JP3203865B2 - Method for producing acetylene alcohol compound - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing an acetylene alcohol compound for obtaining a compound useful as an intermediate for medicines, agricultural chemicals and liquid crystals.

[0002]

2. Description of the Related Art A method for synthesizing an optically active acetylene alcohol compound according to the present invention is disclosed in Japanese Unexamined Patent Publication No. Hei 3-20199.
No. 6 describes the following method.

[0003]

In the above-mentioned conventional method, a hydrolysis enzyme is reacted with an alcohol-organic solvent to carry out alcoholysis. However, although a relatively high optical purity is achieved in the obtained ester, the hydrolyzed alcohol is used. There is no description of optical purity or the like. According to the method, the ester is further chemically hydrolyzed to obtain an alcohol having a relatively high optical purity. In general, in order for the ester remaining as a hydrolysis residue to have a high optically active purity, it is necessary to increase the hydrolysis rate. As a result, the optical purity of the alcohol becomes insufficient.

[0005]

SUMMARY OF THE INVENTION
The present inventors have conducted intensive studies on a method for obtaining one enantiomer of an acetylene alcohol ester compound with good optical purity and high yield.As a result, the asymmetric hydrolysis of a racemic acetylene alcohol ester compound using an esterase was carried out. The present inventors have found that an optically active acetylene alcohol compound having a high purity can be obtained, and have reached the present invention.

[0006]

That is, the present invention provides a compound represented by the general formula [1]: (In the formula, R 1 is an optionally halogenated carbon number 1
Represents an alkyl group of 5 to 5, and n represents an integer of 2 to 6. )
Is subjected to asymmetric hydrolysis using an esterase having the ability to preferentially hydrolyze the R-form of the optically active form of the acetylene alcohol ester compound represented by the general formula [2]

[0007] (In the formula, n represents the same meaning as described above, and * represents an asymmetric carbon atom.) And an optically active acetylene alcohol compound (R-form) represented by the general formula [3]:

[0008] (In the formula, R1, n and * represent the same meaning as described above.)
And a method for obtaining an optically active acetylene alcohol ester compound (S form) represented by

Hereinafter, the present invention will be described in detail. In the present invention, examples of the acetylene alcohol ester compound represented by the general formula [1] as a raw material include, for example, 5-hexyn-2-ol, 6-heptin-2-ol, 7-octin-2-ol, 8-octin-2-ol. The following esters such as nonin-2-ol and 9-decin-2-ol can be exemplified. Esters such as acetic acid, propionic acid, butanoic acid and pentanoic acid which may be substituted by a fluorine atom, a chlorine atom, a bromine atom and the like.

The esterase having the ability to selectively hydrolyze the R- isomer of the optical isomer of the acetylene alcohol ester compound in this reaction means an esterase in a broad sense including lipase. Among them, lipase is preferred,
Still more specifically, lipases derived from the genus Arthrobacter and Pseudomonas are exemplified. In this reaction, an enzyme obtained from an animal, a plant, or a microorganism is used as the esterase, and the use form thereof is a purified enzyme, a crude enzyme, an enzyme-containing substance, a microorganism culture solution, a culture, a cell, a culture filtrate, Alternatively, they can be used in various forms as required, such as those obtained by treating them, and an enzyme and a microorganism can be used in combination. Alternatively, it can be used as an enzyme immobilized on a resin or the like, or immobilized cells.

[0011] Microorganisms producing esterase include:
The microorganism is not particularly limited as long as it is a microorganism that produces an esterase having the ability to selectively hydrolyze the R-form of the optical isomer of the acetylene alcohol compound. Specific examples of such microorganisms include, for example, genus Enterobacter, Arthrobacter, Brevibacterium,
Pseudomonas, Alcaligenus, Micrococcus, Bacillus, Lactobacillus, Trichoderma,
Candida spp. , Hansenula,
Microorganisms belonging to the genus Achromobacter are exemplified. The cultivation of the microorganism is generally performed according to a conventional method. For example, a culture solution can be obtained by performing liquid culture.

