JPS6320215B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing MB-530B carboxylic acid ester, a new physiologically active substance that has cholesterol biosynthesis inhibitory effects, anti-arteriosclerosis effects, anti-hyperlipidemic effects, etc. It is known that one of the causes of arteriosclerosis, hyperlipidemia, etc. is the accumulation of cholesterol in the body. Based on the idea that these diseases can be prevented or treated by inhibiting cholesterol biosynthesis, the present inventors systematically searched for cholesterol biosynthesis inhibitors in microbial culture solutions. Then, several inhibitory substances were collected from the culture fluid of Monascus sp. Through animal experiments using rats, this substance was found to have the effect of lowering the concentration of cholesterol in the blood and liver.
-530B and MB-530B carboxylic acid metal salts. MB-530B and MB-530B carboxylic acid metal salts are new substances and have the following chemical structures. (In the formula, M represents a metal element, and n represents the valence of the metal element.) As a result of further research, the present inventors found that the
We have discovered that MB-530B or MB-530B carboxylic acid ester derived from MB-530B carboxylic acid metal salt also has excellent cholesterol synthesis inhibitory effects, anti-arteriosclerosis effects, anti-hyperlipidemia effects, etc., and have developed the present invention. completed. MB-530B carboxylic acid ester has the following chemical formula. In the above formula, R is an alkyl group such as methyl, ethyl, n-propyl, isopropyl, n-butyl, n-hexyl; benzyl, 2-methylbenzyl, 3-methylbenzyl, 4-methylbenzyl,
2-ethylbenzyl, 3-ethylbenzyl, 4-
Ethylbenzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2-ethoxybenzyl, 3-ethoxybenzyl, 4-ethoxybenzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2- unsubstituted or substituted benzyl groups with alkyl groups, alkoxy groups or halogen atoms, such as bromobenzyl, 3-bromobenzyl, 4-bromobenzyl;
shows. MB-530B carboxylic acid ester represented by the above formula () is a new compound. MB-530B is collected from the culture by culturing microorganisms, and then MB
-Can produce 530B carboxylic acid ester. The microorganism that can be used in the present invention is an MB-530B producing bacterium belonging to the genus Monascus, preferably Monascus anka.
SANK10171 (IFO6540), Monascus pa-xii, SANK11172 (IFO8201),
Monascus purples
purpurous) SANK10271 (IFO4513), Monascus ruber (Monascus ruber) SANK11272
(IFO9203), SANK17075 (CBS832.70), same
SANK17175 (CBS503.70), SANK17275
(ATCC18199), SANK15177 (Microtechnical Research Institute
1340; FERM BP-1340), same SANK13778 (FERM BP-1343), same
SANK10671 (FERM BP No. 1342; FERM BP
-1342), SANK18174 (Feikoken Joyori No. 1342;
FERM BP-1342), Monascus vitreus SANK10960 (National Health Examination 609,
e-609) (FERM BP-
1344) is used. All of these bacteria have been deposited with the Institute of Microbial Technology, Agency of Industrial Science and Technology, Ministry of International Trade and Industry, or can be easily obtained from IFO, CBS, or ATCC. Among the above-mentioned producing bacteria, strains that the present inventors recognize as particularly effective include, for example, Monascus ruber.
