IE904375A1 - Substituted 10-membered ring lactones, processes for their¹preparation use thereof - Google Patents

Abstract

Novel 10-membered ring lactone derivatives of the formulae I and II a process for the preparation of these compounds, pharmaceutical compositions which contain the active compounds according to the invention and their use as medicaments having, in particular, hypolipidaemic action are described.

Description

Description Substituted 10-membered ring lactones, processes for their preparation and use thereof The invention relates to substituted 10-membered ring lactones, processes for their preparation and their use, in particular as pharmaceuticals which inhibit cholesterol biosynthesis. The compounds according to the invention may accordingly be employed as hypolipidemics. -membered ring lactones of the formulae have already been proposed in European Patent Application 89104160.0. They exhibit antibacterial action. These compounds are obtained by culturing microorganisms of the species Penicillium, in particular Penicillium Species DSM 4209 and DSM 4210, in a nutrient medium.

It has now been found that certain derivatives of these 10-ring lactones effectively inhibit cholesterol biosynthesis. The invention thus relates to: -ring lactones of the formulae I and II R1 (II) in which A1 and A2 are identical or different and are H or Ci-C*alkyl, the Ci-C^-alkyl radical being unsubstituted, or monosubstituted or polysubstituted by OH or phenyl, the phenyl radical in turn being unsubstituted, or monosubstituted or polysubstituted by halogen, CF3 or C1-C3-alkyl or in which A1 and A2 together form a radical of the formula or A1 or in which A3 and A3’ are identical or different and are C1-C2-alkoxycarbonyl, phenyl or Cj-C^-alkyl, in which and A2 are together a radical of the formula A in which A* is phenyl or Cx-Cg-alkyl, A* R1 is -H, =0 or -OR3 and R2 is =0, -OR4, =N-R5 or , where in the case of the formula II for R2 = eN-R5 and Α1 β H, this can also be present as the compound of the formula II' (II') in which R3 is H, Cv-Ce-alkyl, benzyl, allyl, MEM, MOM, SEM, THP, 4'-methoxytetrahydropyran-4'-yl, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, dimethylcyclohexylsilyl, dimethyl tertiary-butylsilyl or a sulfonic acid ester of the formula -SO2R7 in which R7 is Ci-Cuj-alkyl, phenyl or p-methylphenyl or R3 is a mono- or di saccharide whose OH groups are unprotected or are protected by protecting groups or X X R3 is a radical of the formula -C-Z-R8 or -£-Z-(CH2)n-R8 in which X is O or S Z is 0, -N-R6, -N-(CH2)n-R8 or -NH n is 1 to 3 and R8 is Cj-Cg-alkyl, C3-Ce-cycloalkyl, aryl, pyridyl, pyrimidyl or pyrazinyl or ί .

R is a radical of the formula -C-R in which R9 is C1-C20-alkyl, C2-C16-alkenyl having 1-3 double bonds, C2-C16-alkynyl having 1-3 triple bonds, C3-C9-cycloalkyl, aryl or heteroaryl, the alkyl, alkenyl, alkynyl and cycloalkyl radicals being unsubstituted, or monosubstituted or polysubstituted by COOA5, halogen, aryl, aryloxy, heteroaryl, heteroaryloxy, NHCOCH3, COA5 or SO3A5 in which A5 is C1-C4-alkyl or H and in which all said aryl, heteroaryl, pyridyl, pyrimidyl and pyrazinyl radicals are unsubstituted, or monosubstituted or polysubstituted by halogen, nitro, cyano, hydroxyl, trifluoromethyl, amino, carboxyl, Cx-C4-alkylcarbonyl, Cj-C^-alkoxy30 carbonyl, Cx-Cx0-alkyl, Cx-C4-alkoxy or NHCOCHa and in which R*, independently of R3, has the meanings indicated above for R3 and R5 and R5* are identical or different and are hydrogen, Cx-Cxo-alkyl, C2-C10 -alkenyl, benzyl, phenyl or phenylethyl, said radicals being unsubstituted or substituted by hydroxyl, halogen, NH2 or OCH3, and the physiologically tolerable salts, excluding the compound of the formula I in which R2 is - 4 -OR* where R* = H and also excluding the compound of the formula II in which R1 is -OR3 and R2 is =0 where R3 = H.

Preferred 10-membered ring lactones of the formulae I and II as indicated above are those in which A1 and A2 are identical or different and are H or Cx-C2alkyl, the Cx-C2-alkyl radical being unsubstituted or monosubstituted by phenyl, the phenyl radical in turn being unsubstituted or monosubstituted by halogen, CF3 or Cx-C3-alkyl or in which A3 A1 and A2 together are a radical of the formula zztz A3 in which A3 and A3* are identical or different and are Cx-C2-alkoxycarbonyl or phenyl or in which A1 and A2 together are a radical of the formula in which A* is phenyl, R1 is -H, =0 or -OR3 and ^R5 R2 is =0, -OR4, =N-R5 or ^R5' · where in the case of the formula II for R2 = =N-R5 and A1 = H, this can also be present as the compound of the formula II’, as indicated above, in which R3 is H, Cx-C8-alkyl, benzyl, allyl, MEM, MOM, SEM, THP, 4'-methoxytetrahydropyran-4·-yl, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, dimethylcyclohexylsilyl, dimethyl tertiary-butylsilyl or a sulfonic acid ester of the formula -SO2R7 in which R7 is methyl, phenyl or p-methylphenyl or R3 is a pyranosyl radical whose OH groups are unprotected or protected by protecting groups or X X R3 is a radical of the formula -(5-Z-R8 or -C-Z-(CH2)n-R8 in which X is 0, I - 1 β 1 Z is O, -N-R8, -N-(CH2)n-R® or -NH n is 1 to 3 and R® is Cj-Ca-alkyl, C3-C6-cycloalkyl or aryl or O Η β R is a radical of the formula -C-R in which Re is Cx-Cu-alkyl, C2-C16-alkenyl, C3-C6-alkynyl, C3-C6-cycloalkyl, aryl or heteroaryl, the alkyl, alkenyl, alkynyl and cycloalkyl radicals being unsubstituted, or monosubstituted or polysubstituted by COOA5, halogen, phenyl, phenyloxy, NHCOCHj, COA5 or SO3A5 A5 being Cx-C^-alkyl or H and in which all said aryl and heteroaryl radicals are unsubstituted, or monosubstituted or polysubstituted by halogen, cyano, trifluoromethyl, carboxyl, Cx-Ckalkylcarbonyl, Ci-C^-alkoxycarbonyl, Ci-Cj-alkyl, Ci-C4-alkoxy or NHCOCH3 and in which R*, independently of R3, has the meanings indicated above for R3 and R5 and R5' are identical or different and are hydrogen, Cx-Cnj-alkyl, C2-C10-alkenyl, benzyl, phenyl or phenylethyl, said radicals being unsubstituted or substituted by hydroxyl, halogen, NH2 or OCH3, and the physiologically tolerable salts, excluding the compound of the formula I in which R2 is -OR4 where R4 = H and also excluding the compound of the formula II in which R1 is -OR3 and R2 is =0 where R3 = H.

Particularly preferred 10-membered ring lactones of the formulae I and II as indicated above are those in which A1 and A2 are identical or different and are H or methyl, the methyl radical being unsubstituted or monosubstituted by phenyl, or in which A1 radical of the formula and A2 together are a in which A3 and A3’ are identical and are ethoxycarbonyl R1 is -H, =0 or -OR3 and -Ν' R5 is =0, -OR4, “N-R5 or R5 , where in the case of the formula II for R2 - “N-R5 and A1 H, this can also be present as the compound of the formula II', as indicated above, in which R3 is H, methyl, benzyl, THP, trimethylsilyl, dimethyl tertiary-butylsilyl or a sulfonic acid ester of the formula -SO2R7 in which R7 is p-methylphenyl or R3 is a glucosyl, galactosyl or lactosyl radical whose OH groups are unprotected or are protected by acetyl groups or benzyl groups, these said sugar radicals either being glycosidically bonded or linked via an orthoester or R3 X X is a radical of the formula -C-Z-R8 or -2)n-R8 in which X is 0, Z is -NH n is 1 and R8 is C4-C8-alkyl, or aryl or O R3 is a radical of the formula -<^-R9 in which R9 is Cj-Cjs-alkyl, C2-C8-alkenyl, C3-C6-cycloalkyl, phenyl, thienyl or furyl, the alkyl, alkenyl, and cycloalkyl radicals being unsubstituted, or monosubetituted or polysubstituted by COOA5, F, Cl, Br, phenyl, phenyloxy, NHCOCH3, or COA3, A5 being methyl or H and in which all said aryl or phenyl radicals are unsubstituted, or monosubstituted or polysubstituted by F, Cl, Br, trifluoromethyl, carboxyl, Ci-C^-alkylcarbonyl, Cx-C^-alkoxycarbonyl, Cx-Cj-alkyl, methoxy or NHCOCH3 and in which R4, independently of R3, has the meanings indicated above for R3 and R5 and R3’ are identical or different and are hydrogen, Cx-Cg-alkyl, benzyl, phenyl or phenylethyl, said radicals being unsubstituted or substituted by F, Cl, Br or OCH3, and the physiologically tolerable salts, excluding the compound of the formula I in which R2 is -OR4 where R4 = H and also excluding the compound of the formula II in which R1 is -OR3 and R2 is =0 where R3 = H.

