JPS62174040A - 3"-hydroxy-ml-234b derivative and production thereof - Google Patents

Abstract

NEW MATERIAL:A 3''-hydroxy-ML-2346B derivative consisting of a carboxylic acid having formula I, a pharmacologically acceptable salt, ester or ring-opened lactone derivative thereof. EXAMPLE:3''-Hydroxy-ML-236B carboxylic acid. USE:With inhibitory action on synthesis of cholesterol, capable of reducing lipid in blood, useful as, e.g. a remedy for hypertension and preventive agent against arteriosclerosis and orally or parenterally administered. PREPARATION:A microorganism belonging to the genus Streptomyces and capable of converting an ML236B derivative consisting of a carboxylic acid having formula II, salt, ester or ring-opened lactone derivative thereof into the 3''-hydroxy-ML-236B derivative expressed by formula I, e.g. Streptomyces sp. SANK62285 (FERM-P No. 1142), is cultivated in a culture medium containing the ML-236B derivative by a method for cultivation, etc., to afford the aimed compound expressed by formula I.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a 3''-hydroxy-ML-236B derivative consisting of a carboxylic acid having the formula, a pharmacologically acceptable salt thereof, an ester thereof or a closed ring lactone thereof, and a method for producing the same.

Conventionally, in the general formula (I), the hydroxy group at the 3-position is substituted with a hydrogen atom and the 7' (ML-236B derivatives are disclosed in, for example, JP-A-50-155690 and JP-A-53-5631).
No. 4, No. 53-84954, and
MB-530 in which the hydrogen atom at the 6' position is substituted with a methyl group
The B derivatives are described in US Pat. No. 4,376,863, and all of them are known to exhibit cholesterol synthesis inhibitory activity.

The present inventors discovered a carboxylic acid having the general formula (1)k,
The present invention was completed based on the discovery that a pharmacologically acceptable salt thereof, an ester thereof, or a closed ring lactone thereof exhibits an inhibitory effect on cholesterol synthesis.

Examples of the pharmacologically acceptable salts of the compound having the general formula (I) 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, and aluminum salts, iron salts, zinc salts, copper salts, nickel salts, and cobalt salts. Among these, alkali metal salts, alkaline earth metal salts and aluminum salts are preferred, and sodium salts, potassium salts, calcium salts and aluminum salts are most preferred. Preferred amino acid salts include basic amino acids such as arginine, linone, histidine, α, γ-diaminobutyric acid, and ornithine.

Examples of amine salts include t-octylamine, nobennolamine, nocyclohexylamine, morpholine, D-
Preferred are phenylglycine alkyl ester, D-glucosamine, and the like.

Examples of the ester of the compound having the general formula mk include alkyl esters such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and pentyl. Methyl is preferred.

The ring-closed lactone of the compound having the general formula (1) above refers to a compound in which formula (1) is represented by the following ring-closed structural formula.

Examples of the carboxylic acid having the general formula (Il), a pharmacologically acceptable salt thereof, an ester thereof, or a ring-closed lactone thereof obtained according to the present invention include the compounds fh described below.

1) 3〃-Hydroxy-ML-236B carboxylic acid 2)
3"-hydroxy-ML-236B carboxylic acid sodium salt 3) 3JI-hydroxy-ML-236B carboxylic acid potassium salt 4) 3"-hydroxy-ML-236B carboxylic acid calcium salt 5) 3"-hydroxy-ML-236B carvone acid aluminum salt 6) 3-Hydroxy-ML-236B carboxylic acid arginine salt 7) 3"-hydroquine-ML-236B carboxylic acid lysine salt 8) 3/L-hydroxy-ML-236, B carboxylic acid t-octylamine Salt 9) 3-Hydroxy-ML-236B carboxylic acid D-
Phenylglycine ethyl ester salt IQ) 3//
-hydroxy-ML-236B carboxylic acid methyl ester 11) 3"-hydroxy-ML-236B longevity i--1
Lactone Form In the general formula (1) of the present invention, various geometric isomers exist depending on the arrangement of substituents.

