JPS59137443A - Metal salt and alkyl ester of dihydromonacolin l carboxylic acid and its preparation - Google Patents

Abstract

NEW MATERIAL:A metal salt or alkyl ester of dihydromonacolin L carboxylic acid of formula I [R is 1/n M (M is metal atom; n is valency of the metal atom) or lower alkyl]. EXAMPLE:Methyl dihydromonacolin L carboxylate. USE:It is used as a physiologically active substance having the action of lowering cholesterol. PREPARATION:A metal hydroxide of M(OH)n is made to act on dihydromonacolin L of formula II to give the titled compound of formula III. The compound of formula II is novel and obtained by collecting from the culture mixture of a strain in Monascus such as Monascus ruber No.1005 (FERM-P4882). The reaction is preferably carried out at 30-60 deg.C for several hours.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides novel dihydromonacolin L derivatives, more specifically, the following general formula (1), [wherein R 1/n
The present invention relates to metal salts and alkyl esters of dihydromonacolin L carboxylic acid represented by y (M represents a metal atom, n represents a valence of the metal atom) or a lower alkyl group, and a method for producing the same.

The present inventor previously discovered a bacterial strain belonging to the genus Monascus, such as Monascus rubel Nnl O05 [FeRM
-P Nn 4882)], we succeeded in isolating a new physiologically active substance having an excellent cholesterol-lowering effect, that is, dihydromonacolin L represented by the following formula (In, An application was filed (Japanese Patent Application No. 56-214788).

As a result of further research on this dihydromonacolin L, the present inventor discovered that the lactone ring of dihydromonacolin L was cleaved, and the metal salt of dihydromonacolin and carboxylic acid (1
, ) and alkyl ester (Ib) 75E, it was discovered that it has a cholesterol-lowering effect, and the present invention was completed.

(1,) (lb) (In the formula, R1 represents a lower alkyl group, and M and M are the same as above.) That is, the present invention provides metal salts and alkyl esters of dihydromonacolin L carboxylic acid having a cholesterol-lowering effect. This is what we provide.

Further, the present invention provides a method for producing metal salts and alkyl esters of dihydromonacolin carboxylic acid.

The metal salts of dihydromonacolin carboxylic acid of the present invention include alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, and aluminum salts, iron salts, zinc salts, copper salts, nickel salts and Examples include cobalt salts, among which alkali metal salts, alkaline earth metal salts and aluminum salts are preferred, and sodium salts, calcium salts and aluminum salts are most preferred.

Alkyl esters of dihydromonacolin L carboxylic acid include methyl ester, ethyl ester, H-
Examples include propyl ester, isopropyl ester, n-butyl ester, n-hexyl ester, and the like.

Metal salt of dihydromonacolin L carboxylic acid of the present invention (1
a) and the alkyl ester (Ib) (2) are produced by reacting dihydromonacolin L with a metal hydroxide of the formula (II'1) or an alcohol of the formula (IV), respectively.

M(OH)n (1> R, -0I((IV) (where Δ(, n and R1 are the same as above)) The raw material dihydromonacolin L was obtained by the method described in Japanese Patent Application No. 1988-214788. That is, a bacterial strain that produces dihydromonacolin L is aerobically produced in a nutrient medium by a method known as a mold culture method.

For example, dihydromonacolin L-producing bacteria produce soluble starch 2
%, glucose 1%, peptone 2%, and agar 2%, and dihydromonacolin is added to the culture medium.For production, the cells grown on this agar medium can be directly inoculated into a production medium and cultured. Furthermore, dihydromonacolin L can be produced there by culturing the m-body grown in a production medium in a new production medium.

Dihydromonacolin-producing bacteria grow at a temperature of 7 to 40°C, but a temperature of 20 to 35°C is usually preferred for production of dihydromonacolin L. To culture the Monascus bacteria that produces dihydromonacolin L, nutrients known for culturing molds and other microorganisms (available in many cases, such as glucose,
Maltose, dextrin, starch, lactose, zatucarose, glycerin, etc. can be used as carbon sources.

Among these carbon sources, glucose and starch are the preferred carbon sources for the production of dihydromonacolin.

Any known silicon source for the growth of Monascus or other microorganisms to produce dihydromonacolin L can be used. For example, peptone, meat extract, yeast, yeast extract,
Soybean flour, peanut flour, corn steep liquor, rice bran, nine inorganic sources, etc. can be used. When producing dihydromonacolin L by culturing dihydromonacolin-producing bacteria, inorganic salts and metal salts are added if necessary. Also, trace amounts of heavy metals can be added if necessary.

