SE445828B - SALTS AND ESTERS OF MONACOLINE K FOR USE AS ANTI-HYPERCOLESTEROLEMIC AGENTS - Google Patents

Description

8065386-1 Preparation of salts and esters of monacolin K can take place by salt formation or esterification of monacoline K or a reaction-prone derivative thereof.

Furthermore, the production of monacolin K can take place by culturing a microorganism of the genus Monascus, which can produce monacolin K, and transferring monacoline K to a salt during or after the separation from the culture medium.

The salts of monacoline K contemplated by the present invention are metal salts and preferably alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts of Group IIIa metals of the Periodic Table such as the aluminum salt, and salts of transition metals from groups Ib, IIb and VIII of the Periodic Table, such as iron, nickel, cobalt, copper and zinc salts. Of these salts, alkali metal salts, alkaline earth metal salts and the aluminum salt are preferred, and the sodium, potassium and aluminum salts are most suitable.

Salts of monacoline K can be easily converted to monacoline K or to the parent acid of monacoline K by acidification. The resulting monacoline K or acid turns into a salt in the presence of alkali, e.g. an alkali metal hydroxide or carbonate.

This conversion can be performed quantitatively and repeatedly. It has been found that conversion between monacoline K or its parent acid and the metal salt is closely related to the pH of the medium containing them. The critical pH value is around 5.0. If metal ions are present, monacoline K is always present in the form of a salt, when the pH is above 5.0.

On the other hand, if the pH is less than 5.0, monacoline K or the parent acid of monacoline K or a mixture of both is present in varying conditions. 8005386-1 Estrons of monacoline K, prepared according to the present invention, are those compounds of the above formula, wherein R represents an unsubstituted alkyl group. When R represents an unsubstituted alkyl group, they may have straight or branched chain and suitably up to 8 carbon atoms. Examples of such alkyl groups R are methyl, ethyl, propyl, isopropyl, butyl and hexyl groups.

R may also represent a benzyl group.

Of the esters, the methyl, ethyl, butyl and benzyl esters are particularly preferred.

Salts and esters of monacoline K of the present invention can be readily prepared by salt formation or esterification of monacoline K or of a reaction-prone derivative of monacoline K.

Examples of suitable reaction-prone derivatives are the parent acid of monacolin K and, in the case of the preparation of esters, salts of monacoline K (eg the salt classes described above, in particular sodium, potassium, calcium, magnesium, aluminum, jane, 211m, Koppaw, nickel or koboic salts).

Salts of monacoline K can be prepared by simple reaction between monacoline K or its parent acid and an oxide, hydroxide, a carbonate or bicarbonate, preferably a hydroxide or a carbonate, of the selected metal. Care must be taken that the reaction is carried out at a pH in excess of 5.0, preferably in excess of 7.0, to obtain complete conversion of monacolin K or the acid to the desired salt. In this reaction, monacoline K used is conveniently prepared in a manner which are mentioned in the aforementioned patent applications, namely by culturing a fungus of the genus Monascus and isolating monacolin K from the culture medium, Monakolin K may be subjected to isolation from the culture medium and purified before salt formation or, preferably, the salt formation reaction is carried out during the isolation process. and the purification of monacoline K from the culture medium so that monacoline K is isolated in the form of the desired metal salt.Whatever method is used, the metal salt can be isolated from the reaction medium or the culture medium by methods well known in the art.

Esters of monacoline K can be prepared by simple esterification of monacoline K or a reaction-prone derivative thereof. The reaction can be carried out by reacting an alcohol of the formula ROH (or a reaction-prone derivative thereof), wherein R represents an alkyl group or benzyl group, with monacoline K or its parent acid in the presence of a hydrating agent, t. ex. an acid halide, ZS 40 8005386-1 such as nictile chloride. Alternatively, monacoline K esters can be prepared by reacting a salt of monacoline K with a halide of the formula RX (wherein R represents an alkyl group or benzyl group and X represents a halogen atom, preferably an iodine atom).

