JPS5890509A - Composition for remedy of hypercholesterolemia - Google Patents

Abstract

PURPOSE:To provide the titled composition composed of a specific agent for inhibiting the synthesis of cholesetrol and a non-toxic basic anion exchange resin which bonds with cholic acid in the gastroenteric system and makes the acid unabsorbable through the system. CONSTITUTION:A composition for remedy of hypercholesterolemia is prepared from (A) an agent for inhibiting the synthesis of cholesterol, selected from the carboxylic acid of formulaIor formula II (R<1> is H or methyl; R<2> is H or methyl; R<3> is OH or methoxy), its salt, its lower alkyl ester, or its lactone, and (B) a non-toxic basic anion exchange resin which bonds with cholic acid in the gastroenteric system and makes the acid unabsorbable through the system (e.g. a resin containing the structural group of formula III and having a size of 50- 100 mesh). The compound of formulaIor formula II can be prepared from the compound of formula IV or formula V by the microbial enzymatic hydroxylation. Dose: 10-100mg daily in 3-4 divided doses.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a) formula (wherein sR'Fi represents a hydrogen atom or a methyl group, and R2
represents a hydrogen atom or a methyl group, and R' represents a hydroxyl group or a methoxy group. and b) a pharmaceutically acceptable non-toxic basic ion exchange resin that is capable of binding bile acids in the gastrointestinal system and making reabsorption impossible. relating to things.

The present inventors have demonstrated that co-administration of a new specific cholesterol synthesis inhibitor represented by formula (1) or expansion 1) and a certain bile acid absorbent can significantly reduce blood cholesterol levels. found that it works.

The cholesterol synthesis inhibitor used in the present invention is a carnoic acid having the formula (13 or It has the effect of specifically inhibiting the action of a certain 3-hydroxy-3-methyldaltaryl-Com reductase.

Pharmacologically acceptable salts of cardanic acid having the above formula (1) or above, such as alkali metal salts such as sodium or potassium; lower alkyl esters of carnoic acid having the above formula (1) or below) , such as methyl, ethyl, butyl, isopropyl, butyl, isobutyl, etc.

The lacto/isomer of the compound having the formula (1) or (2) is a compound in which the substituent at the 8-position in hexahydronaphthalene has the following partial structural formula.

In the above formula (1), the compound in which R1 is a hydrogen atom is 3-hydroxy-ML-236Bli, and the compound in which the substituent at the 3rd position is -OH is M-4g, -...O
The compounds of R are abbreviated as M-4's. This compound is disclosed in Japanese Patent Application No. 55-124385 and Japanese Patent Application No. 56-10596.
It is described in the specification of No. 7 along with its manufacturing method.

In the formula (1) above, the compound in which R1 is a methyl group is hereinafter abbreviated as 3-hydroxy-MB-530B@.

This compound is described in Japanese Patent Application No. Sho 56-134558, together with its manufacturing method.

In the above formula 〇), the compound in which R2 is a hydrogen atom and R5 is a hydroxyl group is hereinafter referred to as 6-hydroxyin ML-
It is abbreviated as 23681i. This compound was patented in 1983.
15483 and Japanese Patent Application No. Sho-55-124385 together with its manufacturing method.

A compound in which B2 is a water atom and R3 is a methoxy group in the above formula ω is hereinafter referred to as 6-medoxyisoM
It is abbreviated as L-236BII. This compound was patented in 1983.
-114038 #4 IIIB together with its manufacturing method.

In the above formula, the compound in which R2 is a methyl group and B5 is a hydroxyl group is hereinafter referred to as 6-hydroxyinoMB-
It is abbreviated as 5308m. This compound was patented in 1986-13.
It is described in the specification of No. 4558 along with its manufacturing method.

The compound having the above formula (13 and 013 is M of the following formula)
It is obtained by enzymatic hydroxylation using microorganisms using L-2368 or MB-530B or derivatives thereof as a raw material compound according to the method described in the above patent application.

ML-2368MB-530B Production examples of these compounds will be described below.

Structure example IM-4' type (1) 500- containing medium 100- of the following composition
Synthephalastrum nigricans 5ANK 42372 was inoculated into 20 E-square flasks and incubated at 26°C and 220°C.
r+p, ff1. After 3 days, ML-23
68 lactone was added at a final concentration of 0.05, and the mixture was further cultured at 26° C. and 220 r, p, m for 6 days.

Medium composition Glucose 1.0 tons - 4 f tons 0.2 Meat extract
o, i Yeast extract 0.1 Corn stag liquor 0.3 Tap water
Remaining (-uncorrected) After completion of the culture, the conversion reaction solution was treated with P, and F* was adjusted to PH3 with triple oroacetic acid. It was then extracted three times with %lj of ethyl acetate to obtain a fraction containing M-4'-hkdf.

Physical properties of M-4'cal&:/acid 1) TLC TLC plate: Merck Silkal Art5715 Solvent: Benzene; Acetone: Vinegar al=50:50:3R
1 value 0.46 (2) The extract of (1) was washed with saturated saline, and (i
After dehydration with lIm2 sodium, catalytic amount of trifluoroacetic acid was added for tectonication. Next, the above extract was diluted with 5
It was washed with an aqueous sodium bicarbonate solution, dehydrated with mal sodium, and then dried under reduced pressure. Next, the obtained residue was eluted with benzene:acetone=7:3 using a Roper column (manufactured by Merck & Co., Ltd., 816G Nize A) to obtain M-4'.
The lactone body was collected and further recrystallized using ethyl acetate to obtain about 180q of M-4' lactone body.