SANK11272 (IFO9203) or four strains of Monascus ruber isolated and identified from soil by the present inventors, SANK15177, SANK13778,
They are the SANK18174 and SANK10671 strains, and the mycological characteristics of these four strains are as described below. SANK155177 bacterium: This bacterium is a strain isolated from the soil of Tsukimino, Yamato City, Kanagawa Prefecture. It grows well on potato dextrose agar medium at 25°C, produces yellowish brown to reddish brown soluble pigment in the culture medium, and produces basal hyphae. Many ascocarps are formed on the layer. On oatmeal agar medium, it becomes light brown and grows well, with good ascarp formation. The asci are formed on short stalks, spherical, 30-60 μm in diameter, and the stalks are almost colorless and branched, 25-60 x 3.5-5.0 μm. The asci are disappearing and are not clearly observed. Ascospores colorless, oval, smooth surface, 4.5-6.5
×4.0~5.0μm. Conidia are histosegmental, basophilically linked, 7.0-10.0 x 6.0-10.0 μm. Although this bacterium can grow at 37°C, the optimum temperature is around 23-30°C. SANK13778 bacterium: This bacterium is a strain isolated from the soil of Nagata, Inawashiro-cho, Yama-gun, Fukushima Prefecture. Its growth condition on the medium is similar to the above-mentioned strain, but it also grows pale reddish brown to reddish brown on potato dextrose agar medium and oatmeal agar medium. They differ in that they produce soluble pigments. The ascus is 35-75 μm in diameter, and the stalk is 30-70 x 3.5
5.0μm. Ascus is not observed. Ascospores colorless, oval, smooth, 4.5-6.0 x 4.0
~5.0μm. Conidia are 7.0-10.0 x 6.0-10.0 μm. SANK18174: This bacterium was isolated from the soil of Shakotan-cho, Shakotan-gun, Goshi Subprefecture, Hokkaido, and its growth condition on culture media is similar to the above-mentioned strains, but it produces a pale pink pigment on the above two types of culture media. The ascus is 20-70 μm in diameter, and the stalk is 20-60 x 3.0
5.0μm, no ascus observed. Ascospores colorless, oval, smooth, 5.0-7.0 x 4.0
~5.5μm. Conidia are chained, colorless, and often pear-shaped
6.0~9.5×6.0~10.0μm. SANK10671: This bacterium is a strain isolated from soil in Shinagawa Ward, Tokyo, and grows well on the two types of culture media mentioned above, and its condition is similar to the above strains, but the soluble pigment is dark red. The ascus is 30-80 μm, the stalk is 30-70 x 3.0
5.0μm, no ascus observed. Ascospores are colorless, ovoid, 4.5-6.5 x 4.0-
5.0μm. Conidia are colorless, pear-shaped to oval, 6.0-
10.0×6.0~8.5μm. From the above results, these four strains,
SANK15177, SANK13778, SANK18174, and SANK10671 were identified as Monascus ruber van Tieghem. The detailed mycological description of Monascus ruber is reported in the following literature. Masaki Takada, “Japanese Mycological Journal Materials (7)” (Bulletin of the Japanese Society of Mycology, Vol. 9, pp. 125-130, 1969) and its conidial generation, GTCole and WBKend-rick.
"Conidium ontogeny in hyphomycetes.The
imperfect state of Monascus ruber and its
Canadian Journal
of Botany, vol. 46, p. 987, 1968). It goes without saying that bacteria of the genus Monascus other than those mentioned above can also be used, regardless of their variants or mutant strains, as long as they exhibit the ability to produce MB-530B. MB-530B is produced in a culture by aerobically cultivating bacterial strains that produce these substances using a culture method known as a mold culture method. for example
The MB-530B production strain is subcultured on a potato-dextrose-agar medium manufactured by Difco, and in order to produce the substance, the cells grown on this agar medium can be directly inoculated into the production medium and cultured. Furthermore, the bacteria grown on the production medium can be cultured on a new production medium, and MB-530B can be produced there. MB-530B producing bacteria grows at 7-35℃,
A temperature of 20 to 30°C is usually preferred for production of the substance. M.B.
