NO174850B - 6- (4,4-bis (substituted or unsubstituted phenyl) -3- (1-methyl-1H-tetrazol-5-yl) -1,3-butadienyl) -2,2-disubstituted-1,3-dioxane 4-acetic acid or a metal salt thereof - Google Patents

Description

where R<*> and R<4> are each independently hydrogen, halogen, C 1-4 alkyl, <C> 1-4 alkoxy or trifluoromethyl, R<2>, R<3>, R<5> and R 6 are each independently hydrogen, halogen, C 1-4 alkyl or C 1-4 alkoxy, and R is hydrogen, a hydrolyzable ester group or a cation to produce a non-toxic pharmaceutically acceptable salt.

The terms "C 1-4 alkyl", "C 1-4 alkyl" and "C 1-4 alkoxy" herein denote (unless otherwise indicated) straight or branched alkyl or alkoxy groups, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, amyl, hexyl or the like. These groups preferably contain 1-4 carbon atoms, especially 1 or 2 carbon atoms. Unless otherwise indicated, the term "halogen" herein includes chlorine, fluorine, bromine, and iodine, while the term "halide" herein includes chloride, bromide, or iodide ions. The term "a cation for the preparation of a non-toxic pharmaceutically acceptable salt" herein includes non-toxic alkali metal salts, such as sodium, potassium, calcium or magnesium salts. The ammonium salt and salts with non-toxic amines, such as trialkylamines, dibenzylamine, pyridine, N-methylmorpholine, N-methylpiperidine and other amines which are used for the preparation of salts of carboxylic acids. Unless otherwise indicated, the term "a hydrolyzable ester group" as used herein includes an ester group which is physiologically acceptable and hydrolyzable under physiological conditions, such as C 1-6 alkyl, phenylmethyl or pivaloyloxymethyl.

In the compounds of formula (I), (II), (XI) and (XII) it is intended that all double bonds should have a trans configuration, i.e. (E), as shown in the structural formulas used here. In the compounds of formula (IV), (V), (VI), (VII), (VIII) and (IX), on the other hand, the intention is that the configuration of the double bonds is trans, cis or mixtures thereof, i.e. (E), (Z), when n is 0 and (E)(E), (Z)(Z), (E)(Z) and (Z)(E), when n is 1.

Due to the fact that the compounds according to the present invention have two asymmetric carbon atoms, the present invention includes the enantiomeric and diastereomeric forms of the intermediates used in the methods for preparing compounds of formula (I) and (II) which are described herein. The compounds of formula (I) and (II), which contain 2 asymmetric centers, can produce 4 possible stereoisomers named RR, RS, SR and SS enantiomers. The compounds of formula (I) which contain two asymmetric carbon atoms, which carry the hydroxy group in the 3- and 5-position, can in particular have 4 possible stereoisomers, which are named (3R, 5S), (3S,5R), (3R,5R) and (3S,5S) stereoisomers. As used herein, the term "erythro" includes a mixture of (3R,5S) and (3S,5R) enantiomers, and the term "threo" includes a mixture of (3R,5R) and (3S,5S) enantiomers. The use of a single designation, such as (3R.5S) most often denotes one stereoisomer. The lactone compounds of formula (II) also contain 2 asymmetric carbon atoms in the 4- and 6-position, and the resulting 4 stereoisomers can be named (4R,6S), (4S,6R), (4R,6S) and (4S,6S) stereoisomers . As used herein, the term "trans"-lactone encompasses a mixture of (4R,6S) and (4S,6R) enantiomers, while the term "cis"-lactone encompasses a mixture of (4R,6R) and (4S,6S) enantiomers. The use of a simple designation such as (4R,6S) most often denotes one enantiomeric lactone compound.

The substituted 1,3-dioxane compounds of formula (Illa), (Illb) and other similar compounds described herein also contain 2 asymmetric carbon atoms in the 4- and 6-position as shown below

and the resulting 4 stereoisomers may be termed (4R,6S), (4S,6R), (4R,6R) and (4S,6S) stereoisomers. As used herein, the term "trans"-1,3-dioxane includes a mixture of (4R,6R) and (4S,6S) enantiomers, while the term "cis"-1,3-dioxane includes a mixture of (4R,6S) and (4S,6R) enantiomers. Since the most preferred enantiomer of the lactone compounds of formula (II) happens to have the same (4R,6S) stereoisomeric designation as the most preferred enantiomer of the 1,3-dioxane intermediates of the present invention, the additional designation of "trans" or "cis" is included in the present to avoid possible entanglement.

In the compounds of formula (Illa) and (Illb) R<9> and R<10> are each C1-4alkyl, or R<9> and R<10> are, together with the carbon atom to which they are attached, cyclopentyl, cyclohexyl or cycloheptyl. R<9> and R<10> are each preferably methyl, or R<9> and R<1>^ together with the carbonate to which they are attached, are cyclohexyl. R<12> is preferably hydrogen, methyl or a metal cation, especially lithium. The cis isomer of the compounds of formula (Illa) is preferred, and the cis-(4R, 6S) isomer of the compounds of formula (Illb) is the most preferred.

The antihypercholesterolemic compounds of formula (I) and (II) can be prepared by various methods and preferably by using the intermediates of formula

essentially in the cis form,

where R<9>, R10 and R<12> have the meaning given above.

The compounds of formula (Illa) and (Illb) can be prepared by reacting an aldehyde of formula (IV) with an ester of acetoacetic acid and then reacting a ketone or ketal with a compound of formula (VI) followed by hydrolysis of the resulting 1, 3-dioxane of formula (VII) and optionally dissolve the acid of formula (VIII), as shown in reaction scheme 1. In reaction core 1, e'7 and R<8> are each independently hydrogen, C-L-^alkyl or phenyl, which optionally is substituted by one or two of the substituents C 1-4 alkyl, halogen, C 1-4 alkoxy or trifluoromethyl, R 11 is a hydrolysable ester group, n is 0 or 1, and R<9> and R 10 have the meaning given above. The keto ester of formula (V) can be prepared by reacting an ester of acetoacetic acid with an aldehyde of formula (IV) by a method known in the art in an inert organic solvent, such as tetrahydrofuran, at temperatures from approx. 0°C to approx. -78" C in the presence of a base, such as sodium hydride, lithium diisopropylamide or n-butyllithium.