Aryl is understood as meaning aromatic hydrocarbons, in particular phenyl and naphthyl, and heteroaryl as meaning heteroaromatic hydrocarbons, in particular thiophene and furan, but also pyridine, pyrimidine and pyrazine.

Halogens are understood as meaning fluorine, chlorine, bromine and iodine, THP as meaning tetrahydropyranyl, MEM as meaning methoxyethoxymethyl, MOM as meaning methoxymethyl and SEM as meaning jS-trimethylsilylethoxymethyl.

All said alkyl and alkenyl radicals having more than 2 carbon atoms and all alkynyl radicals having more than 3 carbon atoms can be both straight-chain and branched. Said alkenyl and alkynyl radicals are both monounsaturated and polyunsaturated compounds, preferably monouns aturated. - 8 Said polysubstituted aryl, heteroaryl, pyridyl, pyrimidyl and pyrazinyl radicals are 2-, 3- or, if possible, tetra- or pentasubstituted radicals, it being possible for 2 or more substituents to be identical or different.

In said polysubstituted alkyl, alkenyl, alkynyl and cycloalkyl radicals, two to all of the replaceable hydrogen atoms can be replaced by said substituents. Preferably, the abovementioned polysubstituted radicals are di- or trisubstituted radicals. The substituents can be identical or different.

Said sugar radicals are mono- and disaccharides such as are indicated, for example, in the Biochemischen Taschenbuch (Biochemical Pocket Book), 2nd edition, 1st part, pages 107-177, Springer-Verlag, Berlin, Gdttingen, Heidelberg, 1964. In particular, they are hexopyranosides and hexofuranosides; glucose, fructose, allose, altrose, mannose, lactose and galactose are very particularly preferred. The OH groups present in said sugar radicals may optionally be protected by the protecting groups customary in sugar chemistry such as acetyl, benzoyl, benzyl or acetonyl. The sugar radical can be bonded glycosidically or via an orthoester.

The invention furthermore relates to a process for the preparation of the compounds of the formulae I and II in which a) the compound of the formula III is reacted with a compound of the formula IV R9 (IV) Ο II c in which Q is OH, chloride, bromide, imidazolide or an acid anhydride and R9 has the meanings mentioned above for formula I and II, or in which b) the compound of the formula III is reacted with a compound selected from amongst: dihydropyran, 4-methoxy-5,6-dihydropyran, C]-Cealkyl, benzyl or allyl chloride, bromide or iodide or MEM, MOM or SEM chloride, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, dimethylcyclohexyl si lyl or dimethyl tertiary-butylsilyl chloride, sulfonic acid halides of the formula Hal-SO2R7 in which Hal is chlorine, bromine or iodine and R7 has the abovementioned meanings, isocyanates of the formula = C = N-R8 or 0 = C = N-(CH2)n-R8, isothiocyanates of the formula S = C = N-R® or S = C = N-(CH2)n-R8, carbamoyl halides of the formula Hal' -CO-N- (R8) 2 or Hal'-CO-N-[ (CH2)n-R®]2 thiocarbamoyl halides of the fornula Hal'-CS-N-(R8)2 or Hal'-CS-N-[ (CH2)n-R®)2 or compounds of the formulae Hal'-C(O)-O-R®, Hal ’-CO-(CH2) n-R®, Hal'-CS-O-R® or Hal'-CS-O-(CH2)n-R® in which n and R® have the meanings mentioned above for formula I and II and Hal' is chlorine or bromine, or in which c) the compound of the formula III is reacted with a compound of the formula V Η - (V) in which R5 and Rs* are defined as indicated above for formula I and II, and the resulting C-C double bond in the compound of the formula II' is option5 ally hydrogenated, or in which d) the compound of the formula III is reacted with a compound of the formula VI R3 - Hal (VI) in which R3 is a mono- or disaccharide and Hal is as defined above, or in which e) the compound of the formula III is oxidized to a compound of the formula VII CH, o a 0“^ 0 (VII) or in which f) the compound of the formula III is reduced to a compound of the formula or in which g) a compound of the formula III is subjected to elimination to give the enone and the C-C double bond is a compound of the formula optionally hydrogenated, being formed, or in which h) the compound of the formula III is reacted with a compound of the formula VIII and/or VIII' A1 - Abg (VIII) A2 - Abg (VIII') in which A1 and A2 are as defined in formula I and II f ^A3 except for the meaning =£?? and Abg is ^A3 chlorine, bromine, iodine, sulfate, mesylate or tosylate or in which i) the compound of the formula III is reacted with a compound of the formula IX (IX) in which A3 and A3' are as defined above for formula I and II, or in which j) the compound of the formula III is reacted with a compound of the formula X H-^A4 (X) in which A4 is as defined above for formula I and II or in which k) the compound of the formula III is reacted with diketene a compound of the formula II being formed where r1 = 0-C(O)-CH2-C(O)-CH3 or in which 1) a compound of the formula III is reacted with a mesoxalic acid diester, a compound of the formula II A3 being formed where A1 and A2 = =? in which A3 A3 and A3' are as defined in formula I and II, or in which two or more of the abovementioned process variants a-1 are carried out successively and in which the compounds I or II are then optionally converted into their physiologically tolerable salts.

The compound of the formula III necessary for the pre25 paration of compounds of the formula I can be prepared, for example, by the procedure proposed in German Patent Application P 3808492.9 - to which reference is expressly made here. In this process, the compound of the formula III is produced from Penicillium Species, preferably DSM 4209 or DSM 4210, in a nutrient solution which contains a source of carbon and a source of nitrogen and also customary inorganic salts. Instead of the strain DSM 4209 or DSM 4210, of course, their mutants and variants can also be employed if they synthesize this compound.

The formation of the compound of the formula III proceeds particularly well in a nutrient solution which contains about 0.2 to 5%, preferably 1 to 4%, of malt extract, 0.02 to 0.5%, preferably 0.1 to 0.4% yeast extract, 0.1 to 5%, preferably 0.5 to 2% glucose and 0.005 to 0.2%, preferably 0.01 to 0.1% ammonium salt, in each case relative to the weight of the total nutrient solution.

Culturing is carried out aerobically, ie. for example in submerse fashion with shaking or stirring in shaker bottles or fermenters, if desired with introduction of air or oxygen. It can be carried out in a temperature range from about 18 to 35eC, preferably at about 25 to °C, in particular at 27 to 28’C. The pH range should be between 6 and 8, preferably between 6.5 and 7.5. The microorganism is in general cultured under these conditions over a period of 60 to 170 hours, preferably 100 to 150 hours.

To isolate the compound, culture broth, and mycelium are first extracted with organic solvents, such as, for example, chloroform, ethyl acetate, etc., in order to remove the non-polar impurities. The mixture is then extracted with a polar solvent, for example lower alkanols or mixtures of chloroform and/or ethyl acetate with a lower alkanol.

Isolation in pure form is preferably carried out on suitable media, such as, for example, silica gel, alumina or ion exchangers, by means of subsequent elution with organic polar solvents or solvent mixtures.

In the following, processes a) to 1), which enable the preparation of the differently substituted 10-ring lactones, are described in more detail.