In order to cultivate Monascus bacteria that produce -530B, nutrients known for culturing molds and other microorganisms can be used. For example, glucose, maltose, dextrin, starch, lactose, sutucarose, glycerin, etc. can be used as carbon sources. Among these carbon sources, glucose and glycerin are the preferred carbon sources for the production of the substance. To produce MB-530B, all known nitrogen sources for mold and other microbial growth can be used. For example, peptone, meat extract, yeast, yeast extract, soybean flour, peanut flour, corn steep liquor, rice bran, inorganic nitrogen sources, etc. can be used. If necessary, inorganic salts, metal salts, and trace amounts of heavy metals can also be added. MB-530B is obtained by aerobically cultivating its producing bacteria, but it can be obtained using commonly used aerobic culture methods, such as
A solid state culture method, a shaking culture method, and an aerated agitation culture method are used. When antifoaming is required during culture or medium sterilization, antifoaming agents such as silicone oil and surfactants can be used. The culture temperature is preferably 20 to 30°C. The culture was continued until MB-530B was substantially accumulated, and the substance was extracted from the culture medium based on the properties of the substance revealed by the inventors, as shown in the Examples below. , can be carried out by appropriately combining various methods. That is, for example, ether, ethyl acetate,
Extract with a hydrophobic solvent such as chloroform, extract the bacterial cells with a hydrophilic solvent such as acetone or alcohol, and after concentration, dissolve in a highly polar solvent such as acetone or alcohol, or with a less polar solvent such as petroleum ether or hexane. Removal of impurities by chemical conversion, gel filtration using a Sephadex column, adsorption chromatography using activated carbon, silica gel, etc., high performance liquid chromatography, methods of purifying after making it less polar, and purification as a metal salt. Appropriately combine methods such as
Isolate MB-530B from the culture. MB-530B carboxylic acid metal salt is obtained by reacting MB-530B with a compound that forms a metal salt. In addition, MB-530B carboxylic acid metal salt is easily converted to less polar MB-530B in the presence of an acid catalyst. It also returns to metal salt under alkalinity such as caustic alkali or carbonate alkali. This mutual conversion can be performed quantitatively. MB-530B carboxylic acid 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. It will be done. MB-530B thus obtained can be reacted with various aliphatic or aromatic alcohols in the presence of acetyl chloride to produce MB-530B carboxylic acid esters. Also MB-530B
The carboxylic acid metal salt can be reacted with an alkyl halide or a benzyl halide, such as an alkyl iodide, to produce the MB-530B carboxylic acid ester. This will be explained in detail below using examples. Example 1 MB-530B carboxylic acid methyl ester glucose 5%, corn stave liquor
Medium containing 0.5%, peptone (Far East), 2% ammonium chloride (PH5.5 before sterilization) 300 to 600
Monascus louver in a culture tank
Inoculated with SANK18174, 26℃, aeration rate 300/min,
Cultivate for 116 hours with stirring at 190 revolutions/min, adjust the culture solution (including bacterial cells) to pH 3.4 with 6N hydrochloric acid, extract with methanol 800, add 6 kg of Hyflo Super Cell, filter through a filter press, and extract the methanol extract. Got 1100. Then, add this extract to saturated saline solution.
After washing with 200 and ethylcyclohexane 180, inhibitors were extracted with ethylene dichloride 600, and 50 g of trifluoroacetic acid was added to this extract and reacted at 80°C for 30 minutes. Add this extract to 2%
It was washed with 200 g of sodium bicarbonate solution and then with 200 g of 10% brine and concentrated to obtain 135 g of an oily substance. Dissolve this oil in 400 ml of methanol, of which 20 ml (approx.
6.8 g was applied to a high-performance liquid chromatography system 500 model (manufactured by Waters) for large-scale preparative separation equipped with a Prepack C ₁₈ column (reversed phase column). Developing solvent is methanol/water (85:15, v/v)
was developed at a flow rate of 200 ml/min (development time approximately 10 minutes) while observing with a differential refractometer connected to the main body, and the main peak indicated by the differential refractometer was fractionated. By repeating this operation, the main peak fractions obtained were collected and concentrated to obtain 10.2 g of an oily substance, which was dissolved in 30 ml of methanol.
ml (approximately 2 g as oil) was again subjected to the chromatography described above. Methanol/water (80:20, v/v) was used as the developing solvent at a flow rate of 200 ml/min, and the main peak portion was collected. This operation was repeated to collect and concentrate the main peak fractions to obtain 1170 mg of crude crystals from aqueous methanol. This crude crystal was further recrystallized with ethanol, and 864mg of
I got MB-530B. 200 mg of MB-530B was dissolved in 20 ml of methanol dehydrated with molecular sieve 3A, and several drops of acetyl chloride were added thereto, followed by stirring at room temperature for 3 hours. The solvent was distilled off, and 50 ml of ethyl acetate was added for extraction.