Physical properties of M-4' lactone 1) NMI spectrum In deuterated chloroform, using τMS as an internal standard, 10
Measured at 01als. (cocts, δ: ppm)
4.25 (IH, multiple wire) 4.60 (l) l, multiple wire) 5.5 (j (IH, multiple 111i 5.75 (l) i/multiple wire) 5.90 (IH, four wire) Iwao) 6.01 (IH, dual 2) ultraviolet absorption spectrum (methanol solution) λwax
(am): 230 z 237 + 2453) Infrared absorption spectrum (KBr method) cIIL-1:350
0.1720 4) Mass spectrum rIA: 406CM"), 304; 286
5) Optical rotation [α) D: +310.9° (C=0.66, / I
/-ru) 6) Melting point 141-143℃ 7) Elemental analysis value') Theoretical value as C2i$H54o4 C, 67,95; H, 8,43 Experimental value C white 8
．． 05: H, 8,378) TLC TLC grade: Silkal Art5715 manufactured by Merck & Co., Ltd. Solvent: Benzene: Seven tons = 1:1 Town value 0.64 (3) Wash the extract of (1) with saturated saline Then, by dissolving the mixture into the aqueous layer using a 5tlI sodium carbhydrogen aqueous solution, a fraction containing 9 M-4'cal d1711 sodium salt was obtained. This water layer is 0. The concentration was adjusted to -8.0 with IN hydrochloric acid, and then the fraction containing M-4' carline acid sodium salt was adsorbed on a Diaion HP-20 column (manufactured by Mitsubishi Chemical Industries, Ltd.). After eluting M-4' cardoic acid sodium salt with acetone and distilling off the acetone, it was freeze-dried to give M-4' sodium M7.
141 Jv of eJR sodium salt was obtained.

Physical properties of M-4' carmenic acid sodium salt 1) NMR spectrum: In heavy methanol, using TM8f as internal standard, 60
Measured with MEz. (CD, 00, a: pp
m ) 5.50 (1B, broad singlet) 5.70 (11, broad singlet) 5.95. (IH, quartet) 6.00 (IH, double fi 2) ultraviolet absorption spectrum (methanol solution) λmax
(am): 230 a 238 * 2463) Infrared absorption (KBr method) m-': 3400.
2900.1580 4) Elemental analysis i (external) Theoretical value as C2, H, 50, Nl C, 61,88:)l, 7.85 Experimental value C, 61,85;) 1,7.95 (4) The extract of (1) was washed with saturated saline, then an ether solution of diazomethane was added, and after standing for 30 minutes, it was dried under reduced pressure.
Then, using the resulting residue tube-par column (manufactured by Merck & Co., Ltd., 8160 size A), 7 tons of benzennia =
Purification with a 1=1 thread gave purified product 150119 of M-4' cal:tonic acid methyl ester as a colorless oil.

Physical properties of M-4'cal d'7flll methyl ester 1
) NMil spectrum - in deuterated chloroform, using TM8 as internal standard, 60
klnis text fl=. (cDct3#a
: ppm) 3.70 (3B, - heavy l1I) 5.50 (111, f load single aml) 5.75
(1B, )0-t(D-heavy 11i')5.90(1
B, quartet) 6.01 (IH*21 line) 2) Ultraviolet absorption spectrum (methanol solution) λwax
(am): 230, 238, 2463) External absorption spectrum (thin film method) m-': 3400.173
0 4) Mass spectrum N, O-hyx (T!J methylsilyl) After silylation with trifluoroacetamide, JEOL D-3100
Measurements were made using a

m/*: 654 (M”) 5) Elemental analysis value (ts) 024H580, theoretical value C, 65,73; H, 8,73 experimental value C, 65,6
6:H, 8,79 Production Example 2 M-4 type (1) Absidia coerulea I in 20 500-Yosaka flasks containing 100 mg of the culture medium with the following composition.
F04423 was inoculated at 1-26°C for 120 s, p, m.

(After 12 days of shaking culture at (stroke@s p*v m1nute)) Add ML-236B calcium phosphate sodium salt so that the final S degree is O, OS-, and culture at 26C, 120s, p for another 5 days. Cultured in m.

Medium composition Glucose 2.0% to 2HP840
．． 15 MgBsc7H200, 15 NH4No, 0.1 peptone
0.1 C,S-L O・2 Yeast extract 0. IZnB04'
7H200,001 Tap water Residue (pF (at 7.0)) After completion of the culture, the conversion reaction solution was filtered, and the F solution was adjusted to pH 3 with trifluoroacetic acid. Then, it was diluted with 1j of ethyl acetate three times. Upon extraction, a fraction containing M-4 carmenic acid was obtained.

