JPH0848664A - New guanidinobenzoic acid ester derivative - Google Patents

Abstract

PURPOSE:To obtain a new compound, having inhibiting actions on serine proteases and useful for treating and preventing various diseases such as hemorrhagic diseases, thrombus or pancreatitis. CONSTITUTION:This compound is expressed by formula I [A is (CH2)n or styrene; (n) is 0-5; R<1> is H, a halogen or a lower alkyl; R<2> is (CH2)mCOR<3>, etc.; R<3> is 2-thiazolylamino, etc.; (m) is 2 or 3] and its salts. The compound of formula I is obtained by reacting a compound of formula II with a compound of formula III in the presence of a condensing agent (e.g. dicyclohexylcarbodiimide) in a solvent (e.g. pyridine). An inhibitor of serine proteases containing the compound of formula I can orally or parenterally and stably be administered and the daily dose thereof is 10-500mg at a time in the case of oral administration.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel guanidinobenzoic acid ester derivative useful as a drug having serine protease inhibitory activity.

[0002]

BACKGROUND OF THE INVENTION Various serine proteases exist in the body, and it is considered that when these enzymes are abnormally activated by some factors, diseases such as inflammation, pain, allergies, blood abnormalities and tissue destruction are caused. Has been. It is conventionally known that various guanidinobenzoic acid derivatives inhibit the activities of serine proteases trypsin, plasmin, thrombin, elastase and the like and can be used clinically. As a typical example, JP-A-51-13864
No. 2, JP 62-103058 A, JP 6
2-155253, JP-A-63-165357, JP-A-4-46148, and JP-A-4-159.
The compounds described in Japanese Patent No. 261 and the like are known. However, nothing is disclosed regarding the guanidinobenzoic acid ester derivative of the present invention.

[0003]

Therefore, the present invention aims to develop a serine protease inhibitor having a sufficient serine protease inhibitory action and less side effects, and is useful for various diseases mediated by serine protease. Another object of the present invention is to provide a novel guanidinobenzoate derivative and a serine protease inhibitor containing the compound.

[0004]

The guanidinobenzoic acid ester derivative of the present invention has the following general formula (I):

[Chemical 3] [In the formula, A is (CH ₂ ) _n (wherein n is an integer of 0 to 5) or a styrene group, R ¹ is a hydrogen atom, a halogen atom, a lower alkyl group, and R ² is-(CH ₂ ) _m COR
³ (In the formula, m is an integer of 2 or 3, R ³ is 2-thiazolylamino group, (4,5-dimethylthiazol-2-yl) amino group, 3,5-dichloroanilino group, 2-pyridazylamino group , (5-methylisoxazole-3-yl) amino group, piperidino group, 2-methoxy shows a carbonyl anilino _{group), - (CH 2) m} COOR 4 ( wherein, m is as defined above, R ⁴ represents a hydrogen atom, a lower alkyl group, a substituted or unsubstituted benzyl group), —NH
(CH ₂ ) _n COR ³ (In the formula, n is an integer of 1 to 5;
³ has the same meaning as _{above), - NH (CH 2)} n CO
OR ⁴ (in the formula, n and R ⁴ represent the same meaning as described above), −
^{NHCH - (- R 5) -COOR} 4 ( wherein the same meaning as R ⁴ is the, R ⁵ represents a substituted or unsubstituted benzyl group, a methoxycarbonylmethyl _{group), - NH-C 6 H} 4
-(CH ₂ ) _p COOR ⁴ (in the formula, p represents an integer of 0 or 1, and R ⁴ has the same meaning as described above)], and a guanidinobenzoic acid ester derivative and salts thereof. It is about.

The above general formula (I) will be specifically described. The halogen atom is fluorine, chlorine, bromine, iodine, and the lower alkyl group is methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,
The substituent of the substituted benzyl group is an alkyl group having 1 to 6 carbon atoms such as tert-butyl, n-pentyl, n-hexyl and the like, and the substituent of the substituted benzyl group is a halogen atom, a nitro group, a lower alkyl group, a hydroxy group, methoxy, ethoxy, Propoxy, iso-
Propoxy, n-butoxy, iso-butoxy, ter
It means a lower alkoxy group having 2 to 6 carbon atoms such as t-butoxy. In addition, salts thereof include inorganic acid salts such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, and acetic acid, maleic acid, oxalic acid, fumaric acid, succinic acid, malonic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, and the like. Organic acid salts, but are not limited to these.

