JP2003000295A - Method for producing optically active 2-hydroxymethyl-3- arylpropionic acid and antipode ester thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for readily producing an optically active 2- hydroxymethyl-3-arylpropionic acid, and to obtain an antipode ester thereof. SOLUTION: The optically active 2-hydroxymethyl-3-arylpropionic acid represented by formula (2) and the antipode ester thereof are produced by asymmetrically hydrolyzing a 2-hydroxymethyl-3-arylpropionic acid ester represented by formula (1) by using an enzyme.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing optically active 2-hydroxymethyl-3-arylpropionic acid and its antipodal ester. Optically active 2-hydroxymethyl-3-arylpropionic acid, especially optically active 2
-Hydroxymethyl-3-phenylpropionic acid is very useful as a raw material for optically active 2-acetylthiomethyl-3-phenylpropionic acid, which is used as an intermediate for the production of pharmaceuticals such as enkephalinase inhibitors and ACE inhibitors. It is a compound.

[0002]

2. Description of the Related Art As a method for producing optically active 2-hydroxymethyl-3-phenylpropionic acid, which is one of optically active 2-hydroxymethyl-3-arylpropionic acids, various methods have hitherto been used. It has been known. (A) 2-benzyl-1,3-propanediol is asymmetrically esterified with lipase in the presence of vinyl acetate, or 2- (acetoxymethyl) -3-phenylpropyl acetate is asymmetrically hydrolyzed with lipase. A method for obtaining optically active 2-hydroxymethyl-3-phenylpropionic acid [WO9729086, Tetrahedro] by oxidizing optically active 2-benzyl-3-hydroxypropyl acetate obtained by the above method with chromic acid and then performing deacetylation
n Lett. 31 , 1601 (1990)]. (B) (4S) -4- (1-Methylethyl) -2-oxazolidinone was reacted with n-butyllithium and 3-phenylpropionyl chloride to give phenylpropionylamide, and then reacted with lithium diisopropylamide. A method in which the produced enolate is reacted with benzyl bromomethyl ether for asymmetric alkylation, and the resulting compound is further subjected to two steps to obtain optically active 2-hydroxymethyl-3-phenylpropionic acid [Chem
Lett. 505 (1990)]. (C) (R) -2-benzyl-3- (formyloxy) obtained by subjecting (S) -2-isocyano-3-phenylpropyl formate obtained in 3 steps from L-phenylalanine to a high thermal rearrangement reaction. Optical activity of propionitrile by hydrolysis 2-
Method for obtaining hydroxymethyl-3-phenylpropionic acid [Chem Ber. 123, 635 (199
0)].

However, the above method (a) is difficult to industrialize, such as using toxic chromic acid when oxidizing the alcohol obtained by the enzymatic reaction into a carboxylic acid. Also,
The method (b) is not an industrial method because it has many steps and uses expensive n-butyllithium or lithium diisopropylamide and requires a low temperature reaction. In addition, the method (c) above is performed by thermal rearrangement reaction at 500
It requires a high temperature of ℃ or more, and cannot be an industrial manufacturing method. As described above, all the conventionally known methods have problems as industrial manufacturing methods.

[0004]

Therefore, the present inventors have found that
As a result of diligent studies to develop a method capable of producing optically active 2-hydroxymethyl-3-arylpropionic acid from 2-hydroxymethyl-3-arylpropionic acid ester in one step with good optical purity, 2-hydroxymethyl-3 It was found that an optically active 2-hydroxymethyl-3-arylpropionic acid and its enantiomer ester can be easily produced with a high optical purity by asymmetrically hydrolyzing an arylpropionic acid ester with an enzyme, and the present invention provides I arrived.