Physical properties of M-4 carbic acid 1) TLC TLC plate: Merck Silica Rlum RT5715 Solvent: Benzene: Acetone: Vinegar 11 = 50:50:3R
f value 0.45 (2) The extract from (1) was washed with saturated saline, dehydrated with sodium sulfate, and then lacto/converted by adding a catalytic amount of trifluoroacetic acid to 5 bags of sodium bicarbonate. After washing with , it was dehydrated with sodium sulfate and concentrated to dryness to obtain lactone class 1i-. The C was applied to a Roper column (Merck G, 5iso size) and eluted with ethyl acetate to separate and collect the M-4 lactone fraction. This was further eluted using a Roper column (manufactured by Merck & Co., RP-8 size A) with 35% acetonitrile to prepare N.

Physical properties of M-4 lactone 1) NMR spectrum In deuterated chloroform, using TM8 as an internal standard, 10
Measured at 0 MHz. (cDct3.δ:ppm)4
．． 38 (1B, multiplex 11) 4.41 (1B, #) 4.62 (IH, y) 5.41 (IB, #) 5.58 (IH, z) 5.90 (IH, quartet) 6 .01 (IH, double line) 2) Ultraviolet absorption spectrum (methanol solution) λwax
(am): 230 * 236.7 t 244.
63) External absorption spectrum (thin film method) cx-': 34
00.2950.1725 (3) The extract obtained in (1) was washed with a saturated saline solution, an ether solution of noadimethane was added, and after standing for 30 minutes, it was dried under reduced pressure. The residue was collected using a Roper column (manufactured by Merck & Co., Ltd., 81
60 size A), penzene:ethyl acetate=1:1
The fraction containing M-4 methyl ester was collected. This was added to a Roper column (manufactured by Merck & Co., Ltd., BP-8).
Using size A), elute with 35% acetonitrile, -
After purification, purified sample 20 of M-4 Calgon methyl ester was obtained as a colorless oil.

Physical properties of M-4 calgenic acid methyl ester 1) NMR spectrum weight tz*200 using TMS for internal standardization in form
Measured at 1 GIm. (CDCts, δ: ppm)
0.88 (3H, t, J=7.3Hz) 0.8
9 (3B, d, J=6.5Hz) 1.12
(3B, d, J=6.8Hz) 1.1 to 1.7 (101
1, m) 2.34 (I H, sex, J-7Hz)2.
3-2.5 (2H, m) 2.49 (2B, d, J-6,4H$)2
．． 58 (III, va) 3.72 (3B, s) 3.78 (IB, m) 4.25 (1) 1, quin, J==7Hz
)4.4 (1B, m) 5.42 (IH, m) 5.56 (IH, m) 5.90 (IH, d, d, J=9.8, 5.6Hz
)5.99 (I H, d, J=9.8Hz)2
) Mass spectrum N, After silylation with 0-bis(trimethylsilyl)trifluoroacetamide, measurement was performed using JEOL D-300 model.

Sound: 654 (M”), 552,462.372,29
0°272.233,231°3) Ultraviolet absorption spectrum (ethanol solution) λmax
(am): 230.1, 237.3, 246.
44) Infrared absorption spectrum (thin film method) 11:3400
．． 2950.1730 5) Ding LC Ding LC Delay): Silica Germ rt5715 manufactured by Merck & Co. Solvent; Benzene: Acetone = 1:1 Town value 0.88 (4) 100 Da of the M-4 lactone obtained in (2) Dissolved in a small amount of acetone, added an equivalent amount of sodium hydroxide and left at room temperature for 1 hour. After indwelling acetone in a container, the product was freeze-dried to yield about 105 μm of M-4 carboxylic acid sodium salt.

Physical properties of M-4 Calginate sodium salt 1) NMR spectrum in heavy methanol, 1% birch standard: 200% using TMS
Measured in MBm. (CDsOD, δ: 1 house) 0
．． 91 (3B, t, J=74H vine) 0.92
(3B, d, J='1lix)1.12 (
3B, d, Jw7Hm) 1.1-1.8 (I
OH, m) 2, 25 (I H-d -d a J ”” 15
p 1-6 Hz ) 2.34 (11, d, d, J=1
5.5.5Bm) 2.2-2.4 (3B, m) 2.4B (IH, m) 3.6B (1B, m) 4.07 (1B, m) 4.28 (IH, m) 5.36 (1B, m) S-48 (1B, d, d, J = 3.2Hz) 5.88 (
1B, d, d, J = 9.6.5.3Hz) 5.98 (I
n, d, J = 9.8Hz) 2) Ultraviolet absorption spectrum (
methanol solution) λwax (11m): 230.0
, 237.2, 245.03) Infrared absorption spectrum (KBr method) m-': 3400.2900, 172
5.15804) TLC TLC plate; Silkal Art5715 manufactured by Merck & Co. Solvent; 7 tons of benzene: acetic acid = 50:50:3nf
Value 0.45 Production Example 36-Hydroxyiso ML-236B Fist (1)
By culturing in (1) of Production Example 2, M-4 was added to the extract.
6-Hydroxyiso ML-236B with Carzanoic Acid
A section containing cargenic acid was obtained, and the Rf value of 6-hydroxyisoML-236B cargenic acid was the same as that of M-4 carboxylic acid.

(2) In the treatment of Production Example 2 (2), after lactonization with trifluoroacetic acid, use a Roper column (Merck Co., Ltd. 1F).
The 6-hydroxyisoML-2368 lactone fraction was separated and collected during elution with ethyl acetate. This was purified in the same manner to obtain 6-hydroxyiso ML-236B lactone purified specimen 82
was gotten.

