CA1171424A - ( )-(4r, 6s) - (e)-6-[2-(4'fluro-3,3', 5-trimethyl- [l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6- tetrahydro-4-hydroxy-2h-pyran-2-one - Google Patents

Abstract

TITLE OF THE INVENTION
(+)-(4R, 6S)-(E)-6-[2-(4'-fluoro-3,3',5-trimethyl-[l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one ABSTRACT OF THE DISCLOSURE
(+)-(4R, 6S)-(E)-6-[2-(4'-fluoro-3,3',5-trimethyl-[l,1'-biphenyl]-2-yl)ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one is a very potent inhibitor of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A
reductase, a major rate limiting enzyme in cholesterol biosynthesis.

Description

~1 ~'7~ 4 TITLE OF THE INVENTION
(+) (4R, 6S)-(E)-6-[2-(4'-fluoro-3,3',5-trimethyl-[l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one SUMMARY OF THE INVENTION
: This invention relates to a hypocholesterol-emic and hypolipemic compound which is a member of the class of compounds having the structure (I) : HO ~ O
~=/
:
Rl 5 ~ 2 ~ ~ ' ::
3~' '^' ~ :
.

~ 2 - 16643 and the corresponding dihydroxy acid resulting from the hydrolytic opening of the lactone ring, and pharmaceutically acceptable salts of said acid.
Specifically, it is the compound of structure I
wherein E is trans-CH=CH-, Rl is 6-(4'-fluoro-3'-methyl-l-phenyl), R2 is 2-methyl, and R3 is 4-methyl.

BACKGROUND OF THE INVENTION
Recently, Endo et al, reported (U.S. Letters Patent 4,049,495, Patent 4,137,322 and Patent 3,983,140) the production of a fermentation product which was quite active in the inhibition of cholesterol biosynthesis. This natural product, now called compac-tin, was reported by Brown et al., (J.
Chem. Soc. Perkin I, 1165 (1976)~ to have a complex mevalonolactone structure.
Recent U.S. Patents 4,138,425 and 4,255,444 disclose a group of synthetic compounds of the generic formula II A

HO

Rl t ~ R2 R

in which A is -CH3 and hydrogen respectively, E
represents a direct bond, a C1 3 alkylene bridge or a vinylene bridge and the R groups represent a variety of substituents.

. ,`.~

A patent application of Merck & Co., Inc., E.P. publication 0 024 348, discloses a similar series of synthetic compounds in which A is hydrogen and in which the lactone ring :has a (4R)-trans configuration, the isomer which demonstrates all of the antihypercholesterolemic activity of the unresolved, cis-trans lactone mixture.
Now, with the present invention there is provided the compound, (+)-(4R, 6S)-(E)-6-[2-(3,3',5-trimethyl-4'~fluoro-[l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one, which has an unexpectedly high order of efficacy relative to other members of the genus of which it is a member and which is disclosed in the above-mentioned E.P.
publication 0 024 348.

DETAILED DESCRIPTION OF THE INVENTION
This invention relates to a hypocholesterol-emic and hypolipemic compound having the structure 20 III. H
HO ~ o CH3 ~