The starting materials of formula (IV), where n is 0 or 1, are known, or can be easily prepared by known methods. The starting materials of formula (IV), where n is 1, can also be prepared by reacting the compounds of formula (IV), where n is 0, with Wittig reagents, such as triphenylphosphoranylidene acetaldehyde, and by other methods known in the art. It is obvious to the person skilled in the art that the relative configuration of the double bond (n=0) or double bonds (n=1) in the starting materials of formula (IV) can be trans, cis or mixtures thereof. The relative amounts of each geometric isomer (E) or (Z) are determined by the commercial availability or by the reaction conditions used in the preparation. In a specific example described herein, a mixture containing mostly the trans (E) isomer is used. Although a small percentage of the other isomer may be present in the reactions shown in reaction core 1, it is obvious to the person skilled in the art that the relative amount of isomers is not critical, as the double bond is oxidized and thereby removed in the ozonolysis reaction.

The keto ester of formula (V) can be reduced to the dihydroxy ester of formula (VI) by reduction of the ketone group with reducing agents known in the art. The reduction is preferably carried out in a stereospecific manner by a two-stage stereospecific reduction in order to maximize the production of the preferred erythroisomer of the dihydroxyester of formula (VI). The stereospecific reduction is carried out with trisubstituted alkylboranes, preferably triethylborane or tri-n-butylborane or alkoxydialkylboranes, preferably methoxydiethylborane or ethoxydiethylborane [Tetrahedron Letters, 28, 155 (1987)] at a temperature from approx. -70°C to ambient temperature. The complex that is produced is then reduced with sodium borohydride at a temperature from approx. -50°C to approx. -78°C in an inert organic solvent, such as tetrahydrofuran, diethyl ether or 1,2-dimethoxyethane, preferably tetrahydrofuran. The reduction is then terminated by the addition of methanol with or without the addition of aqueous hydrogen peroxide and a buffer agent. Some of the compounds of formula (VI) are known, and are described in US Patent Nos. 4,248,889 and 4,650,890.

The compounds of formula (VII) can be prepared from the compounds of formula (VI) by reacting a ketone, such as 2-propanone, 3-pentanone, cyclopentanone or cyclohexanone in a suitable inert organic solvent, e.g. toluene, benzene or xylene at temperatures from approx. 20° C to the cooling temperature of the solvent used, in the presence of a small amount of organic acid, mineral acid or resin acid, e.g. p-toluenesulfonic acid or sulfuric acid and optionally remove the water that is formed with a desiccant, e.g., Na2SC>4, MgSO4 and molecular sieves or by azeotropic removal with a Dean-Stark trap or a similar device. The reaction of a compound of formula (VI) with a ketone can also be carried out without a solvent. Alternatively, the reaction of compounds of formula (VII) described above can be carried out with a ketal, such as 2,2-dimethoxypropane, 1,1-dimethoxycyclohexane and the like.

The compounds of formula (IIa) where R^<2> is a hydrolyzable ester group, and preferably C-L-4 alkyl, can be prepared from the corresponding compounds of formula (VII) by oxidation of the olefin group to an aldehyde group in a conventional manner. Alternatively, a compound of formula (VII) is first hydrolyzed by basic hydrolysis to a compound of formula (VIII), which is then oxidized to obtain a compound of formula (Illa), where R<*2> is hydrogen. A particularly suitable oxidation method is reaction of a compound of formula (VII) or (VIII) in an inert organic solvent, such as methanol, ethyl acetate or methylene chloride with ozone at temperatures of approx. -50°C to approx. -78°C. When the reaction with ozone has come to an end, which can be seen from the color of the reaction mixture, the ozonide intermediate product is broken down by adding a mild reducing agent, e.g. dimethylsulfide or triphenylphosphine, to obtain the desired aldehyde of formula (Illa).

The preferred cis-(4R,6S) aldehydes of formula (IIIb) can be prepared from the corresponding racemic acid of formula (VIII) by conventional cleavage methods, such as fractional crystallization, after addition of a suitable salt-forming group. The resulting mixture of diastereoisomeric salts formed with an optically active salt-forming agent, such as (IS,2R)-ephedrine or α-methylbenzylamine, is separated and the separated, dissolved salt is converted to a compound of formula (Illb).

The salt-forming agent is preferably (IS,2R)-ephedrine and the separation method is fractional crystallization. The cleavage can be carried out in an inert organic solvent, and preferably in a mixture of hydrocarbon-alcohol solvent, e.g. hexanemethanol mixture, where the dissolved salt can be crystallized from the solution. If desired, the acid of formula (IIIb) can be converted into a salt, where R<12> is a metal cation, or into a hydrolyzable ester group where R12 is C 12 alkyl.

The antihypercholesterolemic compounds of formula (I) and (II) can be prepared from a compound of formula (Illa) or (Illb) by those in US patent application no. 018,542, DK patent application no. 972/88, TJS patent application no. 018,558 and DK patent application no. 973/88 described methods. The use of aldehydes with formula (Illa) is shown in reaction core 2 and the use of chiral aldehydes with formula (Illb) is shown in reaction scheme 3. In reaction schemes 2 and 3, R<1>, R<2>, R23, R<4 >, R5, R<6>, R9, r10 and R<i2> the meaning given above, and Z is where R13 is <C>1_4alkyl, R*<4> is phenyl, which is unsubstituted or substituted by one or two C 1-4 alkyl or chlorine substituents, and X is bromine, chlorine or iodine. The phosphonium salt of formula (X) and the phosphonate of formula (X) are described herein in US Patent Application No. 018,558 and DK Patent Application No. 973/88. The reaction of a compound of formula (X) with a compound of formula (Illa) or formula (Illb) to produce a compound of formula (XI) or (XII), respectively, where R<12> is C1-4alkyl, can be carried out in a inert organic solvent, such as tetrahydrofuran or N,N-dimethylformamide, in the presence of a strong base, such as n-butyllithium at a temperature from about -50°C to approx. -78"C. When the reaction of a compound of formula (X) is carried out with a compound of formula (Illa) or (Illb), where R<12> is hydrogen, it is preferred to use two equivalents of a strong base, so as n-butyl lithium. Alternatively, the salt of a compound of formula (Illa) or (Illb) can be prepared, which is then treated with a compound of formula (X) and a strong base. The addition methods, salt formation methods and ylide preparation are known to those skilled in the art. The tetrazole compounds of formula (XI) or (XII) can be easily deprotected by known methods, such as a mild acid, for example 0.2 N HCl and 0.5 N HCl in an inert organic solvent, such as tetrahydrofuran, for the production of erythro compounds of formula (Ia) or the (3R, 5S) compounds of formula (Ib), which can then be converted into the trans compounds of formula (II) or the (4R, 6S) compounds of formula (II) on a method known to the person skilled in the art.

