AU657263B2 - HMG-CoA reductive inhibitors - Google Patents

Description

WO 91/15482 PCT/GB91/00357 1 1 HMG-CO A REDUCTIVE INHIBITORS 2 3 This invention relates to pharmaceutically active 4 compounds, which are substituted decalins. The compounds of the present invention are inhibitors of 6 the enzyme 3-hydroxy-3-methylglutaryl coenzyme A 7 reductase (HMG-CoA reductase), the rate limiting enzyme 8 in the biosynthesis of cholesterol in mammals including 9 man, and as such are useful in the treatment of hypercholesterolaemia and hyperlipidaemia. Clinical 11 evidence shows that reduction of serum cholesterol 12 levels leads to a decreased risk of heart disease.

13 14 The natural fermentation products compactin (disclosed by A. Endo, et al. in Journal of Antibiotics, 29, 16 1346-1348 (1976)) and mevinolin (disclosed by A.W.

17 Alberts, et al. in J. Proc. Natl. Acad. Sci. U.S.A., 18 77 3957 (1980)) are very active 19 antihypercholesterolaemic agents which limit cholesterol biosynthesis by inhibiting the enzyme 21 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) 22 reductase, the rate-limiting enzyme and natural point 23 of cholesterolgenesis regulation in mammals, including 24 man. Compactin (R H, a double bond) and mevinolin (R a-CH 3 a double bond; also known as lovastatin) 26 have the structures shown below:- 27 28 HO 0 29 0 31 32 33 WO 91/15482 PCT/GB91/00357 2 1 Also known in the art are the natural products 2 dihydrocompactin (R H, a single bond) disclosed by 3 Y.K.T. Lam et al., Journal of Antibiotics, 34, 614-616 4 (1981), dihydromevinolin (R a-CH 3 a single bond) disclosed by G. Albers-Schonberg et al., Journal of 6 Antibiotics, 34, 507-512 (1981), and eptastatin (R 7 8-OH, a double bond) disclosed by N. Serizawa et al., 8 in Journal of Antibiotics, 36, 604-607 (1983;.

9 US-A-4293496 (Willard) discloses a number of 11 semi-synthetic analogues of mevinolin having the 12 structure 13 110 0 14 0 16 17

R

18 19 21 22 23 where the dotted lines represent single or double bonds 24 and R is C 1 8 straight chain alkyl, C3_10 branched chain alkyl except (S)-2-butyl, C 3 -1 0 cycloalkyl, C2- 10 26 alkenyl, C,- 10

CF

3 substituted alkyl, halophenyl, 27 phenyl C1- 3 alkyl and substituted phenyl C1-3 alkyl.

28 29 US-A-4444784, US-A-4661483, US-A-4668699, and US-A-4771071 (Hoffman) disclose compounds of similar 31 structure where the R group contains extra functional 32 groups, for example ether, amide and ester groups.

33 WO 91/15482 PC'/G B91/00357 3 In J. Med. Chem., 29, 849-852 (1986), W.F. Hoffman et al. report the synthesis and testing of a number of the analogues referred to above, the preferred compound (now known as simvastatin) having the structure HO 0O US-A-4820865 (Terahara) discloses compounds having the structure 0 R O wherein R represents a C_- 10 alkyl group.

In general, the above patents also cover compounds in which the delta lactone has been hydrolysed to a delta hydroxy acid or a salt of that acid.

WO 91/15482 W CT/1 9)1/00357 4 1 None of the cited patents and articles disclose or 2 suggest the possibility of preparing the compounds of 3 the present invention. The unique pattern of 4 substituents on the decalin ring system differs from the cited art, whilst the compounds exhibit potent 6 HMG-CoA activity.

7 8 The present invention provides novel decalin based 9 compounds which are potent inhibitors of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) 11 reductase and, therefore, are useful in the treatment 12 or prevention of hypercholesterolaemia, 13 hyperlipoproteinaemia and atherosclerosis.

14 According to a first aspect of the invention there is 16 provided a compound of either formulae I and II: 17 18 HO HO R 19

Q

0 21 b bj 22 R'CO.O R'CO.O 23 R s

R

S

24

(II)

25 HO a HO 26 27 wherein: .28 29 R 1 represents C_-8 alkyl, C 3 -8 cycloalkyl, C 3 _g cycloalkyl(Cl_ 8 )alkyl, 31 32 C 2 8 alkenyl, or C 1 -6 alkyl substituted phenyl group; 33 NO 91/15482 PCT/GB91/00357 Macb 1992 s 2D 03 92 1 R 3 represents a hydrogen atom or a substituent R 4 or M; 2 3 R 4 represents a C1_ 5 alkyl group, or a C 1 -5 alkyl group 4 substituted with a group chosen from substituted phenyl, dimethylamino, or acetylamino; 6 7 R 5 represents a hydrogen atom or a C, 1 3 alkyl group; 8 9 M represents a cation capable of forming a pharmaceutically acceptable salt; 11 12 Q represents C=O or CHOH; and 13 14 oe-ea-e4 a)and b s5 -indpendentl-y- a single or double bond.

16 17 The term "C 1 -8 alkyl" refers to a straight or branched 18 chain alkyl moiety having one to eight carbon atoms, 19 including for example, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, pentyl, dimethyl-propyl, 21 hexyl, and octyl, and cognate terms (such as "C1-8 22 alkoxy") are to be construed accordingly.

23 24 The term "C 3 8 cycloalkyl" refers to a saturated alicyclic moiety having from 3 to 8 carbon atoms 26 arranged in a ring and includes, for example, 27 cyclopropyl, cyclobutyl, cyclopentyl, and cyclooctyl.

28 29 The term "C2_ 8 alkenyl" refers to a straight or branched chain alkyl moiety having two to eight carbon 31 atoms and having in addition at least one double bond, 32 33 iPCT In, oa! pplication United Kerdomnr nt Office SUSTITUTE SHEET I PCT inic.:ia;ona! Application y P.-r 1 i 1r*I>i 1 rO 91/15482 PCT/GB91/00357 6 1 of either E or Z stereochemistry where applicable. This 2 term would include, for example, vinyl, l-propenyl, 1- 3 and 2-butenyl and 2-methyl-2-propenyl.

4 The term "substituted", as applied to a phenyl or other 6 aromatic ring, means substituted with up to four 7 substituents each of which independently may be C1-6 8 alkyl, C 1 -6 alkoxy, hydroxy, thiol, amino, halo 9 (including fluoro, chloro, bromo, and iodo), trifluoromethyl or nitro.

11 12 The phrase "a pharmaceutically acceptable salt" as used 13 herein and in the claims is intended to include 14 non-toxic alkali metal salts such as sodium, potassium, calcium and magnesium, the ammonium salt and salts with 16 non-toxic amines such as trialkylamines, dibenzylamine, 17 pyridine, N-methylmorpholine, N-methylpiperidine and 18 other amines which have been used to form salts of 19 carboxylic acids.

21 There are several chiral centres in the compounds 22 according to the invention because of the presence of 23 asymmetric carbon atoms. The presence of several 24 asymmetric carbon atoms gives rise to a number of diastere'aisomers with the appropriate R or S designated 26 stereochemistry at each asymmetric centre. General 27 Formulae I and II and, where appropriate, all other 28 formulae in this specification are to be understood to 29 include all such stereoisomers and mixtures (for example racemic mixtures) thereof.

31 32 WO 91/15482 PCT/GB91/00357 7 1 Disregarding any asymmetric centers that may be present 2 in the groups R 1

R

3 and R 5 the preferred relative 3 and absolute stereochemistry is as shown in formula 4 III. More specifically for the compound III the Cahn, Ingold, Prelog designations for the absolute 6 configurations are 4a(R), 8(S), 7 8a(S), 8 9 11 o 12 6' 13 14 11 8 16 6 4a 2 17 HO 18 (III) 19 21 It is preferred that all of the compounds of Formulae I 22 and II should have (wherever possible) the same spacial 23 orientation of groups at each chiral carbon atom and 24 therefore belong to the same stereochemical series. The R-S designation for each center may not be identical to 26 that found for compound III because of the details of 27 the sequence rules for determining that designation.

28 Clearly in compounds of Formula II in which Q is the 29 group C=0 then the carbon atom labelled C-6' is not an asymmetric centre.

31 32 33 WO 91/15482 PCT/GB91/00357 8 1 In compounds of Formula II in which Q is the group 2 CHOH, the preferred stereochemistry is that in which 3 the two carbon atoms bearing the hydroxy groups have 4 the same spatial arrangement as the corresponding carbon atoms in the lactone in compound III. The 6 preferred isomer is referrred to as the syn diol.

7 8 The preferred compounds include those in which 9 independently or in any combination: 11 R 1 represents C 4 -6 branched alkyl; 12 R 3 is R 4 13 R 4 represents C1-5 alkyl and more preferably methyl or 14 ethyl;

R

5 represents methyl; 16 Q represents CHOH; and/or 17 b is a single bond.

18 19 Each M is preferably free from centres of asymmetry and is more preferably sodium, potassium or ammonium, and 21 most preferably sodium. For simplicity, each formula in 22 which an M appears has been written as if M were 23 monovalent and, preferably, it is. However, M may also 24 be divalent or trivalent and, when it is, it balances the charge of two or three carboxylic acid groups, 26 respectively. Thus Formula II and every other formula 27 containing an M embraces compounds wherein M is 28 divalent or trivalent, e.g. compounds containing two or 29 three mono carboxylate-containing anions per cation M.

31 Particularly preferred compounds are 32 33 WO 91/15482 PCT/GB91/00357 9 1 1) Methyl 2S, 4aR, 6R, 8S, 8aS)-6-hydroxy- 2 8-[(2(S)-methyl-l-oxobutyl)oxy]-l, 2, 4a, 5, 6, 7, 3 8, 8a-octahydro-2-methylnaphthalenyl]}-3(R),5(R)- 4 dihydroxyheptanoate 6 2) (IS, 2S, 4aR, 6R, 8S, 8aS, 4'R, 6'R, 7 2, 4a, 5, 6, 7, 8, 8a-octahydro-6-hydroxy- 8 2-methyl-8-[(2"-methyl-l"-oxobutyl)oxy]-1-naphtha- 9 lenyl]ethyl] tetrahydro-4'-hydroxy-2H-pyran-2'-one 11 3) Sodium 2S, 4aR, 6R, 8S, 8aS)-6- 12 hydroxy-8-[(2(S)-methyl-l-oxobutyl)oxy]-1, 2, 4a, 13 5, 6, 7, 8, 8a-octahydro-2-methylnaphthalenyl])- 14 16 For simplicity the compounds of Formula II may be 17 subdivided according to the exact form of R 3 and Q.

