BE1004193A3 - Inhibitors hmg-coa reductase containing phosphorus, intermediate compounds and their use. - Google Patents

Abstract

Inhibitors of structure-responding cholesterol biosynthesis: including their salts, wherein R represents Oh, a lower alkoxy group or a lower alkyl group, Rx represents H or an alkyl group, X represents -O- or -NH-, n is equal to 1 or 2 and Z represents a hydrophobic fixation, intermediate products used in the preparation of these compounds as well as their use.

Description

       

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   "HMG-CoA reductase inhibitors containing phosphorus, intermediates and their use".



   The present invention relates to new phosphorus-containing compounds which inhibit the activity of 3hydroxy-3-methylglutaryl-coenzyme A reductase and which are therefore of interest for inhibiting the biosynthesis of cholesterol, to hypocholesterolemic compositions containing these compounds. , to new intermediate compounds formed during the preparation of the above-mentioned compounds as well as to the use of these compounds for such applications.



   F. M. Singer et al. in Proc. Soc. Exper. Biol.



  Med., 102.370 (1959) and F. H. Hulcher in Arch. Biochem. Biophys. 146,422 (1971) describe that certain mevalonate derivatives inhibit the biosynthesis of cholesterol.



   Endo et al. in U.S. Patent Nos. 4,049,495, 4,137. 322 and 3.983. 140 describe a fermentation product which is active in the inhibition of cholesterol biosynthesis. This product is called compactin and has been reported by Brown et al. [J. Chem. Soc. Perkin 1165 (1976)] as having a complex mevalonolactone structure.



   British Patent No. 1,586,152 describes a group of synthetic compounds corresponding to the formula:
 EMI1.1
 

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 wherein E represents a direct bond, a C 1 -C alkylene bridge, or a vinylene bridge and the different R groups represent a variety of substituents.



   The activity reported in the British patent is less than 1% of that of compactin.



   U.S. Patent No. 4,375,475 to Willard et al. describes hypocholesterolemic and hypolipemic compounds responding to the structure:
 EMI2.1
 
 EMI2.2
 in which A represents H or a methyl group, E represents a direct bond or a group - CH -, - CH-CH .--, - CH-CH-CH.-or-CH = CH-, R <, R, and R3 each represents H, a halogen atom or a C 1 -C alkyl group or a C 1 -C haloalkyl group, phenyl, halogen-substituted phenyl, C 1 -C alkoxy, C-Cc alkanoyioxy, C-alkyl <-C or C-C haloalkyl.

   or a group OR4, in which R4 represents H or a C 2 -C 8 alkanoyl group, benzoyl, phenyl, halophenyl, phenyl-C 1 -C 4 alkyl, C 1 -C 9 alkyl, cinnamyl, C 1 -C haloalkyl C4'allyl, cycloalkyl-C1-C3 alkyl, adamantylalkyl C1-C3 or substituted phenyl-C1-alkyl substituted substituents being chosen from halogen atoms, the groups
 EMI2.3
 C 1 -C 4 alkoxy, C 1 -C 4 alkyl groups or C 1 -C 4 haloalkyl groups as well as the corresponding dihydroxy acids resulting from the hydrolytic opening of the lactone cycle, as well as the pharmaceutically acceptable salts of these acids and C1-C3 alkyl and C1-C3 alkyl esters substituted with phenyl, dimethylamino or acetylamino groups of dihydroxy acids,

   all these compounds

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 constituting the enantiomers having a 4 R configuration in the tetrahydropyran fragment of the trans racemate represented by the above formula.



   Patent application WO 84/02131 (PCT / EP83 / 00308) (based on patent application in the United States of America No. 443,668 of November 22, 1982 and on the patent application in the United States of America No. 548,850 of November 4, 1983) filed in the name of Sandoz
 EMI3.1
 AG describes heterocyclic analogs of mevalono lactone and their derivatives corresponding to the structure:
 EMI3.2
 
 EMI3.3
 / ruz ws in which one of the R and R represents a group:

    < <4 R Sa
 EMI3.4
 and the other represents a primary or secondary C1-C6 alkyl group, C-Couphenyl- (CH) - cycloalkyl, R4 representing hydrogen or a C1-C6 alkyl group, C1-C4 alkoxy (to except t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy, R5 representing hydrogen or a C 1 -C alkyl, C 1 -C alkoxy, trifluoromethyl, fluoro, chloro , phenoxy or benzyloxy, R5a representing hydrogen or a C 1 -C alkyl group, C 1 -C alkoxy, fluorines or chloro and m being equal to 1.2 or 3, provided that R 5 and R 5a both represent hydrogen when R4 represents hydrogen, R represents hydrogen when R 5 represents hydrogen, at most one of R4 and R5 represents trifluoromethyl,

   at most one of R4 and R5 represents phenoxy and at most one of R4 and Rc represents. benzyloxy,

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R2 represents hydrogen or C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy (except t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy,
 EMI4.1
 R3 represents hydrogen or a C 1 -C 4 alkyl group, C 3 alkoxy, fluoro, chloro, phenoxy or benzyloxy, provided that R 3 represents hydrogen when R 2 represents hydrogen, at most l one of R2 and R3 represents trifluoromethyl, at most one of R2 and R3 represents phenoxy and at most one of R2 and R3 represents benzyloxy,
 EMI4.2
 X represents a group- (CH.-) or -CH = CH- (n = 0, 1,

   2 or 3),
 EMI4.3
 R6 5 4 3 \ 2 1 Z represents a group -ra-CH2 - C - CH2-COOH, 1 2 2 OH II
 EMI4.4
 wherein R6 represents hydrogen or a CC alkyl group, in the form of free acid or in the form of a physiologically hydrolyzable and acceptable ester or a lactone thereof or in the form of salt.



