JPH08291059A - Peroral therapeutic agent containing 2,3-diaminopropionic acid derivative - Google Patents

Abstract

PURPOSE: To obtain the subject therapeutic agent, containing a specific compound, capable of manifesting excellent absorbability, more selectively interacting with a receptor of a cell adhesion active protein such as a fibrinogen and useful as a suppressant, etc., for blood platelet aggregation more improved in stability. CONSTITUTION: This therapeutic agent contain a compound of formula I (R<1> is H or an ester; R<2> is a lower alkoxy-substituted phenyl, 1-naphthyl or 2- naphthyl; A is CONH, OCH2 , OCH2 CH2 or CH2 CH2 ) [e.g. (2S)-3-(3-(4- amidinobenzoylamino)-propanoylaminio)-2-(3,4-dimethoxy) benzenesulfonylaminopropanoic acid trifluoroacetate] or its pharmaceutically permissible salt. Furthermore, the compound is preferably obtained by condensing, e.g. a compound of formula II with a compound of formula III using reaction for forming amide bond and then, as necessary, removing the protecting group or introducing an ester group thereinto.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an oral therapeutic agent containing a 2,3-diaminopropionic acid derivative useful as a platelet aggregation inhibitor, cancer metastasis inhibitor, wound healing agent or bone resorption inhibitor.

[0002]

BACKGROUND OF THE INVENTION Involved in adhesion between cells and stromal connective tissue,
Proteins having various physiological activities related to cell functions of animal cells are called cell adhesion activating proteins, and for example, fibronectin, vitronectin, laminin, etc. are known. The core sequences of their cell binding sites are arginine-glycine-aspartic acid [Arg-Gly-Asp.
(Hereinafter sometimes abbreviated as RGD)]] (Pierschbachr, MD et al .: Nature, 309, 30 (1
984), Suzuki, S. et al .: J. Biol. Chem., 259, 15307 (1984
), Plow, E. et al .: Proc.Natl.Acad.Sci. USA, 82 , 8057.
(1985)). It has been known that this RGD interacts with the receptor of cell adhesion protein and exhibits various pharmacological actions. For example, fibrinogen present in plasma is mediated by RGD via platelet membrane glycoprotein complex IIb /
Interacts with IIIa to cause platelet aggregation,
It is considered that a synthetic peptide having RGD inhibits the interaction between fibrinogen and platelet membrane glycoprotein complex IIb / IIIa and is effective as a platelet aggregation inhibitor. (Phillips, DR: Cell, 65 , 359 (1991)
). It is also known that peptides having RGD derived from snake venom significantly inhibit bone resorption by osteoclasts (Sato, M. et al .: J. Cell Biol. , 111 , 1713 (1990)).

Fibronectin is considered to be involved in cell differentiation and proliferation (Yamada, KM: A).
Rev. Biochem., 52, 761 (1983)) stimulates the migration ability of fibroblasts and macrophages, and is expected to be applied to wound healing and regulation of immune function. In particular, local treatment of corneal disorders using the wound healing promoting effect has already been tried (Fujikawa, LS et al .: Lab. In
vest., 45, 120 (1981)). Furthermore, cell adhesion proteins have been attracting attention as substances related to cancer metastasis. It is known that cancer cells form a multicellular mass in the presence of fibronectin and laminin, which makes proliferation and survival easier. In fact, the RGD sequence, which is the adhesion core of fibronectin, suppresses metastasis of cancer cells. It has been confirmed that (H
umphries, MJ et al .: Science , 233 , 467 (1986)). Thus, cell adhesion proteins have various physiological activities, and drugs that selectively interact with their receptors are expected to be applied to preventive or therapeutic agents for various diseases.

On the other hand, compounds acting on these receptors include EP 445796, EP 512831, EP 540334 and WO 94.
Although / 8962 and WO 94/12181 have been reported, none has been reported as an oral therapeutic agent sufficient for clinical application.

[0005]

The object of the present invention is an oral preparation exhibiting excellent absorbability, which interacts more selectively with the receptors for cell adhesion-activating proteins such as fibrinogen and fibronectin, and is more stable. It is intended to provide an oral therapeutic agent containing a 2,3-diaminopropionic acid derivative which is useful as a platelet aggregation inhibitor, a cancer metastasis inhibitor, a wound healing agent or a bone resorption inhibitor having excellent properties.

[0006]

[Means for Solving the Problems] As a result of intensive studies, the present inventors have found that it is an oral preparation exhibiting excellent absorbability, and more selectively interacts with receptors for cell adhesion-activating proteins such as fibrinogen and fibronectin. Then, they have found an orally available therapeutic agent containing a 2,3-diaminopropionic acid derivative which is more stable and is useful as a platelet aggregation inhibitor, cancer metastasis inhibitor, wound healing agent or bone resorption inhibitor.

That is, the gist of the present invention resides in the formula (1)

[Chemical 6] [In the formula, R ¹ represents a hydrogen atom or an ester group, R ² represents a phenyl group substituted with 1 to 3 same or different lower alkoxy groups, a 1-naphthyl group or a 2-naphthyl group, and A is A _{-CONH -, - OCH 2 -,} - OCH
₂ CH ₂ - or -CH ₂ CH ₂ - represents a. ] An oral therapeutic agent containing a compound represented by the formula or a pharmaceutically acceptable salt thereof, an oral therapeutic agent according to Formula (1), wherein R ¹ is a hydrogen atom, Formula (2),

[Chemical 7] Formula (3),

Embedded image Formula (4),

[Chemical 9] Formula (5) or

[Chemical 10] The present invention relates to an oral therapeutic agent or an oral therapeutic agent, which comprises the compound represented by or a pharmaceutically acceptable salt thereof or an oral platelet aggregation inhibitor.