~ ~ ~ 3 : CH3 III
~: 30 _ 4 _ 16643 or the corresponding dihydroxy acid resulting from the hydrolytic opening of the lactone ring, or a pharmaceutically acceptable salt of the acid, the compound being the enantiomer with a 4 (R) configuration in the tetrahydropyran moiety of the trans racemate shown in the structures I and III.
The designation 4 (R) with respect to this compound indicates that the absolute configuration in space at the 4-carbon of the tetrahydropyranone ring is believed to be the Rectus (R) series, and it is dextrorotatory, and it is an unexpectedly potent inhibitor of cholesterol biosynthesis, surpassing the inhibitory activity of compactin.
The cited U.S. Patents 4,138,425 and 4,255,444 show no recognition of the stereochemistry of compounds II, let alone the fact that an unexpectedly large improvement in the activity would result from the separation of the cls and trans racemates and the latter's resolution, especially when the preferred 2,4,6-trisubstitution occurs in the phenyl ring. However, E.P. publication 0 024 348 discloses that the 4 (R) enantiomers of the trans racemates corresponding to formula I specifically inhibit with high potency the activity of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, which is known to be the enzyme involved in the rate limiting step in the process of cholesterol biosynthesis.
The compound III of this invention is the most efficacious member of the latter series. The inhibitory activity of these compounds for the biosynthesis of cholesterol has been measured by two '7~ ~4 _ 5 _ ~6643 methods. The experimental method A of U.S. 4,138,425 was the ln vitro method of H. J. Knauss, et al., J.
Biol. Chem., 234, 2835 (1959) and the activity was expressed as the molar concentration IC50(M) necessary for the inhibition of 50% of the enzymatic activity. The experimental method B of U.S.
4,198,425 was the method of A. A. Kandutsch, et alO, J. Biol. Chem., 248, 8403 (1973) for measuring 14C-cholesterol biosynthesis from acetic acid-14C
in mouse L cells. The activity is expressed for inhibition of 50% of the biosynthesis of cholesterol.
The results obtained in these two asays for the compounds of Structure II, as reported in the cited U.S. patents, show IC50 values of 10 4 to 10 6 in both tests. The smallest 50% effective dose cited is about 4 x 10 6, whereas the value for compactin, in the same tests, is about 0.8 x 10 8.
E.P. publication 0 024 348 shows that the inhibitory potency is greatly increased by separation of isomers especially when this is combined with optimal selection of a 2,4,6-arrangement of Rl, R2 and R3 in the phenyl ring of structure II and especially when A is hydrogen and E is trans-CH=CH-.
Thus the (+) trans enantiomer of 6-E2-(2f4-dichloro-6-(phenylmethoxy)phenyl)ethyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one gives an IC50 of 6.8 x 10 8 in the test by method A. An even more potent compound is the (+) trans enantiomer of (E)-6-[2-(3,5-dichloro-4'-fluoro[l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one and the compoun~ of the present invention, the (~)-trans enantiomer of (E)-6-[2-(4'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one, is several times more potent.
The compounds were tested as the sodium salts of their corresponding dihydroxy acld forms.
The compound of this invention has the added advantage of having no chloro substituents. Although the metabolism of the compound is not completely known it is advantageous not to have a compound that could be metabolized to a polychlorobiphenyl (PCB), a class of substances known to be involved in carcinogenesis.
The Flow Sheet shows the synthesis of the 4 (R) trans lactone of this invention. The number of each step in the Flow Sheet corresponds with the number of each reaction description and each step of Example 1 which follow. Steps 5 through 3 represent the novel process of this invention.

1"~

CHO N;~

[~ CH3 S t 1 ~ C 3 ~ Step 3 3 ~CN3 _ 2 ~ Step 4 ~; ` /

~: .
~ : ' : ~ ., : :`

::
'. : .

- , ~C02C~3 ~CH ~ ~ CH3 ~¦~Step 6 HO~--`Co2CH3 HO~a C 3~ OH CH3 ? Step 7 3, F~ ~ S~8 '3 ~3fU3 CH3~ Step 9 C' ~ -I I I

.' ' ' ' ~ ' ' ': ' ' ` `

a.~

_ 9 _ 16643 REACTIONS IN THE E`LOW SHEET

1. Reaction with ani:Line in refluxing toluene.

2. Reaction with Pal:Ladium (II) acetate in acetic acid at reflux.

3. Reaction with a substituted Grignard 10 reagent CH3 F - ~ MgBr in a suitable solvent such as benzene or toluene in the presence of triphenylphoshine followed by hydrolysis with 6N HCl at ambient temperature.

4. Aldol Reaction.

a) The classical Aldol synthesis in which acetaldehyde is condensed with the starting benzaldehyde, the resulting ~-hydroxyaldehyde is acetylated with acetic anhydride and acetic acid is eliminated thermally to give the corresponding cinnamaldehyde.
b) The directed Aldol condensation in which the anion of an appropriately N-substituted ethylidenylimine, such as ethylidenecyclohexylimine and the like, is condensed with the starting benzaldehyde at or below room temperature in an aprotic solvent, such as THF and the like, to afford a ~-hydroxy ~-phenyl-propylidenylimine which, upon concomitant dehydration and imine hydrolysis in an acidic medium, such as dilute aqueous HCl, provides the corresponding cinnamaldehyde.
c) The use of a nucleophilic acetaldehyde equivalent in which cis-2-ethoxyvinylithium, generated from cls-l-ethoxy-2-tri-n-butylstannyl-ethylene, is condensed with the starting benzaldehydeto give an allylic alcohol which is subse~uently rearranged, under suitable acidic conditions, to the corresponding cinnamaldehyde.

