JPS58190395A - Preparation of optically active cis-imidazolidinedicarboxylic acid derivative - Google Patents

Abstract

PURPOSE:To prepare an optical active half ester useful as an intermediate for synthesizing biotin and other drugs by the action of a bacterium belonging to the genus Chromobacterium. CONSTITUTION:A bacterium such as Chromobacterium chocolatum (IFO3758) belonging to the genus Chromobacterium, capable of producing esterase, or esterase prepared by the bacterium capable of producing esterase is brought into contact with a cis-imidazolidinedicarboxylic diester shown by the formula II (R is lower alkyl; Bzl is benzyl), to give an optical active cis-imidazolidinedicarboxylic half ester shown by the formula I (R and Bzl are as shown above; the coordination of 4-position of imidazolidine ring is 4S, and that of 5-position is 5R).

Description

Detailed Description of the Invention The present invention is based on the general formula (I) (wherein R represents a lower alkyl group. Bzl represents a benzyl group. The coordination at the 4th and 5th positions is 4S and 5R coordination. The present invention relates to a method for producing the optically active cis-imidazolidinedicarboxylic acid derivative shown in 6).

That is, the present invention relates to cis-imidazolidine dicarboxylic acid diesters (hereinafter abbreviated as diesters) represented by the general formula (3) (wherein R represents a lower alkyl group and Bzl represents a benzyl group). t
') The optically active cis-imidazolidine dicarboxylic acid derivative represented by the above general formula (hereinafter abbreviated as Hafestel).

In general formulas (1) and (to), lower alkyl group means 01-6 alkyl group such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, n-hexyl group, etc. do.

Optically active halfesters (D
is an extremely important compound as a synthetic intermediate for biotin and other pharmaceuticals.

The present inventors have intensively investigated a method for producing optically active half-nystyl compounds (1) from diesters (6) using microorganisms. As a result, Chromobacterium (Chromobacterium
It has been found that optically active halfesters (I) can be obtained when an esterase derived from a Le: I have completed the method.

In the method of the present invention, when the hemihydrolysis is carried out by contacting with Chromobacterium, the reaction proceeds in good yield under mild conditions, and the objective compound of the present invention, Hafesters (I), can be obtained. The raw water decomposition reaction proceeds stereoselectively or specifically.

That is, when semi-hydrolyzing diester (6), which is a meso form, it is possible that two types of optical antipodes are produced, but if the method of the present invention is used, one of these two types can be produced. This results in so-called asymmetric hydrolysis, in which only the palmar form is produced in excess, and in some cases only the stereogenic form of one rice species is produced unilaterally.

The halfesters (I) selectively obtained by the method of the present invention have a 48.5R coordination and can be converted into optically active d-biotin. For example, one example is shown in reaction formula (a).

Reaction formula (4) % formula %() () (In the formula, R and Bzl are as described above, R1 represents a lower alkyl group, and X represents a halogen atom of C/, Br, or I.) That is, the optically active lactone (I) can be obtained by selectively reducing the lower alkoxycarbonyl group of the optically active halfester compound (I) and then ring-closing it. As a method for selectively reducing the lower alkoxycarbonyl group of the optically active hafesters (I), a reducing agent capable of selectively reducing the lower alkoxycarbonyl group, such as sodium borohydride, lithium borohydride, Diisobutylaluminum hydride (DI
BAL-El), sodium diethylaluminum hydride (OMI-1-1), and the like. The reaction solvent is not particularly limited as long as it does not hinder the progress of the reaction, but preferred solvents include ether solvents such as diethyl ether, tetrahydrofuran, dioxane, ethylene glycol, and dimethyl ether, and hydrocarbon reducing agents such as toluene and hexane. In this case, an alcoholic solvent such as ethanol or isopropanol, or a mixed solvent of these and water may be used. The reaction temperature may be room temperature, but it may be cooled below room temperature or heated to the boiling point of the solvent. When the reducing agent is an aluminum-based reducing agent, it is preferable to cool it to -70 to -20°C.

After the reduction reaction is completed, the reaction solution is made neutral or acidic, and then
Extract with organic solvents such as chloroform and ethyl acetate,
At this time, the ring is closed and an optically active lactone can be obtained.

In order to obtain optically active lactones in good yield, it is preferable to make the reaction solution acidic after the reduction reaction.

The lactone (I) is a known compound, and is derived into the corresponding thiolactone (IV). Next, the alcohol M obtained by introducing a side chain by Grignard reaction is dehydrated and hydrogenated to obtain a compound (good). This compound (V
I) was converted into a thiophanium salt (■) with hydrogen halide, reacted with diethyl malonate to form a compound (■), and then hydrolyzed and decarboxylated to remove the Nl and N3-dibenzyl groups to form optically active d. -Biotin (TX) can be obtained.