   British patent application No. 2 162.179 describes naphthyl analogs of mevalolactone which can be used as inhibitors of cholesterol biosynthesis, responding to the structure:
 EMI4.5
 Z i z 1 1'11 Z in which R. represents a C1-C3 alkyl group,
 EMI4.6
 Z represents a group of formula 21 or 22:

   

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 EMI5.1
 
R7 = H, a hydrolyzable ester group or a cation.
European patent application No. 164. 698 describes the preparation of lactones which can be used as antihypercholesterolemic agents by treatment of an amide with a halide of
 EMI5.2
 organic sulfonyl R SOX and then by separation of the protection group Pr.
 EMI5.3
 
 EMI5.4
 where X = halo, Pr = a carbinol protecting group, 1 R = HouCH, R3, R4 = H, C 1 -C 3 alkyl or phenyl- (C 1 -C 3 alkyl), the phenyl group being optionally substituted by C 1 -C 3 alkyl, C 1 -C 3 alkoxy or halo groups, 2 R = a group of formula (A) or (B):

   

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 EMI6.1
 
 EMI6.2
 6 R = HouOH, R Hou CH-, a, b, c and d = possible double bonds, 7 R = phenyl or benzyloxy, the nucleus in each case being optionally substituted by C 1 -C alkyl groups, or halo, 8 9 R, R = C1-alkyl halo, 3 R5 = C 1 -C 3 alkyl, phenyl or mono- or di- (C1-C3 alkyl) phenyl.



   Anderson, Paul Leroy, published, unexamined published German patent application No. 3,525,256 describes naphthyl analogs of mevalonolactones of the structure:
 EMI6.3
 

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 in which R1 represents an alkyl group, Z = Q, Q1, R7 = H or a hydrolyzable ester group, which can be used as inhibitors of cholesterol biosynthesis and in the treatment of atherosclerosis.



   The patent application WO 8402-903 (based on the patent application in the United States of America No. 460,600 of January 24, 1983) from Sandoz AG describes analogs of mevalonolactones usable as hypolipoproteinemic agents having the structure:
 EMI7.1
 in which the two groups Ro together form a radical of formula:

   
 EMI7.2
 wherein R2 represents hydrogen or C1-C6 alkyl, C1-C6 alkoxy (except t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy and R3 represents hydrogen or a C1-C6 alkyl group, C1-C3 alkoxy, trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy, provided that at most
 EMI7.3
 one of R2 and R3 represents trifluoromethyl, at most one of R2 and R3 represents phenoxy and at most one of R2 and R3 represents benzyloxy, R. represents hydrogen or an alkyl group
 EMI7.4
 in C, -C., iluoro, chloro or benzyloxy, 1 b
R4 represents hydrogen, C 1 -C alkyl, C 1 -C alkoxy (except t-butoxy),

   trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy,

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 EMI8.1
 R5 represents hydrogen or a C1-C6 alkyl group, C1-C6 alkoxy:, trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy,
R5a represents hydrogen or a C 1 -C 6 alkyl, C 1 -C 6 alkoxy, fluoro or chloro group, and provided that at most one of R 4 and R 5 represents trifluoromethyl, at most one of R4 and R5 represents phenoxy and at most one of R4 and R5 represents benzyloxy,
X represents a group:
 EMI8.2
 where n is 0, 1, 2 and 3 and the two q's are 0 or one is 0 and the other is 1,
Z represents a group:

   
 EMI8.3
 
 EMI8.4
 in which R6 represents hydrogen or a CC- alkyl group, with the general condition that -XZ and that the phenyl group bearing R4 are in the ortho position relative to each other, in the form of free acids or in the form of physiologically hydrolysable and acceptable esters or of their # lactones or in the form of salts.
 EMI8.5
 



  European patent application No. 127. 848 (Merck & Co, Inc.) describes derivatives of 3-hydroxy-5-thia-LJ-arylalkanoic acids corresponding to the structural formula:

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 EMI9.1
 in which Z represents:
 EMI9.2
 n is 0, 1 or 2,
E represents a group -CH--, CHL-CH-, -CH2-CH2-CH2-, -CH = CH-CH2- or -CH2-CH = CH-,
R1, R2 and R3 represent, for example, hydro-
 EMI9.3
 gene or group chloro, bromo, fluoro, C 1 alkyl, phenyl, substituted phenyl or OR7 where R7 represents, for example, hydrogen or a C 2 -C 4 canoyl group, benzoyl, phenyl, substituted phenyl , C 1-9 alkyl, cinnamyl, C1-C4 haloalkyl, allyl, CC- cycloalkylalkyl, adamantyl-C 1 -C alkyl,

     or phenyl-alkyl in
 EMI9.4
 13 '& R, RetR represent hydrogen or a chloro, bromo, fluoro or C1-C3 alkyl group, and
X represents, for example, hydrogen, a C1-C3 alkyl group, a cation originating from an alkali metal or from ammonium.



   These compounds have antihypercholesterolemic activity thanks to their property of inhibiting 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase and an antifungal activity.



   French patent application no 2,596. 393 of April 1, 1986 (Sanofi SA) describes 3-carboxy-2- acid derivatives

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 hydroxy-propane-phosphonic, including their salts, which are interesting as hypolipemic agents and which have the formula:
 EMI10.1
 in which R. and R2 = H, a lower alkyl group or optionally a substituted aralkyl group,
R3 and R4 = H, a lower alkyl group or optionally an aralkyl or substituted aryl group.



   These compounds are described as providing greater reductions in cholesterol, triglyceride and phospholipid levels than meglutol.



   European patent application No. 142. 146 (Merck & Co., Inc.) describes mevinoline-type compounds of the structural formula:
 EMI10.2
 in which: Ri represents, for example, hydrogen or a C 1 -C alkyl group,
E represents a group -CH2-CH2, -CH = CH- or - (CH2) r -, and
Z represents:
1)

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 EMI11.1
 oo where X represents a group -O- or -NR ', in which R represents
 EMI11.2
 hydrogen or a C1-C3 alkyl group; R 7 represents a C2-C8 alkyl group and 8 ,, H R represents hydrogen or a CH- group;

   2)
 EMI11.3
 
 EMI11.4
 where R, RetR independently represent, for example, hydrogen, a halogen atom or a C 1 -C 4 alkyl group 4 '3)
 EMI11.5
 
According to the present invention, compounds containing phosphorus are provided which inhibit the enzyme 3-hydroxy-methylgluratylcoenzyme A reductase (HMG-CoA reductase) and which are therefore of interest as hypocholesterolemic agents, these compounds comprising the following fragment:

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 EMI12.1
 in which X represents a group -0- or -NH-, n is equal to 1 or 2 and Z represents a "hydrophobic fixation".