In vivo acute cholesterol biosynthesis inhibition in rats.

Male Wistar rats (160-200 g, 2 rats per cage) are maintained on a normal diet (Purina rat chow and water ad libitum) for at least 7 days in a reverse lighting condition (dark from 7 am to 5 pm). The lining was removed 15 hours before dosing. The connections were administered at 8 in the morning by intragastric intubation using 0.5-1.0 ml of water or propylene glycol solutions of the sodium salt, lactones or esters of the test compounds. The control groups received the same volume of vehicle. 30 minutes after administration of the test compounds, the rats were injected intraperitoneally with 0.9 ml of 0.9$ NaCl containing approx. 120 jjCi per kg body weight sodium [1-<14>C]acetate (1-3 mCi/mmol). After an incubation period of 60 minutes, the rats were euthanized and liver and blood samples were taken. Aliquots of plasma (1.0 ml) obtained by centrifugation of heparin + EDTA-treated blood and aliquots of liver homogenates (equivalent to 0.50 g liver wet weight) were taken for determination of radiolabelled 3-hydroxysterols. Sterol isolation of the liver samples was according to the method of Kates described in Techniques in Lipidology (M. Kates, ed.) pp. 349, 360, 363, North Holland Publ. Co. Amsterdam, 1972, while the plasma samples were directly saponified followed by isolation of digitonin-precipitable sterols. The amount of <14>C-labeled sterols was determined by liquid scintillation counting (corrected for efficiency). The average percentage inhibition of <14>C incorporated into liver and plasma cholesterol was calculated for groups of treated animals and compared with average values for control experiments which were carried out at the same time.

The above test therefore provides information on the ability of the test compounds to express the de novo biosynthesis of cholesterol in vivo in rats by oral dosing. During the execution of the above test, e.g. the compound of Example 13 a 50$ inhibitory dose (ED50) for both plasma and liver cholesterol comparable to values obtained for mevinolin (lovastatin) using a similar method [Alberts et al., Proe. nati. Acad. Sei, 77, 3957-3961

(1980)].

In the following examples, all temperature indications are in degrees Celsius. Melting points are recorded on a Thomas-Hoover capillary melting point apparatus and are uncorrected. Proton magnetic resonance spectra (<*>H NMR) were recorded on a Bruker "Am 300", Bruker "WM 360" or Varian "T-60 CW" spectrometer. All spectra are determined in CDCl3, DMSDO-d^ or D2O, unless otherwise indicated and chemical shifts are given in S units derived from the internal standard tetramethylsilane (TMS) and interproton coupling constants are given in Hertz (Hz). The splitting patterns are denoted as follows: s, singlet; d, doublet, t, triplet; q, quartet; m, multiplet; br, broad peak; dd, double duplicate; and dq, double quartet. Carbon-13 nuclear magnetic resonance spectra (<13>C NMR) were recorded on a Bruker "AM 300" or Bruker "WM 360" spectrometer and were broadband proton coupled. All spectra were determined in CDCl 3 , DMSO-dfc or DgO, unless otherwise indicated by internal deuterium lock, and chemical shifts were indicated in S units precipitated from tetramethylsilane. Infrared (IR) spectra were recorded on a Nicolet "MX-1 FT" spectrometer from 4000 cm"<1 >to 400 cm--'-, calibrated to 1601 cm-<*> absorption using a polystyrene film and were indicated in reciprocal centimeters (cm-<1>). Relative intensities are indicated as follows: s (strong), m (medium), and w (weak). Optical rotations [a]§<5> were determined on a Perkin-Elmer 241 polarimeter in CHCl3 at the indicated concentrations.

Analytical thin layer chromatography (TLC) was performed on pre-coated silica gel plates (60F=254) and visualized using UV light, iodine vapor and/or staining with one of the following reagents: (a) methanolic or ethaniolic phosphomolybdic acid (2%) and heating, (b) reagent (a) followed by 2% cobalt sul f at in 5 M H2SO4 and heating. Column chromatography, also called flame column chromatography, is carried out on a glass column using finely divided silica gel (32-63 pm on silica gel-H) and pressure which was slightly above atmospheric pressure with the stated solvents. Ozonolysis reactions are carried out using a Welsbach ozonator type "T-23". All evaporation of solvents is carried out under reduced pressure. As used herein, the term "hexanes" means a mixture of isomeric C 1 -hydrocarbons as specified by the American Chemical Society, and the term "inert" atmosphere means an argon or nitrogen atmosphere unless otherwise indicated.

EXAMPLE 1

Cis- 2, 2- dimethyl- 6-( 2- f envletenvl 1 - 1 . 3- dioxane- 4- acetic acid methyl ester

Methyl 3,5-dihydroxy-7-phenyl-6-enoate (98% diastereomeric purity) (2.37 g, 9.48 mmol) was stirred with 2,2-dimethoxypropane (20 mL) and a catalytic amount of p -toluenesulfonic acid for 16 hours. The solution was partitioned between diethyl ether and a dilute aqueous sodium bicarbonate solution. The organic phase was dried (Na 2 SO 4 ) and concentrated under reduced pressure to give a yellow solid. After recrystallization from isopropyl ether, 1.70 g (62$) of the title compound was obtained as a white solid, m.p. 84-86.5°C.