18 Thus the compounds in which Q is the group C=O and R 3 19 is a group of formula R 4 are considered to be compounds of the subgroup IIa, whereas if R 3 is M the ketones are 21 in the subgroup lie. Compounds in which Q is the group 22 CHOH and R 3 is a group of form R4 make up the subgroup 23 IIb, when R 3 is hydrogen the compounds are of subgroup 24 IIc, and when R 3 is a group of formula M the compounds are of the subgroup IId.

26 27 The present invention also provides novel processes for 28 the preparation of the compounds of general formulae I 2'9 and II as well as certain intermediates in their preparation, as will now be described by reference to 31 the drawings, in which:- 32 33 WO 91/15482 PCT/GB91/00357 Figure 1 Figure 2 shows reaction scheme I, which shows the interconversion of compounds of general formula I with subgroups IIa, IIb, IIc and IId; shows reaction scheme II, which shows a preparative route of compounds of subgroups IIa and IIe from compounds of general formula XIV, which in turn are preparable from compounds of general formula VII; shows reaction scheme III, which shows a different preparative route of compounds of general formula XIV, this time from compounds of general formula XV; and shows reaction scheme IV, which shows a preparative route of compounds of general formulae VII and XV from compounds of general formulae XXII, which in turn may be prepared from compounds of general formula XXIII.

Figure 3 Figure 4 The compounds of the various subgroups IIa-IId of general formula II (hereafter referred to as general formulae IIa to IId), and those of general formula I, may be prepared by the general reaction route shown in Scheme I in which R 1

R

4

R

5 and M are as previously defined. Unless the context otherwise requires, substituents in the general formulae in the Schemes I and II have the same values as the corresponding substituents in general formulae I and II.

WO 91/15482 PCI'/G1191/00357 11 1 According to a second aspect of the invention, there is 2 provided a process for the preparation of a compound of 3 either of general formulae I and II, the process 4 comprising: 6 deprotecting and optionally reducing a compound of 7 general formula XIV as shown in Scheme II in Figure 2 8 to form a compound of general formula IIa; and 9 optionally after step converting a compound of 11 general formula I or IIa directly or indirectly into 12 another compound of general formula I or II.

13 14 A etone of general formula IIa may be reduced to a dihydroxy ester of general formula IIb by reduction of 16 the ketone group with a reducing agent such as those 17 well known in the art e.g. sodium borohydride, sodium 18 cyanoborohydride, zinc borohydride, lithium 19 tri-s-butylborohydride or other similar reducing agents that will not reduce the ester functionality.

21 Preferably, the reaction is carried out in such a 22 manner as to maximize the production of the preferred 23 syn isomer of the coimpound of general formula IIb. The 24 stereoselective reduction of compounds of general formula IIa is preferably carried out in two stages, in 26 the first stage the ketone ester is reacted with a 27 trialkylborane, preferably tri-n-butyl borane, or an 28 alkoxydialkylborane, preferably methoxydiethylborane or 29 ethoxydiethylborane (Chemistry Letters, 1987, 1923-1926) at a temperature between -78°C and ambient 31 temperature in an inert organic solvent such as 32 tetrahydrofuran, diethyl ether, or 1,2-dimethoxyethane, 33 and optionally in the presence of a protic solvent such WO 91/15482 PCT/GB91/00357 12 1 as methanol or ethanol, and preferably in a mixture of 2 tetrahydrofuran and methanol. The complex which is thus 3 produced is then reduced with sodium borohydride at a 4 temperature between -78*C and -20"C. The resulting compound of general formula IIb produced from the 6 stereoselective reduction contains two asymmetric 7 carbon atoms bearing hydroxyl groups in a syn 8 configuration. Thus reduction of the ketone radical 9 under the preferred conditions described herein produces mostly the syn isomers of compounds of general 11 formula lib and only a small amount of the less 12 preferred anti isomers.

13 14 The ratio of isomers produced will vary according to the specific compound utilized and the reaction 16 conditions employed. Normally, this ratio will be 17 approximately 9:1 to 9.8 0.2. However, the use of a 18 non-specific reduction method will normally produce a 19 near 1:1 mixture of diastereoisomers. Nevertheless, the mixture of isomers may be separated and purified by 21 conventional techniques and then converted to the 22 compounds of general formula I in a conventional manner 23 well-known to those skilled in the art.

24 The compounds of general formula IIb may be cyclised to 26 the corresponding lactones of general formula I for 27 example by heating in an inert organic solvent such as 28 benzene, toluene or xylene and azetropically removing 29 the alcohol which is produced. Preferably, the lactonisation is carried out by heating the compounds 31 of general formula IIb with an acid, preferably 32 E-toluenesulphonic acid, in benzene or toluene, 33 evaporating the solvent and alcohol thus formed, and WO 91/15482 PCT/GB91/00357 8 13 1 repeating the process until all of the compound of 2 general formula IIb has been consumed. If the relative 3 stereochemical configuration of the two carbon atoms 4 bearing the hydroxy groups are established as syn in general formula lid, then lactonisation will produce 6 the preferred trans lactone of general formula I, 7 otherwise the lactonisation will produce a mixture of S trans and cis lactones.

9 A compound of general formula IId may be prepared from 11 a compound of general formula IIb or a compound of 12 general formula I by hydrolysis, preferably hydrolysis 13 with a base such as lithium hydroxide, sodium hydroxide 14 or potassium hydroxide in a mixture of water and an organic solvent such as methanol, ethanol or 16 tetrahydrofuran at a temperature between O0C and 17 inclusive, preferably at ambient temperature. The 18 cation in compounds of general formula IId is usually 19 determined by the cation of the hydroxide employed; however, the cation may then be exchanged for another 21 cation for example by treatment with an ion-exchange 22 resin.

23 24 The compounds of general formula IIc may be obtained from compounds of general formula lid by 26 neutralisation, for example by careful neutralisation 27 with a mineral acid such as hydrochloric, sulphuric or 28 nitric in aqueous solution, followed by extraction with 29 an appropriate organic solvent. Alternatively, the acids of general formula IIc may be obtained by 31 treating the compounds of general formula IId with an 32 ion exchange resin. If the acids of general formula IIc 33 are allowed to stand in solution they slowly WO 91/15482 PCT/GB91/00357 14 1 re-lactonise to the compounds of general formula I.

2 This process may be accelerated by heating a solution 3 of the acid under conditions that remove the water 4 formed, such as in a Dean-Stark apparatus, or by stirring the solution with a drying agent such as 6 anhydrous sodium sulphate, magnesium sulphate or 7 molecular sieves.

8 9 Lactones of general formula I may, if desired, be hydrolysed in the presence of an alcohol and a 11 catalytic amount of acid, preferably p-toluenesulphonic 12 acid, to produce compounds of general formula IIb.

13 14 A ketone of general formula IIa may be prepared by the methods outlined in Scheme in which R 1

R

4 and R 16 are as previously described, and P 1

P

2 and R 11 are 17 defined below.

18 19 Compounds of general formula IIa wherein b is a double bond may be prepared by removing the protecting groups 21 p 2 and P 3 from the compounds of general formula XIV.

22 This may be achieved in the preferred cases in which P 2 23 and P 3 are trialkylsilyl or alkyldiarylsilyl by the use 24 of conditions that generate fluoride anions, and preferably by using tetrabutylammonium fluoride in 26 tetrahydrofuran buffered with acetic acid or 27 hydrofluoric acid in aqueous acetonitrile.

28 29 Compounds of general formula IIa wherein b is a single bond may be obtained from compounds of general formula 31 IIa wherein b is a double bond by reduction of the 32 carbon-carbon double bond of the enone system, using 33 reagents and conditions that do not affect the other WO 91/15482 PCT/GB91/00357 1 functional groups present. Examples of such reagents 2 are sodium hydrogen telluride, triphenyltin hydride, or 3 tri-n-butyltin hydride with a palladium or platinum 4 catalyst.

6 Compounds of general formula IIa wherein b is a single 7 bond may also be prepared from enones of general 8 formula XIV by reduction of the double bond followed by 9 deprotection. For example it is possible to reduce the double bond by treatment of the enone with a 11 trialkylsilane, preferably triethylsilane, and a 12 catalyst such as tris(triphenylphosphine)rhodium 13 chloride [Wilkinson's catalyst] either neat, using an 14 excess of the silane, or in an inert hydrocarbon solvent such as benzene or toluene at a temperature 16 between ambient and reflux, preferably 50-70*C. The 17 crude silyl enol ether thus produced is treated with 18 hydrofluoric acid in aqueous acetonitrile to give the 19 compound of general formula IIa in which b is a single bond. However, the preferred method of transformation 21 of the compound XIV is to treat firstly with a reducing 22 agent such as sodium hydrogen telluride, or such 13 mixtures as tri-n-butyltin hydride with a palladium or 24 platinum catalyst and then deprotect in a second and separate reaction, in a manner similar to the 26 deprotection of the compound of general formula XIV.

27 28 Compounds of general formula lie may be prepared from 29 compounds of general formula IIa by hydrolysis with a base such as lithium hydroxide, sodium hydroxide or 31 potassium hydroxide in a mixture of water and an 32 organic solvent such as methanol, ethanol or 33 tetrahydrofuran at a temperature between 0OC and WO 91/15482 PCT/GB91/00357 16 1 preferably ambient temperature. The cation in compounds 2 of general formula IIe is usually determined by the 3 cation of the hydroxide employed; however, the cation 4 may then be exchanged for another cation by treatment with, for example, ion-exchange resins.

6 7 Compounds of general formula IIa may be used as 8 intermediates in the production of compounds of general 9 formulae IIb-e and of general formula I as detailed in Scheme I, or they may be used as HMG-CoA reductase 11 inhibitors in their own right.