    The expression "hydrophobic fixation" as used in the present case relates to a lipophilic group which, when it is linked to the upper side chain of the HMG type of the molecule by the appropriate linking element ("X" ), attaches to a hydrophobic pocket of the unused enzyme by fixing the HMG CoA substrate, leading to increased potency compared to the compounds in which Z = H.



   In the preferred embodiments, the compounds of the invention correspond to formula 1:
 EMI12.2
 including their salts, formula in which K represents an OH, lower alkoxy or lower alkyl group, Rx represents H or a lower alkyl group, X represents a group -0- or -NH-, n is equal to 1 or 2, Z represents a hydrophobic fixation, as well as their pharmaceutically acceptable salts.



   The terms "salt" and "salts" refer to basic salts formed with inorganic and organic bases. These salts are ammonium salts, alkali metal salts, such as lithium, sodium and potassium salts (which are preferred), alkaline earth metal salts, such as calcium and magnesium salts, salts with organic bases, such as the amine type salts, for example the dicyclohexylamine salt, the benzathine, N-methyl-D-glucamine, hydrabamine salts, the salts with

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 amino acids such as arginine, lysine, etc. Pharmaceutically acceptable, non-toxic salts are advantageous, although other salts are also of interest, for example by isolating or purifying the product.



   Examples of hydrophobic bindings which can be used in the context of the present invention are, without limitation, the following groups:
 EMI13.1
 where the dotted lines represent possible double bonds, for example:
 EMI13.2
 

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 EMI14.1
 
 EMI14.2
 whereR, R, RetR may be the same or different and each represents hydrogen, a halogen atom or a group lower alkyl, haloalkyl, phenyl, substituted phenyl or OR Y, where y R represents H or an alkanoyl group, benzoyl, phenyl, halophenyl, phenyl-lower alkyl, lower alkyl, cinnamyl, haloalkyl, allyl, cycloalkyl-lower alkyl, adamantyl-lower alkyl or substituted phenyl-lower alkyl.



   In the case where Z represents a group:
 EMI14.3
   R - and R - are identical or different and represent hydrogen, a lower alkyl group or OH,
 EMI14.4
 0 6 il R represents a lower alkyl group-C, such

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 EMI15.1
 
 EMI15.2
 or aryl -, R 6a represents a lower alkyl, hydroxy, oxo or halogen group, q is equal to 0, 1, 2 or 3, and R 7 represents H or a lower alkyl group.



  This is how the compounds of formula 1 include:
 EMI15.3
 
 EMI15.4
 The term "lower alkyl" or the term "alkyl" as used alone or as part of another group includes straight and branched chain hydrocarbons containing 1 to 12 carbons in the normal chain, preferably 1 to 7 carbons, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4, 4 -dimethylpentyle, octyl, 2, 2, 4-trimethylpentyle, nonyl, decyle, undecyle, dodecyle, their various branched chain isomers, etc., as well as groups of this type comprising a halo substituent , such as F, Br, Cl or 1 or CF, an alkoxy substituent, an aryl substituent, an alkyl-aryl substituent, a haloaryl substituent, a cycloalkyl substituent,

   an alkyl-cycloalkyl substituent, hydroxy, an alkylamino substituent, an alkanoylamino substituent, an arylcarbonylamino substituent, a nitro substituent, a cyano substituent, a thiol substituent or an alkylthio substituent.

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   The term "cycloalkyl" as used in the present case alone or as part of another group includes saturated cyclic hydrocarbon groups containing 3 to 12 carbons, preferably 3 to 8 carbons, which are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, all of these groups being able to be substituted by 1 or 2 halogens, 1 or 2 lower alkyl groups, 1 or 2 lower alkoxy groups, 1 or 2 hydroxy groups, 1 or 2 alkylamino groups, 1 or 2 alkanoylamino groups, 1 or 2 arylcarbonylamino groups, 1 or 2 amino groups, 1 or 2 nitro groups, 1 or 2 cyano groups, 1 or 2 thiol groups and / or 1 or 2 alkylthio groups.



   The terms "aryl" or "Ar" as used in the present case refer to monocyclic or bicyclic aromatic groups containing 6 to 10 carbons in the cyclic part, such as phenyl, naphthyl, substituted phenyls or naphthyls substituted in which the substituent on phenyl or naphthyl can consist of 1, 2 or 3 lower alkyl, halogen groups (Cl, Br or F), 1, 2 or 3 lower alkoxy groups, 1,2 or 3 hydroxy groups, 1,2 or 3 phenyl groups, 1,2 or 3 alkanoyloxy groups, 1, 2 or 3 benzoyloxy, 1, 2 or 3 haloalkyl groups, 1, 2 or 3 halophenyl groups, 1, 2 or 3 allyl groups, 1,2 or 3 cycloalkylalkyl groups, 1, 2 or 3 adamantylalkyl groups, 1, 2 or 3 alkylamino groups, 1, 2 or 3 alkanoylamino groups, 1, 2 or 3 arylcarbonylamino groups, 1,2 or 3 amino groups,

   1,2 or 3 nitro groups, 1,2 or 3 cyano groups, 1, 2 or 3 thiol groups and / or 1, 2 or 3 alkylthio groups, the aryl group preferably containing 3 substituents.



   The terms "aralkyl", "aryl-alkyl" or "aryl-lower alkyl" as used herein alone or as part of another group refer to lower alkyl groups as discussed above, comprising an aryl substituent, such as benzyl.



   The term "lower alkoxy" and the terms "alkoxy", "aryloxy" or "aralcoxy" as used herein alone or as part of another group include any of lower alkyl, alkyl , aralkyl or aryl mentioned above linked to an oxygen atom.

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   The term "lower alkylthio" and the terms "alkylthio", "arylthio" or "aralkylthio" as used herein alone or as part of another group include any of the lower alkyl, alkyl , aralkyl or aryl mentioned above linked to a sulfur atom.