Furthermore, various optical isomers also exist due to the presence of asymmetric carbon atoms. In the general formula (1), these isomers and mixtures of these isomers are all represented by a single formula. Therefore, in the present invention, the general formula (I
), its pharmacologically acceptable salts, its esters, or its ring-closed lactones include not only these isomers but also mixtures of these isomers.

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 administered to adults in 3 to 4 divided doses of about 0.2 to 200 qi per day.

However, a larger amount can be used if necessary.

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 shown by the following formula and 3-hydroxy-3-methylglutaryl coenzyme A reductase, which is the rate-limiting enzyme for cholesterol biosynthesis in enzyme systems isolated from experimental animals and cultured cell systems. It is known to exhibit a strong serum-z-lowering effect even at the individual animal level (
JP-A-50-155690, Journal of Antibiotics, Vol. 29, pp. 1346-1348, 1976
Year).

The object compound of the present invention is ML-236, which consists of cardanic acid having the formula, its pharmacologically acceptable salt, its ester, or its ring-closed lactone.
The B derivative is enzymatically hydroxylated by contacting with a 3-hydroxyl converting bacterium or its cell-free extract, and then the resulting conversion reaction product is subjected to hydrolysis reaction, salt formation reaction,
It is obtained by collecting from one reaction solution subjected to esterification reaction or lactonization reaction.

Examples of microorganisms that can convert the raw material compound into cardanic acid having the general formula (1), a pharmacologically acceptable salt thereof, an ester thereof, or a closed ring lactone thereof include the genus Streptomyces.

Among the microorganisms belonging to the genus Streptomyces, especially -10- Streptomyces sp.
ssp, ) 5ANK 62285 (Feikokenchayo No. 11 Cow)) Streptomyces sp. (S, tre
ptomyssp, ) 5ANK 62385
(Fiber Engineering Research Institute No. 1+3) Streptomyces sp. 5AN
K 62485 (Feikoken Miyori No. 114+) is 90-
It has the ability to convert the ML-236B derivative into the 3"-hydroxy=ML-236B derivative at the above conversion rate.

The mycological properties of these strains are as follows.

■ Morphological characteristics The morphology is based on the ISP [International Streptomyces Project (InternationalStreptomyces Project)].
eptomyces Project)) After culturing on a specified medium at 28°C for 14 days, the cells were observed under an optical and electron microscope. 5ANK 62285.5ANK 62
In both strains 385 and 5-ANK 62485, the basal hyphae are branched and elongate well, and the aerial hyphae are simply branched.

The morphology of the spore chain of the 5ANK62285 strain is straight, the spores are spherical to oval, and the surface structure is smooth.

The morphology of the spore chains of the 5ANK 62385 and 5ANK 62485 strains is helical, the spores are spherical to oval, and their surface structure is smooth. 5ADIK 62285
．． Special organs such as axle branches of aerial hyphae, sclerotia, and sporangia were not observed in strains 5ANK62385 and 5ANK62485.

2. Various notes on various culture media The properties after culturing on various culture media at 28℃ for 14 days are as follows.
As shown in Table 3. The color tone display represents the color chip number of the Nippon Color Research Institute version, PA Standard Color Chart.