12 13 An enone of general formula XIV may be prepared from an 14 aldehyde of general formula XII by reaction with a phosphonate of general formula XIII in which R 11 is a 16 lower alkyl group C 1 -8 or, preferably, C 1 -4 alkyl 17 grcup such as methyl or ethyl), and the group P 2 is any 18 group suitable for the protection of hydroxyl groups, 19 but preferably trialkylsilyl or alkyldiarylsilyl. The reaction between the aldehyde of general formula XII 21 and the phosphonate of general formula XIII is 22 preferably carried out in either of the following two 23 ways. In a first method the aldehyde of general formula 24 XII and phosphonate of general formula XIII are reacted together in the presence of a chelating metal halide 26 such as lithium chloride or magnesium bromide and a 27 mild organic base such as triethylamine or 28 1,8-diazabicyclo[4.5.0]undec-7-ene (DBU) in an inert 29 solvent such as acetonitrile or dimethylsulphoxide at ambient temperature. In a second method the phosphonate 31 of general formula XIII is first treated with a strong 32 organic base such as lithium diisopropylamide or 33 lithium or sodium bis(trimethylsilyl)amide in an inert WO 91/15482 PCT/GB91/00357 17 1 organic solvent such as diethyl ether or 2 tetrahydrofuran at a temperature between -78"C and 0*C, 3 the aldehyde of general formula XII added at the same 4 temperature, and the mixture allowed to warm to ambient temperature, all under an inert atmosphere.

6 7 An aldehyde of general formula XII may be prepared from 8 an alcohol of general formula X by conventional 9 oxidation reagents such as pyridinium chlorochromate or pyridinium dichromate, or by using a catalytic quantity 11 of tetra-n-propylammonium per-ruthenate and 12 N-methylmorpholine N-oxide, in an inert organic solvent 13 such as dichloromethane or tetrahydrofuran, but 14 preferably the oxidation is carried out using Swern's protocol.

16 17 An intermediate alcohol of general formula X may be 18 prepared for example in either of two ways from a diol 19 of general formula VII. In the first method the diol of general formula VII is di-acylated for example by 21 treatment with an excess of an acid anhydride 22 ((R 1

CO)

2 0)or acid halide (R 1 CO.Hal) in the presence of 23 a catalyst such as 4-(N,N-dimethylamino)pyridine, and a 24 base such as triethylamine or pyridine until both hydroxyl groups in the compound of general formula VII 26 have reacted. The diacylated compound of general 27 formula XI is then hydrolysed for example by treatment 28 with an alkali metal hydroxide such as lithium 29 hydroxide, potassium hydroxide or sodium hydroxide in a solvent such as water or an alcohol, or a mixture of 31 such solvents, at a temperature between O'C and ambient 32 for a time suitable to maximise the production of the 33 alcohol of general formula X.

WO 91/15482 PCT/GB91/00357 18 1 2 In the second and preferred of the two exemplary 3 methods the diol of general formula VII is treated 4 under conditions that will selectively protect the primary alcohol, either as an ester or an ether. Such 6 conditions are well known to one skilled in the art, 7 but the preferred conditions are to treat with one 8 equivalent of a trialkylsilylchloride in the presence 9 of imidazole and optionally, a mild organic base such as triethylamine or pyridine, and preferably using 11 dichloromethane or chloroform as a solvent. The product 12 of such a reaction will be a compound of general 13 formula VIII wherein P1 is a trialkylsilyl moiety or 14 other protective group. The compound of general formula VIII is then acylated, for example using the conditions 16 described above, that is treatment with the appropriate 17 acid halide (R 1 CO.Hal) or preferably the anhydride 18 ((R 1

CO)

2 0) using a mild organic base such as 19 triethylamine or pyridine and optionally using a catalyst such as 4-(N,N-dimethylamino)pyridine. The 21 resulting intermediate compound of general formula IX, 22 may then be deprotected to give the alcohol of general 23 formula X using such conditions as are appropriate for 24 the removal of the group P 1 without affecting the rest of the molecule. For the removal of the preferred 26 trialkylsilyl group, the preferred method is to treat 27 the protected alcohol of general formula IX with a very 28 mild acid or acid salt, for example pyridinium 29 p-toluenesulphonic acid, in a solvent such as methanol or ethanol. However, it will be appreciated by one 31 skilled in the art that other methods are available for 32 the removal of the preferred group, or that other 33 WO 91/15482 PCT/GB91/00357 19 1 protecting groups may be used in the transformation of 2 the diol of general formula VII to the alcohol of 3 general formula X.

4 Intermediate compounds of general formula XIV may also 6 be synthesised from the protected alcohols of general 7 formula XV using the sequence of reactions shown in 8 Scheme III, in which R 4

R

5

R

11 p 2 and P 3 are as 9 previously defined, and P 4 is define4 below.

11 An intermediate of general formula XIV may be prepared 12 from an enone of general formula XVIII by acylation for 13 example using conventional means. Thus a compound of 14 general formula XIV may be prepared by treating an alcohol of general formula XVIII with an acid halide 16 such as a chloride or bromide (RICO.Hal), or, 17 preferably, an anhydride ((R 1

CO

2 0) in the presence of a 18 mild organic base such as pyridine or triethylamine, 19 and preferably using a catalyst such as 4-(N,N-dimethylamino)pyridine, either neat or in an 21 inert solvent, preferably dichloromethane or chloroform 22 at a temperature between 0°C and reflux. Alternatively 23 the transformation may be carried out using the acid 24 (R 1 C0 2 H) and a coupling reagent such as a carbodiimide and a catalyst such as N,N-dimethylaminopyridine, in an 26 inert solvent and preferably at ambient temperature.

27 28 An enone of general formula XVIII may be prepared from 29 an aldehyde of general formula XVII and a phosphonate of general formula XIII as defined above, for example 31 by using a chelating metal halide such as lithium 32 chloride or magnesium bromide and a mild organic base 33 such as triethylamine or DBU in an inert organic WO 91/15482 PCT/GB91/00357 1 solvent preferably acetonitrile or dimethylsulphoxide, 2 at a temperature from 0° C to ambient and preferably 3 under an inert atmosphere.

4 To prepare an aldehyde of general formula XVII an 6 alcohol of general formula XV, in which the group P 4 is 7 any group suitable for the protection of alcohols, may 8 be oxidised to an aldehyde of general formula XVI for 9 example by conventional means such as pyridinium chlorochromate or pyridinium dichromate, or by using a 11 catalytic quantity of tetra-n-propylammonium 12 per-ruthenate (TPAP) in the presence of 13 N-methylmorpholine N-oxide in an inert solvent, 14 preferably dichloromethane, but most preferably by using Swern's protocol. The protecting group P 4 may 16 then be removed by any appropriate method known in the 17 art to give a hydroxy aldehyde of general formula XVII.

18 19 The intermediate alcohols VII and XV required for the syntheses outlined in reaction schemes II and III may 21 be prepared as shown in general reaction scheme IV, in 22 which R 5

P

3 and P 4 are as previously defined and R 1 0 23 is lower alkyl, preferably methyl or ethyl.

24 The intermediate alcohols of general formula VII may be 26 prepared by reduction of the ester group in the 27 compounds q. general formula XXII using conventional 28 reagents such as lithium aluminium hydride, 29 di-isobutylaluminium hydride or lithium triethylborohydride in an inert organic solvent such as 31 diethyl ether or tetrahydrofuran, at ambient 32 33 WO 91/15482 PCT/GB91/00357 21 1 temperature to reflux, under an inert atmosphere. The 2 alcohols of general formula VII may be then be used as 3 outlined in scheme II.

4 An intermediate of general formula XV may be obtained 6 from an ester of general formula XXII by first 7 protection and then reduction. The protective group P4 8 may be any ether group known in the art for the 9 protection of alcohols, for example trialkylsilyl, alkoxymethyl, benzyl, or substituted benzyl ethers, and 11 may be introduced using the conventional methods for 12 such groups. The reduction of the ester group may then 13 be carried out using the conventional methods described 14 above, to yield the alcohol of general formula XV which may then be used as outlined in scheme III.

16 17 The intermediate of general formula XXII may be 18 prepared in two steps from the ester of general formula 19 XXVII. In the first step the acetate group is removed, for example by ester exchange using either acidic or 21 basic conditions, but preferably by treatment with a 22 solution of sodium ethoxide in ethanol at ambient 23 temperature. In the second step the hydroxyl group 24 released in the first step is protected with the group

P

3 where P 3 is as previously defined. This 26 transformation may be carried out in any suitable 27 manner known in the art, but in the preferred case in .28 which P 3 is an alkyldiarylsilyl group, the reaction is 29 preferably performed by treating the alcohol with an alkyldiarylsilylchloride in the presence of a mild base 31 such as imidazole, in a solvent such as dimethyl- 32 formamide, chloroform or dichloromethane.

33 WO 91/15482 PCT/GB91/00357 22 1 The intermediate acetate of general formula XXVII may 2 Le obtained from the methyl ketone of general formula 3 XXV by means of, for example, the Baeyer-Villiger 4 reaction. The application of this reaction to the synthesis of novel compounds constitutes an important 6 aspect of this invention. For example, the ketone of 7 general formula XXV may be treated with a per-acid such 8 as peracetic acid, trifluoroperacetic acid or 9 trimethylsilyl peroxide in an inert solvent such as dichloromethane or chloroform. Optionally the double 11 bond (if present) in the compound of general formula 12 XXV may be protected prior to the oxidation reaction, 13 for example by reacting with bromine to give the 14 dibromo-derivative; after oxidation of the methyl ketone group in a manner similar to that described 16 above the double bond may be restored by treatment of 17 the product of oxidation with zinc in acetic acid to 18 yield the acetate of general formula XXVII.