In process variant a), a procedure is best used in which 5 the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert aprotic solvent such as chloroform, methylene chloride, tetrahydrofuran (THF), ethyl acetate or dioxane, with a compound of the formula IV until the reaction is complete, if appropriate in the presence of a base, preferably pyridine or triethylamine. If a compound of the formula IV where Q=OH is employed, the addition of dicyclohexylcarbodiimide, if appropriate in the presence of a catalyst such as DMAP, is recommended.

The reaction temperatures here are between -70 “C and +100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70 eC and +40*C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. The completion of the reaction can be determined, for example, by means of thin layer chromatography (TLC checking).

If not commercially available, the starting compounds for process variant a), which are compounds of the formula IV, can be prepared in a simple manner by methods known from the literature. For example, the acid chlorides are obtained by reaction of the corresponding carboxylic acid with thionyl chloride, PC13 or PC15.

In process variant b), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, with a compound which is selected from amongst: dihydropyran, 4-methoxy-5,6-dihydropyran, Cx-Ce-alkyl, benzyl or allyl chloride, bromide or iodide or MEM, MOM, or SEM chloride, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, dimethylcyclohexylsilyl or dimethyl tert.fc904375 butylsilyl chloride, sulfonyl halides of the formula Hal-SO2R7, isocyanates of the formula O=C=N-R8 or O=C=N-(CH2)n-R8, isothiocyantes of the formula S=C=N-R8 or S=C=N- (CH2)n-R8, carbamoyl halides of the formula Hal'-CO-N-(R8) 2 or Hal '-CO-N-[ (CH2)n-R8]2, thiocarbamoyl halides of the formula Hal'-CS-N-(R8)2 or Hal'-CS-N-[ (CH2)n-R8]2, or compounds of the formulae Hal'-C(O)-O-R8, Hal'-CO-(CH2)n-R8, Hal'-CS-O-R8 or Hal'-CS-O-(CH2)n-R8.

If desired, this reaction can also be carried out with the addition of base. Examples of suitable bases are triethylamine, pyridine or lutidine. In the case of the reaction with dihydropyran and 4-methoxy-5,6-dihydropyran, acid catalyst such as H2SO4, HCl, p-toluenesulfonic acid or pyridinium p-toluenesulfonate can be employed. A variant of the process b) comprises working in a suitable, preferably inert, solvent such as chloroform, methylene chloride, THF, ethyl acetate or dioxane. Here too, the excess of the abovementioned compounds can be up to a 50-fold amount.

The reaction temperatures here are between -70“C and +100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70*C and -40°C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of TLC checking.

If not commercially available, the starting compounds for process variant b), can be prepared in a simple manner by methods known from the literature. For example, the sulfonyl halides of the formula Hal-S02R7 are obtained by free radical reaction of alkanes with chlorine and S02 or by halogenation of aromatics with halosulfonic acid Hal-SO3H. The isocyanates, isothiocyanates, carbamoyl halides and thiocarbamoyl halides are obtained by methods - 16 known from the literature.

In process variant c), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert solvent such as chloroform, methylene chloride, tetrahydrofuran (THF), toluene or hexane, with a compound of the formula V until the reaction is complete, if appropriate in the presence of a catalyst, preferably pyridinium p-toluenesulfonate or p-toluene10 sulfonic acid, if appropriate in the presence of a dehydrating agent such as molecular sieve or by means of azeotropic distillation.

The reaction temperatures here are between -70‘C and +100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70°C and +40C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 12 hours.

Completion of the reaction can be determined, for example, by means of thin-layer chromatography.

If not commercially available, the starting compounds for process variant c), which are compounds of the formula V, can be prepared in a simple manner by methods known from the literature.

In process variant d), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert solvent such as chloroform, methylene chloride, nitromethane, acetonitrile or toluene, with a compound of the formula VI until the reaction is complete, in the presence of a catalyst such as tetraethylammonium bromide/molecular sieve 4A, Hg(CN)2, Ag2CO3, AgC104 or silver trifluoromethanesulfonate, if appropriate in the presence of a base, such as collidine, tetramethylurea.

The reaction temperatures here are between -7O’C and +100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70 *C and + 40 ®C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

If not commercially available, the starting compounds for process variant d), which are compounds of the formula VI, can be prepared in a simple manner by methods known from the literature. Mono- or disaccharides substituted by halogen in the 2-position are preferred.

In process variant e), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert solvent such as chloroform, carbon tetrachloride, methylene chloride or hexane, with an oxidant such as pyridinium chlorochromate or pyridinium dichromate until the reaction is complete.

The reaction temperatures here are between -70’C and +100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70’C and +40*C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

Oxidants for process variant e) are commercially available.

In process variant f), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert solvent such as diethyl ether, THF or lower alcohols, with a reductant such as sodium borohydride, sodium cyanoborohydride or lithium aluminum hydride until the reaction is complete.

The reaction temperatures here are between -70 °C and +100eC, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70’C and +40’C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

The reductants for process variant f) are commercially available.

In process variant g), the compound of the formula III is first converted into a corresponding O-mesylate or O-tosylate according to process b) and this is then subjected to elimination by means of base by methods known from the literature. The elimination product can then be hydrogenated by methods known from the literature. In this case, the common hydrogenation catalysts such as palladium or platinum catalysts can be used.

The reaction temperatures here are between -70*C and +100 °C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70*C and +40*C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

In process variant h), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate ,E 904375 in an inert solvent such as chloroform, methylene chloride, tetrahydrofuran (THF), ethyl acetate or dioxane, with a compound of the formula VIII and/or VIII' until the reaction is complete.

The reaction temperatures here are between -70°C and +100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70 *C and +40’C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

If not commercially available, the starting compounds for process variant h), which are compounds of the formula VIII or VIII', can be prepared in a simple manner by methods known from the literature.

In process variant i), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert aprotic solvent such as chloroform, methylene chloride, tetrahydrofuran (THF), ethyl acetate or dioxane, with a compound of the formula IX until the reaction is complete, if appropriate in the presence of a base such as pyridine or piperidine.

The reaction temperatures here are between -70 *C and +100’C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70*C and +4O’C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

If not commercially available, the starting compounds for process variant i), which are compounds of the formula IX, can be prepared in a simple manner by methods known from the literature.

In process variant j), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert solvent such as tetrahydrofuran (THF) or diethyl ether, with a compound of the formula X until the reaction is complete, if appropriate in the presence of a base, preferably pyridine or piperidine.

The reaction temperatures here are between -7O’C and + 100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70’C and +40‘C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

If not commercially available, the starting compounds for process variant j), which are compounds of the formula X, can be prepared in a simple manner by methods known from the literature.

In process variant k), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert solvent such as chloroform, methylene chloride, tetrahydrofuran (THF), ethyl acetate or dioxane, with diketene until the reaction is complete, if appropriate in the presence of a base such as pyridine or triethylamine.

The reaction temperatures here are between -70eC and +100’C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70’C and +40’C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

The starting compound diketene for process variant k) is 5 commercially available.

In process variant 1), a procedure is best used in which the compound of the formula III is reacted in equimolar amounts or in an excess of up to 50-fold, if appropriate in an inert solvent euch as chloroform, methylene chloride, tetrahydrofuran (THF), ethyl acetate or dioxane and if appropriate with the addition of a base such as pyridine or piperidine, with a mesoxalic acid diester until the reaction is complete.

The reaction temperatures here are between -70“C and +100°C, when using a solvent preferably between the solidification point and the boiling point of the solvent, in particular between -70eC and +40*C. The reaction times are 1 to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8 hours. Completion of the reaction can be determined, for example, by means of thin layer chromatography.

If not commercially available, the starting compounds for process variant 1), which are mesoxalic acid diesters, can be prepared in a simple manner by methods known from the literature.

Not only the compound of the formula III, but also a compound which has already been derivatized by one or more of processes a )-1), can be used as the starting substance for the reactions according to process variants a)-l). In this manner, all types of substituents A1, A2, R1 and/or R2 can be successively introduced into the basic structure of the formula I or II.

The substances are purified, isolated and worked up by the customary methods; for example, the reaction products can be purified by chromatography on polar support materials such as silica gel or ’Sephadex LH 20 using solvents such as lower alkanols, such as methanol, or chloroform, dichloromethane or ethyl acetate or methanol/chloroform mixtures or chloroform/hexane/methanol mixtures but also by extractive methods such as liguid/liguid extraction or solid/liquid extraction or by crystallization.