For example, a sterilized liquid medium [malt extract / yeast extract medium for molds and yeasts (5 g of peptone, 10 g of glucose, 3 g of malt extract, 3 g of yeast extract, and 3 g of yeast extract in 1 L of water) is adjusted to pH 6.5. ), For bacteria, inoculated with microorganisms in a sweetened bouillon medium (5 g of peptone, 10 g of glucose, 3 g of meat extract, and 3 g of NaCl dissolved in 1 L of water to have a pH of 7.2), and usually at 20 to 40 ° C for 1 to 3 It is performed by culturing for days, and solid culturing may be performed if necessary. Some of these microbial esterases are commercially available and can be easily obtained. Specific examples of commercially available esterases include, for example, lipases of the genus Pseudomonas [Lipase Amano PS and Lipase Amano P (both manufactured by Amano Pharmaceuticals)] and lipases of the genus Aspergillus [Lipase Amano AP, Lipase Amano A12
(Both manufactured by Amano Pharmaceutical Co., Ltd.)], a lipase of the genus Mucor [Lipase Amano M-AP (manufactured by Amano Pharmaceutical)], a lipase of Candida cylindrasse [lipase MY (manufactured by Meito Sangyo)], a lipase of the genus Alcaligenes [lipase PL]
Lipase of Acromobacter sp. [Lipase AL (manufactured by Meito Sangyo)], lipase of Arthrobacter sp. [Nippon Chemical], lipase of chromobacterium sp. (Toyo Brewery), Lipase of Rhizopus delemar [Taripase (manufactured by Tanabe Seiyaku)];

[0013] Animal / plant esterases can also be used. Specific examples of these esterases include stearpsin, pancreatin, pig liver esterase,
Wheat Germ esterase and the like can be mentioned. The amount of the enzyme is usually 0.001 to equivalent times, preferably 0.003 to 0.5 times the acetylene alcohol ester compound [1]. Of course, more than this amount may be used. The asymmetric hydrolysis reaction is generally carried out by vigorously stirring a mixture of an acetylene alcohol compound and the above enzyme or microorganism in a buffer solution. Examples of the buffer include commonly used buffers of inorganic acid salts such as sodium phosphate and potassium phosphate, and buffers of organic acid salts such as sodium acetate and sodium citrate. PH 8-11 is preferable for the culture solution and the alkaline esterase, and pH 5-8 is preferable for the culture solution of the non-alkaliphilic microorganism and the esterase having no alkali resistance. Concentration is usually 0.05
-3M, preferably in the range of 0.05-0.5M.

The reaction temperature is usually 10-60 ° C., and the reaction time is generally 1-30 hours, but is not limited thereto. When a lipase belonging to the genus Pseudomonas or the genus Arthrobacter is used as the lipase in this asymmetric hydrolysis reaction, an optically active acetylene alcohol compound with relatively high optical purity can be obtained.
In addition, at the time of this asymmetric hydrolysis reaction, in addition to the buffer solution, an organic solvent inert to the reaction such as toluene, chloroform, methyl isobutyl ketone, and dichloromethane can be used. Can be done. The course of the reaction can be monitored by monitoring the optical purity of the optically active acetylene alcohol compound [2] in the reaction system, for example, by liquid chromatography or the like provided with a filler for separating the optically active compound, and thereby the reaction end point You can also decide. After completion of the reaction, if necessary, hexane, heptane, benzene, acetone, toluene, dichloromethane, chloroform, chlorobenzene, dichloroethane, ethyl acetate, an aliphatic or aromatic hydrocarbon such as ethyl ether, ether, ketone, ester, etc. After adding a solvent such as a halogenated hydrocarbon, for example, filtering off the enzyme, the filtrate is concentrated to obtain an optically active acetylene alcohol compound represented by the general formula [2] and an optically active acetylene represented by the general formula [3] An alcohol ester compound can be obtained. Also,
Further, for example, by performing a usual chromatography treatment, it can be separated into an optically active acetylene alcohol compound represented by the general formula [2] and an optically active acetylene alcohol ester compound represented by the general formula [3].

The optically active acetylene alcohol ester compound [3] thus obtained can be converted into an enantiomer optically active acetylene alcohol compound by further hydrolysis. In the acetylene alcohol compound represented by the general formula [1], when R ¹ is an alkyl group which may be halogenated having 2 to 5 carbon atoms, acetylene which is optically active with relatively high optical purity is used. An alcohol compound [2] can be obtained.

