JPH0482135B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is based on the formula a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
This invention relates to a 3'',6'-dihydroxy-ML-236B derivative and a method for producing the same. Conventionally, ML-236B derivatives in which the hydroxy group at the 3'' position in the above general formula () is substituted with a hydrogen atom have been disclosed in, for example, JP-A-57- No. 2240, No. 57-50894,
No. 57-67575, No. 57-155995, No. 58-10572
No. 58-89191, etc., and
MB-530B in which the 6′-hydroxyl group is substituted with a methyl group
Derivatives are described in U.S. Pat. No. 4,376,863;
All are known to exhibit cholesterol synthesis inhibitory effects. The present inventors have completed the present invention by discovering that a carboxylic acid having the general formula (), its pharmacologically acceptable salt, its ester, or its ring-closed lactone form all exhibit a cholesterol synthesis inhibitory effect. Examples of the pharmacologically acceptable salts of the compound having the general formula () of the present invention include metal salts, amino acid salts, and amine salts. Examples of metal salts include alkali metal salts such as sodium and potassium, alkaline earth metal salts such as calcium and magnesium, aluminum salts, iron salts, zinc salts, copper salts, nickel salts, and cobalt salts. Among them, alkali metal salts, alkaline earth metal salts and aluminum salts are preferred, and sodium salts, potassium salts, calcium salts and aluminum are most preferred. Examples of amino acid salts include arginine, lysine, histidine, α,
Basic amino acids such as γ-diaminobutyric acid and ornithine are preferred. Examples of amino salts include t-
Preferred are octylamine, dibenzylamine, dicyclohexylamine, morpholine, D-phenylglycine alkyl ester, D-glucosamine, and the like. Examples of the ester of the compound having the general formula () include alkyl esters such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and bentyl. Methyl is preferred. The ring-closed lactone of the compound having the general formula () above refers to a compound in which the formula () is represented by the following ring-closed structural formula. Examples of the carboxylic acid having the general formula (), its pharmacologically acceptable salt, its ester, or its ring-closed lactone derivative obtained by the present invention include the compounds described below. (1) 3″,6′-dihydroxy-ML-236B carboxylic acid (2) 3″,6′-dihydroxy-ML-236B carboxylic acid sodium salt (3) 3″,6′-dihydroxy-ML-236B carboxylic acid Potassium salt (4) 3″,6′-dihydroxy-ML-236B carboxylic acid calcium salt (5) 3″,6′-dihydroxy-ML-236B carboxylic acid aluminum salt (6) 3″,6′-dihydroxy-ML -236B carboxylic acid arginine salt (7) 3″,6′-dihydroxy-ML-236B carboxylic acid lysine salt (8) 3″,6′-dihydroxy-ML-236B carboxylic acid t-octylamine salt (9) 3″ ,6′-dihydroxy-ML-236B carboxylic acid D-phenylglycine ethyl ester salt (10) 3″,6′-dihydroxy-ML-236B carboxylic acid methyl ester (11) 3″,6′-dihydroxy-ML- 236B Lactone Form In the general formula () of the present invention, various geometric isomers exist depending on the arrangement of substituents. Also, various optical isomers exist due to the presence of an asymmetric carbon atom.The general formula ( ), in
All of these isomers and mixtures of these isomers are shown in a single formula. Therefore, in the present invention, the carboxylic acid having the general formula (), its pharmacologically acceptable salt, its ester, or its ring-closed lactone includes not only these isomers but also mixtures of these isomers. It also includes all. The object compound of the present invention has an effect of lowering blood lipids by inhibiting cholesterol synthesis, and can be used in medicine, for example, as a therapeutic agent for hyperlipidemia or an agent for preventing arteriosclerosis. These compounds can be administered orally or parenterally, for example in the form of capsules, tablets, injections and the like. The dosage varies depending on age, symptoms, body weight, etc., but it is usually about 0.2 to 200 mg per day per day.
