GB2073199A - Compounds Which Inhibit Cholesterol Biosynthesis, and Their Preparation - Google Patents

Abstract

Compounds of formula (I): <IMAGE> (wherein R<1> represents a hydrogen atom or a methyl group; R<2> represents a hydrogen atom, the alcoholic moiety of an ester or the cationic moiety of a salt; R<3>, R<4> and R<5> are the same or different and each represents a hydrogen atom or an organic or inorganic acyl group, provided that, when R<3> represents a 2-methylbutyryl group, R<4> and R<5> both represent acyl groups; and n is the valency of R<2>) may be prepared by cultivating microorganisms of the genera Penicillium or Monascus or by salifying or esterifying the corresponding carboxylic acid or lactone or, in the case of esters of the above formula (I), by esterifying a corresponding salt of the above formula (I).

Description

SPECIFICATION Compounds Which Inhibit Cholesterol Biosynthesis, and Their Preparation The present invention relates to a series of novel compounds which have been found to inhibit cholesterol biosynthesis and which can thus be used in the treatment and prevention of disorders arising from high levels of cholesterol in the body. The invention also provides processes for preparing these compounds.

Hyperlipaemia, especially hypercholesteraemia, is known to be one of the main causes of cardiopathy, such as cardiac infarction or arteriosclerosis. As a result, considerable research has been carried out in an effort to discover compounds capable of reducing lipid, and especially cholesterol, levels in the blood. A group of compounds of this type is disclosed in US Patent Specification No.

3,983,1 40 and was isolated from microorganisms of the genus Penicillium; this group of compounds is collectively designated ML-236. Also, United Kingdom Patent Applications No. 2046737 and No.

2049664 disclose a structurally similar compound designated Monacolin K or MB-530B, and certain salts and esters of MB-530B are disclosed in United Kingdom Patent Application No. 2055100.

MB-530B and its salts may be prepared by cultivating microorganisms of the genus Monascus, especially, but not exclusively, strains of Monascus ruber. Another compound of the MB-530 group, designated MB-530A, as well as other compounds related to the ML-236 and MB-530 groups are disclosed in our co-pending United Kingdom Patent Application No. 8110075.

We have now discovered a series of compounds related to the ML-236 and MB-530 groups which also have valuable inhibitory activity against the biosynthesis of cholesterol.

The compounds of the invention have the formula (I):

wherein: R1 represents a hydrogen atom or a methyl group; R2 represents a hydrogen atom, the alcoholic moiety of an ester or the cationic moiety of a salt; R3, R4 and R5 are the same or different and each represents a hydrogen atom or an organic or inorganic acyl group, provided that, when R3 represents a 2-methylbutyryl group, R4 and R5 both represent acyl groups; and n is the valency of R2.

The invention also provides processes for preparing the compounds of the invention by fermentation employing microorganisms of the genus Penicillium or Monascus or by hydrolysis, salification or esterification reactions commencing with such fermentation products.

As the compounds of the invention are derivatives of the ML-236 and MB-530 group compounds in which the lactone ring has been broken, they are named as derivatives of ML-236 carboxylic acids or MB-530 carboxylic acids (i.e. the carboxylic acids obtained by hydrolysis of ML-236 or MB-530 compounds) and the numbering system is that shown on the following structure:

(in which R' and R3 are as defined above).

In the compounds of formula (I), where H2 represents the cationic moiety of a salt, it may be a metal atom, for example an alkali metal atom (e.g. sodium or potassium), an alkaline earth metal atom (e.g. calcium, magnesium or barium), a transition metal atom (e.g. iron, nickel or cobalt) or other metal atoms (e.g. aluminium, zinc or copper); in this case, n will be the well-known valency of these metals, in the examples given normally varying from 1 to 3. Alternatively, it may be an ammonium group or a substituted ammonium group (preferably an alkyl-substituted ammonium group), such as methylammonium, ethylammonium, isopropylammonium, dimethylammonium, diethylammonium, trimethylammonium, triethylammonium, tetramethylammonium, or dicyclohexylammonium; in this case, n will be 1.Another cationic moiety which may be represented by R2 is the salt-forming group derived from a basic amino acid, such as lysine, arginine or ornithine.

Alternatively, R2 may be the alcoholic moiety of an ester, in which case the value of n will depend upon the nature of the hydroxy compound from which R2 is derived; for example in the case of monoalcohols, n would be 1, in the case of glycols, n would be 2 and in the case of glycerol, n would be 3.

Where R2 represents a monovalent group, this is preferably an unsubstituted or substituted alkyl group, an unsubstituted or substituted aralkyl group or an unsubstituted or substituted phenacyl group.

Examples of alkyl groups which may be represented by R2 include straight and branched chain alkyl groups, preferably having from 1 to 8 carbon atoms, for example the methyl, ethyl, propyl, isopropyi, butyl, isobutyl, sec-butyl, t-butyl, pentyl, sec-pentyl, t-pentyl, isopentyl, neopentyl, hexyl, heptyl, 2-methylhexyl and octyl groups.

Examples of aralkyl groups which may be represented by R2 include the benzyl group and the benzhydryl group, both of which may be unsubstituted or have one or more substituents in the benzene ring. Examples of such substituents include: C1-C4 alkyl groups (methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyi or t-butyl), C,--C, alkoxy groups (methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy or t-butoxy), halogen atoms (e.g. chlorine, bromine or fluorine) or the trifluoromethyl group. Where there are two or more substituents, these may be the same or different.

Examples of such aralkyl groups include the benzyl, 2-methylbenzyl, 3-methylbenzyl, 4-methylbenzyl, 2-ethylbenzyl, 3-ethylbenzyl, 4-ethylbenzyl, 2-propylbenzyl, 3-propylbenzyl, 4-propylbenzyl, 2- butylbenzyl, 3-butylbenzyl, 4-butylbenzyl, 2-m ethoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl, 2- propoxybenzyl, 3-propoxybenzyl, 4-propoxybenzyl, 4-butoxybenzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4- chlorobenzyl, 2-bromobenzyl, 3-bromobenzyl, 4-bromobenzyl, 2-fluorobenzyl, 34luorobenzyl, 4- fluorobenzyl, 2-trifluoromethylbenzyl, 3-trifluoromethylbenzyl, 4-trifluoromethylbenzyl and benzhydryl groups.

Where R2 represents a phenacyl group, this may be unsubstituted or may have one or more substituents in the benzene ring. Examples of such substituents include: C1-C4 alkyl groups, C1-C4 alkoxy groups, halogen atoms and the trifluoromethyl group; specific examples of these substituent groups are given above.Preferred examples of such phenacyl groups include the phenacyl, 2 methylphenacyl, 3-methylphenacyl, 4-methyiphenacyl, 2-ethylphenacyl, 3-ethylphenacyl, 4- ethylphancyl, 2-propyiphenacyl, 3-propylphenacyl, 4-propyiphenacyl, 2-butylphenacyl, 3- butylphenacyl, 4-butylphenacyl, 2-methoxyphenacyl, 3-methoxyphenacyl, 4-methoxyphenacyl, 2- ethoxyphenacyl, 3-ethoxyphenacyl, 4-ethoxyphenacyl, 2-prnpoxyphenacyl, 3-propoxyphenacyl, 4- propoxyphenacyl, 2-butoxyphenacyl, 3-butoxyphenacyl, 4-butoxyphenacyl, 2-chlorophenacyl, 3- chlorophenacyl, 4-chlorophenacyl, 2-brnmophenacyl, 3-bromophenacyl, 4-bromophenacyl, 2- fluorophenacyl, 3-fluorophenacyl, 4-fluorophenacyl, 2trifluoromethylphenacyl, 3- trifluoromethylphenacyl, and 4-trifluoromethylphenacyl groups.

Where R2 represents a bivalent alcoholic moiety, it is preferably a C2-C6 alkylene or alkylidene group, for example an ethylene, ethylidene, propylene, propylidene, trimethylene, tetramethylene, butylidene, pentamethylene or pentylidene group, as well as such groups having one or more substituents, e.g. hydroxy groups, halogen atoms or trifluoromethyl groups.

Where R2 represents a trivalent alcoholic moiety, it is preferably a saturated aliphatic hydrocarbon group having from 2 to 6 carbon atoms and optionally one or more substituents, e.g. hydroxy groups, halogen atoms or trifluoromethyl groups.

Where R3, R4 or R5 represents an acyl group, this may be an organic acyl group (for example an aliphatic acyl group, an aromatic acyl group, an araliphatic acyl group, an alicyclic acyl group, a heterocyclic acyl group, a heterocyclic-substituted aliphatic acyl group, an aliphatic sulphonyl group, an aromatic sulphonyl group, an aliphatic phosphoryl group, an aromatic phosphoryl group or an araliphatic phosphoryl group) or an inorganic acyl group (for example an acyl group derived from phosphoric acid, sulphuric acid or nitric acid).

Where the acyl group is an aliphatic acyl group, this may be saturated or unsaturated and examples include: straight or branched chain C2-C20 alkanoyl groups (e.g. the acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, 2-methyivaleryl, heptanoyl, isoheptanoyl, octanoyl, isooctanoyl, 2-methyloctanoyl, nonanoyl, isononanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, palmitoyl, stearoyl, isostearoyl, nonadecanoyl and eicosanoyl groups);; C3-C20 alkenoyl groups (e.g. the acryloyl, crotonoyl, 3-butenoyl, methacryloyl, 3- methyl-2-butenoyl, 2-pentenoyl, 4-pentenoyl, tigloyl, angeloyl, 2-hexenoyl, 2-heptenoyl, hepta-2,4 dienoyl, 2-octenoyl, 2-nonenoyl, 2-decenoyl, 2-undecenoyl, linolenoyl, oleoyl, linoleoyl and arachidonoyl groups); and C3-C20 alkynoyl groups (e.g. the propioloyl, 2-butynoyl. 3-butynoyl, 2- pentynoyl, 2-hexynoyl, 2-heptynoyl, 2-octynoyl, 2-nonynoyl and 2-decynoyl groups).These acyl groups may be unsubstituted or may have one or more substituents, for example: halogen atoms, such as chlorine or bromine; the trifluoromethyl group; the nitro group; the carboxy group; alltoxycarbonyl groups, such as methoxycarbonyl or ethoxycarbonyl; aralkoxycarbonyl groups, such as benzyloxycarbonyl; the cyano group; the amino group; all < anoylamino groups; such as acetylamino; alkylamino groups, such as methylamino, dimethylamino or ethylamino; aralkylamino groups, such as benzylamino; the hydroxy group; alkanoyloxy groups, such as acetoxy or pivaloyloxy; alkoxy groups, such as methoxy or ethoxy; the sulphhydryl group; allcylthio groups, such as methylthio or ethylthio; and acylthio groups, such as acetylthio or benzylthio.

Where the acyl group represented by R3, H4 or R5 is an aromatic acyl group, the aromatic ring may be a single ring (e.g. a benzene ring) or a fused ring, for example a naphthalene ring, an anthracene ring or an indan ring. This ring may optionally have one or more substituents, for example alkyl groups, alkoxy groups, halogen atoms, trifluoromethyl groups, nitro groups, cyano groups, amino groups or hydroxy groups; where there are two or more substituents, these may be the same or different.Specific examples of such aromatic acyl groups include the benzoyl, o-toluoyl, rn-toluoyl, p-toluoyl, 2,4- dimethylbenzoyl, 3,4-dimethylbenzoyl, 2-ethylbenzoyl, 3-ethylbenzoyl, 4-ethylbenzoyl, 2- propylbenzoyl, 3-propylbenzoyl, 4-propylbenzoyl, 4-butylbenzoyl, o-anisoyl, m-anisoyl, p-anisoyl, 2,4- dimethoxybenzoyl, 2--ethoxybenzoyl, 3-ethoxybenzoyl, 4-ethoxybenzoyl, 2-propoxybenzoyl, 3- propoxybenzoyl,4-propoxybenzoyl,2-butoxybenzoyl, 3-butoxy benzoyl, 4-butoxybenzoyl, piperonyloyl, 2-chlorobenzoyl, 3-chlorobenzoyl, 4-chlorobenzoyl, 2,3-dichlornbenzoyl, 34-di6hlornbenzoyl, 2$4- dichlorobenzoyl, 2-bromobenzoyl, 3-bromobenzoyl, 4-bromobenzoyl, 24luorobenzoyl, 34luorobenzoyl, 4-fluorobenzoyl, 2-trifluoromethylbenzoyl, 3-trifluoromethylbenzoyl, 4-trifluoromethylbenzoyl, 2- nitrobenzoyl, 3-nitrobenzoyl, 4-nitrobenzoyl, 2,4-dinitrobenzoyl, 3,5-dinitrobenzoyl, salicyloyl, 3- hydroxybenzoyl, 4-hyd roxybenzoyl, 2-acetoxybenzoyl, 3-acetoxybenzoyl, 4-acetoxybenzoyl, anthraniloyl, 2-acetamidobenzoyl, 4-acetamidobenzoyl, vanilloyl, veratroyl, protocatechuoyl, galloyl, 1 - naphthoyl, 2-naphthoyl, 2-anthranoyl, 4-indanecarbonyl, 5-indanecarbonyl and 4-indenecarbonyl groups.