When the compound represented by the general formula (I) is used as a medicine, as it is or together with a known excipient,
Oral systemically or systemically as appropriate dosage forms such as tablets, capsules, powders, fine granules, suppositories, injections, ointments, gels, creams, reservoir patches, aerosols, etc. Alternatively, it can be stably administered parenterally. The dose is appropriately determined according to the symptoms, age, sex, etc. of the administration subject, but when orally administered to an adult,
Compound (I) or a salt thereof in a single dose of 10 to 500 mg
It is preferable to administer about 1 to several times a day.

Next, a method for producing the compound of the present invention will be described. The compound of the present invention can be obtained in good yield by the method described below, but the scope of the present invention is not limited thereto. Manufacturing method 1

[Chemical 4] (In the formula, A, R ¹ and R ² have the same meanings as described above.) The compound represented by the general formula (II) and the compound represented by the general formula (III) are treated with a condensing agent in a suitable solvent. By reacting in the presence, the compound represented by the general formula (I) can be obtained. Here, the reaction solvent is not particularly limited as long as it is a solvent that does not participate in the reaction, such as pyridine, dimethylformamide, chloroform, dichloromethane, carbon tetrachloride, benzene, toluene, diethyl ether, dioxane, tetrahydrofuran, ethyl acetate, dimethyl sulfoxide, and the like. . As the condensing agent, for example, dicyclohexylcarbodiimide, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, diphenylphosphoryl azide, N, N′-carbodiimidazole and the like can be used, but the condensing agent is not limited thereto. is not.

Manufacturing method 2

Embedded image (In the formula, A, R ¹ and R ² have the same meanings as described above, and H
al means a halogen atom. ) A compound represented by the general formula (I) is obtained by reacting a compound represented by the general formula (IV) with a compound represented by the general formula (III) in the absence of a solvent or a suitable solvent in the presence of a base catalyst. Can be obtained. Here, the reaction solvent is not particularly limited as long as it is a solvent that does not participate in the reaction, such as chloroform, dichloromethane, carbon tetrachloride, tetrahydrofuran, dioxane, dimethylformamide, dimethylsulfoxide. As the base catalyst, pyridine, triethylamine, tri-n-propylamine, N-methylmorpholine and the like can be used,
It is not limited to these. The compounds represented by the general formulas (II), (III) and (IV) used in the above production method can be produced by a known combination of reactions.

[0009]

The present invention will be described in more detail with reference to the following examples. However, the present invention is not limited to these examples.

Example 1 Pyridyl 4-guanidinobenzoic acid hydrochloride (1.08 g)
Solution (20 ml) in dicyclohexylcarbodiimide
(Hereinafter DCC, abbreviated as 1.03 g) is added at -15 ° C, and 2
After stirring for 0 minutes, N- (2-thiazolyl) -4-hydr
Add Roxybenzamide (1.10g) and stir at 0 ° C overnight.
Allowed to stir. After the reaction is complete, the insoluble matter is filtered off and the filtrate is saturated.
Aqueous sodium hydrogen carbonate solution was added, and the formed crystals were collected by filtration and washed with water.
And sequentially washed with acetone and dried. Methano crystals
Suspended in water, added methanesulfonic acid (0.5 g),
Insoluble matter was filtered off, acetone was added to the filtrate, and the precipitated crystals were collected.
It was dried to obtain a compound having the following structure (1.34 g). Melting point
Was 143-148 ° C. IR: 3150, 1740, 1684, 1574, 12
09,1060 cm^-1 Elemental analysis value (C₁₈H_FifteenN 5 O 3S / 2CH 3SO 3H / 2H Calculated value C: 39.40 H: 4.46 N: 11.49 Actual value C: 39.48 H: 4.27 N: 11.42

[Chemical 6]

