FI96602B - Process for the preparation of therapeutically active tetrazole derivatives - Google Patents

Description

- 96602

Process for the preparation of therapeutically active tetrazole derivatives

This invention relates to a process for the preparation of novel inhibitors of the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA). These compounds are useful in the treatment of high blood cholesterol, high blood lipoprotein and atherosclerosis.

Natural fermentation products compactin (R = H) presented by A. Endo, et al. in Journal of Antibiotics, 29, 1346-1348 (1976), and mevinoline (R «CH3), reported by A.W. Alberts, et al. in J.

Proc. Natl. Acad. Sci. U.S.A., 77, 3957 (1980), are highly active agents that lower blood cholesterol, limit cholesterol biosynthesis by inhibiting the enzyme HMG-CoA reductase, a rate-limiting enzyme, and a natural target for the regulation of cholesterogenesis in mammals. Compactin (R = H) and mevinoline (R = CH3, also known as lovastatin) have the structures shown below: chT * H I f 3 N * CH3

’Compactin, R = H

mevinoline, R = CH3

Patents and other publications have also disclosed a number of structurally similar synthetic compounds useful in the treatment of hypercholesterolemia. The synthetic technique most closely related to this is described in the following publication.

U.S. Patent No. 4,198,425, issued April 15, 1980 to S. Mistui et al., Discloses novel Meva-5 lonolkaton derivatives useful in the treatment of hyperlipidemia and having the general formula 15 R 3 wherein A is a direct bond, methylene , ethylene, trimethylene or a vinylene group and R 3, R 4 and R 5 are various substituents.

GP Patent Application 24,348, published March 4, 1981, discloses new blood cholesterol and fat reducing compounds having the structure

HCT

25 I

E

Wherein A is H or methyl; E is a direct bond, -CH 2 -, - (CH 2) 2 -, - (CH 2) 3 - or -CH = CH-; R 1, R 2 and R 3 are each a different substituent, and the corresponding dihydroxy acids obtained by hydrolytic opening of the lactone ring.

35 U.S. Patent No. 4,375,475, issued March 1, 1983 (AK Willard et al.), Discloses substantially the same structures and is consistent with paragraph 24 of the aforementioned EP patent application. With 348.

EP Patent Application 68,038, published January 5, 1983, discloses and claims a resolved transenantiomer having the structure ./tr 10 F i 3 -

15 X

“3 a process for its preparation and its pharmaceutical compositions and the corresponding dihydroxy acid or a pharmaceutically acceptable salt thereof.

International Patent Application WO 84/02131, published June 7, 1984, discloses mevalonolactone analogs having structure 2. Wherein one of R and R 0 is R 5 and the other is primary or secondary C 1-6 alkyl, C 3-6 cycloalkyl or phenyl- (CH 2) n -; X is - (CH 2) n - or -CH 2 CH-; 4 96602 n is 0, 1, 2 or 3; R6

2 is -CH-CH, -C-CH2-COOH and I 2 I

OH OH

R4, R5, R5 · and R6 are various substituents.

International Patent Application WO 84/02903, published August 2, 1984, discloses mevalonolactone analogs having the structures

10 R3 X-2? 5β X-Z

ρθ-φ ,! OCfQ "R ra a1 ra

15 IA IB

(CH2) q- wherein X is - (CH2) n-; n is 0, 1, 2 or 3 and each q is 0 or the other is 0 and the other is 1 and R6

Z - -CH-CH2-C-CH2-COOH and I 2 I 2 OH OH

EP Patent Application 142,146, published May 22, 1985, discloses oxo analogs of blood cholesterol-lowering agents such as mevinoline having the structure

HO

30 0R1 s °

E

z 35 wherein E is -CH 2 -CH 2 -, -CH 2 CH- or - (CH 2) 3 -, and

il? Hit) Η1Ί »1 ·: J

♦ 96602 5

O

E® ° R12

K7 "S

»00 -“ “’ · * CCT

where the dotted line indicates possible double bonds of 0, 1 or 2.

15 In J. Med. Chem., 28, 347-358 (1985) G.E.

Stokker et al. report the preparation and testing of a series of 5-substituted 3,5-dihydroxy-valeric acids and their derivatives.

In J. Med. Chem., 29, 159-169 (1986) W.F.

20 Hoffman et al. show the preparation and testing of a series of 7- (substituted aryl) -3,5-dihydroxy-6-heptenoic acid and their lactone derivatives. One of the preferred compounds in the reported series has structure i

Cl

In J. Med. Chem., 29, 170-181 (1986) G.E.

35 Stokker et al. report the synthesis of the 7- [3,5-disubstituted- (1,1'-diphenyl) -2,5-yl] -3,5-dihydroxy-6-heptenoic acid series and their lactones. Two of the preferred compounds reported in this article have the structures V "cf 7 z * 5

UULc: Xl X.CB

Qg JJ J

C1 CH3 U.S. Patent 4,613,610, issued September 23, 1986 (JR Wareing), discloses pyrazole analogs of mevalonolactone 15 and derivatives thereof useful in the treatment of high blood lipoprotein and atherosclerosis and having the general formula R 6 R 5 -R 7 R 2 R 1 wherein X is - (CH 2) n -, -CH = CH-, -CH = CH-CH 2 - or -CH 2 -CH = CH-; n is 0, 1, 2 or 3 and R 1, R 2, R 3, R 4, R 5, R 6, R 7 and Z are various substituents.

None of said patents or articles discloses or suggests the possibility of preparing the compounds of this invention. The unique structural feature in which the tetrazole moiety is attached to these compounds differs substantially from the prior art referred to above.

• «! L - m t il ii (ltd, -.

966C2 7 The present invention relates to a process for the preparation of novel blood cholesterol lowering compounds of the formula 5 r5 R5 OH OH 0 '1 ^ 1 r2T ^ JJ JL ^ ch R * EJJ .Λ ^ οη3 10 ^ W IIa Rl w * »in which R 1 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; n is 1; and R 7 is hydrogen, a hydrolyzable ester group, or a cation that forms a non-toxic pharmaceutically acceptable salt.