Alternatively, 0.2 g of solid sodium carbonate can be added to the 2,2-dimethoxypropane solution and the solution stirred vigorously. The solid is filtered through a fluted filter paper. Excess 2,2-dimethoxypropane is removed under reduced pressure to give a yellow solid, which is recrystallized from isopropyl ether.

%-NMR (CDCl 3 ) S : 7.37-7.19 (5H, m), 6.59 (1H, d, J = 15.9 Hz), 6.14 (1H, dd, J = 15.9 , 6.4 Hz), 4.57-4.35 (1H, m), 4.42- 4.35 (1H, m), 3.68 (3H, s), 2.58 (1H, d, J = 15.6, 6.9 Hz), 2.14 (1H, dd, J =) 15.6, 6.3 Hz), 1.74-1.61 (1H, m), 1.52 (3H, s), 1.43 (3H, s), 1.45-1.35 (1H, m).

Analysis for Ci7<H>22°4<:>

Calculated: C: 70.32$, H: 7.63$.

Found: C: 70.24$, H: 7.69$.

EXAMPLE 2

Cis- 2, 2- dimethyl- 6-( 2- phenylethenyl)- 1. 3- dioxane- 4- acetic acid

A solution of 2,2-dimethyl-6-(2-phenylethenyl)-1,3-dioxane-4-acetic acid methyl ester (8.5 g, 29.3 mmol) in 1N NaOH (32 mL) and methanol (64 mL ) was heated to reflux for 45 min. After evaporation under reduced pressure, the aqueous solution was washed once with diethyl ether and acidified with 1N HCl (33 mL). The precipitate was collected and recrystallized from ethyl acetate/isopropyl ether to give 7.2 g (90%) of the title compound as a colorless solid, m.p. 153-155°C. -^H-NMR (CDCl 3 ) S: 7.37-7.20 (5H, m), 6.60 (1H, d, J = 16.0 Hz), 6.14 (1H, dd, J = 16 ,0, 6.4 Hz), 4.59-4.54 (1H, m), 4.43- 4.35 (1H, m), 2.62 (1H, dd, J = 16.0, 7.2 Hz), 2.51 (1H, dd, J = 16.0, 5.3 Hz), 1.77-1.72 (1H, m), 1.54 (3H, s), 1.46 (3H , s), 1.50-1.36 (1H, m).

Analysis for C^^<H>2o°4<:>

Calculated: C: 69.54$, H: 7.30$

Found: C: 69.20$, H: 7.33$.

EXAMPLE 3

Cleavage of cis- 2. 2- dimethyl- 6-( 2- phenylethenyl)- 1. 3- dioxane-4- acetic acid

The racemic cis-2,2-dimethyl-6-(2-phenylethenyl)-1,3-dioxane-4-acetic acid (0.31 g, 1.1 mmol) (prepared in Example 2), was dissolved in a boiling hexane/ethanol solution containing (1,2R)-ephedrine (0.2 g, 1.1 mmol). The resulting solution was very slowly brought to room temperature to obtain 0.21 g (41.4%) of a colorless chiral salt (use of diastereomerically pure seed crystal is recommended during the resolution) m.p. 170-171"C.

The chiral acid was liberated by acidic workup (as described in Example 4) and its enantiomeric purity was determined to be 100% by 1 H-NMR using L-phenyltrifluoromethylcarbinol as a chiral solvent.

[a]<g5> = +5.45' (c = CHC13).

EXAMPLE 4

Cis-(4R,6S)-2,2-dimethyl-6-formyl-1,3-dioxane-4-acetic acid

The cleaved salt of cis-2,2-dimethyl-6-(2-phenylethenyl)-1,3-dioxane-4-acetic acid and (1S,2R)-ephedrine (6.6 g, 14.9 mmol)

(prepared in Example 3) was partitioned between 0.5 N HCl (30 mL) and diethyl ether. The ether phase was washed with brine, dried (MgSO 4 /Na 2 SO 4 ) and concentrated under reduced pressure to give 4.1 g (99.6%) of the free acid. This acid was dissolved in dry methylene chloride (100 mL) and ozone was passed through the solution at -78°C, until deep blue coloration. Excess ozone was removed by purging with nitrogen and the ozonide formed was decomposed by adding CH 3 SCH 3 (5 ml) and heating the solution to room temperature and standing for 16 hours. The solution was concentrated under

reduced pressure, and the residue was dissolved in isoamyl ether (ca< 100 ml). Benzaldehyde formed during the ozonolysis was removed azeotropically with isoamyl ether under reduced pressure to afford the title compound.

<1>H-NMR (CDCl3) S: 9.57 (1H, s), 4.40-4.30 (2H, m), 2.60 (1H, dd, J = 16.0, 7.0 Hz), 2.49 (1H, dd, J = 16.0, 6.0 Hz), 1.88-1.83 (1H, m) 1.49 (3H, s), 1.46 (3H, s), 1.42-1.31 (1H, m).

EXAMPLE 5

Dimetvl- r3. 3-bis(4-fluorophenyl)-2-(1-methyl-1H-tetrazole-5-yl)-2-propene-1-ylphosphonate

A slurry of 3,3-bis-(4-fluorophenyl)-1-bromo-2-(1-methyl-1H-tetrazol-5-yl)-2-propane (1.17 g, 3.0 mmol) and Trimethylphosphite (0.41 g, 3.3 mmol) was heated at 100°C for 5 min. After cooling to room temperature, excess trimethylphosphite was removed in vacuo to provide a pale yellow solid. This solid was recrystallized from an ethyl acetate/hexane mixture to provide the title compound as a pure white solid, m.p. 140-141°C.

IR (KBr) <N/>max: 1604, 1511 cm -1 .

<1>H-NMR (CDCl 3 ) δ: 7.7-6.8 (8H, m), 3.6 (3H, s), 3.5 (3H, s), 3.42 (3H, s) , 3.2 (2H, d).

Analysis for C1gH1gF203N4<P:>

Calculated: C: 54.29$, H: 4.56$, N: 13.33$.