Physical properties of 6-hydroxyiso ML-236B lactone 1) NMR spectrum 1 In chloroform, using 7M81 as an internal standard.
The NMR spectrum measured at 0.00 MHz is shown in FIG.

2) Ultraviolet absorption spectrum (methanol solution) λvna
x (am): 229 r 234.8 + 244.
53) Infrared absorption (C77L'): Shown in the second fat.

4) Ding LC T LCf rate: Merck & Co., Ltd. Silical r Art 57
15 Solvent; be/zeniacetone: acetic acid = 50:50
: 3 town value 0.62 (3) In the treatment of (3) of Production Example 2, after alkylation with diazomethane, Roper column (manufactured by Merck & Co., Ltd.,
8160 size A,), benzene: When eluting with an ethyl acetate thread, 6-hydro90x-iso ML-23
6B carmenic acid methyl ester was separated and collected. This was purified in the same way and 6-human roxyine ML-2368
Purified sample 78119 of cargenic acid methyl ester was obtained.

Physical properties of 6-hydroxyisoML-236B carboxylic acid methyl ester 1) NMR spectrum In deuterated chloroform, using TMS as an internal standard,
The NMR spectrum measured at 0MH3 is shown in FIG.

2) After silylation with mass spectrum Ne0-1'su()su,tylsilyl)trifluoroacetamide, JEOL aiD-300
Measured using a mold.

m/: 654 (M”), 552,462,372
, 272°233.231 - 3) Ultraviolet absorption spectrum (methanol solution) λmax
(am): 229, 234.8, 244.54
) Infrared absorption spectrum (thin film method) CF2: shown in FIG.

5) TLC TLC plate: Silical Art 57 manufactured by Merck & Co.
15 Solvent; Benzene: Acetone = l: I Rf value 0.88 (4) 6-Hydroxyiso ML obtained in (2)
100 jv of -236B lactone was dissolved in a small amount of acetate, an equivalent amount of sodium hydroxide was added, and the mixture was left at room temperature for 1 hour. Then, after distilling off the acetone, the frozen 11L was dried to give 6-hydroxyino ML-236B cal&y11.
! About 103 da of sodium salt was obtained.Physical properties of 6-hydroxyiso ML-236Bcalz7#1 sodium salt 1) Ultraviolet absorption spectrum (methanol solution) λn;H
, (+m): 229 (sh) + 235, 2
45 (@h)2) Infrared absorption spectrum (KBr method)
cIL-': 340G, 285G, 1710.1580
3) TeLC TeLC plate; Silica gel Art5715 manufactured by Merck; Solvent; Benzene: Acetone: Vinegar gll=50:50:3
Town value 0.45 Manufacturing example 46-Medoxyin ML-236B class (1)
Apsinoa coerulea IFO44 was added to 20 500-capacity Erlenmeyer flasks containing 100 dt of a medium with the following composition.
23 was inoculated and cultured with shaking at 26°C and 220 r-pom-. After 4B, ML-2368 lactone was added to the final concentration of 0.051, and the cells were incubated for another 6 days at 26°C and 220 r-pom-.
Cultured at 20 r,p,m.

Medium composition glucose 2.0 2HP840
．． 15 Mg804・7B20 0.15NH4N
0. 0.1 peptone
0.1 C, 8, L O, 2 Yeast Extract 0.1” SOa ・17H200,
001 Tap water residue (pH 7, oKlmtF) After completion of the culture, the conversion reaction solution was filtered, and the FM was adjusted to -3 with trifluoroacetic acid. Then, with lj of ethyl acetate, 3
When extracted twice, 6-Medoxyiso ML-236B Calgan#! A classification including The above extract was washed with saturated saline, dehydrated with anhydrous am sodium, and then lactonized by adding a catalytic amount of trifluoroacetic acid.Then, the above extract was washed with 5 bags of sodium bicarbonate water bath solution, and anhydrous After dehydration with IJum (sulfuric acid), the mixture was dried under reduced pressure. *The sound object is a Roper column (manufactured by Merck & Co.).

Using 8i60t (Zum), benzene-acetone (7:
3) The 6-medoxyiso ML-236B lactone was collected by elution with the system. When this was purified, the target product 23 da was obtained.

Physical properties of 6-MedoxyisoML-236B lactone 1) NMR spectrum (coct3.δ: ppm) 90 MH in deuterated chloroform using 7M8 as an internal standard
Measured at z.

6.15 (1B, doublet) 5.70 (IB, quartet) 5.70 (IH, multiplet) 5.40 (1) 1. 1) 4.60 (IH, #) 4.40 (1B, #) 3.45 (IH, double line) 3.30 (3) 1. - Weight 1) 2) Ultraviolet absorption spectrum (methanol solution) λwax
(Field): 235 3) Infrared absorption spectrum (thin film method) CIL-34
50,1730 4) Mass spectrum: 420 (M”), 402#318,300,2
86゜68 5) TLC rate; Silicadal Art 57 manufactured by Merck & Co.
15 Solvent; Benzene: Acetone = 1:1 Town value 0.7 6) Elemental analysis value (excellent) CuHsbob theoretical value 0, 68.54:) I, 8.63 experimental value c
, 68.53: H, 8,81 (2) 6-Medoxyiso ML-236B lactone xtt - dissolved in a small amount of acetone, then 0.2N aqueous sodium hydroxide solution 131
After the completion of the 6 reaction of adding and hydrolyzing at 40°C for 1 hour,
Acetone was distilled off from the reaction mixture, and then washed with chloroform. water layer to 0. -8.0 gl with IN hydrochloric acid
It was adsorbed on a Hp-zo column (product of Ryo Kasei Kogyo Co., Ltd.).The fraction containing 6-Medoxyin ML-236B Carmen Shun sodium salt was eluted with 50 ml of acetone. After the acecin was distilled off, the product was freeze-dried to obtain 6-Me4:/ML-236B (My@sodium salt 1.003).