5. Dianion Step. Reaction with the dianion of acetoacetic ester in a suitable aprotic solvent such as THF, dioxane and the like.

6. Reduction with NaBH4 in a suitable solvent such as methanol, ethanol and the like at or below room temperature.

7. Lactonization. Saponification by base (e.g. NaOH) in aqueous alcohol followed by acidification and cyclodehydration by heating in toluene followed by separation of the cis and trans mixture by chromatogxaphy on silica gel or crystallization.

8. Resolution of the trans racemate into its enantiomers by treating the (+)-trans lactone with either d-(+) or l-(~ -methylbenzylamine to give the diastereomeric dihydroxy amides which are separated by chromatography or crystallization.

:

~ 16643 3. Hydrolysis of each pure diastereomeric amide under basic conditions, such as ethanolic NaO~
and the like to afford the corresponding enantiomer-ically pure dihydroxy acid which, upon lactonization, e.g., in refluxing toluene, provides the pure (+)-trans enantiomer. Stereochernistry depends on the absolute stereochemistry of the diastereomeric amide from which it is derived.

A further aspect of the present invention is a pharmaceutical composition consisting of compound III in association with a pharmaceutical vehicle or diluent. The pharmaceutical composition can be formuated in a classical manner utilizing solid or liquid vehicles or diluents and pharmaceutical additives of a type appropriate to the mode of desired administration. The compounds can be administered by an oral route, for example, in the form of tablets, capsules, granules or powders, or they can be administered by a parent~ral route in the form of injectable preparations. The dose to be administered depends on the unitary dose, the symptoms, and the age and the body weight of the patient. A dose for adults is preferably between 200 and 2,000 mg per day, which can be administered in a single dose or in the form of individual doses from 1-4 times per day.
A typical capsule for oral administration contains active ingredient (250 mg), lactose (75 mg) and magnesium stearate (15 mg). The mixture is passed through a 60 mesh sieve and packed into a No. 1 gelatin capsule.

A typical injectible preparation is pr~duced by asceptically placing 250 mg of sterile active ingredient into a vial, asceptically freeze-drying and sealing. For use, the contents of the vial are mixed with 2 ml of physiological saline, to produce an injectible preparation.
The compound of this invention also has useful antifungal activity. For example, it may be used to control stralns of Penicilium sp., Aspergillus niger, Cladosporium sp. ! Cochliobolus miyabeonus and Hilminthosporium cynodnotis. For this utility it is admixed with suitable formulating agents, powders, emulsifying agents or solvents such as aqueous ethanol and sprayed or dusted on the plants to be protected.
This invention can be illustrated by the following examples in which ratios of solvents are in volumes and percentages, unless otherwise indicated, or by weight.

EXAMPLE
(+)-(4R,6S)-(E)-6-[2-(4'-fluoro-3,3',5~trimethyl[1,1'-biphenyl]-2-yl)-ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one Step 1: Preparation of N-[(2,4-Dimethylphenyl)-methylene]benzeneamine A mixture of 2,4-dimethylbenzaldehyde (53.7 g, 0.4 mole), freshly distilled aniline (37.5 g, 0.4 mole) and toluene (150 ml) was heated at reflux under 0 a Dean-Stark trap for 2 hours. The solution was cooled and then concentrated under reduced pressure.
Distillation of the residue at 0.2 mm gave the product as a yellow oil (81.3 g, 97~ yield), bp 122-130, which solidified on cooling.
Step 2: Preparation of Bis[~-(Acetato-O:O')bis(3,5-dimethyl-2-[(phenylimino)methyl]phenyl-C,N]-dipalladium A mixture of N-[(2,4-dimethylphenyl)-methylene]benzeneamine (58.7 g, 0.28 mole) and palladium (II) acetate (62.9 g, 0.28 mole) in acetic acid (1 L) was hea-ted at reflux with stirring for one hour. After cooling (to 50), the reaction mixture was filtered (gravity) and the filtrate poured into 5 water (4 L). The aqueous mixture was stirred overnight and then filtered. The collected solid was washed with several portions of cold water, air dried for several hours (with suction) and dried in a vacuum oven ( 45) for 2 days to afford the desired product as a yellow-orange solid (101.3 g, 97~ yield).