Specific examples of the ester group include lower alkyl groups such as methyl group, ethyl group and propyl group, cyclohexyl, benzyl, acetoxymethyl, 1-acetoxyethyl, 1-acetoxy-1-phenylmethyl, pivaloyl. A lower alkyl group substituted with -OCOP ¹ (wherein P ¹ represents a lower alkyl group, a cycloalkyl group, an aryl group, or a heterocyclic group) such as oxymethyl and 1-pivaloyloxyethyl; Lower substituted with -OCOOP ² (in the formula, P ² represents a lower alkyl group, a cycloalkyl group, an aryl group, or a heterocyclic group) such as 1-methoxycarbonyloxyethyl and 1-ethoxycarbonyloxyethyl. Alkyl group, 5-oxo-2-tetrahydrofuranyl, 1,3-dihydro-3-
Heterocyclic group such as oxo-1-isobenzofuranyl group, 5-
Methyl-2-oxo-1,3-dioxolen-4-ylmethyl, 2- (1-morpholinyl) ethyl, 5-indanyl, 3-dimethylamino-2- (dimethylaminomethyl) propyl, 2-dimethylaminoethyl, 2 -Diethylaminoethyl, methoxycarbonylmethyl, 2-dimethylaminocyclohexyl and the like can be mentioned.

Examples of the lower alkyl include a linear or branched alkyl group having 1 to 6 carbon atoms, specifically, methyl, ethyl, propyl, 1-methylethyl,
Butyl, 1-methylpropyl, 2-methylpropyl,
1,1-dimethylethyl, pentyl, 1-methylbutyl, 1-ethylpropyl, hexyl, 1-methylpentyl, 1-ethylbutyl, 2-methylpentyl and the like can be mentioned. Examples of the cycloalkyl group include cycloalkyl groups having 5 to 7 carbon atoms, and specific examples thereof include cyclopentyl, cyclohexyl, cycloheptyl and the like. Examples of the aryl group include aryl groups having 6 to 14 carbon atoms, and specific examples thereof include phenyl, 1-naphthyl, 2-naphthyl, 1-anthranyl, 2-anthranyl and the like.

The heterocyclic group is, for example, a 5- to 7-membered 1-ring, 9- or 10-membered bicyclic ring containing 1 to 4 heteroatoms arbitrarily selected from the group of oxygen atom, nitrogen atom and sulfur atom. Examples thereof include a saturated or unsaturated heterocyclic group of the formula or a 12 to 14-membered tricyclic group. The nitrogen atom and the sulfur atom may be optionally oxidized, and the bonding point is on any nitrogen atom or carbon atom. Further, ring-constituting carbon atoms may be substituted with an oxo group. Specifically, a 6-membered 1-ring saturated heterocyclic group containing 1 or 2 heteroatoms arbitrarily selected from the group of oxygen atom, nitrogen atom, sulfur atom such as piperidyl, piperazinyl, morpholinyl, pyridyl, pyridazinyl, Oxygen atom such as pyrazinyl, nitrogen atom, 6-membered 1-ring unsaturated heterocyclic group containing 1 or 2 heteroatoms arbitrarily selected from the group of sulfur atom, oxygen atom such as oxolanyl, pyrrolidinyl, nitrogen atom, 5-membered 1-ring saturated heterocyclic group containing 1 or 2 heteroatoms arbitrarily selected from the group of sulfur atoms, imidazolyl, furyl, 2-oxo-1,
5 containing 1 to 3 heteroatoms arbitrarily selected from the group of oxygen atom, nitrogen atom, and sulfur atom such as 3-dioxorenyl, pyrrolyl, 5-oxo-2-tetrahydrofuranyl, etc.
Membered monocyclic unsaturated heterocyclic group, quinolyl, isoquinolyl, indolyl, 1,3-dihydro-3-oxo-1-isobenzofuranyl, etc., selected from the group of oxygen atom, nitrogen atom and sulfur atom. A 9- or 10-membered bicyclic unsaturated heterocyclic group containing 1 to 3 heteroatoms, 1 selected arbitrarily from the group of oxygen atoms such as anthraquinolyl, nitrogen atoms, and sulfur atoms
To 12 to 14-membered tricyclic unsaturated heterocyclic group containing 3 to 3 heteroatoms.

As the halogen atom, for example, a fluorine atom,
Examples thereof include chlorine atom, bromine atom and iodine atom. When the compound represented by the formula (1) has an asymmetric carbon atom,
Any isomer or mixture thereof may be used, and when two or more asymmetric carbons are present, as an enantiomer, diastereomer, or a mixture thereof,
For example, it may exist as a racemate. The preferred configuration of the asymmetric carbon at the 2-position of 2,3-diaminopropionic acid is S.

The compound of the present invention represented by the formula (1) can be produced from easily available raw materials and reagents by a known synthesis reaction, for example, by the method described below or a modified method thereof. it can. The following will be described in detail in the following cases.

A method for producing a compound (formula (6)) in which A represents —CONH— in formula (1). The compound of formula (6) can be produced by the following steps.

[Chemical 11] [In the formula, R ¹ and R ² have the same meanings as described above, R ³ represents a carboxyl group protecting group, and X ¹ represents a hydrogen atom or an amidino group protecting group. That is, this step is performed by condensing the compound of formula (7) and the compound of formula (8) using an amide bond-forming reaction, and then deprotecting the protecting group or introducing an ester group as necessary. can do.

The amide bond-forming reaction can be carried out by a known method, for example, "basics and experiments of peptide synthesis".
(Izumiya et al., Maruzen, 1985). For example, an amino group-free compound and a carboxyl group-free compound are treated with 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride or the like in an inert solvent such as N, N-dimethylformamide, dichloromethane or acetonitrile. 0 to 40 ° C using a condensing agent
In, it can be carried out by stirring for 1 to 24 hours. If necessary, the reaction can be carried out in the presence of an additive such as 1-hydroxybenzotriazole or in the presence of a base such as triethylamine.

The compound of formula (7) can be produced, for example, by the method described in Example 1. The compound of the formula (8) includes, for example, the following steps.