The terms "C 1-4 alkyl", "C 1-6 alkoxycarbonyl" and "C 1-4 alkoxy" as used herein and in the claims (unless otherwise indicated in the context) mean straight-chain or branched alkyl or alkoxy groups such as methyl, ethyl, propyl, isopropyl, butyl , isobutyl, t-butyl, amyl, hexyl, etc. Preferably these. The 25 groups contain 1 to 4 carbon atoms and most preferably contain 1 or 2 carbon atoms. Unless otherwise specified in individual cases, the term "halogen" as used hereinafter and in the claims is intended to include chlorine, fluorine, bromine and iodine, while the term "halogen-30 di" when used hereinafter and in the claims is intended to include chloride, bromide and iodide anions. The term "cation forming a non-toxic pharmaceutically acceptable salt", as used hereinafter and in the claims, is intended to include non-toxic alkali metal salts such as sodium, potassium, calcium and magnesium, ammonium salts and salts of non-toxic amines such as trialkylamines. , pyridine, N-methylmorpholine, N-methylpiperidine and other amines used for salt formation with carboxylic acids 5. Unless otherwise specified, the term "hydrolyzable ester group" hereinafter means and an ester group which is physiologically acceptable and hydrolyzable under physiological conditions, such as C 1-4 alkyl, phenylmethyl and pivaloyloxymethyl ester groups.

In the compounds of formulas IIa and 11b, all double bonds are intended to be in the trans configuration, i.e. (E), as indicated in the structural formulas used herein. These compounds are prepared according to the invention so that a) a compound of formula

Rs r4. ^ JJ-r6 20 R3 JL> s / CH2Or Ιβ r1 V = -n. Wherein R 1, R 2, R 3, R 4, R 5 and R 6 are as defined above, ‘is reacted with either triphenylphosphine to give a phosphonium salt of formula

RS

30 r4 ^^ - R6 \ o11 r2-P IJ I ph 35 vrjr RV = -N If il: »lunnia · = 96602 9 wherein R11 is substituted or unsubstituted phenyl, or with a phosphite to give a phosphonate of formula R3 5 CH2 - P - <0 R10)

r2-P li I

k * J '^ CH3 10 ίΓ V / V = -n ig wherein R10 is C1-4 alkyl; b) reacting one of the products of step a) with an aldehyde of formula OR12 OR12 0

V1 * V'A ' «S

Wherein R 9 is a hydrolyzable ester group and R 12 is t-butyldi-phenylsilyl, in an inert organic solvent in the presence of a strong base; and c) desilylating the product of step b) by reaction with a desilylating agent in an inert organic solvent and in the presence of a small amount of an organic acid to form a compound of formula IIa wherein R 7 is a readily hydrolysable ester group; and if desired d) cleaving the ester functional group R7 by base hydrolysis in an organic solvent to give a compound of formula IIa, wherein R7 is X *, wherein X + is a cation; and if desired e) acidifying the product of step d) to give a compound of formula IIa wherein R 7 is hydrogen; and, if desired, 96602 f) cyclizing the product of formula e) to give a compound of formula Hb by activating the carboxyl radical with carbodiimide in an inert, organic solvent.

The starting materials of formula Ie can be prepared by 5 different methods, preferably using as compounds starting compounds of formula IV

R5 R * —t> 6

10 Γ R

R1 wherein R1, R2, R3, R4, R5 and R6 are as defined above and R6 is hydrogen, C1-4alkoxycarbonyl or methyl. Such methods are described in patent application 880 868.

The process according to the invention for the preparation of compounds of formulas IIa and Hb is illustrated by the following reaction scheme 1.

* | : | »T iilll Evening:.: 966G2 11

Reaction Scheme 1 R5 R «i5“ 4V-R6 5 R11 R3 T c „^ 11 2 Xr11 λΓΝ R2" v ^ il J ^^ CHa 'ÖR' * w * JL ^ CHgBi-

Rs-S Jj X, ch3 ys «ry 3 R = ^ r4-S3" r6

Ie 0 P3 »10 ^^ JL / CH2 — P— <OR10) r2-h li ίΓ

20 Rl W

In Reaction Scheme 1, R 1, R 2, R 1, R 2, R 5 and R 6 are as previously defined. The allyl bromide of formula Ie may be reacted in a conventional manner with phosphines such as triphenylphosphine in an inert organic solvent such as cyclohexane to give a phosphonium salt of formula If wherein R is phenyl which is unsubstituted or substituted by one or two alkyls. - or with a chlorine substituent, and X is bromine, chlorine or iodine. Alternatively, the allyl bromide of formula Ie may be reacted in a conventional manner with phosphites such as trimethyl phosphite or triethyl phosphite either undiluted or in an inert organic solvent, and preferably undiluted to form a phosphonate of formula 3 wherein R ** ® is C - alkyl.

1-4 • r 96602 12

The intermediates of formulas If or Ig can then be converted to the blood cholesterol lowering compounds of formulas Ha and Hb according to the reaction sequence shown in Reaction Scheme 2.

5 Reaction Scheme 2 10 * / CH, - z ·> ti 'ίΓ If or Ig

x2 ~ c I J

tr H-CH.

15 OR12 OR12 O

0 5 Π 20 ** r3 OR12 of2 o

25 R2 — OR9 XII

\ f®3

KM

30 r5 1 RA — R6 0H 0H 0

35 k2— ^ 0r9 X

iA; ch, * w 3 i | IN 4 MM III * »966G2 13

In Reaction Scheme 2, r 1, R 2, R 4, R 1 * and R 6 are g as previously defined, R is a hydrolyzable 1 2 ester group, R is t-butyldiphenylsilyl and O - (R 11 II in © / 11 / T) in 5 Z is -P- (ORX), or -P-Rx X2 ', where R1 is

\ RU

11 C 1-6 alkyl, R is phenyl which is unsubstituted or substituted by one or two substituents R 1, alkyl or chlorine, and X is bromine, chlorine or iodine.