19 The methyl ketone of general formula XXV may be 21 obtained from the lactone of general formula XXIII for 22 example by either of the two following methods. In a 23 first method the keto-sulphone of general formula XXIV 24 may be produced from the lactone of general formula XXIII by reaction with an anion or dianion of methyl 26 sulphone in an inert organic solvent, preferably 27 tetrahydrofuran, at to ambient temperature under 28 an inert atmosphere. The methyl ketone may then be 29 obtained by treating the keto-sulphone with a reagent that can selectively remove the sulphone group, such as 31 Raney nickel, sodium amalgam or aluminium amalgam in a 32 solvent such as an alcohol, or by treatment with 33 trib-tyltin hydride.

WO 91/15482 PCT/GB91/00357 23 1 2 In a second and preferred method the lactone of general 3 formula XXIII is reacted with the anion of tert-butyl 4 acetate (generated by conventional means) in a solvent such as tetrahydrofuran or diethyl ether under an inert 6 atmosphere at a temperature from -70*C to ambient. The 7 ketone c. general formula XXVI thus obtained may then 8 be treated with a strong acid such as trifluoroacetic 9 acid in a solvent such as dichloromethane or tetrahydrofuran optionally containing a little water, 11 at room temperature, to give the methyl ketone of 12 general formula XXV.

13 14 The ketone of general formula XXV may be used as obtained from either of the two methods outlined above, 16 or if desired the stereochemistry of the carbon atom to 17 which the methyl ketone group is attached may be 18 altered by treatment with a mild base such as a sodium 19 alkoxide in an alcoholic solvent. This treatment may give a product of general formula XXV in which the 21 methyl ketone group is mainly in the equatorial 22 position and only small amount in the axial position.

23 24 Intermediates of general formula XXIII in which R 5 is methyl, R 10 is ethyl, and a is a double bond, are known 26 in the literature Chem. Soc., Chem. Commun., 1987, 27 1986). Those intermediates in which R 5 and R 1 0 are 28 other groups within the appropriate definitions may be 29 prepared using routes analogous to the known route, but using the appropriately different starting materials.

31 Such a change is within the scope of one skilled in the 32 art. Methods for the reduction to give a as a single 33 bond in compounds with structures similar to the WO 91/15482 PCT/GB91/00357 24 1 compounds of general formulae I, II, VII XII, and XIV 2 XXVII are known in the art (for examples, see 3 Tetrahedron 1986, 42, 4909-4951 or US-A-4826999). Some 4 of these methods may use reagents that deleteriously affect the compounds of general formulae I, II, VII 6 XII, and XIV XXVII; however, other methods may be 7 suitable for the required transformations in some or 8 all of the compounds of general formulae I, II, VII 9 XII, and XIV XXVII. Thus it is within the capabilities of one skilled in the art to select 11 appropriate methodology for the interconversion of 12 compounds wherein a may be a single or double bond in 13 order to obtain the compounds of general formula I or 14 II with the required single or double bond at a.

16 A phosphonate of general fcrmula XIII in which R 4 and 17 R 1 1 are methyl and P2 is a t-butyldimethylsilyl group 18 is known in the art Orq. Chem., 1988, 53, 19 2374-2378).

21 In general, reagents are used in sufficient quantities 22 to completely convert starting materials to products 23 but to be themselves substantially consumed during the 24 course of the reaction. However the amounts may often be varied as is evident to one of ordinary skill in the 26 art. For example, in a reaction of two compounds one of 27 which is not readily available and one of which is, an 28 excess of the readily available compound may be used to 29 drive the reaction further towards completion (unless the use of an excess would increase the synthesis of an :1 undesired compound).

32 33 WO 91/15402f PCI7/C 01191/00357 1 Likewise, most of the temperature ranges given in the 2 preceding descriptions are merely exemplary, and it is 3 within the ability of one of ordinary skill in the art 4 to vary those that are not critical.

6 The reaction times set forth in the preceding 7 description are also merely exemplary and may be 8 varied. As is well-known, the reaction time is often 9 inversely related to the reaction temperature.

Generally, each reaction is monitored by, for example, 11 thin layer chromatography and is terminated when at 12 least one starting material is no longer present, or 13 when it appears that no more of the desired product is 14 being formed.

16 Conventional work-up procedures have generally been 17 omitted from the preceding descriptions.

18 19 As utilised in the preceding descriptions, the term "nolvent" embraces mixcures of solvents and implies 21 that the reaction medium is a liquid at the desired 22 reaction temperature. It should, therefore, be 23 understood that not all of the solvents listed for a 24 particular reaction may be utilised for the entire cited temperature range. It should also be understood 26 that the solvent must be at least substantially inert 27 to the reactants employed, intermediates generated and 28 end products under the reaction conditions utilised.

29 The term "inert atmosphere", as utilised in the 31 preceding descriptions, means an atmosphere that does 32 not react with any of the reactants, intermediates or 33 end products or otherwise interfere with the reaction.

WO 91/15482 PCl'/GB91/00357 26 1 While a carbon dioxide atmosphere is suitable for 2 certain reactions, the inert atmosphere is usually 3 nitrogen, helium, neon, or argon, or a mixture thereof, 4 and most often dry argon to maintain anhydrous ccnditions. Most reactions, including those where the 6 use of an inert atmosphere is not specified, are 7 carried out under an inert atmosphere, usually dry 8 argon, for convenience.

9 The product of each reaction may, if desired, be 11 purified by conventional techniques such as 12 recrystalisation (if a solid), column chromatgraphy, 13 preparative thin layer chromatography, gas 14 chromatography (if sufficiently volatile), fractional distillation under high vacuum (if sufficiently 16 volatile) or high pressure (performance) liquid 17 chromatography (HPLC). Often, however, the crude 18 product of one reaction may be employed in the 19 following reaction without purification or even isolation.

21 22 Some reactions, particularly those utilizing strong 23 bases or reducing agents, require anhydrous solvents.

24 Where this is the case solvents may be dried before use using conventional techniques and an inert atmosphere 26 used.

27 28 Some of the reactions described above may yield 29 mixtures of two or more products, only one of which leads to the desired compound of general formula I or 31 II. Any mixture so obtained may be separated by 32 conventional techniques such as those set forth in the 33 preceding paragraphs.

WO 91/15482 PCT/GB91/00357 27 1 2 Certain of the intermediate componds described above 3 are believed to be novel, in particular compounds of 4 general formulae VII-XII, XIV-XXII, and XXV-XXVII. All other intermediate compounds in which R 5 is not methyl 6 are also believed to be novel. Novel intermediates and 7 processes for preparing them form further aspects of 8 this invention.

9 Compounds of this invention are useful as 11 antihypercholesterolaemic agents for the treatment of 12 arteriosclerosis, hyperlipidaemia, familial 13 hypercholesterolaemia and the like diseases in humans.

14 According to a third aspect of the invention, there is 16 therefore provided a compound of general formula I or 17 II for use in medicine, particularly as 18 antihypercholesterolaemic agents.

19 According to a fourth aspect of the invention, there is 21 provided the use of a compound of general formula I or 22 II in the preparation of an antihypercholesterolaemic 23 agent. Compounds of the invention can therefore be used 24 in a method for the treatment or prophylaxis of hypercholesterolaemia in general and arteriosclerosis, 26 familial hypercholesterolaemia or hyperlipidaemia in 27 particular comprising administering to a patient an 28 effective dose of a compound of general formula I or II 29 or a mixture thereof.

31 According to a fifth arpect of the invention, there is 32 provided a pharmaceutical composition comprising a 33 compound of general formulae I or II, or a mixture O 91/15482 PCT/G1391/00357 28 1 thereof, and a pharmaceutically acceptable carrier.

2 Such a composition may simply be prepared by the 3 admixture of the ingredients.

4 Compounds of general formulae I and II may be 6 administered orally or rectally or parenterally in the 7 form of a capsule, a tablet, an injectable preparation 8 or the like. It is usually desirable to use the oral 9 route. Doses may be varied, depending on the age, severity, body weight and other conditions of human 11 patients but daily dosage for adults is within range 12 of from about 2 mg to 2000 mg (preferably 5 to 100 mg) 13 which may be given in one to four divided doses. Higher 14 doses may be favourably employed as required.

16 The compounds of this invention may also be 17 co-administered with pharmaceutically acceptable 18 nontoxic cationic polymers capable of binding bile 19 acids in a non-reabsorbable form in the gastrointestinal tract. Examples of such polymers 21 include cholestyramine, colestipol and poly[methyl-(3- 22 trimethylamino-propyl)iminotrimethylene dihalide]. The 23 relative amounts of the compounds of this invention and 24 these polymers is between 1:100 and 1:15000.

26 The intrinsic HMG-CoA reductase inhibition activity of 27 the claimed compounds may be measured in in vitro 28 protocols described in detail in the Examples below.

29' Included within the scope of this invention is the 31 method of treating arteriosclerosis, familial 32 hypercholesterolaemia or hyperlipidaemia which 33 comprises administering to a subject in need of such WO 91/15482 PCI/G B91/00357 29 1 treatment a nontoxic therapeutically effective amount 2 of the compounds of general formulae I or II or 3 pharmaceutical compositions thereof.

4 The following examples show representative compounds 6 encompassed by this invention and their syntheses.

7 However, it should be understood that they are for the 8 purposes of illustration only.

9 Organic solutions were dried over sodium sulphate or 11 magnesium sulphate, and evaporated under reduced 12 pressure. NMR spectra were recorded at ambient 13 temperature in deuteriochloroform at 250 MHz for proton 14 and 62.5 MHz for carbon unless noted otherwise. All chemical shifts are given in parts per million relative 16 to tetramethylsilane. Infra red spectra were recorded 17 at ambient temperature in solution in chloroform, or in 18 the solid state in a potassium bromide disc as noted, 19 and are are expressed in reciprocal centimeters.