[0005]

That is, the present invention provides a compound represented by the general formula (1);

[0006]

[Chemical 3] (In the formula, Ar is an aryl group which may have a substituent,
R represents an alkyl group, an aralkyl group or an aryl group, and * represents an asymmetric carbon atom. ), A 2-hydroxymethyl-3-arylpropionic acid ester is reacted with an enzyme having an asymmetric hydrolysis ability, a general formula (2);

[0007]

[Chemical 4] (In the formula, Ar and * have the same meanings as described above.)
The present invention provides a method for producing the optically active 2-hydroxymethyl-3-arylpropionic acid and its enantiomer ester.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION In the 2-hydroxymethyl-3-arylpropionic acid ester represented by the general formula (1), Ar represents an aryl group such as a phenyl group and a naphthyl group, and may have a substituent. Good. Examples of the substituent include an alkyl group, a substituted alkyl group, an alkoxy group, a substituted alkoxy group, a hydroxyl group, an amino group, a nitro group and a halogen atom. Ar is preferably a phenyl group which may have the above-mentioned substituent, more preferably a phenyl group.

In the 2-hydroxymethyl-3-arylpropionic acid ester represented by the general formula (1), R is
Examples of the alkyl group represented by are lower alkyl groups having 1 to 8 carbon atoms such as a methyl group, an ethyl group, a propyl group, and a butyl group, and examples of the aralkyl group include a benzyl group and an arylethyl group, and an aryl group. Examples thereof include a phenyl group. Preferably a methyl group,
Number of carbon atoms such as ethyl, propyl and butyl
8 lower alkyl groups, particularly preferably a methyl group.

The 2-hydroxymethyl-3-arylpropionic acid ester represented by the general formula (1) has two kinds of optically active substances having an asymmetric carbon atom represented by * as an asymmetric center. The 2-hydroxymethyl-3-arylpropionic acid ester (1) used in the method of the present invention may be a racemate containing an equal amount of each of these optically active substances, or may contain one of the optically active substances in excess. It may be an optically active compound.

The 2-hydroxymethyl-3-arylpropionic acid ester (1) can be synthesized by several known methods. For example, β-propiolactone is subjected to ring opening reaction with methanol in the presence of triethylamine to give 3
-Hydroxypropionic acid methyl ester was added, and then lithium diisopropylamide and benzyl bromide were used to
-Benzylation to give 2-hydroxymethyl-3-phenylpropionic acid methyl ester (J. Me
d. Chem. 1993, 36, 4015) and the like.

The enzyme capable of asymmetrically hydrolyzing 2-hydroxymethyl-3-arylpropionic acid ester (1) may be an enzyme of microbial origin or an enzyme of animal origin. The enzyme may be of plant origin.

[0013] Examples of the enzymes of microbial origin, for example Achromobacter (Achromobacter) genus Aspergillus (Aspergillus) genus Bacillus (B
genus acillus , Candida
Genus, Chromobacterium
um) genus, Humicola (Humicola) genus, Mucor (Mucor) genus Penicillium (Penicill
ium genus, Pseudomona
s) genus and Rhizopus (Rhizopus) enzyme and the microorganism origin or recombinant microorganism transformed by the gene is introduced with these microorganisms are like enzyme produced, preferably, chromo Bacteria ( Chromobacterium )
It is an enzyme originating from a microorganism such as the genus Pseudomonas .

Any of these microorganisms can be used in a conventional manner, for example, 20 to 40 by inoculating the sterilized liquid medium with the microorganism.
Liquid culture can be easily carried out by a method such as reciprocal shaking culture at ℃. In addition, solid culture may be carried out if necessary.

Examples of the enzyme of animal origin include enzymes derived from the internal organs of pigs and cows, preferably the enzymes derived from the pancreas of pigs and cows. Examples of plant-derived enzymes include enzymes originating from wheat germ.