The extract was washed with 2% aqueous sodium bicarbonate and saturated brine, dehydrated with Glauber's salt, and the solvent was distilled off. Silica gel prepared by drying with benzene (Wako Gel C-
The fractions eluted with a developing solvent of ethyl acetate-benzene (6:94) were collected and the solvent was distilled off to obtain 65 mg of MB-530B carboxylic acid methyl ester as a colorless oil. Molecular weight 436.6 (mass spectrometry) Molecular formula C ₂₅ H ₄₀ O ₆ Specific optical rotation [α] _D = +208° (C = 1.05) Infrared absorption spectrum (CHCl ₃ ): Figure 1 Nuclear magnetic resonance spectrum (CDCl ₃ ): Figure 1 Figure 2 Example 2 MB-530B carboxylic acid methyl ester A medium containing 1.5% soluble starch, 1.5% glycerin, 2% fishmeal, and 0.2% calcium carbonate (before sterilization)
PH7.4) 300 was placed in a 600 volume culture tank, Monascus ruber SANK17075 (CBS832.70) was inoculated and cultured for 120 hours at a temperature of 26℃, aeration rate of 300/min, and stirring at 190 rpm. The liquid was applied to a filter press to obtain 35 kg of wet bacterial cells. Add 100% water to this and PH with caustic soda while stirring.
12 and left at room temperature for 1 hour, 2 kg of High Flow Super Cell was added and passed through a filter press. The solution was returned to pH 10 with hydrochloric acid, adsorbed on a column packed with HP-20 resin 5, and mixed with 15 parts water and 10
After washing with 90% methanol water, elution was carried out with 90% methanol. About 10% of the eluate was concentrated, and the remaining aqueous layer was adjusted to pH 2 with hydrochloric acid, and then extracted with ethyl acetate. The extract was washed with saturated saline, dehydrated with Glauber's salt, and concentrated to dryness, resulting in an oily substance containing MB-530B carboxylic acid.
Got 50g. Methanol was added to this to make 100 ml, and 20 ml of this was injected into a high-performance liquid chromatography system (same as that used in Example 1) for large-scale preparative separation equipped with a reverse phase column. Separate with 20% methanol water containing 2% acetic acid at a flow rate of 200 ml/min, and detect the main peak shown by a differential refractometer (7 to 10 minutes).
It was fractionated. This operation was repeated for the remaining 80 ml. After concentrating the peak fraction and removing methanol, it was extracted with ethyl acetate and concentrated to dryness while adding n-heptane to obtain 1.2 g of an oily substance. Furthermore, this was dissolved in 20 ml of methanol, and the above-mentioned chromatography was performed again under the same conditions.
- Obtained 150mg of 530B. To make the sodium salt, add 2 ml of methanol and 100 ml of water to 1N
Adjust the pH to 8.0 with caustic soda to make a clear aqueous solution, then adsorb it on a column filled with 10ml of HP-20.
After washing with 100ml of water, elute with 80% methanol,
After removing methanol by concentration, lyophilize to 130 mg.
A white powder of MB-530B carboxylic acid sodium salt was obtained. Here, the physical properties of MB-530B carboxylic acid sodium salt are as follows. 1 Elemental analysis value (%) Calculated value C; 64.84 H; 8.38 Na; 5.17 Actual value C; 64.78 H; 8.55 Na; 5.21 2 Molecular weight [Mass spectrometry (FDmass)] 444 3 Molecular formula C ₂₄ H ₃₇ O ₆ Na 4 Purple External absorption spectrum λ _nax 232nm (log ε=4.30) 239nm (log ε=4.36) 248nm (log ε=4.19) 5 Infrared absorption spectrum λ _nax cm ^-1 : Measured with KBr pellets. 3400, 1725, 1580, 1400 6 NMR spectrum (D ₂ O, δ: ppm) 0.85 (6H, multiplet), 1.1 (6H, multiplet), 3.7
(1H, multiplet), 4.1 (1H, multiplet), 5.35
(1H, Broadow multiplet), 5.6 (1H, Broadow multiplet), 6.0 (2H, multiplet) 7 High performance liquid chromatography Retention time 8.5 minutes (Column, Microbond Pack C ₁₈ , 60%
Methanol/water + PIC-A (Waters)
0.1%, flow rate 1.5ml/min) 8 Solubility Soluble in water, insoluble in organic solvents 9 Specific rotation [α] ²⁵ _D = +266° (C = 0.33, water) 100mg of MB-530B carboxylic acid sodium salt 2
ml of dimethyl sulfoxide, 50 μl of methyl iodide was added, and reaction was carried out at 40-50° C. for 5 hours while stirring with a reflux condenser attached. 5 ml of water was added to this reaction solution, extracted with 10 ml of dichloromethane, and the solvent was distilled off. The dried product was adsorbed on a silica gel column (Wako Gel C-100, 10 g) prepared with benzene, the fractions eluted with a developing solvent of ethyl acetate-benzene (4:96) were collected, and the solvent was distilled off to form an oily product.
