JPH10304893A - Production of optically active compound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To efficiently obtain the subject compound having a high optical purity useful as an intermediate for producing various medicines, by treating a γ-substituted amino-β-hydroxybutyric acid derivative in the presence of a culture solution of a bacterium, a cell (treated substance), etc. SOLUTION: A γ-substituted amino-β-hydroxybutyric acid derivative of the formula (R<1> is an amino-protecting group; R<2> is H, a 1-6C alkyl, a 2-7C alkoxyalkyl) (e.g. 4-benzyloxycarbonylamino-3-hydroxybutanoic acid) is treated in the presence of a culture solution of a bacterium of the genus Brevibacterium such as IFO12, 170, the cell of the bacterium or the treated substance of the bacterium and is converted into one configuration of steric configurations of a hydroxyl group at the β-position to give the objective highly optically active β-substituted amino-β-hydroxybutyric acid derivative useful as an intermediate for producing various medicines.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a process for producing an optically active γ-substituted amino-β-hydroxybutyric acid derivative using a microorganism.

[0002] Optically active γ-substituted amino-β-hydroxybutyric acid derivatives are important as intermediates for the production of various pharmaceuticals.

[0003]

2. Description of the Related Art An optically active .gamma.-
A method for producing a substituted amino-β-hydroxybutyric acid derivative has heretofore been carried out by reacting a racemate with an optically active acid or base to form a salt, which is fractionally recrystallized (JP-A-52-13392).
No. 0) has been known.

[0004]

However, the optical resolution method has problems that an expensive resolving agent is used and the maximum theoretical yield is only 50%.

[0005]

Means for Solving the Problems Accordingly, the present inventors have:
When the compound represented by the general formula (I) is treated with a microorganism, a γ-substituted amino-β having a high optical purity and a high yield is obtained.
-Hydroxybutyric acid derivative was obtained, and the present invention was completed.

That is, the present invention provides a compound represented by the general formula (I)

[0007]

Embedded image [Wherein, R ¹ represents a protecting group for an amino group, and R ² represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or an alkoxyalkyl group having 2 to 7 carbon atoms. Γ
-Substituted amino-β-hydroxybutyric acid derivative is treated in the presence of a culture solution of a microorganism, the microorganism cell or the treated microorganism cell, and the configuration of either one of the hydroxyl group at the β-position is changed to the other. Characterized by being converted to an arrangement of
The present invention relates to a method for producing a γ-substituted amino-β-hydroxybutyric acid derivative having one of the configurations of the hydroxyl group at the β-position.

Further, the present invention provides a γ represented by the general formula (I)
The above process wherein the substituted amino-β-hydroxybutyric acid derivative is a mixture of isomers.

Alternatively, the present invention relates to the above-mentioned production method, wherein the γ-substituted amino-β-hydroxybutyric acid derivative represented by the general formula (I) is a compound having only one of the steric configuration of the β-hydroxyl group.

Further, the present invention relates to the above-mentioned production method, wherein the microorganism is a microorganism having asymmetric oxidation / reduction ability.

The γ-hydroxy group in which the hydroxyl group at the β-position has the R configuration.
The present invention relates to a method for producing a substituted amino-β-hydroxybutyric acid derivative with superiority.

The microorganism is Brevibacterium (Brevibacterium).
bacterium), comprising a γ-substituted amino-β-
The present invention relates to a production method for converting the configuration of a hydroxyl group at the β-position of a hydroxybutyric acid derivative from S to R.

Further, Brevibacterium (Brevibacter)
The present invention relates to the above production method, wherein the microorganism belonging to the genus ium) is IFO12170.

Alternatively, γ in which the hydroxyl group at the β-position is in the S configuration.
A process for obtaining a substituted amino-β-hydroxybutyric acid derivative in an advantageous manner.

The microorganism is Rhodotolur.
a) A microorganism according to the genus of the genus, which relates to the above process for converting the configuration of the hydroxyl group at the β-position of a γ-substituted amino-β-hydroxybutyric acid derivative from R to S.

Furthermore, the present invention relates to the above-mentioned production method, wherein the microorganism belonging to the genus Rhodotlola is IFO0002.

Then, the compound having a hydroxyl group bonded to the asymmetric carbon is treated in the presence of a culture solution of a microorganism of the genus Brevibacterium or Rhodotorula, the microbial cells or a processed product of the microbial cells, The present invention relates to a method for producing a compound having one of the configurations of hydroxyl groups, characterized by converting one of the configurations of hydroxyl groups into the other.

Also, the present invention relates to a method for producing a compound having a hydroxyl group bonded to an asymmetric carbon, wherein the microorganism is a microorganism of the genus Brevibacterium and converts the configuration of the hydroxyl group from S to R.

Furthermore, the present invention relates to the above-mentioned production method, wherein the microorganism of the genus Brevibacterium is IFO12170.

