JPS62257396A - Production of optically active 4-hydroxy-2-cyclopentenone - Google Patents

Abstract

PURPOSE:To produce the titled substance useful as a raw material for prostaglandin, by hydrolyzing an optically active cyclopentenone ester with esterase. CONSTITUTION:An optically active cyclopentenone ester is obtained by converting an optically active 4-hydroxy-2-cyclopentenone into a sulfonic ester, treating the sulfonic ester with an acetate replaced with a halogen atom and carrying out conversion to give an optically active cyclopentenone ester shown by the formula. When a compound in R coordination is desired as the aimed compound, 4-hydroxy-2-cyclopentenone is S coordination is used as a raw material. The prepared optically active cyclopentenone is hydrolyzed with esterase derived from a bacterium or animal or plant in a buffer solution by stirring to give the aimed compound.

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to a method for producing optically active 4-hydroxy-2-cyclopentenone.

<Prior art> Optically active 4-hydroxy-2-cyclopentenone is useful as a raw material for pharmaceuticals, especially prostaglandins that have various pharmacological effects such as anti-ulcer and thrombolytic effects, and the purpose of 1ξ is known. ing.

Conventionally, as a method for producing such optically active 4-hydroxy-2-cyclopentenone, for example, acetate ester of 4-hydroxy-2-cyclopentenone having an R coordination is prepared using wheat germ lipase enzyme, and 48 A method (Japanese Patent Publication No. 55-84677) is known in which hydrolysis takes a long time.

However, this method is extremely inefficient because it requires a very long reaction time, and furthermore, if the reaction is left under hydrolysis conditions for a long time, racemization will proceed simultaneously and the optical purity will decrease. was clarified by the research conducted by the present inventors, but since high optical purity is required when optically active 4-hydroxy-2-cyclopentenone is used for pharmaceutical purposes, this method is particularly applicable to pharmaceutical applications. It is unsatisfactory as a manufacturing method for use as a raw material.

<Problems to be Solved by the Invention> Based on the above, the present inventors have solved the problems of the above-mentioned prior art and have achieved optically active production in a short time, easily, and without racemization. As a result of a study to produce optically active 4-hydroxy-2-cyclopentenone by hydrolyzing cyclopentenone esters, halogenoacetate was used instead of acetate as the raw material optically active cyclopentenone ester. The inventors have discovered that the above object can be achieved by doing the following, leading to the present invention.

<Means for Solving the Problems> That is, the present invention provides a compound represented by the general formula (1) (wherein, R represents a methyl group substituted with a halogen atom. The square symbol represents an asymmetric carbon atom). The present invention provides a method for producing optically active 4-hydroxy-2-cyclopentenone, which comprises hydrolyzing optically active cyclopentenone esters using esterase.

In the present invention, optically active cyclopentenone esters as raw materials are obtained by, for example, converting optically active 4-hydroxy-2-cyclopentenone into a sulfonic acid ester, which is then treated with an acetate substituted with a halogen atom, and then inverted. can be obtained by preparing an optically active cyclopentenone ester represented by the above general formula (1).

In the method of the above formula, optically active 4-hydroxy-2-cyclopentenone as a raw material has a structure. -2-cyclopentenone is used as a raw material.

This means that, for example, when a compound with R coordination is desired as optically active 4-hydroxy-2-cyclopentenone,
This is extremely important for making effective use of unnecessary S-coordination.

Examples of such optically active cyclopentenone esters include chloroacetate, dichloroacetate, trichloroacetate, and bromoacetate.

Hydrolysis of optically active cyclopentenone esters is
Hydrolysis is carried out using esterases produced by microorganisms or esterases derived from animals and plants.

In the reaction (B), since the optically active cyclopentenone esters used as raw materials are extremely easily hydrolyzed, the esterase can be used without any particular selection.

Such esterases refer to esterases in a broad sense, including lipases.

Here, specific examples of the above-mentioned microorganisms include Enterobacter genus, Arthrobacter genus, Brevibacterium genus, Pseudomonas genus, alkaline earth metals, Micrococcus genus, Chromobacterium genus, Microbacterium genus, Corynebacterium spp. genus Bacillus, genus Lactobacillus, genus Trichoderma, genus Campita, genus Salacharomyces, genus Rhodotorula, genus Cryptococcus, genus Torulopsis, genus Pichia, genus Penicillium, genus Asperbolus, genus Rhizopus, genus Mucor, genus Aureobasidium. genus,
Examples include microorganisms belonging to the genus Actinomyces, genus Nocardia, genus Streptomyces, genus Hansenula, and genus Achromobacter.

For culturing the above-mentioned microorganisms, a culture solution is usually obtained by carrying out liquid culture according to a conventional method.