These enzymes can be selected from commercial products and used. Examples of commercially available enzymes include lipase AL (derived from Achromobacter sp. , Manufactured by Meito Sangyo Co., Ltd.), Patalase A ( Asper
derived from gills niger , NOVO), lipase SP539 (derived from Bacillus sp. , NO)
VO), Subtilisin A ( Bacillus li)
cheniformis- derived, NOVO), lipase OF ( Candida cylindracea- derived, Meito Sangyo), lipase SP398 ( Candi )
da sp. Origin, NOVO), Lipase L-05
0 ( Chromobacterium viscosu
m origin, manufactured by Biocatalysts), lipase L-053 ( Humicola lanuginosa)
Origin, manufactured by Biocatalysts), Lipase L
-166P (from Mucor javanicus ,
Manufactured by Biocatalysts), Lipozyme
10000L (from Mucormeihei , NOV
O company), lipase SP388 ( Mucor mieh
derived from ei , manufactured by NOVO), lipase L-055P
( Derived from Penicillum cyclopium ,
Biocatalysts), lipase PS ( Ps
derived from Eudomonas cepacia , manufactured by Amano Pharmaceutical Co., Ltd., Lipase L-056 ( Pseudomonas)
Fluorescens origin, Biocatalys
ts), Lipase D ( Rhizopus dele
derived from mar , manufactured by Amano Pharmaceutical Co., Ltd.), lipase F-AP15
( Rhizopus javanicus origin, manufactured by Amano Pharmaceutical Co., Ltd.), lipase (porcine pancreas origin, manufactured by Toyo Brewing Co., Ltd.), chymotrypsin (bovine pancreas origin, manufactured by Nacalai Tesque, Inc.) and the like.

The purity or form of these enzymes is not particularly limited, and various kinds of purified enzymes, crude enzymes, enzyme-containing substances, microbial culture liquids, cultures, cells, culture liquids and processed products thereof can be used. Can be used in the form of. Here, the treated product refers to, for example, freeze-dried microbial cells, acetone-dried microbial cells, bacterial rubs, autolyzed microbial cells, ultrasonically crushed microbial cells, microbial cell extract or alkali-treated product. . Furthermore, various purities or forms of enzymes as described above are added to inorganic carriers such as silica gel and ceramics,
It may be immobilized by a known method such as an adsorption method on cellulose, an ion exchange resin, etc., a polyacrylamide method, a sulfur-containing polysaccharide gel method (for example, a carrageenan gel method), an alginic acid gel method, an agar gel method and the like.

Such an enzyme is appropriately selected according to the desired optically active 2-hydroxymethyl-3-arylpropionic acid (2).

The asymmetric hydrolysis is carried out, for example, by mixing water, 2-hydroxymethyl-3-arylpropionic acid ester and the above enzyme.

The amount of the enzyme used is appropriately selected so that the reaction time is not delayed and the selectivity is not lowered. For example, when a commercially available product is used, the amount used is 2-hydroxymethyl-3-
The arylpropionic ester (1) is usually 0.001 to 2.0 times by weight, preferably 0.01 to 1.
It is 0 times the weight.

The water used for the asymmetric hydrolysis may be a buffered aqueous solution. Examples of the buffer aqueous solution include a buffer aqueous solution of an inorganic acid salt such as an aqueous solution of an alkali metal salt of phosphate such as an aqueous solution of sodium phosphate and an aqueous solution of potassium phosphate, an organic acid salt such as an aqueous solution of sodium acetate and an aqueous solution of potassium acetate. Buffered aqueous solution and the like. The amount of such water used is usually 0.5 mol times or more with respect to the 2-hydroxymethyl-3-arylpropionic acid ester (1), and in some cases, it is used as a solvent amount, and usually 100 times by weight or less. is there.

The pH of the reaction system is appropriately selected so that the asymmetric hydrolysis by the enzyme proceeds with good selectivity and is not particularly limited, but it is usually in the range of pH 4 to 10. The reaction temperature is
If it is too high, the stability of the enzyme tends to decrease, and if it is too low, the reaction rate tends to decrease.
The temperature is 65 ° C, and preferably in the range of 20 to 50 ° C.

In the present invention, one optically active substance of 2-hydroxymethyl-3-arylpropionic acid ester (1) maintains the configuration around the asymmetric carbon atom represented by * by asymmetric hydrolysis. As it is, it is preferentially hydrolyzed to give the desired optically active 2-hydroxymethyl-
3-Arylpropionic acid (2) is produced, and its antipodal ester remains without being hydrolyzed.

