained in Reference Example 60
4.0 g of N'-hydroxyguanidine was dissolved in 100 ml of tetrahydrofuran, and 6.75 g of trifluoroacetic anhydride was added while stirring at 0 ° C. The mixture was stirred under the same conditions for 1.5 hours and concentrated under reduced pressure. Water was added to the residue and the precipitated crystals were collected by filtration and recrystallized from ethyl acetate-hexane to give the title compound (3.5 g) as colorless crystals. Mp: 244-246 ° C. Elemental _{analysis: C 10 H 6 N 3 O} 3 Calculated _{F 3 C, 43.97; H,} 2.21; N, 15.38 Found C, 44.06; H, 2 .31; N, 15.28

Reference Example 62 4-t-Butoxycarbonylbenzaldoxime 4-cyanobenzoic acid ^t- butyl ester 20.5 g and hydroxime amine hydrochloride 13.9 g were dissolved in methanol 100 ml and triethylamine 128 g was added with stirring at room temperature. Stir at 1 ° C for 1 hour. The reaction solution was concentrated under reduced pressure, the residue was dissolved in ethyl acetate and washed with water. The crude product obtained by concentrating the organic layer under reduced pressure was recrystallized from isopropyl ether to obtain 11.45 g of the title compound as colorless crystals. Melting point: 113-114 ° C. Elemental analysis: Calculated as C ₁₂ H ₁₆ N ₂ O ₃ .1 / 10H ₂ O C, 60.54; H, 6.86; N, 11.77 Measured value C, 60.77 H, 6.79; N, 11.57

Reference Example 63 4-t-butoxycarbonylphenylcyanamide 16.3 g of 4-t-butoxycarbonylbenzaldoxime obtained in Reference Example 62 was dissolved in 150 ml of ethyl acetate, and 13.7 ml of triethylamine was added to the mixture in an ice bath. 9.92 g of methanesulfonyl chloride was added dropwise. After stirring under the same conditions for 0.5 hours, the reaction solution was washed with water and the organic layer was depressurized. The residual oil was dissolved in 150 ml of tetrahydrofuran and stirred at room temperature with 2N sodium hydroxide 75
ml was added and the mixture was stirred for 0.5 hours. After that, tetrahydrofuran was distilled off under reduced pressure. The residual liquid was neutralized with 2N hydrochloric acid, extracted with ethyl acetate, and concentrated under reduced pressure. The residue was recrystallized from hexane-isopropyl ether to give the title compound as colorless crystals. Mp: 94-95 ° C. Elemental analysis _{C 12 H 14 N 2 O 2} · 1 / 10H 2 O Calculated C, 65.50; H, 6.50; N, 12.73 Found C, 65.51; H, 6.51; N, 12.52

Reference Example 64 N- (4-t-butoxycarbonylphenyl) -N'-methoxycarbonyloxyguanidine 8.72 g of 4-t-butoxycarbonylphenylcyanamide obtained in Reference Example 63 and hydroxylamine hydrochloride 5.56
g was dissolved in 120 ml of methanol, and 8.80 g of triethylamine was added dropwise at -25 ° C. After completion of the dropping, the reaction solution was brought to room temperature and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and washed with water, and then 3.16 g of pyridine and 3.78 g of methyl chlorocarbonate were added to the organic layer at -10 ° C. After the temperature of the reaction solution was set to room temperature, it was washed with water and the organic layer was concentrated under reduced pressure. The crude product was recrystallized from isopropyl ether to give the title compound 9.
39 g was obtained as colorless crystals. Mp: 122-126 ° C. Elemental analysis C ₁₄ H ₁₉ N ₃ O ₅ Calculated C, 54.36; H, 6.19; H, 13.58 Found C, 54.29; H, 6.02; H, 13.41

Reference Example 65 N- (4-carboxyphenyl) -N'-methoxycarbonyloxyguanidine 9.2 g of N- (4-t-butoxycarbonylphenyl) -N'-methoxycarbonyloxyguanidine obtained in Reference Example 64 Is dissolved in 25 ml of trifluoroacetic acid and the mixture is allowed to stand at room temperature for 2
Stirred for hours. 100 ml of water was added to the residue obtained by concentrating the reaction solution under reduced pressure, and sodium hydrogencarbonate was added to adjust the pH to 6
The crude crystal that had been adjusted to the above was collected by filtration. This product was recrystallized from tetrahydrofuran-ethyl acetate to give the title compound 4.5.
3 g was obtained as colorless crystals. Mp: 174-175 ° C. Elemental analysis C ₁₀ H ₁₁ N ₃ O ₅ Calculated C, 47.43; H, 4.38; N, 16.59 Found C, 47.17; H, 4.33; N, 16.44

Reference Example 66 (S) -2-oxo-4-
[4- (5-Oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoyl] aminoacetyl-3- [4- (5-oxo-4,5-dihydro [1 ,
2,4] Oxadiazol-3-ylamino) benzoyl] aminopropylpiperazine-1-acetic acid (S) -4-benzyloxycarbonylaminoacetyl-3- (3- ^t -butoxycarbonylaminopropyl) obtained in Reference Example 2 500 mg of 2-oxopiperazine-1-acetic acid ^t- butyl ester was dissolved in 5 ml of trifluoroacetic acid and stirred at room temperature for 1 hour. The oily substance obtained by concentrating the reaction solution under reduced pressure was dissolved in 10 ml of methanol, 10 mg of 10% palladium-carbon was added, and the mixture was stirred under hydrogen at room temperature for 1 hour. The catalyst was filtered off and the filtrate was concentrated under reduced pressure (S)-
A crude product of 4-aminoacetyl-3-aminopropyl-2-oxopiperazine-1-acetic acid was obtained. This product is water 10
It was dissolved in 10 ml of dioxane and 400 mg of sodium hydrogen carbonate was added. Next, 3-obtained in Reference Example 58
(4-Carboxyphenylamino) -1,2,4-oxadiazolin-4H-5-one (420 mg) and N-hydroxysuccinimide (250 mg) were dissolved in dimethylformamide (5 ml) to prepare dicyclohexylcarbodiimide 450.
After adding mg, the mixture was stirred at room temperature for 3 hours and then concentrated under reduced pressure to obtain an oily substance. This product was dissolved in 5 ml of dioxane, added to the solution of (S) -4-aminoacetyl-3-aminopropyl-2-oxopiperazine-1-acetic acid prepared above, and stirred at room temperature for 6 hours. The reaction mixture was neutralized with 1N hydrochloric acid and then concentrated under reduced pressure to obtain a crude product, which was Sephadex LH-.
Purification by 20 columns gave 230 mg of the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ 51.9 ° (C = 0.27, DMS
O) Elemental analysis _{C 29 H 30 N 10 O 10} · H 2 O Calculated: C, 50.00; H, 4.63 ; N, 20.11 Found: C, 49.79; H, 4 . 91; N, 19.96.

Reference Example 67 (S) -2-oxo-4-
[4- (5-Trifluoromethyl [1,2,4] -oxadiazol-3-ylamino) benzoyl] aminoacetyl-3- [4- (5-trifluoromethyl [1,2,4]-
Oxadiazol-3-ylamino) benzoyl] propylpiperazine-1-acetic acid (S) -4-benzyloxycarbonylaminoacetyl-3- (3- ^t -butoxycarbonylaminopropyl) -2-oxopiperazine obtained in Reference Example 2 -1-Acetic acid ^t- butyl ester and 3- (4-carboxyphenylamino) -5-trifluoromethyl-1,2,4 obtained in Reference Example 61
By a method similar to that in Reference Example 66 using -oxadiazole, the title compound was obtained as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ 39.7 ° (C = 0.25, DMS
O) Elemental analysis: Calculated as C ₃₁ H ₂₈ N ₁₀ O ₈ F ₆ · H ₂ O: C, 46.51; H, 3.78; N, 17.49 Found: C, 46.44; H , 3.97; N, 17.26.

Reference Example 68 (S) -4- [4- (N-methoxycarbonyloxyguanidino) benzoylaminoacetyl] -3- [3- (N-methoxycarbonyloxyguanidino) benzoylamino] propyl-2-oxopiperazine -1-Acetic acid (S) -4-benzyloxycarbonylaminoacetyl-3- (3- ^t -butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid ^t- butyl ester obtained in Reference Example 2 and Reference Example 65 The title compound was obtained as a colorless amorphous powder in the same manner as in Reference Example 66 using the obtained N- (4-carboxyphenyl) -N'-methoxycarbonyloxyguanidine. Specific rotation: [α] _D ²⁰ 31.20 ° (C = 0.28, DM
SO) Elemental _{analysis: C 31 H 38 N 10 O} 12 · 2H 2 O Calculated: C, 47.81; H, 5.44 ; N, 17.99 Found: C, 47.63; H, 5 .71; N, 17.83

Reference Example 69 (S) -4- (N- ^t -butoxycarbonylamino) acetyl-3- (3- ^t -butoxycarbonylamino) propyl-2-oxopiperazine-
1-Acetic acid (S) -4-benzyloxycarbonylaminoacetyl-3- (3- ^t -butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid ^t- butyl ester (1.5 g) obtained in Reference Example 2 was trifluoro It was dissolved in 10 ml of acetic acid and stirred at room temperature for 1 hour. The oil and the reaction solution was concentrated under reduced pressure to give dissolved in a mixture of H ₂ O10ml and dioxane 10 ml, sodium bicarbonate 400mg and di ^t
700 mg of butyl dicarbonate was added, and the mixture was stirred at room temperature for 3 hours. Dioxane was distilled off under reduced pressure, the resulting aqueous solution was washed with ethyl acetate, and potassium hydrogen sulfate was added to adjust the pH.
Adjusted to 3.5 and extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give the title compound (1.2 g) as colorless crystals. Melting point: 107-109 ° Elemental analysis: Calculated as C ₂₁ H ₃₆ N ₄ O ₈ : C, 53.38; H, 7.68; N, 11.86 Found: C, 53.35; H, 7 .73; N, 11.95.

