JPH0574349B2 - - Google Patents

Description

[Detailed description of the invention]

<Industrial application field> The present invention is based on the formula

The present invention relates to a method for producing optically active 4-hydroxy-2-cyclopentenone represented by the following formula: <Prior Art> Optically active 4-hydroxy 2-cyclopentenone is useful as an intermediate for pharmaceuticals, agricultural chemicals, etc., and especially as a raw material for prostaglandin, which has various pharmacological actions such as anti-ulcer action and thrombolytic action. For this purpose, optically active 4-hydroxy-2-cyclopentenone having R coordination is particularly useful, but recently S
Coordination compounds are also known to be useful, and when used for such pharmaceutical purposes, particularly high optical purity is required. Conventionally, such optically active 4-hydroxy-
As a method for obtaining 2-cyclopentenone, a method of hydrolyzing acetate ester of optically active 4-hydroxy-2-cyclopentenone ester having R coordination using wheat germ lipase enzyme (Tokuko Sho)
No. 55-34677) is known, but there is no known method using other esters as raw materials. By the way, this method, which uses wheat germ lipase, requires a long hydrolysis time of about 48 hours, and due to the long hydrolysis time, racemization simultaneously proceeds during the reaction, resulting in There was a problem in that the optical purity of the optically active 4-hydroxy-2-cyclopentenone obtained was reduced. <Problems to be Solved by the Invention> Based on the above, the present inventors have developed a method for efficiently hydrolyzing optically active cyclopentenone esters in a short period of time without producing a racemate, with a high yield. As a result of studies to produce 4-hydroxy-2-cyclopentenone with high optical purity, it was discovered that the above object could be achieved only when hydrolysis was carried out using a very specific enzyme, and the present invention It came to this. <Means for solving the problems> That is, the present invention solves the general formula

[Chemical formula] (In the formula, R represents a saturated or unsaturated aliphatic hydrocarbon group having 2 to 12 carbon atoms that may be substituted with halogen. * represents an asymmetric carbon atom) An optically active 4-hydroxy-2-cyclopene characterized by hydrolyzing active cyclopentenone esters using an enzyme having esterase activity derived from a microorganism belonging to the genus Arthrobacter, Candeida, or Pseudomonas. This provides a method for manufacturing Tenon. In the present invention, optically active cyclopentenone esters as raw materials are optically active 4-hydroxy-2-
After converting cyclopentenone into a sulfonic acid ester, it is treated with a saturated or unsaturated carboxylic acid salt that may be substituted with a halogen, resulting in an inversion, that is, a configuration opposite to the original configuration. It can be produced as an optically active cyclopentenone ester.