[Chemical 12] [In the formula, R ² and R ³ have the same meanings as described above, X represents a chlorine atom, a bromine atom or an iodine atom, and X ² and X ²
Represents different amino-protecting groups. That is, asparagine derivative (formula (9)) available as a commercial product is synthesized from Synthesis, 1981 , 26.
Using the method described in 6 or the like, or using commercially available 2,3-diaminopropionic acid and its derivative, protection or deprotection of the protecting group is performed as necessary to obtain the compound of formula (10) To produce a compound of Then, the formula (1
An amide bond-forming reaction is carried out with the compound of 1) and, if necessary, the protective group is deprotected to produce a compound of formula (12). Further, the compound of formula (8) can be produced by reacting with the sulfonic acid halide of formula (13) to deprotect the protecting group of X ² . Here, the reaction with the sulfonic acid halide can be performed, for example, with N,
It can be carried out by stirring at 0 to 80 ° C. for 1 to 24 hours in an inert solvent such as N-dimethylformamide, dichloromethane, ethyl acetate, 1,4-dioxane or a mixed solvent thereof. If necessary, a base such as triethylamine, pyridine, sodium hydrogencarbonate, etc. may be used.

A method for producing a compound (formula (14)) in which A represents —OCH ₂ — or —OCH ₂ CH ₂ — in formula (1). The compound of formula (14) can be produced by the following steps.

[Chemical 13] ^{Wherein, R 1, R 2, R} 3 and X ¹ are as defined above, n represents represents 3 or 4. That is, this step is performed by condensing the compound of formula (15) and the compound of formula (16) using an amide bond-forming reaction, and then deprotecting the protecting group or introducing an ester group as necessary. can do.

The compound of formula (15) is protected or protected by converting the nitrile group into an amidino group by reacting the p-cyanophenol represented by formula (17) with the compound of formula (18) according to the following steps. It can be produced by deprotecting the group.

Embedded image [In the formula, X, X ¹ , R ⁴ and n are as defined above. ] Here, conversion of a nitrile group to an amidino group is performed by the formula (7)
It can be carried out in the same manner as in the production of the compound of The compound of formula (16) can be produced by the following steps.

[Chemical 15] [In the formula, R ² , R ³ , X ² and X ³ are as defined above. A compound of formula (19) is obtained by converting the compound of formula (9) into a 2,3-diaminopropionic acid derivative in the same manner as in the production of the compound of formula (10) and protecting or deprotecting the protecting group. And then reacting with the sulfonic acid halide of formula (13) to deprotect the protecting group represented by X ² to produce the compound of formula (16).

In the formula (1), A is —CH ₂ CH ₂
A method for producing a compound represented by formula (Formula (20)). Formula (20)
The compound can be produced by the following steps.

Embedded image [In the formula, R ¹ , R ² , R ³ and X ¹ have the same meanings as described above. That is, this step is performed by condensing the compound of formula (21) and the compound of formula (16) using an amide bond-forming reaction, and then deprotecting the protecting group or introducing an ester group as necessary. can do.

The compound of the formula (21) can be produced by converting the nitrile group of the compound of the formula (22) into an amidino group and protecting it as necessary according to the following steps.

[Chemical 17] [In the formula, X has the same meaning as described above. ] Here, conversion of a nitrile group to an amidino group is performed by the formula (7)
It can be carried out in the same manner as in the production of the compound of Further, the compound of formula (7) is described in J. Med. Chem. , 36 , 1811 (1
993).

The above description is merely one example, and the reaction can be carried out by changing the order of each reaction, or can be produced by other methods. Examples of the protecting group for the amidino group, the protecting group for the amino group, and the protecting group for the carboxyl group include, for example, “Basics and Experiments of Peptide Synthesis” by Izumiya et al. (Maruzen, 1985) or Green's “Protective Groups in Organic Synthesis”. (Joan Willey & Sons, 1991)
The amino acid side chain protecting group described in 1) and the like can be used as the protecting group.

Examples of the amidino group-protecting group include urethane-type protecting groups such as t-butoxycarbonyl and benzyloxycarbonyl. As the amino-protecting group,
Examples of the urethane-type protecting group or acyl-type protecting group,
Examples of the urethane-type protecting group include benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-methanesulfonylethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, t-butyloxycarbonyl, and fullerene. Examples of the acyl-type protecting group include formyl, acetyl, benzoyl, trifluoroacetyl and the like. Examples of the carboxyl-protecting group include ester-type protecting groups, for example,
Lower alkyl, 2,2,2-trichloroethyl, benzyl, 4-methoxybenzyl, diphenylethyl and the like can be mentioned.

The method of introducing each protecting group and the method of deprotecting the protecting group are described in "Protective Groups in Organic Synthesis" by Green et al.
Willey & Sons, 1991) and Izumiya et al., "Basics and Experiments of Peptide Synthesis" (Maruzen, 1985). For the method for introducing a protecting group for an amidino group and the method for deprotecting a protecting group, see
The butoxycarbonyl group is introduced by the same method as in the case of the amino group, or the free amidino group is introduced with N, O in an inert solvent such as dichloromethane or acetonitrile.
-Bistrimethylsilylacetamide can be added and stirred for 0.5 to 3 hours, and then di-t-butyldicarbonate can be added and stirred at 0 to 40 ° C for 0.5 to 24 hours. The protection can be carried out by stirring with an acid such as TFA or 4N HCl-dioxane at 0 to 40 ° C. for 0.5 to 6 hours.

Regarding the method of introducing a protecting group for an amino group and the method of deprotecting a protecting group, for example, a t-butoxycarbonyl group can be converted into a free amino group by di-t in an inert solvent such as 1,4-dioxane-water. -Butyldicarbonate and can be introduced by stirring at 0 to 40 ° C. for 0.5 to 24 hours, and for deprotection, T
It can be carried out by stirring with an acid such as FA or 4N HCl-dioxane at 0 to 40 ° C. for 0.5 to 6 hours. In addition, the benzyloxycarbonyl group is introduced by adding carbobenzoxylchloride to the free amino group in an inert solvent such as 1,4-dioxane or methanol and stirring the mixture at 0 to 40 ° C. for 0.5 to 24 hours. Deprotection can be carried out by hydrogenation in the presence of a noble metal catalyst such as palladium on carbon (Pd / C). The method for introducing a protecting group for a carboxyl group and the method for deprotecting a protecting group include, for example, methyl and ethyl esters having a free carboxyl group as an inert solvent such as dichloromethane, methanol or ethanol, dimethylaminopyridine, and 1- (3- It can be introduced by adding a condensing agent such as dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and stirring at 0 to 40 ° C. for 0.5 to 24 hours. For deprotection, an inert solvent such as methanol is used. 0 with a base such as 1N aqueous sodium hydroxide solution
It can be carried out by stirring at 0.5 to 40 ° C. for 0.5 to 6 hours.