The inhibition of cholesterol biosynthesis by the compounds according to the invention and their physiologically tolerable salts including the compound of the formula I in which R2 is -OR* where R* = H, and including the compound of the formula II in which R1 is -OR3 and R2 is =0 where R3 = H was checked in in vitro and in vivo tests. In these tests, it was shown that the derivatives according to the invention exhibited an excellent action as inhibitors of cholesterol biosynthesis. They can accordingly be employed as hypolipidemics.

Owing to their pharmacological properties, the compounds according to the invention are suitable for the treatment of disturbances of the metabolism of cholesterol and cholesterol-like substances.

The invention therefore further relates to the use of the compounds of the formulae I and II according to the invention and their physiologically tolerable salts for the treatment and prophylaxis of disorders of the metabolism of cholesterol and cholesterol-like substances.

The compounds can be used as pharmaceuticals either alone or mixed with physiologically tolerable auxiliaries or excipients. For this purpose, they can be administered orally in doses of 0.01-5.0 mg/kg/day, preferably 0.01 - 1.0 mg/kg/day or parenterally (subcutaneously) in doses ' of 0.001 - 2.5 mg/kg/day, preferably 0.001 - 1.0 mg/kg/day, in particular 0.005-0.2 mg/kg/day.

In severe cases, the dosage can also be increased. However, in many cases lower doses are also sufficient. These data relate to an adult of about 75 kg weight.

The invention furthermore includes the use of the com5 pounds according to the invention for the preparation of pharmaceuticals which are employed for the treatment and prophylaxis of the abovementioned diseases.

The invention further relates to pharmaceuticals which contain one or more compounds of the formula I and/or II according to the invention and/or their physiologically tolerable salts.

The pharmaceuticals are prepared by processes which are know per se and familiar to the person skilled in the art. The pharmacologically active compounds according to the invention (= active compound) are either used as pharmaceuticals as such or preferably in combination with suitable pharmaceutical auxiliaries or excipients in the form of tablets, coated tablets, capsules, suppositories, emulsions, suspensions or solutions, the active compound content being up to about 95%, advantageously between 10 and 75%.

In addition to solvents, gel-forming agents, suppository bases, tablet auxiliaries and other active compound carriers, examples of suitable auxiliaries or excipients for the desired pharmaceutical formulation are also antioxidants, dispersants, emulsifiers, defoamers, flavor correctants, preservatives, solubilizers or colorants.

The active compounds can be administered orally, parenterally (subcutaneously) or rectally.

The active compounds are mixed with the additives suitable for this purpose such as excipients, stabilizers or inert diluents and brought into suitable forms for administration, such as tablets, coated tablets, hard - 24 gelatine capsules, agueous alcoholic or oily suspensions or aqueous or oily solutions, by the customary methods. Examples of inert excipients which can be used are gum arabic, magnesia, magnesium carbonate, potassium phos5 phate, lactose, glucose or starch, in particular cornstarch. Preparation can be carried out here both as dry and as moist granules. Examples of possible oily excipients or solvents are vegetable or animal oils, such as sunflower oil or cod liver oil.

For subcutaneous or intravenous administration, the active compounds, if desired with the substances suitable for this purpose such as solubilizers, emulsifiers or other auxiliaries, are brought into solution, suspension or emulsion. Examples of suitable solvents are physio15 logical saline solution or alcohols, for example ethanol, propanol and glycerol, in addition also sugar solutions such as glucose or mannitol solutions, or alternatively a mixture of all types of the solvents mentioned.

The invention is illustrated in more detail below by means of examples.

All compounds obtained according to the following examples were characterized by means of 1H-NMR and/or 13C-NMR and/or IR and/or C,H analysis and/or mass spectrum. The 10-ring lactone of the formula III was prepared according to German Patent Application P 3808492.9.

Preparation of the compound of the formula III a) Preparation of epore suspension of the producer strain: 100 ml of nutrient solution (2 g of yeast extract, 20 g of malt extract, 10 g of glucose, 0.5 g of (NH4)2PO4, 1 1 of tap water, pH before sterilization 7.3) in a 500 ml Erlenmeyer flask are inoculated - 25 with the strain DSM 4209 or DSM 4210 and incubated at 25 °C and 120 rpm on a rotating shaker for 72 hours. 20 ml of culture fluid were then uniformly distributed in a 500 ml Erlenmeyer flask containing the nutrient base of the abovementioned composition, to which 20 g of agar/1 had been added for solidification, and decanted. The cultures are incubated at 25°C for 10 to 14 days. The spores from one flask formed after this time are washed using 500 ml of deionized water which contains one drop of a commercial nonionic surfactant (Triton X 100, Serve), and immediately reused or stored at -22’C. b) Production of a culture or pre-culture of the producer strain in an Erlenmeyer flask.

A 500 ml Erlenmeyer flask containing 100 ml of the nutrient solution described under a) is inoculated with a culture grown in a slant tube or with 0.2 ml of spore suspension and incubated at 120 rpm and 25°C on a shaker. The maximum production is achieved after about 120 hours. A submerse culture (5%) 48 hours old from the same nutrient solution is sufficient for inoculating 10 and 100 1 fermenters. c) Preparation of the compound of the formula III A 10 1 fermenter is operated under the following conditions: Nutrient medium 20 g/1 of malt extract 2 g/i of yeast extract 10 g/i of glucose 0.5 g/i of (NH4)2PO4 pH 7 .2 Incubation time: 150 hours Incubation temperature: 25’C Stirrer velocity: 250 rpm Aeration: 4 1 of air/min.

Development of froth can be suppressed by repeated addition of a few drops of ethanolic polyol solution. The production maximum is achieved after about 150 hours. The yields are about 100 mg/1. d) Isolation of the compound of the formula III After the fermentation of DSM 4209 or DSM 4210, the culture broth is filtered with the addition of 2% Celite as a filter aid. The compound of the formula III is isolated from the culture supernatant. It is processed according to the following scheme: Working-up/isolation Abbreviations: MPLC: Medium NP: Normal pressure liquid chromatography pressure column chromatography Example 1: Preparation of 6-butylcarbonyloxy-2,4-dioxo-10-methyl-loxa-7-cyclodecene oxide (II.11) 100 mg of the ten-ring lactone of the formula III 5 (0.47 mmol) are dissolved in 3 ml of dichloromethane (absolute) and stirred with 0.4 ml of valeric anhydride 42.5 mg (0.54 mmol) of abs. pyridine (15% excess) and a spatula tip full of N,N-dimethylaminopyridine at room temperature for 18 h. The solvent is then removed in vacuo and the oil remaining is purified by flash chromatography on silica gel (mobile phase chloroform).

Yield: 118.5 mg (0.40 mmol; 85% of theory), colorless oil C15H22O6: calc.: C 60.3 H 7.3 found: C 60.4 H 7.4 Example 2: Preparation of 2,4-dioxo-10-methyl-l-oxa-6-(phenylaminocarbonyloxy)-7-cyclodecene oxide (II.4) 214 mg of the ten-ring lactone of the formula III (1 mmol) are dissolved in 2.5 ml of absolute pyridine and stirred with 125 mg (1.05 mmol) of phenyl isocyanate at room temperature for 12 h. For working-up, the reaction mixture is concentrated in vacuo, the residue is taken up using dichloromethane, and the organic phase is washed with water, dried over Na2SO4 and concentrated in vacuo.

The oil remaining is purified by means of flash chromatography on silica gel (mobile phase: CHCl3/MeOH = 40:1).

Yield: 166 mg (0.50 mmol; 50% of theory); colorless crystals of melting point 149’C C17H19NO6: calc.: C 61.1 H 5.6 found: C 61.3 H 5.8 - 29 Example 3 Preparation of 10-methyl-l-oxa-2,4,6-trioxo-7-cyclodecene oxide (11.19) mg of the ten-ring lactone of the formula III 5 (0.23 mmol) are dissolved in 2.5 ml of dichloromethane and 75.5 mg (1.5-fold excess) of pyridinium chlorochromate (PCC) are added at a temperature of O’C. The reaction mixture is stirred at this temperature for 3 h.