[Chemical formula] Such optically active cyclopentenone esters include, for example, propionic acid ester, n-
Butyric acid ester, isobutyric acid ester, n-valeric acid ester, isovaleric acid ester, pivalic acid ester, methyl ethyl acetate, n-caproic acid ester, isocaproic acid ester, β-
Examples include methylvaleric acid ester, t-butyl acetate, diethyl acetate, methyl-n-propyl acetate, methylisopropyl acetate, 2-methylbutane-2-carboxylic acid ester, heptanoic acid ester, caprylic acid ester, etc. . The hydrolysis reaction in the present invention is carried out using an enzyme having esterase activity derived from a microorganism belonging to the genus Arthrobacter, Pseudomonas, or Candeida. Here, the above-mentioned esterase means esterase in a broad sense including lipase. The above-mentioned microorganisms are usually cultured by liquid culture according to a conventional method, thereby obtaining a culture solution. Furthermore, some of these microbial-derived esterases are commercially available and can be easily obtained. Specific examples of such commercially available esterases include lipase from the genus Pseudomonas [Amano P (manufactured by Amano Pharmaceutical Co., Ltd.]), lipase from Candida cylindratsuhi [Lipase MY
(manufactured by Meito Sangyo)] and Arthrobacter lipase (manufactured by Shin Nihon Kagaku). The esterase used in this reaction is an enzyme obtained from microorganisms, and its usage forms include purified enzymes, crude enzymes, enzyme-containing substances, microbial culture fluids, cultures, bacterial cells, culture oral fluids, and their use. It can be used as needed in various forms such as treated products, and enzymes and microorganisms can be used in combination. Alternatively, it can also be used as an immobilized enzyme or immobilized bacterial cells immobilized on a resin or the like. In the hydrolysis reaction of optically active cyclopentenone esters, a mixture of the raw material optically active cyclopentenone esters and the above enzyme or microorganism,
This is usually done in a buffer solution by vigorous stirring. As the buffer, commonly used buffers with inorganic acid groups such as sodium phosphate and potassium phosphate, buffers with organic acid salts such as sodium acetate and sodium citrate, etc. are used, and the pH thereof is usually PH= 5 to 8 are preferred. Concentration is usually 0.05~
2M, preferably in the range of 0.05-0.5M. The reaction temperature is usually 10-60°C. The reaction time is preferably within 10 hours, particularly within 6 hours, and can be adjusted by adjusting the amount of enzyme used. After the hydrolysis reaction is completed, in order to separate the hydrolysis product from the reaction solution, the hydrolysis reaction solution is extracted with a solvent such as methyl isobutyl ketone, ethyl acetate, or ethyl ether, and the solvent is distilled off from the organic grass. After that, the concentrated residue can be further distilled, or
This is carried out by a method such as column chromatography, whereby optically active 4-hydroxy-2-cyclopentenone is obtained. <Effects of the Invention> According to the method of the present invention, hydrolysis can be carried out in a short time,
Optically active 4-hydroxy-2-cyclopentenone having high optical purity can be obtained without reducing optical purity. The optically active 4-hydroxy-2-cyclopentenone produced in this manner has extremely little racemization and is extremely advantageous when leading to prostaglandin. Furthermore, by combining the method of the present invention with the above-described method for producing optically active cyclopentenone esters, stereoinversion of optically active 4-hydroxy-2-cyclopentenone becomes easily possible.
The industrial use value will be higher. <Examples> The present invention will be explained below with reference to Examples. Reference Example 1 10 g of 4(S)-hydroxy-2-cyclopentenone (optical purity 97%) and 50 g of dichloromethane were placed in a 4-necked flask equipped with a stirrer, a thermometer, and a dropping funnel.
ml and 12.2 g of pyridine, and 12.9 g of methanesulfonyl chloride was added over 2 hours at -10°C. After incubating at the same temperature for 1 hour, the reaction solution was mixed with water and 2
Wash sequentially with % heavy sodium chloride water and water. The organic layer is dried over magnesium sulfate and then concentrated. The concentrated residue is subjected to silica gel column chromatography using a mixture of toluene and ethyl acetate = 5:3. 16.8 g of methanesulfonic acid ester of 4-(S)-hydroxy-2-cyclopentenone are obtained. α] ²⁵ _D −95.1° (C=1, CHCl ₃ ) n ²⁵ _D 1.4855 (crystallizes if left standing) To 10 g of the obtained methanesulfonic acid ester of 4(S)-hydroxy-2-cyclopentynon, add n - Add 18.73 g of sodium butyrate and 60 ml of hexamethylphosphorotriamide and react at 40-60°C for 6 hours. After the reaction is complete, add the reaction solution to ice and add toluene.
Extract with 150ml. The organic layer is washed successively with 2% aqueous sodium chloride solution, water, 2% aqueous hydrochloric acid, and water, dried over magnesium sulfate, and concentrated under reduced pressure. The concentrated residue was purified by chromatography using a mixture of ethyl acetate and toluene = 1:5 to give 4(R)
8.94 g of n-butyric acid ester of -hydroxy-2-cyclopentenone is obtained. α〕 ²⁵ _D +107゜(C=1, methanol) bp60~60℃/0.3mmHg Reference Example 2 4(S)-Hydroxy- obtained in the same manner as Reference Example 1