Therefore, the optically active halfesters α) obtained by the present invention can be led to optically active d-biotin (IX) without undergoing any special optical resolution step and without undergoing racemization during the process. This makes possible an extremely advantageous industrial production method for d-biotin (ff).

To carry out the method of the present invention, Chromobacterium (Chromobacterium
(obactarium) gll, and the diesters (2
), continue culturing, and perform semi-hydrolysis. In this case, there is no particular restriction on the culture temperature as long as the growth of bacteria is possible, but ii! It is preferable to carry out the culturing at a temperature of s'125 to 87°C, the number of days of culturing is suitably between half a day and 7 hours, and the substrate concentration is suitably about 0.1 to 1096.

A method of collecting microorganism culture fluid by centrifugation or other operations and performing semi-hydrolysis using the microbial cells, or a method of releasing enzymes by subjecting microorganism cells to ultrasonic treatment, lysozyme treatment, etc., followed by centrifugation under cooling, It can also be carried out by obtaining an esterase by ammonium sulfate fractionation, precipitation dialysis, etc. and performing semi-hydrolysis using this. In these cases, general formula (2)
The diesters represented by are suspended or dissolved in an aqueous solution or a buffer solution preferably adjusted to pH 5 to 8, and microbial cells or esterase are added to the diesters at a weight ratio of preferably o, oot to 0.1. The reaction proceeds preferably by stirring at 10 to 40°C, and is usually completed in 1 hour to several days.

As mentioned above, the aqueous solution includes sulfuric acid, hydrochloric acid, phosphorous mineral acid, acetic acid,
Organic acids such as citric acid, inorganic bases such as sodium hydroxide, potassium hydroxide, and sodium carbonate, triethylamine,
It refers to an aqueous solution containing an organic base such as pyridine and a salt such as sodium acetate or sodium chloride.

Slow percent solution means common buffer solutions of potassium dihydrogen phosphate-sodium hydroxide, potassium dihydrogen phosphate-sodium dihydrogen phosphate, potassium hydrogen phthalate-hydrochloric acid, glycine-sodium chloride-sodium hydroxide. However, there are no particular restrictions other than those that can prevent reactions.

Furthermore, when carrying out the present invention, methanol, ethanol, n-propertool, isopropertool, n-
Alcohol solvents such as butanol and t-butanol,
Ether solvents such as ether, tetrahydrofuran and dioxychloride; aromatic hydrocarbon solvents such as benzene and toluene; ketone solvents such as acetone, methyl ethyl ketone and diethyl ketone; amine solvents such as triethylamine and pyridine; and dimethylformamide. , dimethyl sulfoxide, a polar solvent, and sorbitan monopalmitate,
Surfactants such as sorbitan monolaurate and the like can also be added.

1.8-Dibenzyl 2-oxoimidazolidine-4t5
- Diesters obtained by dehydrating dicarboxylic acids and methanol, ethanol, n-propanol, isopropanol, n-phthanol, t-butanol, n-pentanol, or n-hexanol in the presence of sulfuric acid. It can be synthesized as (to).

The present invention will be explained in more detail below using practical examples, but it goes without saying that the present invention is not limited thereto.

Reference Example 1 Production method of cis-1,8-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylic acid diethyl ester Cis-1,8,-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylic acid 985. 41, Benzene 500
-199.596 ethanol 4fi clear/concentrated EN acid 5.
After refluxing the reaction solution for 10 hours, the reaction solution was cooled, and the reaction solution was washed with water, 5% NaOH water, and water in this order.

Thereafter, it was concentrated under reduced pressure, and the resulting residue was recrystallized from 95% ethanol.

m-P 74-76°C NoizX 1.8') Hensyl-2-oxoimidazolidine-4,5-dicarboxylic acid diethyl ester was obtained.

Example 1 Glucose 1.0? , yeast extract (Kyokuto Pharmaceutical Industries)
0.5F, Peptone (Mikuni Chemical Industry) 1.0P,
Dipotassium hydrogen phosphate 0. FI F, distilled water 100-
The pH of the solution was adjusted to 7.0 with dilute hydrochloric acid. After steam sterilizing this liquid culture solution at 120'C for 20 minutes,
Chromobacterium Chooolatu
m (IFθ3758) was inoculated and cultured with shaking at 26°C for 48 hours.

To this, cis-1,8-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylic acid dimethyl ester 400
After adding ~, the reaction was further carried out with shaking at 26°C for 72 hours.

After adding 100% of ethyl acetate to this reaction solution and making it acidic with diluted hydrochloric acid zeta, it was poured onto celite.

The organic layer was washed with water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure.

5*1ill 20ml of water was added to the residue, and after dissolution, it was washed with 20ml of ether and separated.

