JPH0475587A - Production of rhododendrol - Google Patents

Abstract

PURPOSE:To produce (+)-rhododendrol having high optical purity on an industrial scale without being influenced by weather, seasons, etc., by treating 4-(p- hydroxyphenyl)-2-butanone with yeast. CONSTITUTION:Yeast is cultured in a medium containing glucose, corn steep liquor and peptone, etc., for about 24-48 hours and mixed with about 0.5g/l-50g/l4-(p-hydroxyphenyl)-2-butanone of a starting substance to start reaction. The reaction time varies with an amount of 4-(p-hydroxyphenyl)-2-butanone added and generally requires 1-7 days. Rhododendrol thus formed is recovered by separating the cell from the medium and extracting the medium and the cell with an organic solvent. The extracted solutions are joined, concentrated under reduced pressure, rhododendrol is purified by silica gel column chromatography and recrystallized to give rhododendrol.

Description

Detailed Description of the Invention The present invention utilizes yeast to produce 4-(p-hydroxyphenyl)-2-butanone (4-(#-hydroxyphenylene)
The present invention relates to a method for producing rhododendrol from nyl)-2-butanone). Rhododendrol is
It is a substance that is expected to be used as a medicine, especially as a recovery drug for liver diseases.

Subordinate Branch (+) - Rhodopentrol is a plant of the genus Rhododendoron maximu and Rhododendoron maximu.
It is a substance that is known to exist in M. and was isolated and identified as one of the components that exhibits a protective effect against experimental liver damage from Megusurinoki [Shinoda et al., Journal of Pharmacy, 40 (2) 1
77-181 (1986)).

Megusurinoki is a deciduous tree of the maple family that is native to Japan.

This plant is widely distributed throughout Japan except Hokkaido and Okinawa, but because it grows wild in the mountains at an altitude of around 700 m and grows slowly, it is limited in terms of resources.

Therefore, it is practically impossible to collect such plants in large quantities, and therefore it is also impossible to obtain large amounts of (+)-rhodopuntrol from these plants.

For the above-mentioned reasons, the present inventors first wanted to develop a method for producing a large amount of Megusurinoki by culturing and propagating callus derived from the living tissue of Megusurinoki.
In order to obtain a large amount of rhododendrol by conventional callus culture, it is necessary to culture a large amount of callus and then separate and purify it. For this reason,
There is a problem in that it is difficult to efficiently obtain large amounts of the above substances by conventional callus culture. On the other hand, the present inventors would like to develop a method for efficiently producing rhododendrol from its precursor by utilizing callus derived from the living tissues of the Japanese Patent Application No. 2-45287.
Publication No.). However, with this method, there is a limit to the amount of precursor material that Megusurinoki callus can convert at one time, and in order to obtain a large amount of rhododendrol, the conversion must be repeated many times, resulting in a large amount of time. There is a problem.

Rhododendoron @axi
In Japan, it is impossible to stably obtain large amounts of (+)-rhododendrol due to regional, seasonal and weather influences.

＜”´ I will do it.

Taking these points into consideration, the present invention aims to produce rhododendrol, which is expected to be used as a pharmaceutical, stably and in large quantities without being affected by regional, seasonal, or weather conditions. That is, an object of the present invention is to provide a method for producing rhododendrol more efficiently than 4-(p-hydroxyphenyl)-2-butanone.

The feature of the present invention is to make use of the ketone reducing ability of yeast to
The purpose of the present invention is to convert (p-hydroxyphenyl)-2-butanone into rhododendrol.

Rhododendrol as used herein is 4-(p-hydroxyphenyl)-2-butanol, and is represented by the following structural formula. This compound can be obtained as the (±) form by reacting 4-(p-hydroxyphenyl)-2-butanone with an appropriate reducing agent, but it is difficult to separate the (+) and (-) H forms. It is. When using yeast,
Prelog heuristics [V, Prelog, Pure
Appl, Cheap.

9, 119 (1964)], the possibility of selective production of the (decade) isomer is higher.

In the present invention, 4-(p-hydroxyphenyl)-2-phthanone is added to a yeast culture solution to convert it into rhododendrol.