35 mg of MB-530B carboxylic acid methyl ester was obtained. The physical properties of the obtained MB-530B carboxylic acid methyl ester were the same as in Example 1. As described above, MB-530B carboxylic acid ester can be produced by the method of Example 1 or Example 2, but below, various MB-530B carboxylic acid esters were prepared according to the method of Example 1. An example of the manufacturing method will be shown. Note that the manufacturing method of MB-530B is the same as in Example 1. Example 3 MB-530B carboxylic acid ethyl ester 100 mg of MB-530B was dissolved in 15 ml of ethanol dehydrated with molecular sieve 3A, and several drops of acetyl chloride were added thereto, followed by stirring at room temperature for 3 hours.
The solvent was distilled off, and 50 ml of ethyl acetate was added for extraction. The extract was washed with 2% aqueous sodium bicarbonate and saturated brine, dried over sodium sulfate, and the solvent was distilled off. Silica gel prepared by drying with benzene (Wakogel C-100, 10
g) Adsorbed with ethyl acetate-benzene (6:94)
Collect the fractions eluted with the developing solvent and distill off the solvent.
30 mg of MB-530B carboxylic acid ethyl ester was obtained as a colorless oil. Molecular weight 450.6 (mass spectrometry) Molecular formula C ₂₆ H ₄₂ O ₆ Infrared absorption spectrum (CHCl ₃ ): Figure 3 Nuclear magnetic resonance spectrum (CDCl ₃ ): Figure 4 Example 4 MB-530B carboxylic acid n-butyl ester MB- 200 mg of 530B was dissolved in 20 ml of n-butanol dehydrated with molecular sieve 3A, and a few drops of acetyl chloride were added thereto, followed by stirring at room temperature for 2 hours. The solvent was distilled off, and 50 ml of ethyl acetate was added for extraction.
The extract was washed with 2% aqueous sodium bicarbonate and saturated brine, dehydrated with Glauber's salt, and the solvent was distilled off. Silica gel prepared by drying with benzene (Wako Gel C-
The fractions eluted with a developing solvent of ethyl acetate-benzene (4:96) were collected, and the solvent was distilled off to obtain 80 mg of MB-530B carboxylic acid n-butyl ester as a colorless oil. . Molecular weight 478.7 (mass spectrometry) Molecular formula C ₂₈ H ₄₆ O ₆ Infrared absorption spectrum (CHCl ₃ ): Figure 5 Nuclear magnetic resonance spectrum (CDCl ₃ ): Figure 6 Example 5 MB-530B carboxylic acid benzyl ester MB-530B 200mg The mixture was dissolved in 20 ml of benzyl alcohol dehydrated with Molecular Sieve 3A, and several drops of acetyl chloride were added thereto, followed by stirring at room temperature for 3 hours. The solvent was distilled off, and 50 ml of ethyl acetate was added for extraction. The extract was washed with 2% aqueous sodium bicarbonate and saturated brine, dried over Glauber's salt, and the solvent was distilled off. Silica gel prepared by drying with benzene (Wako Gel C-
The fractions eluted with a developing solvent of ethyl acetate-benzene (4:96) were collected, and the solvent was distilled off to obtain 70 mg of MB-530B carboxylic acid benzyl ester as a colorless oil. Molecular weight 498.6 (mass spectrometry) Molecular formula C ₃₀ H ₄₂ O ₆ Infrared absorption spectrum (CHCl ₃ ): Figure 7 Nuclear magnetic resonance spectrum (CDCl ₃ ): Figure 8 Next, the following procedure was carried out according to the method of Example 2. Ta
An example of the method for producing MB-530B carboxylic acid ester is shown below. Note that the manufacturing method of MB-530B is the same as in Example 1. Example 6 MB-530B carboxylic acid methyl ester Crystal 500 obtained by culturing, extracting and concentrating in exactly the same manner as in Example 1, subjecting to high performance liquid chromatography, and repeating recrystallization with ethanol.