Table 1 Type of culture medium Item 5 Characteristics of ANK 62285 strain Sucrose G Not very good, yellowing method (
1-9-10) '61tM61℃ AM Good,
Powder, yellowish ash to light brownish ash (1-9-10 to l-7-
10) B Bright brownish ash (1-7-10) SP Produced white glucose G Good, yellowish method (1-9-10
)7-1 /R5q 7AM Slightly formed, grayish white (N-9) agar L Yellowish method (1-9-1
0) sp Not produced, glycerin G Not very bright, light pink (2
-8-4) 7, ni/e″″′″7ALA Slightly formed, gray white (N-9) agar (ISP 5),
Light pink (2-8-4) SP Light orange (3-
9-4) Starch/Inorganic salt G Good, gray tea (4-6-6) B
Brown (3-4-6) sp Light red matcha (3-7-4) Tyrosine agar G Good, brown (6-5-6) (I
SP 7) A, good, powdery, light brownish ash (1-8-1("'B brownish black (1-1-6) 5p brown (6-5-6) peptone/G good, light yellowish brown (4) -8
-9) R light yellow tea (4-8-9) sp No production, nutritional agar 0 Very good, light yellow (3-9-1
0) (Dlfoo) AM Slightly formed, grayish white (N
-9) R light yellow (3-9-10) SP Produced pear yeast extract・G Very good, yellowish brown (4-5
-8) R tea body black (1-1-6) sp brown (6-4-6) oatmeal agar (ISP 3) () Good, dark yellowish brown ()
3-4-8) AM Abundantly formed, powdery, brownish ash (2
-6-9) B Dark yellowish tea (2-3-7) sp Grayish tea (4-6-6) Potato extract/G Not good, brownish ash (3-5
-6) R dark brown (3-3-6) sp No production G: Growth, Tang: Aerial mycelium, B: Back side, SP: Soluble color 2nd surface Type of medium Item SAI, IK Properties of strain 62385 Nuucrose・G Not so good, tribute bed ash (1-
9-10) Nitya Shio Agar AM Good, powdery, light brownish ash (1-7-10) B Yellowish method (2-8-10
) SP does not produce crucose/G Not so good, yellowing method (1-
9-10) R yellowing method (1-9-10) sp produced zezuglycerin・G good, tribute ash (1-9-10
) B Yellowing method (2-9-10) sp Produced red starch/inorganic salt G Good, yellowing method (2-8-10)
sp Not derived Tyrosine agar G Very good, yellowish (10-
6-8) (I SP 7) A', □, A
Slightly Hikigata Yaku, pale yellow death Orenno (9-9-9) R yellowish type (
10-6-8) sp Production Zezuhapton・G Not very good, light yellowish tea (6
-8-9)E Light yellow tea (6-8-9) SP No production, nutritional agar G Not very good, light yellow tea (6-8
-9) (D"fco) ,, No formation R Light yellowish tea (6-8-9) sp Produced zezu yeast extract・G Good, yellowish (8-7-9)
) R Nibu yellow to grayish yellow tea (10-7-9 to 4-5-9)
Oatmeal agar G Very good, yellowish (s-
6-s)B Yellowish brown to brown (6-7-8 to 4-4-6)S
P Raw potato extract・G Not good, yellowing method (1-9
-10) B Yellowing method (1-9-10) sp Production Zezu Table 3 Type of medium Item 5 ANK, 62485 strain properties Sucrose G Not good, Yellowing method (1-9
-10) Nitrous salt agar A, slightly formed, gray mortar (
N-9) R yellowing method (1-9-10) sp No production, glucose G Not good, yellowing method (1-9
-10) B yellowing method (1-9-10) sp No glycerin production, G not so good, yellowing method (1-9-10)
1-9-10) R yellowing method (1-9-10) sp No production, starch/inorganic salts G Good, olive ash (3-7
-10) B Bright brownish gray (2-7-10) sp Produced Zezu Tyrosine Agar G Very good, yellowish (10-
6-4) (ISP 7) Store Good, powdery, light brown ash (2-8-7) B Bright brown (8-5-6) Covert G Not very good, light yellowish ash (6- 8-9) B Light yellow tea (6-8-9) sp Produced Zezu Nutrient Agar G Not very bright, light yellow tea (6-8-9)
6-8-9) (Difco) Store No formation R light yellowish tea (6-8-9) sp No formation yeast extract G Good, yellowish (8-6-
9) B Yellowish color (8-6-9) SP Produced red pepper oatmeal agar G Good, yellowish color (8-6-8
) 1'l Dark brown (4-4-8) Potato extract/G Not good, yellowish ash (1-9
-10) Old yellowish gray (1-9-10) sp production Zezu 3, physiological properties 5ANK62285.5ANK 62385 and 5A
The physiological properties of the NK 62485 strain are shown in Tables 4-6.