Dihydromonacolin L can be obtained by aerobically culturing its producing bacteria, and commonly used aerobic culture methods such as solid state culture, shaking culture, and aerated agitation culture may be 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 35°C. Culturing is continued for 4 to 15 days until substantial accumulation of dihydromonacolin L is achieved.

Dihydromonacolin L can be isolated from a culture by a combination of various methods depending on its properties. That is, for example, extraction with ether, ethyl acetate, chloroform, benzene, etc., dissolution in a highly polar solvent such as acetone or alcohol, removal of impurities with a less polar solvent such as petroleum ether or hexane, gel filtration with a Sephatex column, activated carbon, This includes adsorption chromatography using silica gel and the like. By using a suitable combination of these means, dihydromonacolin L can be isolated from the culture as a homogeneous substance.

To produce the metal salt (l&) of dihydromonacolin L carboxylic acid, dihydromonacolin L is dissolved in a solvent such as methanol, ethanol, acetone, benzene, etc., and an aqueous solution of metal hydroxide (1) is added thereto. The reaction may be carried out at ~60°C for several hours.

Alternatively, the alkyl ester (lb) of dihydromonacolin L carboxylic acid can be obtained by reacting cyclohydromonacolin L with the desired corresponding alcohol (IV). In this case, as a catalyst, an inorganic acid such as hydrochloric acid or sulfuric acid, or boron fluoride, or an acidic ion exchange resin is used, and as a solvent, one that does not participate in the reaction, such as the same alcohol, Matach-Hensen, chloroform, or ether, is used.

Dihydromonacolin L of the present invention thus obtained
The serum cholesterol-lowering effect and toxicity of metal salts and alkyl esters of carboxylic acids are as follows.

(11 Blood m cholesterol lowering effect Triton WR-1339 was intravenously injected at 400η/9 to rats, and at the same time, 9 was orally administered to 10ffiv/9 of dihydromonacolin derivative (dissolved in water or dissolved in water). 9 was orally administered to Low/, respectively.

Twenty hours after administration of Triton WR-1339, the mice were sacrificed by exsanguination and blood cholesterol was measured by a conventional method. The results are shown in Table 1.

Table 1. Serum cholesterol lowering effect on rats (2) Acute toxicity An aqueous solution or water suspension of the dihydromonacolin L derivative was administered orally or intraperitoneally to test for acute toxicity. The results are shown in Table 2.

Table 2 Dihydromonacolin L
The derivatives are administered by intravenous injection, oral transfusion, and the like. When used for the treatment of adults, the dosage varies depending on the administration route and the number of administrations, but is preferably in the range of 50 to 500 M9 per day, particularly 50 to 100 M9 per day.

The derivative of dihydromonacolin L, by analogy with other lipid-lowering agents such as clofibrate or synfiplate, can be made up to 4 g for administration in any conventional manner.

Therefore, the present invention also includes a pharmaceutical composition containing at least one derivative of dihydromonacolin L suitable as a human drug. Such compositions are provided for use in a conventional manner with any required pharmaceutical carriers or excipients.

Desirably, the composition is provided in a form suitable for absorption from the gastrointestinal tract. Tablets and capsules for oral administration are in unit dosage form and contain binders such as syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone, excipients such as lactose, sugar, corn starch, calcium phosphate, sorbitol, or It may contain conventional excipients such as glycine, lubricants such as magnesium stearate, talc, polyethylene glycol or silica, disintegrants such as potato starch, or acceptable wetting agents such as sodium lauryl sulfate. Tablets may be coated by methods well known in the art.

Oral liquid preparations may be aqueous or oily suspensions, solutions, syrups, elixirs, etc., or they may be dry products for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain commonly used excipients such as suspending agents such as sorbitol syrup, methyl cellulose, glucose/sugar syrups, gelatin, hydroxyebu cellulose, carboxymethyl cellulose, aluminum stearate gel or hydrogenated edible fats, emulsifiers, etc. lecithin, sorbitan monooleate or gum arabic, non-aqueous vehicles such as almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol, preservatives such as methyl p-hydroxybenzoate, propyl p-hydroxybenzoate or sorbic acid. May be contained.

Compositions for injection may be presented in focal dose ampoules or in multi-dose containers with an added preservative. The composition is a suspension,
They may take the form of solutions, emulsions in oily or aqueous vehicles and may contain formulation agents such as suspending agents, stabilizing agents, and (only) dispersing agents.

Alternatively, the active ingredient may be in powder form for reconstitution with a suitable vehicle, eg sterile pyrogen-free water, before use.

Next, the present invention will be explained in more detail with reference to Reference Examples, Examples, and Formulation Examples.

Reference Example 1 Glucose 3%, peptone 1%, defatted soybean flour 3%, glycerin 7%, NaN030.2%.