A phosphonium salt of formula If or a phosphonate of formula Ig may be reacted with a silyl-protected aldehyde of formula XI prepared by the methods described in Tetrahedron Letters, 25, 2435 (1984) and also in U.S. Patent No. 15 4. 571,428 to form a silyl-protected compound of formula XII. The reaction may be carried out in an inert organic solvent such as tetrahydrofuran or N, N-dimethylformamide in the presence of a strong base such as lithium diisopropylamide, potassium t-20 butoxide and n-butyllithium at a temperature from about -78 ° C to about 0 ° C. . The compound of formula XII can then be desilylated by well known methods such as 48% hydrofluoric acid and preferably tetrabutylammonium fluoride in an inert organic solvent such as tetrahydrofuran and acetonitrile in the presence of a small amount of organic acid to form the erythro compound of formula X.

The resulting compound of formula X can then be converted to compounds of general formulas IIa and Hb in a conventional manner well known to those skilled in the art.

The compounds of formulas IIa and Hb are competitive inhibitors of the enzyme 3-hydroxy-3-methylglucanoyl-coenzyme A-reductase, a rate-controlling enzyme in cholesterol biosynthesis, and ϋ, << Π9 ✓ Ουυ L 14 therefore, they are selective inhibitors of cholesterol biosynthesis in animals, including man. Therefore, they are useful in the treatment of high blood cholesterol and lipoprotein and atherosclerosis. The biological activity of the compounds of formulas Ha 5 and Hb can be demonstrated by inhibiting cholesterol biosynthesis in rats.

In vivo acute inhibition of cholesterol biosynthesis in rats

Male Wistar rats (160-200 g, placed! -10 na per 2 cages) were maintained on a normal diet (Purina Rat Chow and water, to the extent possible) for at least seven days on an inverted lighting schedule (dark from 7:00 a.m. to 5:00 p.m.). Food was removed 15 hours before dosing. The compounds were administered at 8.00 am by intragastric intubation using 0.5 to 1.0 ml of aqueous or propylene glycol solutions of the sodium salts, lactones or esters of the test compounds. Controls received the same volumes of medium.

Thirty minutes after receiving the test substances, the rats were injected intraperitoneally with 0.9 ml of 0.9% NaCl containing about 120 Ci of sodium [1-14C] acetate (1-3 mCi / mmol) per kg of body weight. After a 60-min joining time, the rats were sacrificed and liver and blood samples were taken. Portions of plasma (1.0 mL) obtained by centrifugation of heparin + EDTA-treated blood and portions of liver homogenates (equivalent to 0.50 g wet weight of liver) were pooled for determination of radiolabeled 3-hydroxysterols. Isolation of sterols with liver samples followed the method of Kates, presented in Techniques in Lipidology, (M. Kates, ed.) Pp. 349, 30 360-363, North Holland Pub. Co., Amsterdam, 1972, while plasma samples were directly saponified and then sterols precipitated with digitonin were isolated. 14C-labeled sterols were quantified by liquid scintillation counting (efficiency corrected). The mean percentage inhibition of 14C incorporation into hepatic and plasma cholesterol was calculated

Il Itt.t IIIII I I 1 St 96602 15 groups and compared with the means of concomitantly treated controls.

Thus, the above experiment provides information on the ability of test substances to suppress cholesterol de Novo 5 biosynthesis in vivo in rats when administered orally. For example, using the above experiment for the compound of Example 6, a 50% inhibitory dose (ED50) was obtained for both plasma and liver cholesterol comparable to that obtained with mevinolin (lovastatin) using a similar method [Alberts, et al., Proc. Natl. . Acad. Sci., 77, 3957-3961 (1980)].

In the following examples, which further illustrate the invention, all temperatures are given in degrees Celsius. Melting points were recorded on a Thomas-Hoover capillary melting point apparatus and boiling points were determined at the pressures given (mmHg) and both temperatures are uncorrected. Proton resonance spectra (1 H-NMR) were recorded on Bruker AM 300, Bruker WM 360 or Varian T-60 CW spectrometers. All spectra were determined in CDCl 3, DMSO-d 6, or D 2 O unless otherwise noted, and chemical shifts are reported as 6 values in the lower field from internal standard tetramethylsilane (TMS), and proton coupling constants are reported in Hertz. (Hz). The distribution methods are denoted as follows: s is a singlet, d is a doublet, t is a trip-25 braid, q is a quartet, m is a multiplet, a broad is a broad peak, and dd is a doublet of a doublet. Carbon-13 nuclear magnetic resonance (13 C-NMR) spectra were recorded on a Bruker AM 300 or Bruker WM 360 spectrometer, and the spectra were broad, with all protons decoded. All spectra were determined in CDCl 3, DMSO-d 6 or D 2 O unless otherwise indicated using an internal deuterium lock, and chemical shifts are reported as δ values toward the lower field from tetramethylsilane. Infrared (IR) spectra were determined on a Ni-colet MX-1 FT spectrometer from 4000 cm-1 to 400 cm-1, the apparatus of which was calibrated on a polystyrene film by absorption at 96602 16 1601 cm-1 and are expressed in inverted centimeters (cm'1). Relative Intensities are reported as follows: s (strong), m (medium), and w (weak).

5 Gas chromatography mass spectra (GC-MS) were determined on a Finnigan 4500 gas chromatograph-quadrupole mass spectrometer with an ionization potential of 70 eV. Mass spectra were also recorded on a Kratos MS-50 using fast atom bombardment (FAB) technology. Mass spectral-10 data are reported in the form: original ion (M *) or protonated ion (M + H) *.

Analytical thin layer chromatography (TLC) was performed on precoated silica gel plates (60F-254) and visualized using UV light, iodine vapors and / or staining with one of the following reagents: a) a 2% solution of phosphomolybdic acid in methanol; b) with reagent a) followed by 2% cobalt sulphate in 5-M H 2 SO 4 with heating. Column chromatography, also referred to as flash column chromatography, was performed on a glass column using 20 fine silica gels (32-63 μm silica gel-H) and pressures slightly above atmospheric pressure and the indicated solvents. All evaporations of the solvents were performed under reduced pressure. As used in this application, the term hexanes means a mixture of isomeric C6 hydrocarbons as specified by American Chemical So-25 ciety, and the term "inert" atmosphere means an argon or nitrogen atmosphere unless otherwise indicated.

