JPH0669379B2 - Process for producing cyclohexanecarboxylic acid derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION <Field of Industrial Application> The present invention relates to trans-4-aminomethylcyclohexanecarboxylic acid-4 ′-(2 ″ -carboxy), which has excellent effects as a therapeutic agent for gastritis / gastric ulcer and an anti-brasmin agent. Ethyl) phenyl ester (hereinafter referred to as CEP ester)
The present invention relates to a novel and industrially advantageous production method.

<Prior Art> As a method for producing a CEP ester, a method is known in which a protecting group is eliminated from a raw material compound in which a terminal carboxyl group is protected. The method includes (1) a method by catalytic reduction (Japanese Patent Publication No. 46-19950 and Japanese Patent Publication No. 52-48978),
(2) Method by acid-alkali hydrolysis (JP-A-52-17)
447 and JP-A-53-56642) and (3) Lewis acid-
A method using a nucleophilic reagent (JP-A-56-167648) can be mentioned.

However, these methods have the following drawbacks.

That is, in the method (1), an expensive catalyst such as palladium is required, and since hydrogen gas must be reacted in a pressurized state, there is a safety problem, and a special device is required. . In the method (2), the ester at the center of the molecule of the CEP ester is easily hydrolyzed and the number of raw material compounds is complicated. In the method (3), the post-treatment of the metals used is complicated.

<Problems to be Solved by the Invention> The present inventors have completed the present invention as a result of intensive studies to solve the above-mentioned drawbacks.

<Structure of Invention> The present invention has the general formula (I) (In the formula, R represents a benzyl group which may have a substituent, a phenyl group or a lower alkyl group.) A compound having the ability to convert into a CEP ester and belonging to the genus Aspergillus or Mucor. The present invention relates to a method for producing a CEP ester or a salt thereof, which comprises treating the compound of the above formula (I) or a salt thereof in the presence of cells, a culture, or a fraction thereof.

Examples of the substituent of the benzyl group in the formula (I) include a halogen atom, a lower alkyl group and a nitro group.

Further, as the substituent R in the formula (I), a benzyl group is preferable.

Examples of the salt of the compound of formula (I) include acid addition salts with inorganic acids such as hydrochloric acid and sulfuric acid. As the salt of CEP ester, an acid addition salt with an inorganic acid such as hydrochloric acid or sulfuric acid may be mentioned.

Preferred species of Aspergillus microorganisms include Aspergillus nigar and Aspergillus oryzae,
Examples of preferable microorganisms of the genus Mucor include Mucor japonicus.

Examples of the culture include those obtained by culturing the above microorganism in an appropriate medium.

Examples of the fraction of the microorganism include cells or cultures extracted with water or an appropriate buffer, a culture filtrate when cultured in a fluid medium, ammonium sulfate or alcohol in the extract or filtrate. The precipitate obtained by the addition of the precipitate and the extract or filtrate obtained by fractionation using gel filtration, ultrafiltration, ion exchange chromatography, etc., and crushed bacterial cells and the crushed product described above. What was fractionated using can be mentioned. Some of these fractions are commercially available as enzyme preparations. Examples of the enzyme preparation include lipase M-AP, pectinase PL, pectinase G, cellulase (I), cellulase (II), cellulase, sumizyme AC and the like, preferably sumezyme AC and ribase M-AP. In general, it is preferable to carry out the reaction in the presence of a commercially available enzyme preparation for the sake of simplicity.

The reaction of the present invention is usually carried out with water or a suitable buffer pH of about 4-5.
5 to 20 to 40 ° C, preferably 25 to 30 ° C.
It is carried out for up to 20 hours, preferably 10 to 15 hours. Examples of the buffer solution include phosphoric acid and citrate buffer solutions. A suitable concentration of the compound of formula (I) as a raw material in the reaction solution is usually 8% (w / v) or less. When the reaction is carried out in the presence of a commercially available enzyme preparation, the enzyme preparation is usually used in an amount of about 1/50 parts by weight per 1 part by weight of the compound of the formula (I) as a raw material.