4-(p-hydroxyphenyl)-2-butanone, which is the starting material of the present invention, is a known substance and can be easily obtained because it is industrially produced.

As a yeast, Saccharomyces cerevisiae (Sac
charomyces cerevisiae), Saccharo@yces
baiLii), etc.

For culture, yeast is inoculated into a medium containing appropriate carbon sources, nitrogen sources, vitamins, etc.
-(p-hydroxyphenyl)-2-butanone is added. More specifically, yeast was grown for 24 hours in a medium containing glucose, corn steep liquor, peptone, etc.
After ~48 hours of incubation, the starting material 4-(p-hydroxyphenyl)-2-butanone was added at 0.5 g/12-50 g.
/ 1. , preferably 1.0 g/f to 10 g/f, to start the reaction. 4-(p-hydroxyphenyl)
Since -2-butanone is sparingly soluble in water, it is desirable to dissolve it in a small amount of an organic solvent that does not affect the growth of yeast, such as ethanol, acetone, etc. before adding it. The reaction time varies depending on the amount of 4-(p-hydroxyphenyl)-2-butanone added, but generally takes 1 to 7 days. Rhododendrol produced by such conversion is recovered by separating the culture medium and cells and then extracting each with a solvent. The solvents used include acetone, methanol, ethanol, ether, Examples include ethyl acetate. After the extracts are combined and concentrated under reduced pressure, rhododendrol can be obtained by performing silica gel column chromatography or the like to purify rhododendrol and recrystallizing it.

Note that yeast can be used repeatedly by immobilizing it on a suitable support material.

Hereinafter, the present invention will be specifically explained with reference to Examples.

Example (1) Dried baker's yeast Saccharomyces cerevisiae was added to an Erlenmeyer flask on a Rodin roll.
e) (made by Oriental Yeast ■) 10g, glucose 5g
Then, 50 d of tap water was added, and the mixture was shaken reciprocally for 10 minutes at room temperature (amplitude: 4C, 1,130 reciprocations/min). After that, 0.2
4Ap-hydroxyphenyl)-2-butanone (manufactured by Tokyo Kasei Kogyo ■) dissolved in ethanol was added to
3 mmol) and reacted for 27 hours. In addition,
During the reaction period, yeast (2 g) was added at 16.5 and 20 hours, and glucose (2 g) was added at 2.5, 16.5 hours. and 2
Each was added at 0 hours.

(2) After completion of the reaction with the rhododendron, the cells were extracted with acetone (100adx2) and the medium was extracted with diethyl ether (looIII1X3). The extracts were combined and concentrated under reduced pressure, and then subjected to silica gel column chromatography (using tohexane and diethyl ether) to isolate rhododendrol. The yield was 63%.

(3) The produced rhododendrol was subjected to high performance liquid chromatography (HPLC) analysis using a column for optical isomer separation to measure its optical purity. The measurement conditions are as follows.

Column: CHIRALCEL 00 (φ4.6
X250) (manufactured by Daicel Chemical Industries, Ltd.) Mobile phase: 3% isopropylamine [n-hexane-2-
Proper tool (95:5)) Flow rate = 1 d/in Temperature: 20°C Detection: LIV 277 ns+ From these results, it was found that optical purity (+)-Rhodometol was obtained with an optical purity of 95% or more.

FIG. 1 shows an example of HPLC analysis of synthetic rhododendrol and rhododendrol produced by this method.

According to the method of the present invention, rhododendrol is obtained from 4-(p-hydroxyphenyl)-2-butanone using yeast, so it can be produced on an industrial scale without being affected by weather, seasons, etc. can produce (+)-rhodopuntrol with high optical purity.

Claims (3)

[Claims] (1) A method for producing rhododendrol, which comprises converting 4-(p-hydroxyphenyl)-2-butanone into rhododendrol by the action of yeast. (2) Claim (1) characterized in that the conversion to rhododendrol is carried out by adding 4-(p-hydroxyphenyl)-2-butanone to a yeast culture solution and causing a reaction.
The method for producing rhododendrol described in . (3) Yeast is Saccharomyces cerevisiae (Saccharomyces cerevisiae)
The method for producing rhododendrol according to claim (1) or (2), wherein the rhododendrol is A. aromyces cerevisiae.