Monascus rubel Nn1O05 strain was inoculated into a liquid medium containing 1% MgSO4.7H200, and the temperature was 28°C.
The cells were cultured aerobically for 10 days. The obtained culture filtrate (50
t) was adjusted to pH 3 with 6N hydrochloric acid, and then extracted with an equal amount of ethyl acetate. The extract was concentrated to dryness and dissolved in 1 t of benzene to remove insoluble matter. The lachrymal fluid was washed twice with 500 ml of 5% sodium bicarbonate solution. Next, 1 part of 0.2N caustic soda solution was added to the benzene solution and stirred at room temperature.
After confirming by thin layer chromatography that dihydromonacolin L had disappeared from the benzene layer, the aqueous layer was collected. This aqueous layer was adjusted to pH 3 with 6N hydrochloric acid and extracted twice with 1 t of ethyl acetate. The extract was concentrated to dryness to obtain an oily substance 3.1.
I got 1. Monacolin K is precipitated at the same time as this oil is dissolved in benzene and crystallized. The mother liquor from which the crystals were removed was dried and 80F silica gel (Wakogel C-
200) was adsorbed onto the column.

The column was developed in the following order: 400 d of dichloromethane, 5 t of dichloromethane-ethyl acetate (9:1), and 7 t of dichloromethane-ethyl acetate (8:2). Dihydromonacolin L
402 silica gel (Wako Gel C) was dried.
-200) column and hexane 250rj!
, and 4 tons of hexane-acetone (9:1).

Both parts containing dihydromonacolin L were dried, dissolved in 60 tnl of benzene, and diluted with 5% bicarbonate of soda.
Washed twice.

Then, it was dehydrated and concentrated to dryness to obtain a white powder of 3011v. This white powder was further subjected to high performance liquid chromatography (silica gel-0DS system power +5'X).

After collecting the dihydromonacolin L fraction, the solvent was distilled off to obtain dihydromonacolin L 8 NV as a crystalline white powder.

Elemental analysis value theoretical value %: C74,50,H9,85 actual value %:
C74,63,H9,79 molecule fp306 (019H
3O03) Example 1 Dihydromonacolin L 12 oxqwobenzene 5ff
17! Dissolve in this and add 0. IN caustic soda aqueous solution 0.
4- was added and stirred at 40°C for 2 hours.

The aqueous layer was collected, washed three times with 2 Tnl of benzene, and then lyophilized to obtain 130 ml of white powder of sodium salt of dihydromonacolin L carboxylic acid.

Elemental analysis value theoretical value %: C65,87,H9,02 actual value %:
C66,16,H9,14 ultraviolet absorption spectrum (K
2O): Terminal absorption infrared absorption spectrum (KBr)
: 3420+2930.1720.16506n-”
Moreover, in the above embodiment, Ca(O
By using H)2 or Mg(OHh, calcium and magnesium salts were obtained, respectively.

Dihydromonacolin L90■ was dissolved in 10m of methanol, and an acidic ion exchange resin (Tauex 50, H
2001 ng of mold) was added and stirred at 60 to 70°C for 3 hours. After the reaction, the resin was removed and the solvent of the p solution was distilled off under reduced pressure.

The residue was purified by column chromatography (benzene-ethyl acetate = 9:1 and 8:2) using silica gel 10f to obtain dihydromonacolin L carboxylic acid methyl J ester 53q as an oil.

Elemental analysis value theoretical value %: c'o, c+7talO, 12 actual value %
: c71.37,1ilO,83 ultraviolet absorption spectrum (:EI20) : terminal absorption infrared absorption spectrum (KBr) : ~31001740.1725σ-
1 Formulation example 1 Oral capsule lactose 75
■ Magnesium stearate 15 ■ Total 340 mg After mixing the powder of the above formulation and passing it through a 60 mesh sieve,
340 kg of this powder was put into one gelatin capsule and made into a capsule.The applicant is Akira Endo.

Claims (1)

[Claims] l, the following general formula (1), [wherein R represents 1/n M (M represents a metal atom, and n represents the valence of the metal atom) or a lower alkyl group ] Metal salts and alkyl esters of dihydromonacolin L carboxylic acid represented by these. 2. React the metal hydroxide represented by the following formula (■) ('I) M(on)n (button (in the formula, M represents a metal atom and n represents the valence of the metal). A method for producing a metal salt of dihydromonacolin L carboxylic acid represented by the general formula (1m) (where M and n are the same as above). 3. A method for producing a metal salt of dihydromonacolin L carboxylic acid represented by the following formula (■) Formula () % Formula % () (In the formula, R1 represents a lower alkyl group)
) (wherein R1 is the same as above) A method for producing an alkyl ester of dihydromonacolin L carboxylic acid.