[0016]

According to the method of the present invention, the optically active acetylene alcohol compound represented by the general formula [2] and the optically active acetylene alcohol ester compound represented by the general formula [3] can be obtained in a relatively short time and with a high optical activity. It is industrially advantageous because it can be obtained in high yield and high yield.

[0017]

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

Example 1 6-Heptin-2-ol
18.2 g of butanoic acid ester, Arthrobacter lipase
(Nippon Chemical) 0.36g, 0.2M phosphate buffer
-(PH 6.0) Vigorously mix 100 ml of the mixture at 30 ° C for 14 hours
Stir. The resulting mixture is ethyl ether 100 m
The organic layer is washed with water and concentrated. Obtained residue
Checking is performed by silica gel column chromatography (eluent:
Chloromethane-hexane), and R (-)-6-
4.0 g of Putin-2-ol (optical purity 95.4%)
[Α]_D ²⁰: -14.5 ° (c = 5, benzene) and S
Butanic acid ester of (+)-6-heptin-2-ol
Get. The optical purity of the compound
Liquid treated with optically active column after treatment with socyanate
It was measured by body columnography.

Example 2 6-Heptin-2-ol
5.7 g of 4-chlorobutanoic acid ester, Lipase Amano PS
(Manufactured by Amano Pharmaceutical) 0.2 g and 0.3 M phosphate buffer
(PH 6.5) Vigorously stir 60 ml of the mixture at 30 ° C for 8 hours
I do. The resulting mixture is ethylene dichloride 30 ml
The organic layer is concentrated after washing with water. Obtained residue
Is a silica gel column chromatography (eluent: dic
Hexane), and R (-)-6-hep
1.06 g of tin-2-ol (optical purity 96.8%)
[Α]_D ²⁰: -15 ° (c = 5, benzene) and S
4-chlorobutane of (+)-6-heptin-2-ol
An acid ester is obtained.

Example 3 3.85 g of acetate of 6-heptin-2-ol, Lipase Amano PS (manufactured by Amano Pharmaceutical Co., Ltd.)
0.1 g and 0.2 M phosphate buffer (pH 6.0) 50
Stir vigorously the ml mixture at 30 ° C. for 10 hours. The obtained mixture is extracted with 100 ml of ethyl ether, and the organic layer is washed with water and concentrated. The obtained residue was separated by silica gel column chromatography (eluent: dichloromethane-hexane) to give R (-)-6-heptin-2-ol.
0.56g (optical purity 72%) Optical rotation [α] _D ²⁰ : -11 °
(C = 5, benzene) and S (+)-6-heptin-2
To obtain the acetate of the ol.

Example 4 3-Nonon-2-ol-3
Chloropropionate 5.7 g, lipase amano
A mixture of 0.2 g of PS (manufactured by Amano Pharmaceutical) and 60 ml of 0.2 M phosphate buffer (pH 7) is vigorously stirred at 30 ° C. for 10 hours. The obtained mixture is extracted with 60 ml of ethyl ether, and the organic layer is washed with water and concentrated. The obtained residue was separated by silica gel column chromatography (eluent: dichloromethane-hexane), and R (-)-8-nonine-2-
All 1.22 g (optical purity 94.5%) Optical rotation [α] _D ²⁰ :-
11.7 ° (neat) and the 3-chloropropionate of S (+)-8-nonin-2-ol are obtained.

────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-3-247299 (JP, A) JP-A-3-259094 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) C12P 41/00 BIOSIS (DIALOG)

Claims (3)

(57) [Claims] 1. The general formula [1] (In the formula, R 1 is an optionally halogenated carbon number 1
Represents an alkyl group of 5 to 5, and n represents an integer of 2 to 6. )
Asymmetric hydrolysis using an esterase having the ability to preferentially hydrolyze the R-form of an optically active acetylene alcohol ester compound represented by the general formula [2]: (In the formula, n represents the same meaning as described above, and * represents an asymmetric carbon atom.) And an optically active acetylene alcohol compound (R-form) represented by the general formula [3]: (Wherein, R1, n and * represent the same meanings as described above.) A process for producing an optically active acetylene alcohol ester compound (S-form) represented by the formula: 2. The method according to claim 1, wherein the esterase is an enzyme belonging to the genus Arthrobacter or Pseudomonas. 3. In the above general formulas [1] and [3],
R ¹ is The process of Claim 1 which is an alkyl group which may be halogenated 2 to 5 carbon atoms.