Example 2 DCC (0.99 g) was added to a solution of 4-guanidinobenzoic acid hydrochloride (1.04 g) in pyridine (15 ml) -dimethylformamide (5 ml) at -15 ° C, and the mixture was stirred for 20 minutes. N- (2-thiazolyl) -3- (4-hydroxybenzoyl) propanamide (1.33 g) was added,
Let stir at 0 ° C. overnight. After the reaction is completed, the insoluble matter is filtered off,
Saturated aqueous sodium hydrogencarbonate was added to the filtrate, and the resulting crystals were collected by filtration, washed successively with water and acetone and dried. The crystals were suspended in ethanol, and methanesulfonic acid (0.5
g) was added, the insoluble matter was filtered off, the filtrate was left overnight at 4 ° C., the precipitated crystals were dried, and the compound of the following structure (0.3
5 g) was obtained. The melting point was 182-186 ° C. IR: 3382, 1723, 1686, 1576, 11
93,1044 cm ^-1 Elemental analysis value (as C ₂₁ H ₁₉ N ₅ O ₄ S.2CH ₃ SO ₃ H.1 / 2H ₂ O) Calculated value C: 43.25 H: 4.42 N: 10.97 Actual measurement Value C: 43.19 H: 4.40 N: 10.86

[Chemical 7]

Example 3 4-guanidinobenzoic acid methanesulfonate (2.75)
g) in pyridine (50 ml) in DCC (2.06
g) was added at -15 ° C and stirred for 20 minutes, and then 3- (4
4-Nitrobenzyl (hydroxybenzoyl) propionate (3.29 g) was added, and the mixture was stirred at 0 ° C. overnight. After completion of the reaction, the insoluble matter was filtered off, saturated aqueous sodium hydrogencarbonate was added to the filtrate, and the resulting crystals were collected by filtration, washed successively with water and acetone, and dried. Suspend the crystals in methanol,
Methanesulfonic acid (1.0 g) was added, the insoluble material was filtered off, and the filtrate was concentrated under reduced pressure. The concentrate was purified by silica gel chromatography (chloroform: methanol = 5: 1) to obtain a compound (1.6 g) having the following structure. The melting point was 78-82 ° C. IR: 3370, 1740, 1686, 1603, 12
09,1166 cm ^-1 Elemental analysis value (as C ₂₅ H ₂₂ N ₄ O ₇ .CH ₃ SO ₃ H.1 / 2H ₂ O) Calculated value C: 52.43 H: 4.57 N: 9.41 Measured value C: 52.30 H: 4.68 N: 9.28

Embedded image

Example 4 4- [3- (4-nitrobenzyloxycarbonyl) propanoyl] phenyl-4-guanidinobenzoate carbonate (3.9 g) synthesized in Example 3 was mixed with acetic acid (70 m).
5% -Pd / C (0.4 g) was added to a mixed solution of 1), methanol (50 ml) and water (30 ml), and hydrogenation was carried out under normal pressure for 3 hours. After completion of the reaction, the insoluble matter was filtered off, acetone was added to the filtrate, and the generated crystals were collected by filtration and dried. The crystals were suspended in a dimethylformamide-acetone solution, methanesulfonic acid (1.0 g) was added, the insoluble matter was filtered off, ether was added to the filtrate, and the precipitated crystals were dried to give a compound of the following structure (0. 45 g) was obtained. Melting point is 2
It was 12-214 degreeC. IR: 3168, 1738, 1709, 1673, 15
76,1199 cm ⁻¹ Elemental analysis value (as C ₁₈ H ₁₇ N ₃ O ₅ .CH ₃ SO ₃ H) Calculated value C: 50.55 H: 4.69 N: 9.31 Measured value C: 50.41 H : 4.64 N: 9.36

[Chemical 9]

Examples 5 to 30 The compounds of the present invention shown in Tables 1 to 3 were synthesized according to the methods of Examples 1 to 4. Melting points indicate results with methanesulfonate.

[0015]

[Table 1]

[0016]

[Table 2]

[0017]

[Table 3]

In order to prove that the compound of the present invention exhibits serine protease inhibitory activity, the experimental method and the result of kallikrein and plasmin inhibitory activity are shown below.

Experiment on Kallikrein Inhibitory Activity Human plasma kallikrein and HD-Pro-Phe
-Arg-p-nitroanilide. That is, various specimens and enzyme solutions were prepared with 0.05 M Tris-hydrochloric acid buffer solution (pH 7.8), and the reaction mixture solution was mixed with 3
Incubated at 7 ° C for 4 minutes. The reaction was stopped by adding 15% acetic acid, and the absorbance was measured at 405 nm. The test results were expressed as 50% inhibitory concentration (IC ₅₀ ).