21 Chromatography was carried out using Woelm 32-60 jim 22 silica.

23 24 26 27 28 29 31 32 33 WO 91/15482 WO 9115482PCT/GB191/00357 1 Example 1 2 3 Methyl 7-(1-F(lS, 2S, 4aR. 6R, _8S, -8a)-6-hydroxcy-8- 4 r,(2(S)-methyl-1-oxobutvl)oxyl-1, 2. -4gl. 5. 6, 7, 8, 8a-octahydro-2-methvlnaphthalenll]1-3(R) 6 Wrdroxvheptanoate 7

HO

a

CO

2 Me 9 OH 12 2H Me 13 14

HO

16 17 Step 1 18 19 t-Butyl 2S, 4aR, 6S, 8S, 8aS)-6-(1-ethoxy- 2n carbonyl-1, 2, 4a, 5, 6, 7, 8, Ba-octahydro-8-hydroxy- 21 2-methylnaphthalenyl)])-3-oxopropiQnate (XXVI) 22 23 A solution of freshly distilled tert-bu..yl acetate (4.4 24 g, 38 iimmole) in dry tetrahydrofuran (5 mL) was added dropwise to a well stirred solution of lithium 26 hexamethyldisilazide in THF (1.0 M; 38 mmole) at -78*C 27 under an argon atmosphere. After 3.5 hours, a solution 28 of (+)-Ethyl (1S, 2S, 4aR, 6S, 8S, 8aS)-1, 2, 4a, 5, 6, 29 7, 8, 8a-octahydro-2-methyl-6,8-naphthalenecar-bolactone-1-carboxylate (XXIII; R 5 represents methyl, Rio 31 represents ethyl) (2.0 g, 7.58 mmole) in dry TH-F (5 mL) 32 was added dropwise, the solution stirred for 90 minutes 33 at -700C then quenched with a saturated solution of WO 91/15482 WO 9115482PCIT/G191 /00357 31 1 ammonium chloride (15 mL) The mixture was warmed to 2 room temperature, then separated and the aqueous phase 3 extracted with dichioromethane (2 x 40 mL). The 4 combined organic phases were washed with brine (20 mL) then dried and evaporated to leave a gum (3.34 g) which 6 was purified by coliumn chromatography eluting with 4:1 7 hexane:ethyl acetate to 1:1 ethyl acetate:hexane, to 8 give the title compound (1.6 g, 56%).

9 delta H 5.50 (1H, ddd, J 9.8, 4.5 and 2.5 Hz, 3-H) 11 5.31 (1H, d, a 9.8 Hz, 4.16 (111, m, 4.07 12 (2H1, m, OCH 2 3.69 (1H1, m, OH), 3.44 (2H, d, -J 1.8 Hz, 13 COCH 2 CO), 2.95 (1H1, t, Lj. 6.3 Hz, 2.76 (1H1, dd, J 14 11.6 and 6.0 Hz, 2.52 (1H, m, 2.18 (1H1, br d, J 16.3 Hz, 7-Hax) 2.13 2.0 (2H, m, 4a-H and 16 5-Hax), 1.78 (1H, ddd, J~ 15.2, 6.4 and 3.5 Hz, 7-Heq), 17 1.45 1.28 (11H, m, :L-Bu, 8a-H, and 5-Hax), 1.18 (3H, 18 t, J 7 Hz, CH 2 Cfl 3 and 0.82 (3H, d, J 7 Hz, CHCUj 3 19 delta C 210.0, 173.3, 166.3, 131.4, 129.6, 81.9, 64.7, 21 59.8, 48.1, 46.0, 44.4, 40.1, 32.5, 32.2, 32.0, 29.4, 22 27.8, 17.4 and 14.1.

23 24 Step 2 26 27 Ethyl (iS, 2S, 4aR, 6R, 8S, 8aS) 2, 4a, 5, 6, 7, 8, 28 8a) -Octahydro-8-hydroxy-2-methyl-6- (1-oxoethyl) naphtha- 29 lenyl-1-carboxylate (XXV) 31 T*:ifluoroacetic acid (3.75 nmL, 48.7 mmole) was added to 32 u± stirred solution of the keto-ester (XXVI) prepared in 33 step 1 (1.5 g, 3.9 mmole) in dichioromethane (4 mL) WO 91/15482 PCT/GB91/00357 32 1 followed by water (0.3 mL). The solution was stirred 2 for 75 minutes, then brine (10 mL) and dichloromethane 3 (10 mL) added, the aqueous phase separated and 4 extracted with dichloromethane (3 x 10 mL). The combined organics were washed with a saturated sodium 6 bicarbonate solution (20 mL) and brine (10 mL), dried 7 and evaporated to give a solid. The solid (1.1 g) was 8 dissolved in sodium ethoxide solution (prepared by 9 dissolving sodium (70 mg, 3.04 mmole) in absolute ethanol (20 mL)) and after 1 hour the solvent was 11 evaporated to give a gum, which was partitioned between 12 brine (20 mL) and dichloromethane (50 mL). The aqueous 13 phase was separated, extracted with dichloromethane 14 mL) and the combined organic phases washed with brine (20 mL), dried and evaporated to leave a gum, which was 16 purified by column chromatography eluting with 1:2 17 ethyl acetate:hexane to give the title ketone (XXV) 18 (0.945 g, 87%).

19 nu max. (KBr disc) 3700 3100, 1730, 1705.

21 22 delta H 5.57 (1H, ddd, J 9.8, 4.4 and 2.7 Hz, 3-H), 23 5.42 (1H, d, J 9.8 Hz, 4.43 (1H, br s, 4.23 24 4.07 (2H, m, OCH2), 2.9 (1H, tt, J 12.6 and 3.7 Hz, 2.84 (1H, dd, 3 11.6 and 5.9, 2.64 (1H, m, 26 2.40 (1H, m, 4a-H), 2.16 (3H, s, CH 3 CO), 2.15 27 1.9 (2H, m, 5-Heq and 7-Heq), 1.68 1.32 (3H, 1.27 28 (3H, t, J 7 Hz, CH 2

CH

3 1.25 (1H, m) and 0.93 (3H, d, 29 J 7 Hz, CHCH3),.

31 delta C 210.0, 173.5, 131.0, 130.0, 65.9, 60.0, 45.4, 32 44.9, 39.4, 35.4, 34.3, 33.2, 32.4, 28.0, 17.3 and 33 14.2.

WO 91/15482 PCT/GB91/00357 33 1 2 Step 3 3 4 Ethyl (1R, 2R, 3S, 4S, 4aS, 6R, 8S, 8aR) 3,4-dibromodecahydro-8-hydroxy-2-methyl-6-(l-oxoethyl)- 6 naphthalenyl-l-carboxylate 7 8 A solution of bromine (0.54 g, 3.4 mmole' in carbon 9 tetrachloride (25 mL) was added dropwise to a cold stirred, solution of the compound of step 2 11 (XXV) (0.94 g, 3.37 mmole) in a mixture of the same 12 jolvent (25 mL) and absolute ethanol (1.5 mL), under an 13 argon atmosphere. The pale orange solution was stirred 14 for 10 minutes then washed with an aqueous solution of NaHSO 3 followed by a saturated solution of sodium 16 hydrogen carbonate (30 mL). The organic solution was 17 dried and evaporated to leave the dibromide (1.61 g) as 18 a solid, which was used directly in the next step.

19 delta H 4.87 (1H, m, CHBr), 4.53 (1H, m, CHBr), 4.35 21 (1H, m, 4.22 4.09 (2H, m, OCH 2 3.40 (1H, dd, 22 J 11.6 and 5.1 Hz, 2.99 (1H, tt, J 12.3 and 3.6 23 Hz, 2.77 (1H, m, 2.49 (1H, ct, J 11.3 and 24 3.6 Hz, 4a-H), 2.16 (3H, s, CH 3 CO), 2.15 1.54 1.32 (3H, d, J 7 Hz, CHCH 3 and 1.26 (3H, t, J 7.2 26 Hz, CH 2

CH

3 27 28 delta C 211.5, 173.1, 65.5, 60.3, 57.5, 57.4, 43.9, 29 42.0, 39.2, 35.2, 35.0, 33.7, 32.1, 27.9, 19.5 and 14.1.

31 32 33 WO 91/15482 PCT/GB91/00357 34 1 Step 4 2 3 Ethyl (1R, 2R, 3S, 4S, 4aS, 6R, 8S, 8aR) 6-acetoxy-3,4- 4 dibromodecahydro-8-hydroxy-2-methylnaphthalenyl-lcarboxylate 6 7 Trifluoroacetic anhydride (7.75 mL, 55.5 mmole) was 8 added to a well-stirred mixture of hydrogen peroxide 9 solution (60% w/v, 1.2 mL, 20.8 mmole) in dichloromethane (5 mL) at O'C. After 15 minutes, the 11 clear solution was added rapidly to a stirred ice-cold 12 solution of the crude dibromide from the previous step 13 (1.52 g) in dichloromethane (6 mL), then the cooling 14 bath was removed. After 30 minutes, more dichloromethane (25 mL) was added, the solution 16 re-cooled to 0°C and saturated sodium bicarbonate 17 solution added slowly ui.til the mixture reached pH 8.

18 The aqueous phase was separated, extracted with 19 dichloromethane (2 x 25 mL) and the combined organic extracts dried and evaporated to leave the acetate, 21 (1.37 g) as a white foam.

22 23 delta H (key peaks) 5.17 (1H, tt, J 11.5 and 4.4 Hz, 24 6-H) and 2.02 (3H, s, CH 3

CO

2 26 27 Step 28 29 Ethyl (lS, 2S, 4aR, 6R, 8S, 8aS) 6-acetoxy-l, 2, 4a, 6, 7, 8, 8a-octahydro-8-hydroxy-2-methylnaphthalenyl-l- 31 carboxylate (XXVII) 32 33 WO 91/15482 PCT/GB91/ 00357 1 Freshly prepared active zinc (1.91 g, 29.2 mmole) was 2 added in several portions to a stirred solution of the 3 crude acetate from the previous step (1.33 g) and 4 acetic acid (0.5 mL) in dry ether (15 mL). After the refluxing had subsided, the mixture was stirred for 6 minutes, then filtered and the solid washed with ether.

7 The filtrate was washed with saturated sodium 8 bicarbonate solution, dried and evaporated. The residue 9 was purified by column chromatography eluting with 1:4 ethyl acetate:hexane, to give the title compound (645 11 mg, 65% from XXV).

12 13 delta H (key peaks) 5.55 (1H, ddd, J 9.8, 4.1 and 2.7 14 Hz, 5.39 (1H, d, J 9.8 Hz, 4-H) and 5.12 (1H, tt, J 11.5, 4.5, 6-H).