   The terms "lower alkylamino", "alkylamino", "arylamino", "arylalkylamino" as used herein alone or as part of another group include any of the lower alkyl, alkyl, aryl or aforementioned arylalkyl linked to a nitrogen atom.



   The term "alkanoyl" as used herein as part of another group refers to a lower alkyl group linked to a carbonyl group.



   The terms "halogen" and "halo" as used in the present case refer to chlorine, bromine, fluorine, iodine and CF, chlorine and fluorine being preferred.



   Advantageous compounds are those of formula 1 which correspond to the following structure:
 EMI17.1
 
 EMI17.2
 in which R represents OH, OLi, R represents Li or H, X represents
 EMI17.3
 has 0 or NH, and Z represents a group: o- * ' <RQOJ J k in which RI represents a phenyl group which comprises an alkyl and / or halo substituent or else RI represents a benzyloxy group which comprises a halo substituent, R2 and R are identical and represent a halogen atom or an alkyl group inferior,

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 EMI18.1
 Z can also preferentially represent T; - * a group:

   R csxk Loioi. - 1 ¯? R2
 EMI18.2
 wherein K and m-are as defined above with respect to the compound of formula II. or else Z represents an erour) e:
 EMI18.3
 
 EMI18.4
 in which R represents H, CH3 or OH and R represents a group:
 EMI18.5
 
 EMI18.6
 or phenyl (substituted) methyl, R7 representing H or CH3.



  The compounds of formula 1 of the invention can be prepared according to the following reaction sequence and its description.

  <Desc / Clms Page number 19>

 
 EMI19.1
 



  OSi -) - RP (Oalky j} 1 t osi-C (Cil 3) 3 2 a-Il kyle. L Ili P-CH-CH-CH-Co al 1 b611S 1 2 'S, A - OalkyleC) Si-C (CH-) I-CH2-C-CH2-Co2alkyl Arbuzov reaction (Ra = lower alkyl or C Ces A 6563 lower alkoxy IV
 EMI19.2
 
 EMI19.3
 , GO. - Ra = lower alkyl where Ra represented lower alkyl ai r a VB.-R = OH or R represented lower alkoxy

  <Desc / Clms Page number 20>

 
 EMI20.1
 VB R OH-DCC 0 a'P-CH-5H-CH alkyl (R = OH) (Rb = lower alkyl) R 0-1-CH2 - "- CH2CO2alkyl ') i <& 7 OH = Pyridine f OH ô 1 (Esterification) Si-C (CHJ.



  / "6" 5 6 "5. JVI

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 EMI21.1
 VI 1) (COC1)? CHCl (acid formation Cl) 0 C 81 or 2) Z- (C1I2) n-XH (coupling reaction) R -P-CH-CH-CHCOalkyl & 2 2 = VA. X 0 (C, H,), N, DP Si-C (CH,), Z / \ C6US C6HS 65 6 b VII Rc lower alkyl or lower alkoxy) 1) (n-C4H9) 4NF, CII3COOII, THF (cleavage 0.1 x VII silylated ether) RP-CH-CH-CH-CO R 2) OH-, dioxane (hydrolysis) <"=" 'x OH T' Z I. z

  <Desc / Clms Page number 22>

 
As can be seen in the above reaction sequence, the compounds of formula 1 can be prepared by subjecting iodide A to an Arbuzov reaction by heating the iodide
 EMI22.1
 and phosphonite / phosphite III:

   III R-P (Oalkyl) - where R represents a lower alkyl or lower alkoxy radical, and using standard Arbuzov processes and conditions to form a phosphinate / phosphonate IV:
 EMI22.2
 
Phosphinate / phosphonate IV is a new compound and as such is within the scope of the present invention.



   The phosphinate / phosphonate IV is then subjected to cleavage of the phosphorus ester by treating a solution of compound IV in an inert organic solvent, such as methylene chloride, then with bis (trimethylsily) trifluoroacetamide (BSTFA) and bromide of trimethylsilyl, under an inert atmosphere, such as argon to form phosphinic acid VA, where Ra in compound IV represents a lower alkyl group, that is to say:
 EMI22.3
 u VA lower alkyl-tt¯CH-cli-CE-CO alkyl 2 = 2 2 OH 0 1. Si-C (CHg) g C6Hg C65

  <Desc / Clms Page number 23>

 or phosphonic acid VB (in which Ra in compound IV represents a lower alkoxy group), that is to say:
 EMI23.1
 
Compounds VA and VB are new intermediate compounds and as such they fall within the scope of the present invention.



   When the phosphonic acid VB is obtained, it is esterified by treating the acid VB in dry pyridine with an alcohol: VC ROH (whereR represents a lower alkyl) and dicyclohexyl carbodiimide and the mixture is stirred of reaction resulting under an inert atmosphere, such as argon, to form a mono phosphonic alkyl ester VI:
 EMI23.2
 
The ester VI or phosphinic acid VA is then dissolved in an inert organic solvent, such as methylene chloride, benzene or tetrahydrofuran (THF) and treated with trimethylsilyldiethylamine and stirred under an inert atmosphere , such as argon; the mixture is evaporated and then dissolved in methylene chloride (or in any other suitable inert organic solvent).

   The resulting solution is cooled to a temperature of the order of about 0 ° C. to about 25 ° C., it is treated with oxalyl chloride and then it is concentrated by evaporation to give

  <Desc / Clms Page number 24>

 crude phosphonochloridate.

   The phosphonochloridate is dissolved in an inert organic solvent, such as methylene chloride, benzene, pyridine or THF, the solution is cooled to a temperature of about -20 C to about 0 C and is treated by
 EMI24.1
 using a molar ratio of VI or VA / B of the order of about 0.511 to about 3/1, and preferably from about 1/1 to about 2/1, and then with triethylamine and catalytic 4-dimethylaminopyridine (DMAP) to form the adduct VII:
 EMI24.2
 wherein R represents a lower alkyl or lower alkoxy group.