'7 ~ /~2~

- 1~ - 16643 Step 3: Preparation of 4'-Fluoro-3,3',5-trimethyl-1,1'-biphenyl-2-carboxaldehyde In a 1 L, 3-neck flask equipped with a magnetic stirring bar, N2 inlet tube~ dropping funnel (250 ml), and reflux condenser capped with a drying tube, was placed Mg turnings 4.5 g (0.185 mole). This equipment was heated with a heat gun under N2 and then allowed to cool to room temperature. 5-Bromo-2-fluorotoluene (35.0 g, 0.185 mole) was dissolved in 300 ml of ether and 40 ml of this solution was added to the Mg turnings. After mild heating with a heat gun to initiate the reaction, the remainder of the ether solution was added at such a rate as to maintain reflux ( one hour). The mixture was stirred at reflux for an additional 30 minutes and cooled to room temperature.
Meanwhile, the Pd complex (45.2 g, 0.06 mole) from Step 2 had been added to 1 L of toluene (3 L, 3-neck flask). The resulting mixture was stirred vigorously and heated at reflux for one hour using a Dean-Stark trap to collect traces of water.
The mixture was cooled to room temperature under N2, triphenylphosphine (64.0 g, 0.24 mole) was added and the mixture was stirred for 30 minutes.
The Grignard reagent was added b~ means of a dropping funnel in a slow, steady stream and the reaction mixture stirred for one hour. After addition of 200 ml 6N HC1, the mixture was stirred for an additional one hour and filtered. The collected solid was washed with several portions of toluene. The filtrate and washings were combined and separated.
The toluene ether solution was washed with 2 x 200 ml '7~Z~

of brine and dried over MgSO4. Filtration and evaporation yielded a black oil ( 70 g) which was chromatographed using a 120 mm "Still" col~mn (1.5 kg of 230-400 mesh Silica Gel) and 40~ (v:v) CH2C12-hexane. There were collected 2 fractions o~ 1000 mland 20 fractions of 500 ml, the pure product being ~ound in fractions 5-14. Fractions 14-17 contained a mixture of the desired aldehyde plus 2,4-dimethyl-benzaldehyde. There was obtained 19-21 g of combined product as a pale yellow solid ( 70~ yield); one spot on tlc (Rf=0.30 on Silica Gel GF plate with 40% (v:v) CH2C12-hexane), mp 75-78C. The mp of pure sublimed product is 78-80.

Step 4: Preparation of 3-(4'-Fluoro-3,3',5-trimethyl-[l,l'~biphenyl]-2-yl)-2-~ro~enal A dry 1 L, 4-neck flask equipped with a magnetic stirring bar, N2 inlet tube, thermometer, addition funnel (capped with a drying tube) and septum was charged with a solution of (Z)-l-ethoxy-2-tributylstannylethylene (68.5 g, 0.19 mole) in dry THF (200 ml3 and then cooled in a dry ice-acetone bath. n-Butyllithium (1.48 M in hexane, 130 ml, 0.19 mole) was added via syringe over a period of 25 minutes. The resulting mixture was stirred at -75 under N2 for one hour. A solution of 4'-fluoro-3,3',5-trimethyl-1,1'-biphenyl-2-carboxaldehyde (40.7 g, 0.168 mole) in THF (150 ml) was added dropwise ( one hour) at -75. After stirring for 10 minutes, the cooling bath was removed and the reaction mixture was stirred at ambient temperature for 1-1/2 hours.
Saturated acl. NaHCO3 (150 ml) was added in a slow 2~

steady stream and the mixture, distributed between ether (500 ml) and water (500 ml). After separating the layers, the aqueous phase was extracted with ether (2x). The ether extracts were combined, washed with cold water and brine, dried over MgSO4, filtered and evaporated.
The residual oil was taken up in THF (200 ml) and the solution treated with 6N HCl (25 ml) and stirred at room temperature for one hour. The mixture was diluted with cold water and extracted (3x) with ether. The ether extracts were combined, washed with cold water and brine, and dried over MgSO4. Filtration and evaporation yielded a yellow brown oil ( 120 g), which was chromatographed using 2.6 kg of Silica Gel (230-400 mesh) in a 140 mm "Still" column. Elution with CH2C12-hexane t3:1, v:v) gave 21 fractions (1 L each), the major portion of product being located in fractions 11-19. After evaporation, the oil was taken up in warm hexane (60 ml); the solution was seeded and cooled at room temperature to give a white crystalline solid (20.6 g), mp 82-85. Fractions 9 and 10 were handled separately and provided after recrystallization from hexane an additional 3.6 g of product; total quantity was 24.2 g (54~ yield).