The protective group is selected so that other protective groups are not eliminated during the reaction and deprotection. The compound represented by the formula (1) can be purified by a conventional purification method such as recrystallization or high performance liquid chromatography.

Examples of the pharmaceutically acceptable salt include pharmaceutically acceptable acid addition salts or base addition salts. Examples of the acid addition salt include inorganic acid addition salts such as hydrochloride, sulfate, and phosphate, or acetate, butyrate, methanesulfonate, trifluoroacetate, citrate, fumarate, maleate, Examples thereof include organic acid addition salts such as succinate and salicylate. Examples of the base addition salt include inorganic base addition salts or organic base addition salts, and as the inorganic base addition salts, sodium salts, alkali metal salts such as potassium salts, magnesium salts, alkaline earth metal salts such as calcium salts,
Examples thereof include ammonium salts, and examples of the organic base addition salt include basic amino acid addition salts such as arginine salt and lysine salt. Such salts can be easily produced by known means. For example, in the case of acetate salt, it is produced by dissolving the compound of formula (1) in water and adding the required amount of acetic acid.

The animals to which the oral therapeutic agent of the present invention is administered are not particularly limited, and not only humans but also mice, rats, dogs, cats, cows, horses, goats, sheep, rabbits,
Various mammals such as pigs are targeted. The dose and the number of doses vary depending on the animal species, administration route, degree of symptoms, body weight, etc. and are not particularly limited.
About 1 mg to 1 g per day is administered once or more times a day. As the dosage form, for example, powder,
Fine granules, granules, tablets, capsules and the like can be mentioned. In the case of formulation, it is produced by a conventional method using a usual formulation carrier. That is, if necessary, a binder, a disintegrant, a lubricant,
After adding a coloring agent and the like, tablets, granules, powders, capsules and the like are prepared by a conventional method.

[0027]

Industrial Applicability According to the present invention, it is possible to provide an oral therapeutic agent containing a 2,3-diaminopropionic acid derivative which is useful as a platelet aggregation inhibitor, cancer metastasis inhibitor, wound healing agent or bone resorption inhibitor. became.

[0028]

The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. The abbreviations have the following meanings. Abbreviations Name DMF: N, N-dimethylformamide HOBT · H ₂ O: 1- hydroxybenzotriazole hydrate WSC · HCl: 1- (3- dimethylaminopropyl) -3-Echirukaru Bojiimido hydrochloride TFA: Trifluoroacetic acid Z : benzyloxycarbonyl Boc: ₂ t-butoxycarbonyl (Boc) O: di -t- butyl dicarbonate THF: tetrahydrofuran

Example 1 (2S) -3- (3- (4-amidinobenzoylami)
No) propanoylamino) -2- (3,4-dimethoxy)
Si) Benzenesulfonylaminopropanoic acid TFA salt
Adult (2S)-3-amino-2-benzyloxy carbonylation
Ruaminopropanoic acid NZ-asparagine (3.124 g) was added to DMF (30
ml) -water (30 ml), bis (trifluoroacetyl) iodobenzene (7.575 g) was added at room temperature, and the mixture was stirred for 15 minutes. Then at room temperature pyridine (1.8
74 ml) was added and the mixture was stirred for 4 hours. The solvent was evaporated under reduced pressure, water (150 ml) was added to the residue, and the mixture was washed with ether (2).
Washed twice. The aqueous layer was concentrated under reduced pressure, and the residue was crystallized in ethanol-ether to obtain 2.149 g of the desired product. (2S) -2-benzyloxycarbonylamino-3
A compound of- (t-butoxycarbonylamino) propanoic acid (0.61 g) was added to 1,4-dioxane (6 m).
l) -dissolve in water (2 ml) and at room temperature (Boc) ₂ O
(0.62 g) was added and the mixture was stirred for 1.5 hours. After adding a 1N aqueous sodium hydroxide solution (4 ml), the mixture was washed twice with ether. The aqueous layer was adjusted to pH 2 with 1N hydrochloric acid and then extracted 3 times with ethyl acetate. The organic layer was washed twice with saturated saline and dried over anhydrous magnesium sulfate. The desiccant was filtered off and then concentrated under reduced pressure to obtain 0.867 g of the desired product.

(2S) -2-benzyloxycarboni
Luamino-3- (t-butoxycarbonylamino) pro
The compound (0.867 g) obtained with panic acid ethyl ester was dissolved in dichloromethane (15 ml), and ethanol (1.5 m) was added under ice cooling.
l), 4-dimethylaminopyridine (34 mg), WS
C.HCl (0.633 g) was added, and the mixture was stirred under ice cooling for 2 hours and then at room temperature for 12 hours. The solvent was concentrated under reduced pressure to about 5 ml, the residue was poured into water, and the mixture was extracted 3 times with ethyl acetate. The organic layer was washed twice with 1N HCl, twice with a saturated aqueous sodium hydrogen carbonate solution, twice with a saturated saline solution, and dried over anhydrous magnesium sulfate. The desiccant was filtered off and then concentrated under reduced pressure to obtain 0.875 g of the desired product. (2S) -2-benzyloxycarbonylamino-3
-(3- (t-butoxycarbonylamino) propanoi
(Amino) propanoic acid ethyl ester and the compound (6.0 g) obtained by acetonitrile (1
After dissolving in 0 ml), a solution of methanesulfonic acid (8.181 g) in acetonitrile (15 ml) was added dropwise at 20 ° C or lower, and the mixture was stirred at room temperature for 30 minutes. DMF below 20 ℃
(20 ml) and triethylamine (8.608 g) were sequentially added dropwise, and the mixture was stirred for 10 minutes. N-Boc-β-alanine (3.547 g), HOBT · H ₂ O (3.13)
g), and then add WSC / HCl at 5-10 ° C.
(3.928 g) was added and the mixture was stirred for 30 minutes. After further stirring at room temperature for 12 hours, it was poured into water and extracted three times with ethyl acetate. The organic layer was washed twice with 1N HCl, twice with a saturated aqueous sodium hydrogen carbonate solution, twice with a saturated saline solution, and dried over anhydrous magnesium sulfate. The desiccant was filtered off and then concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (silica gel 400 g, solvent: chloroform-acetone =).
Purified 3: 1 to 2: 1). The fraction containing the target substance was concentrated under reduced pressure to obtain 6.974 g of the target substance.