16 17 delta C 173.5, 170.5, 131.0, 129.7, 69.3, 67.0, 60.0, 18 44.6, 39.2, 37.8, 32.4, 32.2, 21.2, 17.4 and 14.2.

19 21 Step 6 22 23 Ethyl (1S, 2S, 4aR, 6R, 8S, 8aS) 1, 2, 4a, 5, 6, 7, 8, 24 8a-octahydro-6,8-dihydroxy-2-methylnaphthalenyl-lcarboxylate 26 27 The acetate (XXVII) prepared in step 5 (615 mg, 2.08 28 mmole) was dissolved in a solution of sodium ethoxide 29 (8 mL), prepared using sodium metal (50.2 mg, 2.18 mmole). After 30 minutes, the solvent was evaporated 31 and the residue partitioned between brine (25 mL) and 32 dichloromethane (25 mL). The brine was extracted with 33 WO 91/15482 PCT/GB91/00357 36 1 more dichloromethane (2 x 25 mL) and the combined 2 organic extracts dried and evaporated to leave the 3 alcohol as a solid (0.476 g, 4 delta H (key peaks) 4.05 tt, 11.3 and 4.5 Hz, 6 6-H).

7 8 delta C 173.6, 130.9, 130.1, 67.4, 66.3, 60.0, 44.8, 9 42.9, 41.8, 39.2, 32.5, 17.4, 14.2.

11 12 Step 7 13 14 Ethyl (IS, 2S, 4aR, 6R, 8S, 8aS) 6-t-butyldiphenylsilyloxy-1, 2, 4a, 5, 6, 7, 8, 8a-octahydro-2methyl- 16 naphthalenyl-l-carboxylate (XXII) 17 18 Tert-butyldiphenylsilylchloride (0.44 mL, 1.71 mmole) I3 was added dropwise to a stirred solution of the diol prepared in the previous step (416 mg, 1.64 mmole) and 21 imidazole (223 mg, 3.27 mmole) in dry DMF (3 mL), under 22 an argon atmosphere. After 16 hours, brine (10 mL) was 23 added, and the mixture extracted with ether (2 x 24 mL), which was then washed sequentially with 2M hydrochloric acid (10 mL), water (2 x 10 mL) and 26 saturated sodium bicarbonate solution (10 mL). The 27 organic solution was dried and evaporated to leave the 28 crude title compound (XXII) (740 mg).

29 delta H (key peaks) 7.74 7.32 (10H, m, Ph) and 1.06 31 (9H, s, t-butyl).

32 33 WO 91/15482 PCT/GB91/00357 37 1 Step 8 2 3 (IS, 2S, 4aR, 6R, 8S, 8aS) 6-t-Butyldiphenylsilyloxy-1, 4 2, 4a, 5, 6, 7, 8, 8a-octahydro-l-hydroxymethyl-2methylnaphthalene (VII) 6 7 A solution of the ester (XXII) prepared in the previous 8 step (0.74 g) in dry ether (6 mL) was added dropwise to 9 a solution of lithium aluminium hydride (0.19 g, 4.92 mmole) in dry ether (5 mL), cooled in a cold water 11 bath. After 1 hour water (0.2 mL) was added slowly to 12 the stirred, cooled mixture, followed by aqueous sodium 13 hydroxide (15 0.2 mL) then more water (0.6 mL). The 14 mixture was filtered and the filtrate evaporated to leave a gum (0.74 which was purified by column 16 chromatography eluting with 1:2 ethyl acetate:hexane to 17 give the title diol (VII) (0.67 g, 91%).

18 19 nu max. (KBr) 3660 3100, 3100 3000.

21 delta H (key peaks) 3.67 3.52 (2H, m, CH2OH).

22 23 delta C 135.7, 134.7, 134.6, 131.9, 131.0, 129.4, 24 127.4, 68.0, 67.8, 64.9, 42.4, 42.2, 42.0, 40.9, 34.5, 33.7, 27.0, 19.1 and 15.5.

26 27 28 Step 9 29 (IS, 2S, 4aR, 6R, 8S, 8aS)-1-(t-Butyldimethylsilyloxy- 31 methyl)-6-(1-butyldiphenylsilyloxy)-1, 2, 4a, 5, 6, 7, 32 8, 8a-octahydro-8-hydroxy-2-methylnaphthalene (VIII) 33 WO 91/15482 W CY/G1391/00357 38 1 t-Butyldimethylsilylchloride (0.228g, 1.51 mmol) was 2 added in portions to a stirred solution of the diol 3 (VII) prepared in step 8 (0.622g, 1.38 mmol) in 4 dichloromethane (10 mL) under an atmosphere of argon.

The mixture was stirred for 18 hours then more 6 dichloromethane (20 mL) added and the organic phase 7 washed successively with 1M H 3

PO

4 (10 mL), sodium 8 hydrogen carbonate solution (10 mL) and brine (10 mL).

9 The dichloromethane was dried and evaporated to leave a gum, which was purified by column chromatography 11 eluting with 19:1 hexane:ethyl acetate to give the 12 title compound as a gum (0.668g, 86%).

13 14 delta H (key peaks) 0.89 (9H, s, SiC(CH 3 3 0.076 (3H, s, SiCH 3 and 0.073 (3H, s, SiCH 3 16 17 18 Step 19 2S, 4aR, 6R, 8S, 8aS)-1-(L-Butyldimethylsilyl- 21 oxymethyl)-6-(-butyldiphenylsilyloxy)-1, 2, 4a, 5, 6, 22 7, 8, 8a-octahydro-2-methylnaphthalenyl] 2(S)-methyl- 23 butyrate (IX) 24 A solution of the alcohol (VIII) prepared in step 9 26 (0.668g, 1.18 mmol), 2-S methylbutyric anhydride (1.32 27 g, 7.1 mmol), 4-(dimethylamino)pyridine (22 mg) and dry 28 pyridine (2.00 mL, 24.9 mmol) in dichloromethane 29 mL) was stirred for 26 hours under argon. Methanol mL) was added and after stirring for 2 hours, diethyl 31 ether (100 mL) added and the solution washed with 1M 32 H 3

PO

4 (30 mL), water (2 x 25 mL) and saturated sodium 33 hydrogen carbonate solution (25 mL). The organic WO 91/15482 Per/cm 111/00357 39 1 solution was dried and evaporated, to leave the ester 2 (IX) as a gum (0.734g) which was used in the next step 3 without further purification.

4 delta H (key peaks) 5.03 (1H, m, 2.15 (1H, 6 obscured sextet, J 6.9 Hz, CO.CH), 1.6 1.2 (2H, 7 obscured multiplet, CH2CH 3 0.97 (3H, d, J 6.8 Hz, 8 CO.CHCH3) and 0.80 (3H, t, J. 7.4 Hz, CH 2

CH

3 9 11 Step 11 12 13 2S, 4aR, 6R, 8S, 8aS)-6-(t-Butyldiphenylsilyl- 14 oxy)-1, 2, 4a, 5, 6, 7, 8, 8a-octahydro-l-hydroxymethyl-2-methylnaphthalenyl] 2(S)-methylbutyrate (X) 16 17 The prctected alcohol (IX) prepared in step 10, was 18 dissolved in a solution of tosic acid monohydrate (200 19 mg, 1.07 mmol) in methanol (250 mL). After 2 hours, saturated sodium hydrogen carbonate solution (100 mL) 21 was added, the mixture stirred for 5 minutes then 22 concentrated to about 30 mL volume. The residue was 23 extracted with dichloromethane (3 x 100 mL); the 24 dichloromethane dried and evaporated to leave a gum (0.728g) which was purified by column chromatography 26 eluting with 9:1 hexane:ethyl acetate to 4:1 27 hexane:ethyl acetate to give the title compound (418 28 mg) plus unreacted started material (IX) (137 mg).

29 The unreacted material (IX) (137 mg) was recycled using 31 the same ratio of reagents and purified by column 32 chromatography as above to give a further 68 mg of the 33 title compound, a combined total yield of 486 mg WO 9111541822 W 1C'/GB9I/00357 1 2 delta H (key peaks) absence of signals at 0.87, 0.053 3 and 0.023 for t-butyldimethyylsily group.

4 delta C 175.7, 135.6, 134.4, 134.1, 132.6, 129.7, 6 129.5, 127.4, 69.8, 68.2, 61.8, 41.9, 41.3, 40.0, 39.3, 7 39.2, 34.8, 31.5, 26.9, 26.2, 19.0, 16.3, 15.4 and 8 11.4.

9 11 Step 12 12 13 2S, 4aR, 6R, 8S, 8aS)-6-(t-Butyldiphenylsilyl- 14 oxy)-1-formyl-1, 2, 4a, 5, 6, 7, 8, 8a-octahydro- 2-methylnaphthalenyl)] 2(S)-methylbutyrate (XII) 16 17 A solution of dry DMSO (178 mg, 2.28 mmol) in dry 18 dichloromethane (0.7 mL) was added to a stirred 19 solution of oxalylchloride (145 mg, 1.14 mmol) in dry dichloromethane (1.8 mL) under argon, at -70C. After 21 minutes, a solution of the alcohol prepared in step 22 11 (486 mg, 0.91 mmol) in dry dichloromethane (1.8 mL) 23 was added rapidly. After a further 10 minutes a second 24 batch of "activated DMSO" (prepared as above) was added to the stirred mixture, then after an additional 26 minutes dry triethylamine (1.14 mL, 8.2 mmol) was added 27 rapidly. After 5 minutes more at -70*C, the solution 28 was allowed to warm to room temperature, diethyl ether 29 (50 mL) was added and the organic phase washed successively with 1M H 3

PO

4 (20 mL), water (2 x 20 mL), 31 saturated sodium hydrogen carbonate solution (20 mL) 32 and dried. Evaporation gave the crude aldehyde (XII) 33 WO 91/15482 PCT/GB91/00357 41 1 (513 mg) which was purified by column chromatography 2 eluting with 1:15 ethyl acetate:hexane) to give the 3 title compound (398 mg, 82%).

4 delta H (key peaks) 9.69 (1H, d, J 2.5 Hz, CHO), 2.52 6 (1H, ddd, J 11.3, 5.7 and 2.5 Hz, 1-H).