Where the acyl group represented by R3, R4 or R5 is an araliphatic acyl group, the aromatic ring in this araliphatic acyl group may be a single ring or a fused ring, as exemplified above for the aromatic acyl group, whilst the aliphatic moiety may be saturated or unsaturated. The aromatic ring may optionally have one or more substituents, as exemplified above for the aromatic acyl groups.Examples of such araliphatic acyl groups include the phenylacetyl, 2-methyiphenylacetyl, 3-methyiphenylacetyl, 4-methyiphenylacetyl, 2-ethylphenylacetyl, 2-methoxyphenylacetyl, 3-methoxyphenylacetyl, 4 methoxyphenylacetyl, 2-ethoxyphenylacetyl, 3-ethoxyphenylacetyl, 4ethoxyphenylacetyl, 2- propoxyphenylacetyl, 3-propoxyphenylacetyl, 4-propoxyphenylacetyl, 2,4-diniethoxyphenylacetyl, 3,4- methylenedioxyphenylacetyl, 2-hydroxyphenylacetyl, 3-hydroxyphenylacetyl, 4-hydroxyphenylacetyl, 2-chlorophenylacetyl, 3-chlorophenylacety, 4-chlorophenylacetyl, 2,3-dichiorophenylacetyl, 2,4dichlorophenylacetyl. 3,5-dichlorophenylacetyl, 2-bromophenylacetyl, 3-bromophenylacetyl, 4 bromophenylacetyl, 2-nitrophenylacetyl, 3-nltrophenylacetyl, 4-nitrophenylacetyl, 4-aminophenyl acetyl, 2-phenylpropionyl, 3-(2-methylphenyl)propionyl, 3-(3-methylphenyl)propionyl, 3-(4 methylphenyl)propionyl, 3-(2-methoxyphenyl)propionyl, 3-(3-methylphenyl)propionyl, 3-(4-methoxy phenyl)propionyl, 3-(3,4-methylenedioxyphenyl)propionyl, 3-(2-chlorophenyl)propionyl, 3-(3- chlorophenyl)propionvl, 3-(4-chlorophenyl)propionyl, phenoxyacetyl, cinnarnsyl, o-methylcinnamoyl, m-methylcinnamoyl, p-methylcinnamoyl, o-methoxycinnamoyl, m-methoxycinnamoyl, p methoxycinnamoyl, o-hydroxycinnamoyl, m-hydroxycinnamoyl, p-hydroxycinnamoyl, o- chlorocinnamoyl, m-chlorocinnamoyl, p-chlorocinnamoyl, o-bromocinnamoyl, m-bromocinnamoyl and p-bromocinnamoyl groups.

Where the acyl group represented by R3, R4, or R5 is an alicyclic acyl group, the alicyclic ring may be saturated or unsaturated and preferably has from 3 to 7 carbon atoms. Examples of such alicyclic acyl groups include the cyclopropanecarbonyl, cyclobutanecarbonyl, cyclobut-1-enecarbonyl, cyclobut2-enecarbonyl, cyclo-pentanecarbonyl, cyclopent-1 -enecarbonyl, cyclopent-2-enecarbonyl, cyclopenta-1,3-dienecarbonyl, cyclopenta-2,4-dienecarbonyl, cyclohexanecarbonyl, cyclohex-1 -ene- carbonyl, cyclohex-2-enecarbonyl, cyclohex-3-enecarbonyi, cyclohexa-1 3-dienecarbonyl, cyclohexa2,4-dienecarbonyl, cycloheptanecarbonyl, cyclohept- 1 -enecarbonyl, cyclohept-2-enecarbonyl, cyclohept-3-en ecarbonyl, cyclohepta-1 3-dienecarbonyl, cyclohepta-2,4-dienecarbonyl, cyclohepta2,5-dienecarbonyl, cyclohepta-1 4-dienecarbonyl, cyclohepta-1 ,5-dienecarbonyl, cyclohepta-1,3,5trienecarbonyl, cyclohepta-2,4,6-trienecarbonyl and adamantanecarbonyl groups. These rings may optionally have one or more substituents, as exemplified above in respect of the aromatic acyl groups and such substituents may form one or more rings fused to the abovementioned alicyclic ring system.

Where the acyl group represented by R3, R4 or H5 is a heterocyclic acyl group, the heterocyclic ring preferably has 5 or 6 ring atoms, one or more of which is a hetero atom, for example a nitrogen atom, an oxygen atom, a sulphur atom or a selenium atom. This heterocyclic ring may optionally be fused with a carbocyclic ring or with another heterocyclic ring (which may be the same as or different from the first-mentioned heterocyclic ring) to form a fused heterocyclic ring system. The heterocyclic acyl group may have one or more substituents, as exemplified above in respect of the aromatic acyl groups.Specific examples of such heterocyclic acyl groups include the 2-thenoyl, 3-thenoyl, 5 methylthen-2-oyl, 5-chlorothen-2-oyi, 4,5-dimethylthen-3-oyl, 2-furoyl, 3-furoyl, 5-methylfur-2-oyl, 5- chlorofur-2-oyl, pyridine-2-carbonyl, 3-methylpyridine-2-carbonyl, 4-methylpyridine-2-carbonyl, 5- methylpyridine-2-carbonyl, 6-methyl-pyridine-2-carbonyl, nicotinoyl, isonicotinoyl, isoxazole-3 carbon, isoxazole-4-carbonyl, oxazole-2-carbonyl, oxazole-4-carbonyl, 4-acetylaminothiazole-2- carbonyl, 1 ,3,4-thiadiazole-2-carbonyl, 5-methyl- 1 ,3,4-thiadiazole-2-carbonyl, 1 ,2,3-triazole-1 - carbonyl,1,2,3,4-tetrazole-1-carbonyl, piperidinecarbonyl, 4-methyl-1 -piperazine-carbonyl, 1 - pyrrolidinecarbonyl, benzofu ran-2-carbonyl and benzothiophene-2-carbonyl groups.

Where the acyl group represented by R3, R4 or R5 is a heterocyclic- substituted aliphatic acyl group, the heterocyclic group may be as described above for the heterocyclic acyl groups and the aliphatic moiety may be saturated or unsaturated and may be as described above for aliphatic acyl groups. Examples of such heterocyclic-substituted aliphatic acyl groups include the 2-thienylacetyl, (5 methylthiophene-2-yl)acetyl, (5-chlorothiophene-2-yl)acetyl, 3-thienylacetyl, 2-furylacetyl, 3- furylacetyl, 2-pyridylacetyl, 3-pyridylacetyl, 4-pyridylacetyl, 2-furylacryloyl, 3-furylacryloyl, 2- thienylacryloyl, 3-thienylacryloyl, piperidinoacetyl and 4-methylpiperidinoacetyl, 2-amino-3-(indol-2- yl)propionyl and 2-amino-3-(indol-3-yl)propionyl groups.

Where the acyl group represented by R3, R4 or R5 is a sulphonyl group, this may be an aliphatic sulphonyl group (for example a methanesulphonyl or ethanesulphonyi group) or an aromatic sulphonyl group (e.g. a benzene-suiphonyl or toluenesulphonyl group).

Where the acyl group represented by R3, R4 or R5 is a phosphoryl group, this may be an aliphatic phosphoryl group (e.g. a dimethylphosphoryl or diethylphosphoryl group), an aromatic phosphoryl group (e.g. a ditolylphosphoryl group) or an araliphatic phosphoryl group (e.g. a dibenzylphosphoryl, pmethylbenzylphosphoryl, p-bromobenzylphosphoryl or p-methoxybenzylphosphoryl group).

Of the compounds of the invention, those compounds are preferred where: R' represents a hydrogen atom or a methyl group; R2 represents a hydrogen atom, a metal atom (particularly an alkali metal atom, an alkaline earth metal atom or an aluminium, zinc, iron or germanium atom), an ammonium group, an alkyl- substituted ammonium group, a group capable of forming a salt derived from a basic amino acid, an alkyl group (preferably a straight or branched chain C1-C8 alkyl group), an aralkyl group optionally having one or more substituents on the aryl moiety or a phenacyl group optionally having one or more substituents on the phenyl moiety, those substituents being selected from C1-C4 alkyl groups, C,C4 alkoxy groups, halogen atoms and trifluoromethyl groups; and R3, R4 and R5 are the same or different and each represents a hydrogen atom, an aliphatic acyl group (preferably a C22o alkanoyl group, a C3-C20 alkenoyl group or a C3-C20 alkynoyl group), an aromatic acyl group (preferably a C7-C15 group) or an araliphatic acyl group (preferably a C8 or C9 aralkanoyl group or a C9 aralkenoyl group) provided that, when R3 represents a 2-methylbutyryl group, R4 and R5 both represent acyl groups.

More preferred compounds of formula (I) are those in which: R' represents a hydrogen atom or a methyl group; R2 represents a hydrogen atom, an alkali metal atom, an alkaline earth metal atom, an ammonium group, a group capable of forming a salt derived from a basic amino acid, a C1-C4 alkyl group, a benzyl group or a phenacyl group; and H3, H4 and R5 are the same or different and each represents a hydrogen atom, a C2-C6 alkanoyl group, a C3-C6 alkenoyl group or a benzoyl group, provided that, when R3 represents a 2methylbutyryl group, both R4 and R5 represent acyl groups.

The most preferred compounds are those in which: R' represents a hydrogen atom or a methyl group (particularly a hydrogen atom); R2 represents a C1-C4 alkyl group, a benzyl group or a phenacyl group (particularly a C1-C4 alkyl group); R3 represents a C2-C6 alkanoyi group or a C3-C6 alkenoyl group (particularly a C2-C6 alkanoyl group); and R4 and R5 are the same or different and each represents a C2-C6 alkanoyl group or a benzoyl group (particularly an acetyl group or a benzoyl group).

One useful class of compounds of the present invention are those compounds of formula (II) :

in which; R' represents a hydrogen atom or a methyl group; R28 represents a hydrogen atom, an alkali metal atom, an alkaline earth metal atom, an ammonium group, a group capable of forming a salt derived from a basic amino acid, a C1-C4 alkyl group, a benzyl grou p or a phenacyl groupp R3a represents a C2-C6 alkanoyl group (other than a 2-methylbutyryl group) or a C3-C6 aikenoyl group; and m is the valence of the atom or group represented by R28.

Particularly preferred are those compounds of formula (II) in which R2a represents a C1-C4 alkyl group, a benzyl group or a phenacyl group and those compounds of formula (II) in which R2a represents an alkali metal atom.

Another particularly useful class of compounds of the present invention are those compounds of formula (III):

in which: R1 represents a hydrogen atom or methyl group; and M represents a hydrogen atom or an alkali metal atom, particularly an alkali metal atom and preferably a sodium atom.