Example 1 3,3-Bis- (4-fluorophenyl) -1-bromo-2- (1-methyl-1H-tetrazol-5-yl) -2-propene (Compound Ie) 30 A. 5-ethyl-1- methyl-H-tetrazole

To a slurry of 1,5-dimethyltetrazole (4.9 g, 0.05 mmol) in dry tetrahydrofuran (50 mL) was added a hexane solution of 2,5-n n-butyllithium (20 mL, 0.05 mol) over 15 minutes - At 78 ° C in an inert atmosphere. This mixture was stirred for 30 minutes, during which time i | ·. iti · »n * · ** <4 96602 17 a yellow precipitate formed. Methyl iodide (3.7 mL, 0.06 mol) was then added over 15 minutes. 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 layer was washed with chloroform (2 x 25 mL), and the combined organic layers were dried over sodium sulfate and concentrated in vacuo to give an oily substance, the oil was purified by distillation to give 5.2 g (92%) of the title compound; tr. = 89-90 ° C at 6.67 Pa 10 (0.05 mmHg).

1 H-NMR (CDCl 3) δ: 4.05 (s, 3H), 2.86 (q, 2H), 1.41 (t, 3H) 13 C-NMR (CDCl 3) δ: 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 A] in 60 mL of dry tetrahydrofuran was added 2,5-M n-butyllithium ( 20 mL, 0.05 mol) in hexane over 5 minutes at -78 ° C (bath temperature) under an inert atmosphere. The mixture was stirred for 30 minutes, and a solution of 4,4'-difluorobenzophenone (10.8 g, 0.5 mol) in 25 ml of dry tetrahydrofuran was added over 5 minutes. This mixture was stirred for a further two hours while the bath temperature rose again to -20 ° C. The reaction was quenched with 1 M HCl, and the mixture was extracted with ethyl acetate (3 x 50 mL) and chloroform (3 x 50 mL). The combined organic layers were dried over sodium sulfate and concentrated in vacuo to give a white solid. The solid was purified by crystallization from ethanol / hexane to give 10.8 g • (65%) of the title compound? mp. * 160-161 ° C.

IR (KBr) vBex: 3400 cm-1.

3 H NMR (CDCl 3) δ: 7.8 - 7.02 (m, 8H), 5.95 (s, 1H), 4.65 (q, 1H), 3.98 (s, 3H), 1.29 (d, 2 H).

189 c 6 C 21 C NMR (CDCl 3) δ: 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, 14.52.

Analysis for C 17 H 16 F 2 N 4 O 5 Calculated: C, 61.81; H, 4.88; N, 16.96 Found: C, 61.79; H, 4.90; N, 17.09 C. 1,1-bis- (4-fluorophenyl) -2- (1-methyl-1H-tetrazol-5-yl) -1-propene

A slurry of 1,1-bis- (4-fluorophenyl) -2- (Ι-ΙΟ methyl-1H-tetrazol-5-yl) propanol (8.25 g, 0.025 mol) [prepared in B] and 100 mg p-toluenesulfonic acid monohydrate in xylene (60 mL) was heated at reflux for 12 hours using a Dean & Stark water trap. The reaction mixture was washed with 1 M NaOH (10 mL) while still warm and water (100 mL). Concentration of the organic layer gave the product as off-white crystals. They were purified by recrystallization from ethanol / hexane to give 7.1 g (91%) of the title compound as white crystals; mp = 146-147 ° C.

IR (KBr) 1575; 1500 cm'1.

1 H NMR (CDCl 3) δ: 7.42 - 6.85 (m, 8H), 3.53 (s, 3H), 2.14 (s, 3H); 13 C NMR (CDCl 3) δ: 163.37, 163.08, 160.13, 155.61, 144.60, 145.34, 136.47, 136.42, 136.24, 136.19, 131.65 ,. 25 131.54, 131.11, 131.01, 119.53, 115.51, 115.27, 115.22, 33.50, 21.20.

Analysis for C 17 H 14 N 2 O 2 Calc'd: C, 65.37; H, 4.51; N, 17.94 Found: C, 65.64; H, 4.61; N, 18.09 30 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 C], N-bromosuccinimide (35.06 g, 35 0.197 mol) and a catalytic amount of azobisisobutyronitrile (m-966C219) or benzoyl peroxide in carbon tetrachloride (1.2 liters) were heated to reflux under an inert atmosphere for two hours. The reaction mixture was cooled to ambient temperature and the solid from the reaction mixture was filtered. The filtrate was concentrated under reduced pressure, and the resulting solid was recrystallized from toluene / hexane to give 72 g (93%) of the title compound as white crystals; mp. = 159-160 ° C.

IR (KBr) 1600 cm -1.

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

13 C NMR (CDCl 3) δ: 163.94, 163.74, 160.60, 160.45, 143.42, 149.68, 135.20, 135.15, 134.69, 131.43, 131.31 , 130.90, 130.80, 119.57, 115.94, 115.77, 115.65, 115.50.

15 Analysis for C17H13F2BrN4

Calculated: C, 52.19; H, 3.34; N, 14.32 Found: C, 52.58; H, 3.47; N, 14.49

Example 2 [1,1-bs- (4-fluorophenyl) -2- (1-methyl-1H-tetrazol-5-yl) -1-propen-3-yl] triphenylphosphonium bromide (Compound If)

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 1] and triphenylphosphine 25 (1.3 g, 0.005 mol) in cyclohexane (25 mL) were heated to reflux. The reaction mixture became clear after 30 minutes and a white precipitate appeared after 1 hour. The mixture was heated for an additional eight hours, cooled to ambient temperature, and the solid collected by filtration and washed with diethyl ether. This white powder. dried in vacuo at 50 ° C to give 3.0 g (92%) of the title compound; mp. = 254-255 ° C.