7 8 delta C (key peak) 203.2.

9 11 Step 13 12 13 Methyl 2R, 4aR, 6R, 8S, 8aS)-6-(-butyldi- 14 phenylsilyloxy)-8-[(2(S)-methyl-l-oxobutyl)oxy-1l, 2, 4a, 5, 6, 7, 8, 8a octahydro-2-methylnaphthylenyl]) 16 -3(R)-(t-butyldimethylsilyloxy)-5-oxohepten-6-oate 17 (XIV) 18 19 Lithium bis(trimethylsilyl)amide in THF (1.0 M; 1.42 mmol) was added slowly to a stirred solution of methyl 21 3(R)-(t-butyldimethylsilyloxy)-6-(dimethoxyphosphonyl)- 22 -5-oxohexanoate (XIII; R 4 and R 1 1 both represent 23 methyl, P 2 represents the t-butyldimethylsilyl group) 24 (0.68 g, 1.78 mmol) in THF (5 mL) at -70C, under an atmosphere of argon. After 1 hour, a solution of the 26 aldehyde (XII) prepared in the previous step (378 mg, 27 0.71 mmol) in THF (5 mL) was added dropwise, the 28 solution warmed to room temperature, and stirred for 66 29 hours. Saturated ammonium chloride solution (5 mL) was added and the mixture extracted with dichloromethane (3 31 x 25 mL). The combined dichloromethane extracts were 32 33 WO 91/15482 PCT/GB91/00)357 42 1 dried, evaporated and the residue purified by column 2 chromatography eluting with 19:1 hexane: ethyl acetate 3 to give the title compound (380 mg, 68%).

4 delta H (key peaks) 6-74 (1H, dd, J 15.75 and 10.2 Hz, 6 5.92 (1H, d, 15.75 Hz, 4.89 (1H, m, 7 4.60 (1H, pentet, J 6.3 Hz, 3.65 (3H, s, 8 CO 2 Me), 2.74 and 2.49 2 x 2H, H-2' and and 9 1.02 (9H, s, SiC(CH 3 3 11 delta C 197.1, 174.6, 171.4, 147.9, 135.6, 134.3, 12 133.9, 131.9, 131.8, 129.9, 129.5, 127.4, 70.2, 68.1, 13 65.9, 51.3, 47.5, 42.7, 42.4, 41.7, 41.1, 40.4, 39,7, 14 35.7, 34.0, 26.8, 26.1, 25.6, 19.0, 17.8, 16.5, 16.3, 11.7, -4.8 and -5.1.

16 17 18 Step 14 19 Methyl 2S, 4aR, 6R, 8S, 8aS)-6-hydroxy- 21 8-[(2(S)-methyl-l-oxobutyl)oxy]-l, 2, 4a, 5, 6, 7, 8, 22 8a-octahydro-2-methylnaphthalenyl]}-3(R)-hydroxy-5-oxo- 23 heptanoate (IIa) 24 A mixture of tellurium (0.137g, 1.075 mmol) and sodium 26 borohydride (93 mg, 2.47 mmol) in deoxygenated ethanol 27 (5 mL) was heated to reflux, under argon, for 1 hour.

28 The purple solution was cooled and finely ground 29 ammonium chloride (0.58 g, 10.8 mmol) added, fol:rwed by a solution of the enone (XIV) prepared in step 13 31 (340 mg, 0.43 mmol) in deoxygenated ethanol (5 mL).

32 After stirring for 2 hours, a second batch of sodium 33 hydrogen telluride was prepared (on one fifth the scale WO 91/15482 PCI'/G 191/00357 43 1 above) and added to the reaction, together with more 2 ammonium chloride (116 mg, 2.16 mmol). The mixture was 3 stirred for a further 2 hours then saturated ammonium 4 chloride solution (2 mL) added. The mixture was concentrated to a small volume, extracted with 6 dichloromethane (3 x 20 mL), and the dichloromethane 7 dried and evaporated to leave a gum (355 mg).

8 9 A solution of the gum (315 mg) in 1:19 40% aq HF:acetonitrile (20 mL) was stirred for 6 hours at room 11 temperature. Saturated sodium hydrogen carbonate 12 solution (50 mL) was added carefully and the mixture 13 concentrated under reduced pressure, then extracted 14 with ethyl acetate (3 x 100 mL). The combined organic extracts were dried and evaporated to leave a crude gum 16 (220 mg), which was purified by column chromatography 17 eluting with 1:1 ethyl acetate:hexane to give the title 18 compound (IIa) (125 mg, 71%).

19 delta H 5.61 (1H, ddd, J 9.8, 4.5 and 2.7 Hz, 3-H), 21 5.40 (1H, br d, J 9.8 Hz, 5.28 (1H, m, 4.43 22 (1H, m, 3.89 (1H, tt, J 11.2 and 4.5 Hz, 6-H), 23 3.70 (3H, s, C02Me), 2.62 2.59 (2H, m) and 2.51 24 2.48 (2H, d, J 6.6 Hz, 2'-H and 2.46 1.02 (17H, complex multiplets), 1.13 (3H, d, J 7.1 Hz, 26 CO.CHCH 3 0.89 (3H, t, J 7.4 HZ, CH 2

CH

3 and 0.82 (3H, 27 d, 7.0 Hz, CHCH 3 28 29 delta C 209.7, 175.7, 172.1, 132.4, 129.8, 69.4, 66.5, 64.3, 51.7, 48.0, 41.6, 41.5, 41.0, 40.5, 40.4, 39.5, 31 36.5, 35.1, 31.3, 26.5, 21.6, 16.6, 14.7 and 11.5.

32 33 WO 91/15482 PCT/GB9 1/00357 44 1 Step 2 3 Methyl 2S, 4aR, 6R, 8S, 8aS)-6-hydroxy- 4 8-[(2(S)-methyl-l-oxobutyl)oxy]-l, 2, 4a, 5, 6, 7, 8, 8a-octahydro-2-methylnaphthalenyl])-3(R),5(R)-di- 6 hydroxyheptanoate (IIb) 7 8 Triethylborane in THF (1.0 M; 0.66 mmol) was added to a 9 stirred mixture of methanol/THF 6.6 mL) under argon, at room temperature. After 1 hour, the solution 11 was cooled to -70"C and a solution of the ketone (IIa) 12 prepared in step 14 (125 mg, 0.285 mmol) in 13 methanol/THF 6.6 mL) added dropwise. The solution 14 was stirred for 90 minutes then sodium borohydride (13 mg, 0.342 mmol) added in one portion and the solution 16 left at -70*C for 18 hours. Saturated ammonium chloride 17 solution (10 mL) was added, the mixture allowed to warm 18 to room temperature then water added until the solids 19 just dissolved. The aqueous solution was extracted with ethyl acetate (3 x 50 mL) and the combined organics 21 washed with brine (50 mL) and dried. The solvents were 22 evaporated and the residue dissolved in methanol 23 mL), which was warmed, then evaporated again. This 24 process was repeated four times, to leave the title triol of this example (lib) as a gum (125 mg).

26 27 delta H (key peaks) 4.23 (1H, m, 3.89 3.67 28 (2H, m, 6-H and 29 31 32 33 WO 91/15482 PCT/GB91/00357 1 Example 2 2 3 (1S, 2S. 4aR, 6R, 8S. 8aS. 4'R, 6'R. 2 n 2.

4 4a. 5, 6. 7, 8, 8a-octahvdro-6-hydroxy-2-methyl-8-r 2"methyl-1"-oxobutyl) oxy -1-naphthalenyl ethyl 1 tetra- 6 hvdro-4'-hydroxv-2H-pvran-2'-one (I) HO 0 9 11 12 I Me 13 14

HO

16 17 A mixture of the triol of Example 1 (125 mg) and tosic 18 acid monohydrate (20 mg) in dry benzene (15 mL) was 19 stirred for 30 minutes, then the solvent was evaporated under reduced pressure and replaced with fresh dry 21 benzene (15 mL). After stirring for 3.5 hours, the 22 solvent was evaporated and the residue partitioned 23 between ethyl acetate and saturated sodium hydrogen 24 carbonate solution. The organic layer was separated, dried and evaporated. The crude product was 26 recrystallised from ethyl acetate/hexane to give the 27 pure title compound (49.6 mg). The mother liquor was 28 evaporated and the residue crystallised as above, to 29 give a second crop of the title compound (14.6 mg). The two crops were combined, dissolved in ethyl acetate 31 then the solvent allowed to evaporate slowly, giving a 32 total yield of 64.2 mg of lactone.

33 ,0 91/15482 PCT/GB91/00357 46 1 m.pt. 158 159C.

2 3 Found: C, 67.3; H, 8.78%, C 23

H

36 0 6 requires C, 67.6; H 4 8.88%.

6 delta H 5.64 (1H, m, 5.41 (1H, br d, J 9.9 Hz, 7 5.30 (1H, m, 4.60 (1H, m, 4.36 (1H, 8 mi, 3.91 (1H, m, 2.74 (1H, dd, J 17.6 and 9 5.1 Hz, 2'-Hax), 2.61 (1H, m, 2'-Heq), 2.44 1.04 (19H, complex 1.14 (3H, d, J 6.9 Hz, CO.CHCH 3 11 0.91 (3H, t, J 7.4 Hz, CH 2 CH3) and 0.86 (3H, d, J 12 Hz, CHCH3).

13 14 delta C 176.0, 170.3, 132.7, 129.6, 76.0, 69.6, 66.7, 62.5, 41.7, 41.6, 41.0, 39.7, 38.5, 37.2, 35.9, 35.1, 16 32.8, 31.4, 26.6, 23.4, 16.7, 14.7 and 11.6.

17 18 Example 3 19 Sodium 7-(1-r(1S, 2S. 4aR. 6R. 8S. 8aS)-6-hvdroxy- 21 8-r(2(S)-methvl-l-oxobutvl)oxvy-l. 2. 4a. 5, 6. 7. 8, 22 8a-octahydro-2-methvlnaphthalenyll) 5 (R)-di- 23 hydroxyheptanoate (lid) 24 HO CO 2 Na CONa 26 27 0 28 29 Me 31 32 O 33 WO 91/15482 PC'/G B9 1/00357 47 1 A solution of the lactone of example 2 (4.0 mg; 9.8 2 umole) in a mixture of water (50 uL) and sodium 3 hydroxide in methanol (0.1 M; 10.8 umole) was stirred 4 at room temperature for 18 hours, and the solvent evaporated to leave the sodium salt as a gum.

