(+)-(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, a process for preparing and a pharmaceutical composition containing the same

Claim 2

Reaction with aniline in refluxing toluene.

Claim 3

Reaction with Palladium (II) acetate in acetic acid at reflux.

Claim 4

Reaction with Palladium (II) acetate in acetic acid at reflux.

Claim 5

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

Claim 6

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

Claim 7

Aldol Reaction. a) The classical Aldol synthesis in which acetaldehyde is condensed with the starting benzaldehyde, the resulting 6-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 β-hydroxy-β-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 cis-l-ethoxy-2-tri-n-butylstannyl- ethylene, is condensed with the starting benzaldehyde to give an allylic alcohol which is subsequently rearranged, under suitable acidic conditions, to the corresponding cinnamaldehyde.

Claim 8

Aldol Reaction. a) The classical Aldol synthesis in which acetaldehyde is condensed with the starting benzaldehyde, the resulting 6-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 β-hydroxy-β-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 cis-l-ethoxy-2-tri-n-butylstannyl- ethylene, is condensed with the starting benzaldehyde to give an allylic alcohol which is subsequently rearranged, under suitable acidic conditions, to the corresponding cinnamaldehyde.

Claim 9

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

Claim 10

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

Claim 11

Reduction with NaBH 4 in a suitable solvent such as methanol, ethanol and the like at or below room temperature.

Claim 12

Reduction with NaBH 4 in a suitable solvent such as methanol, ethanol and the like at or below room temperature.

Claim 13

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 chromatography on silica gel or crystallization.

Claim 14

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 chromatography on silica gel or crystallization.

Claim 15

Resolution of the trans racemate into its enantiomers by treating the (+)-trans lactone with either d-(+) or 1-(-)-a-methylbenzylamine to give the diastereomeric dihydroxy amides which are separated by chromatography or crystallization.

Claim 16

Resolution of the trans racemate into its enantiomers by treating the (+)-trans lactone with either d-(+) or 1-(-)-a-methylbenzylamine to give the diastereomeric dihydroxy amides which are separated by chromatography or crystallization.

Claim 17

Hydrolysis of each pure diastereomeric amide under basic conditions, such as ethanolic NaOH and the like to afford the corresponding enantiomerically pure dihydroxy acid which, upon lactonization, e.g., in refluxing toluene, provides the pure (+)-trans enantiomer. Stereochemistry 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 parenteral 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 produced 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 strains 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.

Claim 18

Hydrolysis of each pure diastereomeric amide under basic conditions, such as ethanolic NaOH and the like to afford the corresponding enantiomerically pure dihydroxy acid which, upon lactonization, e.g., in refluxing toluene, provides the pure (+)-trans enantiomer. Stereochemistry 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 parenteral 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 produced 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 strains 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.

Claim 19

EXAMPLE

Claim 20

EXAMPLE

Claim 21

(+)-(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

Claim 22

(+)-(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

Claim 23

Step 1: Preparation of N-[(2,4-Dimethylphenyl)-methylene]benzeneamine

Claim 24

Step 1: Preparation of N-[(2,4-Dimethylphenyl)-methylene]benzeneamine

Claim 25

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 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.

Claim 26

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 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.

Claim 27

Step 2: Preparation of Bis[p-(Acetato-0:0')bis(3,5-dimethyl-2-[(phenylimino)methyl]phenyl-C,N]-dipalladium

Claim 28

Step 2: Preparation of Bis[p-(Acetato-0:0')bis(3,5-dimethyl-2-[(phenylimino)methyl]phenyl-C,N]-dipalladium

Claim 29

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 heated at reflux with stirring for one hour. After cooling (to 50°), the reaction mixture was filtered (gravity) and the filtrate poured into 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).

Claim 30

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 heated at reflux with stirring for one hour. After cooling (to 50°), the reaction mixture was filtered (gravity) and the filtrate poured into 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).

Claim 31

Step 3: Preparation of 4'-Fluoro-3,3',5-trimethyl-1,1'-biphenyl-2-carboxaldehyde

Claim 32

Step 3: Preparation of 4'-Fluoro-3,3',5-trimethyl-1,1'-biphenyl-2-carboxaldehyde

Claim 33

In a 1 L, 3-neck flask equipped with a magnetic stirring bar, N 2 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 N 2 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.

Claim 34

In a 1 L, 3-neck flask equipped with a magnetic stirring bar, N 2 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 N 2 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.