The water droplets were adjusted to pH 2 with dilute sulfuric acid, and the precipitated white crystals were filtered off, washed with water, and then dried.

(48,5K) -1,8-') Henry's -5-methoxycarbonyl-2-oxoimidazolidine-4-carboxylic acid 84011F was obtained.

Melting point 149-151°C [a] ass -28,0° (C=1.DMF)
Met.

Reference example 2 Lithium hydride 4819 and tetrahydrofuran 8
-(4B,5R)-1 obtained in Example 1,
A solution consisting of 240 wq of 8-dibenzyl-5-methoxycarbonyl-2-oxoimidazolidine 4-carboxylic acid and 4 wd of tetrahydrofuran was added dropwise at room temperature.

Next, this reaction solution was refluxed for 2 hours, and then tetrahydrofuran was distilled off. While cooling, 4-4 methanol was gradually added to the residue, and then 0.5 i of concentrated hydrochloric acid was added. This mixture was refluxed for 1 hour and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain (4aS, 5aR)-t, 8-dibenzylhexahydro-IH-furo(8,4-d]imidazole-2,4-dione.

Melting point 117-118°C [a]n +62.5° (C,=t, (l[[/
3) Example 2 Glucose8. OF, yeast extract (Kyokuto Pharmaceutical Industries) 1
．． 5 F, Peptone (Mikuni Chemical Industry) 8.0? A solution consisting of 1.5 F dipotassium hydrogen phosphate and 800 mg of distilled water was adjusted to pH 47.0 with dilute hydrochloric acid. After sterilizing this culture solution with steam for 20 minutes in a hot iron at 120°C,
mobacterium Chocolatum (I
FO8758) and cultured with shaking at 26°C for 72 hours.

The culture solution was centrifuged at 5,000 rpm for 25 minutes using a cooler to collect bacteria. The bacterial cells were washed 8 times with 100 m of 0.8611 saline (by centrifugation) and collected. 0.1 for bacterial cells
M-phosphate buffer solution (PE[7,0) 50m, cis-1,8-dibenzyl-2-oxoimidazolidine-4
, 5-dicarboxylic acid dimethyl ester (1.50 P) was added thereto, and the mixture was stirred at room temperature for 70 hours while adjusting the pH to around 7 with a 1N water furnace sodium solution.

This reaction solution was centrifuged and separated into bacterial cells and supernatant (M cells were collected). The supernatant liquid was adjusted to pH 2 with dilute sulfuric acid, and the precipitated white crystals were filtered, washed with water, and then dried.

(4s, 5fL)-1+ 8-dibenzyl-5-methoxycarbonyl-2-oxoimidazolidine-♦-carboxylic acid L40F was obtained.

Melting point 145-147°C [α]ars 25.0° (C=1.DMF
) Reference Example 8 A mixture of sodium borohydride 820~ and isopropyl alcohol 85~ obtained in Example 2 (4B, 5R
)-1,8-dibenzyl-5-methoxycarbonyl-2
A solution consisting of 1.20 P of -oxoimidazolidine-4-carboxylic acid and 15 m of tetrahydrofuran was added dropwise at room temperature. Next, this reaction solution was stirred at 60 to 70°C for 7 hours. The reaction solution was cooled, 2.51% of W# hydrochloric acid was added, and the reaction solution was concentrated under reduced pressure. 250 μl of water was added to the residue, and the mixture was extracted twice with 100 μl of chloroform. The chloroform layer was washed with water, dried over anhydrous magnesium silicate, and concentrated under reduced pressure to obtain a residue of 1.08F.

The residue was subjected to silica gel column chromatography fPIa (4 m8
．． 6aK) -1,8-dibenzylhexahydro-I
H-furo[B,4-d]imidazole-2,4-dione was obtained.

Melting point 115-118°C [“)D+59.5° (C==1, C:HCl5
) Example 8 '46 Two bacterial cells collected in Example 2 were treated with 0.1 M-IJ acid buffer resistance (PH'i', o) 50 ynt, cis-1
,8-dipenzyl-2-oxoimidazolidine-4,5
-dicarboxylic acid dimethyl ester 1.50? was added and treated in the same manner as in Example 2 to obtain (48,5R)-1,3-dibenzyl-5-methoxycarbonyl-2-oxoimidazolidine-4-carboxylic acid 184F.

Melting point: 146-148℃

Claims (1)

[Claims] General formula (in the formula, R represents a lower alkyl group. Bzl represents a benzyl group) - To the cis-imidazolidine carboxylic acid diesters represented by D, to the genus Chromobacterium The general formula (wherein R represents a lower alkyl group. Bzl represents a benzyl group. The method for producing an optically active cis-imidazolidinedicarboxylic acid derivative represented by