Reference Example 70 (S) -4- (N-benzyloxycarbonylamino) acetyl-3- (3-benzyloxycarbonylamino) propyl-2-oxopiperazine-1-acetic acid Obtained in Reference Example 2 (S ) -4-Benzyloxycarbonylaminoacetyl-3- (3- ^t -butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid ^t- butyl ester (2.0 g) was dissolved in trifluoroacetic acid (10 ml) and stirred at room temperature for 1 hour. did. The oily substance obtained by concentrating the reaction solution under reduced pressure was dissolved in a mixed solution of 10 ml of H ₂ O and 10 ml of dioxane, 600 mg of sodium hydrogen carbonate and 550 mg of carbobenzoxylchloride were added, and the mixture was stirred at room temperature for 1 hour. Dioxane was distilled off under reduced pressure, the obtained aqueous solution was washed with ethyl acetate, potassium hydrogen sulfate was added to adjust the pH to 3.5, and the mixture was extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give the title compound (1.5 g) as a colorless amorphous powder. Elemental analysis: Calculated as C ₂₇ H ₃₂ N ₄ O ₈ : C, 59.99; H, 5.97; N, 10.36 Found: C, 60.13; H, 5.87; N, 10 .22.

Reference Example 71 (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid pivaloyloxymethyl ester Two
Hydrochloride (S) -4- (N-benzyloxycarbonylamino) acetyl-3- (3-benzyloxycarbonylamino) propyl-2-oxopiperazine-obtained in Reference Example 70
1-Acetic acid (500 mg), potassium carbonate (128 mg) and potassium iodide (463 mg) were dissolved in dimethylformamide (5 ml), and pivaloyloxymethyl chloride 420 was dissolved at room temperature.
mg was added and the mixture was stirred at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure, the obtained oily substance was dissolved in ethyl acetate, washed with 10% potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate, and then concentrated under reduced pressure. The residue is dissolved in 10 ml of methanol and 10%
Palladium-carbon (100 mg) was added, the mixture was stirred under hydrogen for 1 hr, the catalyst was filtered off, and the filtrate was concentrated. The obtained oily substance was dissolved in a mixed solution of 20 ml of water and 20 ml of dioxane, and 400 mg of sodium hydrogen carbonate and 3,5-dioxo-4-azatricyclo [5,5-guanidinobenzoic acid]
750 mg of 2,1,0 2,6] dec-8-en-4-yl ester hydrochloride was added, and the mixture was stirred at room temperature for 3 hours. After dioxane was distilled off under reduced pressure, 1N. Hydrochloric acid was added to the resulting aqueous solution to adjust the pH to 3.5, and the product was purified by CHP-20 column to obtain 240 mg of the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ 56.23 ° (C = 0.27, H ₂
O) Elemental analysis: Calculated as C ₃₃ H ₄₄ N ₁₀ O ₈ .2HCl.H ₂ O: C, 49.56; H, 6.05; N, 17.51 Found: C, 49.31; H , 6.33; N, 17.24

Reference Example 72 (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid 1- (cyclohexyloxycarbonyl) (Oxy) ethyl ester dihydrochloride (S) -4- (N-benzyloxycarbonylamino) acetyl-3- (3-benzyloxycarbonylamino) propyl-2-oxopiperazine-obtained in Reference Example 70
The title compound was obtained as a colorless amorphous powder by the same operation as in Reference Example 71 using 1-acetic acid and 1- (cyclohexyloxycarbonyloxy) ethyl chloride. Specific rotation: [α] _D ²⁰ 52.5 ° (C = 0.50, H ₂
O) Elemental analysis: Calculated as C ₃₆ H ₄₈ N ₁₀ O ₉ · 2HCl · 3H ₂ O: C, 48.49; H, 6.33; N, 15.71 Found: C, 48.35; H , 6.33; N, 15.52.

Reference Example 73 (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid 5-methyl-2- Oxo-1,3-
Dioxolen-4-ylmethyl ester dihydrochloride (S) -4- (N- ^t -butoxycarbonylamino) acetyl-3- (3- ^t -butoxycarbonylamino) propyl-2-oxopiperazine-1 obtained in Reference Example 69 -Acetic acid 300 mg and sodium hydrogencarbonate 62 mg were dissolved in dimethylformamide 5 ml to give 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl bromide 1
15 mg was added and the mixture was stirred at room temperature for 5 hours. The reaction solution was concentrated under reduced pressure, the obtained oily substance was dissolved in ethyl acetate, washed with 10% potassium hydrogen sulfate and saturated aqueous sodium hydrogen carbonate, and then concentrated under reduced pressure. The residue was dissolved in 5 ml of trifluoroacetic acid and stirred at room temperature for 1 hour, and then concentrated under reduced pressure to obtain an oily substance, which was dissolved in a mixed solution of 20 ml of water and 20 ml of dioxane,
400 mg of sodium hydrogen carbonate and 4-guanidinobenzoic acid 3,5-dioxo-4-azatricyclo [5,2,
500 mg of 1,0 2,6] dec-8-en-4-yl ester hydrochloride was added, and the mixture was stirred at room temperature for 3 hours. Dioxane was distilled off under reduced pressure, and 1N hydrochloric acid was added to the resulting aqueous solution to obtain p.
After adjusting to H3.5, the product was purified by CHP-20 column to obtain 115 mg of the title compound as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ 43.7 ° (C = 1.0, MeO
H) Elemental analysis: Calculated as C ₃₂ H ₃₈ N ₁₀ O ₉ · 2HCl · 3H ₂ O: C, 46.10; H, 5.56; N, 16.80 Found: C, 46.43; H , 5.41; N, 16.58

Reference Example 74 (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid 2- (isobutyloxycarbonyl) ) -2-Propyridene ethyl ester dihydrochloride (S) -4- (N- ^t -butoxycarbonylamino) acetyl-3- (3- ^t -butoxycarbonylamino) propyl-2-oxopiperazine-obtained in Reference Example 69 The title compound was obtained as a colorless amorphous powder by the same procedure as in Reference Example 73 using 1-acetic acid and 2- (isobutyloxycarbonyl) -2-propylideneethyl bromide. Specific rotation: [α] _D ²⁰ 47.34 ° (C = 0.48, H ₂
O) Elemental analysis: C ₃₇ H ₅₀ N ₁₀ O ₈・ 2CF ₃ CO ₂ H ・ 2H ₂
Calculated as O: C, 47.95; H, 5.50; N, 13.64. Found: C, 48.05; H, 5.51; N, 13.54.

Reference Example 75 (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid ethyl ester dihydrochloride Reference Reference example using (S) -4- (N- ^t -butoxycarbonylamino) acetyl-3- (3- ^t -butoxycarbonylamino) propyl-2-oxopiperazine-1-acetic acid and ethyl iodide obtained in Example 69. The title compound was obtained as a colorless amorphous powder by the same operation as in 73. Specific rotation: [α] _D ²⁰ 49.30 ° (C = 0.47, H ₂
O) Elemental analysis: Calculated as C ₂₉ H ₃₈ N ₁₀ O ₆ .2HCl.2H ₂ O: C, 47.61; H, 6.06; N, 19.14 Found: C, 47.29; H , 6.35; N, 18.88

Reference Example 76 (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3-tert-butoxycarbonylaminopropyl) -2-oxo Piperazine-1
-Yl] acetic acid tert-butyl ester (Also known as: (S,
S) -4- [2-Benzyloxycarbonylamino-3
-(4-Methoxyphenyl) propionyl] -3- (3
-Tert-butoxycarbonylaminopropyl) -2-oxopiperazine-1-acetic acid tert-butyl ester) (S)-[3- (3-tert-butoxycarbonylamino) propyl-2-oxopiperazine obtained in Reference Example 3 -1
-Yl] acetic acid tert-butyl ester / oxalate 4.
2 g was dissolved in 50 ml of water, 2.3 g of NaHCO ₃ was added and 50 ml of dichloromethane was extracted twice and dried (Na ₂ S
After O ₄ ) it was concentrated under reduced pressure. Z-Tyr (OM
e) -OH (3 g) was added and the mixture was dissolved in dichloromethane (150 ml). Then, WSC (1.92 g) was added and the mixture was stirred at room temperature for 2 hours. After dichloromethane was distilled off under reduced pressure, it was extracted with ethyl acetate. The ethyl acetate phase was washed with 3% KHSO ₄ water and saturated NaHCO ₃ water, dried (Na ₂ SO ₄ ), and concentrated under reduced pressure. The residue was purified by silica gel chromatography (hexane / AcOEt = 1: 2 to AcOEt) to obtain 5.88 g of the title compound. ¹ H NMR (CDCl ₃ ) δ 1.35 to 2.10 (4
H, m), 1.41 (9H, s), 1.46 (9H, s), 2.3
0 (1H, m), 2.80 to 3.85 (7H, m), 3.41
(1H, d, J = 17.4Hz), 3.78 (3H, s), 4.
24 (1H, d, J = 17.4Hz), 4.75 (2H,
m), 4.94 (1H, t, J = 6.5Hz), 5.10 (2
H, q, J = 12.4 Hz), 5.69 (1H, d, J = 8.2)
Hz), 6.80 (2H, d, J = 8.6Hz), 7.09
(2H, d, J = 8.6Hz), 7.35 (5H, s).