Examples of compounds of the invention are given in the following list, in which, as explained above, the compounds are named as derivatives of ML-236A, ML-236B, MB-530A or MB-530B or thei corresponding carboxylic acids: 1. Sodium ML-236A-carboxylate 2. Calcium bis(ML-236A-carboxylate) 3. Sodium MB-530A-carboxylate 4. Calcium bis(MB-530A-carboxylate) 5. Methyl ML-236A-carboxylate 6. Ethyl ML-236A-carboxylate 7. Butyl ML-236A-carboxylate 8. Benzyl ML-236A-carboxylate 9. p-Bromobenzyl ML-236A-carboxylate 10. p-Methoxybenzyl ML-23 6A-carboxylate 11. Phenacyl ML-236A-carboxylate 12. p-Methoxyphenacyl ML-236A-carboxylate 13. Methyl MB-530A-carboxylate 14. Ethyl MB-530A-carboxylate 15. Butyl MB-53OA-carboxylate 16. Benzyl MB-530A-carboxylate 17. p-Methoxybenzyl-MB-530A-carboxylate 18.Phenacyl MB-530A-carboxylate 19. p-Methoxyphenacyl MB-530A-carboxylate 20. Sodium 8'-0-acetyl-ML-236A-carboxylate 21. Methyl 8'-O-acetyl-ML-236A-carboxylate 22. Ethyl 8'-0-acetyl-ML-236A-carboxylate 23. Butyl 8'-O-acetyl-ML-236A-carboxylate 24. Benzyl 8'-O-acetyl-M L-23 6A-carboxylate 25. p-Methoxybenzyl 8'-0-acetyl-ML-236A-carboxylate 26. Phenacyl 8'-O-acetyl-ML-236A-carboxylate 27. p-Methoxyphenacyl 8'-O-acetyl-ML-236A-carboxylate 28. Sodium 8'-0-acetyl-MB-530A-carboxylate 29. Methyl 8'-0-acetyl-MB-530A-carboxylate 30. Ethyl 8'-0-acetyl-MB-530A-carboxylate 31. Butyl 8'-O-acetyl-MB-530A-carboxylate 32. Benzyl 8'-O-acetyl-M 8-5 30A-carboxylate 33. p-Methoxybenzyl 8'-O-acetyl-MB-530A-carboxylate 34.Phenacyl 8'-O-acetyl-MB-530A-carboxylate 35. p-Methoxyphenacyl 8'-O-acetyl-MB-530A-carboxylate 36. Sodium 8'-O-propionyl-ML-236A-carboxylate 37. Methyl 8'-O-propionyl-ML-236A-carboxylate 38. Ethyl 8'-O-propionyl-ML-236A-carboxylate 39. Benzyl 8'-O-propionyl-ML-236A-carboxylate 40. Phenacyl 8'-O-propionyl-ML-236A-carboxylate 41. Sodium 8'-O-propionyl-MB-530A-carboxylate 42. Methyl 8'-O-propionyl-MB-530A-carboxylate 43. Ethyl 8'-O-propionyl-MB-530A-carboxylate 44. Benzyl 8'-O-propionyl-MB-530A-carboxylate 45. Benzyl 8'-O-propionyl-MB-530A-carboxylate 46. Phenacyl 8'-O-propionyl-MB-530A-carboxylate 47. Sodium 8'-O-butyryl-ML-236A-carboxylate 48. Magnesium bis(8'-O-butyryl-ML-236A-carboxylate) 49. Aluminium tris(8'-O-butyryl-ML-236A-carboxylate) 50. Methyl 8'-O-butyryl-ML-236A-carboxylate 51. Ethyl 8'-O-butyryl-ML-236A-carboxylate 52. Butyl 8'-O-butyryl-ML-236A-carboxylate 53.Benzyl 8'-0-butyryl-ML-236A-carboxylate 54. Phenacyl 8'-O-butyryl-ML-236A-carboxylate 55. Dimethylaminoethyi 8'-0-butyryl-ML-236A-carboxylate 56. Sodium 8'-0-butyryl-MB-530A-carboxylate 57. Calcium bis(8'-O-butyryl-MB-530A-carboxylate) 58. Methyl 8'-O-butyryl-MB-530A-carboxylate 59. Ethyl 8'-0-butyryl-MB-530A-carboxylate 60. Butyl 8'-O-butyryl-M 8-530A-carboxylate 61. Benzyl 8'-O-butyryl-MB-530A-carboxylate 62. Phenacyl "-O-butyryl-M B-530A-ca rboxylate 63. Sodium 8'-O-(4-pentenoyl)-ML-236A-carboxylate 64. Methyl 8'-O-(4-pentenoyl)-ML-236A-carboxylate 65. Ethyl 8'-O-(4-pentenoyl)-ML-236A-carboxylate 66. Butyl 8'-O-(4-pentenoyl)-ML-236A-carboxylate 67. Benzyl 8'-O-(4-pentenoyl)-ML-236A-carboxylate 68.Phenacyl 8'-O-(4-pentenoyl)-ML-23 6A-carboxylate 69. Sodium 8'-O-(4-pentenoyl)-MB-530A-carboxylate 70. Methyl 8'-O-(4-pentenoyl)-MB-530A-carboxylate 71. Ethyl 8'-0-(4-pentenoyl)-MB-530A-carboxylate 72. Butyl 8'-O-(4-pentenoyl)-MB-530A-carboxylate 73. Benzyl 8'-O-(4-pentenoyl)-M B-530A-carboxylate 74. Phenacyl 8'-O-(4-pentenoyl)-MB-530A-carboxylate 75. Sodium 8'-O-isovaleryl-ML-236A-carboxylate 76. Methyl 8'-O-isovaleryl-ML-236A-carboxylate 77. Ethyl 8'-O-isovalery-ML-236A-carboxylate 78. Butyl 8'-O-isovalery-ML-236A-carboxylate 79. Benayl 8'-O-isovalery-ML-236A-carboxylate 80. Phenacyl 8'-O-isovaleryl-ML-236A-carboxylate 81. Sodium 8'-O-isovaleryl-MB-530A-carboxylate 82. Methyl 8'-O-isovalery-MB-530A-carboxylate 83.Ethyl 8'-O-isova leryl-M B-53 0A-ca rboxylate 84. Butyl 8'-O-isovaleryl-MB-530A-carboxylate 85. Benzyl 8'-O-isovaleryl-MB-530A-carboxylate 86. Phenacyl 8'-O-isovaleryl-MB-530A-carboxylate 87. Sodium 8'-O-hexanoyl-ML-236A-carboxylate 88. Methyl 8'-O-hexanoyl-ML-236A-carboxylate 89. Ethyl 8'-O-hexanoyl-ML-236A-carboxylate 90. Butyl 8'-O-hexanoyl-ML-236A-carboxylate 91. Benzyl 8'-O-hexanoyl-ML-236A-carboxylate 92. Phenacyl 8'-O-hexanoyl-ML-236A-carboxylate 93. Sodium 8'-O-hexanoyl-MB-530A-carboxylate 94. Methyl 8'-O-hexanoyl-MB-530A-carboxylate 95. Ethyl 8'-O-hexa noyl-MB-53 0A-carboxylate 96. Butyl 8'-O-hexanoyl-MB-530A-carboxylate 97. Benzyl 8'-O-hexanoyl-MB-530A-carboxylate 98.Phenacyl 8'-Q-hexanoyl-MB-530A-carboxylate 99. Sodium 8'-O-palmitoyl-ML-236A-carboxylate 100. Methyl 8'-O-palmitoyl-ML-236A-carboxylate 101. Benzyl 8'-O-palmitoyl-ML-236A-carboxylate 102. Phenacyl 8'-O-palmitoyl-ML-236A-carboxylate 103. Sodium 8'-O-palmitoyl-M B-53OA-carboxylate 104. Methyl 8'-O-palmitoyl-MB-530A-carboxylate 105. Ethyl 8'-O-pa I m itoyl-M B-53 0A-carboxylate 106. Benzyl 8'-O-palmitoyl-MB-530A-carboxylate 107. Sodium 8'-O-stearoyl-ML-236A-carboxylate 108. Sodium 8'-O-stearoyl-MB-530A-carboxylate 109. Sodium 8'-O-linolenoyl-NiL-236A-carboxylate 110. Methyl 8'-O-linolenoyl-ML-236A-carboxylate 111. Sodium 8'-O-linolenoyl-MB-530A-carboxylate 112.Methyl 8'-O-linolenoyl-MB-530A-carboxylate 113. Sodium 8'-O-benzoyl-ML-236A-carboxylate 114. Potassium 8'-O-benzoyl-ML-236A-carboxylate 11 5. Aluminium tris(8'-O-benzoyl-ML-236A-carboxylate) 116. Methyl 8'-O-benzoyl-ML-236A-carboxylate 117. Ethyl 8'-O-benzoyl-ML-236A-carboxylate 118. Bulyl 8'-O-benzoyl-ML-236A-carboxylate 119. Benzyl 8'-O-benzoyl-ML-236A-carboxylate 120. p-Methoxybenzyl 8'-O-benzoyl-M L-236A-carboxylate 121. Phenacyl 8'-O-benzoyl-ML-236A-carboxylate 122. p-Methoxyphenacyl 8'-O-benzoyl-ML-236A-carboxylate 123. Sodium 8'-O-benzoyl-M B-530A-carboxylate 124. Potassium 8'-O-benzoyl-MB-530A-carboxylate 125. Aluminium tris(8'-O-benzoyl-MB-530A-carboxylate) 126. Methyl 8'-O-benzoyl-MB-530A-carboxylate 127. Ethyl 8'-O-benzoyl-MB-530A-carboxylate 128. Butyl 8'-O-benzoyl-MB-530A-carboxylate 129. Benzyl 8'-O-benzoyl-MB-530A-carboxylate 130. p-Methoxybenzyl 8'-O-benzoyl-M B-530A-carboxylate 131. Phenacyl 8'-O-benzoyl-M B-530A-carboxylate 132. p-Methoxyphenacyl 8'-O-benzoyl-M B-530A-carboxylate 133. Sodium 8'-O-(p-toluoyl)-ML-23 6A-carboxylate 134. Calcium bis[8'-O-(p-toluoyl)-ML-236A-carboxylate] 135. Ethyl 8'-O-(p-toluoyl)-M L-23 6A-ca rboxylate 136. Butyl 8'-O-(p-toluoyi)-ML-236A-carboxylate 137. Benzyl 8'-O-(p-toluoyl)-ML-236A-carboxylate 138. Phenacyl 8'-O-(p-toluoyl)-ML-236A-carboxylate 139.Sodium 8'-O-(p-toluoyl)-MB-530A-carboxylate 140. Methyl 8'-O-(p-toluoyl)-MB-530A-carboxylate 141. Butyl 8'-O-(p-toluoyl)-M B-530A-carboxylate 142. Benzyl 8'-O-(p-toluoyl)-M B-530A-carboxylate 143. Phenacyl 8'-O-(p-toluoyl)-MB-530A-carboxylate 144. Sodium 8'-O-(2-chlorobenzoyl)-ML-236A-carboxylate 145. Sodium 8'-O-(2-chlorobenzoyl)-MB-530A-carboxylate 146. Sodium 8'-O-(3-chlorobenzoyl)-ML-236A-carboxylate 147. Sodium 8'-O-(3-chlorobenzoyl)-MB-530A-carboxylate 148. Sodium 8'-O-(4-chlorobenzoyl)-ML-236A-carboxylate 149. Sodium 8'-O-(4-chlorobenzoyl)-MB-530A-carboxylate 150. Sodium 8'-O-salicyloyl-ML-236A-carboxylate 151. Potassium 8'-O-salicyloyl-ML-236A-carboxylate 152. Methyl 8'-O-salicyloyl-ML-236A-carboxylate 153.Ethyl 8'-O-salicyloyl-ML-236A-carboxylate 154. Butyl 8'-O-salicyloyl-ML-236A-carboxylate 155. Benzyl 8'-O-salicyloyl-M L-23 6A-ca rboxylate 156. Phenacyl 8'-O-salicyloyl-ML-236A-carboxylate 157. Sodium 8'-O-salicyloyl-MB-236A-carboxylate 158. Potassium 8'-O-salicyloyl-M B-530A-carboxylate 159. Methyl 8'-0-salicyloyl-MB-530A-carboxylate 160. Ethyl 8'-O-salicyloyl-MB-236A-carboxylate 161. Butyl 8'-O-salicyloyl-MB-530A-carboxylate 162. Benzyl 8'-O-salicyloyl-MB-530A-carboxylate 163. Phenacyl 8'-0-salicyloyl-MB-530A-carboxylate 1 64. Sodium 8'-O-phenylacetyl-ML-236A-carboxylate 165.Methyl 8'-O-phenylacetyl-ML-236A-carboxylate 1 66. Ethyl 8'-0-phenylacetyl-ML-236A-carboxylate 167. Benzyl 8'-O-phenylacetyl-ML-23 6A-carboxylate 168. Sodium 8'-0-phenylacetyl-MB-530A-carboxylate 169. Methyl 8'-O-phenylacetyl-M B-530A-carboxylate 170. Ethyl 8'-O-(-chlorobenzoyl)-MB-236A-carboxylate 171. Benzyi 8'-0-phenylacetyl-MB-530A-carboxylate 172. Sodium 8'-O-(-chlorobenzoyl)-ML-236A-carboxylate 173. Sodium 8'-O-cinnamoyl-MB-53OA-carboxylate 174. Sodium 8'-O-(p-hydroxycinnamoyl)-ML-236A-carboxylate 175. Sodium 8'-O-(p-hydroxyclnnamoyl)-MB-530A-carboxylate 176. Sodium 8'-O-cyclohexanecarbonyl-ML-236A-carboxylate 177.Sodium 8'-O-cyclohexanecarbonyl-MB-530A-carboxylate 178. Sodium 8'-0-(2-thenoyl)-ML-236A-carboxylate 179. Methyl 8'-O-(2-thenoyl)-ML-236A-carboxylate 180. Ethyl 8'-O-(2-thenoyl)-ML-236A-carboxylate 181. Sodium 8'-O-(2-thenoyl)-MB-530A-carboxylate 182. Methyl 8'-O-(2-thenoyl)-MB-530A-carboxylate 183. Ethyl 8'-O-(2-thenoyl)-MB-530A-carboxylate 184. Sodium 8'-O-(2-furoyl)-ML-236A-carboxylate 185. Methyl 8'-O-(2-furoyl)-ML-236A-carboxylate 186. Ethyl 8'-O-(2-furoyl)-ML-236A-carboxylate 187. Sodium 8'-O-(2-furoyl)-MB-530A-carboxylate .188. Methyl 8'-0-(2-furoyl)-MB-530A-carboxylate 189. Ethyl 8'-O-(2-furoyl)-MB-530A-carboxylate 1 90. Sodium 8'-O-(2-thienylacetyl)-ML-236A-carboxylate 191. Methyl 8'-O-(2-thienylacetyl)-ML-236A-carboxylate 192. Ethyl 8'-O-(2-thienylacetyl)-ML-236A-carboxylate 193.Butyl 8'-O-(2-thienylacetyl)-ML-236A-carboxylate 194. Benzyl 8'-O-(2-thienylacetyl)-ML-236A-carboxylate 195. Phenacyl 8'-O-(2-thienylacetyl)-ML-236A-carboxylate 196. Sodium 8'-O-(2-thienylacetyl)-MB-530A-carboxylate 197. Methyl 8'-O-(2-thienylacetyl)-MB-530A-carboxylate 198. Ethyl 8'-O-(2-thienylacetyl)-MB-530A-carboxylate 199. Sodium 8'-O-methanesulphonyl-ML-236A-carboxylate 200. Methyl 8'-O-methanesulphonyl-ML-236A-carboxylate 201. Ethyl 8'-O-methanesulphonyl-ML-236A-carboxylate 202. Sodium 8'-O-methanesulphonyl-MB-530A-carboxylate 203. Methyl 8'-O-methanesulphonyl-MB-530A-carboxylate 204. Ethyl 8'-O-methanesulphonyl-MB-530A-carboxylate 205. Trisodium 8'-O-phosphoryl-ML-236A-carboxylate 