Physical properties of 6-Medoxyiso ML-236B carlinic acid sodium salt 1) NMR spectrum-1 In methanol, using 7M8t as an internal standard, 6
Measured at 0 MHz. (CD, OD, a:p
pm ) 3.31 (3) 1. -weight 1M) 5.42 (1) 1. Multiple + 131) 5.70 (11, Multiple #), 5.71 (1) 1. Quadruple-) 6.12 (1B, doublet) 2) Ultraviolet absorption spectrum (methanol solution) λwax
(Heavy tm) = 235 3) Infrared absorption spectrum (thin film method) 11: '3400
,2950,1580 4) Elemental analysis value (%) C24H570, theoretical value as Na CI 62.61; H, 8,04 experimental value C
, 62, 54: H, 8° 11, (3) 6-Medoxyiso ML-236B Cal-M/acid sodium salt 1? was dissolved in a small amount of methanol, and then trifluoroacetic acid was added under cooling to make it acidic. A diazomethane solution was immediately added and the mixture was left to stand for 30 minutes. After the reaction was completed, the solvent was distilled off from the reaction mixture. The obtained residue was purified using a Roper column (manufactured by Merck & Co., RP-8 size B) with a thread of methanol:water = 6 = 4 to obtain 6-medoxyiso ML-2.
36B cal&y acid methyl ester 78011kfI was obtained as a colorless oil.

Physical properties of 6-Medoxyiso ML-236B cargenic acid methyl ester 1) NMR spectrum in deuterated chloroform, using TM8t- as an internal standard,
Measured at 0 MHz. (CD, CL, δ: ppm)
3.30 (3H, -1+1l) 3.60 (3B, - double line) 4.37 (IEl, multi-lt line) 4.62 (IH, multi-ml line) 5.30 (IH, quadruple line) 5 .70 (IH, multi-line 1 line) 6.13 (IH, double III) 2) Ultraviolet absorption spectrum (methanol solution) λwax
(am) 2 235 3) External absorption spectrum (thin film method) 11:3400.
1725 4) Mass spectrum N, measured using JEOL D-300 after silylation with 0-bis(trimethylsilyl)trifluoroacetamide m/, : 668 (M+) 5) Elemental analysis value (% ) C25H400, theoretical value c, 66.34; H, 8,91 experimental value C, 66,
38; H, 8,98 Example 53-Hydroxy-MB-
Class 530B (1) 20 500-capacity Erlenmeyer flasks containing 100-mL culture medium with the following composition were inoculated with Mucor hyimaris IFO5834.
Shaking culture at 6°C, 220 rpm, 4 days later -M
B-530B lactone was added to a final concentration of over 0.05, and the mixture was further incubated at 26°C, 220r, p, m for 6 days.
・Cultivated in

Medium composition Glucose 1.0 PET 7 0.2 Meat extract 0.1 Yeast extract 0.1 Corn steep liquor '0.3 Tap water
Residue (pH not corrected) After completion of the culture, the conversion reaction solution was filtered and adsorbed onto a column using P'1rLt-Diaion 11P-20 (manufactured by Mitsubishi Chemical Industries, Ltd.). Next, elution was carried out with 70% methanol, and the eluate was distilled off. Then, the concentration was adjusted to -3.0 with trifluoroacetic acid. Then, extraction twice with vinegar 11 ethyl 1j yields 3-hydroxy-MB-530B Cal &
A classification containing 7 was obtained. 3-Hydroxy-MB-5
5 Solvent; 7 tons of benzene: acetic acid = 50:50:3 Town value 0.47 (2) An ether solution of diazomethane was added to the extract obtained in (1) and left for 30 minutes. The extract was then washed with saturated brine, and then dried under reduced pressure.

The obtained residue was applied to a microdandanotub Cl8 (manufactured by Waters), eluted with 53e6 methanol,
The fraction containing 3-hydroxy-MB-530B carmenic acid methyl ester was separated and collected.

This was purified to obtain 180 da of the target compound.

Physical properties of 3-hydroxy-MB-530B cardic acid methyl ester l) NMR spectrum in deuterated chloroform, using TM8t as internal standard, 9
Measured at 0MHz. (cDct, , δ:ppm)3
．． 72 (3H, - double line) 4.28 (II (, single M)' 5.45 (2H, multiple X #1) 5.93 (1B, quadruple sparse) 6.01 (II (, double 311 IM) ) 2) Ultraviolet absorption spectrum (methanol solution) λmax
(am): 229 + 236 e 244.53
) Infrared absorption spectrum (thin film method) (17K.