Reference Example 77 (S, S)-[3- (3-Aminopropyl) -4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl]
2-Oxopiperazin-1-yl] acetic acid (Also known as:
(S, S) -3- (3-aminopropyl) -4- [2-
Benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -2-oxopiperazine-1
-Acetic acid) (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) obtained in Reference Example 76.
Propionyl] -3- (3-tert-butoxycarbonylaminopropyl) -2-oxopiperazin-1-yl]
Acetic acid tert-butyl ester 5.7 g toluene 20 ml
Suspended in water, stirred under ice cooling, then trifluoroacetic acid 20 m
1 was added. After stirring at room temperature for 2 hours, toluene was added and the mixture was concentrated under reduced pressure. The residue was dissolved in 30 ml of water, conc. The pH was adjusted to 5 with aqueous ammonia and then purified by XAD-2 column chromatography (eluted with H ₂ O → 50% CH ₃ CN water) to obtain 4.3 g of the title compound. ¹ H NMR (CD ₃ OD) δ 1.40 to 2.10 (4
H, m), 2.32 (1H, m), 2.80 to 4.00 (7H,
m), 3.16 (1H, d, J = 16.5Hz), 3.77
(3H, s), 4.61 to 4.85 (2H, m), 4.72
(1H, d, J = 16.5Hz), 5.05 (2H, q, J =
12.3 Hz), 6.82 (2H, d, J = 8.4 Hz), 7.
11 (2H, d, J = 8.4 Hz), 7.32 (5H, s).

Reference Example 78 (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazine -1
-Yl] acetic acid (also known as (S, S) -4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazine -1-Acetic acid) (S, S)-[3- (3-aminopropyl) -4- [2-benzyloxycarbonylamino-obtained in Reference Example 77.
3- (4-Methoxyphenyl) propionyl] -2-
Oxopiperazin-1-yl] acetic acid (3.8 g) was dissolved in 50% dioxane water (100 ml) to obtain NaHCO _{3 (} 1.52 g).
After the addition of 1., 1.24 ml of Z-chloride was added dropwise under ice cooling, and the mixture was further stirred at room temperature for 1.5 hours. Dioxane was distilled off, 3% KHSO ₄ water was added to adjust the pH to 2, and the mixture was extracted with ethyl acetate, washed with saturated NaHCO ₃ water, and dried (Na ₂ SO 4
After ₄ ), it was concentrated under reduced pressure. Ether was added to the residue and decantation was performed twice to obtain 4 g of the title compound. ¹ H NMR (CDCl ₃ ) δ 1.40 to 2.05 (4
H.m), 2.22 (1H, m), 2.75 (9H, m), 3.
74 (3H, s), 4.65-5.20 (6H, m), 5.5
2 (1H, t, J = 5.5Hz), 5.94 (1H, d, J =
8.6 Hz), 6.78 (2H, d, J = 8.6 Hz), 7.0
5 (2H, d, J = 8.6Hz), 7.31 (10H, s).

Reference Example 79 (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazine -1
-Yl] acetic acid tert-butyl ester (Alternative name:
(S, S) -4- [2-Benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3
-(3-benzyloxycarbonylaminopropyl)-
2-oxopiperazine-1-acetic acid tert-butyl ester) (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl)) obtained in Reference Example 78.
Propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazin-1-yl]
After 1.7 g of acetic acid, 2 ml of tert-butanol and 1.6 g of 4-dimethylaminopyridine were dissolved in 50 ml of dichloromethane, 0.6 g of WSC was added and the mixture was stirred at room temperature for 24 hours. Dichloromethane was distilled off, and the mixture was extracted with ethyl acetate. The ethyl acetate phase was washed with water and saturated aqueous NaCl, dried (Na ₂ SO ₄ ), and concentrated under reduced pressure. The residue was purified by silica gel chromatography (AcOEt) and crystallized from ether / hexane to give 1.02 g of the title compound as colorless crystals. mp.138-140 ° C. Specific rotation: [α] _D ²⁰ + 49.7 ° (C = 0.431, MeO)
H) Elemental analysis value: C ₃₉ H ₄₈ N ₄ O ₉ (716.832), calculated value: C, 65.35; H, 6.75; N, 7.82 analysis value: C, 65.17; H, 6.69; N, 7.91

Reference Example 80 (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- [3- (4-guanidinobenzoylamino) propyl] -2 -Oxopiperazin-1-yl] acetic acid hydrochloride (Also known as: (S, S) -4-
[2-Benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- [3- (4-guanidinobenzoylamino) propyl] -2-oxopiperazine-1-acetic acid hydrochloride) Obtained in Reference Example 78. (S, S)-[3- (3-aminopropyl) -4- [2-benzyloxycarbonylamino-
0.5 g of 3- (4-methoxyphenyl) propionyl] -2-oxopiperazin-1-yl]] acetic acid was dissolved in 50 ml of 50% aqueous dioxane and NaHCO ₃ 0.24 g.
After addition of 4-guanidinobenzoic acid 3,5-dioxo-4-aza-tricyclo [5,2,1,0 2,6] deca-
0.377 g of 8-en-4-yl ester hydrochloride was added, and the mixture was stirred at room temperature for 2 hours. Adjust the pH of the reaction mixture to 1N-HC
After diluting to 3 with l, dioxane is distilled off. XAD the residue
-2 column chromatography (H ₂ O → 10% CH ₃ C
(N water → 20% CH ₃ CN water → 50% CH ₃ CN water elution)
Purification by to give 0.43 g of the title compound. ¹ H-NMR (CD ₃ OD) δ: 1.50 to 2.10 (4
H, m), 2.45 (1H, m), 2.80 to 4.25 (9H,
m), 3.77 (3H, s), 4.60 to 5.00 (2H,
m), 5.00 (2H, s), 6.83 (2H, d, J = 8.5)
Hz), 7.13 (2H, d, J = 8.5Hz), 7.30
(5H, s), 7.35 (2H, d, J = 8.5Hz), 7.9
2 (2H, d, J = 8.5 Hz).

Reference Example 81 (S, S)-[3- [3-
(4-guanidinobenzoylamino) propyl] -4-
[3- (4-Methoxyphenyl) -2- [4- (5-trifluoromethyl- [1,2,4] oxadiazole-3
-Ylamino) benzoylamino] propionyl] -2
-Oxopiperazin-1-yl] acetic acid / hydrochloride (Also known as:
(S, S) -3- [3- (4-guanidinobenzoylamino) propyl] -4- [3- (4-methoxyphenyl) -2- [4- (5-trifluoromethyl- [1,2,
4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxopiperazine-1-acetic acid / hydrochloride) (S, S)-[4- [2-benzyloxycarbonylamino] obtained in Reference Example 80 -3- (4-methoxyphenyl)
Propionyl] -3- [3- (4-guanidinobenzoylamino) propyl] -2-oxopiperazin-1-yl] acetic acid hydrochloride was dissolved in 40 ml of methanol to give 10
% Pd-C (0.2 g) was added, and catalytic reduction was performed at room temperature for 2 hours. After removing the catalyst by filtration and concentrating the filtrate under reduced pressure, 50
It was dissolved in 50 ml of% dioxane water. While keeping alkaline with NaHCO ₃ , 4- (5-trifluoromethyl-
A dioxane solution of acid chloride prepared from [1,2,4] oxadiazol-3-ylamino) benzoic acid and oxalyl chloride was added dropwise, and the mixture was stirred at room temperature for 30 minutes. The pH of the reaction solution was adjusted to 3 with 1N-HCl and then concentrated to dryness under reduced pressure. The residue was subjected to silica gel chromatography (AcOE
It was purified by t: AcOH: H ₂ O = 8: 1: 1) and ether was added to obtain 0.17 g of the title compound as a colorless powder. Specific rotation: [α] _D ²⁰ + 42.8 ° (C = 0.94, DMS
O). Elemental _{analysis: C 37 H 39 N 10 O} 8 F 3 · HCl · 0.1Et 2
Calculated as O (852.649): C, 52.68; H, 5.08; N, 16.43 Analytical value: C, 52.62; H, 5.01; N, 16.58