The mixture is then filtered through filter aid and the organic phase is washed twice with water. After drying over Na2SO4 and removing the solvent in vacuo, a yellow oil remains which is purified by means of flash chromatography on silica gel (mobile phase: CHCl3/hexane/methanol = 8:8:1).

Yield: 26.5 mg (0.13 mmol; 54% of theory); colorless oil C10H12O5: calc.: C 56.8 H 5.6 found: C 56.6 H 5.7 nC-NMR spectrum (90.5 MHz; CDC13/TMS): δ = 192.53 (C-4); 192.19 (C-6); 164.69 (C-2); 69.10 (C-10); 56.74 (C-7); 56.07 (C-8); 55.98 (C-3); 52.05 (C-5); 33.60 (C-9); 20.57 (C-ll).

Example 4: Preparation of 4-hexylamino-6-hydroxy-10-methyl-l-oxa-3cyclodec-7-ene oxide (II'.1) 50 mg of the ten-ring lactone of the formula III are dissolved in 2 ml of absolute tetrahydrofuran and 2.0 equivalents of n-hexylamine (48 mg) are added. The reaction mixture is stirred at room temperature for 3 days.

For working-up, the mixture is freed from solvent in vacuo, and the residue remaining is taken up with dichloromethane, washed twice with water, dried over Na2SO4 and concentrated in vacuo. The oil remaining is purified by chromatography on Sephadex LH-20-100 (25 - 100p; mobile phase: methanol).

Yield: 65 mg (0.22 mmol; 94% of theory); colorless oil C1BH27NO4: calc.: C 64.5 H 9.1 found: C 64.6 H 9.2 13C-NMR spectrum (90.5 MHz; CDC13/TMS): = 169.88 (C-2); 157.51 (C-4); 85.42 (C-3); 70.40 (C-6); 66.22 (C-10); 60.44 (C-7); 55.36 (C-8); 43.56 (C-12); 38.48 (C-5); 37.15 (C-9); 31.35 (C-15); 28.18 (C-13); 26.61 (C-14); 22.44 (C-16); 21.10 (C-ll); 13.88 (C-17) Example 5: Preparation of 2,4-dioxo-6-(l,3-dioxo-but-l-yl)-oxy-10methyl-l-oxa-7-cyclodecene oxide (11.31) mg of the ten-ring lactone of the formula III are dissolved in 3 ml of absolute dichloromethane and 2 μΐ of absolute pyridine (10 mol-%) and 19 μΐ of diketene (1.05 equivalents) are added. The reaction components are stirred at room temperature for 24 h.

For working-up, the organic phase is diluted with 50 ml of dichloromethane, washed several times with water, dried over Na2SO4 and the solvent is removed in vacuo. The residue remaining is purified by means of flash chromatography on silica gel (mobile phase: CHCl3/hexane/methanol » 16x16:1).

Yield: 22.3 mg (0.75 mmol; 32% of theory); colorless oil C14H18O7 : calc ·: found: C 56.2 C 56.4 Η 6.0 H 6.1 •Ε 904375 - 31 Example 6: Glycosylation of the ten-ring lactone of the formula III by means of α-bromotetraacetylgalactose (11.16) mg of the ten-ring lactone of the formula III are 5 dissolved in 5 ml of CH2C12 and 170 mg of bromotetraacetylgalactose (2 equivalents), 150 μΐ of collidine (5 equivalents), 100 mg (about 2 equivalents) of AgC104 and 3 g of finely powdered molecular sieve (3A) are added with stirring at room temperature. The reaction mixture is stirred at room temperature for 2 days.

For working up, the reaction mixture is filtered, washed twice with water, dried over Na2SO4 and the solvent is removed in vacuo. The residue is purified by chromatography on silica gel (mobile phase: CHCl3/hexane/methanol = 8:8:1).

Yields 48 mg (0.88 mmol; 38% of theory); colorless oil (The reaction product is the orthoester according to spectroscopic data) ^2Α^32θ14 · CalC. · C 52.7 H 5.8 found: C 52.9 H 5.913C-NMR spectrum (90 .5 MHz; CDClg/TMS): 6 = 199.82 (C-4); 165.34 (C-2); 120.93 (C-12); 97.83 (C-14); 73.00 (C-15); 70.72 (C-16); 69.40 (C-6); 68.99 (C-10); 68.75 (C-18); 65.69 (C-17); 61.55 (C-19); 59.51 (C-7); 54.97 (C-8); 52.07 (C-3); 47.92 (C-5); 36.77 (C-9); 24.16 (C-13); 20.64 (C-ll); in addition, 3 acetyl groups with signals at 170.41; 169.96; 169.80; 20.69; 20.64 and 20.48 Example 7 Preparation of 3,3-dimethyl-2,4-dioxo-6-methoxy-10methyl-l-oxa-7-cyclodecene oxide (11.40) - 32 50 mg of the ten-ring lactone of the formula III are dissolved in 2 ml of absolute N,N-dimethylformamide and 0.5 ml of iodomethane and 400 mg of freshly precipitated silver oxide are added at room temperature. The reaction mixture is stirred at room temperature for 6 h.

For working-up, the reaction solution is filtered through filter aid. The solvent is then removed in an oil pump vacuum and the residue is purified by chromatography on silica gel (mobile phase: CHC13).

Yield: 55 mg (0.22 mmol; 92% of theory); colorless oil C33H2qO3: calc.: C 60.9 H 7.7 found: C 60.9 H 7.9 Example 8: Preparation of 2,4-dioxo-6-(4-fluoro-2-chlorophenyl)15 carbonyloxy-10-methyl-l-oxa-7-cyclodecene oxide (11.38) mg of the ten-ring lactone of the formula III are dissolved in 3 ml of absolute dichloromethane and 48 mg (1 equivalent) of dicyclohexylcarbodiimide, 41 mg (1 equivalent) of 4-fluoro-2-chlorobenzoic acid, and also 2 mg of Ν,Ν-dimethylaminopyridine are added. The reaction mixture is stirred at room temperature for 12 h.

For working-up, the mixture is filtered through filter aid, the organic phase is diluted with dichloromethane, extracted with water and dried over Na2SO4, and the solvent is removed in vacuo. The residue is purified by means of chromatography on silica gel (mobile phase: CHC13).

Yield: 52 mg (0.14 mmol; 60% of theory); colorless oil CnH16ClFO6: calc.: C 54.8 H 4.3 found: C 55.0 H 4.4 - 33 Example 9: Preparation of 2,4-dioxo-10-methyl-l-oxa-6-trimethylsilyloxy-7-cyclodecene oxide (II.9) mg of the ten-ring lactone of the formula III 5 (0.14 mmol) are dissolved in 5 ml of dichloromethane and 16.7 mg (1.1 equivalents) of trimethylchlorosilane and 14.4 mg (1.3 equivalents) of absolute pyridine are added. The reaction mixture is stirred at room temperature for 6 h.

For working-up, 50 ml of dichloromethane are added to the mixture, and the organic phase is washed twice with water, dried over Na2SO4 and the solvent is removed in vacuo. The reaction product is purified by means of flash chromatography on silica gel (mobile phase: CHC13).

Yield: 24.9 mg (0.87 mmol; 62% of theory) C13H22O5Si: calc.: C 54.7 H 7.6 found: C 54.5 H 7.7 Example 10: Preparation of 3-(bis-ethoxycarbonyl)-methylene-2,420 dioxo-6-hydroxy-10-methyl-l-oxa-7-cyclodecene oxide (11.17) mg of the ten-ring lactone of the formula III are dissolved in 3 ml of absolute chloroform and 0.1 ml (2.2 equivalents) of triethylamine and 60 μΐ (1.2 equivalents) of diethyl mesoxalate are added at a temperature of -20’C. After 2 h at -2O’C, the mixture is stirred at room temperature for 4 h.

For working-up, the mixture is freed from solvent and base in an oil pump vacuum and the pale yellow oil remaining is purified by means of chromatography on Sephadex LH-20 (mobile phase: methanol). - 34 Yield: 101.2 mg (0.27 mmol; 84% of theory); colorless oil C17H22Oa: calc.: C 55.3 H 5.8 found: C 55.1 H 6.0 Example 11: Preparation of 4,6-dihydroxy-10-methyl-l-oxa-2-oxo-7cyclodecene oxide (II.2) mg of the ten-ring lactone of the formula III (0.234 mmol) are dissolved in 5 ml of absolute tetrahydrofuran and 16.8 mg (1.15 mole equivalents) of sodium cyanoborohydride are added in portions at room temperature. The pH of the reaction solution is kept at pH 4 (indicator Methyl Orange) by addition of a solution of HCl/glacial acetic acid/THF. After 4 h, 50 ml of dichloromethane are added to the organic phase, the mixture is extracted twice with water and dried over Na2SO4, and the solvent is distilled off in vacuo. The residue remaining is purified by means of chromatography on silica gel (mobile phase: ethyl acetate).

