JPH06319572A - Production of halide compound - Google Patents

Abstract

PURPOSE:To simply obtain the compound useful as a raw material for producing medicines. CONSTITUTION:A beta-halogenocarboxylic acid ester having isomers is treated in the presence of a microorganism selected from the microorganisms of the genus Pseudomonas, the genus Bacillus, the Brevibacterium and the Corynebacterium, the culture product of the microorganism, or the extraction fraction of the culture product of the microorganism to obtain the beta- halogenocarboxylic acid or beta-halogenocarboxylic acid ester having a specific steric configuration.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to a synthetic antibacterial drug
No. 231475), a process for producing a compound useful as a starting material.

[0002]

2. Description of the Related Art Equation (2)

[0003]

[Chemical 4] The (1S, 2S) -2-fluorocyclopropane-1-carboxylic acid represented by is, for example, a racemic cis-2-fluorocyclocarboxylic acid obtained by the method described in the literature (Wakayama University, Bulletin of Research 33, 33, 1984). After obtaining propane-1-carboxylic acid, it was obtained by a method in which it was introduced into an amide with an optically active amine and then separated by chromatography, or by a method in which it was introduced into a salt with an optically active base to separate it (Japanese Patent Laid-Open No. H11 (1999) -242242). 2-231
475, JP-A-4-149174).

Further, no microorganisms or enzymes recognizing an asymmetric carbon in which one fluorine atom is substituted have been known so far.

[0005]

Conventionally, in order to obtain a specific stereoisomer, a raw material compound was introduced into a derivative suitable for separation, an optically active salt, or the like for resolution, so that the number of steps is large and the method is industrially used. Was complicated. An object of the present invention is to use a microorganism to obtain a stereoisomer of a compound useful as a raw material for producing a quinolone derivative, in order to easily obtain the desired product without conversion to a derivative or salt for separation. An object of the present invention is to provide a method of optical resolution based on a simple asymmetric hydrolytic method.

[0006]

SUMMARY OF THE INVENTION The present invention is directed to 1,2-cis-2-
Extracting a halogenocyclopropane-1-carboxylic acid ester from a microorganism selected from the group of microorganisms consisting of Pseudomonas, Bacillus, Brevibacterium and Corynebacterium, a culture of the microorganism, or a culture of the microorganism. Characterized by processing in the presence of fractions,
(1S, 2S) -2-halogenocyclopropane-1-carboxylic acid or (1R, 2R) -2-halogenocyclopropane-1-carboxylic acid ester, wherein the ester has 1 carbon atom
To the above-mentioned production method which is an alkyl group, and the above-mentioned production method wherein the halogen atom is a fluorine atom.

Further, the present invention provides the formula (1)

[0008]

[Chemical 5] (Wherein R represents an alkyl group, the fluorine atom and the alkoxycarbonyl group have a cis configuration), and a group of microorganisms consisting of Pseudomonas, Bacillus, Brevibacterium and Corynebacterium A microorganism (2), which is treated in the presence of a microorganism selected from the above, a culture of the microorganism, or an extracted fraction of the culture of the microorganism.

[0009]

[Chemical 6] Or a compound represented by the formula (3)

[0010]

[Chemical 7] (In the formula, R represents an alkyl group.) The present invention relates to a method for producing a compound represented by the formula.

In the compound represented by the formula (1) used in the present invention (hereinafter referred to as the compound (1)), the carboxylic acid moiety may be an alkyl ester, and the alkyl group moiety has 1 to 8 carbon atoms. Groups are preferred. Among them, butyl ester and pentyl ester are particularly preferable. The racemic compound (1) can be easily synthesized by the usual esterification method using the corresponding carboxylic acid compound as a raw material.

The method of the present invention recognizes an asymmetric structural portion of a β-fluorocarboxylic acid compound and causes asymmetric hydrolysis of only one steric carboxylic acid. Substrates that can be asymmetrically hydrolyzed by the method of the present invention include β-chloro or β-bromocarboxylic acids such as β
-Halogenocarboxylic acid compounds, 4-membered ring, 5-membered ring, a further macrocyclic β-halogenocarboxylic acid derivative such as a 6-membered ring, and the same asymmetric recognition is expected in a linear β-halogenocarboxylic acid derivative, Asymmetric hydrolysis is expected.

Therefore, the present invention provides an ester compound in which the carbon atom bonded to the carbon atom to which the carboxyl group is bonded is an asymmetric carbon atom, and the asymmetric carbon atom is substituted by one halogen atom, Treated in the presence of a microorganism selected from the group of microorganisms consisting of Pseudomonas, Bacillus, Brevibacterium and Corynebacterium, a culture of the microorganism, or an extracted fraction of the culture of the microorganism, The present invention relates to a method for converting a carboxylic acid compound having one configuration to a carboxylic acid compound obtained by the method, and further, the present invention relates to a method in which a carbon atom bonded to a carbon atom to which a carboxyl group is bonded is an asymmetric carbon atom. , An ester compound in which the asymmetric carbon atom is substituted by one fluorine atom is a Pseudomonas genus, Bacillus genus, Brevibac After treatment in the presence of a microorganism selected from the group of microorganisms consisting of the genus Lium and Corynebacterium, a culture of the microorganism, or an extracted fraction of the culture of the microorganism, the carboxylic acid compound formed is separated. It also includes a method for obtaining an ester compound and an ester compound obtained by the method.