After completion of the reaction, if the pH of the reaction solution is adjusted to around 7, the CEP ester will precipitate, and this is collected by filtration. Then, the obtained crystals are washed with water or dissolved in a mixed solution of water and isopropyl alcohol or an aqueous solution having a pH of about 2.5, treated with activated carbon or a nonionic porous polymer resin, and then crystallized with water or the like. High-purity CEP ester can be obtained by precipitation.

<Effects of the Invention> The production method of the present invention, under mild conditions, does not involve decomposition of the central phenyl ester of the CEP ester, and the target CE
The P ester can be obtained with high purity and high yield.
Further, the production method of the present invention does not require a special device in operation, is excellent in safety, and is advantageous in terms of post-treatment.
Therefore, the manufacturing method of the present invention is excellent as an industrial manufacturing method.

Next, the present invention will be described with reference to Reference Examples and Examples, but the present invention is not limited thereto.

In the examples, trans-4-aminomethylcyclohexanecarboxylic acid-4 '-(2 "-carboxyethyl) phenyl ester is CEP ester, and its hydrochloride is C
It is referred to as EP ester hydrochloride.

Reference Example 1 250 g of wheat flour was suspended in 250 ml of water. After sterilizing this suspension, inoculate the cells of Aspergillus niger,
It was cultured at 30 ° C for 4 days. After culturing, add distilled water 2 to 30 ℃
After stirring for 3 hours, the mixture was filtered. The filtrate was centrifuged and the supernatant was concentrated by ultrafiltration to obtain 200 ml of concentrated liquid.

Reference Example 2 Using Aspergillus oryzae, the cells were cultured and treated in the same manner as in Reference Example 1 to obtain 200 ml of a concentrated solution.

Reference Example 3 Corn steep liquor 15 g, soluble starch 5 g, soybean flour 5
After suspending g, potassium dihydrogen phosphate 1 g, and potassium chloride 0.25 g in water, the total amount was adjusted to 500 ml while adjusting the pH to 5. After this suspension was sterilized, Mucor Yavonix cells were inoculated and cultured at 26 ° C. for 46 hours with shaking. After filtering the culture broth, ammonium sulfate was added to 500 ml of the filtrate to 70% saturation, and the mixture was centrifuged. Distilled water (50 ml) was added to the obtained precipitate, and dialyzed according to a conventional method to obtain 100 ml of an aqueous solution.

Example 1 48 g of trans-4-aminomethylcyclohexanecarboxylic acid-4 '-(2 "-benzyloxycarbonylethyl) phenyl ester hydrochloride (hereinafter abbreviated as compound A) was suspended in 1200 ml of water, and Sumizyme AC (new 0.96 g (manufactured by Nippon Kagaku Kogyo Co., Ltd.) was added, and the mixture was stirred at 30 ° C. for 15 hours.
The pH was kept at 4-4.5 with N sodium hydroxide.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 99.5%.

The reaction solution was adjusted to pH 7 and the precipitation result was collected by filtration. After washing and drying, 33.0 g of CEP ester (yield 97.4%) was obtained.

The obtained CEP ester was suspended in an isopropyl alcohol-water mixed solution, adjusted to pH 2.5 with hydrochloric acid, and treated with activated carbon. The clear filtrate was concentrated under reduced pressure, and the precipitated crystals were collected by filtration and dried to obtain CEP ester hydrochloride (35.7 g, yield 94.1%).

Thin-layer chromatography, liquid chromatography, IR, and NMR of this product were completely consistent with those of the standard product. The purity of this product was 99.9% when examined by liquid chromatography.

Example 2 The same reaction as in Example 1 was carried out except that 4.8 g of Pectinam PL was used instead of 0.96 g of Sumizyme AC.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 98.7%.

The reaction mixture was treated as in Example 1 to give CEP ester hydrochloride 33.
I got 7g.

Example 3 To 24 g of Compound A, 600 ml of water and 80 ml of the concentrate obtained in Reference Example 1 were added, and the reaction system was maintained at pH 4 to 4.5 at 30 ° C.
Stir for 15 hours. When the reaction solution was analyzed by high performance liquid chromatography, the production rate of CEP ester was 99.0%.

The obtained reaction solution was treated in the same manner as in Example 1 to obtain 17.1 g of CEP ester hydrochloride.

Example 4 The same reaction as in Example 3 was carried out using the concentrated liquid obtained in Reference Example 2.