206. Trisodium 8'-O-phosphoryl-MB-530A-carboxylate 207. Sodium 3-0-acetyl-ML-236A-carboxylate 208.Methyl 3-0-acetyl-ML-236A-carboxylate 209. Ethyl 3-0-acetyl-ML-236A-carboxylate 210. Butyl 3-O-acetyl-ML-236A-carboxylate 211. Benzyl 3-O-acetyl-ML-236A-carboxylate 212. Sodium 3-0-acetyl-MB-530A-carboxylate 213. Methyl 3-O-acetyl-MB-530A-carboxylate 214. Butyl 3-O-acetyl-MB-530A-carboxylate 215. Benzyl 3-O-acetyl-MB-530A-carboxylate 21 6. Sodium 3-O-butyryl-ML-236A-carboxylate 217. Methyl 3-0-butyryl-ML-236A-carboxylate 218. Butyl 3-O-butyryl-ML-236A-carboxylate 219. Benzyl 3-O-butyryl-ML-236A-carboxylate 220. Sodium 3-0-butyryl-MB-530A-carboxylate 221. Methyl 3-O-butyryl-M B-530A-ca rboxyla'te 222.Ethyl 3-O-butyryl-MB-530A-carboxylate 223. Benzyl 3-0-butyryl-MB-530A-carboxylate 224. Sodium 3-O-benzoyl-ML-236A-carboxylate 225. Methyl 3-O-benzoyl-M L-2 3 6A-carboxylate 226. Ethyl 3-O-benzoyl-M L-23 6A-carboxylate 227. Butyl 3-O-benzoyl-ML-236A-carboxylate 228. Benzyl 3-0-benzoyl-ML-236A-carboxylate 229. Sodium 3-0-benzoyl-N1B-530A-carboxylate 230. Methyl 3-O-benzoyl-M B-53 0A-carboxylate 231. Ethyl 3-O-benzoyl-MB-530A-carboxylate 232. Butyl 3-O-benzoyl-M B-530A-ca rboxylate 233. Benzyl 3-O-benzoyl-M B-530A-carboxylate 234. Sodium 3,8'-di(O-acetyl)-ML-236A-carboxylate 235. Methyl 3,8'-di-(O-acetyl)-ML-236A-carboxylate 236.Ethyl 3,8'-di(O-acetyl)-ML-236A-carboxylate 237. Benzyl 3,8'-di-(O-acetyl)-ML-236A-carboxylate 238. Sodium 3,8'-di(O-acetyl)-MB-530A-carboxylate 239. Methyl 3,8'-di-(O-acetyl)-MB-530A-carboxylate 240. Butyl 3,8'-di(O-acetyl)-MB-530A-carboxylate 241. Benzyl 3,8'-di(O-acetyl)-MB-530A-carboxylate 242. Sodium 3,8'-di-(O-butyl)-ML-236A-carboxylate 243. Methyl 3;8'-di(O-butyryl)-ML-236A-carboxylate 244. Butyl 3,8'-di(O-butyl)-ML-236A-carboxylate 245. Benzyl 3,8'-di(O-butyryl)-ML-236A-carboxylate 246. Sodium 3,8'-di(O-butyryl)-MB-530A-carboxylate 247. Methyl 3,8'-di(O-butyryl)-MB-530A-carboxylate 248. Butyl 3,8'-di(O-butyryl)-MB-530A-carboxylate 249. Benzyl 3,8'-di(O-butyryl)-MB-530A-carboxylate 250. Sodium 3,8'-di(O-benzoyl)-ML-236A-carboxylate 251. Methyl 3,8'-di(O-benzoyl)-ML-236A-carboxylate 252. Ethyl 3,8'-di-(0-benzoyl)-ML-236A-carboxylate 253. Butyl 3,8'-di(O-benzoyl-8'-O-butyryl-ML-236A-carboxylate 254. Benzyl 3,8'-di(O-benzoyl-8'-O-butyryl-ML-236A-carboxylate 255. Sodium 3,8'-di(O-benzoyl)-MB-530A-carboxylate 256. Methyl 3,8'-di(O-benzoyl)-MB-530A-carboxylate 257. Butyl 3,8'-di(O-benzoyl)-MB-530A-carboxylate 258. Benzyl 3,8'-di(O-benzoyl)-MB-530A-carboxylate 259. Sodium 3-O-acetyl-8'-O-butyryl-ML-236A-carboxylate 260. Sodium 3-O-acetyl-8'-O-butyryl-MB-236A-carboxylate 261. Sodium 3-O-butyryl-8'-O-acetyl-ML-236A-carboxylate 262. Sodium 3-O-butyryl-8'-0-acetyl-M B-53 0A-carboxylate 263. Sodium 3-0-benzoyl-8'-0-acetyl-M L-23 6A-carboxylate 264. Sodium 3-O-benzoyl-8'-O-acetyl-M B-530A-carboxylate 265. Sodium 3-O-benzoyl-8'-O-butyryl-ML-236A-carboxylate 266.Sodium 3-O-benzoyl-8'-O-butyryl-M B-530A-carboxylate 267. Sodium 3-O-acetyl-8'-O-benzoyl-M L-23 6A-carboxylate 268. Sodium 3-O-acetyl-8'-O-benzoyl-M B-530A-carboxylate 269. Sodium 3-O-butyryl-8'-O-benzoyl-M 1 6A-carboxylate 270. Sodium 3-O-butyryl-8'-O-benzoyl-MB-530A-carboxylate 271. Sodium 3,5.8'-tri(O-acetyl)-ML-236A-carboxylate 272. Potassium 3,5,8'-tri(O-acetyl)-ML-236A-carboxylate 273. Methyl 3,5,8'-tri(O-acetyl)-ML-23 6A-carboxylate 274. Ethyl 3,5,8'-tri(O-acetyl)-M L-23 6A-carboxylate 275. Butyl 3,5,8'-tri(O-acetyl )-M L-23 6A-carboxylate 276. Benzyl 3,5,8'-tri(O-acetyl)-ML-236A-carboxylate 277. Phenacyl 3,5,8'-tri(O-acetyl)-ML-236A-carboxylate 278.Sodium 3,5,8'-tri(O-butyryl)-ML-23 6A-carboxylate 279. Methyl 3,5,8'-tri(O-butyryl)-M L-236A-carboxylate 280. Ethyl 3,5,8'-tri(O-butyryl)-M L-23 6A-carboxylate 281. Butyl 3,5,8'-tri(O-butyryl)-ML-236A-carboxylate 282. Benzyl 3,5,8 '-tri(O-butyryl)-M L-236A-carboxylate 283. Pivaloyloxymethyl 3,5,8'-tri(O-butyryi)-ML-236A-carboxylat 284. Phenacyl 3,5,8'-tri(O-butyryl)-ML-236A-carboxylate 285. Sodium 3,5,8'-tri(O-butyryl)-MB-530A-carboxylate 286. Potassium 3,5,8'-tri(O-butyryl)-MB-530A-carboxylate 287. Methyl 3,5,8'-tri(O-butyryl)-MB-530A-carboxylate 288. Ethyl 3,5,8'-tri(O-butyryl)-MB-530A-carboxylate 289. Butyl 3,5.8'-tri(O-butyryl)-MB-530A-carboxylate 290.Benzyl 3,5,8'-tri(O-buityryl)-MB-530A-carboxylate 291. Pivaloyloxymethyl 3,5,8'-tri(O-butyryl)-M-530A-carboxylate 292. Phenacyl 3,5,8'-tri(O-butyryl)-MB-530A-carboxylate 293. Sodium 3,5-di(O-acetyl)-ML-236B-carboxylate 294. Potassium 3,5-di(O-acetyl)-ML-23 6B-carboxylate 295. Aluminium tris[3,5-di(O-acetyl)-ML-236B-carboxylate] 296. Calcium bis[3,5-di(O-acetyl)-ML-236 B-carboxylate] 297. Methyl 3,5-di(O-acetyl)-ML-236B-carboxylate 298. Ethyl 3,5-di(O-acetyl)-M L-23 6 B-ca rboxylate 299. Isopropyl 3,5-di(O-acetyl)-M L-23 6B-carboxylate 300. Butyl 3,5-di(O-acetyl)-ML-23 6 B-carboxylate 301. Benzyl 3,5-di(O-acetyl)-ML-236B-carboxylate 302.Pivaloyloxymethyl 3,5-di(O-acetyl)-M L-236 B-carboxylate 303. Ethoxymethyl 3,5-di(O-acetyl)-ML-236B-carboxylate 304. Phenacyl 3,5-di(O-acetyl)-ML-236B-carboxylate 305. Sodium 3,5-di(O-acetyl)-MB-530B-carboxylate 306. Potassium 3,5-di(O-acetyl)-MB-530B-carboxylate 307. Calcium bis [3,5-di(O-acetyl)-MB-530B-carboxylate 308. Aluminium tri[3,5-di(O-acetyl)-MB-530B-carboxylate 309. Methyl 3,5-di(O-acetyl)-MB-530B-carboxylate 310. Ethyl 3,5-di(O-acetyl)-MB-530B-carboxylate 311. Isopropyl 3,5-di(O-acetyl)-MB-530B-carboxylate 312. Butyl 3,5-di(O-acetyl)-MB-530B-carboxylate 313. Benzyl 3,5-di(O-acetyl)-MB-530B-acrboxylate 314. Pivaloyloxymethyl 3,5-di(O-acetyl)-MB-530B-carboxylate 315. Methoxymethyl 3,5-di(O-acetyl)-MB-530B-carboxylate 316. Ethoxymethyl 3,5-di(O-acetyl)-MB-530B-carboxylate 317.Phenacyl 3,5-di(O-acetyl)-MB-530B-carboxylate 318. Sodium 3,5-di(O-propionyl)-ML-236B-carboxylate 319. Methyl 3,5-di(O-propionyl)-ML-236B-carboxylate 320. Butyl 3,5-di(O-propionyl)-ML0236B-carboxylate 321. Benzyl 3,5-di(O-propionyl)-ML-236B-carboxylate 322. Sodium 3,5-di(O-propionyl)-MB-530B-carboxylate 323. Methyl 3,5-di(O-propionyl)-M B-530B-carboxylate 324. Butyl 3,5-di(O-propionyl)-MB-530B-carboxylate 325. Benzyl 3,5-di(O-propionyl)-MB-530B-carboxylate 326. Sodium 3,5-di(O-butyryl)-ML-236B-carboxylate 327. Methyl 3,5-di(O-butyryl)-ML-236B-carboxylate 328. Sodium 3,5-di(O-butyryl)-MB-530B-carboxylate 329. Methyl 3,5-di(O-butyryl)-MB-530B-carboxylate 330. Sodium 3,5-di(O-va leryl)-ML-23 6B-carboxylate 331. Potassium 3,5-di(O-valeryl)-ML-23 6B-carboxylate 332.Sodium 3,5-di(O-valeryl)--MB-530B-carboxylate 333. Potassium 3,5-di(O-valeryl)-MB-530B-carboxylate 334. Sodium 3,5-di(O-isova leryl)-ML-236B-carboxylate 335. Sodium 3,5-di(O-isovaleryl)-MB-530B-carboxylate 336. Sodium 3,5-di(O-stearoyl)-ML-236B-carboxylate 337. Sodium 3,5-di(O-stea royl)-M B-530B-carboxylate 338. Sodium 3,5-di(O-oleoyl)-ML-23 6B-carboxylate 339. Sodium 3,5-di(O-linoleoyl)-ML-236B-carboxylate 340. Sodium 3,5-di(O-linoleoyl)-MB-530B-carboxylate 341. Sodium 3,5-di(O-benzoyl)-ML-236B-carboxylate 342. Sodium 3,5-di(O-benzoyl)-MB-530B-carboxylate 343. Sodium 3,5-di(O-phenylacetyl)-ML-236B-carboxylate 344. Methyl 3,5-di(O-phenylacetyl)-ML-23 6B-carboxylate 345. Sodium 3,5-di(O-phenylacetyl)-MB-530B-carboxyla te 346.Methyl 3,5-di(O-phenylacetyl)-MB-530B-carboxylate 347. Sodium 3,5-di(O-cyclopentanecarbonyl)-ML-23 6B-carboxylate 348. Sodium 3,5-di(O-cyclopentanecarbonyl)-MB-530B-carboxylate 349. Sodium 3,5-di(O-cyclohexanecarbonyl)-ML-236B-carboxylate 350. Sodium 3,5-di(O-cyclohexanecarbonyl)-MB-530B-carboxylate 351. Sodium 3,5-di(O-thenoyl)-ML-236B-carboxylate 352. Sodium 3,5-di(O-thenoyl)-MB-530B-carboxylate 353. Sodium 3,5-di(O-furoyl)-ML-236B-carboxylate 354. Sodium 3,5-di(O-furoyl)-MB-530B-carboxylate 355. Sodium 3,5-di(O-thienylacetyl)-ML-236B-carboxylate 356. Sodium 3,5-di(O-thienylacetyl)-MB-530B-carboxylate 357. Sodium 3,5-di(O-methanesulphonyl)-ML-236B-carboxylate 358. Sodium 3,5-di(O-methanesulphonyl)-MB-530B-carboxylate 359.Trisodium 3,5-di(O-phosphoryl)-ML-236B-carboxylate 360. Trisodium 3,5-di(O-phosphoryl)-MB-530B-carboxylate 361. Sodium 3,5-di(O-acetyl)-8'-O-propionyl-ML-23 6A-carboxylate 362. Methyl 3,5-di(O-acetyl)-8'-0-propionyl-ML-236A-carboxylate 363. Butyl 3,5-di(O-acetyl)-8'-O-propionyl-ML-23 6A-carboxylate 364. Benzyl 3,5-di(O-acetyl)-8'-O-propionyl-ML-23 6A-carboxylate 365. Sodium 3,5-di(O-acetyl)-8'-O-propionyl-MB-530A-carboxylate 366. Methyl 3,5-di(O-acetyl)-8'-O-propionyl-tVtB-530A-carboxylate 367. Sodium 3,5-di(O-acetyl)-8'-O-butyryl-ML-236A-carboxylate 368. Methyl 3,5-di(O-acetyl)-8'-O-butyryl-M L-23 6A-carboxylate 369.Butyl 3,5-di(O-acetyl)-8'-O-butyryl-ML-236A-carboxylate 370. Sodium 3,5-di(O-acetyl)-8'-O-butyryl-MB-530A-carboxylate 371. Methyl 3,5-di(O-acetyl)-8'-O-butyryl-MB-530A-carboxylate 372. Butyl 3,5-di(O-acetyl)-8'-O-butyryl-MB-530A-carboxylate 373. Sodium 3,5-di(O-butyryl)-8'-O-acetyl-ML-236A-carboxylate 374. Sodium 3,5-di(O-butyryl)-8'-O-acetyl-MB-530A-carboxylate 375. Sodium 3,5-di(O-acetyl)-8'-O-benzoyl-ML-23 6A-carboxylate 376. Sodium 3,5-di(O-acetyl)-8'-O-benzoyl-MB-530A-carboxylate 377. Sodium 3,5-di(O-benzoyl)-8'-O-acetyl-ML-23 6A-carboxylate 378. Methyl 3,5-di(O-benzoyl)-8'-O-acetyl-ML-236A-carboxylate 379. Sodium 3,5-di(O-benzoyl)-8'-O-acetyl-MB-530A-carboxylate 380. Methyl 3,5-di(O-benzoyl )-8'-0-acetyl-M B-530A-carboxyiate Of these compounds, particularly preferred compounds are the following: Sodium ML-236A-carboxylate Sodium MB-530A-carboxylate Sodium 8'-O-butyryl-M L-23 6A-carboxylate Sodium 8'-0-hexa noyl-M L-23 6A-ca rboxylate Ethyl 8'-0-butyryl-M L-23 6A-carboxylate Ethyl 8'-0-hexanoyl-M L-23 6A-ca rboxyl ate Butyl 8'-0-butyryl-ML-236A-carboxylate Butyl 8'-O-(4-pentenoyl)-M L-23 6A-carboxylate Butyl 8'-O-isovaleryl-MB-530A-carboxylate Benzyl 8'-O-butyryl-MB-236A-carboxylate Benzyl 8'-0-hexa noyl-ML-23 6A-carboxylate Phenacyl 8'-O-butyryl-MB-530A-carboxylate Butyl 3,5-di(O-acetyl)-M L-236B-carboxylate Butyl 3,5-di(O-benzoyl)-ML-23 6B-carboxylate.