Claim 35

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 N 21 triphenylphosphine (64.0 g, 0.24 mole) was added and the mixture was stirred for 30 minutes. The Grignard reagent was added by means of a dropping funnel in a slow, steady stream and the reaction mixture stirred for one hour. After addition of 200 ml 6N HCl, 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 of brine and dried over MgSO 4 . Filtration and evaporation yielded a black oil ( 70 g) which was chromatographed using a 120 mm "Still" column (1.5 kg of 230-400 mesh Silica Gel) and 40% (v:v) CH 2 C1 2 - hexane. There were collected 2 fractions of 1000 ml and 20 fractions of 500 ml, the pure product being found in fractions 5-14. Fractions 14-17 contained a mixture of the desired aldehyde plus 2,4-dimethylbenzaldehyde. There was obtained 19-21 g of combined product as a pale yellow solid ( 70% yield); one spot on tlc (R f =0.30 on Silica Gel GF plate with 40% (v:v) CH 2 C1 2 -hexane), mp 75-78°C. The mp of pure sublimed product is 78-80°.

Claim 36

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 N 21 triphenylphosphine (64.0 g, 0.24 mole) was added and the mixture was stirred for 30 minutes. The Grignard reagent was added by means of a dropping funnel in a slow, steady stream and the reaction mixture stirred for one hour. After addition of 200 ml 6N HCl, 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 of brine and dried over MgSO 4 . Filtration and evaporation yielded a black oil ( 70 g) which was chromatographed using a 120 mm "Still" column (1.5 kg of 230-400 mesh Silica Gel) and 40% (v:v) CH 2 C1 2 - hexane. There were collected 2 fractions of 1000 ml and 20 fractions of 500 ml, the pure product being found in fractions 5-14. Fractions 14-17 contained a mixture of the desired aldehyde plus 2,4-dimethylbenzaldehyde. There was obtained 19-21 g of combined product as a pale yellow solid ( 70% yield); one spot on tlc (R f =0.30 on Silica Gel GF plate with 40% (v:v) CH 2 C1 2 -hexane), mp 75-78°C. The mp of pure sublimed product is 78-80°.

Claim 37

Step 4: Preparation of 3-(4'-Fluoro-3,3',5-trimethyl-[1,1'-biphenyl]-2-yl)-2-propenal

Claim 38

Step 4: Preparation of 3-(4'-Fluoro-3,3',5-trimethyl-[1,1'-biphenyl]-2-yl)-2-propenal

Claim 39

A dry 1 L, 4-neck flask equipped with a magnetic stirring bar, N 2 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 ml) 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 N 2 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 aq. NaHC0 3 (150 ml) was added in a slow 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 MgSO 4 , filtered and evaporated.

Claim 40

A dry 1 L, 4-neck flask equipped with a magnetic stirring bar, N 2 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 ml) 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 N 2 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 aq. NaHC0 3 (150 ml) was added in a slow 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 MgSO 4 , filtered and evaporated.

Claim 41

The residual oil was taken up in THF (200 ml) and the solution treated with 6N HC1 (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 MgS0 4 . 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 CH 2 C1 2 -hexane (3:1, 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).

Claim 42

The residual oil was taken up in THF (200 ml) and the solution treated with 6N HC1 (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 MgS0 4 . 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 CH 2 C1 2 -hexane (3:1, 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).

Claim 43

Step 5: Preparation of Methyl (E)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-5-hydroxy-3-oxo-6-heptenoate

Claim 44

Step 5: Preparation of Methyl (E)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-5-hydroxy-3-oxo-6-heptenoate

Claim 45

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 dry THF (180 ml) at 0° (internal, keeping temperature 10°C) and under N 2 . 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[1,1'-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 10°C. The reaction mixture was stirred at 0°C for 30 minutes and then quenched by the slow addition of 6N HC1 (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 (MgS0 4 ), filtered and evaporated leaving a yellow oil ( 38 g); one major spot on tlc (Silica Gel GF) at Rf = 0.49 (5% (v:v) acetone-CH 2 C1 2 ).

Claim 46

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 dry THF (180 ml) at 0° (internal, keeping temperature 10°C) and under N 2 . 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[1,1'-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 10°C. The reaction mixture was stirred at 0°C for 30 minutes and then quenched by the slow addition of 6N HC1 (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 (MgS0 4 ), filtered and evaporated leaving a yellow oil ( 38 g); one major spot on tlc (Silica Gel GF) at Rf = 0.49 (5% (v:v) acetone-CH 2 C1 2 ).

Claim 47

Step 6: Preparation of Methyl (E)-7-(4'-Fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenoate

Claim 48

Step 6: Preparation of Methyl (E)-7-(4'-Fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenoate

Claim 49

Sodium tetrahydridoborate (1.92 g, 0.0507 mole) was added portionwise over a period of 10 minutes and with stirring to a cold solution (0°C) of methyl (E)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-5-hydroxy-3-oxo-6-heptenoate (37.5 g) in methanol (400 ml). The clear reaction mixture was stirred at 0°C for 15 minutes and then diluted with water (300 ml). While maintaining the temperature below 10°C with an ice-bath, the mixture was acidified with 12N HC1 ( 22 ml). Following extraction with ether (3 x 250 ml), the ether extracts were combined, washed with cold water and brine, dried (MgS0 4 ) and evaporated to a viscous oil ( 38 g). The product showed one major spot on tlc (silica gel G F ) with R f 0.21 (10% (v:v) acecone-CH 2 Cl 2 ).