It is also possible to produce monacolin K directly by culturing a monacolin K-producing microorganism of the genus Monascus. Suitable microorganisms that can be used in this process are Monascus duck SANK 10171 (IFO 6540), Monascus purpurous SANK 10271 (IFO 4513), Monascus ruber SANK 10671 (Ferm 4958), Monascus vitreus SANK 10960 (NIHS 609, e-609 , Ferm 4960), Monascus paxii SANK 11172 (IFO 8201), Monascus ruber SANK 11272 (IFO 9203), Monascus ruber SANK 13778 (Ferm 4959), Monascus ruber SANK 15177 (Ferm 4956), Monascus ruber SANK 17075 (CBS 832.70) Monascus ruber SANK 17175 (CBS 503.70), Monascus ruber SANK 17275 (ATCC 18199) and Monascus ruber SANK 18174 (Ferm 4957). All these microorganisms are available from recognized cultural collections, which are listed below: IFO = Institute for Fermentation, Osaka, Japan Ferm = Fermentation Research Institute, Agency of Industrial Science and Technology, Ministry of International Trade and Industry, Japan NIHS = National Institute of Hygienic Sciences, Japan CBS = Centraal Bureau voor Schimmelcultures, The Netherlands ATCC = American Type Culture Collection, Maryland, USA Suitable strains of the genus Monascus, which can be used according to the present invention for the production of monocholine K, are Monascus ruber SANK 10671, Monascus ruber SANK 11272, Monascus ruber SANK 13778, Monascus ruber SANK 15177 and Monascus ruber SANK 18174. monascus ruber SANK 10671, Monascus ruber 13778, Monascus ruber SANK 15177 and Monascus ruber SANK 18174 are all fungi, recently isolated from soil by inventors to the present invention and the microbiological properties are set forth below: Monascus ruber SANK 15177 (Ferm 4956) This strain was isolated from soil in Tukimíno, Yamato-city, Kanagawa Prefecture, Japan, and deposited on 27 April 1979 under accession number 4956 with the Fermentation Research Institute.

The growth of the strain is good on potato-glucose-agar medium at -25 ° C and produces a soluble, coloring material-with a yellowish-brown to reddish-brown color in the medium. Many kleistotecies are formed on the hyphae layer.

On oatmeal agar medium a pale, brown coloring material is formed and the growth is good. The formation of kleistothecia is good and the kleistothecia are spherical with a diameter of 30-60 μm and. formed on short stems. These stems are almost colorless and branched with a size of 25-60 x 3.5-5.0 μm. Asci soon disappear and are difficult to observe. The ascospores are colorless and ellipsoidal and their dimensions are 4.5-5.5 x 4.0-5.0 μm, the surfaces are smooth. The conidia are bound base epitally and are of the size 7.0-10.0 X 6.0-10.0 μm. Their tissues are ruptured.

Although the stem grows at 37 ° C, the best growth is between 23 and SOOC.

Monascus Ruber SANK 10671 (Ferm 4958) This strain was isolated from soil in Shinagawa-ku, Tokyo, Japan, and deposited on April 27, 1979 with the Fermentation Research Institute under Accession No. 4958.

Growth on potato-glucose-agar and oatmeal-agar media is similar to that of strain SANK 15177 except that the soluble, coloring material is dark red. The diameter of the kleistotecier is 30-80 pm and the dimensions of the stems are 30-70 X 3.0-5.0 pm.

Asci is not observed. The ascospores are colorless and ellipsoidal and their dimensions are 4.5-6.5 X 4.0-5.0 μm. The conidia are colorless and pear-like or egg-like and their dimensions are 6.0-10.0x 6.0 ~ 8, S um.

Monascus Ruber SANK 13778 (Ferm 4959] This strain was isolated from soil in Inawashiro-cho, Nagata, Yama-gun, Fukushima Prefecture, Japan, and deposited on 27 April 1979 with the Fermentation Research Institute under Accession No. 4959.

Growth on potato-glucose-agar and oatmeal-agar media is similar to that of strain SANK 15177 with the exception of the soluble reddish-brown to reddish-brown color.