Table 4 Melting of curved starch of 5ANK 62285 strain Liquefaction of positive gelatin Reduction of positive nitrate Coagulation of negative milk 4-notonization of positive milk Positive growth temperature range (Medium 1)* 10-40℃ Suitable growth temperature (Medium 1) 20-30℃ salt tolerance (medium 1)
Grows up to 7 inches, does not grow at 10% and produces melanin-like pigment (Medium 2) Negative (Medium 3) Negative No. 5 Deicing of surface starch Liquefaction of positive gelatin Positive reduction of nitrate Coagulation of negative milk Peptone in positive milk [H] Positive growth temperature range (medium'D 10-40℃ suitable growth temperature (medium 1) 26-28℃ salt tolerance (medium 1)
Growth up to 7%, no growth at 10th stage, melanin-like pigment production @
Note (Medium 2) Negative (Medium 3) Negative (Medium 4) Negative Table 6 Properties of 5ANK 62485 Strain Starch thawing Liquefaction of positive gelatin Positive nitrate reduction Coagulation of negative milk Peptonization of positive milk Positive growth temperature range (medium 1) 10-40℃ suitable growth temperature (medium 1) 20-28℃ salt tolerance (medium 1)
Growth up to 7%, no growth at 10 cm Melanin-like pigment production (Medium 2) Negative (Medium 3) Negative (Medium 4) Negative*: Medium 1; Yeast extract: Malt extract agar (ISP
2) 2 Nidripton Yeast Extract Broth (ISP i
) 3; peptone yeast extract iron agar (ISP 6) 4; tyrosine agar (ISP 7)
The carbon source assimilation ability after 4 days of culture was investigated. 5ANK62
Since 285.5ANK62385 and S oil 62485 strains show some growth even in a control medium without carbon source addition, it is difficult to accurately describe their assimilation properties. For reference, Table 7 shows the relative assimilation ability when the control is -.

Table 7 +: Used, ±: Weakly used, -: Not used 4
Regarding bacterial components 5ANK62285.5ANK62385 and 5AN
The cell wall of K 62485 strain was prepared using the method of B. Pelaka et al.
B, Beckere tal −+ Applied・
Applied Microbiology
12, pp. 421-423.

1964], L, L-noaminopimelic acid and glycine were detected, so it was confirmed that the cell wall was of type II. Also, 5ANK62
285.5ANK62385 and 5ANK6248
Sugar components in whole cells of the five strains were analyzed using the M.P. Lechevalier method (M, P. Lechevalier, Journal of Laboratory Medicine).
and Clinical Medicine),
71, p. 934, 1968], no characteristic pattern was observed.

From the above, it was determined that these three strains belong to the genus Streptomyces among actinomycetes, and therefore they are considered to be Streptomyces sp.
, ) 5ANK 62285.5ANK 62385
and 5ANK 62485.

The 5ANK62285.5ANK62385 and 5ANK62485 strains were identified by ISP (So International Streptomyces pronoect.
he International Streptom
yces Project)) Standard, Parnose
Bergey's Manual ofD
stsrminative Bacteriology
) 8th edition, No Actinomycetes (The Act
inomycetes) Volume 2 and recent literature on actinobacteria.

The 8ANK62285.5ANK62385 and 5ANK62485 strains have been described above, but it is well known that the properties of actinomycetes are not constant and can easily change naturally or artificially, and the strains that can be used in the present invention belongs to the genus Streptomyces and includes all strains capable of converting ML-236B derivatives into 3"-hydroxy-ML-236B derivatives.

When carrying out the method of the present invention, enzymatic hydroxylation can be carried out by culturing the converting bacteria under culture conditions suitable for its growth, and (1) adding the raw material compound to the medium and bringing it into contact with the converting bacterium. The following methods are adopted: (1) cultivating and collecting the converted bacterial cells, and contacting the resulting converted bacterial cells with the raw material compound; and (2) bringing a cell-free extract prepared from the converted bacterial cells into contact with the raw material compound.

-2.1- Conversion 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. Nitrogen sources include soybean flour, wheat germ, meat extract,
Peptone, corn steep liquor, dried yeast, ammonium sulfate, etc. may 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 microorganisms, such as a liquid culture method, can be used, and a deep culture method is suitable for industrial use.

Cultivation is carried out under aerobic conditions, with a culture temperature of 20-37°C.
, preferably 26-28°C.

Method (2) is carried out by adding raw material compounds and culturing. The timing of addition depends on the optimal culture conditions of the conversion bacteria used,
It varies depending on the culture equipment, medium composition, medium temperature, etc.
The best time is when the hydroxylation ability of the converting bacteria begins to increase, and usually 2 to 3 days after the start of culturing the converting bacteria.