The compounds of the invention may be prepared by a variety of processes, all of which ultimately start from ML-236A, MB-236B, MB-530A, MB-530B, ML-236A carboxylic acid and MB530A carboxylic acid, all of which may be prepared by cultivating appropriate microorganisms as described in the prior art hereinbefore referred to and as specifically illustrated hereafter in the Preparations. The chemical structures of these compounds are as follows: ML-236A ML-236B

ML-530A MB-530B

ML-236A carboxylic acid MB-530A carboxylic acid

Metal salts, and especially the alkali metal salts, of ML-236A and MB-530A may be prepared by cultivating an appropriate microorganism and then recovering the desired salt from the culture broth.

Specifically, salts of ML-236A carboxylic acid may be prepared by cultivating an ML-236Aproducing microorganism of the genus Penicillium and recovering the ML-236A carboxylic acid salt from the culture broth, whilst MB-530A carboxylic acid salts may be prepared by cultivating an MB530A- producing microorganism of the genus Monascus and recovering the MB-530A carboxylic acid salt from the culture broth.

The microorganisms of the genus Penicillium are preferably Penicillium citrinum, Penicillium chrysogenum or Penicillium notatum, most preferably Penicillium citrinum SANK 1 8767 (FERM 2609), Penicillium citrinum SANK 24467, Penicillium citrinum SANK 24567, Penicillium chrysogenum SANK 12768 (ATCC 10002) orPenicillium notatum SANK 24867.

The microorganisms of the genus Monascus are preferably strains of Monascus ruber, Monascus anka, Monascus paxii, Monascus purpureus or Monascus vitreous, preferably Monascus ruber SANK 11272 (IFO 9203),MonascusruberSANK 17075 (CBS 832.70),MonascusruberSANK 17175 (CBS 503.70), Monascus ruber SANK 17275 (ATCC 18199), Monascus ruber SANK 15177 (FERM 4956), Monascus ruber SANK 13778 (FERM 4959), Monascus ruber SANK 10671 (FERM 4958), Monascus ruber SANK 18174 (FERM 4957 ), Monascus anka SANK 10171 (IFO 6540), Monascus paxii SANK 11172 (IFO 8201), Monascus purpureus SANK 10271 (IFO 4513) or Monascus vitreus SANK 10960 (NIHS 609, e-609, FERM 4960).

All of these microorganisms are available from recognized culture collections, as indicated by the following codes: 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 Hygenic Sciences, Japan; CBS =Centraal Bureau Voor Schimmelcultures, Netherlands, ATCC =American Type Culture Collection, Maryland, U.S.A.

Apart from the strains of microorganisms mentioned above, any microorganism of the genus Penicillium orMonascus, including varieties and mutants, which are capable of producing ML-236A or MB-530A, respectively, may be employed in the process of the present invention.

The ML-236A and MB-530A salts may be produced by cultivating the chosen microorganism in a culture broth under aerobic conditions, using the techniques well-known in the art for cultivation of fungi and other microorganisms. For example, the chosen strain of Peniclllium or Monascus may first be cultivated on a suitable medium and then the produced microorganism may be collected and inoculated into and cultivated on another culture medium to produce the desired ML-236A or MB530A; the culture medium used for the multiplication of the microorganism and the culture medium used for the production of the ML-236A or MB-530A may be the same or different.Any culture medium well-known in the art for the cultivation of fungi may be employed, provided that it contains, as is well-known, the necessary nutrient materials, especially an assimilable carbon source and an assimilable nitrogen source. Examples of suitable sources of assimilable carbon are glucose, maltose, dextrin, starch, lactose, sucrose and glycerine. Of these sources, glucose and glycerine are particularly preferred for the production of ML-236A and MB-530A. 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. When producing the ML-236A or MB-530A, an inorganic salt and/or a metal salt may, if necessary, be added to the culture medium. Furthermore, if necessary, a minor amount of a heavy metal may also be added.In certain cases, it is believed possible that the fungus of the genus Penicillium or Monascus may produce the desired ML-236A or MB-530A carboxylic acid salt directly, whilst in other cases the fungus will produce the ML-236A or MB-530A first and this is then converted to the desired salt in the course of a routine separation and purification procedure. Where the salts are to be produced directly by the fungus, it is important that there should be present in the culture medium or within the body of the fungus metal ions corresponding to the metal salt which it is desired to produce.

The microorganism is preferably cultivated under aerobic conditions using cultivation methods well-known in the art, for example solid culture, shaken culture or culture under aeration and agitation.

The microorganisms will grow over a wide temperature range, e.g. from 7 to 350C, but, especially when the microorganism is grown for the purpose of producing ML-236A or MB-530A or a carboxylic acid salt thereof, the more preferred cultivation temperature is within the range from 20 to 300C.

During the cultivation of the microorganism, the production of the desired ML-236A or MB-530A or carboxylic acid salt thereof may be monitored by sampling the culture medium and measuring the physiological activity of the medium by appropriate tests. Cultivation may then be continued until a substantial accumulation of active material has been achieved in the culture medium, at which time the ML-236A or MB-530A carboxylic acid salt may be isolated and recovered from the culture medium and the tissues of the microorganism by any suitable combination of isolation techniques, chosen having regard to their physical and chemical properties.For example, any or all of the following isolation techniques may be employed: extraction of the liquor from the culture broth with a hydrophilic solvent (such as 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 off all or part of the solvent under reduced pressure; dissolution into a more polar solvent (such as an 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" (a registered Trade Mark for a material available from Pharmacia Co.Limited, U.S.A.); absorptive chromatography with active carbon or silica gel; rapid liquid chromatography; conversion to ML-236A or MB-530A itself or its parent acid; direct purification in the form of the metal salt; and other similar methods. By using a suitable combination of these techniques, the desired salt can be isolated from the culture broth as a pure substance.

As described in the prior art, ML-236A, ML-236B, MB-530A, MB-530B, ML-236A carboxylic acid and MB-530A carboxylic acid, all of which are important starting materials for the preparation of certain of the compounds of the present invention, may also be produced using the microorganisms and techniques described above.

Others of the compounds of the invention may be prepared by the following methods.

Method 1 Preparation of Salts Carboxylic acid salts of formula (IV):

(in which R1, R3 and R4 are as defined above; M1 represents the cationic moiety of a salt; and m' represents the valency of M1) may be prepared by hydrolyzing a lactone of formula (V):

(in which R1, R3 and R4 are as defined above) and, if necessary converting the carboxyl group of the product thus obtained to the corresponding salt.

The hydrolysis may be carried out by any of the methods well-known in the art for conversion of lactones to the corresponding hydroxyacid. If desired, the hydroxyacid may then be extracted with a suitable organic solvent and then salified by reaction with a base corresponding to the salt which it is desired to produce, for example with a metal hydroxide or carbonate, ammonia, organic amine or amino acid. However, where it is desired to prepare a metal salt (i.e. M' represents a metal atom), it is often most convenient to effect the hydrolysis and salification in a single step by using a basic compound of the metal, preferably the metal hydroxide, in an amount at least equimolar with respect to the lactone of formula (V), to effect hydrolysis.In this case, the metal salt of formula (IV) is produced directly and may be obtained simply by distilling off the solvent from the reaction mixture.

The hydrolysis may be effected by conventional methods, for example by contacting the lactone (V) with a dilute (e.g. from 0.1 to 02N) aqueous solution of the metal hydroxide, preferably an alkali metal hydroxide such as sodium hydroxide or potassium hydroxide, in water or an aqueous organic solution, e.g. an aqueous alcohol or aqueous dioxan. The hydrolysis may be effected over a wide range of temperatures, for example at room temperature or at an elevated temperature, e.g. from 50 to 100 C. The time required for the reaction will vary over a wide range, depending upon the reagents and the reaction temperature, but the reaction will normally require several hours.

In a preferred embodiment of this Method an alkali metal salt of formula (VI):

(in which R1 is as defined above and M2 represents an alkali metal atom) may be prepared by hydrolyzing, in an alkaline solution containing said alkali metal atom, a lactone of formula (all):

(in which R1 is as defined above). In this case, M2 preferably represents a sodium atom and the alkaline solution is preferably an aqueous solution of sodium hydroxide.

Method 2 Preparation of Ester from The Corresponding Lactone Carboxylic acid esters of formula (Vlil):

(wherein: R1 represents a hydrogen atom or a methyl group; R3 and R4 are the same or different and each represents a hydrogen atom or an acyl group, provided that, when R3 represents a 2-methylbutyryl group, R4 represents an acyl group; and R2b represents an alkyl group or an aralkyl group) may be prepared by reacting a lactone of formula (V):

(in which R1, R3 and R4 are as defined above) with an alcohol in the presence of an acid catalyst.

Suitable acid catalysts include: inorganic acids such as hydrochloric acid or sulphuric acid; Lewis acids, such as boron trifluoride; and acidic ion-exchange resins. The reaction may be carried out in the presence of a suitable inert organic solvent, for example benzene, diethyl ether or chloroform. However, where the alcohol, which is one of the reagents, is a liquid, we prefer to use this as the solvent.

Although the reaction will take place over a wide temperature range, it is preferably conducted with heating, for example at a temperature of from 500C to the boiling point of the solvent. After completion of the reaction, the desired product may be recovered from the reaction mixture by conventional methods. For example, when the catalyst employed is an ion-exchange resin, this resin is filtered off and then the solvent is distilled offto give the desired compound. If the catalyst is an inorganic acid or a Lewis acid, this is neutralized and then the solvent is distilled off, the residue is extracted with a suitable solvent, which is then distilled from the extract to give the desired product.

The same process may be employed for the preparation of other esters than alkyl and aralkyl esters, by using appropriate hydroxy- compounds or functional equivalents thereof; for example, compounds of formula (I) where R2 represents a bivalent aliphatic hydrocarbon group may be prepared by replacing the alcohol used in the above process by a glycol. Also, in place of the acid- catalyzed solvolysis process described above, it is possible to use any other conventional solvolysis technique to produce any compound of formula (I) in which R2 represents an ester residue capable of formation by solvolysis of a lactone.

Method 3 Preparation of Esters from The Corresponding Metal Salt Carboxylic acid esters of formula (IX):

(wherein: R1, R3, R4 and R5 are as defined above; R2C represents the alcoholic moiety of an ester; and p represents the valence of R2c) can be prepared by reacting a metal salt of formula (X):

(wherein R1, R3, R4 and R5 are as defined above; M3 represents a metal atom, preferably an alkali metal or alkaline earth metal atom; and q represents the valence of M3) with an esterifying agent, preferably an esterifying agent of formula R2C--X (in which X represents a halogen atom, for example a chlorine or bromine atom).

This reaction may be carried out using procedures conventional for alkylating carboxylic acid metal salts. For example, the metal salt of formula (X) may be contacted with the esterifying agent in the presence of an inert organic solvent, to give the desired ester of formula (IX). There is no particular limitation upon the nature of the solvent employed, provided that it has no adverse effect upon the reaction and preferred solvents include dimethylformamide, dimethyl sulphoxide, tetrahydrofuran, hexamethylphosphoryltriamide, acetone and methyl ethyl ketone. The reaction may be carried out over a wide range of temperatures, for example at room temperature or at an elevated temperature, but we normally prefer to carry out the reaction at about room temperature.The time required for the reaction will, of course, depend upon the reaction temperature and upon the reagents employed, but will normally range from 1 to 20 hours. The desired product may be recovered from the reaction mixture by conventional means, for example by diluting the reaction mixture with water, extracting the mixture with a water-immiscible solvent and distilling off the solvent from the extract.

The esterifying agent employed is preferably an alkyl halide, an aralkyl halide or a phenacyl halide and the metal salt of formula (X) is preferably an alkali metal salt (i.e. M3 represents an alkali metal atom). This alkali metal salt is preferably formed in situ by another reaction, for example that described in Method 1, prior to the reaction with the esterifying agent.

It is particularly preferred that the compounds employed should be those compounds in which: R1 represents a hydrogen atom or a methyl group; R3 represents a hydrogen atom, a C2-C6 alkanoyl group or a C3-C6 alkenoyl group; R4 and R5 are the same or different and each represents a hydrogen atom, a C2-C6 alkanoyl group or a benzoyl group, provided that, when R3 represents a 2-methylbutyryl group, R4 and R5 both represents acyl groups; and M3 represents an alkali metal atom; and that the esterifying agent should be a benzyl halide or a phenacyl halide.

Method 4 Preparation of 3-acyl, 5-acyl and 8'-acyl Derivatives A triacyl compound of formula (Xl):

(in which R1, R2 and n are as defined above and R3b, R4b and R5b are the same or different and each represents an acyl group) may be prepared by acylating a compound of formula (XII):

(in which R1, R2 and n are as defined above and R3C, R4c and R5C are the same or different and each represents a hydrogen atom or an acyl group, provided that at least one of R3C, R4C and R5C represents a hydrogen atom).

Compounds of formulae (XIII), (XIV) and (XV):

(wherein: R', R2 and n are as defined above; R4 represents a hydrogen atom or an acyl group; R3b represents an acyl group; and R4b represents an acyl group) may be prepared by acylating a compound of formula (XVI):

(wherein R1, R2, n and R3b are as defined above). By appropriate choice of conditions, it is possible to prepare either a single one of the compounds of formulae (XIII), (XIV) and (XV), a mixture of any two or a mixture of all three. The desired compound or compounds may then be isolated from the reaction mixture.