The kleistotecies have a diameter of 35-75 μm and the large, coloring material of the stems has a play is 30-70 x 3.5-5.0 μm. Asci is not observed. The ascospores are colorless and ellipsoidal with the dimensions 4.5-6.0 x 4.0-5.0 μm.

The surfaces are smooth. The dimensions of the conidicides are 7.0-10.0 x 6.0-10.0 nm. 8035386-1 6 Monascus ruber SANK 18174 (Ferm 4957) This strain was isolated from soil in Shakotan-cho, Shakotangun, Shiribcshí Shicho, Hokkaido Prefecture, Japan, and deposited on April 27, 1979 at the Fermentation Research Institute under accession number 4957.

Growth on potato-glucose-agar and oatmeal-agar media is similar to that of strain SANK 15177 except that the dyeing material produced is pale red The Kleistotecia have a diameter of 20-70 μm and the dimensions of the stems are 20-60 X 3 , Uß, 0 Um. Fl ßciobserveras ej. The ascospores are colorless and ellipsoidal with the dimensions 5.0-7.0 X 4.0-5.5 μm and their surfaces are smooth.

The conidia are bonded basipetally and are colorless and most of them are pear-like with the dimensions 6.0-9.5 X 6.0- .0 μm.

Based on the previous observations of the characteristics of the microorganisms, all were identified as strains of Monascus ruber van Tieghem.

Microbiological properties of Monascus ruber have been described in the following literature: Takada, Transactions of the Mimlogical Society of Japan, 2, 125-130 (1969) [Materials for the Fungus Flora of Japan (7)] and van Tieghem, Bull.Soc.Botan .France, eel, 227 (1884). Ascosporin formation of the strain has been described by Cole et al in The Canadian Journal of Botany, ig, 987 (1968), "Conidium 'ontogeny in hyphomycetes. The imperfect state of Monascus ruber and its meristem arthrospores".

In addition to the mems listed above, other fungi of the genus Monascus incl. Variants and mutants that can produce Monakolin K are used according to the present procedure.

Monacolin K can be prepared by culturing the selected microorganism in a nutrient medium under aerobic conditions using the same technique well known for culturing fungi and other microorganisms. The selected strain of Monascus can first be grown in a suitable medium, after which the produced microorganisms can be collected and inoculated into and grown on another culture medium for the production of monacolin K. The nutrient medium used to propagate the microorganism and the The nutrient medium used for the production of monacolin k 'may be the same or different. Each within 8085386- 1 7. This well-known culture medium for growing fungi can be used provided that it contains in a known manner the required nutrient materials, in particular an assimilable carbon source and an assimilated nitrogen source. Examples of suitable sources of assimilable carbon are glucose, maltose, dextrin, starch, lactose, sucrose and glycerol. Of these sources, glucose and glycerol are particularly suitable for the production of monacolin K.

Examples of suitable sources of assimilable nitrogen are peptone, meat extract, yeast, yeast extract, soybean meal, peanut meal, corn steep liquor, rice bran and inorganic nitrogen sources. Of these nitrogen sources, peptone is particularly suitable. When monacoline K is produced, an inorganic salt and / or a metal salt may be added to the culture medium. Furthermore, a smaller amount of a heavy metal may also be added. When producing salts of monacolin K by fermentation with a fungus of the genus Monascus, it is important that metal ions are present in the culture medium or inside the fungal body, which correspond to the metal salt to be produced.

It is particularly convenient that the microorganism is first grown on a potato dextrose agar medium (eg a product from Difco Company) and then inoculated into and grown on another culture medium to produce the desired salt of monacoline K.

The microorganism is suitably grown under aerobic conditions using per se known culture methods, e.g. cultivation with solid substrates, cultivation under shaking or aeration and stirring. The growth of the microorganism takes place over a wide temperature range, eg-7-35 ° C, but especially in the production of monacolin K, the most suitable culture temperature is 20-30 ° C.

During the culture of the microorganism, the production of monacoline K can be monitored by sampling the culture medium and measuring the physiological activity of monacoline K in the culture medium by tests described below. The culture can then be continued until a considerable accumulation of monacoline K is reached in the culture medium. At this time, monacoline-K can then be isolated and recovered from the culture medium by any suitable combination of isolation techniques, which are selected taking into account physical and chemical properties.