mg is administered in 3 to 4 divided doses. However, larger amounts can be used if desired. The raw materials of the present invention, such as ML-236B lactone and ML-236B carboxylic acid, are known substances as described above, and are separated and purified from metabolites of Penicillium titrinum, a type of blue mold. Its chemical structure is as follows and As shown by It is known that it exhibits a strong serum cholesterol-lowering effect even at the individual level (Japanese Unexamined Patent Publication No. 155690/1983, Atherosclerosis, Vol. 32,
pp. 307-313, 1979). The object compound of the present invention has the formula A carboxylic acid having the following, a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
The ML-236B derivative is hydroxylated using a microorganism belonging to the genus Nocardia or the genus Streptomyces to convert it into a 3″,6′-dihydroxy-ML-236B derivative, and then 3 '',6'-dihydroxy-ML-236B derivative. The derivative thus obtained can be further converted, if desired, by subjecting it to a hydrolysis reaction, a salt formation reaction, an esterification reaction, or a lactonization reaction. Examples of microorganisms that can convert the raw material compound into a carboxylic acid having the general formula (), a pharmacologically acceptable salt thereof, an ester thereof, or a closed ring lactone thereof include the genus Nocardia and the genus Streptomyces. Among these microorganisms, especially Nocardia sp.
SANK62781 (Feikoken Bacteria No. 6181) Nocardia sp.
SANK62881 (Feikoken Bacteria No. 6182) Nocardia sp.
SANK62981 (FERM BP-1145) is preferred. Streptomyces carbophilus SANK62585 (FERM BP-1145). Among these strains, Nocardia sp.
SANK62781, SANK62881 and SANK62781
The mycological properties of SANK62981 have already been reported in JP-A-58-
Described in No. 89191. Streptomyces carbophilus
The mycological properties of SANK62585 strain are as follows. 1 Morphological characteristics The morphology is ISP (International Streptomyces progeny).
Streptomyces Project)] on specified medium, 28℃,
After culturing for 14 days, it was observed under a microscope.
The basal hyphae of the SANK62585 strain are branched and elongate well, and the aerial hyphae are simply branched. The shape of the spore chain of the SANK62585 strain is usually straight to curved, but it may also be spiral. The surface structure of the spore chain is smooth. In addition, special organs such as axle branches of aerial hyphae, sclerotia, fractures of basal hyphae, and sporangia were not observed. 2 Properties on various culture media Properties after culturing at 28°C for 14 days on various culture media are shown in Table 1. The color tone display represents the color chip number of the "Standard Color Chart" published by the Japan Color Research Institute. [Table] [Table] [Table] Sex pigment 3 Physiological properties The physiological properties of the SANK62585 strain are shown in Table 2. [Table] In addition, using Pridham-Gotlieb agar medium, carbon sources after 14 days of culture, namely D-glucose, L-arabinose, D-xylose, inositol, D-mannitol, D-fructose,
L-rhamnose, sucrose, raffinose,
The assimilation properties of cellobiose and trehalose were investigated.
Since the SANK62585 strain grows well even in a control medium without carbon source addition, it is difficult to describe its exact assimilation ability. However, significantly better growth was observed in the medium supplemented with D-glucose, D-xylose, inositol, raffinose, cellobiose, and trehalose compared to the non-additive control medium. 4 Regarding bacterial cell components The cell wall of SANK62585 strain was determined by the method of Becker et al. [B Becker et al., Applied Microbiology, Vol. 12,
421-423, 1964], L.
Since L-diaminopimelic acid and glycine were detected, it was confirmed that the cell wall was type I. In addition, sugar components in whole cells of the SANK62585 strain were analyzed using the MP Reciever method [MP
Lechevalier, Journal of Laboratory and Clinical Medicine
of Laboratory and Clinical Medicine), vol. 71,
934, 1968], no characteristic pattern was observed. From the above, it was determined that this bacterial strain belongs to the genus Streptomyces among actinomycetes. The SANK62585 strain was identified by ISP [The International Streptomyces Project].
Project)] Criteria; Bergey's Manual of Determinative
Bacteriology) 8th edition; The Actinomycetes, Volume 2 by SAWaksman, and recent literature on actinomycetes. As a result, this strain did not correspond to any known species of the genus Streptomyces, so it was determined that it was a new species of the same genus.