Yield: 23.3 mg (0.11 mmol; 46% of theory) C10H16O5: calc.: C 55.3 H 7.4 found: C 55.5 H 7.5 Example 12: Preparation of 2,4-dioxo-10-methyl-l-oxa-6-tetrahydropyranyloxy-7-cyclodecene oxide (11.43) 50 mg of the ten-ring lactone of the formula III (0.23 mmol) are dissolved in 3 ml of absolute dichloromethane at room temperature and 29.5 mg (0.35 mmol; 1.5 mole equivalents) of dihydropyran and a spatula tip full of pyridinium p-toluenesulf onate (PPTS) are added.

The reaction mixture is allowed to stand at room temperature for 24 h. - 35 For working-up, the reaction mixture is washed twice with a little saturated ammonium chloride solution, dried over Na2SO4 and the solvent is removed in vacuo. The purification of the reaction product is carried out by means of flash chromatography on silica gel (mobile phase: CHCl3/hexane/methanol = 20:20:1).

Yield: 51.8 mg (17.4 mmol; 74% of theory) C15H22O6: calc.: C 60.6 H 7.5 found: C 60.4 H 7.4 (According to spectroscopic data, the reaction product is an approximately 1:1 mixture of the two diastereomeric tetrahydropyranyl ethers) Example 13: Preparation of 2,4-dioxo-10-methyl-l-oxa-5-cyclodec-7-ene oxide (1.1) mg of the ten-ring lactone of the formula III (0.23 mmol) are dissolved in 3 ml of absolute pyridine and 200 pi of p-toluenesulfonyl chloride are added. The mixture is then stirred at room temperature for 14 h.

For working-up, the mixture is poured onto 50 ml of dichloromethane, the organic phase is washed three times with water and dried over Na2SO4, and the solvent is removed in vacuo.

The reaction product is then dissolved in 2.5 ml of absolute toluene and stirred with 1 equivalent of diazabicycloundecene at 8O’C for 3 h. It is then again worked up as indicated above and the reaction product is purified by means of flash chromatography (silica gel; eluent CHC13) .

» Yield: 22 mg (0.11 mmol; 48% of theory); colorless oil Ο3θϊϊχ2θ4ι · calc · found C 60.9 H 6.2 C 61.2 H 6.2 Example 14: Preparation of 2,4-dioxo-10-methyl-l-oxa-7-cyclodecene 5 oxide (11.44) mg of the compound prepared in Example 13 are dissolved in 3 ml of tetrahydrofuran and about 4 mg of palladium on carbon (5% on active carbon) are added as a hydrogenation catalyst and the mixture is left in a hydrogen atmosphere with vigorous stirring for 1 h.

The catalyst is then filtered off through filter aid, the solvent is distilled off in vacuo and the remaining oil is purified by means of flash chromatography on silica gel (mobile phase: CHCl3/hexane/methanol = 16:16:1).

Yield: 17 mg (0.086 mmol; 92% of theory) C10H14|O4: calc.: C 60.3 H 7.0 found: C 60.6 H 7.1 Table 1 below gives a survey of the compounds prepared and of those which were prepared analogously to Examples 1-14. fM 5S1 zo o vt x-^ xz O ''-J **** o s PM ι O O PM cn IM o Χ·Ζ I z-b fM 8 8 8 s 8 o o o o x»-z X·^ » z o O Z-^ X σι X o 1 X CJ X o cn II o CJ fM o X a PM UJ ι o x UJ II o II o II o II a IM O σι \z o X UJ X o X o&o uj o fM O » O X PM *< -J PM ι o fM im σι cn cn I O X UJ ll II o o ιι ο A O X o PM > PM fM UJ oso Ό ΓΜ* *< -4 0*-* X X o cn im x o o UJ • PM O Ό «< PM 0=50 PM o << zr X & r* IT fe *1 PM PM 50 PM nj fM »30-4 & Qi -*» x-z o> 00 σι 00 cn 4b σι cn *M 4b 00 cn PM PM 1— PM cn σι o 4b σι X X X X X X X X X X X *xj cn X - LZ Yield βε h/RT c) 4 94 % Continuation of Table 1 6E Cl II.38 O-C -?oYf =0 Η H 4-F-2-Cl-benzoic acid, DOC 12 h/RT a) 8 60 X 44 »4 44 44 44 44 44 44 44 1—4 » • • • • 44 — • » • CU cu cu cu bU • CU cu CU ru **4 at cn cu no no 44 o to a I θ © I o g z a 8 o o I o 1 oan z 0X0 o ai> ru □so os© o X 1¼ t X 1 X C7 o cn o z nu IT 1 {Γ o z z 1 II . X 1 IM 1 σ J ru r> 1 o σ» X o»o ru ru TT cn z Ut * X O z * II o 4b Z X Tl 1 z 1 ί w o -n o 44^ o nu o o 1 X nu O cu o at X nu X Cu 1 bU O k—X at X Jb It u II II II II 1 u II & II o o o © o o 8 o o σ X o «» cu z X z ru z z χ o z z X o ru at X X cn cn o X nu X X x o X z z o z z z at ru X X cn cn 00 00 bU ςη ω to at -4J >4 10 00 σ» cu * 'J at at o CU fM 00 * tn nu 00 σ M X X 4 4 4 H 4 4 S’ Continuation of Table 1 - Ofr IE 904375 * r» §5* - 42 Inhibition of cholesterol biosynthesis The inhibition of cholesterol biosynthesis by the compounds according to the invention was detected in cell culture experiments using HEP-G2 cells by means of the effect on the incorporation of l*C-sodium acetate Into cholesterol. For this purpose, monolayers of HEP-G2 cells were preincubated for 1 hour in lipoprotein-free nutrient medium with appropriate concentrations of the compounds according to the invention; after addition of the 14C-labeled precursor “c-sodium acetate, the incubation was continued for 3 hours. One part of the cells was then subjected to alkaline hydrolysis, after previous addition of a 3H-cholesterol internal standard. The lipids of the hydrolyzed cells were extracted with chloroform-methanol.

This lipid mixture was separated by preparative thinlayer chromatography after addition of carrier cholesterol, the cholesterol band was isolated after visualization and the amount of 14C-cholesterol formed from the 14C-precursor was determined scintigraphically.

Cell protein was determined in an aliquot part of the cells so that the amount of 14C-cholesterol formed from 14C-precursor in unit time per mg of cell protein could be calculated. The control was used as a comparison of the inhibitory action of an added compound according to the invention so that the inhibition of cholesterol biosynthesis at a certain molar concentration of the compound according to the invention in the medium could be indicated directly. To evaluate the potency of the compounds according to the invention, the inhibitory action of lovastatin was determined in the same experimental batch. The intactness of the cell culture was determined in aliquot portions of the cell culture and the lack of cell damage by the action of the compound according to the invention was evaluated morphologically (light microscope) and biochemically by determination of the release of lactate dehydrogenase in the incubation medium.

The result of this investigation is represented in Table 2. The action of the substances according to the invention as cholesterol biosynthesis inhibitors can be clearly recognized. In the concentrations tested, the compounds according to the invention had no cytotoxic effect in a period of 4 hours.