LH 8005386-1 a Any or all of the following isolation methods may be used, for example: extraction of the liquid from the nutrient medium with a hydrophilic solvent (eg diethyl ether, ethyl acetate or chloroform); extraction of the organism with a hydrophilic solvent (such as acetone or an alcohol); concentration, e.g. by evaporating all or part of the solvent under reduced pressure; solution in a more polar solvent (such as acetone or an alcohol); removal of impurities with a less polar solvent (such as petroleum ether or hexane); gel filtration through a column of a material such as Sephadex (trademark for a material marketed by Pharmacia Co. Limited, USA); absorbent chromatography with activated carbon or silica gel; rapid liquid chromatography; conversion to monacolin K or its parent acid; direct purification in the form of the metal salt and other similar methods. By using a suitable combination of these methods, monacoline K can be isolated from culture fluid. can as a pure substance.

The physiological activity of monacholine K salts and esters can be analyzed and determined quantitatively by the following tests, which can also be used in modified form to monitor the production of monacoline K salts during the fermentation process of the present invention. 1. Inhibition of Cholesterol Biosynthesis Like monacoline K, salts and esters of monacolin K can specifically inhibit the activity of 3-hydroxy-3-methylglutaryl-CoA reductase, which is the formation-determining enzyme in cholesterol biosynthesis.

The following Table 1 lists the concentrations (in ng / mg) of the compounds of the invention which inhibit the activity of 3-hydroxy-3-methylglutaryl (HMG) -CoA reductase by 50% [measured according to the method described in Analytical Biochemistry, eel, 383 (1969) | and the concentrations (in ng / mg) of the compounds of the invention which inhibit cholesterol biosynthesis with SO% fi according to the method described in Journal of Biological Chemistry, gíl, 4914 (1972)]. Furthermore, corresponding results are given for the known compound ML-256B (which is a compound which is known to have similar activity and which is prepared by culturing organisms of the genus Penicillium. It is described in British Patent Specification 1453425). The concentration of monacolin K that inhibits cholesterol biosynthesis by 50% is also indicated. It appears. that while the required concentration of NL-236B to inhibit cholesterol biosynthesis by 50% is 10.0 ng / ml, the corresponding concentration for monacoline K esters is of the order of 1 ng / ml (ie about 10 times better) and the corresponding concentration of the sodium salt of monacolin K is about 0.14 ng / ml (ie about 70 times better). The activity of the salts and esters of monacolin K is comparable to or better than that of monacolin K. Table 800 Compound I Required concentration (ng / ml) š ¶_ for inhibition with so s 1 É HMG-coA- I cholesterol - É; reductase biosynthesis 1 Methyl ester 70.0 1 1, z 1 Ethyl ester § 12.0 1 1.3 'Butyl ester 1 15.0; 2.0 Benzyl ester 1 11.0 1 1.1 Sodium salt 1 2.0 É 0.14 Calcium salt 12.0 ¿1.8 1 Mønacoiin K - É 2.0 ML-2363 10.0 10.0 2. Reduction of blood cholesterol levels of the Imamichí strain, each with a body weight of about 300 g.

The tests were performed on groups of rats with 5 animals in each group. Each animal was injected intravenously with 400 mg / kg of Triton WR-1339 (a trademark for a substance which increases blood cholesterol levels) and at the same time one of the compounds listed in the following Table 2 was orally administered in the amount indicated in the table. After oral administration, the rats were sacrificed by bleeding, and blood and liver were collected and cholesterol levels were determined in a conventional manner. The results are shown in Table 2, which shows the reduction in cholesterol levels in the blood and liver, compared with a control group of rats, which were only given Triton WR-1339.

The animals used in this test were rats of Wister 8005386-1. For comparison, similar results are given for Monakolin K and for ML ~ 236B, but these compounds were administered in considerably higher doses than those given by the compounds of the invention to achieve a comparable reduction. in cholesterol content.