IR (KBr) vBajc: 3450, 1600, 1500, 1425 cm -1.

1 H NMR (DMSO-d 6) δ: 7.92 - 6.80 (m, 23H), 4.94 (6d, 2H), 3.83 (s, 3H); 966 C 213 C NMR (DMSO-d 6) δ: 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, 111.43, 111.39, δ 34.22, 28.88, 28.22.

Analysis for C35H28BrF2N4P

Calculated: C, 64.31; H, 4.32; N, 8.57

Found: C, 64.02; H, 4.37; N, 8.89

Example 3 Methyl- (±) -erythro-9,9-bis- (4-fluorophenyl) -3,5-dihydroxy-8- (1-methyl-1H-tetrazol-5-yl) -6,8-nonadi Enoate (Compound IIa)

To a slurry of phosphonium bromide (0.326 g, 0.5 mmol) [prepared in Example 2] and methyl erythro-3,5-15 bis (diphenyl-t-butylsilyloxy) -6-oxohexanoate [prepared according to the general methods set forth by P.

Kapa, et al. in Tetrahedron Letters, 2435-2438 (1984) and U.S. Patent 4,571,428, issued February 18, 1986 to P.K. Kappa] (0.26 g, 0.4 mmol) in dry dimethylformamide (1 mL), potassium t-butoxide (0.067 g, 0.6 mmol) was added at -20 ° C (bath temperature) under an inert atmosphere. The slurry turned to a red solution and was stirred for 18 hours at -10 ° C. The reaction was completed by the addition of ammonium chloride solution (10 mL) and extraction with methylene chloride (2 x 30 mL). The organic layer was dried over sodium sulfate and concentrated to give an oily substance. the oil was purified by passing it through a pad of silica gel and the main fraction was isolated as an oil (160 mg), the oil (160 mg) was stirred with a solution of 1 M tetra-n-butylammonium fluoride in tetrahydro-furan (2 ml) and a few drops of glacial acetic acid for 18 hours. The reaction mixture was poured into water (10 mL) and extracted with ethyl acetate (3 x 20 mL). The organic layer was dried over sodium sulfate and concentrated to give an oil, the oil was purified by silica gel-35 flash column chromatography eluting with ethyl acetate / hexane. 219 (5602 by mixture (2: 1) to give 0.08 g (75%) of the title compound as an oil MS (CI): m / e - 471 for (M + H) *; 1 H NMR (CDCl 3) Δ: 7.26 - 6.6 (m, 9H), 5.37 (dd, 1H), 4.44 (m, 1H), 4.24 (m, 1H), 3.71 (s, 3H) , 3.56 (s, 3H), 2.47 (d, 2H), 1.58 (m, 2H).

The more polar fraction was also isolated (20 mg) and identified as the corresponding trans-lactone.

Example 4

Dimethyl [3,3-bis- (4-fluorophenyl) -2- (1-methyl-10H-tetrazol-5-yl) -2-propen-1-yl] phosphonate (Compound Ig) A slurry of 3 , 3-bis- (4-fluorophenyl) -1-bromo-2- (1-methyl-1H-tetrazol-5-yl) -2-propene (1.17 g, 3.0 mmol) and trimethyl phosphite (0.41 g , 3.3 mmol), was heated at 100 ° C for 5 minutes. After cooling to ambient temperature, excess trimethyl phosphite was removed in vacuo to give a pale yellow solid. This solid was recrystallized from ethyl acetate / hexane to give the title compound as a pure white solid; mp. = 140-141 ° C.

IR (KBr) 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 (3 H, s), 3.2 (2 H, d)

Analysis for C 19 H 190 N 4 P Calculated: C, 54.29; H, 4.56; N, 13.33 Found: C, 53.83; H, 4.48; N, 13.50 Example 5

Methyl (±) -erythro-9,9-bis (4-fluorophenyl) -3,5-dihydroxy-8- (1-methyl) -lH-tetrazol-5-yl) -6,8-nonadi-enoate (Compound Ha) To a solution of the phosphonate (0.84 g, 2.0 mmol) [prepared in Example 4] was added one equivalent of n-BuLi (2.0 mmol) at -78 ° C (dry ice / acetone). and the resulting deep red solution was stirred at -78 ° C for 15 minutes. Methyl erythro-3,5-bis (diphenyl-t-butylsilyloxy) -6-oxohexanoate [prepared according to the general «22 C fy & r.n ✓ V. ·’ L l.

according to the methods of P. Kapa et al., Tetrahedron Letters, 2435-2438 (1984) and U.S. Patent 4,571,428, issued February 18, 1986 to P. Kapa] (1.30 g). , 2.0 mmol) in THF (2 mL), and the mixture was stirred for 24 h. The reaction mixture was allowed to warm to room temperature during this time. The reaction was quenched with 5 mL of NH.Cl and then extracted with ethyl acetate (2 x 20 mL). The organic layer was dried (Na 2 SO 4) and evaporated under reduced pressure to a yellow oil, the oil was mixed with a 1 M solution of tetra-n-butylammonium fluoride in tetrahydrofuran (4 ml) containing a few drops of glacial acetic acid for 24 hours. The reaction mixture was poured into water (20 mL) and extracted with methylene chloride (3 x 20 mL). The organic layer was dried (Na 2 SO 4) and concentrated, and the oil was purified by silica gel flash column chromatography eluting with ethyl acetate / hexane (2: 1) to give 0.284 g (41%) of the title compound as an oil. MS (CI): m / e for 471 (M + H) + 1 H-NMR (CDCl 3) δ: 7.26 - 6.6 (9H, m), 5.29 (1H, 20 dd), δ , 42 (1H, m), 4.28 (1H, m), 3.69 (3H, s), 3.54 (3H, s), 2.42 (2H, d), 1.5 (2H, m)

Example 6 (conversion of ester Ha where R 7 = C 2 H 5 to salt Ha where R 7 = Na *); Sodium (±) -erythro-9,9-bis- (4-fluorophenyl) -3,5-dihydroxy-8- (1-methyl-1H-tetrazol-5-yl) -6,8-nonadienoate

To a solution of ethyl 9,9-bis (4-fluorophenyl) -3,5-dihydroxy-8- (1-methyl-1H-tetrazol-5-yl) -6,8-no-30 nadieno ( 1.231 g, 2.54 mmol) (preparation shown in application 880 868) in 35 ml of tetrahydrofuran at 0 ° C, 2.54 ml (1.0 equivalent) of 1-M NaOH solution were added dropwise. The rate of addition should be slow enough to prevent the color of the reaction mixture from turning deep gold brown or reddish. The reaction mixture was stirred for 30 minutes.