The inhibition of hepatic cholesterol biosynthesis has effects on the reduction of serum lipids, as it was possible to detect in the chronic experiment on the male rat (Table 2). l-l HI HI HI Hl HI H H Hl H HI HI HI HI • HI HI HI HI HI • • w • H* • • • • • M M • LO M GJ GJ GJ 4k Ul M 4k O -J σι Ul 4k Ul Ul X X m ui 01 ui XX* X °l °, β» -4 · OB Λ °i O ’ £ § £ § Ul to 4k 4k -J gj O UJ vo σι oo 00 H· (a* KO KO Ul KO O Ul 4k σι GJ σι σι gj X X H· I-* o o I I (O Inhibition of cholesterol biosynthesis in HEP-G2 cell culture MMMOlOlMOll-MM ·········· MhJGJUlUlGJUlGJMWOlOlOlWOl XXXXXXXXXXXXXXX ,-,|_,|_.|_.|_.|_.|_.|_.(-·|_.|_>Η·Η·Η·Ι— O O Q p p p p p p p p p p p p οοα,σ.ο,(»θΒαΒθΒσ,θΒ(>σ.ώί(>αι w HI 4k GJ 9) H oo SJ Ul X M Ul X M Ul X o p p o p p p p p p o H* GJ004k-J'JO00GJ~JOH· CB 0 i=! , Ϊ £· of Table 2 - 5» IE 904375 - 46 Experimental procedure Daily in the morning, groups of male rats of the strain HOE: WISKf (SPF 71) having a starting weight of about 240 g received the compounds according to the invention in polyethylene glycol 400 by stomach tube, the respective control group receiving only the vehicle. 24 hours after the last (7th) administration and after withdrawal of food for 24 hours, blood was taken and the lipoproteins in the serum obtained were separated from the pool of one rat group by means of the preparative ultracentrifuge technique. In this technique, the following density limits for the separation of VLDL, LDL and HDL were used: VLDL 1.006 15 LDL 1.04 HDL 1.21 VLDL = very low density lipoprotein LDL = low density lipoprotein HDL = high density lipoprotein Fully enzymatic methods by Boehringer/Mannheim were used for the determination of cholesterol and triglycerides, the method of Lowry et al. J. Biol. Chem., 193, 265 (1951) was used for the determination of protein. As is shown by Example II.3 (see Table 3), 7-day's oral admini25 strat ion led in a dose lower by one third than the standard preparation clofibrate to a clear reduction of LDL and VLDL, accompanied by an increase in the HDL fraction. An increase in the vasoprotective index (HDL cholesterol s LDL cholesterol) resulted from this with an about 8-fold higher potency than clofibrate. The use of the substance was tolerated by the rats without difficulty. In contrast to clofibrate, no hepatomegaly occurred.

Ul tb Ol I— I + GJ NJ Ol NJ GJ GJ 4k • · · NJ 00 00 4k «U 00 It + + ί- ο N— Ul o KO H· <*» <* dP I I NJ GJ (D u £· £ 8· Influence of serum lipoproteins in Vasoprotective Index VLDL - Lt IE 904375

Claims (13)

1. Patent claims:

1. A 10-ring lactone of the formula I or II R 1 5 in which A 1 and A 2 are identical or different and are H or Ci-C*alkyl, the C^-C^-alkyl radical being unsubstituted, or monosubstituted or polysubstituted by OH or phenyl, the phenyl radical in turn being unsub10 stituted, or monosubstituted or polysubstituted by halogen, CF 3 or Cx-Cj-alkyl or in which A 1 and A 2 together form a radical of the formula in which A 3 and A 3 ' are identical or different and are Cj-Ca-alkoxycarbonyl, phenyl or C^-C^-alkyl, or in which A 1 and A 2 are together a radical of the formula )H or in which A* is phenyl or Cj-Cg-alkyl, R 1 is -H, =0 or -OR 3 and R 2 is =0, -OR*, =N-R 5 or ,5' , where in the case of the formula II for R 2 = =N-R 5 and A 1 = H, this can also be present as the compound of the formula II’ c (II') ?Η, 0 in which R 3 is H, Ci-Ce-alkyl, benzyl, allyl, MEM, MOM, SEM, THP, 4'-methoxytetrahydropyran-4'-yl, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, dimethylcyclohexylsilyl, dimethyl tertiary-butylsilyl or a sulfonic acid ester of the formula -SO 2 R 7 in which R 7 is Cx-Cio-alkyl, phenyl or p-methylphenyl or R 3 is a mono- or disaccharide whose OH groups are unprotected or are protected by protecting groups or X X , 11 8 J 8 R 3 is a radical of the formula -C-Z-R or -C-Z-(CH 2 ) n -R in which X is 0 or S Z is 0, -N-R 8 , -N-(CH 2 ) n -R 8 or -NH n is 1 to 3 and R 8 is Cx-Ce-alkyl, C 3 -C e -cycloalkyl, aryl, pyridyl, 20 pyrimidyl or pyrazinyl or Ϊ . R 3 is a radical of the formula -C-R in which 25 R 9 is Ci-Cao-alkyl, C 2 -C 16 -alkenyl having 1-3 double bonds, C 2 -C 16 -alkynyl having 1-3 triple bonds, C 3 -C B -cycloalkyl, aryl or heteroaryl, the alkyl, alkenyl, alkynyl and cycloalkyl radicals being unsubstituted, or monosubstituted or poly30 substituted by COOA 5 , halogen, aryl, aryloxy, heteroaryl, heteroaryloxy, NHCOCH 3 , COA 5 or SO 3 A 5 in which A 5 is Cx-C^-alkyl or H and in which all said aryl, heteroaryl, pyridyl, pyrimidyl and - 50 pyrazinyl radicals are unsubstituted, or monosubstituted or polysubstituted by halogen, nitro, cyano, hydroxyl, trifluoromethyl, amino, carboxyl, Οχ-C^-alkylcarbonyl, C x -C 4 -alkoxy5 carbonyl, Cx-Cjo-alkyl, C x -C 4 -alkoxy or NHCOCH 3 and in which R 4 , independently of R 3 , has the meanings indicated above for R 3 and R 5 and R 5 ' are identical or different and are hydrogen, 10 Cx-Cxo-alkyl, C 2 -C 10 -alkenyl, benzyl, phenyl or phenylethyl, said radicals being unsubstituted or substituted by hydroxyl, halogen, NH 2 or OCH 3 , and the physiologically tolerable salts, excluding the compound of the formula I in which R 2 is 15 -OR 4 where R 4 = H and also excluding the compound of the formula II in which R 1 is -OR 3 and R 2 is =0 where R 3 = H.

2. A 10-ring lactone of the formula I or II as claimed in claim 1, in which A 1 and A 2 are identical or different and are H or Cx-C 2 alkyl, the Cx-Ca-alkyl radical being unsubstituted or monosubstituted by phenyl, the phenyl radical in turn being unsubstituted or monosubstituted by halogen, CF 3 or C x -C 3 -alkyl or in which A 1 and A 2 together are a radical of the formula in which A 3 and A 3 * are identical or different and are Cx-Cj-alkoxycarbonyl or phenyl or in which A 1 and A 2 together are a radical of the formula in which A 4 is phenyl, R 1 is -H, =0 or -OR 3 and -K R 2 is =0, -OR 4 , “N-R 5 or , where in the case of the formula II for R 2 = =N-R 5 and A 1 = H, this can also be present as the compound of the formula II', as indicated in claim 1, in which 5 R 3 is H, Ci-Cg-alkyl, benzyl, allyl, MEM, MOM, SEM, THP, 4'-methoxytetrahydropyran-4'-yl, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, dimethylcyclohexylsilyl, dimethyl tertiary-butylsilyl or a sulfonic acid ester of the formula -SO 2 R 7 in which R 7 10 is methyl, phenyl or p-methylphenyl or R 3 is a pyranosyl radical whose OH groups are unprotected or protected by protecting groups or 15 XX 3. 11 β 8 R 3 is a radical of the formula -C-Z-R or -C-Z-(CH 2 ) n -R in which X is 0, I β I B I Z is 0, -N-R 8 , -N-(CH 2 ) n -R 8 or -NH 20 n is 1 to 3 and R 8 is Ci-Ce-alkyl, C 3 -C 6 -cycloalkyl or aryl or R 3 is a radical of the formula -C-R® 25 in which R® is C^C^-alkyl, C 2 -C 16 -alkenyl, C 3 -C 6 -alkynyl, C 3 -C 6 -cycloalkyl, aryl or heteroaryl, the alkyl, alkenyl, alkynyl and cycloalkyl radicals being unsubstituted, or monosubstituted or poly30 substituted by COOA 5 , halogen, phenyl, phenyloxy, NHCOCH 3 , COA 5 or SOaA 5 A 5 being C^C^alkyl or H and in which all said aryl and heteroaryl radicals are unsubstituted, 35 or monosubstituted or polysubstituted by halogen, cyano, trifluoromethyl, carboxyl, 0 χ -0 4 alkylcarbonyl, C x -C 4 -alkoxycarbonyl, C x -C 7 -alkyl, C x -C 4 -alkoxy or NHCOCH 3 - 52 and in which R 4 , independently of R 3 , has the meanings indicated above for R 3 and R 5 and R 5 ’ are identical or different and are hydrogen, 5 Cx-Cxo-alkyl, C 2 -C 10 -alkenyl, benzyl, phenyl or phenylethyl, said radicals being unsubstituted or substituted by hydroxyl, halogen, NH 2 or OCH 3 , and the physiologically tolerable salts, excluding the compound of the formula I in which R 2 is 10 -OR 4 where R 4 = H and also excluding the compound of the formula II in which R 1 is -OR 3 and R 2 is =0 where R 3 = H.