Reference Example 82 (S, S)-[4- [3-
(4-Methoxyphenyl) -2- [4- (5-trifluoromethyl [1,2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo-3- [3-[- 4- (5-trifluoromethyl [1,
2,4] oxadiazol-3-ylaminobenzoylamino] propyl] piperazin-1-yl] acetic acid tert
-Butyl ester (Also known as: (S, S) -4- [3- (4
-Methoxyphenyl) -2- [4- (5-trifluoromethyl [1,2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo-
3- [3- [4- (5-trifluoromethyl [1,2,
4] Oxadiazol-3-ylaminobenzoylamino] propyl] piperazine-1-acetic acid tert-butyl ester) (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl)
Propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazin-1-yl]
Acetic acid tert-butyl ester 0.54 g was added to methanol 5
It was dissolved in 0 ml and 10% Pd-C (0.25 g) was added, and catalytic reduction was carried out for 2 hours. The catalyst was filtered off, and the filtrate was concentrated to dryness under reduced pressure. 4- (5-trifluoromethyl-) in the residue
[1,2,4] Oxadiazol-3-ylamino) benzoic acid 0.41 g and 4-dimethylaminopyridine 0.1 g
Dissolved in 30 ml of acetonitrile, then WSC
0.39 g was added and the mixture was stirred for 20 hours. Acetonitrile was distilled off, extracted with ethyl acetate, washed with 3% KHSO ₄ water and saturated NaCl water, dried (Na ₂ SO ₄ ), and concentrated under reduced pressure. The residue was chromatographed on silica gel (Ac
Purification by OEt) gave 0.44 g of the title compound. ¹ H NMR (CD ₃ OD) δ: 1.45 (9 H, s), 1.
50-2.10 (4H, m), 2.47 (1H, m), 2.9
5 to 4.20 (9H, m), 3.77 (3H, s), 4.80
~ 5.20 (2H, m), 6.86 (2H, d, J = 8.6H
z), 7.20 (2H, d, J = 8.6Hz), 7.41 (2
H, d, J = 8.8Hz), 7.42 (2H, d, J = 8.8H)
z), 7.72 (2H, d, J = 8.8Hz), 7.77 (2
H, d, J = 8.8 Hz).

Reference Example 83 (S, S)-[4- [3-
(4-Methoxyphenyl) -2- [4- (5-trifluoromethyl [1,2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo-3- [3-[- 4- (5-trifluoromethyl [1,
2,4] oxadiazol-3-ylaminobenzoylamino] propyl] piperazin-1-yl] acetic acid (Also known as (S, S) -4- [3- (4-methoxyphenyl))
-2- [4- (5-trifluoromethyl [1,2,4] oxadiazol-3-ylamino) benzoylamino]
Propionyl] -2-oxo-3- [3- [4- (5-
Trifluoromethyl [1,2,4] oxadiazole-3
-Ylaminobenzoylamino] propyl] piperazine-1-acetic acid) (S, S)-[4- [3- (4-methoxyphenyl) -2- [4- (5-trifluoromethyl) obtained in Reference Example 82. [1,2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo-3- [3
-[-4- (5-Trifluoromethyl [1,2,4] oxadiazol-3-ylaminobenzoylamino] propyl] piperazin-1-yl] acetic acid tert-butyl ester 0.44 g under ice-cooling tri-ester. 6 ml of fluoroacetic acid was added and dissolved, and the mixture was stirred at room temperature for 2 hours.Toluene was added to the reaction solution, and the mixture was concentrated to dryness twice under reduced pressure, and then dissolved in a small amount of ethyl acetate.
I got Specific rotation: [α] _D ²⁰ + 0.7 ° (C = 1.043, DMS
O) Elemental _{analysis: C 39 H 36 N 10 O} 9 F 6 · 2H 2 O · 0.2Ac
Calculated as OEt (956.419): C, 49.98; H, 4.38; N, 14.64 Analytical value: C, 50.17; H, 4.17; N, 14.35

Reference Example 84 (S, S)-[4- [3-
(4-Methoxyphenyl) -2- [4- (5-oxo-
4,5-dihydro [1,2,4] oxadiazole-3-
Ilamino) benzoylamino] propionyl] -2-
Oxo-3- [4- (5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoylamino] propyl] piperazin-1-yl] acetic acid
tert-Butyl ester (Alternative name: (S, S) -4- [3
-(4-Methoxyphenyl) -2- [4- (5-oxo-4,5-dihydro [1,2,4] oxadiazole-3
-Ylamino) benzoylamino] propionyl] -2
-Oxo-3- [4- (5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoylamino] propyl] piperazine-1-acetic acid tert
-Butyl ester) (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) obtained in Reference Example 79.
Propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazin-1-yl]
Acetic acid tert-butyl ester 0.54 g was added to methanol 5
It was dissolved in 0 ml and 10% Pd-C (0.25 g) was added, and catalytic reduction was carried out for 2 hours. The catalyst was filtered off, and the filtrate was concentrated to dryness under reduced pressure. To the residue was added 4- (5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoic acid (0.33 g), which was dissolved in N, N-dimethylformamide (10 ml) and iced. Stir under cold. Then, 0.63 ml of triethylamine was added, 0.25 g of diethyl cyanophosphate was added, and the mixture was stirred under ice cooling for 1 hour. N, N-
Dimethylformamide was distilled off under reduced pressure, water was added, and 3% K
The mixture was adjusted to pH 2 with HSO ₄ water, extracted with ethyl acetate containing a small amount of N, N-dimethylformamide, washed with saturated aqueous NaCl, dried (Na ₂ SO ₄ ), and concentrated under reduced pressure. The residue was subjected to silica gel chromatography (AcOEt → Ac
Purification with OEt / AcOH / H ₂ O = 8: 1: 1) gave 0.52 g of the title compound. ¹ H NMR (CD ₃ OD) δ: 1.44 (9H, s),
1.50-2.10 (4H, m), 2.48 (1H, m), 2.
90 to 4.20 (9H, m), 3.75 (3H, s), 4.8
0 to 5.20 (2H, m), 6.84 (2H, d, J = 8.4
Hz), 7.18 (2H, d, J = 8.4Hz), 7.33
(2H, d, J = 8.8Hz), 7.35 (2, Hd, J = 8.8.
8Hz), 7.71 (2H, d, J = 8.8Hz), 7.74
(2H, d, J = 8.8Hz).

Reference Example 85 (S, S)-[4- [3-
(4-Methoxyphenyl) -2- [4- (5-oxo-
4,5-dihydro [1,2,4] oxadiazole-3-
Ilamino) benzoylamino] propionyl] -2-
Oxo-3- [4- (5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoylamino] propyl] piperazin-1-yl] acetic acid (also known as (S, S) -4- [3- (4-methoxyphenyl) -2- [4- (5-oxo-4,5-dihydro [1,
2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo-3- [4-
(5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoylamino] propyl] piperazine-1-acetic acid) (S, S)-[4 -[3- (4-Methoxyphenyl) -2- [4- (5-oxo-4,5-dihydro [1,2,4] oxadiazol-3-ylamino) benzoylamino] propionyl] -2-oxo −
3- [4- (5-oxo-4,5-dihydro [1,2,
4] Oxadiazol-3-ylamino) benzoylamino] propyl] piperazin-1-yl] acetic acid tert-butyl ester (0.62 g) was added to trifluoroacetic acid (6 m) under ice cooling.
1 was added and dissolved, and the mixture was stirred at room temperature for 2 hours. Toluene was added to the reaction mixture, and the mixture was concentrated to dryness twice under reduced pressure, dissolved in a small amount of methanol, and ethyl acetate was added to give 0.43 g of the title compound as a powder. Specific rotation: [α] _D ²⁰ + 7.5 ° (C = 0.983, DMS
O) Elemental analysis value: C ₃₇ H ₃₈ N ₁₀ O ₁₁ · 1.5H ₂ O · 0.5A
Calculated as cOEt (869.846): C, 53.85; H, 5.21; N, 16.10 Analytical value: C, 53.71; H, 5.05; N, 15.97

Reference Example 86 (S, S)-[4- [2-
[4- (3-methoxycarbonyloxyguanidino) be
Nazoylamino-3- (4-methoxyphenyl) propyi
Onyl] -3- [3- [4- (3-methoxycarbonyl
Oxyguanidino) benzoyl] amino] propyl-2
-Oxopiperazin-1-yl] acetic acid tert-butyl ether
Stell (Also known as: (S, S) -4- [2- [4- (3-me
Toxycarbonyloxyguanidino) benzoylamino
-3- (4-Methoxyphenyl) propionyl] -3-
[3- [4- (3-methoxycarbonyloxyguanidinium
No) Benzoyl] amino] propyl-2-oxopipera
Zin-1-acetic acid tert-butyl ester) (S, S)-[4- [2-benzyl obtained in Reference Example 79
Luoxycarbonylamino-3- (4-methoxyphenyl)
Lu) propionyl] -3- (3-benzyloxycarbo)
Nylaminopropyl) -2-oxopiperazine-1-i
] Acetic acid tert-butyl ester 0.5 g in methanol
Dissolve in 50 ml, add 0.25 g of 10% Pd-C, and add 2.
The time catalytic reduction was performed. The catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure.
It was dried to dryness. 4- (3-methoxycarbonylo) was added to the residue.
Xyguanidino) benzoic acid (0.38 g) was added, and N, N-di
Dissolve in 10 ml of methylformamide and stir under ice cooling.
Was. Then, after adding 0.21 ml of triethylamine
Add 0.25 g of diethyl cyanophosphate and under ice cooling at 1:00
It was stirred for a while. Acetic acid (1 ml) was added and the mixture was evaporated under reduced pressure. Residual
Silica gel chromatography (AcOEt → Ac
OEt / AcOH / H Purify with O = 18: 1: 1)
0.51 g of the title compound was obtained.¹ 1 H NMR (CD_ThreeOD) δ: 1.44 (9H, s), 1.
50-2.10 (4H, m), 2.48 (1H, m), 2.9
0-4.20 (9H, m), 3.76 (3H, s), 3.84
(6H, s), 4.80 to 5.20 (2H, m), 6.84
(2H, d, J = 8.6Hz), 7.18 (2H, d, J = 8.
6Hz), 7.33 (4H, d, J = 8.6Hz), 7.67
(4H, d, J = 8.6Hz).