mg was dissolved in 50 ml of methylene chloride, about 20 ml of a saturated aqueous calcium hydroxide solution passed through a Millipore filter was added thereto, and the mixture was vigorously stirred at room temperature. When the pH of the solution drops to 8 or less, add the above calcium hydroxide aqueous solution 10ml at a time and continue stirring. When the pH no longer drops (total of 50ml of calcium hydroxide solution added), add distilled water and separate. The mixture was transferred to a liquid funnel, the solvent layer was collected, and the solvent was distilled off. N-heptane was added to the dried product, and the powder was ultrasonically treated, filtered and dried to obtain 450 mg of powder of MB-530B carboxylic acid calcium salt. The physical properties of the MB-530B carboxylic acid calcium salt thus obtained are as follows. 1 Molecular weight 441 (mass spectrometry) 2 Molecular formula C ₂₄ H ₃₇ O ₆ 1/2Ca 3 Melting point 155-165℃ (decomposition) 4 Specific rotation [α] _D = +209° (C = 1.48) 5 Infrared absorption spectrum ν _nax cm ^-1 : Measured using KBr pellets. 3400, 1730, 1580, 1440 6 NMR spectrum (CD ₃ OD, δ: ppm) 0.85
(6H, multiplet), 1.1 (6H, multiplet), 3.7 (1H,
multiplet), 4.2 (1H, multiplet), 5.35 (1H, Broadau multiplet), 5.5 (1H, Broadau multiplet), 5.9
(2H, multiplet) 500 mg of the MB-530B carboxylic acid calcium salt obtained in this way was dissolved in 5 ml of dimethyl sulfoxide, 0.3 ml of methyl iodide was added, and while stirring with a reflux condenser, the mixture was heated for 40 min. 5 at ~50℃
Allowed time to react. 5 ml of water was added to this reaction solution, extracted with 50 ml of dichloromethane, and the solvent was distilled off. The dried product was adsorbed on a silica gel column (Wako Gel C-100, 10 g) prepared with benzene, and ethyl acetate-
Fractions eluted with a developing solvent of benzene (4:96) were collected, and the solvent was distilled off to obtain 100 mg of oily MB-530B carboxylic acid methyl ester. The physical properties of the obtained MB-530B carboxylic acid methyl ester were the same as in Example 1. Next, the cholesterol synthesis inhibitory effect, serum cholesterol lowering effect, toxicity, effective dose, etc. of MB-530B carboxylic acid ester will be shown. 1 Cholesterol synthesis inhibitory effect MB-530B carboxylic acid ester is a rate-limiting enzyme in the cholesterol synthesis pathway, 3-hydroxy-3-methylglutaryl coenzyme A reductase [3-hydroxy-3-me-thylglutaryl]
(HMG)-Co A reductase]. Inhibits the enzyme activity by 50% as measured by the method described in the literature [Analytical Biochemistry (Anal.Biochem.) Vol. 31, p. 383 (1969)]
Table 1 shows the concentration of MB-530B carboxylic acid ester. Cholesterol synthesis inhibitory effect of MB-530B carboxylic acid ester [Journal of Biological Chemistry (J.Biol.Chem.) 247
Vol., p. 4914 (1972)] is extremely powerful. In other words, as shown in Table 1, the concentration that inhibits cholesterol synthesis by 50% is the same as that of the known strong inhibitor ML-236B (Japanese Patent Application Laid-open No. 155690-1982).
is about 10 ng/ml, while the methyl, ethyl and benzyl esters of MB-530B carboxylic acid have about 1 ng/ml, which is about 10 times more potent.

【table】

[Table] 2 Serum cholesterol lowering effect Triton WR1339 (trade name) (this substance has the effect of increasing blood cholesterol concentration) 400 mg/Kg to 5 rats per group (body weight approximately 300 g)
At the same time, MB-530B carboxylic acid ester was administered orally, and 20 hours later, the blood was exsanguinated to death.
The liver was collected and cholesterol was measured using a conventional method. The results are shown in Table 2.