(2S) -2- (3,4-dimethoxy)
Benzenesulfonylamino-3- (3- (t-butoxy
Carbonylamino) propanoylamino) propanoic acid
The compound (702 mg) obtained as a tyl ester was added to ethanol (26 mg).
(10 ml) (50 ml of water) (350 mg), and the mixture was stirred at room temperature for 3 hours under a hydrogen gas atmosphere. The insolubles were removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was dissolved in dichloromethane (10 ml), triethylamine (0.5 ml) and (3,4-dimethoxy) benzenesulfonyl chloride (639 mg) were added at room temperature, and the mixture was stirred for 1 hr. N- (3-aminopropyl) morpholine (0.13 ml) was added, and the mixture was further stirred for 30 minutes.
The mixture was diluted with ethyl acetate, washed successively with 10% aqueous citric acid solution, saturated aqueous sodium hydrogen carbonate solution and saturated brine, and dried over anhydrous sodium sulfate. After separating the desiccant by filtration, it was concentrated under reduced pressure,
The target product (778 mg) was obtained. ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.15
(3H, t, J = 7Hz), 1.46 (9H, s),
2.40 (2H, t, J = 6Hz), 3.30-3.7
0 (4H, m), 3.93 (3H, s), 3.94 (3
H, s), 3.95-4.05 (1H, m), 4.03.
(2H, q, J = 7Hz), 5.26 (1H, m),
5.76 (1H, bd, J = 8Hz), 6.37 (1
H, m), 6.92 (1H, d, J = 9 Hz), 7.2
9 (1H, d, J = 2Hz), 7.46 (1H, dd,
J = 2.9 Hz).

4-Amidinobenzoic Acid Hydrochloride 4-Amidinobenzamide (commercial product: 7.8 g) was dissolved in water (200 ml), concentrated hydrochloric acid (200 ml) was added, and the mixture was stirred at 100 ° C. for 9 hours. The mixture was cooled and the precipitated crystals were collected by filtration. Yield 7.6g. Mass spectrum (SIMS): 165 [M + 1] ^{+ 1} H-NMR (CD ₃ OD) δ (ppm): 7.94
(2H, d, J = 8.6 Hz), 8.26 (2H, d,
J = 8.6 Hz). The compound (1.772 g) obtained with Nt-butoxycarbonyl-4-amidinobenzoic acid was suspended in dichloromethane (35 ml), N, O-bistrimethylsilylacetamide (7.2 g)) was added, and the mixture was stirred at room temperature for 1 hour. Stir for hours. Then, (Boc) ₂ O (3.9 g) was added at room temperature, and the mixture was stirred for 24 hours. The insolubles were removed by filtration, and the filtrate was concentrated under reduced pressure. Diethyl ether was added to the residue and the mixture was stirred, and the resulting crystals were collected by filtration. Yield 2.334g. ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 1.5
5, 1.75 (9H, s), 7.88 (2H, m),
8.09 (2H, m), 6.11-6.77, 7.22
-7.40 (1H, m), 9.40, 9.58, 10.
48-10.68 (1H, m).

(2S) -2- (3,4-dimethoxy)
Benzenesulfonylamino-3- (3- (4- (Nt
-Butoxycarbonylamidino) benzoylamino) p
After dissolving the compound (469 mg) obtained from the ropanoylamino) propanoic acid TFA salt in acetonitrile (5 ml), a solution of methanesulfonic acid (446 mg) in acetonitrile (2.5 ml) was added dropwise at 20 ° C or lower, and the mixture was stirred at room temperature for 30 minutes. It was stirred. DM below 20 ℃
F (6.5 ml), triethylamine (0.67 ml)
Was added dropwise and stirred for 10 minutes. After adding the compound (262 mg) obtained in 1. and HOBT · H ₂ O (170 mg), WSC · HCl (213 m
g) was added and the mixture was stirred for 30 minutes, and further stirred at room temperature for 20 hours. The reaction mixture was concentrated under reduced pressure, and the residue was TFA under ice cooling.
It was dissolved in (25 ml), stirred for 30 minutes, and further stirred at room temperature for 1 hour. After TFA was distilled off, acetic acid (15 ml) -1N hydrochloric acid (15 ml) was added to the residue, and the mixture was heated at 60 ° C for 21 hours. The reaction mixture was concentrated under reduced pressure and the residue was purified by HPLC to give a white powder. Yield 200
mg. HPLC retention time: 18.1 minutes (column: YMC-ODS 4.6 mmΦ × 250 m)
m, detection: UV 220 nm, eluent: solution A 0.1% T
FA / water, B solution 0.1% TFA / acetonitrile, flow rate: 1 ml / min. , Gradient: B solution concentration 10
% To 1% per minute. ) ¹ H-NMR (CDCl ₃ ) δ (ppm): 2.31
(2H, m), 3.00-3.60 (4H, m), 3.
80 (3H, s), 3.81 (3H, s), 3.75-
3.95 (1H, m), 7.08 (1H, d, J = 8H
z), 7.27 (1H, d, J = 2 Hz), 7.33
(1H, dd, J = 2, 8Hz), 7.88 (2H.
d, J = 8 Hz), 8.01 (2H, d, J = 8H
z), 7.95-8.15 (2H, m), 8.72 (1
H, bt, J = 5.6 Hz), 9.23 (2H, b
s), 9.40 (2H, bs).