3. A 10-ring lactone of the formula I or II as claimed in claim 1 or 2, in which A 1 and A 2 are identical or different and are H or methyl, 15 the methyl radical being unsubstituted or monosubstituted by phenyl, or in which I A 3 A 1 and A 2 together are a radical of the formula =/ in which A 3 and A 3 ' are identical and are ethoxycarbonyl R 1 is -H, =0 or -OR 3 and r 5 R 2 is =0, -OR 4 , =N-R 5 or -Nft , , where in the case of X R 5 25 the formula II for R 2 = =N-R 5 and A 1 = Η , this can also be present as the compound of the formula II', as indicated in claim 1, in which R 3 is H, methyl, benzyl, THP, trimethylsilyl, dimethyl 30 tertiary-butylsilyl or a sulfonic acid ester of the formula -SO 2 R 7 in which R 7 is p-methylphenyl or R 3 is a glucosyl, galactosyl or lactosyl radical whose 35 OH groups are unprotected or are protected by acetyl groups or benzyl groups, these said sugar radicals - 53 either being glycosidically bonded or linked via an orthoester or X X 5 R 3 is a radical of the formula -C-Z-R or -C-Z-(CH 2 ) n -R in which X is O, . I Z is -NH n is 1 and 10 R 8 is C 4 -C 8 -alkyl, or aryl or , 11 β R is a radical of the formula -C-R in which 15 R 9 is Cx-C^-alkyl, C 2 -C 8 -alkenyl, C 3 -C 6 -cycloalkyl, phenyl, thienyl or furyl, the alkyl, alkenyl, and cycloalkyl radicals being unsubstituted, or monosubstituted or polysubstituted by COOA 5 , F, Cl, Br, phenyl, phenyloxy, NHCOCH 3 , or COA 5 , A 5 20 being methyl or H and in which all said aryl or phenyl radicals are unsubstituted, or monosubstituted or polysubstituted by F, Cl, Br, trifluoromethyl, carboxyl, Ci-C^-alkyl25 carbonyl, C x -C 4 -alkoxycarbonyl, Ci-Cj-alkyl, methoxy or NHCOCH 3 and in which r* , independently of R 3 , has the meanings indicated above for R 3 and 30 R 5 and R 5 ’ are identical or different and are hydrogen, Ci-Cg-alkyl, benzyl, phenyl or phenylethyl, said radicals being unsubstituted or substituted by F, Cl, Br or OCH 3 , and the physiologically tolerable salts, 35 excluding the compound of the formula I in which R 2 is -OR* where R* - H and also excluding the compound of the formula- II in which R 1 is -OR 3 and R 2 is =0 where R 3 = H.

4. A process for the preparation of the 10-ring lactone - 54 according to formula I or II, as claimed in claim 1, wherein a) the compound of the formula III 5. Is reacted with a compound of the formula IV /X* (IV) in which Q is OH, chloride, bromide, imidazolide or an acid anhydride and 10 R® has the meanings mentioned in claim 1 for formula I and II, or wherein b) the compound of the formula III is reacted with a compound selected from amongst: 15 dihydropyran, 4-methoxy-5,6-dihydropyran, Cj-Cealkyl, benzyl or allyl chloride, bromide or iodide or MEM, MOM or SEM chloride, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, dimethylcyclohexylsilyl or dimethyl tertiary-butylsilyl chloride, 20 sulfonic acid halides of the formula Hal-SO 2 R 7 in which Hal is chlorine, bromine or iodine and R 7 has the abovementioned meanings, isocyanates of the formula 0 = C = N-R 8 or 25 0 = C = N-(CH z ) n -R 8 , isothiocyanates of the formula S = C = N-R 8 or - 55 10 S = C = N-(CH 2 ) n -R 8 , carbamoyl halides of the formula Hal '-CO-N-(R 8 ) 2 or Hal'-C0-N-[ (CH 2 ) n -R 8 ]2 thiocarbamoyl halides of the formula Hal' -CS-N- (R 8 )2 or Hal'-CS-N-[ (CH 2 ) n -R B ]2 or compounds of the formulae Hal '-C(O)-O-R 8 , Hal'-CO-(CH2) n -R 8 , Hal'-CS-O-R 8 or Hal'-CS-O-(CH 2 ) n -R 8 in which n and R 8 have the meanings mentioned in claim 1 in formula I and II and Hal' is chlorine or bromine, or wherein c) the compound of the formula III is reacted with a compound of the formula V j>5 X R H-N ^R 5 ’ (V) 15 in which R 5 and R 5 ’ are defined as indicated in claim 1 in formula I and II, and the resulting C-C double bond in the compound of the formula II' is optionally hydrogenated, or wherein 20 d) the compound of the formula III is reacted with a compound of the formula VI R 3 - Hal (VI) in which R 3 is a mono- or disaccharide and Hal is as defined above, or wherein e) the compound of the formula III is oxidized to a compound of the formula VII (VII) is reduced to a or wherein f) the compound of the formula III - 56 compound of the formula or wherein g) a compound of the formula III is subjected to elimination to give the enone and the C-C double bond is optionally hydrogenated, a compound of the formula (Xi: being formed, or wherein h) the compound of the formula III is reacted with a compound of the formula VIII and/or VIII' in which A 1 and A 2 are formula I and II except for the meaning A 1 - Abg A 2 - Abg as defined (VIII) (VIII') in claim 1 in and Abg is - 57 chlorine, bromine, iodine, sulfate, mesylate or tosylate or wherein i) the compound of the formula III is reacted with a compound of the formula IX (IX) in which A 3 and A 3 ' are as defined in claim 1 in formula I and II, or wherein 10 j) the compound of the formula III is reacted with a compound of the formula X in which A* is as defined in claim 1 in formula I and II or wherein k) the compound of the formula III is reacted with diketene a compound of the formula II being formed where R 1 = 0-C(0)-CH 2 -C(0)-CH 3 or wherein 1) a compound of the formula III is reacted with a mesoxalic acid diester, a compound of the formula II being formed where A 1 and A 2 in which A 3 and A 3 ’ are as defined in claim 1 in formula I and II, or wherein two or more of the abovementioned process variants a-1 are carried out successively and wherein the compounds I or II are then optionally converted into their physiologically tolerable salts.

5. A 10-ring lactone of the formula I or II as claimed in one of claims 1 to 3, but including the compound of the formula I in which R 2 is -OR* where R* = H, and 5 including the compound of the formula II in which R 1 is -OR 3 and R 2 is =0, where R 3 = H, for the preparation of a pharmaceutical having cholesterol biosynthesis-inhibiting action.

6. A compound of the formula I or II as claimed in one 10 of claims 1 to 3 for use as a pharmaceutical.

7. A compound of the formula I or II as claimed in one of claims 1 to 3 for use as a pharmaceutical having hypolipidemic action.

8. A pharmaceutical containing at least one compound of 15 the formula I and/or II as claimed in one of claims 1 to 3.

9. A process for the production of a pharmaceutical, wherein at least one compound of the formula I and/or II as claimed in one of claims 1 to 3 is incorporated into 20 the pharmaceutical.

10. A compound as claimed in claim 1, substantially as hereinbefore described and exemplified.

11. A process for the preparation of a compound as claimed in claim 1, substantially as hereinbefore described and exemplified.

12. A compound as claimed in claim 1, whenever prepared by a process claimed in claim 4 or 11.

13. A pharmaceutical as claimed in claim 8, substantially as hereinbefore described.