(FERM BP-1145). Next, a method for isolating the new bacterial strain of the present invention from soil will be described. The new bacterial strain of the present invention is isolated by conventional methods. That is, soil collected from Canberra, Australia was appropriately diluted with sterilized water prepared in advance, and then spread on a TA isolation agar medium with the composition shown below, and cultured at 28°C for 10 days. This strain can be isolated from among the colonies of actinomycetes. TA medium Trehalose 10g L-asparagine 1g K ₂ HPO ₄ 0.5g ISP trace salt solution 1g Nystatin 0.025g Agar 20g Distilled water 1000ml PH 7.0 The medium used to isolate the new bacterial species of the present invention includes a carbon source and nitrogen. Any synthetic or natural medium can be used as long as it contains appropriate amounts of nutrients selected from sources, inorganic ions, organic nutrients, and the like. Above, SANK62781, SANK62881,
Although we have explained the SANK62981 and SANK62585 strains, the properties of actinomycetes are not constant;
It is well known that the strain can easily change naturally or artificially, and the strain that can be used in the present invention belongs to the genus Nocardia or Streptomyces, and ML-
236B derivative as 3″,6′-dihydroxy-ML-236B
This includes all strains that can be converted into derivatives. When carrying out the method of the present invention, enzymatic hydroxylation can be carried out by culturing the converted bacteria under culture conditions suitable for its growth, and adding and contacting the raw material compound into the medium.Cultivating the converted bacteria. - A method in which bacteria are collected and the resulting converted bacterial cells are brought into contact with a raw material compound, and a method in which a cell-free extract prepared from the converted bacterial cells is brought into contact with a raw material compound are employed. The converting bacteria can be cultured in a medium containing nutrients that can be used by microorganisms. As the nutrient source, any known nutrient source used for general microbial culture can be used. For example, glucose, sucrose, starch, glycerin, starch syrup, molasses, soybean oil, etc. can be used as carbon sources. Further, as the nitrogen source, soybean flour, wheat germ, meat extract, peptone, corn steep liquor, dried yeast, ammonium sulfate, etc. can be used. In addition to other inorganic salts such as table salt, potassium chloride, calcium carbonate, and phosphates, additives necessary to support the growth of bacteria and promote the production of the enzymes having hydroxylation ability may be used in combination as appropriate. can. As a culture method, a culture method generally used for derivatives, such as a liquid culture method, is possible, and a deep culture method is suitable from an industrial perspective. Cultivation is carried out under aerobic conditions, with a culture temperature of 20~20°C.
37°C, preferably 26-35°C. The method is carried out by adding raw material compounds and culturing. The timing of addition varies depending on the optimal culture conditions of the converting bacteria used, especially the culture equipment, medium composition, culture temperature, etc., but it is best to add it when the hydroxylation ability of the converting bacteria begins to increase, and usually 1 day after the start of culturing of the converting bacteria. Preferably, after ~3 days have elapsed. The amount of the raw material compound added to the medium is selected from the range of 0.01 to 5.0%, preferably from 0.05 to 2.0%. Cultivation after addition of the raw material compound is carried out under aerobic conditions at the above-mentioned culture temperature. The culture period is 3 after the addition of the raw material compound.
~5 days. The method is to culture the converting bacteria in the presence of a small amount of substrate by the method described above, and culturing the converting bacteria until the hydroxylation ability of the converting bacteria reaches its maximum. In other words, the hydroxylation ability depends on the type of medium,
Although it varies depending on the temperature etc., it usually takes 4 to 4 hours after the start of culture.
Since it reaches its maximum level in 5 days, the culture is terminated at this point. Bacterial collection is performed by subjecting the culture to centrifugation, straining, etc. It is preferable that the collected converted bacterial cells be washed with normal saline, buffer solution, etc. before use. In order to bring the thus obtained converted bacterial cells into contact with the starting compound, it is usually carried out in an aqueous medium, e.g.
It is carried out in a phosphate buffer with a pH of 5-9. The reaction temperature is 20-45°C, preferably 25-35°C. The concentration of the raw material compound can usually be selected from a range of 0.01 to 5.0%.