Experiment on plasmin inhibitory activity Human plasmin and HD-Val-Leu-Lys-
Performed with p-nitroanilide. That is, 0.0
Various samples and enzyme solutions were prepared with 5 M Tris-hydrochloric acid buffer solution (containing 110 mM NaCl, pH 7.4),
These reaction mixtures were incubated at 37 ° C for 4 minutes. The reaction was stopped by adding 15% acetic acid, and the absorbance was measured at 405 nm. The test results are expressed as IC ₅₀ .

Results The results of the inhibitory activity (IC ₅₀ ) are shown in Table 4 below.

[Table 4]

[0022]

The compound [1] of the present invention has an excellent serine protease inhibitory activity. Therefore, it can be expected to be used as a therapeutic agent or a preventive agent for various diseases such as hemorrhagic diseases mediated by serine protease, thrombosis, pancreatitis, etc., and a useful medicine can be provided.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A61K 31/50 C07C 309/04 7419-4H C07D 237/08 261/14 277/44 295/18 Z (72) Inventor Noriyuki Fujimoto 408 Tashiro Daikancho, Tosu City, Saga Prefecture, Hisamitsu Pharmaceutical Co., Ltd. (72) Inventor, Kazuhisa Takeda 408, Tashiro Daikanmachi, Tosu City, Saga Prefecture

Claims (2)

[Claims] 1. A compound of the general formula (I) [In the formula, A is (CH ₂ ) _n (wherein n is an integer of 0 to 5) or a styrene group, R ¹ is a hydrogen atom, a halogen atom, a lower alkyl group, and R ² is-(CH ₂ ) _m COR
³ (In the formula, m is an integer of 2 or 3, R ³ is 2-thiazolylamino group, (4,5-dimethylthiazol-2-yl) amino group, 3,5-dichloroanilino group, 2-pyridazylamino group , (5-methylisoxazole-3-yl) amino group, piperidino group, 2-methoxy shows a carbonyl anilino _{group), - (CH 2) m} COOR 4 ( wherein, m is as defined above, R ⁴ represents a hydrogen atom, a lower alkyl group, a substituted or unsubstituted benzyl group), —NH
(CH ₂ ) _n COR ³ (In the formula, n is an integer of 1 to 5;
³ has the same meaning as _{above), - NH (CH 2)} n CO
OR ⁴ (in the formula, n and R ⁴ represent the same meaning as described above), −
^{NHCH - (- R 5) -COOR} 4 ( wherein the same meaning as R ⁴ is the, R ⁵ represents a substituted or unsubstituted benzyl group, a methoxycarbonylmethyl _{group), - NH-C 6 H} 4
-(CH ₂ ) _p COOR ⁴ (in the formula, p represents an integer of 0 or 1, and R ⁴ has the same meaning as described above)], and a guanidinobenzoic acid ester derivative and salts thereof. 2. A compound represented by the general formula (I): [In the formula, A is (CH ₂ ) _n (wherein n is an integer of 0 to 5) or a styrene group, R ¹ is a hydrogen atom, a halogen atom, a lower alkyl group, and R ² is-(CH ₂ ) _m COR
³ (In the formula, m is an integer of 2 or 3, R ³ is 2-thiazolylamino group, (4,5-dimethylthiazol-2-yl) amino group, 3,5-dichloroanilino group, 2-pyridazylamino group , (5-methylisoxazole-3-yl) amino group, piperidino group, 2-methoxy shows a carbonyl anilino _{group), - (CH 2) m} COOR 4 ( wherein, m is as defined above, R ⁴ represents a hydrogen atom, a lower alkyl group, a substituted or unsubstituted benzyl group), —NH
(CH ₂ ) _n COR ³ (In the formula, n is an integer of 1 to 5;
³ has the same meaning as _{above), - NH (CH 2)} n CO
OR ⁴ (in the formula, n and R ⁴ represent the same meaning as described above), −
^{NHCH - (- R 5) -COOR} 4 ( wherein the same meaning as R ⁴ is the, R ⁵ represents a substituted or unsubstituted benzyl group, a methoxycarbonylmethyl _{group), - NH-C 6 H} 4
-(CH ₂ ) _p COOR ⁴ (in the formula, p represents an integer of 0 or 1 and R ⁴ has the same meaning as described above)] and contains a guanidinobenzoate derivative and salts thereof as an active ingredient. Serine protease inhibitor.