Step 5: Preparation of Methyl (E)-7-(4'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl) 5-hydroxy-3-oxo-6-heptenoate Methyl acetoacetate (12.31 g~ 0.106 mole) was added dropwise to a stirred suspension of sodium hydride (50~ oil suspension)(5.09 g, 0.106 mole) in l .a, ~

dry THF (180 ml) at 0 (internal, keeping temperature 10C) and under N2. The resulting solution was stirred at 0 for 15 minutes and then treated with a solution of 1.40 M n-butyllithium in hexane (78.7 ml, 0.106 mole) added dropwise ( 20 minutes) via syringe. After stirring at 0 for 20 minutes, the bath was replaced by an ice-acetone bath and stirred for an additional 5 minutes. A solution of 3-(4'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)-2-propenal (26.5 g, 0.099 mole) in dry THF (150 ml) was added dropwise ( 15 minutes) at such a rate as to keep the temperature 10C. The reaction mixture was stirred at 0C ~or 30 minutes and then quenched by the slow addition of 6N HCl (45 ml). The mixture was then diluted with water (300 ml) and extracted (3x) with ether. The ether extracts were combined, washed with cold water and brine, dried (MgSO~), filtered and evaporated leaving a yellow oil ( 38 g); one major spot on tlc (Silica Gel GF) at Rf = 0.43 ~5% (v:v) acetone-CH2C12).

Step 6: Preparation of Methyl (E)-7-(4'-Fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenoate Sodium tetrahydridoborate (1.92 g, 0.0507 mole) was added portionwise over a period of 10 minutes and with stirring to a cold solution (0C) of methyl (E)-7-(4'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)-5-hydroxy-3-oxo-6-hep-tenoate (37.5 g) in methanol (400 ml). The clear reaction mixture was stirred at 0C for 15 minutes and then diluted with water (300 ml). While maintaining the temperature .'71.~4 below 10C with an ice-bath, the mixture was acidified with 12N HCl ( 22 ml). Following extrac-tion with ether (3 x 250 ml), the ether extracts were combined, washed with cold water and brine, dried (MgSO4) and evaporated to a viscous oil ( 38 g).
The product showed one major spot on tlc (silica gel GF) with Rf 0.21 (10% (v:v) acetone-CH2C12).

Step 7: Preparation of (+) trans (E)-6-[2-(4l-Fluoro--3,3',5-trimethyl[l,l'-biphenyl]-2-yl)-ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one _ _ A solution of methyl (E)-7-(3'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)-3,5-dihydroxy-6-h eptenoate ( 37.7 g, .0976 mole) and lN sodium hydroxide (lO0 ml, 0.10 mole) in methanol (150 ml) was stirred at room temperature for 15 minutes.
After removal of the methanol by evaporation ( 30), the reaction mixture was diluted with water (600 ml), acidified with conc. HCl and extracted with ether (3 x 300 ml). The ether extracts were combined, washed with cold water and brine, dried over MgSO~, filtered, and evaporated to yield a yellow-orange oil ( 37 g)-A solution of the crude oily diol acid in toluene (300 ml) was heated at reflux under a Dean-Stark trap for one hour. Evaporation provided a yellow oil ( 37 g) which was a mixture of cis and trans lactones. The crude product was chromatographed using a 140 mm "Still" column (230-400 mesh Silica Gel) and 10% (v:v) acetone-CH2C12. After - . , collecting 3 fractions of 1 L and 25 fractions of 500 ml, elution was continued with 20% (v:v) acetone-CH2C12 collecting 6 fractions of 1 L each, containing primarily cis racemate ( 84%). Fractions 12-15 of the 500 ml cuts were combined and evaporated to give the trans racemate as a pale yellow oil ( 90%
pure by HPLC). This was crystallized from Et2O-hexane to give a solid (5 g) mp 115-117.
Fractions 16-25 were combined and evaporated to give a pale yellow oil ( 12 g), which was a (6:4) mixture of trans and cls racemates.
This mixture ( 12 g) was re-chromatographed in two runs with a Waters Prep LC500, employing 2 prep PAK-500 silica cartridges in series and eluting 15 with acetone-CH2C12 (1:9, v:v). Using the shave recycle technique, the trans isomer (5 g) and the cls racemate (4 g) were obtained. The samples of the trans racemate, collected from the two chromatographic separations, were combined to give a 20 white crystalline solid (10 g) mp 115-117. All cis fractions from chromatography and the Waters separation were combined to give 9 g which crystallized on stand1ng.