Alternatively, the microorganism is a microorganism of the genus Rhodotorula, and converts the configuration of a hydroxyl group from R to S.
The present invention relates to a method for producing a compound having a hydroxyl group bonded to an asymmetric carbon.

The microorganism of the genus Rhodotorula is an IFO
0002.

The protecting group for the amino group of R ^{1 in} the general formula (I) may be a commonly used protecting group, for example, tertiary butoxycarbonyl group, 2,2,2-trichloroethoxycarbonyl group Alkoxycarbonyl groups such as, benzyloxycarbonyl group, paramethoxybenzyloxycarbonyl group, aralkyloxycarbonyl groups such as paranitrobenzyloxycarbonyl group, acetyl group,
Acyl groups such as methoxyacetyl group, trifluoroacetyl group, chloroacetyl group, pivaloyl group, formyl group, benzoyl group, tertiary butyl group, benzyl group, paranitrobenzyl group, paramethoxybenzyl group, triphenylmethyl group And alkylsulfonyl groups such as a methanesulfonyl group and a trifluoromethanesulfonyl group or a halogenoalkylsulfonyl group, and an arylsulfonyl group such as a benzenesulfonyl group and a toluenesulfonyl group. .

[0023] The production method of the present invention comprises a γ-substituted amino-β-
Recognizing the asymmetric structural portion at the β-position of the hydroxybutyric acid derivative, asymmetric oxidation / reduction of the hydroxyl group of one stereoisomer of the compound shown in Chemical formula 3 occurs.

[0024]

Embedded image

The γ-substituted amino-β-hydroxybutyric acid derivative serving as a substrate may be a mixture of a compound having an R configuration at the β-position and a compound having an S configuration. Or only one of them.

In the case where the configuration at the β-position is a mixture of a compound of R and a compound of S, only a compound having a configuration of either the compound of the β-position or the compound of S is converted, Either the compound of which the configuration at the β-position is R or the compound of S can be preferentially obtained.

When the configuration at the β-position consists of only one of the compound of R and the compound of S, the configuration at the β-position changes from R to S, or the configuration at the β-position changes from S to R. It is possible to obtain a compound which is converted and has a configuration in which the configuration of the β-hydroxyl group is different from that before the treatment.

Further, the asymmetric oxidation and the asymmetric oxidation can be performed by the production method of the present invention.
Examples of the substrate that can be reduced include not only the γ-substituted amino-β-hydroxybutyric acid derivative represented by the general formula (I), but also the same carbon chain as above, with a hydroxyl group at the β-position and a substituted amino group at the γ-position. Any carboxylic acid derivative may be used, and a compound having a substituent at the α-position or a compound further having a substituent at the γ-position is expected to have similar asymmetric recognition, and is expected to undergo asymmetric oxidation and reduction. .

In the present invention, a culture solution of the microorganism, the microorganism cell, or the treated product of the microorganism cell is used. The microorganism may be used properly depending on the configuration of the hydroxyl group of the target compound. For example, when a compound having a hydroxyl group in the R configuration is required, a culture solution of a microorganism belonging to the genus Brevibacterium may be used. When a compound having a hydroxyl group in the S configuration is required, a microorganism belonging to the genus Rhodotorula may be used. A culture solution or the like may be used.

In the present invention, a culture solution of a microorganism belonging to the genus Brevibacterium or Rhodotorula, the microorganism, or a treated product of the microorganism is used.

The culture solution of the microorganism is obtained by culturing the microorganism in an appropriate medium, and contains the microorganism cells grown and their metabolites.

Microbial cells are centrifuged from a culture solution,
Viable cells collected by filtration or the like, or dried cells such as vacuum-dried cells, freeze-dried cells, and cells dried with an organic solvent.

Examples of the treated microorganism cells include extracts of the microorganism cells, ground products of the microorganism cells, ultrasonic treatment of the microorganism cells, and those obtained by known methods such as chromatography and filtration. It is obtained by fractionating and separating components required for the asymmetric oxidation / reduction treatment of the present invention. The components necessary for the processing contained in this fraction may not be completely pure, and may contain other components.

[0034] Alternatively, cells obtained by immobilizing cells on a water-insoluble carrier (for example, acrylamide gel) by a known method may be used.

Microorganisms with IFO numbers are listed in Listof cultures, published by the Institute of Fermentation (IFO).
9th edition (1992) and is available from the IFO.

The production method of the present invention is carried out as follows.

First, a compound represented by the general formula (I) is added to an appropriate culture solution, a culture solution of a microorganism, the microorganism cells or a treated product of the microorganism cells are added, and the reaction solution is cultured with shaking. Do. After completion of the treatment, the target compound can be isolated by using a usual operation such as filtration, liquid separation, and concentration under reduced pressure.

The treatment temperature may be in the range of 10 ° C. to 40 ° C., and preferably around 30 ° C.

The pH of the processing solution may be in the range of 5 to 7.

The treatment time may be in the range of 3 days to 7 days, but is preferably about 5 days.