Found: C: 53.83$, H: 4.48$, N: 13.50$.

EXAMPLE 6

Cis-( 4R . 6S1- 6- T4 . 4- bis( 4- fluorophenyl)- 3-( 1- methyl- 1H- tetrazol-5-yl 1- 1. 3- butadienyl- 2, 2- dimethyl- 1. 3- dioxane- 4- acetic acid

The crude chiral acid prepared in Example 4 was dissolved in dry THF (50 mL) and the resulting solution was transferred to a 250 mL three-necked flask purged with nitrogen and fitted with a mechanical stirrer. After vigorous stirring of the solution and cooling to -78°C, n-BuLi (2.5 M in hexane, 5.96 mL) was added dropwise. At the end of the addition, the solution became a suspension of a white solid-like gel.

A separate flask containing dimethyl-[3,3-bis-(4-fluoro-phenyl)-2-(1-methyl-1H-tetrazol-5-yl)-2-propen-l—yl]phosphonate (6, 2 g, 14.7 mmol) (prepared in Example 5) in THF (50 mL) was cooled under a nitrogen atmosphere to -78°C and n-BuLi (2.5 M in hexane, 5.96 mL) was slowly added. The resulting red-brown solution was stirred for 15 minutes at -78°C. The phosphonate anion solution was transferred through a double-ended needle to the above vigorously stirred suspension at -78°C containing the lithium salt of the chiral acid. After addition, the resulting brown solution was stirred for 30 minutes at -78°C and 16 hours at room temperature. The THF solution was partitioned between 0.5 N HCl and ethyl acetate. The organic phase was washed with brine (2x), dried (Na 2 SO 4 ) and concentrated under reduced pressure. The residue was chromatographed on silica gel (66:33:1/diethyletherhexane:acetic acid) to provide 3.80 g (51.6% total yield from the original ephedrine salt, toluene was used for azeotropic removal of the residual acetic acid) of the title compound as a yellow foam. [a]<B5> = +106.1° c=2.23, CHC13).

<1->H-NMR (CDCl3) §: 7.24-6.82 (8H, m), 6.62 (1H, d, J = 15.0 Hz), 5.32 (1H, dd, J = 15.0, 5.7 Hz), 4.42-4.37 (1H, m) 4.30-4.23 (1H, m), 3.51 (3H, s), 2.53 (1H , dd, J = 15.9, 7.0

Hz), 2.42 (1H, dd, J = 15.9, 5.6 Hz), 1.62-157 (1H, m), 1.46 (3H, s), 1.33, (3H, s), 1.30-1.20 (1H, m).

EXAMPLE 7

Trans-( 4R. 6S)- 6- r4. 4-bis(4-fluorophenyl)-3-(1-methyl-1H-tetrazol-5-vl)-1.3-butadienyl-tetrahydro-4-hydroxy-2H-pyran-2-one

Cis-(4R,6S)-6-[4,4-bis(4-fluorophenyl)-3-(1-methyl-1H-tetrazol-5-yl)-1,3-butadienyl]-2, 2-Dimethyl-1,3-dioxane-4-acetic acid (3.7 g, 7.45 mmol) was dissolved in a solution of THF (90 mL) and 0.2N HCl (60 mL) and allowed to stand in 16 hours. The solution was partitioned between ethyl acetate and water. The organic phase was washed with brine (2x), dried (Na 2 SO 4 ) and concentrated under reduced pressure. The residue was dissolved in dry methylene chloride (60 mL) and stirred for 4 h in the presence of 1-cyclohexyl-3-(2-morpholinomethyl)carbodiimide-metho-p-toluenesulfonate (6.6 g, 15.6 mmol). The solution was concentrated under reduced pressure, and the residue partitioned between ethyl acetate and water. The organic phase was dried (Na 2 SO 4 ) and concentrated under reduced pressure. The residue was purified by chromatography on silica gel (1:1/ethyl acetate diethyl ether). After recrystallization from ethyl acetate-hexane, 1.33 g (40.1%) of the title compound was obtained as a white solid, m.p. 172-173°C.

[ot]2.5 = +237.8° (c-2.17, CHCl 3 ).

EXAMPLE 8

Methyl- 3- hydroxy- 5- oxo- 6. 8- decadienoate

To a cold (-30°C) solution of methyl acetoacetate (41.5 g, 357 mmol) in THF (500 mL) was added lithium diisopropylamide (476 mL, 1.5 M solution in cyclohexane, 714 mmol). The resulting solution was stirred for 15 minutes at -30°C. After cooling to -78°C, 2,4-hexadienal (34.3 g, 357 mmol) was added and the solution was stirred for 10 minutes at -78°C and 16 hours at room temperature. The solution was concentrated under reduced pressure and the remaining syrup was partitioned between 1N HCl and ethyl acetate. The organic phase was washed with brine (2x), dried (Na 2 SO 4 ) and concentrated. The residue was purified by chromatography on silica gel (diethyl ether:hexane/2:1) to provide 18.5 g (24.4%) of the title compound as an oil.

<1>E-NMR for (E) (E) isomer (200 MHz, CDCl 3 ) S; 6.3 (1H, dd, J = 14.7, 11.9 Hz), 6.02 (1H, dd, J = 14.7, 11.9 Hz), 5.75 (1H, dq, J = 14.7, 6.4 Hz), 5.5, (1H, dd, J = 18.7, 6.4 Hz), 4.74-4.5 (1H, m), 3.73 (3H, s), 3.51 (2H, s), 2.6 (2H, d, J = 5.8 Hz), 1.77 (3H, d, J = 6.4 Hz).

EXAMPLE 9

Methyl- 3. 5- dihydroxy- 6, 8- decadienoate

To a cold (-15°C) solution of methyl 3-hydroxy-5-oxo-6,8-decadienoate (18.5 g, 86.9 mmol) in THF (300 mL) was added triethylborane (1 M in THF , 113 mL, 113 mmol) was added, and the solution was stirred for 20 minutes. After cooling the mixture to -78°C, NaBH 4 (6 g, 159 mmol) and methanol (37.5 mL) were added. The solution was vigorously stirred for 30 minutes at -78°C and at room temperature for 3 hours. The solvent was removed under reduced pressure and the residue was partitioned between 1N HCl and ethyl acetate. The organic phase was dried (Na 2 SO 4 ) and concentrated. The residue was purified by silica gel chromatography (diethyl ether hexane, 3:1) to provide 7.95 g (42.7 g) of the title compound as a yellow oil.