Claim 50

Sodium tetrahydridoborate (1.92 g, 0.0507 mole) was added portionwise over a period of 10 minutes and with stirring to a cold solution (0°C) of methyl (E)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-5-hydroxy-3-oxo-6-heptenoate (37.5 g) in methanol (400 ml). The clear reaction mixture was stirred at 0°C for 15 minutes and then diluted with water (300 ml). While maintaining the temperature below 10°C with an ice-bath, the mixture was acidified with 12N HC1 ( 22 ml). Following extraction with ether (3 x 250 ml), the ether extracts were combined, washed with cold water and brine, dried (MgS0 4 ) and evaporated to a viscous oil ( 38 g). The product showed one major spot on tlc (silica gel G F ) with R f 0.21 (10% (v:v) acecone-CH 2 Cl 2 ).

Claim 51

Step 7: Preparation of (±) trans (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

Claim 52

Step 7: Preparation of (±) trans (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

Claim 53

A solution of methyl (E)-7-(3'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-h eptenoate ( 37.7 g, .0976 mole) and 1N sodium hydroxide (100 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. HC1 and extracted with ether (3 x 300 ml). The ether extracts were combined, washed with cold water and brine, dried over MgS0 4 , filtered, and evaporated to yield a yellow-orange oil ( 37 g).

Claim 54

A solution of methyl (E)-7-(3'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-h eptenoate ( 37.7 g, .0976 mole) and 1N sodium hydroxide (100 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. HC1 and extracted with ether (3 x 300 ml). The ether extracts were combined, washed with cold water and brine, dried over MgS0 4 , filtered, and evaporated to yield a yellow-orange oil ( 37 g).

Claim 55

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-CH 2 C1 2 . After collecting 3 fractions of 1 L and 25 fractions of 500 ml, elution was continued with 20% (v:v) acetone-CH 2 C1 2 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 Et 2 0-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 cis racemates.

Claim 56

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-CH 2 C1 2 . After collecting 3 fractions of 1 L and 25 fractions of 500 ml, elution was continued with 20% (v:v) acetone-CH 2 C1 2 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 Et 2 0-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 cis racemates.

Claim 57

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 with acetone-CH 2 C1 2 (1:9, v:v). Using the shave recycle technique, the trans isomer (5 g) and the cis racemate (4 g) were obtained. The samples of the trans racemate, collected from the two chromatographic separations, were combined to give a 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 standing.

Claim 58

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 with acetone-CH 2 C1 2 (1:9, v:v). Using the shave recycle technique, the trans isomer (5 g) and the cis racemate (4 g) were obtained. The samples of the trans racemate, collected from the two chromatographic separations, were combined to give a 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 standing.

Claim 59

Step 8: Preparation of (3R,5S)-N-((S)-a-Methylbenzyl)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenamide

Claim 60

Step 8: Preparation of (3R,5S)-N-((S)-a-Methylbenzyl)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenamide

Claim 61

A solution of (+)-trans-(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 (10 g, 28.2 mole) and (S)-(-)-a-methylbenzylamine (36.35 ml, 34.17 g, 282 mmole) in THF (35 ml) was stirred at gentle reflux under N 2 for 16 h. The reaction mixture was then cooled, diluted wit Et 2 0 (400 ml) and successively washed with H 2 0 (2 x 200 ml), 3N HC1 (2 x 300 ml) and brine (2 x 200 ml), dried (MgS0 4 ) and filtered. The filtrate was evaporated leaving the mixture of intermediate diastereomeric amides as a pale amber gum.

Claim 62

A solution of (+)-trans-(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 (10 g, 28.2 mole) and (S)-(-)-a-methylbenzylamine (36.35 ml, 34.17 g, 282 mmole) in THF (35 ml) was stirred at gentle reflux under N 2 for 16 h. The reaction mixture was then cooled, diluted wit Et 2 0 (400 ml) and successively washed with H 2 0 (2 x 200 ml), 3N HC1 (2 x 300 ml) and brine (2 x 200 ml), dried (MgS0 4 ) and filtered. The filtrate was evaporated leaving the mixture of intermediate diastereomeric amides as a pale amber gum.

Claim 63

The residual gum was digested in Et 2 0 (100 ml) and treated with hexane until turbid. This was seeded and cooled at 0°. Solid, 1.5 g, m.p. 110-12°, was collected by filtration. The filtrate was evaporated to a gum ( 12 g).