23

Olis to give a clear homogeneous solution. The reaction mixture was allowed to warm to ambient temperature and the saponification was allowed to continue for another hour. Analytical TLC eluting with 20% MeOH in CHCl 3 (V / V) showed Rf = 0.2 for the desired product. Most of the organic solvent was evaporated under reduced pressure (2.67 kPa, 20 mmHg) at about 10 ° C. The resulting thick syrup was diluted with 4 mL of water, and then the solution was lyophilized at 1.33 Pa (0.01 mmHg) to give 1.126 g (100%) of the title compound as a sodium salt that appears to contain about one mole of water; mp. > 100 ° C, decomposed.

IR (KBr) vBax: 3400 (very broad), 1600 (s), 1575 (s), 1513 (s), 1438 (s), 1404 (s), 1225 (s), 1156 (s), 838 (s) ) cm-1; 1 H-NMR (DMSO-d 6) δ: 7.3-7.4 (4H, m), 7.06 (1H, broad, exchangeable with D 2 O), 7.00-7.06 (2H, m ), 6.87 - 6.91 (2H, m), 6.49 (1H, d, J <15.7 Hz), 5.13 (1H, dd, J »5.4, 15.7 Hz ), 5.05 (1H, broad, exchangeable with D 2 O), 4.14 (1H, m), 3.74 (3H, s), 3.62 (1H, m), 1.99 (1H, dd , J = 3.7, 13.5 Hz), 1.80 (1H, 20 dd, J = 8.5, 13.5 Hz), 1.43 (1H, m), 1.30 (1H, m ) ppm; 13 C NMR (DMSO-d 6) δ: 175.87, 161.85 (d, J 1 = 246, 1 Hz), 161.37 (d, 1 JC.P = 246.9 Hz), 153.08, 144 , 97, 139.88, 136.40, 135.51, 132.22 (d, 3Jc_r = 8.3 Hz), 130.97 (d, 3Jc_r = 8.3 Hz), 124.66, 121.74 , 115.42 (d, 2JC_P-21.9 Hz), 115.12 (d, 2JC_F = 23.4 Hz), 68.23, 65.71, 44.50, 43.55, 33.45 ppm ;

Analysis for C 23 H 21 F 2 N 4 O 4 Na H 2 O Calculated: C, 55.64; H, 4.67; N, 11.28 Found: C, 55.24; H, 4.65; N, 10.85 Example 7 30 (Cyclization of Compound IIa to Lactome Hb)

Trans-6- [4,4-bis (4-fluorophenyl) -3- (l-methyl-lH-tetrazol-5-yl) -1,3-butadienyl] tetrahydro-4-hydroxy-2H-pyran-2 -one A. (±) -erythro-9,9-bis- (4-fluorophenyl) -3,5-dihydro-3-hydroxy-8- (1-methyl-1H-tetrazol-5-yl) -6.8 -nonadienoic acid A solution of ethyl (±) -erythro-9,9-bis- (4-; fluorophenyl) -3,5-dihydroxy-8- (1-methyl-1H-tetrazol-5-240 π 9 ycuu.

yl) -6,8-nonadienoate (0.64 g, 1.32 mmol) in 25 mL of tetrahydrofuran at 0 ° C was treated with 1.32 mL of 1.0 M NaOH solution. The pale yellow suspension was stirred at 0 ° C for two hours to give a clear pale yellow solution. The crude reaction mixture was diluted with 5 mL of aqueous HCl (2-M), and the organics were extracted into ethyl acetate (40 mL x 2). The organic extracts were combined, dried over MgSO 4 and concentrated in vacuo to give a pale yellow gum. The crude dihydroxy acid was dried vigorously under high vacuum (1.33 Pa, at room temperature for 24 hours) before being subjected to the next step.

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

The dry acid from Step A above was dissolved in 100 mL of dry methylene chloride under argon at room temperature, followed by the addition of 1.7 g (4.0 mmol) of 1-cyclohexyl-3- (2-morpholinoethyl) carbodiimide metho-p-t -20 readsulfonate. The lactonization had taken place in less than 15 minutes as shown by analytical TLC (Rf = 0.12) eluting three times with 50% ethyl acetate in hexanes. Most of the organic solvent was evaporated under reduced pressure, and the residue was washed with water (40 ml) and then extracted. 25 with ethyl acetate (40 mL x 2). The organic layers were combined, dried over MgSO 4 and concentrated in vacuo to give 0.54 g (89.7%) of product. A pure sample of the product was obtained by passing it through a thin pad of silica gel eluting with 40% ethyl acetate in hexanes (v / v) to give the title compound which appears to contain about two. si moles of water. MS (CI): m / e) for 1438 (M + H) 1; IR (KBr) vBax: 3425 (broad), 1738 (vs), 1600 (s), 1513 (s), 1225 (vs), 1156 (s), 1038 (s), 838 (s) cm-1? 1 H-NMR (CDCl 3) δ: 7.26 - 7.21 (2H, m), 7.14 (2H, 35 d, J = 8.7 Hz), 6.86 (4H, d, J = 6, 8 Hz), 6.72 (1H, dd, J-0.8, il: IBr IMI Min: i 966G? 15.6 Hz), 5.34 (1H, dd, J-7.1, 15, 6 Hz), 5.18 (1H, m), 4.37 (1H, m), 3.57 (3H, s), 2.68 (1H, dd, J-4.5, 18 Hz), 2 , 60 (1H, ddd, J = 3.63, 2.5, 18 Hz), 2.44 (1H, d, J = 2.6 Hz, variable with D 2 O), 2.00 (1H, dt, J = 18, 1.7 Hz), 1.79 (1H, td, J-2.7, 18 Hz) ppm; 13 C NMR (CDCl 3) δ: 169.20, 163, 162.5, 153.20, 148.81, 135.61, 134.95, 132.45 (d, 3JC_F = 8 Hz), 132.52, 131 , 51, (d, 3JC_p-8 Hz), 130.04, 120.44, 115.95, (d, 2JC_F <21.9 Hz), 115.83 (d, 2JC.P = 21.9 Hz) , 75.67, 62.54, 38.58, 35.58, 10 33.64 ppm:

Analysis for C 23 H 20 F 2 N 4 O 3 2H 2 O Calculated: C, 58.22; H, 5.10; N, 11.81

Found: C, 59.06; H, 4.45; N, 11.25 A sample of the lactone above was crystallized from a cyclohexane-15 benzene mixture to give the title compound as a crystalline solid containing about one mole of benzene; mp. = 105-106 ° C.

Analysis for C 23 H 20 F 2 N 4 O 3 C 6 H 6 Calculated: C, 67.48; H, 5.07; N, 10.85 Found: C, 67.44; H, 5.23; N, 10.59

Example 8 (Conversion of ester 11a where R7 = t-butyl to salt Ha where R7 = O'Na +)

Sodium (±) -erythro-9,9-bis- (4-fluoro-2-methylphenyl) -3,5-dihydroxy-8- (1-methyl-1H-tetrazol-5-yl) -6 , 8- • <nonadienoate

To a solution of t-butyl-9,9-bis- (4-fluoro-2-methylphenyl) -3,5-dihydroxy-8- (1-methyl-1H-tetrazol-5-yl) -6,8- nonadienoate (1.65 g, 3.05 g) [prepared as described in application 880,868] in ethanol (50 mL), sodium hydroxide (3.05 mL, 1-M solution, 3.05 mmol) was added, and the solution is stirred at room temperature for three hours and at 50 ° C for one hour. The solution was concentrated in vacuo to give 1.3 g of the title compound, which appears to contain about one mole of water; mp. = 215-225 ° C (decomposes).

966G2 26

Analysis for C 25 H 25 F 2 N 4 O 4 Na H 2 O Calcd: C, 57.26; H, 5.19; N, 10.69 Found: C, 57.30; H, 5.20; N, 10.00

Example 9 5 (Conversion of ester Ha where R7 = C2H5 to a salt

Ila with R7-Na +)

Sodium (±) - (E), (E), erythro-9- (4-fluorophenyl) -3,5-dihydroxy-8- (l-methyl-lH-tetrazol-5-yl) -9-fenyylinona- 6,8-dienoate hydrate 10 Solution of ethyl (±) - (E), (E) -erythro-9- (4-fluorophenyl) -3,5-dihydroxy-8- (1-methyl-1H-tetrazole -5-yl) -9-phenylnona-6,8-dienoate (160 mg, 0.343 mmol) (prepared in application 880,868) in EtOH (5 mL), treated with 1 M NaOH, (343 μΐ, 0.343 mmol) , and the resulting solution was stirred at 23 ° C for one hour. The solvent was removed by evaporation, and the residue was dissolved in water (2 ml) and lyophilized to give the title compound as a light brown solid (155 mg); mp. 130-137 ° C.

IR (KBr) ν max: 3400 (broad), 1560, 1510 cm -1; 1 H-NMR (DMSO-d 6) δ: 7.50 - 6.80 (m, 9H), 6.51 (d, J = 15.7 Hz, 1H), 5.15 (dd, J = 5, 4 Hz, J '= 15.7 Hz, 1H), 4.15 (m, 1H), 3.70 (s, 3H), 3.65 (broad, 1H), 3.35 (broad, 2H), 1.95 (m, 2H), 1.40 (m, 2H) ppm; 13 C NMR (DMSO-d 6) δ: 176.42, 163.42, 153.17, 146.07, 140 140.03, 139.73, 135.70, 135.64, 132.20, 132.09, 128.72, 128.42, 128.07, 127.98, 124.83, 121.51, 115.51, 115.22, 66.22, 65.69, 44.46, 43.59, 33, 42 ppm.

Analysis for C 23 H 22 FN 4 O 4 Na H 2 O Calcd: C, 57.74; H, 5.06; N, 11.72 Found: C, 58.70; H, 5.10; N, 11.16

Example 10 (Ester IIa where R7 = CH3 is converted to the salt IIa where R7 = Na *)

Sodium (±) - (E) -erythro-9,9-diphenyl-3,5-dihydroxy-3,5-8- (1-methyl-1H-tetrazol-5-yl) nona-6,8-dienoate hydrate glycols; V i t «k <.. U u έ 27

Methyl (E) -9,9-diphenyl-3,5-dihydroxy-8- (1-methyl-1H-tetrazol-5-yl) nona-6,8-dienoate (470 mg, 1.08 mmol) ) (prepared in application 880 868) was dissolved in ethanol (10 mL) and treated with 1 M NaOH (1.08 mL). The reaction mixture was stirred for one hour.

The solvent was evaporated and the residue was lyophilized to give a pale yellow powder (500 mg, 100%); mp. = 145-150 ° C.