The acylation reaction itself is preferably carried out by either of the following processes: (A) Acylation Using a Reactive Derivative of An Acid In this process, the compound of formula (Xll) or (XVI) is contacted with a reactive derivative of the acid having the desired acyl group, for example an acid halide (such as an acid chloride or acid bromide), an acid anhydride or a mixed acid anhydride (for example a mixed acid anhydride of an acid with a chlorocarbonic ester or a sulphuric acid chloride). The reaction is preferably carried out in the presence of a solvent and also preferably in the presence of a base.There is no particular limitation upon the nature of the solvent to be employed, provided that it has no adverse effect upon the reaction and suitable solvents include chloroform, methylene chloride, diethyl ether, tetrahydrofuran and dioxan.

Preferred bases include such organic amines as pyridine, 4-(N,N-dimethylamino)pyridine, quinoline, triethylamine, N-methylmorpholine, N-methylpiperidine and N,N-dimethylaniline. Where the amine employed is a liquid at the reaction temperature, for example pyridine, it may also serve as the solvent.

The reaction is preferably carried out at room temperature or with cooling in order to control side reactions, but it will proceed at elevated temperatures. The reaction time will depend upon the reaction temperature and upon the nature of the reagents but it will normally be complete within a period of from 10 minutes to 10 hours. After completion of the reaction, the desired product may be recovered by conventional means, for example by diluting the reaction mixture with water, extracting the mixture with a water-immiscible solvent and distilling the solvent from the extract.

(B) Acylation With a Condensing Agent This reaction may be carried out under conventional conditions employing a free acid corresponding to the acyl group which it is desired to introduce in the presence of a condensing agent and, in this case, the compound of formula (XII) or (XVI) is simply contacted with the acid in the presence of the condensing agent. The condensing agent is preferably a dehydrating agent, for example a carbodiimide, such as dicyclohexylcarbodiimide. The reaction is normally carried out in the presence of a solvent, the nature of which is not critical, provided that it has no adverse effect upon the reaction. Suitable solvents include dimethylformamide, dimethylacetamide, acetonitrile, pyridine, methylene chloride, chloroform and dioxan.The reaction may be carried out over a wide temperature range, but, in order to control side reactions, we normally prefer to conduct it at room temperature or with cooling; however, the reaction will also proceed at elevated temperatures. The time required for the reaction will vary depending upon the reaction temperature and the reagents employed, but the reaction will normally be complete within a period of from 1 to 20 hours. The desired compound may then be recovered from the reaction mixture by conventional means, for example by filtering off insolubles, diluting the filtrate with water, extracting the resulting mixture with a water-immiscible solvent and then distilling the solvent from the extract to give the desired product.

In the case of both of the above acylation reactions, where the starting material contains two or more free hydroxy groups, any of the mono, di or tri- acyl derivatives can be obtained by controlling the amount of acylating agent employed. If a mixture of these compounds is obtained, the separate compounds may be isolated using conventional isolation techniques, for example chromatography on silica gel.

The compounds of the invention may ultimately be prepared from the fermentation products ML236A, ML-236B, Mb-530A and MB-530B or their corresponding carboxylic acids or carboxylic acid salts by any combination of the above reactions. These reactions are summarized in the following reaction scheme, in which the chemical structures of the compounds are simplified as follows:

In the above reaction scheme, Ac', Ac2 and Ac3 are the same or different and each represents an acyl group.

The compounds of the invention have been found to inhibit the activity of 3-hydroxy-3methylglutaryl coenzyme A reductase (HMG-CoA reductase), which is the rate- determining enzyme in the biosynthesis of cholesterol. According to the method of Knauss, et al. [J. Biol. Chem., 234, 2835 (1959)], the inhibitory activities of the compounds of the invention against the biosynthesis of cholesterol, expressed as their ID50 values (i.e. the concentration required to inhibit the biosynthesis of cholesterol by 50%), varied from 1.0 to 0.03 ,ug/ml.Moreover, sodium salts, i.e. compounds of formula (I) in which R2 represents a sodium atom, were found to be adsorbed during an in vivo test far better than the corresponding lactone and in general it is believed that the compounds of the invention have improved bioavailability compared with the corresponding lactones.

The compounds of the invention may be administered for the treatment of hypercholesterolaemia by any conventional means, but they are preferably administered in the form of tablets or capsules. The daily dose will vary depending upon the age, body weight and condition of the patient, but in general the compounds of the invention are preferably administered in an amount of from 1 mg to 10 mg per day, in a single dose or in divided doses.

The preparation of the compounds of the invention is further illustrated by the following Examples. Preparation of certain of the starting materials used in these Examples is illustrated by Preparations 1-4 and other starting materials may be prepared in a manner similar to that described in these Preparations. The preparation of ML-236A and ML-236B are described in more detail in US Patent Specification No. 3,983,140 and the preparation of MB-530 (Monacolin K) is described in United Kingdom Patent Applications No. 2046737 and No. 2049664.

Preparation 1 3-O-Butyryl-ML-236A 918 mg of ML-236A were dissolved in 5 ml of pyridine, and 1 ml of butyric anhydride was added dropwise thereto at room temperature. After leaving the reaction mixture to stand overnight, it was diluted with water and extracted with diethyl ether. The extract was washed successively with water, a saturated aqueous solution of sodium bicarbonate, water, 1 N hydrochloric acid and water and dried over anhydrous sodium sulphate. The solvent was distilled off from the solution and the residue was subjected to column chromatography through silica gel, eluted with a 5:1 by volume mixture of benzene and ethyl acetate, to give 930 mg of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C22H3205: C, 70.21%; H, 8.51%.

Found: C, 69.95%; H, 8.69%.

Nuclear Magnetic Resonance Spectrum (CDCI3) S ppm: 0.95 (3H, triplet); 4.27 (1H, multiplet); 5.32 (1 H, multiplet).

Infrared Absorption Spectrum (liquid film) Vm8x cm 3460,1740.

Preparation 2 3,8'-Di(O-butyryl)-ML-236A 306 mg of ML-236A and 0.5 ml of pyridine were dissolved in 3 ml of methylene chloride, and 0.5 ml of butyryl chloride was added dropwise thereto, with ice-cooling. The mixture was stirred at room temperature for 1 hour and then washed adding water. The organic layer was separated and dried over an hydros sodium sulphate, after which the solvent was distilled off. The resulting residue was subjected to column chromatography through silica gel, eluted with a 10:1 by volume mixture of benzene and ethyl acetate, to afford 384 mg of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C28H38O6: C, 69.96%; H, 8.52%.

Found: C, 70.14%; H, 8.31%.

Nuclear Magnetic Resonance Spectrum (CDCl3) S ppm: 0.93 (6H, triplet); 5.2-5.5 (2H, multiplet).

Infrared Absorption Spectrum (liquid film) Vmax cm 1735,1250,1175.

Preparation 3 8'-O-Butyryl-ML-236A 918 mg of ML-236A and 0.36 ml of pyridine were dissolved in 10 ml of methylene chloride. The solution was cooled in an ice bath, and 0.35 ml of butyryl chloride was added dropwise thereto. After being stirred for 1 hour, the mixture was diluted with water. The organic layer was separated, washed with water and dried over anhydrous sodium sulphate. The solvent was distilled off and the residue was subjected to column chromatography through silica gel.The 3,8'-diacyl compound (described in Preparation 2) was obtained by eluting with a 10:1 by volume mixture of benzene and ethyl acetate, the 3-acyl compound (described in Preparation 1) was obtained by eluting with a 5:1 by volume mixture of benzene and ethyl acetate, and 395 mg of the desired product were obtained as colourless crystals melting at 1 24-50C from the eluate using a 2:1 by volume mixture of benzene and ethyl acetate.

Elemental. analysis: Calculated for C22H32O3 : C, 70.2 % ; H, 8.51%.

Found: C, 70.25%; H, 8.50%.

Nuclear Magnetic Resonance Spectrum (CDCl3) S ppm: 0.95 (3H, triplet); 4.42 (1 H, multiplet).

Infrared Absorption Spectrum (Nujol-trade mark) Vmax cm-:: 3400, 1730, 1710.

Other acyl derivatives can be prepared by following the procedures described in these Preparations.

Preparation 4 Preparation of MB-530A 300 litres of a culture medium having a pH of 5.5 before sterilization and 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% ammonium chloride were charged into a 600 litre fermenter and inoculated with a culture of MonascusruberSANK 15177 (FERM 4956, NRRL 12081). Cultivation of the microorganism was continued for 120 hours at 270C with an aeration rate of 300 litres/minute and agitation at 190 revolutions per minute.

At the end of this time, the culture broth was filtered in a filter press to give a filtrate and a filter cake comprising wet cells of the microorganism.

The filtrate was adjusted to a pH of 3.0 by the addition of 6N hydrochloric acid and then extracted with 400 litres of ethyl acetate. The extract (about 400 litres) was concentrated by evaporation under reduced pressure and then dehydrated over anhydrous sodium sulphate, after which it was evaporated to dryness, to give about 60 g of an oily product. This oily product was washed with ethylcyclohexane and with hexane and the residue (20 g) was separated by chromatography using a liquid chromatography device for large volume sampling (System 500 liquid chromatography, produced by Waters Co., U.S.A.), eluted with 60% v/v aqueous methanol. Fractions having a chromatographic retention time of 6 minutes were collected and concentrated by evaporation under reduced pressure to give 100 mg of the desired MB-530A as an oily product.This oily MB-530A was recrystallized from a mixture of acetone and diethyl ether to give 57 mg of the desired product in the form of colourless needles having the following properties: 1. Melting Point: 92-930C.

2. Elemental Analysis: Calculated for C19H28O4 : C, 69.76% ; H, 8.68%.

Found: C,71.22%; H, 8.81%.

3. Molecular weight: 320 (by mass analysis).

4. Molecular formula: C19H2804.

5. Ultraviolet Absorption Spectrum: As shown in Figure 1 of the accompanying drawings.

6. Infrared Absorption Spectrum: As shown in Figure 2 of the accompanying drawings.

7. Nuclear Magnetic Resonance Spectrum: As shown in Figure 3 of the accompanying drawings.

8. Solubility: readily soluble in methanol, ethanol, acetone and ethyl acetate; soluble in benzene; insoluble in hexane and petroleum ether.

9. Colouration reaction: a pink colour is seen when a thin layer chromatogram on silica gel of the compound is developed with 50% v/v sulphuric acid.

10. Inhibitory activity against the biosynthesis of cholesterol: a 50% inhibition of the synthesis of cholesterol in a rat liver is observed at a concentration of 0.04 Mg/ml.

Example 1 Sodium ML-236A-carboxylate 0.4 g of ML-236A was added to 12 ml of 0.1 N aqueous solution of sodium hydroxide, and the mixture was heated at 80-900C for 2 hours. After. completion of the reaction, insolubles were filtered off and the filtrate was lyophilized to afford 0.41 g of the desired product as a colourless powder.

Elemental analysis: Calculated for C18H27O5Na: C, 62.43%; H, 7.80%.

Found ; C, 62.58% ; H, 7.72%.

Example 2 Sodium 8'-O-butyryl-M L-236A-carboxylate 0.75 g of 8'-0-butyryl-ML-236A was added to 20 ml of a 0.1 N aqueous solution of sodium hydroxide, and the resulting mixture was heated at 80-900C for 2 hours. After completion of the reaction, insolubles were filtered off and the filtrate was lyophilized to give 0.83 g of the desired product as a colourless powder.

Elemental analysis: Calculated for C22H330GNa: C, 63.46%; H, 7.93%; Na, 5.53%.

Found: C,63.54%, H, 7.90%; Na, 5.50%.

Nuclear Magnetic Resonance Spectrum (D20) a ppm: 0.95 (3H, triplet); 4.15 (2H, triplet); 3.5-4.5 (2H, multiplet).

Infrared Absorption Spectrum (Nujol) Pmax cm~1: 3450, 1730, 1580.

Example 3 Sodium 8'-O-hexanoyl-ML-236A-carboxylate 0.88 g of 8'-0-hexanoyl-ML-236A was added to 20 ml of a 0.1 N aqueous solution of sodium hydroxide. The mixture was treated in the same manner as described in Example 1, to give 0.84 g of the desired product as a colourless powder.

Elemental analysis: Calculated for C24H37O6Na : C, 64.86 % ; H, 8.33 % ; Na, 5.18%.

Found : C, 64.75 % ; H, 8.37 % ; Na, 5.25%.

Nuclear Magnetic Resonance Spectrum (D20) 3 ppm: 0.87 (3H, triplet); 4.13 (2H, multiplet); 3.5-4.5 (2H, multiplet).

Infrared Absorption Spectrum (Nujol) vmax cm-': 3450, 1 730, 1 580.

Example 4 Ethyl 8'-0-butyryl-ML-236A-carboxylate 0.79 g of 8'-0-butyryl-ML-236A were dissolved in 10 ml of ethanol, and 1 g of a sulphuric acidtype strongly acidic cation exchange resin (sold under the trade name Dowex 50W) was added thereto. The mixture was heated at 60-700C for 3 hours. After completion of the reaction, the mixture was filtered and the filtrate was concentrated by evaporation under reduced pressure. The residue was purified by column chromatography through silica gel to afford 0.41 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C24H38O6 : C, 68.25 % ; H, 9.00 %.

Found : C, 68.19 % ; H, 9.06 %.

Nuclear Magnetic Resonance Spectrum (CDCl3) a ppm: 0.95 (3H, triplet); 1.23 (3H, tripiet); 4.12 (2H, quartet); 3.5-4.5 (2H, multiplet).

Infrared Absorption Spectrum (liquid film) Vmax cm-.

3450,1740.

Example 5 Ethyl 8'-0-hexa noyl-ML-236A-carboxylate 0.9 g of 8'-O-hexanoyl-ML-236A was dissolved in 10 ml of ethanol, and 1.5 g of a sulphonic acid-type strongly acidic cation exchange resin were added thereto. The mixture was treated in the same manner as described in Example 3, to give 0.55 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C26H46O6 : C, 69.33 % ; H, 9.33 %.

Found : C, 68.21 % ; H, 9.39 %.