The concentration of the compound represented by Formula I in the treatment solution is not particularly limited, but is preferably 0.1 to 1.0%.

The obtained compound is useful as an intermediate for the production of various pharmaceuticals and derivatives thereof.

[0043]

Embedded image Oral β- represented by the following structural formula
Methyl carbapenem antibiotics (JP-A-2-22358)
No. 7).

[0044]

Embedded image Hereinafter, the present invention will be described specifically with reference to Examples or Reference Examples, but the present invention is not limited thereto.

[0045]

【Example】

Reference Example 1 4 ml of a normal broth medium (pH 7.0) was dispensed into a test tube and sterilized by heating in an autoclave. A culture of a test tube was inoculated with a seed strain of the genus Brevibacterium (IFO12170), and cultured with shaking at 30 ° C. for 2 nights to obtain a cell culture solution. Example 1 By bacteria of the genus Brevibacterium (IFO12170)
(R) -4-benzyloxycarbonylamino-3-h
Production of droxybutanoic acid 2% glucose, 0.2% peptone, 0.2 enzyme extract
%, Ammonium chloride 1%, dipotassium hydrogen phosphate 0.
25%, potassium dihydrogen phosphate 0.25%, ferrous sulfate
(FeSO ₄ · 7H ₂ O) a liquid medium consisting of 0.009% (pH
7.0) was dispensed into a Sakaguchi flask in a volume of 50 ml and sterilized by heating in an autoclave. 80 mg of 4-benzyloxycarbonylamino-3-hydroxybutanoic acid was aseptically added to this culture solution, and then Brevibacterium (IFO)
12170) was inoculated with 2 ml, and cultured with shaking at 30 ° C. for one week. 5 ml of 1N hydrochloric acid and 100 ml of ethyl acetate were added to the reaction mixture.
Then, the cells were removed by celite filtration. The aqueous layer was further extracted twice with 100 ml of ethyl acetate, the organic layer was dried, and the solvent was distilled off. The residue was subjected to silica gel chromatography to give the target compound as a yellow oil (63 m).
g (optical purity 92% ee) was obtained. In addition, the analysis of the optical isomer was performed using HPLC.

Column used: ULTRON ES-OVM 150 × 4.6 mm (manufactured by Shinwa Chemical Industry Co., Ltd.) Moving layer: 4% methanol (containing 50 mM phosphate buffer)
pH 5.5 Flow rate: 0.8 ml / min, wavelength: 220 nm, column temperature: 40 ° C. NMR (CDCl ₃ ) δ (ppm): 2.5 (2H, dd), 3.1 (1H, dt), 3.3
(1H, m), 3.7 (3H, s), 4.1 (1H, dt), 5.0 (2H, s), 7.4 (5H,
s)

In the same manner, the optically active 4-
Production of benzyloxycarbonylamino-3-hydroxybutanoic acid was performed.

[0048]

[Table 1]

[0049]

According to the present invention, a γ-substituted amino-β-hydroxybutyric acid derivative can be obtained with high purity and a good yield by allowing the reaction to be carried out under mild conditions.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI C12R 1:13) (C12P 13/00 C12R 1: 645)

Claims (10)

[Claims] 1. A compound of the general formula (I) [Wherein, R ¹ represents a protecting group for an amino group, and R ² represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, or an alkoxyalkyl group having 2 to 7 carbon atoms. Γ
A substituted amino-β-hydroxybutyric acid derivative is treated in the presence of a culture solution of a microorganism, the microorganism cells or the treated microorganism cells, and the configuration of either one of the β-position hydroxyl group is changed to the other Characterized by being converted to an arrangement of
Production method for predominantly obtaining a γ-substituted amino-β-hydroxybutyric acid derivative having one of the configurations of the hydroxyl group at the β-position 2. The method according to claim 1, wherein R ² is a hydrogen atom. 3. The method according to claim 1, wherein the γ-substituted amino-β-hydroxybutyric acid derivative represented by the general formula (I) is a mixture of isomers. 4. The production method according to claim 1, wherein the γ-substituted amino-β-hydroxybutyric acid derivative represented by the general formula (I) is a compound having only one of the steric configuration of the hydroxyl group at the β-position. 5. The production method according to claim 1, wherein a compound in which the hydroxyl group at the β-position has the R configuration is predominantly obtained. 6. The microorganism is Brevibacterium (Brev).
6. The method according to claim 5, which is a microorganism belonging to the genus ibacterium. 7. Brevibacterium (Brevibac)
The method according to claim 6, wherein the microorganism belonging to the genus terium is IFO12170. 8. The production method according to claim 1, wherein a compound in which the hydroxyl group at the β-position has the S configuration is predominantly obtained. 9. The method according to claim 8, wherein the microorganism is a microorganism belonging to the genus Rhodotlura. 10. The method according to claim 9, wherein the microorganism belonging to the genus Rhodotolura is IFO0002.