   Compound VII is subjected to cleavage of the silylated ether by treating compound VII in an inert organic solvent, such as tetrahydrofuran, with glacial acetic acid and tetrabutylammonium fluoride to form the ester VIII:
 EMI24.3
 
Ester VIII can then be hydrolyzed to an alkali metal salt or the corresponding acid, that is to say when Rx represents an alkali metal or H, by treatment with a strong base, such as lithium hydroxide in the presence of dioxane, tetrahydrofuran or another inert organic solvent, under

  <Desc / Clms Page number 25>

 an inert atmosphere, such as argon, at 25 ° C., using a base / ester VIII molar ratio of the order of about 1/1 to about 1.1: 1, to form the corresponding alkali metal salt:

   
 EMI25.1
 wherein R represents a lower alkyl or lower alkoxy group.



   We can then treat compound VIIIA with a strong acid, such as HCl to form the corresponding acid VIIIB:
 EMI25.2
 
Ester VIII, in which R represents a lower alkoxy group, can be converted to the corresponding dialkali metal salt by treating ester VIII with a strong base at 50-60OC and using a base / ester VIII molar ratio of the order from about 2/1 to about 4/1 to form the product of formula VIIIC:
 EMI25.3
 0 Il VIIIC alkali metal O-P-CH-gH-CH - alkali metal 1 2 = H2-Co2m X OH 1 (CH 2) n (CH2) &verbar; 2n z
 EMI25.4
 The dialkali metal salt VIIIC can be converted to the corresponding acid, in which R represents OH, by treatment with a strong acid, such as HC1 to form the compound VIIID:

   

  <Desc / Clms Page number 26>

 
 EMI26.1
 
The starting iodide A can be prepared starting from the bromide C:
 EMI26.2
 [prepared using the methods as described in Tetrahedron Lett. 26, 2951 (1983)]
 EMI26.3
 which is brought into solution in dimethylformamide (DMF) with imidazole and 4-dimethylamino pyridine and the resulting solution is treated with t-butyldiphenyl silyl chloride under an inert atmosphere, such as l 'argon to form silyl ether D:
 EMI26.4
 
A solution of silylated ether D in an inert organic solvent, such as methyl ethyl ketone or DMF is treated with sodium iodide under an inert atmosphere, such as argon, to form iodide A.



   The starting compound B can be prepared:
 EMI26.5
 as described below according to the definition of Z and X.



   This is how the compounds of formula B, in which Z represents:

  <Desc / Clms Page number 27>

 
 EMI27.1
 and X represents 0, that is to say the compounds of the structure:
 EMI27.2
 can be prepared by treating aldehyde E:
 EMI27.3
 by a reducing agent, such as lithium aluminum hydride or sodium borohydride.



   The compounds of formula B, in which Z represents:
 EMI27.4
 and X represents N, that is to say the compounds of the structure:
 EMI27.5
 

  <Desc / Clms Page number 28>

 can be prepared by oxidation of aldehyde E by treating E in solution with acetone using, for example, a Jones reagent to form acid F:
 EMI28.1
 which, in suspension with methylene chloride, is treated with oxalyl chloride to form the corresponding acid chloride, which is dissolved in an inert organic solvent, such as tetrahydrofuran, and treated with a mixture of hydroxide ammonium concentrated in tetrahydrofuran to form an amide of the structure:

   
 EMI28.2
 Amide G is then reduced to the corresponding amide B2 by treating
 EMI28.3
 G by a reducing agent, such as lithium aluminum hydride.



  The starting compounds of formula B, in which Z represents:
 EMI28.4
 
 EMI28.5
 and X represents 0 or -NH-, that is to say the compounds of the structure:

  <Desc / Clms Page number 29>

 
 EMI29.1
 in which X represents 0, are described by C. H. Heathcock et al., J. Org. Chem. 50.1190 (1985). The compounds of formula H ', in which X represents NH, can be prepared by the reductive amination of:
 EMI29.2
 (prepared as described by C. H. Heathcock et al., supra) by treating J with ammonium acetate and sodium cyanoborohydride in the presence of an alcoholic solvent, such as methanol.



   The starting compounds of formula lob in which Z represents:
 EMI29.3
 and X represents 0, that is to say the compounds of the structure:

  <Desc / Clms Page number 30>

 
 EMI30.1
 are described in patent application WO 8402-903 and in British patent application No. 2 162.179, both filed in the name of Sandoz.



   The starting compounds of formula B, in which Z represents:
 EMI30.2
 and X represents NH, that is to say the compounds of the structure:
 EMI30.3
 
 EMI30.4
 1 1 can be prepared by reductive lamination of aldehyde Q:
 EMI30.5
 by treating Q with ammonium acetate and sodium cyanoborohydride in the presence of an alcoholic solvent, such as methanol.



   The compounds of the invention can be prepared: in the form of racemic mixtures and can then be resolved: to obtain the S isomer, which is preferred. However, the compounds: of the invention can be prepared directly in the form of dE

  <Desc / Clms Page number 31>

 their S isomers, as described in the present case as well as in the processing examples given below.



   The compounds of the invention are inhibitors of 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase and are therefore of interest in the inhibition of cholesterol biosynthesis, as demonstrated by the following tests.



  1) Rat HMG-Coa reductase
The activity of rat HMG-CoA reductase is measured using a variant of the method described by Edwards (Edwards, P. A., et al., J. Lipid Res. 20 / 40,1979). Rat liver microsomes are used as the enzyme source, and enzyme activity is determined by measuring the conversion of the 14C-HMG-CoA substrate to C-mevalonic acid. at. Preparation of microsomes
The livers of 2-4 decapitated Sprague Dawley rats fed with cholestyramine are separated and homogenized in phosphate buffer A (potassium phosphate: 0.04 M, pH 7.2; KC1: 0.05 M; sucrose: 0.1 M; EDTA: 0.03 M; aprotinin: 500 KI units / ml).

   The homogenization product is rotated at 16,000 x g for 15 minutes at 4 C. The supernatant is separated and recentrifuge under the same conditions a second time. The second supernatant is spun from 16,000 x g to 100,000 x g for 70 minutes
 EMI31.1
 at 4 C. The microsome pellets are resuspended in a minimum volume of buffer A (3-5 ml per liver) and homogenized in a glass / glass homogenizer. Dithiothreitol (10 mM) is added, the preparation is divided into aliquots, they are quickly frozen in acetone / dry ice and stored at -80 C.