Reference Example 87 (S, S)-[4- [2-
[4- (3-Methoxycarbonyloxyguanidino) benzoylamino-3- (4-methoxyphenyl) propionyl] -3- [3- [4- (3-methoxycarbonyloxyguanidino) benzoyl] amino] propyl-2
-Oxopiperazin-1-yl] acetic acid (Also known as: (S,
S) -4- [2- [4- (3-Methoxycarbonyloxyguanidino) benzoylamino-3- (4-methoxyphenyl) propionyl] -3- [3- [4- (3-methoxycarbonyloxyguanidino) benzoyl ] Amino] propyl-2-oxopiperazine-1-acetic acid) (S, S)-[4- [2- [4-
(3-Methoxycarbonyloxyguanidino) benzoylamino-3- (4-methoxyphenyl) propionyl] -3- [3- [4- (3-methoxycarbonyloxyguanidino) benzoyl] amino] propyl-2-oxopiperazine-1 -Yl] acetic acid tert-butyl ester (0.51 g) was dissolved by adding 6 ml of trifluoroacetic acid under ice cooling, and the mixture was stirred at room temperature for 2 hours. Toluene was added to the reaction solution and the mixture was concentrated to dryness twice under reduced pressure. The residue was dissolved in 50% methanol water and CHP-20 column chromatography (H ₂ O → 20% methanol water → 50% methanol water →
(75% aqueous methanol) to give 0.2 g of the title compound as a powder. Specific rotation: [α] _D ²⁰ + 9.7 ° (C = 1.04, DMSO) Elemental analysis value: C ₃₉ H ₄₆ N ₁₀ O ₁₃ · 0.5H ₂ O (871.
862), calculated value: C, 53.73; H, 5.43; N, 16.07 Analytical value: C, 53.76; H, 5.46; N, 16.09

Reference Example 88 (S, S) -4- [2- (4
-Guanidinobenzoyl) amino-3- (4-methoxyphenyl) propionyl] -3- [3- (4-guanidinobenzoyl) aminopropyl] -2-oxopiperazin-1-yl] acetic acid hydrochloride Obtained in Reference Example 77 (S, S) -3- (3-aminopropyl) -4- [2-benzyloxycarbonylamino-3
250 mg of-(4-methoxyphenyl) propionyl] -2-oxopiperazin-1-yl] acetic acid was dissolved in 5 ml of methanol, 100 mg of 10% palladium-carbon was added, and the mixture was stirred under hydrogen at room temperature for 1 hour. The catalyst was removed by filtration, and the oily substance obtained by concentrating the filtrate under reduced pressure was added to dioxane 10
dissolved in a mixed solvent of 10 ml of water and 10 ml of water, 210 mg of sodium hydrogen carbonate and 4-guanidinobenzoic acid 3,5
-Dioxo-4-azatricyclo [5,2,1,0 2,6]
Add 450 mg deca-8-en-4-yl ester,
Stirred at room temperature for 1 hour. The reaction solution was adjusted to pH 3 with 1N hydrochloric acid, and then dioxane was distilled off under reduced pressure. The obtained aqueous solution was applied to a CHP-20 column, and the fraction eluted with 10% acetonitrile / water was lyophilized to give the title compound 130.
mg was obtained as an amorphous powder. Elemental _{analysis: C 35 H 42 N 10 O} 7 · HCl · 2H 2 O Calculated: C, 53.40; H, 6.02 , N, 17.79 Found: C, 53.11; H, 5 .86, N, 18.06

Reference Example 89 (S)-[3- [3- (4-Amidinobenzoylamino) propyl] -4-[[4- (iminomethoxycarbonylaminomethyl) benzoylamino] acetyl] -2-oxopiperazine- 1-yl] acetic acid hydrochloride (Also known as: (S) -3- [3- (4-amidinobenzoylamino) propyl] -4-[[4- (iminomethoxycarbonylaminomethyl) benzoylamino] acetyl] -2- Oxopiperazine-1-acetic acid hydrochloride) (S)-[4-[(4-amidinobenzoylamino) acetyl] -3- [3- (4-amidinobenzoylamino) propyl] -2-obtained in Reference Example 4. Oxopiperazine-1-
Il] acetic acid hydrochloride (0.17 g, 0.29 mmol) in 1,4-dioxane (2.0 ml) and H ₂ O (2.0 m)
1), and 2N aqueous sodium hydroxide solution (0.46 ml, 0.91 mmol) was slowly added under ice cooling, and then chlorocarbonic acid methyl ester (0.053 ml, 0.6) was added.
9 mmol) was added and the mixture was stirred for 30 minutes. The reaction solution was adjusted to pH 3 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure. The residue was subjected to column chromatography (CHP-20, H ₂ O-5% C
_{H 3 CNaq-10% CH 3} CNaq-15% CH 3 CNaq)
The product was purified by the method described above, and converted to a hydrochloride with a 1N aqueous hydrochloric acid solution to obtain the title compound (0.20 g, 91%) as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 49.7 ° (C = 0.984%)
in MeOH (in methanol)) Elemental analysis value: C ₂₉ H ₃₄ N ₈ O ₈ 2.0 HCl 2.5 H ₂
Calculated as O.1.0 MeOH (772.640): C, 46.64; H, 5.87; N, 14.50 Analytical value: C, 46.34; H, 5.62; N, 14. 26

Reference Example 90 (S, S)-[4- [2- [4-
(3-Methoxycarbonylguanidino) benzoylamino] -3- (4-methoxyphenyl) propionyl] -3
-[3- [4- (3-Methoxycarbonylguanidino) benzoylamino] propyl] -2-oxopiperazine-1
-Yl] acetic acid hydrochloride (Also known as: (S, S) -4- [2-
[4- (3-Methoxycarbonylguanidino) benzoylamino] -3- (4-methoxyphenyl) propionyl] -3- [3- [4- (3-methoxycarbonylguanidino) benzoylamino] propyl] -2-oxopiperazine -1-Acetic acid hydrochloride) (S, S)-[4- [2- (4-guanidinobenzoylamino) -3- (4-methoxyphenyl) propionyl] -3- [3- (obtained in Reference Example 88) 4-guanidinobenzoylamino) propyl] -2-oxopiperazine-1-
Il] acetic acid (0.52 g, 0.73 mmol) in 1,4-
It was dissolved in dioxane (5.2 ml) and H ₂ O (5.2 ml), and a 2N sodium hydroxide aqueous solution (4.3
5 ml, 8.70 mmol) and chlorocarbonic acid methyl ester (0.55 ml, 7.25 mmol) were added while keeping the pH of the reaction system at 10 or less, and the mixture was stirred for 30 minutes. The reaction solution was adjusted to pH 7 with a 1N aqueous hydrochloric acid solution and then concentrated under reduced pressure. The residue was dissolved in H ₂ O (5.0 ml), and lithium hydroxide (0.20 g, 4.78 mmol) was added under ice cooling. After stirring at 0 ° C. for 2 hours, the reaction system was adjusted to pH 3 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure. The residue was purified by column chromatography _{(CHP-20,10% CH 3 CNaq} -1
5% CH ₃ CNaq-20% CH ₃ CNaq-25% CH ₃ C
Naq) and purified by _{(LH-20, H 2 O} ), to give the title compound (0.28g, 39%) as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 64.4 ° (C = 1.041%
in MeOH) Elemental analysis value: C ₃₉ H ₄₆ N ₁₀ O ₁₁・ 2.0HCl ・ 4.5H
Calculated as ₂ O (984.845): C, 47.56; H, 5.83; N, 14.22 Analytical value: C, 47.40; H, 5.55; N, 14.33