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 0.5%. 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 to 5 days after addition of the raw material compound. In the method (2), the converted bacteria are cultured in the presence of a small amount of substrate according to the method described above, and the bacteria are cultured until the hydroxylation ability of the converted bacteria reaches its maximum. In other words, the hydroxylation ability depends on the type of medium,
Although it varies depending on the temperature, etc., the maximum value is usually reached 4 to 5 days after the start of the culture, so the culture is terminated at this point. Bacterial collection is performed by subjecting the culture to methods such as centrifugation and filtration. It is preferable that the collected converted bacterial cells be washed with normal saline, buffer solution, etc. before use.

The thus obtained converted bacterial cells are brought into contact with the raw material compound, usually in an aqueous medium, for example in a phosphate buffer solution with a pH of 5 to 9.

The reaction temperature is 20-45°C, preferably 25-30°C. The concentration of the raw material compound is usually selected from the range of 0.001 to 50%. 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 obtained by the above method is obtained by applying physical or chemical means to the transformed bacterial cells obtained by the above method, for example, by grinding, ultrasonication, etc., to obtain a cell-free extract. Obtained as a bacterial cell lysate by treatment with surfactants, enzymes, etc.

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 ffi is filtered, the obtained solution E 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 a solvent, etc., and eluting with an appropriate bath separation agent.

Furthermore, the obtained product is converted into the target compound by subjecting it to a hydrolysis reaction, a salt-forming reaction, an esterification reaction, or a lactonization reaction according to a conventional chemical method, if desired.
It can be easily collected.

All of these methods are conventional methods, such as the following method.

When the product of the conversion reaction is a carboxylic acid salt, the carboxylic acid having formula (1)'r can be used to
Preferably, it is obtained by adjusting the pH to 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 can be used in the following reactions after being subjected to treatments such as extraction, washing, and dehydration.

The metal salt of cardanic acid having the formula mk can be obtained by bringing a hydroxide, carbonate, etc. of the metal into contact with the above carboxylic acid in an aqueous solvent. Suitable examples of the aqueous solvent used include water; a mixed solvent of water and alcohols such as methanol and ethanol, and organic solvents such as acetone, n-hexane and ethyl acetate. Particularly suitable is a mixed solvent of a hydrophilic organic solvent and water. The reaction is usually carried out at around room temperature, but may be carried out under heating if necessary.

The amine salt of the carboxylic acid at m' is obtained by contacting the amine with the carboxylic acid in an aqueous solvent. Examples of aqueous solvents that can be used include water; methanol, alcohols such as ethanol/lunato, ethers such as tetrahydrofuran, mixed solvents of nitriles such as acetonitrile, and water; however, hydrous acetone is preferable. be. The reaction is usually suitably carried out at pH 7-85 and below room temperature, particularly at 5-10°C. The reaction is completed instantly. Alternatively, the carbic acid metal salt obtained above may be dissolved in an aqueous medium, and then a mineral salt of the desired amine (such as hydrochloride) may be added under the above conditions to undergo a salt exchange reaction. can.

The amino acid salt of carmenic acid having formula (1) can be obtained by contacting the 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 at a temperature of 50 to 6
It is carried out at around 0°C.

The alkyl ester of a carboxylic acid having the formula (1) 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, one that does not participate in the reaction, such as the same alcohol or benzene, chloroform, or ether, is used. Alternatively, it can be obtained by contacting the carboxylic acid obtained above with a noasialkane. The reaction is usually carried out by contacting the noasialkane with an ether solution. Alternatively, it can be 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, trimethylsulfoxide, 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 represented by formula (11') can be obtained by contacting the carboxylic acid obtained above with a catalytic amount of acid. Examples of the acid used include trifluoroacetic acid, hydrochloric acid, and sulfuric acid. Organic acids or mineral acids are preferred.The reaction is usually suitably carried out 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 and separated by a suitable combination of various methods. , can be purified. For example, this can be carried out by adsorption using various carriers such as activated carbon or silica gel, ion exchange chromatography, gel filtration using a Sephadex column, extraction using an organic solvent such as ether, ethyl acetate, chloroform, etc.