6 7 Example 4 Pharmacology 8 9 IN VITRO DETERMINATION OF INHIBITORY POTENTIAL OF HMG-CoA REDUCTASE INHIBITORS 11 12 HMG-CoA reductase was induced in rats by feeding a 13 normal diet supplemented with 3% cholestyramine resin 14 for one week prior to sacrifice. The livers were excised from the sacrificed rats and microsomal pellets 16 prepared by the method of Kleinsek et al, Proc. Natl.

17 Acad. Sci. USA, 74 pp 1431-1435, 1977. Briefly, 18 the livers were immediately placed in ice-cold buffer I 19 and homogenised in a Potter-Elvehjem type glass/teflon homogeniser (10 passes at 1000 rpm). The homogenate was 21 centrifuged twice at 20,000 x g to remove debris. The 22 supernatant was centrifuged at 100,000 x g for 23 minutes, the microsomal pellet resuspended in buffer II 24 and centrifuged at 100,000 x g for 75 minutes. The resultant pellet was stored at -70'C until required for 26 assay purposes.

27 28 Buffer I Buffer II 29 50 mM KPO 4 pH 7.0 50 mM KPO 4 pH 31 0.2 M sucrose 0.2 M sucrose 32 2 mM DTT 2 mM DTT 33 50 mM EDTA WO 91/15482 PCT/GB91/00357 48 1 2 Assay of HMG-CoA Reductase Activity and Determination 3 of Activity of Inhibitors 4 Membrane bound enzyme isolated as above is used for 6 determining the activity of inhibitors. The assay is 7 performed in a total volume of 300 gL in 100 mM KPO 4 pH 8 7.2 buffer, containing 3 mM MgC12, 5 mM 9 glucose-6-phosphate, 10 mM reduced glutathione, 1 mM NADP, 1 unit glucose-6-phosphate dehydrogenase, and 1 11 mg/mL BSA, with resuspended enzyme. Putative inhibitors 12 are dissolved in dimethylsulphoxide and 10 ML aliquots 13 added to the incubation.

14 The assay is pre-incubated at 37°C for 10 minutes and 16 initiated by the addition of 0.1 pCi 3-hydroxy- 17 3-methyl-[3- 14 C]glutaryl coenzyme A (52 Ci/Mole) 18 followed by incubating the complete reaction at 37*C 19 for 10 minutes. At the end of this period the reaction is stopped by adding 300 gL of a 10 mM mevalonolactone 21 solution in 0.1 M hydrochloric acid and the mevalonic 22 acid product allowed to lactonise for a further period 23 of 30 minutes. The product is then isolated by 24 chromatography using BIO-REX 5 resin and the enzyme activity quantified by liquid scintillation 26 spectrophotometry. (The expression "BIO-REX" is a trade 27 mark.) 28 29 Appropriate controls are included in the assay and IC 50 values obtained by graphical means. The compound of 31 Example 3 exhibited an IC 50 of 14 nM by this method, 32 which compared favourably with the corresponding value 33 for the known compound dihydromevinolin (30 nM).

WO 91/15482 PCT/GB91/00357 49 1 2 Examples of unit dosage compositions are as follows: 3 4 Example Capsules: 6 Per 10,000 7 Ingredients Per Capsule Capsules 8 9 1. Active ingredient (Cpd of Formula I) 40.0 mg 400 g 11 2. Lactose 150.0 mg 1500 g 12 3. Magnesium 13 stearate 4.0 mq 40 g 14 194.0 mg 1940 g 16 Procedure for capsules: 17 18 Step 1. Blend ingredients No. 1 and No. 2 in a 19 suitable blender.

Step 2. Pass blend from Step 1 through a No. 30 mesh 21 (0.59 mm) screen.

22 Step 3. Place screened blend from Step 2 in a 23 suitable blender with ingredient No. 3 and 24 blend until the mixture is lubricated.

Step 4. Fill into No. 1 hard gelatin capsule shells 26 on a capsule machine.

27 28 29 31 32 33 091/15482 PCT/GB91/00357 1 Example 6 2 3 Tablets: 4 Per 10,000 Ingredients Per Tablet Tablets 6 7 1. Active ingredient 8 (Cpd of Form. I) 40.0 mg 400 g 9 2. Corn Starch 20.0 mg 200 g 3. Alginic acid 20.0 mg 200 g 11 4. Sodium alginate 20.0 mg 200 g 12 5. Magnesium 13 stearate 1.3 mq 13 g 14 101.3 mg 1013 g 16 17 Procedure for tablets: 18 Step 1. Blend ingredients No. 1, No. 2, No. 3 and No.

19 4 in a suitable mixer/blender.

Step 2. Add sufficient water portionwise to the blend 21 from Step 1 with careful mixing after each 22 addition. Such additions of water and mixing 23 until the mass is of a consistency to permit 24 its conversion to wet granules.

Step 3. The wet mass is converted to granules by 26 passing it through an oscillating granulator 27 using a No. 8 mesh (2.38mm) screen.

28 Step 4. The wet granules are then dried in an oven at 29 140°F (60 0 C) until dry.

Step 5. The dry granules are lubricated with 31 ingredient No. 32 Step 6. The lubricated granules are compressed on a 33 suitable tablet press.

WO 91/15482 PCT/GB91/00357 1 2 Example 7 3 4 Intramuscular Injection: Ingredient 6 1. Formula I compound 7 Active ingredient 8 2. Istonic buffer 9 solution pH 4.0.

Per ml. Par litre 10.0 mg 10 g q. s.

q.s.

Procedure: Step 1. Dissolve the active ingredient in the buffer solution.

Step 2. Aseptically filter the solution from Step 1.

Step 3. The sterile solution is now aseptically filled into sterile ampoules.

Step 4. The ampoules are sealed under aspetic conditions.

Example 8 Suppositories: Per 1,000 Supp Ingredients 1. Formula I compound Active ingredient 2. Polyethylene Glycol 1000 3. Polyethylene Glycol 4000 Procedure: Step 1. Melt ingredient No.

Per Supp.

40.0 mg 40 g 1350.0 mg 450.0 mq 1840.0 mg 1,350 g 450 q 1,840 g 2 and No. 3 together and WO 91/15482 PCr'/GB1/00357 52 1 stir until uniform.

2 Step 2. Dissolve ingredient No. 1 in the molten mass 3 from Step 1 and stir until uniform.

4 Step 3. Pour the molten mass from Step 2 into suppository moulds and chill.

6 Step 4. Remove the suppositories from moulds and 7 wrap.

8 9 11 12 13 14 16 17 18 19 21 22 23 24 26 27 28 29 31 32 33

Claims (10)

1. A compound of either general formulae I and II: HO 0 HO. R 3 represents a hydrogen atom or a substituent R 4 or M; R 4 represents a C1- 5 alkyl group, or a C1-5 alkyl group substituted with a group chosen from substituted phenyl, dimethylamino, or acetylamino; R 5 represents a hydrogen atom or C 1 -3 alkyl group; M represents a cation capable of forming a pharmaceutically acceptable salt; Q represents C=O or CHOH; and a is a double bond and b is a single or double bond, or a pharmaceutically acceptable salt thereof. PCT/PR 9 1 OP n 57 March 1992 54 20 03 9?2 1

2. A compound claimed in claim 1, which has one or 2 more of the following substituents independently or in 3 any combination: 4 R 1 represents C 4 6 branched alkyl; 6 R 3 is R 4 7 R 4 represents C 1 5 alky.; 8 Q represents CHOH; and/or 9 b is a single bond. 11

3. A compound according to claim 2, wherein R 1 12 represents a branched C 4 alkyl group and R5 represents 13 methyl. 14

4. (1S, 2S, 4aR, 6R, 8S, 8aS, 41R, 61R, 16 2, 4a, 5, 6, 7, 8, 8a-octahydro- 17 6-hydroxy-2-methyl-8-[ methyl-l"1-oxobutyl) oxy]-l- 18 naphthalenyl] ethyl] tetra- hydro-4' -hydroxy-2H-pyran- 19 2'-one 21

5. A compound according to claim 2, wherein R 1 22 represents a branched C 4 alkyl group, R 3 represents 23 methyl or ethyl, R 5 represents methyl, v~nd Q represents 24 the group CHOH. 26

6. Methyl 7-(l-E(lS, 2S, 4aR, 6R, 8S, 8aS)-6- 27 hydroxy-8-[(2(S)-methyl-l-oxobutyl)oxy]-l, 2, 4a, 28 6, 7, 8, 8a-octahydro-2-methylnaphthalenyl] 1-3(R), 29 5(R) -dihydroxyheptanoate 31 A cempeu-nd as dofir.ed In any cne ef the elaims 1 32 to6 -fer,s in ffled-L-.- 33 I, ~'atcn( Office U TTT H T 55

7. A method of treatment of hypercholesterolaemia, hyperlipoproteinaemia, or atherosclerosis, comprising the step of administering to a mammal in need of such treatment an effective amount of a compound as defined in any one of Claims 1 to 6, or a pharz ceutically acceptable salt thereof.

8. A pharmaceutical composition comprising a compound as defined in any of claims 1 to 5, or a mixture of such compounds, and a pharmaceutically acceptable carrier thereof.

9. A composition as claimed in claim 8 comprising a pharmaceutically acceptable non-toxic cationic polymer capable of binding bile acids in a non-reabsorbable form in the gastrointestinal tract.

10. A process for the preparation of a compound of either general formulae I or II as defined in any one of claims 1 to 6, the process comprising the steps of: deprotecting and optionally reducing a compound of general formula XIV to form a compound of general formula IIa; and optionally converting a compound of general formula I or IIa directly or indirectly into another compound of general formula I or II. DATED this 16th day of December 1994 BRITISH BIOTECHNOLOGY LIMITED By Their Patent Attorneys: GRIFFITH HACK CO Fellows Institute of Patent Attorneys of Australia