Reference Example 91 (S, S)-[4- [2-Benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazine -1-
Iyl] acetic acid 1-cyclohexyloxycarbonyloxyethyl ester (S, S)-[4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3-) obtained in Reference Example 78. Benzyloxycarbonylaminopropyl) -2-oxopiperazin-1-yl] acetic acid (0.58 g, 0.88 mmol) and triethylamine (0.49 ml, 3.52 mmol) in DMF (5.8 m).
l) and dissolved at room temperature under stirring with carbonic acid
1-Chloroethyl ester Cyclohexyl ester (0.73 g, 3.52 mmol) and potassium iodide (0.58 g, 3.52 mmol) were added. After stirring for 38 hours at room temperature, the reaction system was poured into water, and liquid separation extraction was performed with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (hexane / ethyl acetate = 2 /
Purification in 5) gave the title compound (0.43 g, 59%) as a colorless amorphous powder. IR νmax cm ^{- 1:} 3410,2930,175
5,1710,1645,1510,1450,1240,1
075 NMR (CDCl ₃ ) δ: 1.10-2.10 (14H,
m), 1.52 (3H, d, J = 5.4Hz), 2.80-5.
20 (15H, m), 3.77 (3H, s), 5.07 (2
H, s), 5.09 (2H, s), 5.64 (1H, d, J =
7.8 Hz), 6.67-6.87 (2H, m), 7.08
(2H, d, J = 8.4Hz), 7.33 (10H, s)

Reference Example 92 (S, S)-[4- [2- (4
-Guanidinobenzoylamino) -3- (4-methoxyphenyl) propionyl] -3- [3- (4-guanidinobenzoylamino) propyl] -2-oxopiperazin-1-yl] acetic acid 1-cyclohexyloxycarbonyloxyethyl ester (S, S)-[4- [2-Benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3-benzyloxycarbonylaminopropyl) -2-oxopiperazine obtained in Reference Example 91 -1-yl] acetic acid 1-cyclohexyloxycarbonyloxyethyl ester (0.43 g, 0.52 mmol) and acetic acid (0.062 ml, 1.09 mmol) in methanol (8.
6 ml). 10% Pd-C in this solution
(0.17 g) was added, and the mixture was stirred under a hydrogen atmosphere for 1 hour.
The catalyst was filtered off and the filtrate was concentrated under reduced pressure. 1 residue
4-dioxane (4.3 ml) and H ₂ O (8.6 ml)
Dissolved in water and stirred at room temperature with baking soda (0.22 g, 2.59 m).
mol) and 4-guanidinobenzoic acid N-hydroxy-5-norbornene-2,3-dicarboximide ester (0.43 g, 1.14 mmol) were added. After 1 hour, the reaction system was adjusted to pH 3 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure. The residue is subjected to column chromatography (CHP
_{-20,10% CH 3 CNaq-15%} CH 3 CNaq-20
% CH ₃ CNaq) and purified by _{(LH-20, H 2 O} ), to give the title compound (0.073g, 14%) as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 63.4 ° (C = 1.09%
in MeOH) Elemental _{analysis: C 44 H 56 N 10 O} 10 · 2.0HCl · 3.0H
Calculated as ₂ O (1011.957): C, 52.22; H, 6.37; N, 13.84 Analytical value: C, 52.38; H, 6.07; N, 13.81

Reference Example 93 (S, S)-[3- (3- ^t -Butoxycarbonylaminopropyl) -4- [2- [4-
(1,3-Dimethoxycarbonylguanidino) benzoylamino] -3- (4-methoxyphenyl) propionyl] -2-oxopiperazin-1-yl] acetic acid ^t- butyl ester (S, S)-[obtained in Reference Example 9 4- [2-benzyloxycarbonylamino-3- (4-methoxyphenyl) propionyl] -3- (3- ^t -butoxycarbonylaminopropyl) -2-oxopiperazin-1-yl] acetic acid ^t- butyl ester (0.66 g , 0.97 mmol) was dissolved in methanol (6.6 ml) and 10% Pd-C (0.26) was added.
g) was added, and the mixture was stirred under a hydrogen atmosphere for 1 hour. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue was dissolved in 1,4-dioxane (6.6 ml) and H ₂ O (6.6 ml), and 4-guanidinobenzoic acid N-hydroxy-5-norbornene-2,3-dicarboximide ester (0.55 g) was added. , 1.45 mmol) and baking soda (0.12 g,
1.45 mmol) was added at room temperature. After 1 hour, the reaction system was adjusted to pH 3 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure. The residue was subjected to column chromatography (CHP-20,
_{H 2 0-5% CH 3 CNaq-} 10% CH 3 CNaq-15%
_{CH 3 CNaq-25% CH 3} CNaq-30% CH 3 CNa
q) and purified by (S, S)-[3- (3- ^t -butoxycarbonylaminopropyl) -4- [2- (4-guanidinobenzoylamino) -3- (4-methoxyphenyl).
Propionyl] -2-oxopiperazin-1-yl] acetic acid
^t- Butyl ester (0.50 g, 73%) was obtained as a colorless amorphous powder. This product is 1,4-dioxane (5.0m
l) and stirred at 0 ° C., 2N aqueous sodium hydroxide solution (2.46 ml, 4.93 mmol) and chlorocarbonic acid methyl ester (0.27 ml, 3.52 mmol) were added while keeping the pH of the reaction system at 10 or less. added. 3 at 0 ° C
After stirring for 0 minutes, the reaction system was adjusted to pH 3 with a 1N aqueous hydrochloric acid solution, and separated and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and then concentrated under reduced pressure.
The residue was purified by silica gel column chromatography (hexane / ethyl acetate = 1/10) to give the title compound (0.42 g, 72%) as a colorless amorphous powder. IR νmax cm ^-1 (KBr): 3400,297
0,1730,1640,1510,1490,1435,1
362, 1245, 1155, 1025, 948 NMR (CD ₃ OD) δ: 1.39 (9H, s), 1.46.
(9H, s), 1.20-1.65 (2H, m), 1.65-
2.06 (2H, m), 2.41-2.64 (1H, m), 2.
88-4.18 (7H, m), 3.46 (3H, s), 3.5
9 (1H, d, J = 17.2Hz), 3.72 (3H, s),
3.77 (3H, s), 4.08 (1H, d, J = 17.2H
z), 4.78-4.97 (1H, m), 5.11 (1H, d)
d, J = 6.2, 9.2 Hz), 6.85 (2H, d, J = 8.
4Hz), 7.19 (2H, d, J = 8.4Hz), 7.30
(2H, d, J = 8.4Hz), 7.84 (2H, d, J = 8.
4Hz)

Reference Example 94 (S, S)-[3- (3- ^t -butoxycarbonylaminopropyl) -4- [2- [4-
(3-Methoxycarbonylguanidino) benzoylamino] -3- (4-methoxyphenyl) propionyl] -2
-Oxopiperazin-1-yl] acetic acid ^t- butyl ester (S, S)-[3- (3- ^t -butoxycarbonylaminopropyl) -4- [2- [4- (1,3-) obtained in Reference Example 93.
Dimethoxycarbonylguanidino) benzoylamino]
-3- (4-Methoxyphenyl) propionyl] -2-
Oxopiperazin-1-yl] acetic acid ^t- butyl ester (0.41 g, 0.50 mmol) was added to methanol (4.1 m).
1) and H ₂ O (0.41 ml), and under ice cooling,
Lithium hydroxide 1.0 hydrate (22.9mg, 0.55m
mol) was added. After stirring at 0 ° C. for 30 minutes, the reaction system was adjusted to pH 4 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure.
The residue was purified by silica gel column chromatography (ethyl acetate / methanol = 10/1) to give the title compound (0.36 g, 95%) as a colorless amorphous powder. ^{IR νmax cm - 1 (KBr)} : 3400,29
70, 1733, 1640, 1508, 1435, 1360,
1240,1150 NMR (CD ₃ OD) δ: 1.39 (9H, s), 1.46
(9H, s), 1.20-2.05 (4H, m), 2.42-
2.64 (1H, m), 2.84-4.20 (9H, m), 3.
68 (3H, s), 3.77 (3H, s), 4.80-5.0
0 (1H, m), 5.10 (1H, dd, J = 9.0,6.4H
z), 6.84 (2H, d, J = 8.8Hz), 7.18 (2
H, d, J = 8.8Hz), 7.45 (2H, d, J = 8.8H
z), 7.82 (2H, d, J = 8.8Hz)

Reference Example 95 (S, S)-[3- [3- (4
-Guanidinobenzoylamino) propyl] -4- [2-
[4- (3-Methoxycarbonylguanidino) benzoylamino] -3- (4-methoxyphenyl) propionyl] -2-oxopiperazin-1-yl] acetic acid hydrochloride Obtained in Reference Example 94 (S, S)-[ 3- (3- ^t -butoxycarbonylaminopropyl) -4- [2- [4- (3-methoxycarbonylguanidino) benzoylamino] -3-
(4-Methoxyphenyl) propionyl] -2-oxopiperazin-1-yl] acetic acid ^t- butyl ester (0.
35 g, 0.46 mmol was added to methylene chloride (2.0 m
1), and trifluoroacetic acid (2.0 ml) was added with stirring at room temperature. After 2 hours, the reaction system was concentrated under reduced pressure, and the residue was treated with 1,4-dioxane (3.5 ml) and H ₂ O (7.
0 ml). 4-Guanidinobenzoic acid N-hydroxy-5-norbornene-2,3-dicarboximide ester (0.19 g, 0.50 mm) was added to this solution.
ol) and baking soda (0.19 g, 2.28 mmol) were added at room temperature. After 1 hour, the reaction system was charged with 1N hydrochloric acid aqueous solution.
It was adjusted to H2 and concentrated under reduced pressure. The residue was subjected to column chromatography (CHP-20, H ₂ 0-5% CH ₃ CNaq-1).
0% CH ₃ CNaq-15% CH ₃ CNaq-20% CH ₃ C
Purification by _{Naq-25% CH 3 CNaq)} , as a hydrochloride salt with 1N aqueous hydrochloric acid, the title compound (0.29 g, 69
%) As a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 69.9 ° (C = 1.25%
in MeOH) Elemental analysis value: C ₃₇ H ₄₄ N ₁₀ O ₉・ 2.0HCl ・ 4.0H ₂
Calculated as O (917.801): C, 48.42; H, 5.93; N, 15.26 Analytical value: C, 48.30; H, 5.78; N, 15.20