Example 2 (2S) -3- (4- (4-amidinophenoxy) butane
Noylamino) -2- (3,4-dimethoxy) benzene
Synthesis of sulfonylaminopropanoic acid TFA salt 4- (4-cyanophenoxy) butanoic acid ethyl ester
Le 4-cyanophenol (5.0 g) and DMF (6 ml)
Dissolved in ethyl 4-bromobutanoate (7.94 m
l) and potassium carbonate (6.4 g) were added, and the mixture was added at room temperature to 74
Stirred for hours. The reaction mixture was poured into water and the resulting colorless precipitate was collected by filtration and washed with water. Yield 9.5 g ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.26
(3H, t, J = 7Hz), 2.05-2.25 (2
H, m), 2.52 (2H, t, J = 7 Hz), 4.0
7 (2H, t, J = 6.3Hz), 4.15 (2H,
q, J = 7 Hz), 6.94 (2H, d, J = 9.2)
Hz), 7.58 (2H, d, J = 9.2Hz).

4- (4-amidinophenoxy) butane
The compound of acid ethyl ester / hydrogen iodide (3 g) was dissolved in pyridine (75 ml) -triethylamine (15 ml), and hydrogen sulfide gas was passed through for 30 minutes while cooling with ice. After passing hydrogen sulfide gas at room temperature for 3 hours, the mixture was stirred for 20 hours. After nitrogen gas was passed through the reaction mixture to remove hydrogen sulfide, the mixture was concentrated under reduced pressure. The residue was dissolved in acetone (60 ml), methyl iodide (5 ml) was added, and the mixture was stirred at 50 ° C. for 30 min. After allowing to cool, the reaction mixture was concentrated under reduced pressure. The residue was dissolved in methanol (100 ml), ammonium acetate (2.05 g) was added, and the mixture was added to 70
Stirred at 3.5 ° C. for 3.5 hours. After passing through nitrogen gas, ether was added and the resulting precipitate was collected by filtration. Yield 2.74g. Mass spectrum (SIMS): 251 [M + 1] ^{+ 1} H-NMR (CD ₃ OD) δ (ppm): 1.16
(3H, t, J = 7Hz), 1.95-2.1 (2H,
m), 2.45 (2H, t, J = 7.3 Hz), 4.0
-4.15 (4H, m), 7.05 (2H, d, J =
9.2 Hz), 7.71 (2H, d, J = 9.2H)
z).

4- (4-amidinophenoxy) butane
The acid compound (500 mg) was dissolved in 1N hydrochloric acid (5 ml) -acetic acid (5 ml), and the mixture was stirred at 50 ° C for 8 hr. The reaction mixture was concentrated under reduced pressure and the residue was dried to obtain the target product as a brown solid. Yield 388 mg. Mass spectrum (SIMS): 323 [M + 1] ^{+ 1} H-NMR (CD ₃ OD) δ (ppm): 1.95-
2.1 (2H, m), 2.43 (2H, t, J = 7.3
Hz), 4.08 (2H, t, J = 6.3Hz), 7.
06 (2H, d, J = 9 Hz), 7.71 (2H, d,
J = 9 Hz). (2S) -2- (3,4-dimethoxy) benzenesul
Fonylamino-3-t-butoxycarbonylaminopro
Panic acid ethyl ester The compound (350 mg) in Example 1 was used as a raw material, and the compound was synthesized by the method described in Example 1 using (3,4-dimethoxy) benzenesulfonyl chloride. Yield 438 mg. ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.15
(3H, t, J = 7Hz), 1.42 (9H, s),
3.35-3.55 (2H, m), 3.93 (3H,
s), 3.94 (3H, s), 3.95-4.05 (1
H, m), 4.03 (2H, q, J = 7Hz), 4.9
4 (1H, bs), 5.54 (1H, bd, J = 8H
z), 6.92 (1H, d, J = 9 Hz), 7.29
(1H, d, J = 2Hz), 7.46 (1H, dd, J
= 2,9 Hz)

(2S) -3- (4- (4-amidinov
Enoxy) butanoylamino) -2- (3,4-dimeth)
X) Benzenesulfonylaminopropanoic acid TFA salt compound (500 mg) was added to acetonitrile (5 ml)
Dissolve in methanesulfonic acid (557m
A solution of g) in acetonitrile (2.5 ml) was added, and the mixture was stirred at room temperature for 30 minutes. DMF was added to the reaction mixture under ice cooling.
(6.5 ml), triethylamine (0.81 ml),
Compound (359 mg), HOBT.H ₂ O (213
mg) and WSC.HCl (267 mg) were sequentially added, and the mixture was stirred at room temperature for 22 hours. The reaction mixture was concentrated under reduced pressure. The residue was washed with ether, 1N hydrochloric acid (20 ml) -acetic acid (20 ml) was added, and the mixture was stirred at 60 ° C for 16 hr. After the reaction mixture was concentrated under reduced pressure, the residue was purified by HPLC,
The target product was obtained as a colorless powder. Yield 200 mg. HPLC retention time: 21.9 minutes (column: YMC-ODS 4.6 mmΦ × 250 m)
m, detection: UV 220 nm, eluent: solution A 0.1% T
FA / water, B solution 0.1% TFA / acetonitrile, flow rate: 1 ml / min. , Gradient: B solution concentration 10
% To 1% per minute. ) ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 1.8
-1.95 (2H, m), 2.05-2.2 (2H,
m), 3.0-3.15 (1H, m), 3.2-3.4.
(1H, m), 3.75-3.9 (1H, m), 4.0
4 (2H, bt, J = 8Hz), 7.07 (1H, d,
J = 8 Hz), 7.14 (2H, d, J = 9 Hz),
7.27 (1H, d, J = 2Hz), 7.33 (1H,
dd, J = 2,8 Hz), 7.80 (2H, d, J = 8)
Hz), 7.95-8.05 (2H, m), 8.95
(2H, bs), 9.14 (2H, bs)

Example 3 (2S) -3- (4- (4-amidinophenoxy) butane
Noylamino) -2- (4-methoxy) benzenesulfo
Synthesis of NFA amino acid TFA salt (2S) -3-t-butoxycarbonylamino-2-
(4-Methoxy) benzenesulfonylaminopropanoic acid
Ethyl Ester The compound (300 mg) in Example 1 was used as a starting material, and 4-methoxybenzenesulfonyl chloride was used to synthesize the ethyl ester by the method described in Example 1. Yield 500 mg. ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.15
(3H, t), 1.42 (9H, s), 3.46 (2
H, m), 3.86 (3H, s), 3.85-4.10.
(1H, m), 4.02 (2H, q, J = 7Hz),
4.92 (1H, bs), 5.49 (1H, bd, J =
7.6 Hz), 6.96 (2H, d, J = 8.9H)
z), 7.77 (2H, d, J = 8.9Hz).