Table 2 Compound Dose Reduction in cholesterol content in% mg / kg blood I liver Methyl ester s 23.8 μl 18.4 Ethyl ester 5 23.3 3 18.0 Butyl ester 5 19.5 o 15.7 Benzyl ester 5 24.4 2 18, 5 Sodium salt 2 27.5} 18.9 Cesium salt s 21.4 É 11.1 ~ 1 nonacoin K 10 _ 22.4 § 16.7 ML-zsaß 40 g 24.6 Å 20.9 The reduction of cholesterol in the blood or liver is obtained 1 - (C - B) A - B content in group treated only with Triton WR-1339 by the formula x 100, wherein content in untreated control group _ content in test group Acute toxicity The compounds tested were methyl, ethyl, butyl and benzyl esters and sodium and calcium salts of monacolin K. It was found that the 50% lethal dose (LD50) for each of these compounds was at least 2000 mg / kg when administered orally and at least 500 mg / kg when administered intraperitoneally. The compounds thus have a very low toxicity.

These results show that the compounds of the present invention inhibit cholesterol biosynthesis and thereby lower blood cholesterol levels. Thus, they are valuable drugs in the treatment of hyperlipemia and arteriosclerosis.

The compounds of the present invention may be administered orally or parenterally in the form of capsules, tablets, injectable preparations or other known preparations. Normally, however, oral administration is preferred. The dose varies depending on the age and body weight of the patient and the severity of the condition, but in general the daily dose for an adult is suitably 0.1-100 mg, preferably 0.1-10 mg, in the case of monacolin. K salts and 0.5-100 mg, preferably 0.5-10 mg, in the case of esters of monacolin K.

The preparation of the compounds according to the invention is further illustrated with reference to the following examples.

Example 1. The sodium salt of monacolin K 300 liters of a culture medium of pH 5.5 before sterilization containing 5% (w / v) glucose, 0.5% (w / v) corn steep liquor, 2% (w / v) peptone (Kyokuto brand, available from Kyokuto Seiyaku KK, Japan) and 0.5% (w / v) ammonium chloride were charged into a 600 liter fermentation vessel and inoculated with the organism Monascus ruber SANK 18174 (Ferm 4957).

Culture of the organism is carried out for 116 hours at 26 ° C with an aeration of 300 liters / min and stirring at 190 rpm. At the end of this time, the pH of the organism containing the culture broth was adjusted to 3.4 by the addition of 6N hydrochloric acid, after which the culture broth was extracted with 800 liters of methanol. 6 kg of hyflosupercel were added and then the extract was filtered using a filter press, whereby 1100 liters of methanolic extract were obtained. This extract was washed with 200 liters of saturated aqueous sodium chloride solution and then with 180 liters of ethylcyclohexane. The resulting solution was extracted with 600 liters of ethylene dichloride. 50 g of trifluoroacetic acid was added to the ethylene dichloride extract and left to react at 80 ° C for 30 minutes. The reaction mixture was then washed successively with 200 liters of a 2% (w / v) aqueous solution of sodium bicarbonate and with 200 liters of a 10% (w / v) aqueous solution of sodium chloride. The mixture was then concentrated by evaporation under reduced pressure to give 135 g of an oily substance.

This oily substance was dissolved in 400 ml of methanol. 20 ml of the resulting methanolic solution (containing 6.8 g of the oil) was then subjected to rapid preparative liquid chromatography using equipment equipped with a pre-coating of a Waters Co. Limited System 500 pac C18 and water (8S: 15 volume) were used as developer. Development was performed at a flow rate of 200 ml / min (development time about 10 minutes) while observing a connected differential recolumn (reversed phase column). A mixture of methanol fractometer and the fraction showing the major peak of the differential refractometer was separated. This operation was repeated Z0 800-5386-1 'W' in N 12 and the resulting major peak fractions were collected and concentrated to give 10.2 g of an oily substance. This oily substance was dissolved in 30 ml of methanol, and 6 ml of the methanolic solution (containing about 2 g of the oil) was subjected to the same preparative liquid chromatography and developed with a mixture of methanol and water (80:20 volume) at a flow rate of 200 ml / min. The fraction showing the main peak was separated. This operation was repeated and the main peak fractions were collected and concentrated. The residue was treated with aqueous methanol to give 1170 mg of crude crystallizer, which was recrystallized several times from ethanol to give 864 mg of crystals.