  The specific activity of the first microsomal preparation was 0.68 nmole of mevalonic acid / mg protein / minute. b. Enzyme determination
The reductase is assayed in an amount of 0.25 ml containing the following components at the final concentrations indicated:

  <Desc / Clms Page number 32>

 
 EMI32.1
 0.04 M Potassium phosphate, pH 7.0 0.05 M KC1 0.10 M Sucrose
0.03M EDTA
0.01M Dithiothreitol
3.5 nM NaCI
1% Dimethyl sulfoxide 50-200 #g Microsomal protein
100 uM C- [DL] HMG-CoA (0.05! LCi, 30-60 mCi / mmole)
2.7 mM NADPH (nicotinamide adenine dinucleotide phosphate) The reaction mixtures are incubated at 37 C. Under the conditions described,

   the enzymatic activity increases linearly up to 300 μg of microsomal protein per reaction mixture, and is linear with respect to the incubation time up to 30 minutes. The standard incubation time chosen for drug studies is 20 minutes, which leads to a 12-15% conversion of the HMG-CoA substrate to mevalonic acid. The substrate [DL] HMG-CoA is used at 100 pM, twice the concentration necessary to saturate the enzyme under the conditions described. NADPH is used in excess concentration, at 2.7 times the concentration required to obtain the maximum enzymatic speed.



   The standardized assays for evaluating the inhibitors are carried out according to the following process. The microsomal enzyme is incubated in the presence of NADPH at 37 ° C. for 15 minutes. DMSO with or without test compound is added as a vehicle, and the mixture is further incubated for 15 minutes at 37 C.
 EMI32.2
 14 the enzymatic assay by the addition of substrate C-HMG-CoA. After an incubation of 20 minutes at 37 ° C., the reaction is stopped by the addition of 25! l1 of 33% KOH. 3Hmevalonic acid (0.05 uCi) is added and the reaction mixture is left to stand at room temperature for 30 minutes. 50 μl of 5N HCl is added to lactonize the mevalonic acid.

   Bromophenol blue is added as a pH indicator to control an adequate pH drop. The lactonization is allowed to continue for 30 minutes

  <Desc / Clms Page number 33>

 at room temperature. The reaction mixtures are centrifuged for 15 minutes at 2800 rpm. The supernatants are placed in layers on 2 grams of AG 1-X8 anion exchange resin (Biorad, in the form of formate) introduced into 0.7 cm glass columns and eluted with 2.0 ml of H 2 O. The first 0.5 ml are discarded and the next 1.5 ml is harvested and counted for both tritium and carbon-14 in 10.0 ml of Opti-fluorine scintillation fluid.

   The results are calculated in the form of nmoles of mevalonic acid produced during a period of 20 minutes, and are corrected to 100% recovery of tritium. The effects of the drugs are expressed in the form of I50 values (concentration of drug producing 50% inhibition of the enzymatic activity) from the response curves to the overall doses with the 95% confidence interval indicated.



   The conversion of drugs in the form of lactone to their sodium salts is carried out by dissolving the lactone in DMSO, adding a 10-fold molar excess of NaOH, and leaving the mixture to stand at room temperature for 15 minutes. The mixture is then partially neutralized (pH 7.5-5.0) using IN HCl, and diluted in the enzyme reaction mixture.



  2) Synthesis of chdesterot in freshly isolated rat hepatocytes
Compounds which exhibit activity as HMG-CoA reductase inhibitors are examined for their ability to inhibit the incorporation of C-acetate into cholesterol from suspensions of freshly isolated rat hepatocytes using the methods initially described by Capuzzi et al. (Capuzzi, D. M. and Margolis, S., Lipids, 6: 602.1971). at. Isolation of rat hepatocytes
Sprague Dawley rats (180-220 grams) are anesthetized with Nembutol (50 mg / kg). The abdomen is opened and the first branch of the portal vein is ligated. Heparin (100-200 units) is injected directly into the abdominal vena cava.

   A single closing suture is made on the distal section of the

  <Desc / Clms Page number 34>

 portal vein, and a cannula is placed in the portal vein between the suture and the first branching vein. The liver is perfused at a rate of 20 ml / minute with an oxygenated buffer A (HBSS without calcium or magnesium containing 0.5 mM EDTA) preheated (37 C) after having cut the vena cava to allow the flow of effluent.



  The liver is also perfused with 200 ml of preheated buffer B (HBSS containing 0.05% bacterial collagenase). After the infusion with buffer B, the liver is excised and decapsulated in 60 ml of Waymouth medium, thus allowing free cells to disperse in the medium. The hepatocytes are isolated by centrifugation at slow speed for 3 minutes at 50 x g at room temperature. The hepatocyte pellets are washed once in Waymouth medium, counted and assayed for their viability by the exclusion of trypan blue. These cell suspensions enriched in hepatocytes generally show a viability of 70-90%. b.

   Incorporation of C-acetate into cholesterol
The hepatocytes are resuspended at a rate of 5 × 10 6 cells per 2.0 ml in an incubation medium (MI) [0.02 M
 EMI34.1
 Tris-HCl (pH 7.4), 0.1 M KC1, 3.3 mM sodium citrate, 6.7 mM nicotinamide, 0.23 nM NADP, 1.7 mM glucose-6 - phosphate].



   The test compounds are generally dissolved in DMSO or DMSO / H2O 0/3) and added to the incubation medium. The final DMSO concentration in MI is less than or equal to 1.0% and has no significant effect on the synthesis of cholesterol.



   Incubation is started by adding C-acetate (58 mCi / mmol; 2 uCi / ml) and placing the cell suspensions (2.0 ml) in 35 mm tissue culture dishes, at 37 C for 2 0 hours. After incubation, the cell suspensions are transferred to glass centrifuge tubes and spun at 50 x g for 3 minutes at room temperature. The cell pellets are resuspended and dissolved in 1.0 ml of H-C and placed in an ice bath.