IR (KBr) v 3400 (broad), 1610, 1425, 1360 cm -1; 1 H-NMR (DMSO-d 6) δ: 7.60 - 6.60 (m, 10H), 6.52 (d, J = 16 Hz, 1H), 5.12 (dd, J-16 Hz, J '= 5.5 Hz, 1H), 4.20 - 4.05 (m, 1H), 3.80 - 3.55 (m , 1 H), 3.70 (s, 3 H), 3.10 (broad s, 2 H), 2.10 to 1.10 (m, 5 H) ppm;

Analysis for C 23 H 23 N 4 O 4 Na H 2 O Calcd: C, 59.99; H, 5.47; N, 12.17 Found: C, 59.18; H, 5.46; N, 10.96

Example 11 (Conversion of ester Ha with R7 = C2H5 to salt Ha with R7 = Na +) A solution of ethyl (±) - (E) -erythro-9,9-bis- (4-methoxyphenyl) -3 , 5-dihydroxy-8- (1-methyl-1H-tetrazol-5-yl) nona-6,8-dienoate (95 mg, 0.196 mmol) (prepared in application 880 868) in ethanol (15 mL) was treated with 1- M NaOH solution, (196 L), and the mixture was stirred at 23 ° C for one hour. The solvent was removed by evaporation, and the residue was dissolved in water (2 ml) and lyophilized to give the title compound as a brown powder (95 mg, 100%); mp. 175-180 ° C.

IR (KBr) ν max: 3400 (broad), 1600, 1575, 1510 cm-1; 1 H-NMR (DMSO-d 6) δ: 7.70 - 6.65 (m, 9H), 6.55 (d, J = 15.5 Hz, 1H), 5.08 (dd, J = 5, 6 Hz, J '= 15.7 Hz, 1H), 4.14 (broad, 1H), 3.75 (s, 3H), 3.67 (s, 3H), 3.66 (s, 3H), 2.10 to 1.80 (broad, 2H), 1.50 to 1.20 (broad, 2H) ppm; 13 C-NMR (DMSO-d 6) δ: 159.25, 158.80, 153.78, 138.13, 35 132.75, 131.88, 131.60, 131.42, 131.30, 130.41 , 128.68, •

^ V- L

28 128.53, 125.72, 113.74, 113.48, 68.56, 65.89, 55.14, 54.99, 44.68, 43.68, 33.34.

Analysis for C 25 H 27 NaN 4 O 6 2H 2 O Calculated: C, 55.76; H, 5.81; N, 10.41 Found: C, 54.43; H, 5.04; N, 8.15 <”i t I in \ mi 11» »- *

Claims (3)

Arsv ^> s ^ 'CHeBp Ιβ R * 4- «MI r Li 3 R1 w 30 wherein R1, R2, R3, R4, R5 and R6 are as defined above are reacted with either triphenylphosphine to give a phosphonium salt of formula C: <£ ΓΛ «^ <.. · L ο ι 30 R5 R ^ PjJ-R6 5 ^ 3C ^ CHp — Φ ^ -R11 Rij * YY x * u W ^ CH3 r1 V = -n If 10 where R1 is substituted or unsubstituted phenyl, with a tax phosphite to give a phosphonate of formula R 9 R 2 Pjj-R 6 15. IT P p3 II .n V ^^ Jv ^ CHg - P - <OR10) r2 λ— || '1 J)> L ..CH 9 R V = -N Σ 9 20 wherein R 10 is C 1-6 alkyl; b) reacting either of the products of step a) with an aldehyde of formula OR12 OR12 0 25 hyJv ^ v- ^ S) r9 o wherein R9 is a hydrolysable ester group and R12 is t-butyldiphenylsilyl in an inert organic solvent in the presence of a strong base ; and c) desilylating the product of step b) by reaction with a desilylating agent in an inert organic solvent and in the presence of a small amount of an organic acid to form a compound of formula IIa wherein R 7 is a readily hydrolyzable ester group; and if desired • ·, · '1 h 9660? d) cleaving the ester functional group R7 by base hydrolysis in an organic solvent to give a compound of formula Xla, wherein R7 is X *, wherein X * is a cation; and if desired 5 e) acidifying the product of formula d) to give a compound of formula IIa wherein R 7 is hydrogen; and if desired f) cyclizing the product of step e) to give a compound of formula Hb by activating the carboxyl radical 10 with carbodiimide in an inert organic solvent. c./r^ro ^ V- L v ....: 32 For the preparation of therapeutically active tetrazole compounds with the formula 5 1 'R »R4 ± S + |„ R6 R4_P' ^ Ur6? h ° 2TJ T r «R2fc UX CH „<YC 3 Rl II. or R1 and R2 are selected from the group consisting of halogen, C1-4-15 alkyl, C1-4 alkoxy or trifluoromethyl; R 2, R 3, R 5 and R 6 are optionally substituted with halogen, C 1-4 alkyl or C 1-4 alkoxy; n är 1; and R7 is a hydrolyser ester group or a cation having the same or a specific, pharmaceutical moiety, a solid of which is shown in Figure 20). Ie r ^ i "iy 1 pu Rl w 30 color R1, R2, R3, R4, R5 and R6 are defined as having the same properties as the triphenylphosphine for the phosphonium salt form: |: tl * SM: ltl« · 2 • · 0 / ζ <Γ.ο s ^ OL> / R5 r4 "^ 5” r6 5 / CHj— ^ r-Rl1 ^ ίΡΥΥ \ r11 o · '-h, \\ I CH Rl W 10 color R11 with en substituents or substituents on phenyl, or with phosphite on the upper part of the phosphonate with formula pj R4.L? 4j-R6 15. o p3 I II in --P - <OR10) R2-P Il ίΓ RV = -N τ <3 color R10 is C1-4 alkyl; (b) the product of the form of the product a) OR12 0. 25 γΑΛ, »χι o color R9 is a hydrolyser ester group and R12 is t-butyl-diphenylsilyl, i and inert organic solvents in a nerve-free state; and (c) products of the same type (b) desilylers and reactions of the reaction and inert organic solvents and nerves in the form of organic syrups, colors and compounds of the formulation 35 R7 is not a hydrolyzed ester group; and <or - rr o ✓ '.j L <> - £ d) en functional ester group R7 is used in the preparation of hydrolyses from the base and in the form of organic solvents for the production of the formulation Ha, color R7 Mr X *, color X * är en cation; och om sä önskas, 5 (e) products of the same type; (d) surgical agents for the formulation of Ha, color R7 is used; and in this case, f) a product of the process e) cyclizing an active ring of a carboxylic radical with carbodiimides and an inert organ-10 solid solution for use in the formulation of Hb. I; - Itf'i IM II! «'· I