In the method of the present invention, a microorganism selected from the group consisting of Pseudomonas, Bacillus, Brevibacterium and Corynebacterium can be used. More specifically, Pseudomonas genus, Bacillus genus, Brevibacterium genus and Corynebacterium genus microbial cells selected from the group of microorganisms, a culture of the microbial cells or an extract of the culture of the microorganism. Compound (1) is treated in the presence of the fraction.

The microbial cell culture is obtained by culturing a microbial cell selected from the group consisting of Pseudomonas, Bacillus, Brevibacterium, and Corynebacterium in an appropriate medium. .

The above-mentioned fraction of the culture of microbial cells is the cell or a culture of the cell diluted with water or a suitable buffer, or the filtrate of the culture is ultrafiltered. ,
The components necessary for the treatment are separated by fractionation using a means such as chromatography. It should be noted that the components necessary for the treatment contained in this fraction are not completely pure and may contain some other components.

The method of the present invention is carried out as follows. First, the compound (1) is dissolved or suspended in a suitable solvent or buffer, and then a microbial cell selected from the group consisting of Pseudomonas, Bacillus, Brevibacterium and Corynebacterium, or
A culture of the bacterium or a fraction of the culture may be added, and the mixture may be stirred and treated if necessary to react.

The treatment temperature may be usually in the range of 5 ° C to 60 ° C, but is preferably in the range of 20 ° C to 40 ° C.
The pH of the treatment liquid may be in the range of 4 to 9, but is preferably in the range of 6 to 8.

The reaction time may be 10 hours to 7 days, but usually 20 to 50 hours.

At the start of the reaction, the concentration of the compound (1) in the treatment liquid may be usually 1 to 5% by weight, and particularly preferably 1 to 2%.

The amount of the above-mentioned enzyme, bacterial cells, culture or fraction is not particularly limited, but it may be used in a weight ratio to the compound (1).
0.1-1 times is suitable.

After completion of the treatment, the target compound can be isolated by subjecting the treatment liquid to usual operations such as filtration, liquid separation and concentration under reduced pressure.

[0023]

INDUSTRIAL APPLICABILITY The method of the present invention is a reaction under mild conditions, and side reactions hardly occur. Therefore, a highly pure target compound can be obtained in a high yield and by a simple operation method. It is an industrially excellent method.

[0024]

EXAMPLES The present invention will be described below in greater detail by giving Examples, but the present invention is not limited thereto.

Example 1 Production of cis-2-fluorocyclopropane-1-carboxylic acid butyl ester cis-2-fluorocyclopropane-1-carboxylic acid 4.2 g
Is dissolved in 30 ml of ethylene dichloride, and the butanol 6
g and 0.2 g of concentrated sulfuric acid were added, and the mixture was heated under reflux for 6 hours. Add 200 ml of ethylene dichloride to the reaction mixture to dilute it, and add 2 ml of water to 100 ml.
It was washed once with 100 ml of saturated saline. The organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give the title compound (6.0 g).

¹ H-NMR (CDCl ₃ ) δ: 0.94 (3H, t, J = 7 H
z), 1.01-1.21 (1H, m), 1.32-1.96 (6H, m), 4.15 (2
H, t, J = 7Hz), 4.77 (1H, dm, J = 65Hz)

Example 2 Preparation of cis-2-fluorocyclopropane-1-carboxylic acid ethyl ester cis-2-fluorocyclopropane-1-carboxylic acid 2.1 g
Was dissolved in 20 ml of ethanol, 0.1 g of concentrated sulfuric acid was added, and the mixture was heated under reflux for 4 hours. The reaction mixture was diluted with 100 ml of ethyl acetate and washed twice with 50 ml of water and once with 50 ml of saturated saline. The organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give the title compound (2.5 g).

Reference Example 1 Meat extract 1%, peptone 1
%, And liquid medium (pH 7.2) consisting of 0.5% salt was dispensed into Sakaguchi flask by 100 ml, and sterilized by heating in an autoclave. The medium of each flask was inoculated with an inoculum of the genus Pseudomonas, the genus Bacillus, the genus Brevibacterium or the genus Corynebacterium, and shake-cultured at 30 ° C. for 14 hours. After culturing, cells were obtained by centrifugation, washed with physiological saline, and then freeze-dried to obtain freeze-dried cells.