The reaction time depends on the concentration of the raw material compound, the reaction temperature, etc., but is usually about 1 to 5 days. The cell-free extract in this method is obtained by applying physical or chemical means to the transformed bacterial cells obtained in the above method, such as by grinding, ultrasonication, etc. Obtained as a bacterial cell lysate by treatment with an activator or an enzyme. The method of contacting the cell-free extract thus obtained with the raw material compound is carried out in the same manner as the method of contacting the converted bacterial cells with the raw material compound described above. After the conversion reaction is completed, the target compound can be directly collected, separated, and purified from the product by known methods. For example, the product is filtered, the resulting filtrate is extracted with an organic solvent that is not easily miscible with water, such as ethyl acetate, and the solvent is distilled off from the extract. It can be separated and purified by subjecting it to column chromatography using, etc., and eluting with an appropriate eluent. Furthermore, if desired, the obtained product can be converted into the target compound by subjecting it to a hydrolysis reaction, salt formation reaction, esterification reaction, or lactonization reaction according to conventional chemical methods, and can be easily collected. can. All of these methods are conventional methods, such as the following method. Carboxylic acids with formula () are used when the product of the conversion reaction is a carboxylate salt, and the resulting liquid
It can be obtained by adjusting the pH to 4 or less, preferably 3 to 4. The acid used is not limited to organic acids or mineral acids as long as it does not affect the target compound; for example, trifluoroacetic acid, hydrochloric acid, sulfuric acid, etc. are preferably used. The carboxylic acid thus obtained is extracted,
After processing such as washing and dehydration, it can be used in the following reaction. A metal salt of a carboxylic acid having the formula () can be obtained by contacting a hydroxide, carbonate, etc. of the metal with the above carboxylic acid in an aqueous solvent. Examples of aqueous solvents used include water; methanol;
Alcohols such as ethanol, acetone, n
- A mixed solvent of water and an organic solvent such as hexane or ethyl acetate is suitable. Particularly suitable is a mixed solvent of a hydrophilic organic solvent and water. The reaction is usually suitably carried out near a room, but may be carried out under heating if necessary. Amine salts of carboxylic acids having formula () are obtained by contacting amines with the above carboxylic acids in an aqueous solvent. Examples of the aqueous solvent used include water; alcohols such as methanol and ethanol; ethers such as tetrahydrofuran; and mixed solvents of water and nitriles such as acetonitrile; preferred is aqueous acetone. The reaction is usually preferably carried out at a temperature of 7 to 8.5°C and below room temperature, particularly at a temperature of 5 to 10°C. The reaction is completed instantly. Alternatively, the carboxylic acid metal salt obtained above can be dissolved in an aqueous solvent, and then a mineral acid salt (eg, hydrochloride) of the desired amine can be added under the above conditions to obtain the salt exchange reaction. Amino acid salts of carboxylic acids having the formula () are:
It is obtained by contacting an amino acid with the above carboxylic acid in an aqueous solvent. Examples of the aqueous solvent used include water; a mixed solvent of water and alcohols such as methanol and ethanol; and ethers such as tetrahydrofuran. The reaction is usually carried out under heating, preferably around 50 to 60°C. The alkyl ester of a carboxylic acid having the formula () can be obtained by contacting the carboxylic acid obtained above with an alcohol. In this case, a mineral acid such as hydrochloric acid or sulfuric acid, boron fluoride, an acidic ion exchange resin, or the like is used as a catalyst, and as a solvent, the same alcohol or one that does not participate in the reaction, such as benzene, chloroform, or ether, is used. Alternatively, it can be obtained by contacting the carboxylic acid obtained above with a diazoalkane. The reaction is usually carried out by contacting with an ethereal solution of the diazoalkane. or,
It is obtained by contacting the metal salt of carboxylic acid obtained above with an alkyl halide. Suitable solvents to be used include, for example, dimethylformamide, tetrahydrofuran, dimethylsulfoxide, and acetone. All reactions are preferably carried out at around room temperature, but depending on the type of reaction system, they may be carried out under heating if necessary. The lactone form of carboxylic acid having the formula () is
It is obtained by contacting the carboxylic acid obtained above with a catalytic amount of acid. The acid used is preferably an organic or mineral acid such as trifluoroacetic acid, hydrochloric acid, or sulfuric acid. The reaction is usually suitably carried out at around room temperature. Furthermore, the target compound thus obtained can be used as a raw material and converted into other target compounds according to the above-mentioned conventional chemical methods. The target compound thus obtained can be collected, separated, and purified by appropriately combining various methods. For example, this can be carried out by adsorption or ion exchange chromatography using various carriers such as activated carbon or silica gel, Gegel filtration using a Sephadex column, or extraction using an organic solvent such as ether, ethyl acetate, or chloroform. In particular, separation of isomers can be carried out by the above separation and purification means at an appropriate time after the completion of the conversion reaction or after the completion of the desired step. Examples will be shown next, but the present invention is not limited thereto. Reference Example 1 Isolation of a new strain of Streptomyces carbophilus 1 g of soil and soil-attached plant pieces collected from Canberra, Australia were suspended in 9 ml of sterile water, thoroughly stirred with a mixer, and then left for about 30 minutes. Soil suspension supernatant 1 obtained in this way
ml was diluted 1000 times, and 0.05 ml of the supernatant was spread on an isolation agar medium TA having the composition shown below using a sterile Conlage rod. These agar media were cultured at 28°C for 10 days. Composition of agar medium TA for separation Trehalose 10g L-asparagine 1g K ₂ HPO ₄ 0.5g ISP trace salt solution 1g Nystatin 0.025g Agar 20g Distilled water 1000ml PH 7.0 Streptomyces carbophilus SANK62585 No. 1145 (FERM BP-1145)) was obtained by inoculating a yeast malt agar (ISP2) slant medium from the colonies that appeared on the TA agar medium prepared as above and culturing it at 28℃ for 14 days. It was hot. It was confirmed that this strain was a single strain by monocolony treatment. In addition, the pure cultured strain described above can be stored as a freeze-dried ampoule made using 10% skim milk as a dispersant. Example 1 3″α,6′β-dihydroxy-ML-236B carboxylic acid sodium salt (B substance) and 3″β,6′β-dihydroxy-ML-236B carboxylic acid sodium salt (A substance) and Nocardia sp. in 20 500 ml Erlenmeyer flasks containing 100 ml of culture medium with the following composition.