After the reaction solution after the reaction is adjusted to be alkaline, an organic solvent is added, and the mixture is sufficiently stirred and then liquid-separated into an aqueous layer and an organic layer to obtain an optically active 2-hydroxymethyl-3-hydrate as an aqueous layer. An aqueous solution of arylpropionic acid (2) can be used as an organic layer to obtain a solution of enantiomer ester. Examples of the organic solvent that can be used at this time include ethers such as t-butylmethyl ether and isopropyl ether, hydrocarbons such as toluene, hexane, cyclohexane, and heptane, dichloromethane, dichloroethane, chloroform, chlorobenzene, orthodichlorobenzene, and the like. Examples thereof include halogenated hydrocarbons, esters such as ethyl acetate, methyl acetate and butyl acetate.

Then, water is distilled off from the obtained aqueous solution of optically active 2-hydroxymethyl-3-arylpropionic acid (2), or the aqueous solution is extracted with an organic solvent under acidic conditions, and the solvent is distilled off. By carrying out, the target optically active 2-hydroxymethyl-3-arylpropionic acid (2) can be obtained.

The enantiomer ester 2-hydroxymethyl-3-arylpropionic acid ester (1) which has not been hydrolyzed is contained in the organic layer after liquid separation. Can be easily taken out from the organic layer. The 2-hydroxymethyl-3-arylpropionic acid ester (1) thus obtained is hydrolyzed by a conventional method, neutralized with an acid, extracted with an organic solvent and evaporated to remove an optically active 2-hydroxy compound. It is possible to obtain methyl-3-arylpropionic acid (2). As the hydrolysis method, hydrolysis can be carried out under usual conditions, and examples thereof include a method of dissolving in methanol and then adding an aqueous sodium hydroxide solution.

The optically active 2-hydroxymethyl-3-arylpropionic acid (2) thus obtained can be further purified by recrystallization, column chromatographic treatment or the like. Moreover, the optical purity can be increased by recrystallization.

[0028]

EXAMPLES The present invention will be described in more detail with reference to examples below, but the present invention is not limited to these examples.

(Examples 1 to 7) 10 mg of racemic methyl 2-hydroxymethyl-3-phenylpropionate was suspended in 1 ml of 500 mM phosphate buffer (pH 7), and 5 mg of various enzymes shown in Table 1 were added. And stirred at 30 ° C. for 19 hours. After completion of the reaction, 2N hydrochloric acid was added to the reaction solution to make it acidic, and the mixture was extracted with t-butyl methyl ether. After timely concentration of the extract, high performance liquid chromatography (column:
Finepak SIL C18T made by JASCO Corporation
-5, eluent: water (0.1% phosphoric acid) / acetonitrile = 7/3, flow rate: 1.0 ml / min, detection: 220n
m, column temperature: 35 ° C., elution time: 2-hydroxymethyl-3-phenylpropionic acid 6.6 min, 2-hydroxymethyl-3-phenylpropionic acid methyl 1
2.7 min), 2-hydroxymethyl-3
-The conversion rate to phenylpropionic acid was determined. Further, the produced 2-hydroxymethyl-3-phenylpropionic acid was purified by thin layer chromatography, and high performance liquid chromatography (column: Chiralcel OD manufactured by Daicel Chemical Industries Ltd., eluent: hexane / isopropanol / trifluoroacetic acid = 95/5 / 0.0
5, flow rate: 1.0 ml / min, detection: 215 nm, column temperature: room temperature, elution time: S-form 15.6 min, R-form 17.6 min) to measure the optical purity. The results are shown in Table 1.

[0030]

[Table 1] (Examples 8 to 18) Racemic 2-hydroxymethyl-
Methyl 3-phenylpropionate 10 mg 500 mM
The suspension was suspended in 1 ml of a phosphate buffer (pH 7), 10 mg of various enzymes shown in Table 2 were added, and the mixture was stirred at 30 ° C. for 48 hours. After the reaction, extraction and analysis are performed in the same manner as in Examples 1 to 7,
The conversion rate to 2-hydroxymethyl-3-phenylpropionic acid and the optical purity of the produced 2-hydroxymethyl-3-phenylpropionic acid were measured. The results are shown in Table 2.