(2S) -3- (4- (4-amidinov
Enoxy) butanoylamino) -2- (4-methoxy)
Benzenesulfonylaminopropanoic acid TFA salt was synthesized from the compound (133 mg) in Example 2 and the compound (172 mg) in Example 2 by the method described in Example 2. Yield 100 mg. Mass spectrum (SIMS): 479 [M + 1] ⁺ HPLC retention time: 22.3 minutes (column: YMC-ODS 4.6 mmΦ × 250 m)
m, detection: UV 220 nm, eluent: solution A 0.1% T
FA / water, B solution 0.1% TFA / acetonitrile, flow rate: 1 ml / min. , Gradient: B solution concentration 10
% To 1% per minute. ) ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 1.8
0-2.00 (2H, m), 2.10-2.25 (2
H, m), 3.00-3.20 (1H, m), 3.25.
-3.40 (1H, m), 3.80-3.95 (1H,
m), 3.81 (3H, s), 4.05 (2H, t, J
= 6 Hz), 7.06 (2H, d, J = 9 Hz), 7.
14 (2H, d, J = 8.9 Hz), 7.69 (2H,
d, J = 9 Hz), 7.80 (2H, d, J = 8.9H)
z), 7.90-8.10 (2H, m), 8.87 (2
H, bs), 9.13 (2H, bs).

Example 4 (2S) -3- (5- (4-amidinophenoxy) pen
Tanoylamino) -2- (4-methoxy) benzenesul
Synthesis of TFA Salt of Fonylaminopropanoic Acid Ethyl 5- (4-cyanophenoxy) pentanoate
Ter 4-cyanophenol (5.0 g) in DMF (40 m
l) and dissolved in ethyl 5-bromopentanoate (11.4
g) and potassium carbonate (6.38 g) were added, and the mixture was stirred at room temperature for 2 hours.
Stir for 4 hours. The reaction mixture was poured into water and extracted 3 times with ethyl acetate. The organic layer was washed successively with saturated aqueous sodium hydrogen carbonate solution, water and saturated brine, and dried over anhydrous magnesium sulfate. The desiccant was filtered off and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (silica gel 10
0 g, solvent: hexane-ethyl acetate = 5: 1 to 5:
Purified in 2). The fraction containing the target substance was concentrated under reduced pressure to obtain the target substance. Yield 9.86g. ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.26
(3H, t, J = 7Hz), 1.75-1.95 (4
H, m), 2.39 (2H, t, J = 6.9Hz),
4.02 (2H, t, J = 5.9Hz), 4.14 (2
H, q, J = 7 Hz, 6.93 (2H, d, J = 9H)
z), 7.57 (2H, d, J = 9 Hz).

5- (4-amidinophenoxy) penta
Synthesized from the acid compound (1.30 g) by the method described in and in Example 2. Yield 1.66g. ¹ H-NMR (CD ₃ OD) δ (ppm): 1.6-
1.9 (4H, m), 2.31 (2H, t, J = 7H
z), 4.05 (2H, t, J = 5.9Hz), 7.0
6 (2H, d, J = 9Hz), 7.71 (2H, d, J
= 9 Hz). (2S) -3- (5- (4-amidinophenoxy) pe
N-Tanoylamino) -2- (4-methoxy) benzenes
It was synthesized by the method described in Example 2 from the compound (134 mg) of rufonylaminopropanoic acid TFA salt and the compound (154 mg) in Example 3. Yield 91 mg. Mass spectrum (SIMS): 493 [M + 1] ⁺ HPLC retention time: 24.1 minutes (column: YMC-ODS 4.6 mmΦ × 250 m)
m, detection: UV 220 nm, eluent: solution A 0.1% T
FA / water, B solution 0.1% TFA / acetonitrile, flow rate: 1 ml / min. , Gradient: B solution concentration 10
% To 1% per minute. ) ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 1.5
0-1.80 (4H, m), 2.00-2.15 (2
H, m), 3.00-3.20 (1H, m), 3.20
-3.20 (1H, m), 3.75-3.90 (1H,
m), 3.81 (3H, s), 4.06 (2H, bt,
J = 6 Hz), 7.06 (2H, d, J = 8.9H
z), 7.14 (2H, d, J = 8.9 Hz), 7.6
9 (2H, d, J = 8.9Hz), 7.80 (2H,
d, J = 8.9 Hz), 7.85-8.00 (2H,
m), 8.86 (2H, bs), 9.12 (2H, b
s).

Example 5 Platelet Aggregation Inhibitory Action Test Method Blood was collected from the cubital vein of a healthy male (1/10 volume of 3.8% sodium citrate added), and 1000 rpm at room temperature.
The supernatant obtained by centrifugation at (150 g) for 10 minutes was used to obtain platelet rich plasma (PRP). Add 2 μl of the test compound solution to 200 μl of PRP and add 1000 rp at 37 ° C.
After incubating at m for 2 minutes, 22 μl of platelet aggregation agent ADP (adenosine diphosphate, final 3 μg / ml) was added. The platelet aggregating ability was measured by a nephelometry using a hemattracer (Nikko Bioscience). The test result of the platelet aggregation inhibitory activity was expressed as an IC ₅₀ value, that is, a concentration required to suppress the platelet aggregation reaction by 50%. The test results of the test compound are shown in Table 1.