<1->H-NMR for (E) (E) isomer (360 MEz, CDCl 3 ) S: 6.18 (1H, dd, J = 15.1, 10.4 Hz), 6.00 (1H, dd , J = 15.1, 10.4 Hz), 5.69 (1H, dq, J = 15.1, 7.0 Hz), 5.52 (1H, dd, J = 15.1, 6.7 Hz), 4.46-4.37 (1H, m), 4.29-4.22 (1H, m), 3.69 (3H, s) 2.60-2.42 (2H, m), 1.72 (3H, d, J = 7.0 Hz), 1.74-1.57 (2H, m).

EXAMPLE 10

Methyl cis-4-(1,3-pentadienyl)-1,5-dioxaspirof 5. 5~ lundecan-2-acetate

Methyl 3,5-dihydroxy-6,8-decadienoate (7.6 g, 35.5 mmol) and p-toluenesulfonic acid (0.1 g) were added to cyclohexanone (10 g, 100 mmol) and stirred for 16 h at room temperature. The yellow solution was directly applied to a silica gel column and the product was eluted with diethyl ether hexane (1:4). The appropriate fractions were combined to provide 3.52 g (33.6$) of the title compound as a colorless oil.

<1>H-NMR for (E) (E) isomer (360 MHz, CDCl 3 ) S: 6.16 (1H, dd, J = 15.1, 10.6 Hz), 6.00 (1H, dd, J = 15.1, 10.6 Hz), 5.71-5.65 (1H, dd, J = 15.1, 6.5 Hz), 5.47 (1H, dd, J = 15.1, 6.4 Hz), 4.44-4.39 (1H, m), 4.35-4.30 (1H, m), 3.66 (3H, s), 2.52 (1H, dd, J = 1.54, 7.9 Hz), 2.30 (1H, dd, J = 15.4, 6.5 Hz), 2.1-1.18 (12 H, m), 1.72 (3H , d, J = 6.5 Hz).

Analysis for C-L7<H>26<O>4:

Calculated: C: 69.36$, H: 8.90$

Found: C: 69.59$, H: 9.16$.

EXAMPLE 11

Cis-4-(1,3-pentadienyl)-1,5-dioxaspiror5,5-lundecane-2-acetic acid

Methyl 4-(1,3-pentadienyl)-1,5-dioxaspiro[5,5]undecane-2-acetate (3.5 g, 12.4 mmol) was heated to reflux in a solution of 1 N NaOH (13 ml) and methanol (26 ml). Methanol was removed under reduced pressure and the remaining aqueous solution was acidified with 1N HCl and extracted with diethyl ether. The organic phase was dried (Na 2 SO 4 ) and concentrated. The remaining solid was recrystallized from ethyl acetate/hexane to provide 2.0 g (55.9 g) of the title compound as a colorless solid, m.p. 144-146.5°C.

<1->H-NMR (360 MHz, CDCl 3 ) S: 6.18 (1H, dd, J = 18.0, 12.5 Hz), 5.72 (1H, dq, J = 18.0, 7 .7 Hz), 5.99 (1H, dd, J = 18.0, 12.5 Hz), 5.48 (1H, dd, J = 18.0, 7.6 Hz), 4.45-4 .37 (1H, m), 4.37-4.25 (1H, m), 2.56 (1H, dd, J = 18.9, 8.8 Hz), 2.48 (1H, dd, J = 18.9, 6.1 Hz), 2.60-1.30 (12 H, m), 1.73 (3H, d, J = 7.7 Hz).

Analysis for C15<H>24<O>4:

Calculated: C: 68.54$, H: 8.62$.

Found: C: 68.36$, H: 8.55$.

EXAMPLE 12

Cis-4-r4. 4-bis(4-fluorophenyl)-3-(1-methyl-1H-tetrazol-5-yl)-1. 3- butadienyl - 1. 5- dioxaspiror5 . 5~ lundecan- 2- acetic acid A. 4- formyl- l, 5- dioxaspiror5. 5 lundecan- 2- acetic acid

Ozone was passed through a solution of 4-(1,3-pentadienyl)-1,5-dioxaspiro[5,5]undecane-2-acetic acid (570 mg, 2.0 mmol) in methylene chloride (25 mL) at -78 °C. After the solution had turned blue, nitrogen was passed through the solution to remove excess ozone. Dimethyl sulfide (0.5 mL) was added and the solution concentrated under reduced pressure to provide the title compound as a viscous oil, which was used without further purification in the subsequent step.

<1>H-NMR (60 MEz, CDCl 3 ) S : 9.57 (1H, s), 4.52-4.14 (2E, m), 2.60-2.31 (2H, m), 2 .10-1.10 (12 H, m).

B. Cis-4-r4. 4-bis(4-fluorophenyl)-3-(1-methyl-1E-tetrazol-5-yl)-1.3-butadienyl-1.5-dioxaspiro-r5. 5~ lundecan- 2- acid

To a solution of dimethyl [3,3-bis(4-fluorophenyl)-2-(1-methyl-1E-tetrazol-5-yl)-2-propenyl]phosphonate (1.7 g, 4 mmol) in TEF ( 20 mL) at -78°C, n-Buli (1.6 mL, 4 mmol, 2.5 M in hexane) was added. The resulting brownish-red solution was stirred for 30 minutes at -78°C. Using a double-ended needle, the solution was transferred to a solution containing 4-formyl-1,5-dioxaspiro[5,5]undecane-2-acetic acid (prepared in step A) in TEF (10 mL) and kept at -78° C. After completion of the transfer, the overall reaction mixture was stirred at -78° C. for 1 hour, and at room temperature for 4 hours. The solution was then partitioned between 0.5 N EC1 and ethyl acetate. The organic phase was washed with brine (2x), dried (Na 2 SO 4 ) and concentrated under reduced pressure. The residue was purified by chromatography on silica gel (diethyl ether:-hexane:acetic acid/50:20:1) to provide 342 mg (31.9% overall yield) of the title compound as a yellow foam. I<->E.NMR (360 MHz, CDCl3) S: 7.25-6.84 (8E, m), 6.66 (1E, d, J = 16.0 Ez), 5.32 (1E, dd, J = 16.0, 5.10 Ez) , 4.45-4.25 (2E, m), 3.52 (3E, s), 2.56 (1E, dd, J = 16.0, 7.6 Ez), 2.44 (1E, dd, J = 16.0, 5.1 Ez), 1.89-1.17 (12E, m).