Reference Example 96 (S)-[4-[[4- (3-
Methoxycarbonylguanidino) benzoylamino] acetyl] -3- [3- [4- (3-methoxycarbonylguanidino) benzoylamino] propyl] -2-oxopiperazin-1-yl] acetic acid hydrochloride Obtained in Reference Example 26 ( S)-[4-[(4-guanidinobenzoylamino) acetyl] -3- [3- (4-guanidinobenzoylamino) propyl] -2-oxopiperazin-1-yl] acetic acid hydrochloride (0.3 g, 0 .51 mmol)
Was added to 1,4-dioxane (3.0 ml) and H ₂ O (3.0 m).
l) and dissolved in 2N aqueous sodium hydroxide solution (2.6
0 ml, 5.10 mmol) and chlorocarbonic acid methyl ester (0.31 ml, 4.00 mmol) were gradually added under stirring at 0 ° C. while keeping the pH of the reaction system at 10 or lower. 0 ° C
After stirring for 10 minutes with, the reaction system was adjusted to pH 4 with a 1N aqueous hydrochloric acid solution, and separated and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was subjected to column chromatography (CHP-20,
10% CH ₃ CNaq-15% CH ₃ CNaq-20% CH ₃
Purification by _{CNaq-25% CH 3 CNaq-} 35% CH 3 CNaq), (S) - [4 - [[4- (1,3- dimethoxy carbonyl guanidino) benzoylamino] acetyl] -3
-[3- [4- (1,3-Dimethoxycarbonylguanidino) benzoylamino] propyl] -2-oxopiperazin-1-yl] acetic acid (0.26 g, 62%) was obtained as a colorless amorphous powder. This product (0.26g, 0.31mmol)
With methanol (2.6 ml) and H ₂ O (0.26 ml)
Lithium hydroxide 1.0 hydrate (42 mg,
1.00 mmol) was added under ice cooling. After 1 hour, the reaction system was adjusted to pH 4 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure. The residue was subjected to column chromatography (CHP-20,
5% CH ₃ CNaq-10% CH ₃ CNaq-15% CH ₃ C
Naq-20% CH ₃ CNaq) and _{(LH-20, H 2 O} )
The title compound (0.13 g, 58%) was obtained as a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 48.2 ° (C = 1.43%
in MeOH) Elemental analysis value: C ₃₁ H ₃₈ N ₁₀ O ₁₀・ 1.0HCl ・ 3.0H
Calculated as ₂ O (801.211): C, 46.47; H, 5.66; N, 17.48 Analytical value: C, 46.30; H, 5.38; N, 17.35

Reference Example 97 (S)-[3- (3- ^t -Butoxycarbonylaminopropyl) -4-[(4-guanidinobenzoylamino) acetyl] -2-oxopiperazin-1-yl] acetic acid ^t -butyl ester Reference (S)-[4-Benzyloxycarbonylaminoacetyl-3- (3- ^t -butoxycarbonylaminopropyl) -2-oxopiperazin-1-yl] acetic acid ^t- butyl ester obtained in Example 2 (0.70 g, 1. 24 mmol)
Was dissolved in ethyl acetate (7.0 ml) and 10% Pd-
C (0.21 g) was added, and the mixture was stirred under a hydrogen atmosphere at room temperature for 1 hr. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue was treated with 1,4-dioxane (7.0 ml) and H ₂ O.
Dissolve in (7.0 ml), 4-guanidinobenzoic acid
N-hydroxy-5-norbornene-2,3-dicarboximide ester (0.56 g, 1.49 mmol)
Was added at room temperature. After 1 hour, the reaction system was adjusted to pH 4 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure. The residue was subjected to column chromatography (CHP-20, H ₂ 0-5% CH ₃ CNa).
_{q-10% CH 3 CNaq-} 15% CH 3 CNaq-20% C
H ₃ CNaq) to give the title compound (0.70 g, 9
6%) as a colorless amorphous powder. IR νmax cm ^-1 (KBr): 3320,297
0,2920,1730,1640,1560,1500,1
445, 1360, 1250, 1155 NMR (CD ₃ OD) δ: 1.42 (9H, s), 1.48.
(9H, s), 1.02-2.17 (4H, m), 2.90-
3.20 (2H, m), 3.36-4.64 (6H, m), 4.
00 (1H, d, J = 17.5Hz), 4.12 (1H, d,
J = 17.5 Hz), 4.82-5.03 (1 H, m), 7.
38 (2H, d, J = 8.6Hz), 7.97 (2H, d, J
== 8.6Hz)

Reference Example 98 (S)-[3- (3- ^t -Butoxycarbonylaminopropyl) -4-[[4- (1,3
-Dimethoxycarbonylguanidino) benzoylamino] acetyl] -2-oxopiperazin-1-yl] acetic acid
^t- Butyl ester (S)-[3- (3- ^t -butoxycarbonylaminopropyl) -4-[(4-guanidinobenzoylamino) acetyl] -2-oxopiperazine-1-obtained in Reference Example 97
Ile] acetic acid ^t- butyl ester (0.70g, 1.19m
mol) in 1,4-dioxane (7.0 ml) and H ₂ O
Dissolve it in (7.0 ml) and add 2N aqueous sodium hydroxide solution (4.20 ml, 8.33 mmol) and chlorocarbonic acid methyl ester (0.46 ml, 5.94 mmol) at 0 ° C.
With stirring, the reaction system was gradually added while maintaining the pH at 10 or less. After stirring at 0 ° C for 30 minutes, the reaction system was adjusted to pH 4 with a 1N aqueous hydrochloric acid solution, and separated and extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / methanol = 13/1),
The title compound (0.67 g, 80%) was obtained as a colorless amorphous powder. IR νmax cm ^-1 (KBr): 3380,297
0,1730,1640,1490,1433,1362,1
250,1155 NMR (CD ₃ OD) δ: 1.42 (9H, s), 1.48
(9H, s), 1.30-2.15 (4H, m), 2.98-
3.20 (2H, m), 3.45 (3H, s), 3.72 (3
H, s), 3.34-4.70 (8H, m), 4.85-5.0.
5 (1H, m), 7.32 (2H, d, J = 8.5Hz), 7.
91 (2H, d, J = 8.5Hz)

Reference Example 99 (S)-[3- (3- ^t -Butoxycarbonylaminopropyl) -4-[[4- (3-methoxycarbonylguanidino) benzoylamino] acetyl] -2-oxopiperazin-1-yl ] Acetic acid ^t- butyl ester (S)-[3- (3- ^t -butoxycarbonylaminopropyl) -4-[[4- (1,3-dimethoxycarbonylguanidino) benzoylamino] acetyl] obtained in Reference Example 98
2-oxopiperazin-1-yl] acetic acid ^t- butyl ester (0.67 g, 0.95 mmol) was dissolved in methanol (6.7 ml) and H ₂ O (0.67 ml),
Under ice cooling, lithium hydroxide 1.0 hydrate (45.8 mg,
1.09 mmol) was added. After stirring at 0 ° C. for 30 minutes, the reaction system was adjusted to pH 4 with a 1N aqueous hydrochloric acid solution and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate / methanol = 10 / 1-5 / 1) to give the title compound (0.44 g, 72%) as a colorless amorphous powder. IR νmax cm ^-1 (KBr): 3390,297
0,2925,1730,1640,1525,1435,1
360, 1240, 1155 NMR (CD ₃ OD) δ: 1.42 (9H, s), 1.47.
(9H, s), 1.20-2.14 (4H, m), 2.96-
3.18 (2H, m), 3.68 (3H, s), 3.98 (1
H, d, J = 17.4 Hz), 4.12 (1 H, d, J = 17.4).
4Hz), 3.22-4.66 (6H, m), 4.82-5.
04 (1H, m), 7.45 (2H, d, J = 8.6Hz),
7.86 (2H, d, J = 8.6Hz)

