JPS6360738B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the formula () (In the formula, R is H or a guanidinino group

The present invention relates to a 4'-(β-amidinoethenyl)phenylbenzoate derivative represented by the following formula, a pharmaceutically usable acid addition salt thereof, and a method for producing the same. An object of the present invention is to provide compounds useful as pharmaceuticals. The compound of the present invention has the formula () (In the formula, R is H or a guanidino group

A benzoic acid derivative represented by [formula] or its reactive intermediate and the formula () It can be produced by reacting with 4-(β-amidinoethenyl)phenol shown in the following. The reactive intermediates referred to here are those obtained by the reaction of benzoic acid derivatives with acid halides, acid anhydrides, dicyclohexylcarbodiimide (DCC), diphenylphosphoryl azide (DPPA), etc. used in normal dehydration condensation reactions. The reaction intermediates are shown below. The compounds of the present invention include trypsin, plasmin,
It has strong inhibitory activity against kallikrein and thrombin, and is a useful compound as an antitrypsin agent, an antiplasmin agent, an antikallikrein agent, and an antithrombin agent. Furthermore, the compound of the present invention has strong C1-esterase inhibitory activity and anti-complement action, and is a useful compound as an anti-complement agent. Compounds of the invention, especially 4'-(β-amidinoethenyl)phenyl-4-guanidinobenzoate

[Formula] Methanesulfonate has very high solubility in water and can be used not only as an oral preparation but also as a liquid preparation such as an injection. The method for producing the compound of the present invention will be described in more detail. In other words, when synthesizing an acid halide as a reactive intermediate, the formula ()' (X is halogen) A benzoyl halide represented by 4-(β-amidinoethenyl)phenol (), preferably an acid addition salt thereof, is added to a solution of dimethylformamide, pyridine, dimethyl sulfoxide, etc. in the presence of a dehydrohalogenating agent. Make it react.
As the dehydrohalogenating agent, inorganic bases such as potassium carbonate, sodium carbonate, sodium hydroxide, etc., and organic bases such as triethylamine, pyridine, dimethylaniline, etc. can be used, with pyridine being preferred. The reaction readily proceeds at a temperature of -30°C to +80°C, but in order to avoid the formation of by-products, it is preferable to carry out the reaction initially under ice cooling and then at room temperature. Further, although the reaction is completed in 2 to 5 hours, it may be allowed to react overnight. After the reaction is completed, the reaction mixture is treated by a conventional treatment method. For example, when pyridine is used as a reaction solvent, a solvent such as ethyl ether or ethyl acetate is added to the reaction solution, and the precipitated solid is recrystallized from an appropriate solvent, such as a mixture of methanol and ethyl ether. The compounds of the present invention can be obtained as acid addition salts. Also, after dissolving this in a small amount of methanol, a saturated sodium bicarbonate solution was added to form a crystalline carbonate, which was further suspended in methanol, and p-
Addition of an acid such as toluenesulfonic acid to dissolve the carbonate and addition of diethyl ether to the resulting solution provides the corresponding acid addition salt. Acids that can be used include pharmaceutically usable hydrochloric acid, sulfuric acid,
Inorganic acids such as phosphoric acid, acetic acid, lactic acid, citric acid,
Organic acids such as methanesulfonic acid, succinic acid, fumaric acid, maleic acid, etc. Another manufacturing method is to dissolve or suspend a benzoic acid derivative () in an organic solvent such as dimethylformamide or pyridine, and then use dicyclohexylcarbodiimide (DCC) or diphenylphosphoryl azide (DPPA), which are commonly used as dehydration condensation agents.
The compound () of the present invention can also be obtained by reacting with an ester activator such as the following, and adding 4-(β-amidinoethenyl)phenol (). For example, when DCC is used as a dehydration condensation agent and pyridine is used as a solvent, the reaction easily occurs at room temperature. The reaction is completed in 3 to 5 hours, but it may be allowed to react overnight. After the reaction is complete, remove the target compound and the by-product dicyclohexylurea (DCU) that precipitates in the reaction solution, dissolve it in water to remove the water-insoluble DCU, and add saturated hydrogen carbonate to the filtrate. () can be obtained as carbonate by adding sodium solution. Carbonates can be made into other acid addition salts by the methods previously described. 4-(β-
Amidineethenyl)phenol () is a new substance and is useful as a raw material for the synthesis of the substance of the present invention. The method for synthesizing 4-(β-amidinoethenyl)phenol () used as a raw material is shown below.
This is just an example of a synthesis method, and the method is not restrictive. ()4-(β-amidinoethenyl)phenol Enzyme inhibitory action The compound of the present invention has trypsin, plasmin,
It has strong inhibitory activity against enzymes such as kallikrein and thrombin, and is effective as an antitrypsin agent for the treatment of pancreatitis, an antiplasmin agent and antikallikrein agent effective for the treatment of bleeding disorders, and an anti-kallikrein agent for the treatment of blood clots. It is a useful compound as an effective antithrombin agent. Enzyme inhibitory activity was determined using the method of Muramatsu et al. [M.
Muramatsu, T.onishi, S.Makino, Y.Hayashi
and S. Fujii, J.Biochem., 58 , 214 (1965)] using the methanesulfonate salt of the compound of the present invention. Table 1 shows the results. The data in the table is
The concentration of compound (ID ₅₀ ) that inhibits the ability of each enzyme to hydrolyze TAME (tosyl arginine methyl ester) by 50% is shown.