[0043]

[Table 1]

Example 6 Oral absorbability using a bioassay (1) Drug administration and blood collection A guinea pig was fixed under ether anesthesia and a cannula was inserted into the left carotid artery. Each compound was administered orally (5 ml / kg) (the dose was 3 mg / kg in both doses).
And 120 minutes later, 1 ml citric acid-added blood was collected from the cannula. Blood at room temperature 11,000 rpm (6000
The supernatant obtained by centrifugation at g) for 2 minutes was used as plasma for measurement. (2) Quantitative method Citrated blood was collected from guinea pigs donated by platelets (6 animals), and the supernatant obtained by centrifuging at 1000 rpm for 10 minutes at room temperature was subjected to platelet-rich plasma (PRP). The centrifuged supernatant was used as platelet poor plasma (PPP). Furthermore, the volume of the supernatant obtained by centrifuging PRP at 3000 rpm for 5 minutes was halved, and the platelets were resuspended to prepare 2-fold concentrated PRP. PPP
10 prepared at 3.13-100 ng / ml
Mix 0 μl and 100 μl of 2-fold concentrated PRP, mix 10
The platelet aggregation inhibition rate when 22 μl of 0 μg / ml ADP was added was measured. A calibration curve was prepared by plotting the logarithm of the concentration of each compound and the inhibition rate. Plasma of compound-administered animals prepared at each time point (PP if necessary
100 μl (appropriately diluted with P) and 100 μl of PRP were mixed, and 22 μl of 100 μg / ml ADP was added to measure the platelet aggregation inhibition rate. The compound concentration in plasma was calculated from the platelet aggregation inhibition rate at this time. The platelet aggregation ability was measured by a nephelometry using an aggregometer (Nikko Bioscience).

Compound concentration in plasma of each drug (ng / n
Table 1 shows the change over time in 1).

[Table 2]

Reference Example 1 (2S) -3- (5- (4-amidinophenoxy) pen
Tanoylamino) -2- (2,4,6-trimethyl) be
Synthesis of benzenesulfonylaminopropanoic acid TFA salt was synthesized in the same manner as in Example 4. Yield: 191 mg. Mass spectrum (SIMS): 505 [M + 1] ⁺ HPLC retention time: 30.6 minutes (same conditions as in Example 1) ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 1.4
5-1.80 (4H, m), 1.90-2.10 (2
H, m), 2.23 (3H, s), 2.49 (3H,
s), 2.51 (3H, s), 3.00-3.40 (2
H, m), 3.70-3.90 (1H, m), 4.00.
-4.15 (2H, m), 6.98 (2H, s), 7.
14 (2H, d, J = 8.9 Hz), 7.80 (2H,
d, J = 8.9 Hz), 7.85-8.00 (2H,
m), 8.85 (2H, bs), 9.12 (2H, b
s).

Reference Example 2 (2S) -3- (4- (4-amidinophenoxy) butane
Noylamino) -2- (1-naphthalene) sulfonyla
Synthesis of minopropanoic acid TFA salt It was synthesized in the same manner as in Example 2. Yield 67 mg. Mass spectrum (SIMS): 499 [M + 1] ⁺ HPLC retention time: 27.3 minutes (column: YMC-ODS 4.6 mmΦ × 250 m)
m, detection: UV 220 nm, eluent: solution A 0.1% T
FA / water, B solution 0.1% TFA / acetonitrile, flow rate: 1 ml / min. , Gradient: B solution concentration 10
% To 1% per minute. ) ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 1.7
5-2.10 (4H, m), 3.05-3.20 (1
H, m), 3.25-3.40 (1H, m), 3.75.
-3.90 (1H, m), 4.00 (2H, t, J = 6
Hz), 7.13 (2H, d, J = 8.9 Hz), 7.
55-7.85 (4H, m), 7.80 (2H, d, J
= 8.9 Hz), 8.00-8.25 (4H, m),
8.64 (1H, d, J = 8.3Hz), 9.05 (2
H, bs), 9.10 (2H, bs).

Reference Example 3 (2S) -3- (3- (4-amidinobenzoylami)
No) propanoylamino) -2- (1-naphthalene) s
Synthesis of Rufonylaminopropanoic acid TFA salt was synthesized in the same manner as in Example 1. Yield: 36 mg. Mass spectrum (SIMS): 512 [M + 1] ⁺ HPLC retention time: 21.9 minutes (column: YMC-ODS 4.6 mmΦ × 250 m)
m, detection: UV 220 nm, eluent: solution A 0.1% T
FA / water, B solution 0.1% TFA / acetonitrile, flow rate: 1 ml / min. , Gradient: B solution concentration 10
% To 1% per minute. ) ¹ H-NMR (DMSO-d ₆ ) δ (ppm): 2.0
5-2.28 (2H, m), 3.00-3.50 (4
H, m), 3.85-3.95 (1H, m), 7.58.
-7.72 (3H, m), 7.72-8.14 (8H,
m), 8.20, 8.48 (1H, d (J = 7Hz),
d (J = 8 Hz)), 8.61-8.68 (2H,
m), 9.16 (2H, m), 9.39 (2H, s).

The structures of the compounds obtained in Examples 1 to 4 and Reference Examples 1 to 3 are shown below.

Embedded image

[Chemical 19]

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁶ Identification number Internal reference number FI Technical indication location C07C 311/29 7419-4H C07C 311/29 (72) Inventor Kaneko Sosei Kasuga, Kasuga, Konohana-ku, Osaka City 3-1-198 Sumitomo Pharmaceutical Co., Ltd. (72) Inventor Seiya Horizawa 3-1-198 Kasugade, Konohana-ku, Osaka City Sumitomo Pharmaceutical Co., Ltd.

Claims (4)

[Claims] 1. Formula (1): [In the formula, R ¹ represents a hydrogen atom or an ester group, R ² represents a phenyl group substituted with 1 to 3 same or different lower alkoxy groups, a 1-naphthyl group or a 2-naphthyl group, and A is A _{-CONH -, - OCH 2 -,} - OCH
₂ CH ₂ - or -CH ₂ CH ₂ - represents a. ] An oral therapeutic agent containing a compound represented by the following or a pharmaceutically acceptable salt thereof. 2. The oral therapeutic agent according to claim ¹ , wherein in the formula (1), R ¹ is a hydrogen atom. 3. Formula (2): Formula (3), Formula (4) or Formula (5) The oral therapeutic agent according to claim 1, which comprises a compound represented by: or a pharmaceutically acceptable salt thereof. 4. An orally-administered platelet aggregation inhibitor.
4. The therapeutic agent for oral administration according to 3 above.