Example 3 Preparation of (1S, 2S) -2-fluorocyclopropane-1-carboxylic acid and (1R, 2R) -2-fluorocyclopropane-1-carboxylic acid butyl ester cis-2-fluoro Cyclopropane-1-carboxylic acid butyl ester 2.0 g 0.1 M phosphate buffer (pH 7.5) 200 m
l Pseudomonas putida (IAM 150
1.0 g of freeze-dried cells of 6) was added, and the mixture was stirred at 30 ° C for 48 hours (hydrolysis rate was 48%). Chloroform (100 ml) was added to the reaction solution, and the mixture was stirred for 30 minutes, and the cells were removed by filtration through Celite. Further, the aqueous layer was extracted twice with 100 ml of chloroform. The organic layer was washed twice with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure (1R, 2
830 mg (41.5%, 98% ee) of R) -2-fluorocyclopropane-1-carboxylic acid butyl ester was obtained.

¹ H-NMR (CDCl ₃ ) δ: 0.94 (3H, t, J = 7 H
z), 1.01-1.21 (1H, m), 1.32-1.96 (6H, m), 4.15 (2
H, t, J = 6 Hz), 4.77 (1H, dm, J = 65 Hz)

The aqueous layer after extraction with chloroform was adjusted to pH 2
Then, the product was extracted 3 times with 100 ml of chloroform and treated in the same manner as above to give 510 mg (39.2%, 92% ee) of (1S, 2S) -2-fluorocyclopropane-1-carboxylic acid. It was

¹ H-NMR (CDCl ₃ ) δ: 1.01-1.42 (1H, m),
1.56-1.98 (2H, m), 4.76 (1H, dm, J = 66 Hz), 11.32
(1H, s)

The analysis of optical isomers was carried out by using gas chromatography.・ Column used: CP cyclodex β-236M (GL Science)

Example 4 Production of (1S, 2S) -2-fluorocyclopropane-1-carboxylic acid and (1R, 2R) -2-fluorocyclopropane-1-carboxylic acid ethyl ester cis-2-fluoro Cyclopropane-1-carboxylic acid ethyl ester 1.0 g 0.1 M phosphate buffer (pH 7.5) 100 m
l Pseudomonas putida (IAM 150
1.0 g of freeze-dried cells of 6) was added, and the mixture was stirred at 30 ° C for 48 hours (hydrolysis rate was 52%). Chloroform (40 ml) was added to the reaction solution, and the mixture was stirred for 30 minutes, and the cells were removed by Celite filtration. Further, the aqueous layer was extracted twice with 40 ml of chloroform. The organic layer was washed twice with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give (1R, 2R) -2-
Fluorocyclopropane-1-carboxylic acid ethyl ester
405 mg (40.5%, 96% ee) was obtained.

The aqueous layer after extraction with chloroform was adjusted to pH 2
Was extracted with 40 ml of chloroform three times, treated in the same manner as above, and the solvent was distilled off to give (1S, 2S) -2.
-Fluorocyclopropane-1-carboxylic acid 300 mg (38.0
%, 90% ee) was obtained.

Example 5-8 Preparation of (1S, 2S) -2-fluorocyclopropane-1-carboxylic acid cis-2-fluorocyclopropane-1-carboxylic acid butyl ester (500 mg) was mixed with 0.1 M phosphoric acid. Buffer solution (pH 7.5) 5
Frozen cells of the strains shown in Table 1
00 mg was added and it stirred at 30 degreeC for 48 hours. Thereafter, the same operation as in Example 4 was performed to obtain the target product. The results are shown in Table 1.

[0037]

[Table 1]

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Office reference number FI technical display location // (C12P 7/40 C12R 1:40) (C12P 7/40 C12R 1:07) (C12P 7 / 40 C12R 1:13) (C12P 7/40 C12R 1:15) (C12P 7/40 C12R 1:11) (C12P 7/62 C12R 1:38)

Claims (4)

[Claims] 1. A 1,2-cis-2-halogenocyclopropane-1
-The presence of a carboxylic acid ester in a microorganism selected from the group of microorganisms consisting of Pseudomonas, Bacillus, Brevibacterium and Corynebacterium, a culture of the microorganism, or an extracted fraction of the culture of the microorganism. Process for producing (1S, 2S) -2-halogenocyclopropane-1-carboxylic acid or (1R, 2R) -2-halogenocyclopropane-1-carboxylic acid ester, characterized by being treated below 2. The method according to claim 1, wherein the ester is an alkyl group having 1 to 6 carbon atoms. 3. The method according to claim 1, wherein the halogen atom is a fluorine atom. 4. Formula (1): (Wherein R represents an alkyl group, the fluorine atom and the alkoxycarbonyl group have a cis configuration), and a group of microorganisms consisting of Pseudomonas, Bacillus, Brevibacterium and Corynebacterium Wherein the treatment is carried out in the presence of a microorganism selected from the group consisting of, a culture of the microorganism, and an extracted fraction of the culture of the microorganism. Or a compound represented by the formula (3): (In the formula, R represents an alkyl group.) A method for producing a compound represented by the formula.