When the reaction solution was analyzed by high performance liquid chromatography
The production rate of CEP ester was 98.6%.

The obtained reaction solution was treated in the same manner as in Example 1 to obtain CEP ester hydrochloride (16.4%).

Example 5 To 24 g of Compound A, 600 ml of 0.2 M citrate buffer (pH 5) and 6 g of Rebase M-AP-10 (manufactured by Amano Pharmaceutical Co., Ltd.) were added, and the mixture was added at 30 ° C.
The mixture was stirred for 15 hours.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 99.7%.

Next, the reaction solution was adjusted to pH 7, and the precipitated crystals were collected by filtration. The obtained crystals were suspended in 1,900 ml of water, adjusted to pH 2.8 with hydrochloric acid, and treated with activated carbon. 35 ml of a nonionic porous polymer resin (Diaion HP-10 (manufactured by Mitsubishi Kasei)) was added to the filtrate, and the mixture was stirred and filtered. The filtrate was concentrated under reduced pressure, and the precipitated crystals were collected by filtration and dried to obtain 16.9 g of CEP ester hydrochloride.

Example 6 To 24 g of Compound A, 600 ml of 0.2 M citrate buffer (pH 5) and 25 ml of the aqueous solution obtained in Reference Example 3 were added, and the mixture was stirred at 30 ° C. for 15 hours.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 98.5%.

The resulting reaction solution was post-treated in the same manner as in Example 5 to obtain 16.1 g of CEP ester hydrochloride.

Example 7 Trans-4-Aminomethylcyclohexanecarboxylic acid-4 '-(2 "-ethoxycarbonylethyl) phenyl ester hydrochloride 18.5 g 0.05 M citrate buffer (pH 4.5) 4
60 ml and Sumizyme AC 18.5 g were added, and the mixture was stirred at 30 ° C. for 15 hours.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 96.5%.

The obtained reaction solution was post-treated in the same manner as in Example 1 to obtain 14.7 g of CEP ester hydrochloride.

Example 8 trans-4-Aminomethylcyclohexanecarboxylic acid-4 '-(2 "-methoxycarbonylethyl) phenyl ester hydrochloride 17.8 g 0.05% citrate buffer (pH 4.5)
1.780 ml and Sumiteam AC 17.8 g were added, and the mixture was stirred at 30 ° C. for 15 hours.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 95.2%.

The obtained reaction solution was post-treated in the same manner as in Example 1 to obtain CEP ester hydrochloride (14.3 g).

Example 9 trans-4-Aminomethylcyclohexanecarboxylic acid-4 '-(2 "-phenoxycarbonylethyl) phenyl ester hydrochloride 20.9 g in 0.05M citrate buffer (pH 4.
5) 2,000 ml and 6 g of Sumizyme AC were added, and the mixture was stirred at 30 ° C for 15 hours.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 98.6%.

The obtained reaction solution was post-treated in the same manner as in Example 1 to obtain 14.0 g of CEP ester hydrochloride.

Example 10 trans-4-Aminomethylcyclohexanecarboxylic acid-4 '-(2 "-p-nitrobenzyloxycarbonylethyl) phenyl ester hydrochloride 23.2 g, 0.05M citrate buffer (pH 4.5) 2,320 ml and Sumizyme AC23.2g was added and it stirred at 30 degreeC for 15 hours.

When the reaction completed liquid was analyzed by high performance liquid chromatography, the production rate of CEP ester was 95.1%.

The obtained reaction solution was post-treated in the same manner as in Example 1 to obtain 13.9 g of CEP ester hydrochloride.

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Claims (1)

[Claims] 1. A formula (I) (In the formula, R represents a benzyl group which may have a substituent, a phenyl group or a lower alkyl group.) A compound represented by trans-4-aminomethylcyclohexanecarboxylic acid-4 '-(2 "-carboxyethyl) ) Treating the compound represented by formula (I) or a salt thereof in the presence of a microorganism, a culture, or a fraction thereof of a microorganism belonging to the genus Aspergillus or the genus Mucor having the ability to convert to a phenyl ester. For producing trans-4-aminomethylcyclohexanecarboxylic acid-4 '-(2 "-carboxyethyl) phenyl ester and salts thereof