To these crystals was added 19.4 ml of 0.1N aqueous sodium hydroxide, and the mixture was stirred at 50-60 ° C for 3 hours. Insoluble material was filtered off and the filtrate was lyophilized to give 900 mg of the sodium salt of monacoline K having the following properties: 1. Color and shape White powder 2. Blementar analysis calculated C 64.84% H 8.38% Na 5.27% found C 64.78% H 8.55% Na 5.21% 3. Molecular weight 444 Through mass spectrometry (FD Mass.)] 4. Molecular formula C24H3706Na. Ultraviolet absorption spectrum: see Fig. 1 of the accompanying drawing, Amaxz 232 nm (log e = 4.30) 239 nm (log e = 4.36 248 nm (log e = 4.19) O \ __Infrared absorption spectrum (KBR- (tablet): see Fig. 2 of the accompanying drawing \ 1 .NMR spectrum (D20): see Fig. 3 of the attached drawing OO Preparative liquid chromatography: Residence time 8.5 min column, Microbondapac C 18 60% v / v aqueous with - tanol + 0.1% PIC-A (a product of Waters Co.Limited) Flow rate, 1.5 ml / min See Fig. 4_in attached drawing 9. Solubility: Soluble in water, insoluble in organic solvents Specific rotation: [α] š5 = + 266 ° (c = 0.33, water) I 8005386-1 Sodium salt of monacolin K 300 liters of a culture medium of pH 7,4 before sterilization, containing 1,5% (w / v) soluble starch, 1.5 5 w / v) glycerol, 2.0% (w / v) fishmeal and 0.2% (w / v) calcium carbonate were charged into a fermentation vessel of - 600 liters and inoculated with the organism Monascus ruber SANK 17075 (CBS 832701) Cultivation of organism one was performed for 120 hours at 26 ° C and aeration at 300 liters / min and stirring at 190 rpm. The culture medium was then filtered using a filter press to obtain 35 kg (wet weight) of the organism. 100 liters of water were added and the pH of the mixture was adjusted to 12 by adding sodium hydroxide with stirring. The mixture was then allowed to stand at room temperature for 1 hour, after which 2 kg of hyflosupercel was added to the mixture, which was then filtered in a filter press. The pH of the filtrate was adjusted to 10 by the addition of hydrochloric acid, and the resulting mixture was then adsorbed on a column containing 5 liters of HP-20 resin, which was washed with 15 liters each of water and 10% (v / v) aqueous methanol. . The column was then eluted with 90% (v / v) aqueous methanol. The eluate was concentrated to a volume of about 10 liters and then the pH was adjusted to 2 by adding hydrochloric acid and the mixture was extracted with ethyl acetate. The extract was washed with a saturated aqueous solution of sodium chloride, dried over anhydrous sodium sulfate and then concentrated by evaporation to dryness. to 50 g of an oily substance containing monacolin K acid. To this oily substance was added methanol to a volume of 100 ml. 20 ml of the resulting solution were subjected to preparative liquid chromatography using a reversed phase column (according to Example 1), eluted with a 20% (v / v) aqueous solution of methanol (containing 2% acetic acid) at a flow rate of 200 ml / min. The fraction showing the peak of the differential refractometer was separated (7-10 min). The remaining 80 ml of the methanolic solution was then treated in the same manner. The resulting major peak fractions were collected, concentrated and extracted with ethyl acetate. The extract was concentrated to dryness after addition of heptane to give 1.2 g of an oily substance. -acid was obtained. To this was added 2 ml of methanol and 100 ml of water, and the resulting solution was adjusted to pH 8.0 by adding 1N aqueous sodium hydroxide, so that a clear aqueous solution was obtained. This solution was passed through a column containing 10 ml of HP resin and the column washed was then washed with 80% (v / v) aqueous methanol.