Reference Example 100 (S)-[3- [3- (4-
Guanidinobenzoylamino) propyl] -4-[[4-
(3-Methoxycarbonylguanidino) benzoylamino] acetyl] -2-oxopiperazin-1-yl] acetic acid trifluoroacetic acid salt (also known as (S) -3- [3- (4
-Guanidinobenzoylamino) propyl] -4-
[[4- (3-Methoxycarbonylguanidino) benzoylamino] acetyl] -2-oxopiperazine-1-
Acetic acid trifluoroacetate) (S)-[3- (3- ^t -butoxycarbonylaminopropyl) -4-[[4- (3-methoxycarbonylguanidino) benzoylamino] acetyl] -2-obtained in Reference Example 99
Oxopiperazin-1-yl] acetic acid ^t- butyl ester (0.44 g, 0.68 mmol) was added to methylene chloride (4.4
ml) and trifluoroacetic acid (4.4 ml) was added with stirring at room temperature. After 1 hour, the reaction system was concentrated under reduced pressure, and the residue was treated with 1,4-dioxane (4.4 ml) and H ₂ O (4.4 ml).
ml). To this solution was added 4-guanidinobenzoic acid N-hydroxy-5-norbornene-2,3-dicarboximide ester (0.31 g, 0.82 mmo).
l) and baking soda (0.29 g, 3.40 mmol) were added at room temperature. After 1 hour, the reaction system was adjusted to pH with 1N hydrochloric acid aqueous solution.
It was adjusted to 2 and concentrated under reduced pressure. The residue was subjected to column chromatography (CHP-20, H ₂ 0-5% CH ₃ CNaq-1).
0% CH ₃ CNaq-15% CH ₃ CNaq) and (LH-2
0, H ₂ O) to give the title compound (0.18 g, 32
%) As a colorless amorphous powder. Specific rotation: [α] _D ²⁰ + 46.9 ° (C = 0.976%
in MeOH) Elemental analysis value: C ₂₉ H ₃₆ N ₁₀ O ₈ 1.0CF ₃ CO ₂ H
Calculated as 3.0H ₂ O (820.737): C, 45.37; H, 5.28; N, 17.07 Analytical value: C, 45.42; H, 5.08; N, 16. 92

[0182]

INDUSTRIAL APPLICABILITY In the present invention, a lyophilized amorphous water-soluble 2-piperazinone-1-acetic acid derivative is dispersed and atomized in a polymer solution to obtain a sustained release with a high content and a small initial release. Microcapsules can be obtained. Furthermore, by using this microcapsule, for example, antithrombotic drug, thrombosis, angina, unstable angina or ischemic complication after percutaneous intracoronary angioplasty or coronary thrombolytic therapy Alternatively, side effects such as long-term bleeding caused by a large amount of the initial release of the above compound useful as a prophylactic / therapeutic agent for reocclusion or restenosis can be reduced.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A61K 31/495 ACV A61K 31/495 ACV 9/52 9/52 A 47/16 47/16 C 47 / 32 47/32 D B01J 13/04 C07D 241/08 C07D 241/08 413/12 241 413/12 241 413/14 241 413/14 241 B01J 13/02 A // (C07D 413/12 241: 08 271 : 06) (C07D 413/14 241: 08 271: 06)

Claims (35)

[Claims] 1. Amorphous water-soluble formula (I): [In the formula, A ¹ and A ² are each a proton accepting group or a group capable of converting into a proton accepting group, and D is a hetero atom and / or an atomic chain of 2 to 6 optionally having a 5- or 6-membered ring. Spacer (however, the 5 or 6 membered ring is 2 depending on the bonding position)
Or -CH 3 converted with atomic chains), the R ¹ is a hydrogen atom or a hydrocarbon group, R ² represents a hydrogen atom or an α- amino acid
Represents a residue excluding (NH ₂ ) COOH, or R ¹ and R ²
May be bonded to form a 5- or 6-membered ring, and P is a spacer of 1 to 10 atom chain which may be through a heteroatom and / or a 5- or 6-membered ring (provided that the 5- or 6-membered ring is Represents a 2 or 3 atom chain depending on the bonding position), Y represents a carboxyl group which may be esterified or amidated, and n represents an integer of 0 to 8. ] A microcapsule comprising a compound or a salt thereof, a 2-piperazinone-1-acetic acid derivative, and a polymer. 2. The microcapsule according to claim 1, which is sustained-release. 3. The microcapsule according to claim 1, wherein the 2-piperazinone-1-acetic acid derivative is dispersed in a polymer. 4. The microcapsule according to claim 1, wherein the 2-piperazinone-1-acetic acid derivative is readily soluble in water. 5. The microcapsule according to claim 1, wherein the solubility of the 2-piperazinone-1-acetic acid derivative in water is about 1 g / 100 ml or more at 20 ° C. 6. The microcapsule according to claim 1, wherein the 2-piperazinone-1-acetic acid derivative has an average particle size of about 30 μm or less. 7. The microcapsule according to claim 1, wherein the 2-piperazinone-1-acetic acid derivative has an average particle size of about 5 μm or less. 8. A 2-piperazinone-1-acetic acid derivative,
The microcapsule according to claim 1, which is (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid. 9. A 2-piperazinone-1-acetic acid derivative,
The microcapsule according to claim 1, which is (S) -4- (4-guanidinobenzoylamino) acetyl-3- [3- (4-guanidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid hydrochloride. 10. A 2-piperazinone-1-acetic acid derivative,
(S) -4- (4-amidinobenzoyl) aminoacetyl-3- {3- (4-amidinobenzoyl) amino} propyl-2-oxopiperazine-1-acetic acid.
The described microcapsules. 11. A 2-piperazinone-1-acetic acid derivative,
(S) -4- (4-amidinobenzoyl) aminoacetyl-3- {3- (4-amidinobenzoyl) amino} propyl-2-oxopiperazine-1-acetic acid trifluoroacetic acid salt according to claim 1. capsule. 12. A 2-piperazinone-1-acetic acid derivative,
(S) -4- [4- (2-Aminoethyl) benzoylamino] acetyl-3- [3- (4-amidinobenzoylamino)] propyl-2-oxopiperazine-1-acetic acid according to claim 1. Micro capsule. 13. A 2-piperazinone-1-acetic acid derivative,
The microcapsule according to claim 1, which is (S) -4- (4-amidinobenzoylamino) acetyl-3- [2- (4-guanidinobenzoylamino)] ethyl-2-oxopiperazine-1-acetic acid. 14. A 2-piperazinone-1-acetic acid derivative,
(S) -4- (4-amidinobenzoyl) aminoacetyl-3- [3- (4-guanidinobutanoylamino)] propyl-2-oxopiperazine-1-acetic acid.
The described microcapsules. 15. The microcapsule according to claim 1, wherein the polymer is a biodegradable polymer. 16. The microcapsule according to claim 15, wherein the biodegradable polymer is a polyester. 17. The microcapsule according to claim 16, wherein the polyester is a lactic acid / glycolic acid polymer. 18. A lactic acid / glycolic acid molar ratio of about 100.
18. The microcapsule according to claim 17, which is / 0 to about 25/75. 19. The microcapsule according to claim 17, wherein the lactic acid / glycolic acid polymer has a weight average molecular weight of about 5,000 to about 30,000. 20. The microcapsule according to claim 16, wherein the polyester is a polymer of hydroxybutyric acid / glycolic acid. 21. The microcapsule according to claim 20, wherein the molar ratio of hydroxybutyric acid / glycolic acid is about 100/0 to about 25/75. 22. The microcapsule according to claim 20, wherein the hydroxybutyric acid / glycolic acid polymer has a weight average molecular weight of about 5,000 to about 25,000. 23. The microcapsule according to claim 1, which is used for preventing / treating cardiovascular diseases. 24. Prevention of ischemic complication or reocclusion or restenosis of coronary artery after thrombosis, angina, unstable angina or percutaneous intracoronary angioplasty or after coronary thrombolytic therapy. The microcapsule according to claim 1, which is for treatment. 25. A dispersion obtained by dispersing an amorphous water-soluble compound represented by formula (I) or a salt thereof, a 2-piperazinone-1-acetic acid derivative, in an organic solvent solution of a high molecular weight polymer is used. Microcapsules obtained by dispersing in a phase to form an s / o / w type emulsion and drying in water. 26. The microcapsule according to claim 25, wherein the concentration of the 2-piperazinone-1-acetic acid derivative in the organic solvent solution of the polymer is about 0.01 to about 70% (w / w). 27. The microcapsule according to claim 25, wherein the solution of the polymer in the organic solvent further contains a basic substance. 28. The microcapsule according to claim 27, wherein the basic substance is a basic amino acid. 29. The microcapsule according to claim 27, wherein the basic substance is L-arginine, L-lysine or N-methylglucamine. 30. The microcapsule according to claim 25, wherein the concentration of the basic substance in the organic solvent solution of the polymer is about 0.1 to about 3% (w / w). 31. The microcapsule according to claim 25, wherein the aqueous phase further contains an osmotic pressure adjusting agent. 32. The microcapsule according to claim 31, wherein the osmotic pressure adjusting agent is sodium chloride. 33. A dispersion obtained by dispersing an amorphous water-soluble compound represented by the formula (I) or a salt thereof, a 2-piperazinone-1-acetic acid derivative, in an organic solvent solution of a polymer, A method for producing microcapsules, which comprises dispersing in a phase to form an s / o / w type emulsion and drying in water. 34. Use of the amorphous water-soluble compound represented by formula (I) or a salt thereof, a 2-piperazinone-1-acetic acid derivative, for the production of microcapsules according to claim 1. 35. The ischemic complication or coronary revascularization after thrombosis, angina, unstable angina or percutaneous intracoronary angioplasty or after coronary thrombolytic therapy of the microcapsule according to claim 1. Use for the manufacture of a medicament for preventing or treating obstruction or restenosis.