3410.2975.1730 4) Elemental analysis value (theoretical value as phantom C25H4゜0.
C, 66,34; H, 8,91 Experimental value C, 66,30
;H,9,02(3)3-hydroxy-MB-530B
Powerful? 100'mg of acid methyl ester was added to 0. It was dissolved in IN aqueous sodium hydroxide solution and stirred at 30°C for 1 hour.

The solution was then washed with chloroform and lyophilized to yield 3-hydroxy-MB-530B cal-I acid sodium salt 93Wv.

Physical properties of 3-hydroxy-MB-530B carmenic acid sodium salt 1) NMR spectrum Measured at 90 MHz in deuterated water using 088 as internal standard * (D20 * δ; ppm) 5.40 (2
H, multiplet) 5.89 (1B, quadruplet) 6.00 (IH, doublet) 2) Ultraviolet absorption spectrum (aqueous solution) λwax (am): 229, 236, 2453
) Infrared absorption spectrum (KBr method) cIL.

3400 - 2900.1580 4) Elemental analysis value C1) C2aHsy07'' Theoretical value C, 62,59; H, 8,097 Experimental value C, 6
2,37:H,8,21 Production Example 66-HydroxyisoMB-530B (1) By culturing in Production Example 5 (1), the extract contains 3-hydroxy-=MB-530B calgenic acid and 6 -Hydroxyin MB -530B A section containing cardanic acid was obtained, and 6-Hydroxyiso MB
-530B Cal-17 acid Bf value is 3-hydroxy-MB
It was the same as -530B Carboyli.

(2) After culturing in Production Example 5 (1), the extract was washed with saturated saline, dried over sodium &/L acid, and then a catalytic amount of trifluoroacetic acid was added. Next, extract f: 5 bags were washed with sodium hydrogen oxide, dried with sodium sulfate,
It was dried under reduced pressure. The obtained residue was eluted with ethyl acetate using a Roper column (manufactured by Merck & Co., Ltd., 5i60 size A), and the 6-hydroxyisoMB-530B lactone was separated and collected. This was purified to obtain the target compound 212.

Physical properties of 6-hydroxyloxyine MB-5308 lactone 1) NMB spectrum 90 M in deuterated acetone using TMS as an internal standard.
Measured with H&. (CD, COCD, , δ: ppm
)3.94(1)1. 2M [gun] 4.35 (1) 1. Multiplex 48ri 4.66 (IH, multiplex) 5.45 (1) 1. Multi-X-ray) 5.62 (1B, 211L Aya) 5.91 (188-mu) 2) Ultraviolet absorption spectrum (methanol solution) λrr+
ax (nm): 238.43) Infrared absorption spectrum (KBr method) art-': 3450.1730 4) Mass spectrum rv,: 420 (M") 5) Elemental analysis value (theoretical value as C24H5606)
C, 68,54: H, 8, 63 Experimental value C, 68, 33
: H, 8, 81 (3) 6- Dissolve 100 Da of human rxjt Seaia MB-530B5 chtonic body in a small amount of acetone, then hydroxylate an equivalent amount of 0.2N) in a water bath wLt- and at room temperature. Stirred. This solution was then subjected to lyophilization to obtain 6-hydroxyisoMB-530.
1061v of B carmenic acid sodium salt were obtained.

Physical properties of 6-hydroxyisoMB-530B carboxylic acid sodium salt 1) NMR spectrum In deuterated water, using D88t- as an internal standard, 90M) is
It was measured with (D20 +δ': ppm) 3.7B
(1B, multiple fL4!i-ri 4.01 (1B, multiple l1I) 4.13 (IH, multiple *) 5.49 (1B, multiple line) 5.62 (1) 1.multi] [lti-ri 6. 02 (IH, wide singlet #) 2) Ultraviolet absorption spectrum (aqueous solution) λmax (theory m): 238 3) Infrared absorption spectrum (KBr method) rx-':
3400-2900.1575 4) Elemental analysis value C%) Theoretical value as C24HsyOyNa C, 62,59; H, 8,097 Experimental value C,
62,28:H, 8,31 (4) Production Example 5 (2)
In the treatment, after alkylation with noadimethane, it was subjected to a micro dt7 dapatsu"18, and the wrinkles were removed with 53 tatami methanol, and the fraction containing 6-, dolo, and xyiso MB-5308 carmen kin methyl ester was separated and collected. This was purified. 170 da of the target compound was obtained.

Physical properties of 6-hydroxyloxyiso MB-530B cal-7 acid methyl ester 1) NMR spectrum Measured at 9cm in deuterated acetone using TM8 as an internal standard L7'j. (CD, COCD, , δ: ppm
) 3.70 (3) 1 - multiple edges) 4.22 (IH, multiple weight) 5.42 (IH, multiple weight > 5.57 (IH, multiple lines) 5.92 (18, double #) 2 ) Ultraviolet absorption spectrum (methanol solution) λmax
(m): 238.4 3) Infrared absorption spectrum (thin film method) cix-'23400.2960.1732 4) Elemental analysis value (%) Theoretical value as C2584007 C, 66,34: H, 8,91 lol Experimental value c, 6
6.28 ; H, 8,99 The ~ bile acid absorbent used in the composition of the present invention is a pharmaceutically acceptable basic anion exchange resin that binds bile acids in the gastrointestinal system and absorbs them in the intestine. It is excreted in feces and cannot be reabsorbed. Suitable basic ion exchange resins are as follows.