[Table] Anti-complement effect The compounds of the present invention and their pharmaceutically usable acid addition salts have strong C1 esterase inhibitory activity, complement hemolysis inhibiting activity, and activation of the complement system based on antigen-antibody reactions, which play important roles. It is said to have a therapeutic effect on falsmanshock. This indicates that it is useful as an anti-complement agent effective for allergic diseases such as nephritis involving complement. The results of these tests are shown below. (1) In vitro test (C1 esterase inhibitory activity,
Complement hemolysis inhibition) Okamura et al.'s method using the methanesulfonate of compound () [K.Okamura, M, Muramatsu and
S. Fujii: Biochem. Biophys Acta, 295 , 252−
257 (1973)] and the method of Baker et al. [BRBaker and
EHEricksou J. Med.Chem,. 12 , 408-414
(1969)] Complement hemolysis inhibition rate (%)
are shown in Table 2.

[Table] (2) Pharmacological test (Forsmanschik) This experiment was performed using the method of IGOfferness et al. (Biochem.
Pharmacol. 27 (14) 1873-1878, 1978). Male Hartley guinea pigs weighing approximately 300 g were used. Minimum amount of hemolysin that can cause shock
0.5ml/guinea pig (commercially available hemolysin,
5000U was intravenously injected and the time until death was measured, and this was used as a control group. In the drug treatment group, compound () methanesulfophosphate was administered intravenously 10 minutes before hemolysin administration, and the time to death (seconds) was determined.
was measured.

【table】

Regarding the experimental results using [Formula], it was clearly observed that the time to death was prolonged in the 3 mg/Kg administration group shown in Table 3. Method of Administration The compounds of the present invention are preferably administered orally, but may also be administered by injection. A compound of the invention can be administered as a single therapeutic agent or as a mixture with other therapeutic agents. Although they may be administered alone, they are generally administered in the form of pharmaceutical compositions. Examples of such compositions include tablets, powders, capsules, syrups, and aqueous solutions. Oral compositions may contain conventional additives such as binders, excipients, lubricants, disintegrants, and wetting agents. Oral solutions may be in the form of aqueous or oily suspensions, solutions, emulsions, syrups, elixirs, etc., or may be provided as a dry syrup for reconstitution with water or other suitable solvent before use. may be done. Said solutions may contain customary additives such as suspending agents, flavorants, diluents or emulsifiers. For injection, it can be used as an aqueous solution or an oily suspension. Dose The compound of the present invention can be administered to mammals (including human patients) from 10 to
It can be administered at an oral dosage of 200 mg/day.
In addition, for intravenous administration, it can be administered at a dosage of 1 to 20 mg/day. However, these numbers are just examples, and the most appropriate dose should be administered to the patient depending on the patient's age, weight, and severity of symptoms. Next, examples of formulations of the compounds of the present invention will be given. Formulation example 1 Capsule Compound () methanesulfonate 100.0mg Lactose 59.0mg Crystalline cellulose 33.4mg Calcium carboxymethyl cellulose
3.6mg Magnesium Stearate 4.0mg Total 200.0mg 2 Fine Granules Compound (2) Methanesulfonate 50.0mg Lactose 249.0mg Mannitol 75.0mg Corn Starch 110.0mg Hydroxypropyl Cellulose 16.0mg Total 500.0mg 3 Injection Compound (2) Methanesulfone Acid salt 5.0mg Distilled water for injection 2ml Make an injection using the usual method. Toxicity Table 4 shows the LD ₅₀ of the methanesulfonate salt of the compound of the present invention.

[Table] Example 1 Synthesis of 4-(β-amidinoethenyl)phenylbenzoate (I,R=H) methanesulfonate 4-(β-amidinoethenyl)phenol ()
Dissolve 500 mg of methanesulfonate in 5 ml of pyridine, and add 300 mg of benzoyl chloride under ice cooling. After stirring overnight at room temperature, ether was added, and after stirring well again, the ether layer was removed by decantation. After repeating this operation once more, the remaining solid substance is recrystallized from a methanol-ether mixture. Yield 98 mg (15%). mp.185−193℃. IR (KBr) cm ^-1 ; 3400-3050 (amidino group) 1730
(ester group). Example 2 4'-(amidinoethenyl)phenyl-4-guanidinobenzoate