Nuclear Magnetic Resonance Spectrum (CDCl3) # ppm : 0.87 (3H, triplet); 1.25 (3H, triplet); 4.13 (2H, quartet); 3.5-4.5 (2H, multiplet).

Infrared Absorption Spectrum (liquid film) vmax cm~1: 3400,1730,1720.

Example 6 Butyl 8'-0-butyryl-ML-236A-ca rboxylate 0.79 g of 8'-0-butyryl-ML-236A was dissolved in 10 ml of butanol, and 1.0 g of a sulphonic acid-type strongly acidic cation exchange resin was added thereto. The mixture was treated in the same manner as described in Example 3, to afford 0.59 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C26H42O6: C,69.33%; H, 9.33%.

Found : C, 69.25 % ; H, 9.40 %.

Nuclear Magnetic Resonance Spectrum (CDCl3) a ppm: 0.95 (3H, triplet); 1.24 (3H, triplet); 4.12 (2H, triplet); 3.5-4.5 (2H, multiplet).

Infrared Absorption Spectrum (liquid film) vmax cm 3440, 1730.

Example 7 Butyl 8'-O-(4-pentenoyl)-ML-236A-carboxylate Following the procedure described in Example 3, but using 0.92 g of 8'-O-(4-pentenoyl)-ML 236A, 10 ml of butanol and 1.0 g of the ion exchange resin, there was obtained 0.45 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C27H42O6 : C, 70.13 % ; H, 9.09 %.

Found : C, 70.04 % ; H, 9.13 %.

Nulcear Magnetic Resonance Spectrum (CDCl3) # ppm : 1.24 (3H, multiplet); 3.5-4.5 (4H, multiplet); 4.8-6.2 (8H, multiplet); Infrared Absorption Spectrum (liquid film) vmax cm 3400,1730.

Example 8 Butyl 8'-O-isovaleryl-MB-530A-carboxylate Following the procedure described in Example 3, but using 0.9 g of 8'-O-isovaleryl-MB-530A, 10 ml of butanol and 1.0 g of the ion exchange resin, there was obtained 0.52 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C27H44O6 : C, 69.83 % ; H, 9.49 %.

Found : C, 69.77 % ; H, 9.46 %.

Nuclear Magnetic Resonance Spectrum (CDC13) S ppm: 0.89 (1 H, doublet); 1.24 (3H, triplet); 4.12 (2H, triplet); 3.5-4.5 (2H, multiplet).

Infrared Absorption Spectrum (liquid film) Vmax cm 3430,1730.

Example 9 Benzyl 8'-O-butyryl-ML-236A-carboxylate 1.0 g of sodium 8'-0-butyryl-ML-236A-carboxylate was dissolved in 5 ml of dimethylformamide, and 1 ml of benzyl chloride was added thereto. The mixture was left to stand overnight. The reaction mixture was then diluted with water and extracted with ethyl acetate. The organic layer was separated, washed with water and dried over anhydrous sodium sulphate. The solvent was distilled off under reduced pressure and the residue was purified by column chromatography through silica gel, to afford 1.2 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C28H40O6: C,71.90%; H, 8.26%.

Found : C, 71.84 %. H, 8.25 %.

Nuclear Magnetic Resonance Spectrum (CDCl3) S ppm: 0.94 (3H, triplet); 1.24 (3H, triplet); 4.11 (2H, triplet); 3.5-4.5 (2H, multiplet); 5.20 (2H, singlet); 7.42 (5H, singlet).

Infrared Absorption Spectrum (liquid film) Vmax cm 3540,1730..

Example 10 Benzyl 8'-O-hexanoyl-M L-236A-carboxylate Following the procedure described in Example 8, but using 1.2 g of sodium 8'-O-hexanoyl-ML236A-carboxylate and 1 ml of benzyl chloride, there were obtained 1.3 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C31H44O6 : C, 72.66 % : H, 8.59 %.

Found : C, 72.58 % : H, 8.50 %.

Nuclear Magnetic Resonance Spectrum (CDCl3) a ppm: 0.89 (3H, triplet); 4.14 (2H, triplet); 3.5-4.5 (2H, multiplet); 5.21 (2H, singlet); 7.44 (5H, singlet).

Infrared Absorption Spectrum (liquid film) may cm 3400,1730.

Example 11 Phenacyl 8'-0-butyryl-MB-530A-carboxylate Following the procedure described in Example 8, but using 0.86 g of sodium 8'-O-butyryl-MB530A-carboxylate and 0.45 g of phenacyl bromide, there was obtained 1.0 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C31H42O7 : C, 70.72 % ; H, 7.98 %.

Found : C, 70.65 % ; H, 8.04 %.

Nuclear Magnetic Resonance Spectrum (CDGI3) S ppm: 0.95 (3H, triplet); 4.13 (2H, triplet); 3.5-4.5 (2H, multiplet); 5.44 (2H, singlet); 7.4-8.2 (5H, multiplet).

Infrared Absorption Spectrum (liquid film) PmaX cm~1: 3450,1730,1700.

Example 12 Butyl 3,5-di(O-acetyl)-ML-236B-carboxylate 0.32 g of butyl ML-236B-carboxylate was dissolved in 1 ml of pyridine. After the addition of 1 ml of acetic anhydride, the mixture was left to stand at room temperature for 1 hour. The reaction mixture was then diluted with water and extracted with ethyl acetate. The solvent was distilled under reduced pressure from the extract and the residue was purified by column chromatography through silica gel, to afford 0.35 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C31H4808 C, 67.88 ,ó; H, 8.76%.

Found : C, 67.74 % ; H, 8.81 %.

Nuclear Magnetic Resonance Spectrum (CDCI3) S ppm: 2.00 (3H, singlet); 2.03 (3H, singlet).

Infrared Absorption Spectrum (liquid film) Vmax cm 1740.

Example 13 Butyl 3,5-di(O-benzoyl-ML-236B-carboxylate Following the procedure described in Example 11, but using 0.34 g of butyl ML-236Bcarboxylate, 0.3 ml of benzoyl chloride and 1 ml of pyridine, there was obtained 0.38 g of the desired product as a colourless oily substance.

Elemental analysis: Calculated for C41Hs2 8 C, 73.21%; H, 7.74 %.

Found : C, 73.10 % ; H, 7.89 %.

Nuclear Magnetic Resonance Spectrum (CDCI3) S ppm: 7.2-7.6 (3H, multiplet); 7.8-8.2 (2H, multiplet).

Infrared Absorption Spectrum (liquid film) vmax cm 1720.

Example 14 Sodium ML-236A-carboxylate 300 litres of a culture medium (pH 5.5 before sterilization) containing 2% w/v glucose,0.1 w/v peptone ("Kyokuto" brand) and 3% w/v malt extract were charged into a 600 litre fermenter and inoculated with the organism Penicillium citrinum SANK 1 8767. Cultivation was continued at 260C for 96 hours under aerobic conditions at an aeration rate of 300 litres per minute and agitation at 145 rotations per minute. The culture broth (containing the organism) was adjusted to pH 3.4 with 6N hydrochloric acid and extracted with 800 litres of ethyl acetate. The extract was washed with 200 litres of a saturated aqueous solution of sodium chloride and then concentrated by evaporation in a vacuum to 18 litres.The resulting solution was extracted with 600 litres of ethyl acetate. 50 g of trifluoroacetic acid were added to the extract and reacted at 800C for 30 minutes. The reaction mixture was washed successively with 20 litres of a 2% w/v aqueous solution of sodium bicarbonate and 2C litres of a 10% w/v aqueous solution of sodium chloride, after which it was concentrated by evaporation under reduced pressure, to give 130 g of an oily substance. This oil was dissolved in 400 ml of methanol. 20 ml of the methanolic solution (containing 6.5 9 of the oil) were subjected to preparative rapid liquid chromatography, System 500 type (produced by Waters Co., Ltd.) equipped with a Prepac C18 column (reversed phase column). A mixture of methanol and water (60:40 v/v) was used as the developer.The development was carried out at a flow rate of 200 ml/minute (developing time: about 10 minutes), watching a differential refractometer connected to the apparatus, the portion showing a main peak on the differential refractometer was separated. This operation was repeated and resulting main peak fractions were collected and concentrated to give 9.2 g of an oily substance, which was dissolved in 30 ml of methanol. 6 ml of the methanolic solution (containing about 1.8 g of the oil) were subjected again to the same chromatography but developed with a 55:45 v/v mixture of methanol and water at a flow rate of 200 ml/minute. A portion showing a main peak was separated.

This operation was repeated and the main peak fractions were collected and concentrated to afford 1020 mg of ML-236A as an oily substance. To this substance were added 33.2 ml of 0.1 N aqueous solution of sodium hydroxide and the mixture was stirred at 50-600C for 3 hours. Insolubles were filtered off and the filtrate was Iyophilized, to give 1090 mg of sodium ML-236A-carboxylate.

Example 15 Sodium ML-236A-carboxylate 1030 litres of an ethyl acetate extract from the culture broth obtained by the same procedures as described in Example 14 were washed with 200 litres of a saturated aqueous solution of sodium chloride. The extract was then dried over an hydros sodium sulphate and concentrated to dryness to give 120 g of an oily substance containing ML-236A carboxylic acid.

To this oil was added methanol to give a total volume of 200 ml. 20 ml of the solution was subjected to preparative rapid liquid chromatography equipped with a reversed phase column (the same as used in Example 14) and eluted with a 20% v/v aqueous methanol solution (containing 2% acetic acid) at a flow rate of 200 ml/minute. A portion showing a main peak on the differential refractometer was separated (7-10 minutes). The remaining 120 ml were treated by the same procedure. The resulting main peak fractions were collected, concentrated and extracted with ethyl acetate. The filtrate was concentrated to dryness after adding heptane, to give 8.7 g of an oily substance, which was dissolved in 20 ml of methanol and subjected to chromatography under the same conditions as above to give 930 mg of ML-236A carboxylic acid.To this product were added 2 ml of methanol and 100 ml of water and the resulting solution was adjusted to pH 8.0 with 1 N aqueous sodium hydroxide to produce a clear aqueous solution, which was then lyophilized to give 980 mg of sodium ML-236A-carboxylate as a white powder.

Example 16 Sodium MB-530A-carboxylate 300 litres of a culture medium (pH 5.5 before sterilization) containing 5% w/v glucose, 0.5% w/v corn steep liquor, 2% w/v peptone ("Kyokuto" brand) and 0.5% w/v ammonium chloride were charged into a 600 litre fermenter and inoculated with the organism Monascus ruber SANK 18174. Cultivation was continued at 26"C for 11 6 hours under aerobic conditions at an aeration rate of 300 litres per minute and agitation at 1 90 rotations per minute. The culture broth (containing the organism) was adjusted to pH 3.4 with 6N hydrochloric acid and extracted with 800 litres of ethyl acetate. The extract was washed with 200 litres of a saturated aqueous solution of sodium chloride and dried over anhydrous sodium sulphate.It was then concentrated to 1 8 litres by evaporation under reduced pressure. The resulting solution was extracted with 60G litres of ethyl acetate. 50 g of trifluoroacetic acid were added to the extract and reacted at 800C for 30 minutes. The reaction mixture was washed successively with 10 litres of a 2% w/v aqueous solution of sodium bicarbonate and 10 litres of a 10% w/v aqueous solution of sodium chloride, and then was concentrated to give 105 g of an oily substance. This oil was dissolved in 400 ml of methanol. 20 ml of the methanolic solution (containing about 5.3 g of the oii) were subjected to preparative rapid liquid chromatography with a reverse phase column (the same as in Example 14). A 65:35 v/v mixture of methanol and water was used as the developer.The development was carried out at a flow rate of 200 ml/minute (developing time: about 1 5 minutes), watching a differential refractometer connected to the apparatus, the portion showing a main peak on the differential refractometer was separated. This operation was repeated and the resulting main peak fractions were collected and concentrated to give 7.9 g of an oily substance, which was dissolved in 30 ml of methanol. 60 ml of this methanolic solution (containing about 1.6 g of the oil) were subjected again to the same chromatography but developed with a 60:40 v/v mixture of methanol and water at a flow rate of 200 ml/minute. A portion showing a main peak was separated.

This operation was repeated and the main peak fractions were collected and concentrated. There were .obtained 679 mg of MB-530A as an oily substance. To this substance were added 21 ml of a 0.1 N aqueous solution of sodium hydroxide and the mixture was stirred at 50-600C for 3 hours. Insolubles were filtered off and the filtrate was lyophilized to give 720 mg of sodium MB-530A-carboxylate.

Elemental analysis: calculated for C9H290sNa: C, 63.33%; H, 8.06%.

Found: C, 63.51%: H, 7.97%.

Example 17 Sodium MB-530A-carboxylate 300 litres of a culture medium (pH 7.4 before sterilization) containing 1.5% w/v soluble starch, 1.5% w/v glycerine, 2% w/v fish meal and 0.2% w/v calcium carbonate were charged into a 600 litre fermenter and inoculated with the organism Monascus ruber SANK 17075 (CBS 832.70). Cultivation was continued at a temperature of 260C for 120 hours under aerobic conditions at an aeration rate of 300 litres per minute and agitation at 1 90 rotations per minute. The resulting culture broth was filtered using a filter press to give 35 kg of the wet organism, to which 1 00 litres of water were added.The mixture was adjusted to pH 1 2 with sodium hydroxide, with stirring, and left to stand at room temperature for 1 hour. 2 kg of Hyflo Super Cel filter aid were added to the mixture, which was filtered by a filter press. The filtrate was adjusted to pH 10 with hydrochloric acid and adsorbed on a column charged with 5 litres of HP-20 resin. It was then washed with 1 5 litres each of water and 10% v/v aqueous methanol, after which it was eluted with 60% v/v aqueous methanol. The eluate was concentrated to a volume of about 10 litres. The residue was adjusted to pH 2 with hydrochloric acid and extracted with ethyl acetate. The extract was washed with a saturated aqueous solution of sodium chloride, dried over anhydrous sodium sulphate and concentrated to dryness to give 20 g of an oily substance containing MB-530A carboxylic acid.To this oil was added methanol to a total volume of 100 ml. 20 ml of the solution were subjected to preparative rapid liquid chromatography equipped with a reversed phase column (the same as used in Example 14) and eluted with a 13% v/v aqueous methanol solution (containing 2% acetic acid) at a flow rate of 200 ml/minute. A portion showing a main peak on the differential refractometer was separated (7-10 minutes). The remaining 80 ml were treated in the same manner. The resulting main peak fractions were collected, concentrated and extracted with ethyl acetate. The extract was concentrated to dryness after adding heptane, to give 920 mg of an oily substance, which was dissolved in 20 ml of methanol and subjected to chromatography under the same conditions as above to give 120 mg of MB-530A carboxylic acid. To this product were added 2 ml of methanol and 100 ml of water and the resulting solution was adjusted to pH 8.0 with 1 N aqueous sodium hydroxide to produce a clear aqueous solution. This was lyophilized to give 110 mg of sodium MB-530A-carboxylate as a white powder.