The first type is a copolymer of styrene and novinylbenzene into which a basic group is introduced.

For example, cholestyramine contains a quaternary ammonium functionality attached to a styrene-novinylbenzene copolymer. The main constituent is polystyrene trimethylbennolammonium present with Ct-anions, but also contains novinylbenzene (2) and water (0-75). Degree of crosslinking: 1-10° 111A group of main polymer: It has a structure represented by the following, and is preferably 50-100 mesh. This is described in US Pat. Nos. 3,383,821 and 3,308,020.

acetic acid, polyalkylene polyamines and epichlorohydrin, glycerol-1,3-dichlorohydrin or aliphatic bisepoxy compounds (1゜2:3,4-epoxybutane, bisepoxyglobin ether, or alpha, omega) - Compounds such as alkylene glycols). For example, cholesbitol is a copolymer of diethylenetriamine and 1-chloro-2,3-epoxygulonone, and is a crosslinked polymer of these two substances. It is obtained by bringing these two substances to about 4 with hydrochloric acid, then dialyzing and drying the dialyzed material.

In the reaction product, the amino group portion is quaternized to form a hydrochloride.

The third type is the cross-linked polysaccharides omega-noalkylakinoalkyl, omega-di(hydroxyalkyl)-aminoalkyl and omegamorpholinoalkyl ethers, such as dextran, starch and saccharide, which contain acids. It's like cross-linking with Drino. For example, dextran is added to the diethylaminoethyl 5-halodanate in the presence of a water-soluble alkali such as concentrated sodium hydroxide.
After treatment with cloud and crosslinking with shrimp chlorohydrin, the product is poly[2-(diethylamino)-ethylcopolyglycerylene dextran. Such compounds are described in US Pat. No. 3,277,025.

The fourth button is made of quaternary ammonium anion hybrid resin.
The preferred one is designated as poly-[methyl-(3-)IJ methylammoniopropyl)imi□2trimethylene-tubaride] and has the following structure.

Such compounds are described in US Pat. No. 4,098,726.

Basic, non-toxic, ion exchange resins, such as resin, are water-soluble or water-insoluble or swell in the presence of water. These trees are preferably used in monosalt form, for example with non-toxic salts such as hydrochloric acid, phosphorous acid or phosphoric acid.
Preferably used as brewing salt, the resinous material is used in the form of fine granules.

The compositions of the invention are co-administered with the active ingredient in separate dosage forms and preferably in combination for the treatment of patients suffering from hypercholesterolemia.
For compounds, 005-25-/V days, preferably Fi, 0.2
~20~/hour 7 days, M~4' compound and 6-
Hydroxyin ML-236B 1ml compound is 0.
1 to 50 da//day, preferably 0.5 to 25 to 9/day
Sunflower, 3-hydroxy-MB-530Blil
compound and 6-hydroxylisoMB-530B class compound, 0.05 to 25 ttv/#/day, preferably 0
, 2 to 209/day/day, and 6-MedoxyisoML
-236B class compounds: 0.05 to 5 ow/kll/day, preferably 0.5 to 30 ow/kll/day, preferably 0.5 to 30 ow/kll/day.

The amount of cholestyramine used is 3.5 to 2117 days, preferably Fi12-15) 7 days. For infants and children, a dosage of 0.3-1.1f/7 days is preferable, taking into account their body weight.The composition of the present invention can be administered orally in the form of tablets, capsules, etc. Oral administration is usually preferred, and the dosage form can be varied depending on the age, symptoms, weight, and other conditions of the patient, but the daily dosage for adults is within the range of 10 to 100 ay. It will be given in four parts. Larger doses can be administered as needed.

Next, an example will be shown in which the above-mentioned cholesterol synthesis inhibitor and cholestyramine were administered to Peagle-sized animals.

The experiment used Peagle-sized animals weighing 7-10 kilograms (average 8.5 kg). Blood was collected once a week for 3-4 days before administration, and the average blood cholesterol level was taken as the pre-administration value. Blood is drawn once a week and blood cholesterol'it:
ta fixed shi'tc.

Example I Reduction of blood cholesterol in dogs by administration of M-4 carboxylic sodium salt and cholestyramine (4) group represents control. During the 4th week of the m phase (1), the ω) group received only M-4 carvone sodium salt, and the ω) group received only cholestyramine - late stage 4.
Week ω], M-4 carboxylic acid sodium salt + cholestyramine was administered to both groups (■). The dosage is M-4 carmen sodium salt once a day in a gelatin capsule at 20η/dark/day, and cholestyramine mixed in food at 11gl/day. /YU was administered for 7 days.

The results are shown in Table 1.

26163 24 3 'a IOL, 7 56 6
163 32 68 (56)7
♀ 104 36 77 (58) lO
♂ 124 35 71 (57) 11
♀ 157 35 68 (57
) Book φ) Expected value of group = pb + PC” (100-Pb)
00 Pb: Decrease rate in (BJ group 0), PC: (C dust (average decrease rate in IJ (35%)) In the first 4 weeks [period (■)],] M-4 sodium carphosphate Blood cholesterol decreased by an average of 34 qb in the group B) to which only cholestyramine was administered, and by 35 qb in the group C) to which only cholestyramine was administered. This decrease occurred after administration 2.
The maximum value was reached after -3 weeks, and no further decrease in p was observed.