[0031]

[Table 2] Example 19 Racemic 2-hydroxymethyl-3-
10 mg of methyl phenylpropionate was suspended in 1 ml of 500 mM phosphate buffer (pH 7), 5 mg of the enzyme shown in Table 3 was added, and the mixture was stirred at 30 ° C. for 5 hours. After the reaction, extraction and analysis were performed in the same manner as in Examples 1 to 7 to measure the conversion rate to 2-hydroxymethyl-3-phenylpropionic acid and the optical purity of the produced 2-hydroxymethyl-3-phenylpropionic acid. . The results are shown in Table 3.

[0032]

[Table 3] Example 20 Racemic 2-hydroxymethyl-3-
Lipase L- was prepared by suspending 1.0 g of methyl phenylpropionate in 100 ml of 500 mM phosphate buffer (pH 7).
050 (manufactured by Biocatalysts) was added, and the mixture was stirred at 30 ° C. for 5 hours. After the reaction was completed, the enzyme was removed by filtration. The reaction mixture was adjusted to pH 9 by adding a 1N aqueous sodium hydroxide solution under ice cooling, 100 ml of t-butyl methyl ether was added, and the mixture was stirred and the organic layer was removed. The resulting aqueous layer was adjusted to pH 3 by adding 2N hydrochloric acid under ice cooling, and t-butyl methyl ether 10 was added.
Extracted twice with 0 ml. The organic layers were combined, dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and crystallization was performed from diethyl ether-hexane to obtain 140 mg of 2-hydroxymethyl-3-phenylpropionic acid as a white solid. The optical purity is 98.0% e. e. Met. Melting point: 67.5-68.5 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ ppm:
2.75 to 2.93 (m, 2H), 2.97 to 3.13
(M, 1H), 3.60 to 3.85 (m, 2H), 5.
14 (br s, 2H), 7.06 to 7.34 (m, 5
H) [α] _D ²⁰ : + 13.9 ° (c = 0.97, CHCl ₃ ).

[0033]

According to the method of the present invention, optically active 2-hydroxymethyl-3-arylpropionic acid and its enantiomer ester can be produced in one step with good optical purity.

Claims (8)

[Claims] 1. A compound represented by the general formula (1): (In the formula, Ar is an aryl group which may have a substituent,
R represents an alkyl group, an aralkyl group or an aryl group, and * represents an asymmetric carbon atom. ), A 2-hydroxymethyl-3-arylpropionic acid ester represented by the general formula (2): (In the formula, Ar and * have the same meanings as described above.)
And a method for producing an optically active 2-hydroxymethyl-3-arylpropionic acid and its enantiomer ester. 2. The method according to claim 1, wherein Ar is a phenyl group which may have a substituent. 3. The method according to claim 1, wherein Ar is a phenyl group. 4. The production method according to claim 1, wherein R is an alkyl group having 1 to 8 carbon atoms. 5. The production method according to claim 1, wherein R is a methyl group. 6. The production method according to claim 1, wherein the enzyme is an enzyme of microbial origin or an enzyme of animal origin. 7. The enzyme is Achromobacter.
obacter) genus Aspergillus (Aspergi
llus) genus Bacillus (Bacillus) genus Candida (Candida) genus, Chromobacterium (Chromobacterium) genus, Humicola (Humicola) genus, Mucor (Mucor) genus,
Penicillium (Penicillium) genus Pseudomonas (Pseudomonas) genus and Rhizopus (R
The production method according to claim 1, wherein the enzyme is derived from a microorganism of the genus Hizopus ), or is derived from pig pancreas or bovine pancreas. 8. The enzyme is a chromobacterium ( Chro
genus mobacterium , Pseudomonas ( Pse
The method according to claims 1 to 5, wherein the enzyme originates from a microorganism of the genus Udomonas ) or porcine pancreas.