The SANK62981 strain was inoculated and cultured with shaking at 30°C and 220 rpm. After 2 days, ML-236B carboxylic acid sodium salt was added to a final concentration of 0.05%, and the culture was incubated at 26°C and 220 rpm for an additional 5 days. Cultured at pm. Medium composition: Glucose 1.0% Peptone 0.2 Meat extract 0.1% Yeast extract 0.1 Corn steep liquor 0.3 Tap water Remaining (PH uncorrected) After completion of the culture, the conversion reaction solution was filtered and the pH of the solution was adjusted to 8.5 with caustic soda. Next, the liquid was applied to a column packed with 600 ml of high porous polymer HP-20, washed with 3600 ml of water, and then 20% v/v
Elution was performed with 1200 ml of methanol. After concentrating the eluate under reduced pressure, lyophilize it to obtain 610 mg of solid material.
was gotten. The solid material obtained here was subjected to preparative liquid chromatography (column used: SSC-ODS).
-2942, φ30mm×250mmmm), 30%v/v
Elution was performed repeatedly using methanol as a developing solvent, and substance A was eluted first, and substance B was eluted next. The two substances thus obtained were each concentrated under reduced pressure and then lyophilized to obtain 50 mg of Substance A and 55 mg of Substance B. Of the obtained substances A and B, the hydroxyl group at the 3″ position of one is α-coordinated, and the other is β-coordinated.The physical property values of substances A and B are as follows. (1) Molecular formula: C ₂₃ H ₃₅ O _{8 Na} (2) Molecular weight: As a result of actual measurement by FAB-MS method, A
Both substance and substance B were 463 (M+H) ⁺ and 485 (M+Na) ⁺ . (3) Nuclear magnetic resonance spectrum: δ: ppm Nuclear magnetic resonance spectrum (270M) measured in heavy water using tetramethylsilane as an external standard.
Hz) as shown in FIG. 1 (material A) and FIG. 2 (material B). (4) High performance liquid chromatography: 4.833 minutes for substance A and 5.5 minutes for substance B under the following conditions.
It has a retention time of minutes. Column; ERC- with an inner diameter of approximately 6 mm and a length of approximately 10 cm.
ODS-1262 (manufactured by Elmer Optical Co., Ltd.) Mobile phase; methanol: water: glacial acetic acid: triethylamine (500:500:1:1) Column temperature: 40°C Flow rate: 1.0 ml/min Detector: UV238 nm (5) Gas chromatography: 24.563 minutes for substance B and 24.563 minutes for substance A under the following conditions.