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.

Example 1 3"-hydroxy-ML-236B lactone glucose 20, peptone 10, W mother extract 1.0 (f/A)
, 100 mf of medium with pH 7.0! 5 containing
Streptomyces sp. 5ANK62285 strain was inoculated into four Erlenmeyer flasks each having a capacity of 0.00 ml, and cultured with shaking at 28°C for 48 hours (22 Or, p, m).

The obtained seed culture solution 150- was transformed into a medium 15 having the same composition as above.
! 30 including! They were transferred to 2 units of Gloss-7 Armenter, and 0.01q6 of CB-442 was added as an antifoaming agent, resulting in an aeration volume of 15! /min, internal pressure 10jyA"TrL2, stirring speed 100-200r, p, m, and culture temperature 28°C for 45 hours. ML-236B calzenate sodium salt was added to this to a final concentration of 0.1%. After culturing was continued for another 50 hours, the culture solution was completely filtered to obtain p solution.

15! Gloss - P liquid 26 for 2 units! a) IP-20 resin 2
! After adsorbing with sv4~5, wash with water (8k), 30
Thiacetone-water 3! , 50 thiacetone - 8 parts water. 50 thiacetone one water bath fraction 8! was concentrated under reduced pressure to remove acetone. Concentrate 46! with 6N-hydrochloric acid
After adjusting H2 to 8 to 30, add an equal amount of ethyl acetate (4Ax2
) was extracted. This ethyl acetate extract was mixed with saturated saline (4
After washing with water with AX2), it was dried over anhydrous sodium sulfate and concentrated to obtain an oily concentrate 201.

The obtained oily concentrate 201 was dried with ethyl acetate 100
-, 0.5 mt of gold in trifluoroacetic acid was added thereto, and the mixture was allowed to stand at 50°C for 3 hours to perform lactonization. The reaction solution was diluted with 400% ethyl acetate, treated with 1% bicarbonate solution, washed with saturated brine, and diluted with anhydrous sulfuric acid.
It was dried with Jum. Concentrate this ethyl acetate solution (lOM)
128 grams of precipitated crude crystals of 3''-hydroxy-ML-236B lactone were separated from each other. A portion of these crude crystals was purified by silica gel column chromatography (si 60 ).

The pure 3"-hydroxy-ML-236B lactone compound prepared with n-hexane-acetone (0 to 50 v/v%) exhibits the following physical constants.

1) Molecular formula “23” 3406 2) Molecular weight: 406 (SIMS mass spectrometry spectrum:
QM-+41,407) 3) Infrared absorption spectrum The infrared absorption spectrum measured in Niji Box-1aX KBr is as shown in FIG.

4) Carbon-13-nuclear magnetic resonance vector: δ: ppm
Carbon-13-nuclear magnetic resonance spectrum (2
2,5MEz) is the following chemical shift) (ppm
) and multiplicity.

Chemical shift multiplicity 1 10.9 Q2
13.8 Q3 2
0, OQ 4 20.9 T5
23.6 T6 2
6.2 T7 30.9
D8 32.7
T9 36.9
Dlo 36.0 T1
1 37.6 D12
38.6 T13
45.9 DI4 62
．． 5 D15 68, L
D16 68.4
D17 76.2
DI8 123.6D
19 128.1 D20
132.8 D21
133.7 S22 17
0.8 S23 175.7
S Example 2 3"-hydroxy-ML-236B carboxylic acid sodium salt 3"-hydroxy-ML-25 obtained in Example 1
2.0 l of 6B lactone was dissolved in 200 methanol, and 50 m of distilled water was added thereto. Add approximately equimolar amount of sodium hydroxide solution C0.25f/40mZ H2 to this solution.
O) was fully applied at 40 to 50°C. During this time, the production of IJ salt was confirmed by liquid chromatography. Approximately 2 hours after the reaction was completed, the reaction solution was adjusted to pH 8.5 with N-hydrochloric acid, and then concentrated under reduced pressure (5o*: B, methanol was removed, and the aqueous layer was freeze-dried to give 3"-hydroxy-ML-236B. Carboxylic acid sodium salt 2.61 was obtained. 3"-Hydroxy-M L -236B carboxylic acid sodium salt exhibits the following physical constants.

l) Molecular formula: C25H3507・Na 2) Infrared absorption spectrum NijimaxcTL-1KBr
The infrared absorption spectrum measured inside is as shown in FIG.