The eluate was lyophilized to give 130 mg of the sodium salt of monacoline K as a white powder. The properties of the product were identical to those of the product of Example 1.

Emmpelö. Calcium salt of monocolin K Culture, extraction, concentration, rapid preparative liquid chromatography and recrystallization from ethanol were repeated according to Example 1. 500 mg of the resulting crystals were dissolved in 50 ml of methylene chloride and the resulting solution was filtered through a milliphoric filter. 20 ml of a saturated aqueous solution of calcium hydroxide was added to the filtrate, after which the mixture was stirred vigorously at room temperature. When the pH of the solution dropped below 8, an additional 10 ml of saturated aqueous calcium hydroxide solution was added and stirring was continued. When the pH no longer dropped (after adding a total amount of 50 ml of the aqueous solution of calcium hydroxide), distilled water was added and the whole solution was introduced into a separatory funnel. The organic phase was collected and the solvent was distilled off. The residue was further treated with heptane to give a powder by subjecting the mixture to ultrasonic treatment. The powder was filtered and dried to give 450 mg of the calcium salt of monacolin K as a white powder.

The calcium salt had the following properties: 1.hblekyl weight: 882 (according to mass spectrometry). hbalkyl formula: (C24H3706) 2fCa l \) 3. Melting point: 155-165 ° C (with decomposition 1,48, chloroform) ll 4. specific rotation: [ajšs = + zo9 ° (c. Infrared absorption spectrum (KBr): (see Fig. 5 in the accompanying drawing) '6. NMR Spectrum (CD30D): see Fig. 6 in the attached drawing Z0 8005386-1 Exemnel 4. Methyl ester of monacoline K Culture, extraction, concentration, fast gripping liquid chroma tography and recrystallization from ethanol were repeated according to Example 1 to give 864 mg of monacoline K. 200 mg of this monocoline K were dissolved in 20 ml of methanol (previously dehydrated with molecular sieve SA) After the addition of a few drops of acetyl chloride, the solution was stirred. at room temperature for 3 hours. The solvent was distilled off and the residue was extracted with 50 ml of ethyl acetate.

The extract was washed successively with a Z% sodium bicarbonate solution and a saturated aqueous solution of sodium chloride, and then dried over anhydrous sodium sulfate, after which the solvent was distilled off. The residue was adsorbed in a column containing 10 g of silica gel (Wakogel C-100), previously treated with benzene. The fractions were eluted with a mixture of ethyl acetate and benzene (6:94 v / v) was collected and the solvent was distilled off to give 65 mg of the methyl ester of monacoline K as a colorless oil.

This product had the following properties: 1. Molecular weight 436.5 (according to mass spectrometry) 2. Molecular formula: C25H4006 3. Specific rotation: [a] å5 = + 208 ° (c = 1.05, methanol) 4. Infrared absorption spectrum: see Fig. 7 in the accompanying drawing. NMR spectrum: see Fig. 8 of the accompanying drawing Example 5. Methyl ester of monacoline K The sodium salt of monacoline K was prepared 100 mg of this sodium salt was then dissolved in 2 ml of dimethyl sulfoxide, and then 50 μl of methyl iodide was added to the solution. The solution was heated under reflux with stirring at 40 DEG-50 DEG C. in a reactor equipped with a reflux condenser. The reaction mixture was then diluted with 5 ml of water and extracted with 10 ml of methylene chloride. The solvent was distilled off from the extract and the residue was adsorbed in a column containing 10 g of silica gel (Vogogel C-100), previously treated with benzene. The fractions eluted with a mixture of ethyl acetate and benzene (4:96 v / v) were collected and the solvent was distilled off to give mg of the methyl ester of monacoline K as an oil. according to Example 2. 8005386-1 Example 6. Ethyl ester of monacoline K 100 mg of monacoline K (prepared according to Example 1] was dissolved in ml of ethanol, previously dehydrated with molecular sieve 3A, and a few drops of acetyl chloride were added to the solution. h at room temperature, the solvent was distilled off and the residue was extracted with 50 ml of ethyl acetate, the extract was washed successively with a 2% (w / v) aqueous solution of sodium bicarbonate and a saturated aqueous solution of sodium chloride and then dried over anhydrous sodium sulfate. .