Furthermore, some of these microbial-derived esterases are commercially available and can be easily obtained. Specific examples of commercially available esterases include the following.

Lipase from the genus Pseudomonas (manufactured by Amano Pharmaceutical), lipase from the genus Aspergillus [Lipase AP (manufactured by Amano Pharmaceutical)], lipase from the genus Mucor (M-AP) (manufactured by Amano Pharmaceutical), lipase from Candida cylindraffeus [Lipase MY (manufactured by Meito Sangyo)] )].

Alkaline earth metal lipase [Lipase PL (manufactured by Meito Sangyo)],] Achromobacter Nori Perse [Lipase AL
(manufactured by Meito Sangyo)],] lipase of Arthrobacter (manufactured by Shin Nippon Kagaku), lipase of Chromobacterium (manufactured by Toyo Jozo), lipase of Rhizopus delemer [Talipase (manufactured by Tanabe Seiyaku)], lipase of Rhizopus sp. lipase [Lipase Saiken (Osaka Bacteria Research Institute)].

Further, animal/plant esterases can also be used, and specific examples of these esterases include the following.

Steapsin, pancreatin, pig liver esterase, What Germ esterase.

As the esterase used in this reaction, enzymes obtained from animals, plants, and living organisms are used, and the usage forms include purified enzyme, crude enzyme, enzyme-containing preparation, microbial culture solution, culture, bacterial cells, It can be used as needed in various forms such as cultured coins and processed products, and m5 and microorganisms can also be used in combination. Alternatively, it can also be used as an immobilized # element immobilized on a resin or the like, or as an immobilized bacterial cell.

The hydrolysis reaction of optically active cyclopentenone esters is usually carried out by vigorously stirring a mixture of the raw material dL-cyclopentenone ester and the above-mentioned enzyme or microorganism in a buffer solution.

As the buffer, commonly used buffers of inorganic acid salts such as sodium phosphate and potassium phosphate, buffers of organic acid salts such as sodium acetate and sodium citrate, etc., are used, and the pH thereof is usually pH 5-5. 8 is preferred. In addition, its concentration is usually 0.05 to 2M, preferably 0.05M.
The range is M.

The reaction temperature is usually 10-60''C.

The reaction time is within 0.5 to 10 hours, often within 6 hours.

After the hydrolysis reaction, the product can be easily separated from the reaction solution by using conventional separation methods such as extraction, separation, concentration, distillation, and chromatography.

<Effects of the Invention> Thus, according to the method of the present invention, optically active 4-hydroxy-2-cyclopentenone can be easily obtained in a short time without racemization and therefore without impairing optical purity. Can be done.

Furthermore, according to the method of the present invention, the optically active cyclopentenone ester represented by the general formula (1) is easily hydrolyzed by esterase as described above, so that the optically active 4-hydroxyl-2-cyclo After converting pentenone into a sulfonic acid ester, it is treated with a halogen-substituted acetate to obtain an optically active cyclopentenone ester represented by the general formula (1), which is then hydrated using the method of the present invention. Through a series of decomposition operations, it can also be used as a stereoinversion reaction to obtain 4-hydroxy-2-cyclopentenone, which has an optical configuration opposite to that of the starting material, optically active 4-hydroxy-2-cyclopentenone. available and extremely useful.

<Example> The present invention will be explained below with reference to Examples.

Reference Example 1 4 (S)-Hydroxy-2-cyclopentenone (S)-hydroxy-2-cyclopentenone (
Optical purity 97%) 29.4f, dichloromethane 150-
and 86.7 g of pyridine, and 87.81 g of methanesulfonyl chloride was added over 8 hours at -10°C.

After incubating at the same temperature for 1 hour, the reaction solution was mixed with water, 2% sodium chloride water,
Wash sequentially with water. The organic layer is dried over magnesium sulfate and then concentrated. The concentrated residue was dissolved in toluene-ethyl acetate-6
Perform silica gel column chromatography using a mixed solution of =8.

4 Methanesulfonic acid ester of (S)-hydroxy-2-cyclopentenone 49.21 is obtained.

no LA 855 (It will crystallize if left alone.)
16 g of the obtained methanesulfonic acid ester of 4(S)-hydroxy-2-cyclopentenone
Add 27.81 L/sodium acetate and hexamethylphosphorotria 0- and react at 80-40°C for 6 hours.

After the reaction is completed, the reaction solution is added to water, and extracted with 160% of toluene.