   The lipids are extracted essentially as described by Bligh E. G. and W. J. Dyer, Can. J. Biochem. and Physiol.,

  <Desc / Clms Page number 35>

 37; 911, 1959. The lower organic phase is separated and dried under a stream of nitrogen, and the residue is resuspended in chloroform / methanol (2/1) (100 µl). The total sample is distributed in drops on thin layers of silica gel (LK6D) and developed in a mixture of hexane / ethyl ether / acetic acid (75/25/1). The plates are scanned and counted using an BioScan automatic scanning system. The radioactive labeling of the cholesterol peak (RF 0.28) is determined and is expressed in total counts per peak and as a percentage of the labeling of the total lipid extract.

   Cholesterol peaks in control cultures usually contain 800-1000 cpm and represent 9-20% of the labeling present in the total lipid extract; results compatible with Capuzzi et al., indicating 9% of labeling extracted in cholesterol.



   The effects of the drugs (percentage inhibition of cholesterol synthesis) are determined by comparing the percentage of labeling in cholesterol for the control cultures and the cultures treated with the drugs. Dose response curves are constructed from overall results from two or more studies, and the results are expressed as I50 values with a 95% confidence interval.



  3) Synthesis of cholesterol in human skin fibroblasts
A compound selectively promoting greater inhibitory activity in liver tissue would be a property for an inhibitor of cholesterol synthesis. Therefore, in addition to evaluating inhibitors of cholesterol synthesis in hepatocytes these compounds are also tested for their activity as inhibitors of cholesterol synthesis in cultured fibroblasts. a) Cultures of fibroblasts of human skin
Human skin fibroblasts are grown (passage 7-27) in an Eagle's minimum essential medium (EM) containing 10% fetal calf serum.

   For each experiment, stock cultures are trypsonized to disperse the single-cell layer, counted and spread in 35 mm tissue culture wells (5 x 105 cells / 2.0 ml). Cultures are incubated for 18 hours

  <Desc / Clms Page number 36>

 at 37 C in a mixture of 5% C02 and 95% humidified ambient air. Biosynthetic enzymes of cholesterol are induced by separating the medium containing serum, by washing the single-cell layers, by adding 1.0 ml of ME containing 1.0% of bovine serum albumin without fatty acids and by incubating the cultures for 24 additional hours. b. Incorporation of C-acetate into cholesterol
Induced fibroblast cultures are washed with EMEM 100 (Earle minimal essential medium).

   The compounds are dissolved
 EMI36.1
 test in DMSO or DMSO / EM (1/3) (final DMSO concentration in cell cultures less than or equal to 1.0%), they are added to the cultures and the cultures are incubated beforehand for 30 minutes at 37 C in a mixture of 5% CO / 93 96 of humidified ambient air. After preincubation with the drugs, [1-14C] Na acetate (2.0 uCi / ml; 58 mCi / mmol) is added, and the cultures are reincubated for 4 hours. After the incubation, the culture medium is separated and the single-cell layer (200 μg of cellular protein per culture) is rubbed in 1.0 ml of H2O. The lipids of the destroyed cell suspension are extracted into a mixture of chloroform and methanol, as described for the hepatocyte suspensions.

   The organic phase is dried under nitrogen and the residue is resuspended in a mixture of chloroform and methanol (2/1) (100 μl) and then the total sample is distributed in drops on thin layer plates silica gel (LK6D) and analyzed as described for hepatocytes.



   The inhibition of cholesterol synthesis is determined by comparing the percentage of labeling in the cholesterol peak from control cultures and cultures treated with the drugs. The results are expressed in the form of I50 values and are obtained from curves of response to overall doses from two or more experiments. The 95% confidence interval for the I50 value is also calculated from the overall dose response curves.



   Another aspect of the present invention consists of a pharmaceutical composition formed from at least one of the compounds

  <Desc / Clms Page number 37>

 of formula I in combination with a pharmaceutical carrier or diluent. The pharmaceutical composition can be formulated using conventional solid or liquid carriers or diluents and pharmaceutical additives of a type suitable for the desired mode of administration. The compounds can be administered orally, for example in the form of tablets, capsules, granules or powders, or they can be administered parenterally in the form of injections, these dosage forms containing 1 to 200 mg of active compound per dosage, when used during treatment.

   The dose to be administered depends on the unit dose, symptoms, and the patient's age and body weight.



   The compounds of formula I can be administered in a similar manner to known compounds usable for inhibiting the biosynthesis of cholesterol, such as lovastatin, to mammalian species, such as humans, dogs, cats, etc. Thus, the compounds of the invention can be administered at a rate of approximately 4 to 2000 mg in a single dose or in the form of individual doses from 1 to 4 times per day, preferably at a rate of 4 to 200 mg in divided doses of 1 to 100 mg, more advantageously at a rate of 0.5 to 50 mg 2 to 4 times a day or in a delayed-release form.



   The following Processing Examples represent preferred embodiments of the present invention. Unless otherwise indicated, all temperatures are expressed in degrees Celsius. Rapid chromatography is carried out either on Merck 60 silica gel or on Whatmann LPS-I silica gel.



  Reverse phase chromatography is performed on a CHP-20 MCI gel resin manufactured by Mitsubishi Ltd.



   Example 1 Monolithic acid salt (S) -4- [) l4'-fluoro-3, 3 ', 5-trimethyl [l, l'-biphenyl] -
 EMI37.1
 2-yl] methoxy] methoxyphosphinyl] -3-hydroxybutanol'que A. N- (2,4-Dimethylbenzylidene) benzeneamine
Merck reference, United States patent No. 4,375,475, page 39.

  <Desc / Clms Page number 38>

 a solution of z-4-dimetnylbenzaldenyde straight distilled (Aldrich; 6.97 ml, 50 mmol) and distilled aniline (Aldrich; 4.56 ml; 50 mmol) in dry toluene (80.0 ml) is heated at reflux for 3.0 hours under argon in a flask fitted with a Dean-Stark apparatus. The mixture is cooled and then concentrated by evaporation in vacuo to a yellow oil.