[Table] Reduction rate 3 Acute toxicity An acute toxicity test was conducted by oral and intraperitoneal administration of MB-530B carboxylic acid ester (methyl, ethyl, n-butyl, benzyl) to mice. As a result, animals treated with these substances
The drug dose that causes 50% lethality (LD50) is 2000 mg/Kg or more for oral administration and 2000 mg/Kg or more for intraperitoneal administration.
At doses of 500 mg/Kg or more, the toxicity was extremely low. As is clear from the above test examples, MB-530B
Carboxylic acid esters have the effect of lowering blood lipids by inhibiting cholesterol biosynthesis, and can be used in medicine, for example, as antilipemic agents and arteriosclerosis preventive agents. MB-530B carboxylic acid ester can be administered orally or parenterally in the form of capsules, tablets, injections, and the like. Oral preparations are usually preferred. When used for treatment of adults, the dosage varies depending on the route of administration and the number of administrations, but
A range of 0.5 to 100 mg is preferred, such as 0.5 to 10 mg.

[Brief explanation of the drawing]

Figure 1 shows the infrared absorption spectrum of MB-530B carboxylic acid methyl ester, and Figure 2 shows the nuclear magnetic resonance spectrum of the same material. Figure 3 is MB
The infrared absorption spectrum of -530B carboxylic acid ethyl ester is shown, and Figure 4 shows the nuclear magnetic resonance spectrum of the same substance. Figure 5 shows the infrared absorption spectrum of MB-530B carboxylic acid n-butyl ester, and Figure 6 shows the nuclear magnetic resonance spectrum of the same material. Figure 7 shows the infrared absorption spectrum of MB-530B carboxylic acid benzyl ester;
The figure shows the nuclear magnetic resonance spectrum of the same material.

Claims (1)

[Claims] 1. MB-530B produced by culturing MB-530B-producing bacteria belonging to the genus Monascus and collecting MB-530B.
Characterized by reacting 530B with alcohol, the formula (In the formula, R represents an alkyl group: or a benzyl group that is unsubstituted or substituted with an alkyl group, an alkoxy group, or a halogen atom.)
MB-530B carboxylic acid ester manufacturing method. 2 The MB-530B producing bacterium belonging to the genus Monascus is Monascus anka.
SANK10171 (IFO6540), Monascus paxii SANK11172 (IFO8201), Monascus purpurous
SANK10271 (IFO4513), Monascus ruber SANK11272 (IFO9203), same
SANK17075 (CBS832.70), SANK17175
(CBS503.70), SANK17275 (ATCC18199), same
SANK15177 (Feikoken Jokyo No. 1340),
SANK13778 (Feikokenjoyori No. 1343),
SANK10671 (Feikokenjoyori No. 1342), same
SANK18174 (Feikoken Article No. 1341), Monascus vitreus
The manufacturing method according to claim 1, which is SANK10960 (National Health Examination 609, e-609) (Feikoken Jokyo No. 1344). 3 MB-530B-producing bacteria belonging to the genus Monascus were cultured, MB-530B was collected, and the obtained MB-
530B with a compound forming a metal salt, and then reacting the obtained MB-530B carboxylic acid metal salt with an alkyl halide or a benzyl halide, (In the formula, R represents an alkyl group: or a benzyl group that is unsubstituted or substituted with an alkyl group, an alkoxy group, or a halogen atom.)
MB-530B carboxylic acid ester manufacturing method. 4 The MB-530B carboxylic acid producing bacterium belonging to the genus Monascus is Monascus
anka) SANK10171 (IFO6540), Monascus paxii (Monascus paxii) SANK11172
(IFO8201), Monascus purpurous (Monascus purpurous) SANK10271
(IFO4513), Monascus Louver (Monascus
ruber) SANK11272 (IFO9203), SANK17075
(CBS832.70), same SANK17175 (CBS503.70), same
SANK17275 (ATCC18199), SANK15177 (Feikoken Co., Ltd. No. 1340), SANK13778 (Feikoken Co., Ltd. No. 1343), SANK10671 (Feikoken Co., Ltd. No. 1342)
(No.), SANK18174 (Feikoken Article No. 1341), Monascus vitreus (Monascus vitreus)
The manufacturing method according to claim 3, which is SANK10960 (National Health Examination 609, e-609) (Feikoken Jokyo No. 1344).