Claims (54)

Claims

1. Compounds of formula (I):

wherein: R' represents a hydrogen atom or a methyl group; R2 represents a hydrogen atom, the alcoholic moietv of an ester of the cationic moietv of a salt: and R3, R4 and R5 are the same or different and each represents a hydrogen atom or an organic or inorganic acyl group, provided that, when R3 represents a 2-methylbutyryl group, R4 and R5 both represent acyl groups.

2. Compounds as claimed in Claim 1, in which: R1 represents a hydrogen atom or a methyl group; R2 represents a hydrogen atom, a metal atom, an ammonium group, an alkyl-substituted ammonium group, a group capable of forming a salt derived from a basic amino acid, an alkyl group, an aralkyl group optionally having a substituent on the aryl moiety or a phenacyl group optionally having a substituent on the phenyl moiety, said substituents being selected from C1-C4 alkyl groups, C1-C4 alkoxy groups, halogen atoms and trifluoromethyl groups; and R3, R4 and R5 are the same or different and each represents a hydrogen atom, an aliphatic acyl group, an aromatic acyl group or an araliphatic acyl group, provided that, when R3 represents a 2methylbutyryl group, R4 and R5 both represent acyl groups.

3. Compounds as claimed in Claim 1, in which: R1 represents a hydrogen atom or a methyl group; R2 represents a hydrogen atom, an alkali metal atom, an alkaline earth metal atom, aluminium, zinc, iron, germanium, an ammonium group, a group capable of for ming a salt derived from a basic amino acid, a C1-C6 alkyl group, an aralkyl group optionally having a substituent on the aryl moisty, or a phenacyl group optionally having a substituent on the phenyl moiety, said substituent being selected from C1-C4 alkyl groups, C1-C4 alkoxy groups, halogen atoms, and trifluoromethyl groups; and R , R4 and R5 are the same or different and each represents a hydrogen atom. a C2-C20 alkanoyl group, a C3-C20 alkenoyl group, a C3-C20 alkylnoyl group, a C7-C15 aromatic acyl group, a C8-C9 aralkanoyl group or a C9 aralkenoyl group, provided that, when R represents a 2-methylbutyryl group, R4 and R5 both represent acyl groups.

4. Compounds as claimed in Claim 1, in which: R1 represents a hydrogen atom or a methyl group; R2 represents a hydrogen atom, an alkali metal atom, an alkaline earth metal atom, an ammonium group, a group capable of forming a salt derived from a basic amino acid, a C1-C4 alkyl group, a benzyl group or a phenacyl group; and R3, R4 and R5 are the same or different and each represents a hydrogen atom, a C2-C6alkanoyl group, a C3-C6 alkenoyl group or a benzoyl group, provided that, when R3 represents a 2methylbutyryl group, R4 and R5 both represent acyl groups.

5. Compounds as claimed in Claim 1, in which: R1 represents a hydrogen atom or a methyl group; R represents a C1-C4 alkyl group, a benzyl group or a phenacyl group ; R represents a C2-C5 alkanoyl group or a C3-C6 alkenoyl group ; and R4 and R5 are the same or different and each represents a C2-C6 alkanoyl group or a benzoyl group.

6. Compounds as claimed in Claim 1, in which: R1 represents a hydrogen atom; R2 represents a C1-C4 alkyl group; R3 represents a C2-C6 alkanoyl group; and R4 and R5 are the same or different and each represents an acetyl group or a benzoyl group.

7. Compounds of formula (II) :

wherein: R1 represents a hydrogen atom or a methyl group; R28 represents a hydrogen atom, an alkali metal atom, an alkaline earth metal atom, an ammonium group, a group capable of forming a salt derived from a basic amino acid. a C1-C4 alkyl group. a benzyl group or a phenacyl group ; R3a represents a C2-C6 alkanoyl group other than a 2-methylbutyryl group or a C3-C0 alkenoyl group; and m represents the valence of R2a.

8. Compounds as claimed in Claim 7, in which R2a represents a C1-C4 alkyl group, a benzyl group or a phenacyl group.

9. Compounds as claimed in Claim 7. in which R28 represents an alkali metal atom.

10. Compounds of formula (II) :

wherein: R1 represents a hydrogen atom or a methyl group; and M represents a hydrogen atom or an alkali metal atom.

11. Compounds as claimed in Claim 10, in which M represents an alkali metal atom.

12. Compounds as claimed in Claim 10, in which M represents a sodium atom.

13. Sodium ML-236A-carboxylate.

14. Sodium MB-530A-carboxylate.

1 5. Sodium 8'-0-butyryl-ML-236A-carboxylate.

1 6. Sodium 8'-O-hexanoyl-ML-23 6A-carboxylate.

17. Ethyl 8'-O-butyryl-M L-23 6A-carboxylate.

1 8. Ethyl 8'-O-hexanoyl-ML-236A-carboxylate.

19. Butyl 8'-0-butyryl-ML-236A-carboxylate.

20. Butyl 8'-O-(4-peptenoyl)-ML-236A-carboxylate.

21. Butyl 8'-O-isovaleryl-M B-530A-carboxylate.

22. Benzyl 8'-O-butyryl-MB-236A-carboxylate.

23. Benzyl 8'-0-hexanoyl-ML-236A-carboxylate.

24. Phenacyl 8'-0-butyryl-MB-530A-carboxylate.

25. Butyl 3,5-di(O-acetyl)-ML-236B-carboxylate.

26. Butyl 3,5-di(O-benzoyl)-ML-236B-carboxylate.

27. A process for producing an ML-236A carboxylic acid salt in which an ML-236A-producing microorganism of the genus Penicillium is cultivated in a culture medium therefor and the ML-236A carboxylic acid salt is recovered from the resulting culture broth.

28. A process as claimed in Claim 27, in which said microorganism is a strain of Penicillium citrinum, Penicillium chrysogenun. or Penicillium notatum.

29. A process as claimed in Claim 28, in which said microorganism is Penicillium citrinum SANK 1 8767 (FERM 2609), Peniclllium citrinum SANK 24467, Peniclllium citrinum SANK 24567, Penicillium chrysogenum SANK 12768 (ATCC 100020) or Penicillium notatum SANK 24867.

30. A process as claimed in any one of Claims 27 to 29, in which said salt is an alkali metal salt.

31. A process for producing an MB-530A carboxylic acid salt in which an MB-530A- producing microorganism of the genus Monascus is cultivated in a culture medium therefor and the MB-530A carboxylic acid salt is recovered from the resulting culture broth.

32. A process as claimed in Claim 31, in which said microorganism is a strain of Monascus ruber Monascus anka, Monascus paxii Monascus purpureus or Monascus vitreus.

33. A process as claimed in Claim 32, in which said microorganism is Monascus ruber SANK 11272 (IFO 9203), Monascus ruber SANK 17075 (CBS 832.70), Monascus ruber SANK 17175 (CBS 503.70), Monascus ruber SANK 17275 (ATCC 181 99), Monascus ruber SANK 1 5177 (FERM 4956), Monascus ruber SANK 13778 (FERM 4959), Monascus ruber SANK 10671 (FERM 4958), Monascus ruber SANK 181 74 (FERM 4957), Monascus anka SANK 10171 (IFO 6540), Monascus paxii SANK 111 72 (IFO 8201). Monascus purpureus SANK 10271 (IFO 4513) or Monascus vitreus SANK 10960 (NIHS 609, e-609; FERM 4960).

34. A process as claimed in any one of Claims 31 to 33, in which said salt is an alkali metal salt.

35. A process for preparing a carboxylic acid salt of formula (IV):

wherein: R1 represents a hydrogen atom or a methyl group; R3 and R4 are the same or different and each represents a hydrogen atom or an acyl group, provided that, when R3 represents a 2-methylbutyryl group, R4 represents an acyl group; M' represents the cationic moiety of a salt; and m' is the valence of M1), in which a lactone compound of formula (V):

(wherein R', R3, R4, M1 and m' are as defined above) is hydrolyzed and, if desired, the carboxy group of the product thus obtained is converted to the corresponding salt.

36. A process as claimed in Claim 35, in which said hydrolysis is effected in an alkali solution and M' represents an alkali metal atom.

37. A process as claimed in Claim 35 or Claim 36, in which: R' represents a hydrogen atom or a methyl group: R3 represents a hydrogen atom, a C2-C6 alkanoyl group or a C3-C6 alkenoyl group; and R4 represents a hydrogen atom, a C2-C6 alkanoyl group or a benzoyl group.

38. A process for preparing a carboxylic acid derivative of formula (VI):

(wherein R' represents a hydrogen atom or a methyl group and M2 represents an alkali metal atom) in which a compound of formula (VII):

(wherein R1 is as defined above) is hydrolyzed in an alkali'solution.

39. A process as claimed in Claim 38, in which M2 represents a sodium atom and said alkali solution is an aqueous solution of sodium hydroxide.

40. A process for preparing a carboxylic acid ester of formula (Vlil):

(wherein: R1 represents a hydrogen atom or a methyl group; R3 and R4 are the same or different and each represents a hydrogen atom or an acyl group, provided that, when R3 represents a 2-methylbutyryl group, R4 represents an acyl group; and R2b represents an alkyl group or an aralkyl group), in which a lactone compound of formula (V):

(wherein Rt, R3 and R4 are as defined above) is reacted with an alcohol in the presence of an acid catalyst.

41. A process as claimed in Claim 40, in which said acid catalyst is an inorganic acid, a Lewis acid or a cation exchange resin.

GB 2 073 199 A

42. A process as claimed in Claim 41, in which said acid catalyst is a strongly acidic cation exchange resin.

43. A process as claimed in any one of Claims 40 to 42, in which: R' represents a hydrogen atom or a methyl group; R represents a hydrogen atom, a C2-C6 alkanoyl group or a C3-C6 alkenoyl group ; R4 represents a hydrogen atom, a C2-C6 alkanoyl group or a benzoyl group (provided that, when R3 represents a 2-methylbutyryl group, R4 represents an acyl group); and said alcohol is a C1-C4alkanol.

44. A process for preparing a carboxylic acid ester of formula (IX):

(wherein: R1, R3, H4 and R5 are as defined in Claim 1; R2C represents the alcoholic moiety of an ester; and p is the valence of R2c), in which a carboxylic acid salt of formula (X):

(wherein R1, R3, R4 and R5 are as defined above, M3.represents an alkali or alkaline earth metal atom, and q is the valence of M3) is reacted with an esterifying agent.

45. A process as claimed in Claim 44, in which said esterifying agent is an alkyl halide, an aralkyl halide or a phenacyl halide.

46. A process as claimed in Claim 44 or Claim 45, in which M3 represents an alkali metal atom.

47. A process as claimed in Claim 46, in which said salt (X) in which M3 represents an alkali metal atom is formed in situ prior to said reaction with an esterifying agent.

48. A process as claimed in any one of Claims 44 to 47, in which: R' represents a hydrogen atom or a methyl group; R3 represents a hydrogen atom, a C2-C6 alkanoyl group or a C3-C6 alkenoyl group; R4 and R5 are the same or different and each represents a hydrogen atom, a C2-C6 alkanoyl group or a benzoyl group, provided that, when R3 represents a 2-methylbutyryl group, R4 and R5 both represent acyl groups; M3 represents an alkali metal atom; and said esterifying agent is a benzyl halide or a phenacyl halide.

49. A process for preparing a triacyl compound of formula (Xl):

(wherein: R', R2 and n are as defined in Claim 1; and R3b, R4b and R5b are the same or different and each represents an acyi group), in which a compound of formula (XII):

(wherein R1, R2 and n are as defined above and R3C, R4C and R5C are the same or different and each represents a hydrogen atom or an acyl group provided that at least one of R3C, R4C and R5C represents a hydrogen atom) is acylated.

50. A process for preparing one or more compounds of formulae (XIII), (XIV) and (XV):

(wherein: R1, R2, H4 and n are as defined in Claim 1; R3b represents an acyl group; and R4b represents an acyl group), in which a compound of formula (XVI):

(wherein R1, R2, R3b and n are as defined above) is acylated to produce compounds of said formulae (XIII), (XIV) or (XV) and they are isolated from the reaction mixture.

51. A process as claimed in Claim 50, in which, in said compound (XVI): R' represents a hydrogen atom or a methyl group; R2 represents an alkali metal atom, a C1-C4 alkyl group, a benzyl group or a phenacyl group; and R3b represents a C2-C6 alkanoyl group or a C3-C6 alkenoyl group, said acylation is effected with a C2-C8 alkanoic acid, benzoic acid or a reactive derivative thereof, and there is produced said compounds (Xlil) and/or (XIV) in which R4b represents an alkanoyl group or a benzoyl group.

52. A process as claimed in Claim 50, in which said acylation is effected with a reactive derivative of a carboxylic or sulphuric acid.

53. A process as claimed in Claim 52, in which said acylation is effected with an acid halide, an acid anhydride or a mixed anhydride of a carboxylic or sulphonic acid.

54. A process for preparing compounds of formula (I), as defined in Claim 1, substantially as hereinbefore described with reference to any one of the foregoing Examples.