[Formula] Synthesis of methanesulfonate 4-guanidinobenzoic acid

[Formula] Dissolve 106 g of hydrochloride in 1.3 g of pyridine and stir under ice cooling. DCC122 to this
g and stirred for 30 minutes, then 4-(β-amidinoethenyl)
127 g of phenol methanesulfonate was slowly added, the resulting clear solution was stirred overnight, and the precipitated solid was filtered off. Soak the solids in 2 parts of water,
Stir thoroughly and filter. Add about 2 liters of saturated sodium chloride water to the filtrate.
Add when cold, stir, filter the precipitated crystals, wash with water,
Wash with acetone and briefly air dry the orange-yellow crystals. Dissolve in 200ml of methanol and add about 100g (2eq) of methanesulfonic acid while stirring while cold. Add ether to precipitate crystals and collect by filtration. This is dissolved in methanol to make carbonate again. A white carbonate was obtained. Dissolve this in 300 ml of ethanol at room temperature, add methanesulfonic acid to make a clear solution, and stir for a while to precipitate crystals. Filter when cold and wash thoroughly with cold ethanol. Yield 45g (18%). mp94~97℃. IR (KBr) cm ^-1 : 3400-3050 (amidino group, guanidino group) 1730 (ester group). NMR (DMSO-d ₆ ) ppm: 2.5 (6H, S, 2MSA)
8.3-10.0 (10H, m, aromatic and double bonds) 10.3-10.9 (4H, b, d. amidino group). Examples 3-6, described below, illustrate the preparation of intermediates. Example 3 Synthesis of acetylcoumaric acid 500 g (4.1 mol) of p-hydroxybenzaldehyde and 1.5 kg of acetic anhydride were added to the eggplant-shaped Corbel from 3.
(3 eq.) and 773 g (2.3 eq) of sodium acetate, and after attaching an air cooling tube, heat in an oil bath at 145°C for 20 hours. Stir occasionally during the reaction to pulverize the solids. The reaction solution becomes a red solution. When hot, boiling water 40
Add to and stir. The precipitated ocher crystals are collected by filtration and dried. Stir this in hot methanol from Step 5 and remove the soluble portion. Concentrate methanol, collect the precipitated crystals by filtration, and air dry. Yield 430g (51%). mp.168
−172℃. IR (KBr) cm ^-1 : 3000~2500 (-COOH), 1740
(ester). Example 4 Synthesis of acetylcoumaric acid chloride 180g of acetylcoumaric acid is suspended in 1.5ml of ethyl acetate, and 240g of phosphorus pentachloride is added to this with stirring. As the reaction progresses, it becomes a brown transparent solution.
After standing at room temperature overnight, it was concentrated under reduced pressure, and the precipitated crystals were collected by filtration and washed with hexane to obtain pale yellow crystals. Yield 180g (92%). Example 5 Synthesis of acetyl coumalic acid amide 200 g of acetyl coumalic acid chloride was suspended in 1.5 g of ethyl acetate, stirred at room temperature, and ammonia gas was sufficiently blown into the mixture. The condition changes and the overall whiteness becomes dull. After standing overnight, the crystals are washed thoroughly with ethyl acetate, then with saturated aqueous NaHCO ₃ , water, and air-dried. Yield: 76.9g (44%). mp150−153℃. IR (KBr) cm ^-1 : 3300, 3150 (NH), 1755, (ester), 1660 (amide) Example 6 Synthesis of 4-(β-amidinoethenyl)phenol methanesulfonate 173 g (0.82 mol) of coumaric acid amide Suspend in 1.5 g of anhydrous methylene chloride and stir at room temperature.
1.0 of an anhydrous methylene chloride solution containing 204 g (1.3 eq) of Meyer Wein's reagent dissolved therein was added dropwise. The reaction solution gradually became a transparent solution. After standing overnight, the brown solution was concentrated to about 300 ml, 1.5 ml of anhydrous methanol was added thereto, and ammonia gas was blown in for 3 hours while stirring at room temperature. (exothermic reaction). After standing overnight, the resulting brown-brown solution is filtered to remove insoluble matter. Add water to this and stir. Yellow crystals begin to precipitate. After filtering, wash with acetone and air dry.
Yield 121g (91%)...free base. Suspend the above free base in 150 ml of methanol,
Add 85 g (1.2 eq) of methanesulfonic acid while stirring while cold. Ether is added to the resulting clear solution to precipitate crystals. Filter this out and air dry. yield
127g (67%). mp.147−149℃. IR (BKr) cm ^-1 : 3350, 3100 (NH, OH), 1670
(C=N).

Claims (1)

[Claims] 1 Formula () A 4'-(β-amidinoethenyl)phenylbenzoate derivative represented by the formula (wherein R represents H or a guanidino group) and a pharmaceutically usable acid addition salt thereof. 2 formula () (In the formula, R represents H or a guanidino group [Formula]) A benzoic acid derivative or its reactive intermediate represented by the formula () 4-(β-amidinoethenyl)phenol represented by the formula () (wherein R represents H or a guanidino group [Formula]) A method for producing a 4'-(β-amidinoethenyl)phenylbenzoate derivative and a pharmaceutically usable acid addition salt thereof. 3 Formula () as active ingredient (wherein R represents H or a guanidino group [Formula]) An enzyme activity inhibitor containing a 4'-(β-amidinoethenyl)phenylbenzoate derivative or a pharmaceutically usable acid addition salt thereof.