Claim 64

The residual gum was digested in Et 2 0 (100 ml) and treated with hexane until turbid. This was seeded and cooled at 0°. Solid, 1.5 g, m.p. 110-12°, was collected by filtration. The filtrate was evaporated to a gum ( 12 g).

Claim 65

This mixture of diastereomeric amides was chromatographed using a Waters Prep LC500. Separation of the mixture was accomplished using two prep PAK-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 obtained. Compound (2) which was 99% pure by HPLC is the desired diastereoisomer. Mother liquors from recrystallization of (2) were combined (1.9 g, 85% (2)).

Claim 66

This mixture of diastereomeric amides was chromatographed using a Waters Prep LC500. Separation of the mixture was accomplished using two prep PAK-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 obtained. Compound (2) which was 99% pure by HPLC is the desired diastereoisomer. Mother liquors from recrystallization of (2) were combined (1.9 g, 85% (2)).

Claim 67

Step 9: Preparation of (+)-(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

Claim 68

Step 9: Preparation of (+)-(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

Claim 69

A solution of N-((S)-a-methylbenzyl)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenamide (3.5 g, 7.35 mmol), diastereoisomer (2) in step 8 above, in EtOH (300 ml) containing 1N NaOH (44 ml, 44 mmole) and H 2 0 (44 ml) was refluxed under N 2 for 14 h. The solvent was removed in vacuo ( 40°). The residue was dissolved in ice water (300 ml) and Et 2 0 (500 ml), then cooled and stirred at 0° while 3N HC1 (50 ml) was slowly added. The organic layer was separated and washed successively with ice-cold 1N HC1 (200 ml) and brine (2 x 200 ml), dried (MgS0 4 ) and filtered. After evaporation of solvent in 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 chromatographed on a 50 mm "Still" column (230-400 mesh Silica Gel) using acetone-methylene chloride (10:90, 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 Et 2 0-hexane. The combined yield was 1.6 g, 61%. This was 99.99% pure by HPLC, m.p. 87-9°. + 40.56° ( C = 1.075, CHC1 3 ).

Claim 70

A solution of N-((S)-a-methylbenzyl)-7-(4'-fluoro-3,3',5-trimethyl[1,1'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenamide (3.5 g, 7.35 mmol), diastereoisomer (2) in step 8 above, in EtOH (300 ml) containing 1N NaOH (44 ml, 44 mmole) and H 2 0 (44 ml) was refluxed under N 2 for 14 h. The solvent was removed in vacuo ( 40°). The residue was dissolved in ice water (300 ml) and Et 2 0 (500 ml), then cooled and stirred at 0° while 3N HC1 (50 ml) was slowly added. The organic layer was separated and washed successively with ice-cold 1N HC1 (200 ml) and brine (2 x 200 ml), dried (MgS0 4 ) and filtered. After evaporation of solvent in 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 chromatographed on a 50 mm "Still" column (230-400 mesh Silica Gel) using acetone-methylene chloride (10:90, 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 Et 2 0-hexane. The combined yield was 1.6 g, 61%. This was 99.99% pure by HPLC, m.p. 87-9°. + 40.56° ( C = 1.075, CHC1 3 ).

Claim 71

A process for the preparation of (+)-(4R,6H)-(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 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 a-methyl benzylamides of the trans-racemate and separation of the diastereomers; and e) hydrolysis of the diastereomer containing the product (+)-trans enantiomer and lactonization.

Claim 72

A process for the preparation of (+)-(4R,6H)-(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 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 a-methyl benzylamides of the trans-racemate and separation of the diastereomers; and e) hydrolysis of the diastereomer containing the product (+)-trans enantiomer and lactonization.

Claim 73

The compound, (+)-(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, the corresponding dihydroxy acid, or a pharmaceutically acceptable salt thereof.

Claim 74

The compound, (+)-(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, the corresponding dihydroxy acid, or a pharmaceutically acceptable salt thereof.

Claim 75

A pharmaceutical composition comprising a pharmaceutical carrier and an effective amount of (+)-(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, the corresponding dihydroxy acid or a pharmaceutically acceptable salt thereof.

Claim 76

A pharmaceutical composition comprising a pharmaceutical carrier and an effective amount of (+)-(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, the corresponding dihydroxy acid or a pharmaceutically acceptable salt thereof.

Claim 77

A process for the preparation of (+)-(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 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 a-methyl benzylamides of the trans-racemate and separation of the diastereomers; and e) hydrolysis of the diastereomer containing the product (+)-trans enantiomer and lactonization.

Claim 78

A process for the preparation of (+)-(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 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 a-methyl benzylamides of the trans-racemate and separation of the diastereomers; and e) hydrolysis of the diastereomer containing the product (+)-trans enantiomer and lactonization.