It has a retention time of 25.475 minutes. Column; inner diameter approx. 0.2 mm, length approx. 25 m, film thickness
0.33 μm chemically bonded methyl silicone (manufactured by Heuret-Packard, ULTRA #1) Column temperature: Approximately 280°C Sample vaporization chamber and detector temperature: Approximately 300°C Carrier gas: Helium 2.0 ml/min Detector: Hydrogen flame Ionization detector example 2 3″α,6′β-dihydroxy-ML-236B carboxylic acid sodium salt (substance B) and 3″β,6′β-dihydroxy-ML-236B carboxylic acid sodium salt (substance A) below Streptomyces sp. in 20 500 ml Erlenmeyer flasks containing 100 ml of medium
Inoculated with SANK62585 strain, 27-28℃, 220r.pm
Cultured with shaking. Two days later, ML-236B carboxylic acid sodium salt was added to the final concentration of 0.05%, and the mixture was further cultured at 27-28°C and 220 rpm for 5 days. Medium composition Glucose 2.0% Peptone 1.0 Yeast extract 0.1 Tap water remainder (PH = 7.0) After completion of the culture, the conversion reaction solution was filtered and the solution was adjusted to PH = 8.5 with caustic soda. Next, the liquid was applied to a column packed with 600 ml of high porous polymer HP-20, washed with 3600 ml of water, and 20% v/v.
Elution was performed with 1200μ of methanol. After concentrating the eluate under reduced pressure, it was lyophilized to a solid mass of 720
mg was obtained. This solid was subjected to preparative liquid chromatography (column used: SSC-ODS-2942, φ30
mm x 250 mm) and repeated elution using 30% v/v methanol as a developing solvent, substance A was eluted first, and substance B was eluted next.
The two substances thus obtained were each concentrated under reduced pressure and lyophilized to obtain 45 mg of pure substance A and 38 mg of pure substance B. The physical property values of Substance A and Substance B were as described in Example 1. Test example Cholesterol synthesis inhibitory effect 3″,6′β-dihydroxy-ML consisting of a carboxylic acid having the above general formula (), a pharmacologically acceptable salt thereof, an ester thereof, or a closed ring lactone thereof
-236B derivative is 3-hydroxy-3-methylglutaryl coenzyme A reductase (3-hydroxy-3-methylglutaryl coenzyme A reductase), which is known as the rate-limiting enzyme in the cholesterol synthesis pathway.
-hydroxy-3-methyl-glutaryl-CoA
It was found that it specifically inhibits raductase.
Table 3 shows the cholesterol synthesis inhibitory effects of these compounds [measured by the method described in Journal of Biological Chemistry (J. Biol. Chem.) Vol. 234, p. 2335 (1959)]. 【table】

[Brief explanation of drawings]

FIG. 1 shows the nuclear magnetic resonance spectrum of substance A, and FIG. 2 shows the nuclear magnetic resonance spectrum of substance B.

Claims (1)

[Claims] 1 formula a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
3″,6′-dihydroxy-ML-236B derivative. 2 Formula a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
The compound according to claim 1, which is a 3″,6′β-dihydroxy-ML-236B derivative. a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
The compound according to claim 2, which is a 3″α,6′β-dihydroxy-ML-236B derivative. a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
The compound according to claim 2, which is a 3″β,6′B-dihydroxy-ML-236B derivative. a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
The ML-236B derivative is hydroxylated using a microorganism belonging to the genus Nocardia or Streptomyces to obtain the formula a pharmacologically acceptable salt thereof,
Consists of its ester or its ring-closed lactone
3″,6′-dihydroxy, characterized in that the 3″,6′-dihydroxy-ML-236B derivative is converted into a 3″,6′-dihydroxy-ML-236B derivative, and the 3″,6′-dihydroxy-ML-236B derivative is collected from a system containing the conversion reaction product. -Production method of ML-236B derivative. 6 In the production of 3″,6′-dihydroxy-ML-236B derivative, ML-
6. The production method according to claim 5, which comprises adding a 236B derivative and carrying out conversion culture. 7 In the production of 3″,6′-dihydroxy-ML-236B derivatives, Claim 5 comprises culturing and collecting converted bacteria, and bringing the obtained converted bacteria cells into contact with the ML-236B derivatives. The production method described in Section 8. In the production of the 3″,6′-dihydroxy-ML-236B derivative, the cell-free extract prepared from the converted bacterial cells is brought into contact with the ML-236B derivative and enzymatically hydroxylated. The manufacturing method according to claim 5. 9 The microorganism belonging to the genus Nocardia is Nocardia sp.
No.), SANK 62881 (Feikoken Bikyo No. 6182) or SANK 62981 (Feikoku Kenbiyori No. 6183) as stated in claim 5, 6, 7, or 8. manufacturing method. 10 Claim 5 in which the microorganism belonging to the genus Streptomyces is Streptomyces carbophilus SANK62585 (Feikoken Jokyo No. 1145)
The manufacturing method according to item 6, item 7, or item 8.