The organic layer was sequentially washed with 2% sodium hydrogen chloride, water, 2% hydrochloric acid, and water, dried over magnesium sulfate, and concentrated under reduced pressure using a mixture of ethyl acetate and toluene (1=6). chromatographic purification to obtain 17.7'rfl of dichloroacetate of 4(2)-hydroxy-2-cyclopentenone.

iz ] no + 118.9・ (c = 1, C
HC'AI)m-p 78.5-74.6°C Reference Example 2 Sodium monochloroacetate 21.1f was added to 4(S)-hydroxy-2-cyclopentenone methanesulfonic acid ester 161 obtained in Reference Example 1. and hexamethylphosphorotria electrolyte 〇- are added, and the mixture is allowed to react at 80 to 40 C for 7 hours.

After the reaction, post-treatment and purification were carried out in the same manner as in Reference Example 1 to obtain 4(9
)-Hydroxy-2-cyclopentenone monochloroacetate 14. ' get lfl.

a] D+ 129-40 (c = 1, CHC
ta) m*p* 75-76,5°C Reference Example 8 Sodium bromoacetate 28.981 was added to 15.87F methanesulfonic acid ester of 4(S)-hydroxy-2-cyclopentenone obtained in Reference Example 1. and hexamethylphosphorotria [4(2)-hydroxy-2]
- Bromoacetic acid ester of cyclopentenone 18.78F
get.

a]no + 1074・(c=1.methanol)m
, p60-61°C Example 1 4 CR)-Hydroxy-2-! obtained in Reference Example 1 /cyclobentenone dichloroacetate 2.0911 Aspergillus lipase [Lipase At' (manufactured by Ohno Pharmaceutical Co., Ltd.)
1200v and 0.8M phosphate buffer aqueous solution (p
H7) 100 d were mixed and stirred vigorously at 26-80''C for 8 hours. After the reaction was completed, Glauber's salt was added to the reaction solution, and extracted with methyl isobutyl ketone.

The extract was concentrated and distilled using a microdistillation device to obtain 0.87 g of 4(9)-hydroxy-2-cyclopentenone.

αID+91.9・(c=1.methanol) Optical purity
94.9% The optical purity was measured by NMR after converting it into an ester of (→-α-methoxyalpha (trifluoromethyl)-phenylacetic acid).

Example 2 Wheat germ lipase (manufactured by Sigma) 100q was used instead of Aspergillus lipase at 17125-80°C
The reaction was carried out in the same manner as in Example 1, except that the reaction mixture was vigorously stirred for 6 hours, and after treatment, 4(9)-hydroxy-2-cyclopentenone 0,85F was obtained.

α]D+91.8・(c=1.methanol) Optical purity 9
4.1% Examples 8 to 6 Reactions and post-treatments were carried out in accordance with Example 1, except that the type of lipase, the amount used, and the reaction time were as shown in Table 1.

The results are shown in Table-1.

Example 7 Monochloroacetic acid ester 1, r6y of 4 (R)-hydroxy-2-cyclopentenone obtained in Reference Example 2, Pseudomonas lipase "Amano JP (manufactured by Ohno Pharmaceutical Co., Ltd.) 80
■ and 0.8 M phosphate buffer aqueous solution (pH 7)
Mix 1oo- and stir vigorously for 4 hours at 25-80°C. After completion of the reaction, post-treatment and purification were performed according to Example 1 to obtain 4Q? )-Hydroxy-2-cyclopentenone 0.8
Get 49.

a') D+ 91-6 @ (c-1 + methanol) Optical purity 94.6% / 1 gM Examples 8 to 10 Performed except for the type of lipase, the amount used, and the reaction time as shown in Table 2. The reaction and post-treatment were carried out according to Example 7. The results are shown in Table-2.

Example 11 Monobromoacetate of 4(2)-hydroxy-2-cyclopentenone obtained in Reference Example 8 2.21. Arthrobacter lipase (manufactured by Shin Nippon Kaisha) 60Iv and 0.
Mix 100-8M phosphate buffer aqueous solution (pH 7) and stir vigorously at 25-80°C for 8 hours. After completion of the reaction, post-treatment and purification were performed according to Example 1 to obtain 4(9).
-Hydroxy-2-cyclopentenone 0.861 is obtained.

a]D +91.8@(C-1, methanol) Optical purity 9468% Examples 12-1B Follow the same procedure as Example 11 except that the type of lipase, the amount used, and the reaction time are as shown in Table-8. Reaction and post-treatment were performed. The results are shown in Table-8.

Claims (2)

[Claims] (1) General formula ▲ Numerical formulas, chemical formulas, tables, etc. are available ▼ (In the formula, R represents a methyl group substituted with a halogen atom. The * mark represents an asymmetric carbon atom.) Pentenone esters,
A method for producing optically active 4-hydroxy-2-cyclopentenone, characterized by hydrolysis using esterase (2) The steric configuration of the optically active cyclopentenone ester is the R configuration, and the steric configuration of the optically active 4-hydroxy-2-cyclopentenone is the R configuration.Claim 1 Manufacturing method described in section