During the next four weeks [Period 1], cholestyramine was administered to the ψ) group, and M-4 carlinic acid sodium salt was administered to the 0 group in combination, respectively. For group (B), the expected reduction in blood cholesterol by administering cholestyramine in combination, 1il (shown as the expected value in the table)
The average score is 57. On the other hand, the actual average is 741
However, the combination of cholestyramine exceeded the expected value. Similarly, in the case of group (), M-4
Compared to the expected average reduction in blood cholesterol of 57 or more by administering sodium carlinic acid salt in combination, a reduction of 70% was actually observed.

Example 2. Reduction of blood cholesterol in dogs by administering 6-hydroxyloxyiso ML-236B carmenic acid sodium salt and cholestyramine In Example 1, 6-hydroxyloxyisomer was used instead of M-4 carboxylic acid sodium salt. The test was carried out in the same manner as in Example 1 except that ML-236B carmenic acid sodium salt was used.

The results are shown in Table 2.

Table 2 2613634 3 9 139 2 3 6 ♂ 143 24 62 (48)
7 ♀ 102 27 66 (50)
10 a 147
32 65 (50) 11 ♀
112 34 67 (52) Book The method of calculating the predicted value is the same as described above.

In period (1), 6-hydroxyisoML-236B
Group B, which received only sodium calphosphate, had an average of 27 cases, and group B, which had only cholestyramine, had an average of 3 cases.
2 winners, each with a decrease in blood cholesterol. period(
II) in butterflies, (in group 83 there was an average reduction of 65%, which exceeded the average reduction of 50% expected by combination administration, (
In group C), the average expected decline rate was 6 compared to the expected average of 52.
A decrease of 6 points was observed.

Example 3. Reduction of blood cholesterol levels in dogs by administration of 3-hydroxy-MB-530B calgenic acid sodium salt and cholestyramine In Example 1, 3-hydroxy-MB was added to the M-4 calgenic acid sodium salt. -530B Kargen Shunsu)
The test was conducted in the same manner as in Example 1 except that lithium salt was used.

The results are shown in Table 3.

Table 3 (AJ1614253 2♂113 0 4 3♀128 3 4 49124 52 Average 127 3 3 (8) 5
6 114 33 68 (56
) 68108 36 78 (57) 7♀131 33 72 (56) 8♀122 35 75 (58) Average 119 34 73 (57) (
C) 96126 36 73 (58) 108131
34 73 (56) 119107 35 72 (57) 129125 35 70 (,57) * The calculation method for pre-m4iILo is the same as above.

In period (1), only a-humanoxy-MB-530B Calme 1ill sodium salt was administered. ω) # was 34 on average.
Group C), which received only cholestyramine, had an average of 35%
, blood cholesterol decreased respectively. Period (U)
So, in group (6), the average reduction rate exceeded the expected average reduction rate of 57% due to combination administration, and in group (C), the average reduction rate was 72. Admitted.

[Brief explanation of drawings]

Figure 1 shows 6-hydroxyisoML-236B-)
Figure 2 shows the magnetic resonance spectrum of the chthonic body, and Figure 2 shows the infrared absorption spectrum of the same substance. FIG. 3 shows the nuclear magnetic resonance spectrum of 6-hydroxyiso ML-2368 carinic acid methyl ester, and the 4th rg shows the infrared absorption spectrum of the same substance. Patent Applicant Sankyo Co., Ltd. Representative Patent Attorney Izuru Kashi Procedural Amendment (Voluntary) October 2, 1975/:) Japan Patent Office Commissioner Kazuo Wakasugi 1, Indication of Case 1988 Patent Application No. 188530 No. 2, Name of the invention Composition for the treatment of hypercholesterolemia 3, Relationship with the amended person's case Patent applicant address 6 Name, 3-1 Nihonbashi Honmachi, Chuo-ku, Tokyo 103 (185) Representative of Sankyo Co., Ltd. Person: Director and President Yoshinori Kawamura 4, Agent Address: 7, Sankyo Co., Ltd., 1-2-58 Hiromachi, Honbunagawa-ku, Tokyo, 140 Contents of amendment: Annex 1, page 4, lines 8 and 11 of the specification r4111Ws
5! -124315” to “Unexamined Japanese Patent Publication No. 5T-501194
Correct it to "No.". 1 "% Kaikai 57-1" of "Special Application S!1-t15483" on lines 16 and 17 of the same page
No. 01031”. L Correct “p・ma” on page 1, line 1 of the same page to “p・r”. 4 On page 21, line 14, [OD, 07 J is corrected to "0DCts". i Correct "Magnetic Resonance Spectrum J" on page 50, line 12 to read "Nuclear Magnetic Resonance Spectrum."that's all

Claims (1)

[Claims] 1. a) having the formula (wherein R#i represents a hydrogen atom or a methyl group, V represents a hydrogen atom or a methyl group, and Rs represents a hydroxyl group or a methoxy group) CaldfVW1%7O, a pharmacologically acceptable salt thereof, a lower alkyl ester thereof, or a cholesterol synthesis inhibitor thereof, and b) a bile acid that binds to bile acids in the gastrointestinal system and makes reabsorption impossible. 1. A composition for treating hypercholesterolemia, comprising a pharmaceutically acceptable non-abrasive basic anion conversion resin.