EXAMPLE 13

Trans-6-T4,4-bis(4-fluorophenyl)-3-(1-methyl-1H-tetrazol-5-yl)-1. 3- butadienyl- tetrahydro- 4- hydroxy- 2H- pyran- 2- one

A mixture of 4-[4,4-bis(4-fluorophenyl)-3-(1-methyl-1H-tetrazol-5-yl)-1,3-butadienyl]-1,5-dioxaspiro[5,5] Undecane-2-acetic acid (280 mg, 0.52 mmol) in 20 mL TEF/0.5 N HCl (1:1) was left at ambient temperature for 26 h. The solution was partitioned between saline and ethyl acetate. The organic phase was washed with brine (2x), dried (Na 2 SO 4 ) and concentrated. The resulting foam (126 mg) was dissolved in dry methylene chloride (10 mL) and treated with 1-cyclohexyl-3-(2-morpholinomethyl)carbodiimide-metho-p-toluenesulfonate (0.24 g). After 16 hours at room temperature, the solution was evaporated under reduced pressure and the residue was purified by silica gel chromatography eluting with ethyl acetate. The appropriate fractions gave 38 mg (16.6$) of the title compound as a colorless oil, which is a racemic mixture of the compound of Example 7.

EXAMPLE 14

Methyl- 2, 2- dimethyl- 6- formyl- 1. 3- dioxane- 4- acetate

Cis-2,2-dimethyl-6-(2-phenylethenyl)-1,3-dioxane-4-acetic acid methyl ester (prepared in Example 1) was dissolved in methanol (10 ml) and ozone was passed through the solution at - 78° C, until the solution turned blue. The reaction mixture was purged with nitrogen to remove excess ozone and then dimethyl sulphide was added and the temperature rose to room temperature. The reaction was evaporated in vacuo and the residual oil was purified by chromatography on silica gel eluting with diethyletherhexane (3:1) to provide the title compound.

1-H-NMR (360 MHz, CDCl 3 ) 5:9.53 (1H, s), 4.40-4.23 (2H, m), 3.69 (3H, s), 2.53 (1H, dd, J = 15.8, 7.02 Hz), 2.37 (1H, dd, J = 15.8, 5.98 Hz), 1.85-1.76 (1H, m), 1.44 (3H, s), 1.40 (3H, s), 1.35-1.23 (1H, m).

EXAMPLE 15

3. 3-bis(4-fluorophenyl)-1-bromo-2-(1-methyl-1H-tetrazol-5-yl)-2-propene

A. 5-ethyl-1-methyl-1H-tetrazole

To a slurry of 1,5-dimethyltetrazole (4.9 g, 0.05 mol) in dry tetrahydrofuran (50 mL) was added 2.5 M n-butyllithium in hexanes (20 mL, 0.05 mol) over of 15 minutes at -78°C under an inert atmosphere. The mixture was stirred for 30 minutes, whereby a yellowish precipitate was formed. Methyl iodide (3.7 mL, 0.06 mol) was then added over 15 min. After stirring for an additional 30 minutes, the clear reaction mixture was diluted with water and extracted with ethyl acetate (3 x 50 mL). The aqueous phase was washed with chloroform (2 x 25 mL) and the combined organic phases were dried over sodium sulfate and concentrated under reduced pressure to provide an oil. The oil was purified by distillation to provide 5.2 g (92$) of the title compound, bp: 89-90°C at 0.05 mmHg.

<1->H-NMR (CDCl 3 ) S: 4.05 (s, 3H), 2.86 (q, 2H), 1.41 (t, 3H). <13>C-NMR (CDCl 3 ) S: 156.0, 33.24, 16.75, 11.20.

B. 1. 1-bis(4-fluorophenyl)-2-(1-methyl-1H-tetrazol-5-yl)propanol

To a solution of 5-ethyl-1-methyl-1H-tetrazole (5.6 g, 0.05 mol) (prepared in step A) in 60 mL of dry tetrahydrofuran was added 2.5 M n-butyllithium (20 mL , 0.05 mol) in hexane added over 5 minutes at -78°C (water bath temperature) under an inert atmosphere. The mixture was stirred for 30 min, and a solution of 4,4'-difluorobenzophenone (10.8 g, 0.5 mol) in 25 mL of dry tetrahydrofuran was added over 5 min. The mixture was further stirred for 2 hours while the water bath temperature was slowly heated to -20°C. The reaction was quenched with 1N HCl and extracted with ethyl acetate (3 x 50 mL) and chloroform (3 x 50 mL). The combined organic phases were dried over sodium sulfate and concentrated under reduced pressure to provide a white solid. The solid was purified by crystallization from ethanol-hexane to provide 10.8 g (65%) of the title compound, m.p. 160-161°C.

IR (KBr) v/max: 3400, cm"<1>

<i>H-NMR (CD13) S: 7.8-7.02 (m, 8H), 5.95 (s, 1H), 4.65 (q, 1E), 3.98 (s, 3E) , 1.29 (d, 2E).

<13>C-NMR (CDCl3) S: 162.57, 162.37, 159.14, 156.71, 142.48, 140.54, 128.25, 128.13, 127.52, 127.42 , 114.67, 114.41, 114.38, 78.56, 36.99, 33.43, 33.43, 14.52.