Step 8: Preparation of (3R,5S)-N-((S)-~-Methyl-benzyl)-7-(4'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenamide A solution of ~+)-trans-(E)-6-[2-(4'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6 30 tetrahydro-4-hydroxy-2H-pyran-2-one (10 g, 28.2 mole) and (S)-(~ -methylbenzylamine (36.35 ml, 34.17 g, 282 mmole) in THF (35 mI) was stirred at gentle :
. . . .

reflux under N2 for 16 h. The reaction mixture was then cooled, diluted wit Et2O (400 ml) and successively washed with H2O (2 x 200 ml), 3N HCl (2 x 300 ml) and brine (2 x 200 ml), dried (MgSO4) and filtered. The filtrate was evaporated leaving the mixture of intermediate d:iastereomeric amides as a pale amber gum.
The residual gum was digested in Et2O (100 ml) and treated with hexane until turbid. This was seeded and cooled at 0. Solid, 1.5 g, m.pO 110-12, was collected by filtration. The filtrate was evaporated to a gum ( 12 g).
This mixture of diastereomeric amides was chromatographed using a Waters Prep LC500. Separa-tion of the mixture was accomplished using two prepPAX-500/silica cartridges in series eluting with methylene chloride-acetone (80:20, v:v). The mixture was divided and run in two separate elutions. Using the shave recycle technique, the two diastereoisomers (1) (4.5 g, m.p. 110-12) and (2) (3.5 g, 52.6~ of theory) m.p. 86-9 were obtainedO Compound (2) which was 99~ pure by ~P~C is the desired diastereoisomer.
Mother liquors from recrystallization of (2) were combined (1.9 g, 85~ (2)).
Ste~_9: Preparation of (+)-(4R,6S)-(E)-6-[2-(4'-Fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)-ethenyl}-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one A solution of N-((S)-~-methylbenzyl)-7-(4'-fluoro-3,3'J5-trimethyl[l,l'-biphenyl]-2-yl)-3,5-di-~1~7~L'Z~L

- 21 - 166~3 hydroxy-6-heptenamide (3.5 g, 7.35 mmol), diastereoisomer (2) in step 8 above, in EtOH (300 ml) containing lN NaOH (44 ml, 44 mmole) and H2O (44 ml) was refluxed under N2 for 14 h. The solvent was removed ln vacuo ( 40). The residue was dissolved in ice water (300 ml) and Et2O (500 ml), then cooled and stirred at 0 while 3N HCl (50 ml) was slowly added. The organic layer was separated and washed successively with ice-cold lN HCl (200 ml) 10 and brine (2 x 200 ml), dried (MgSO4) and filtered. After evaporation of solvent ln vacuo, the remaining residue was dissolved in toluene (500 ml) and refluxed under a Dean-Stark trap for 2 h.
Evaporation provided a yellow oil which was 15 chromatographed on a 50 mm "Still" column (230-400 mesh Silica Gel) using acetone-methylene chloride (10:30, v:v) and collecting 10 ml fractions. The product was found in cuts 19-31 followed by cuts slightly contaminated with cis lactone. Less than pure fractions were combined and, after evaporation, further enriched by crystallization from Et2O-hexane. The combined yield was 1.6 g, 61%. This was 99.99% pure by HPLC, m.p. ~7~9 E~]D5 + 40.56 (C = 1.075, CHC13).

.

Claims (2)

WHAT IS CLAIMED IS:

1. A process for the preparation of (+)-(4R,6S)-(E) -6-[2-(4'-fluoro-3,3',5-trimethyl[l,l'-biphenyl]-2-yl)ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one characterized in that:
a) condensation of the dianion of acetoacetic ester with an aldehyde of structure:
to give structure:

b) reduction of the keto group to an hydroxyl;
c) saponification of the ester, lactonization and separation of the cis and trans racemates;
d) formation of the diastereomeric .alpha.-methyl benzylamides of the trans-racemate and separation of the diastereomers; and e) hydrolysis of the diastereomer containing the product (+)-trans enantiomer and lactonization.

2. The compound, (+)-(4R,6S)-(E)-6-[2-(4'-fluoro-3,3',5-trimethyl[1,1'-hiphenyl]-2-yl)-ethenyl]-3,4,5,6-tetrahydro-4-hydroxy-2H-pyran-2-one, or a pharmaceutically acceptable salt thereof, when prepared by the process defined in Claim 1 or by an obvious chemical equivalent.