Add 16.35 g of sodium propionate and 40 ml of hexamethylphosphorotriamide to 10 g of methanesulfonic acid ester of 2-cyclopentenone,
React for 6 hours at ~60°C. After the reaction was completed, post-treatment was carried out in the same manner as in Reference Example 1 to obtain 4(R).
8.1 g of propionic acid ester of -hydroxy-2-cyclopentenone was obtained. α〕 ²⁵ _D +94.6゜(C=1, methanol) b.p68℃/1mmHg Reference Example 3 4(S)-Hydroxy- obtained in the same manner as Reference Example 1
Add 23.44 g of caproic acid sodium salt and 40 ml of hexamethylphosphorotriamide to 10 g of methanesulfonic acid ester of 2-cyclopentenone.
React for 6 hours at ~60°C. After the reaction was completed, post-treatment was carried out in the same manner as in Reference Example 1 to obtain 4(R).
10.32 g of caproic acid ester of -hydroxy-2-cyclopentenone was obtained. α] ²⁰ _D +110.4 (C=1, methanol) n ²⁰ _D 1.4708 Reference Example 4 4(S)-Hydroxy- obtained in the same manner as Reference Example 1
β-chloropropionate sodium salt to 10 g of menanesulfonic acid ester of 2-cyclopentenone
22.31g and hexamethylphosphorotriamide
Add 40 ml and react at 40-60°C for 6 hours. After the reaction was completed, post-treatment was carried out in the same manner as in Reference Example 1 to obtain 4(R).
10.01 g of β-chloropropionic acid ester of -hydroxy-2-cyclopentenone was obtained. [α] ²⁰ _D +73.2°C (C=1, methanol) n ²⁰ _D 1.4898 Example 1 2 g of n-butyric acid ester of 4(R)-hydroxy-2-cyclopentenone obtained in Reference Example 1, Arthro Bacterium lipase (manufactured by Shin Nippon Chemical) 200mg and
Mix 10 ml of 0.5M phosphate buffer aqueous solution (PH=7) and stir vigorously at 25-30°C for 2.5 hours. After the reaction is completed, Glauber's salt is added to the reaction solution, extracted with methyl isobutyl ketone, and the extract is concentrated. Furthermore, the concentrated residue is further distilled using a micro vacuum distiller to obtain -4
1.11 g of (R)-hydroxy-2-cyclopentenone was obtained. α] ²⁰ _D +93.5°C (C=1, methanol) Optical purity 96.5% Example 2 In place of the Arthrobacter lipase in Example 1, Pseudomonas lipase “Amano P” (manufactured by Amano Pharmaceutical Co., Ltd.) 200 mg 1.03 g of 4(R)-hydroxy-2-cyclopentenone was obtained by carrying out the reaction and post-treatment in the same manner as in Example 1, except that the mixture was stirred vigorously at 25 to 30°C for 6 hours. α〕 ²⁰ _D +93.1゜ (C = 1, methanol) Optical purity 96.2% Example 3 In place of the Arthrobacter lipase in Example 1, Candida cylindrich lipase “Meito MY” (Meito Pharmaceutical 1.05 g of 4(R)-hydroxy-2-cyclopentenone was obtained by the same reaction and post-treatment as in Example 1, except that 200 mg of 4(R)-hydroxy-2-cyclopentenone was used and vigorously stirred for 5 hours at 25-30°C. [α] ²⁰ _D +93.3° (C = 1, methanol) Optical purity 96.4% Example 4 2.0 g of n-propionic acid ester of 4(R)-hydroxy-2-cyclopentenone obtained in Reference Example 2,
200mg of Arthrobacter lipase and 200ml of 0.5M phosphate buffer solution (PH=7) at 25-30℃
Stir vigorously. After the reaction was completed, the mixture was post-treated in the same manner as in Example 1 to obtain 1.01 g of 4(R)-hydroxy-2-cyclopentenone. α] ^26/D +93.1° (C=1, methanol) Optical purity 96.0% Example 5 In place of Arthrobacter lipase in Example 4, Pseudomonas reverse “Amano P” (manufactured by Amano Pharmaceutical Co., Ltd.) 400 mg 1.08 g of 4(R)-hydroxy-2-cyclopentenone was obtained by carrying out the reaction and post-treatment in the same manner as in Example 1, except that the mixture was stirred vigorously at 25-30° C. for hours. α] ²⁰ _D 93.4° (C=1, methanol) Optical purity 96.4% Example 6 2 g of 4(R)-hydroxy-2-cyclopentenone caproic acid ester obtained in Reference Example 3, 300 mg of Arthrobacter lipase and 20 ml of 0.5M phosphate buffer aqueous solution (PH=7) are mixed and stirred vigorously at 25-30°C for 7-8 hours. After the reaction, 4(R)-hydroxy-2-
0.82 g of cyclopentenone was obtained. α] ²⁰ _D +93° (C=1, methanol) Optical purity 96.1% Example 7 β-chloropropionic acid ester of 4(R)-hydroxy-2-cyclopentenone obtained in Reference Example 4 2
g, Arthrobacter lipase 200 mg and
Mix 20 ml of 0.1M phosphate buffer aqueous solution (PH=7) and stir vigorously at 25-30°C for 2-8 hours. After completion of the reaction, post-treatment was carried out in the same manner as in Example 1 to obtain 4(R)-
1.02 g of hydroxy-2-cyclopentenone was obtained. α〕 ²⁰ _D +92.0゜ (C = 1, methanol) Optical purity 95.0% Example 8 In place of the Arthrobacter lipase in Example 7, the lipase of Candeita cylindratsuhi "Meito MY" (manufactured by Meito Pharmaceutical Co., Ltd.) was used. Use 200mg, 25
4(R)-Hydroxy-
1.00 g of 2-cyclopentenone was obtained. [α] ²⁰ _D +91.8゜ (C=1, methanol) Optical purity 94.8%

Claims (1)

[Claims] 1 General formula: (In the formula, R represents a saturated or unsaturated aliphatic hydrocarbon group having 2 to 12 carbon atoms, which may be substituted with halogen. Optically active cyclopentenone esters in which the steric configuration shown by (indicates an atom) is the R configuration are hydrolyzed using an enzyme having esterase activity derived from a microorganism belonging to the genus Arthrobacter, Candida, or Pseudomonas. optically active 4-hydroxy-2- whose steric configuration is R configuration, characterized in that
Method for producing cyclopentenone.