Analysis for C17E16<F>2N4O:

Calculated: C: $61.81, E: $4.88, N: $16.98.

Found: C: 61.79$, E: 4.90$, N: 17.09$.

C. 1. 1-bis(4-fluorophenyl-2-(1-methyl-1H-tetrazole-5-yl)-1-propene

A slurry of 1,1-bis(4-fluorophenyl)-2-(1-methyl-1E-tetrazol-5-yl)-propanol (8.25 g, 0.025 mol) (prepared in step B) and 100 mg p -toluenesulfonic acid, monohydrate in xylene (60 mL) was heated to reflux with a Dean-Stark water collector for 12 hours. The reaction mixture was washed with 1 N NaOH (10 ml) while hot and with water (100 ml). Concentration of the organic phase gave off-white colored crystals of the product. The product was purified by recrystallization from ethanol-hexane to provide 7.1 g (91%) of the title compound as white crystals, m.p. 146-147°C.

IR (KBr) N</>max: 1575; 1500 cm-1.

1 H-NMR (CDCl 3 ) S: 7.42-6.85 (m, 8H), 3.53 (s, 3H), 2.14 (s, 3H).

<13>C-NMR (CDCl3) S: 163.37, 163.08, 160.13, 155.61, 144.60, 145.34, 136.47, 136.42, 136.24, 136.19 , 131.65, 131.54, 131.11, 131.01, 119.53, 115.51, 115.27, 115.22, 33.50, 21.20.

Analysis for C17<H>14<F>2N4:

Calculated: C: 65.37$, H: 4.51$, N: 17.94$.

Found: C: 65.64$, H: 4.61$, N: 18.09$.

D. 3. 3-bis (4-fluorophenyl)-1-bromo-2-(1-methyl-1H-tetrazol-5-yl)-2-propene

A slurry of 1,1.bis(4-fluorophenyl)-2-(1-methyl-1H-tetrazol-5-yl)-1-propene (61.46 g, 0.197 mol) (prepared in step C, N- bromosuccinimide (35.06 g, 0.197 mol) and a catalytic amount of azobis-isobutyronitrile or benzoyl peroxide in carbon tetrachloride (1.2 L) were heated to reflux in an inert atmosphere for 2 hours.The reaction mixture was cooled to room temperature and the solid from the reaction was filtered off. The filtrate was concentrated under reduced pressure and the resulting solid was recrystallized from toluene-hexane to provide 72 g (93%) of the title compound as white crystals, mp 159-160°C.

IR (KBr) v/max: 1600 cm-<1>.

1-H-NMR (CDCl 3 ) S: 7.5-7.1 (m, 8H), 4.44 (s, 2H), 3.53 (s, 3H).

<13>C-NMR (CDCI3) §: 163.94, 163.74, 160.60, 160.45, 143.42, 149.68, 135.20, 135.15, 134.69, 131.31 , 130.90, 130.80, 119.57, 115.94, 115.77, 115.65, 115.50.

Analysis for Ci7H^3F2BrN4:

Calculated: C: 52.19$, H: 3.34$, N: 14.32$.

Found: C: 52.58$, H: 3.47$, N: 14.49$.

EXAMPLE 16

fl, 1-bis(4-fluorophenyl)-2-(1-methyl-1H-tetrazol-5-yl)-1-propen-3-yl-triphenylphosphonium bromide

A slurry of 3,3-bis(4-fluorophenyl)-1-bromo-2-(1-methyl-1H-tetrazol-5-yl)-2-propene (1.95 g, 0.005 mol) (prepared in Example 15, step D) and triphenylphosphine (1.3 g, 0.005 mol) in cyclohexane (25 mL) were heated to reflux. The reaction mixture became a clear solution after 30 minutes and a white precipitate appeared after 1 hour. The mixture was further heated for 8 hours, cooled to room temperature and the solid was collected by filtration and washed with diethyl ether. The white powder was dried in vacuo at 50°C to provide 3.0 g (92%) of the title compound, mp 254-255°C.

IR (KBr) v</>max: 3450, 1600, 1500, 1425 cm-<1>

1 H-NMR (DMSO-d 6 ) S: 7.92-6.80 (m, 23H), 4.94 (6d, 2H), 3.83 (s, 3H).

<13>C-NMR (DMSO-dfc) S: 163.53, 163.36, 160.28, 160.87, 154.04, 153.89, 152.76, 135.11, 134.79, 134 .16, 133.68, 133.54, 130.53, 130.45, 130.35, 130.21, 130.07, 118.02, 116.89, 116.18, 115.89, 115.62 , 115.32, 11.43, 11.39, 34.22, 28.88, 28.22.

Analysis for C35H2sBrF2N4P:

Calculated: C: 64.31$, H: 4.32$, N: 8.57$.

Found: C: 64.02$, H: 4.37$, N: 8.89$.

Claims (6)

1. Tetrazole compound, characterized in that it has the formula (XI) in predominantly cis form, where R1 and R<4> are independently hydrogen, halogen, C1-4 alkyl, C1-4 alkoxy or trifluoromethyl, R 2 , R < 3 >, R < 5 > and r < 6 > are independently hydrogen, halogen, C 1-4 alkyl or C 1-4 alkoxy, R<9> and R<10> are C^_4alkyl, or R<9> and R^-O together with the carbon atom to which they are attached are cyclopentyl, cyclohexyl or cycloheptyl, and r<!2> is hydrogen or a metal cation. 2. Compound according to claim 1, with the formula characterized in that R<9> and R<10> are C^_ 4alkyl, or R<9> and R<1>^ together with the carbon atom to which they are attached are cyclopentyl, cyclohexyl or cycloheptyl, and R<12> is hydrogen or a metal cation. 3. Compound according to claim 2, characterized in that Rg and R<1>^ are methyl, and R12 is hydrogen or a metal cation. 4. Compound according to claim 3, characterized in that R12 is hydrogen. 5. Compound according to claim 2, characterized in that R<9> and R<10>, together with the carbon atom to which they are attached, is cyclohexyl, and R<12> is hydrogen or a metal cation. 6. Compound according to claim 5, characterized in that R1<2> is hydrogen.