3) Proton-nuclear magnetic resonance spectrum: δ: The proton-nuclear magnetic resonance spectrum (270 MHz) measured in ppm heavy water using tetramethylsilane as an internal standard is shown in FIG.

4) Carbon-13-nuclear magnetic resonance spectrum δ: ppm
Carbon-13-nuclear magnetic resonance spectra (225MH2) measured in heavy water using tetramethylsilane as an internal standard.
)l′! ,, the following chemical shifts (ppm) Chemical shift multiplicity 1 13.8 C214,
7C 321, 7C 421, 9D 5 25.2 T6
27.0 D7 31.
8 D8 35.5
T9 38.2
T10 38.3 T
11 44.6 T12
45.6 T13
486 D14 68.8
D15 69.4
D16 70.0
D17 711 D18
124, OD 19, 129.0 D20
134.5 D21
135.0 S22 1
77.7 S23 180.
9 S-4〇1 Example 3 Streptomyces sp. 5ANK 6 instead of Streptomyces sp. 5ANK62285 strain
Using the 2385 strain and the 5ANK 62485 strain,
The same procedure as in Examples 1 and 2 was carried out to obtain 3"-hydroxy-ML-236B lactone and 3"-hydroxy-ML-236B carboxylic acid sodium salt.

The physical constants were the same as those obtained in Examples 1 and 2.

Test Example Cholesterol Synthesis Inhibitory Effect The 3″-hydroxy-ML-236B derivative consisting of a carboxylic acid having the general formula (1) k, its pharmacologically acceptable salt, its ester, or its ring-closed lactone is rate-limiting on the cholesterol synthesis pathway. 3-hydroxy-3-methylglutaryl coenzyme A reductase (3-hydroxy-3-methylglutaryl coenzyme A reductase) is known as an enzyme.
-glutaryl-Co A reductase
) was found to specifically inhibit Cholesterol synthesis inhibitory effect of these compounds [Journal of Biological Chemistry (J, Biol.

Cne+n, ) Vol. 234, p. 2835 (1959)] are shown in Table 8.

Table 8 To the concentration μf that inhibits cholesterol synthesis by 50% 3″-Hydroxy-ML-236B Lactone body 00393″-Hydroxy-ML-236B Calse/acid sodium salt 0.0032M
L-236B lactone body (control) 0.010

[Brief explanation of drawings]

Figure 1 shows the infrared absorption spectrum of 3"-hydroxy-ML-236B lactone, Figure 2 shows the infrared absorption spectrum of 3"-hydroxy-ML-236B carboxylic acid sodium salt, and Figure 3 shows the same substance. The proton-nuclear magnetic resonance shear tor of is shown. Figure 3 6 5 tl 3
Two
PP'γn-11I-

Claims (1)

[Claims] 1. A 3″-hydroxy-ML-236B derivative consisting of a carboxylic acid having the formula ▲ Numerical formula, chemical formula, table, etc. ▼, a pharmacologically acceptable salt thereof, an ester thereof, or a closed ring lactone thereof 2. ML-236 consisting of a carboxylic acid having the formula ▲ Numerical formula, chemical formula, table, etc. ▼, its pharmacologically acceptable salt, its ester, or its ring-closed lactone form.
The B derivative can be converted into a 3″-hydroxy-ML-236B derivative consisting of a carboxylic acid having the formula ▲ Numerical formula, chemical formula, table, etc. ▼, a pharmacologically acceptable salt thereof, an ester thereof, or a closed ring lactone thereof. A microorganism belonging to the genus Streptomyces is cultured in a medium containing an ML-236B derivative, or an enzyme extract of the microorganism is brought into contact with an ML-236B derivative.
6B derivative is converted into a 3"-hydroxy-ML-236B derivative, and the 3"-hydroxy-ML-236B derivative is collected from a system containing the conversion reaction product. Manufacturing method.