After distilling off the solvent, the residue was adsorbed in a column containing 10 g of silica gel (Wakogel C-100), previously treated with benzene. The fractions were eluted with a mixture of ethyl acetate and benzene (6:94 v / v) were collected and the solvent was distilled off to give 30 mg of the ethyl ester of monacoline K as a colorless oil.

This compound has the following properties: 1. Molecular weight: 450.6 (according to mass spectrometry) 2. Molecular formula C26H4Z06 3. Infrared absorption spectrum (CHCl3); see Fig. 9 of the attached 1 drawing 4. NMR spectrum (CDCl 3); see Fig. 10 of the accompanying drawing Example 7. Butyl ester of monocolin K 200 mg of monacoline K (prepared according to Example 1) was dissolved in ml of butanol, previously dehydrated with molecular sieve SA. After adding a few drops of acetyl chloride to the solution, the mixture was stirred at room temperature for 2 hours. The solvent was then distilled off and the residue was extracted with 50 ml of ethyl acetate. The extract was washed successively with a 2% (w / v) solution of sodium bicarbonate and a saturated aqueous solution of sodium chloride. The solution was then dried over anhydrous sodium sulfate and the solvent was distilled off. The residue was absorbed in a column containing 10 g of silica gel (Wakogel C-100), previously treated with benzene. The fractions were eluted with a mixture of ethyl acetate and benzene (4196 v / v) and collected, after which the solvent was distilled off. 80 mg of the butyl ester of Morakolin h were obtained in the form of a colorless oil. 8005386-1 17 The compound had the following properties: 1. Molecular weight 478.7 (according to mass spectrometry) 2. Molecular formula: C28H46O6 3. Infrared absorption spectrum (CHCl3): see Fig. 11 in the attached drawing * '4. NMR spectrum (CDC13): see Fig. 12 in the accompanying drawing.

Example gel 8. Benzyl ester of monacolin K 200 mg of monacolin K (prepared according to Example 1) were dissolved in 20 ml of benzyl alcohol, which had previously been dehydrated with molecular sieve SA.

After adding a few drops of acetyl chloride to the solution, the resulting mixture was stirred for 3 hours at room temperature.

The solvent was then distilled off and the residue was extracted with 50 ml of ethyl acetate. The extract was washed successively with a 2% (w / v) aqueous solution of sodium bicarbonate and a saturated solution of anhydrous sodium sulfate, and the solvent was distilled off from aqueous sodium chloride solution. The solution was then dried. The residue was adsorbed in a column containing 10 g of silica gel (Wakogel C-100), previously treated with benzene. The fractions eluted with a mixture of ethyl acetate and benzene (4296 v / v) were collected and the solvent was distilled off to give 70 mg of the benzyl ester of monacoline K as a colorless oil.

The compound had the following properties: 1. Molecular weight 512.6 (by mass spectrometry) Z. Molecular formula C31H44O6 3. Infrared absorption spectrum (CHCl): see Fig. 13 in the attached 3 drawing J>. NMR spectrum (CDCl 3): see Fig. 14 of the accompanying drawing J

Claims (4)

8005386-1 Patent claims 1. I. Compounds for use as non-hypocholesterolemic agents, characterized in that the H0 coo (R) n on \\\ l \ wherein R represents an alkali metal, an alkaline earth metal, a metal from group IIIa, Ib, IIb or in the periodic table of the elements, a C1-8 alkyl group or a benzyl group; and learns a number corresponding to the valence of the atom or group- has the formula: // fi \ T / ”\\ pen, which corresponds to R. Compounds according to claim 1, characterized in that R represents sodium, potassium, calcium, aluminum, iron, zinc, copper, nickel or cobalt. Compounds according to claim 1, characterized in that R represents a methyl, ethyl, propyl, isopropyl, butyl or hexyl group. Compounds according to claim 1, characterized in that R represents a benzyl group.