   The crude oil is purified by Kugelrohr distillation (0.5 mm Hg; 160-180 C) to obtain 8.172 g (78.1%) of benzeneimine, cited in the title, desired in the form of a light yellow oil , which crystallizes by abandor
 EMI38.1
 into a low melting point solid. Chromatography on thin layer: hexane-acetone (4/1), RF = 0.67 and 0.77 (geometric isomers)
 EMI38.2
 U. V. eut 12 * B. N 1 4. 1 1 "Y" o CHg 2 1 CH3 12, Reference: Merck, United States Patent No. 4,375,475 page 39.



   A mixture of benzeneimine from part A (6.0 g 28.7 mmol) in glacial HOAc (144 ml) is treated with palladium (II) acetate (6.44 g; 28.7 mmol) and the homogeneous light red solution is heated at reflux under argon for 1 hour. We filtn
 EMI38.3
 hot, the resulting cloudy mixture through a 12.7 mm Celite bed in 900 ml of HO. The precipitated orange solid is collected by filtration and dried under vacuum at 650C on P 205 for 16.0 hours to give 10.627 g (85.5%) of rubricated palladium complex, desired, in the form of an orange solid with a melting point of 194 -196 C. (Melting point in the literature of a recrystallized analytical sample = 203 -205 C).

  <Desc / Clms Page number 39>

 



  C. 4'-Fluoro-3, 3 ', 5-trimethyifl, 1'-biphenyl] -2-carboxaldehyde (1) Bromo 4-fluoro-3-methylphenyl] magnesium Reference: Merck, patent of the United States of America n 4. 375. 475, pages 37 and 38.



   The Grignard reagent from part C (l) is prepared under the heading by adding dropwise 5-bromo-2-fluorotoluene (22.5 g; 60.9 mmol; Fairfield Chemical Co.) at a rate sufficient to maintain the reaction at reflux temperature to magnesium turns with stirring (1.35 g; 55.4 mmol; 8.0 equivalents) in dry Et 20 (70.0 ml). The reaction is started in an ultrasonic device. After the addition of bromide has been carried out, the mixture is stirred for one hour under argon at room temperature, it is heated under reflux for 15 minutes and then it is allowed to cool to room temperature.



  (2) 4'-Fluoro-3,3 ', 5-trimethyl [1,1'-biphenyl] -2-carboxaldehyde
In a second container, a mixture of the dipalladiated complex from part B (3.0 g; 6.92 mmol) and triphenylphosphine (14.52 g; 55.4 mmol) is stirred at room temperature under argon for 30 minutes; 8.0 equivalents) in dry benzene (100 ml).



  The Grignard reagent from part C (1), freshly prepared and filtered (glass wool plug) is then added in a single portion using a cannula to this solution and the mixture is stirred for 1.5 hours at room temperature under argon. 6.0 N HCl (35 ml) is added, the mixture is stirred for an additional hour at room temperature, and then filtered through packed Celite (12.7 mm bed). The filtrate is extracted with Et20 (250 ml), the extract is washed with bromine (2 x 100 ml), dried over anhydrous MUS04 and concentrated by evaporation in vacuo to obtain 13.35 g of a viscous orange oil, which crystallizes by abandonment.



  The crude orange solid is purified by means of rapid chromatography on silica gel (700 g), eluting with hexane and then with a mixture of hexane and Et2O (95/5). The product fractions are concentrated by evaporation to obtain 1.507 g (89.9%) of the aldehyde mentioned in the title, desired in the form of a light yellow solid with a melting point of 72O-75OC. (The literature reports

  <Desc / Clms Page number 40>

 
 EMI40.1
 a melting point of 73 -74 C). Hex-Et20 (95/5) thin layer chromatography, Rf = 0, 40, U. V. and PMA.



  D. 4'-Fluoro-3, 3 ', 5-trimethyl [1J'-biphenyl] -2-methanol
A cooled solution (0 C; dried Et20 ice bath (15.0 ml) is treated with LiAlH. (239 mg; 6.82 mmol; 0, equivalent) and the gray suspension is drip dropwise on a 15 minute period with a solution of the aldehyde from Part C (3.0 g; 12.4 mmol) in dried Et20 (15 ml) The mixture is stirred at room temperature under argon for 30 minutes : it is brought back to 00C and cooled by additions drop by drop
 EMI40.2
 successive 260 µl HO, 260 µl 15% NaOH and 780 H2O.

   The suspension is diluted with EtOAc, filtered through anhydrous NaSO over packed Celite (12.7 mm bed) and the colorless filtrate is concentrated by evaporation under vacuum to obtain 2.99 g (98, 8%) of a white solid. Trituration of the crude solid pa of cold hexane and drying under vacuum makes it possible to obtain 2.467 (81.6%) of the alcohol mentioned in the heading, desired in the form of a white solid with a melting point of 102-103 C. Thin layer chromatography: Hex-EtOAc (9/1), Rf = 0.24, UV and PMA.
 EMI40.3
 



  E. (S) -3- acid methyl ester! I (1, 1-dimethylethyl) diphenylsilyl] oxy] -4- (hydroxymethoxyphosphinyl) butanolque (1) Methyl ester of (S) -4-bromo-3-hydroxybutanoic acid (1) (a) Calcium salt hydrate d 'acid [R- (R *, R *)] -2, 3, 4-trihydroxy butanoic Reference: Carbohydrate Research 72, pages 301-304 (1979).



   Calcium carbonate (50 g) is added to a solution of D-isoascorbic acid (44.0 g; 250 mmol) in H2O (625 ml
 EMI40.4
 the solution is cooled to 00C (ice bath) and treated by portion with 30% HO-(100 ml). The mixture is stirred at 30 -40 ° C. (oil bath) for 30 minutes. Darco (10 g) is added and the black suspension is heated in a water bath until all release of O2 stops.

   The suspension is filtered through Celite and concentrated by evaporation under vacuum (bath temperature of 40 ° C. The residue is taken up in H2O (50 ml), it is heated in a water bath and CH30H is added until the solution becomes cloudy

  <Desc / Clms Page number 41>

 The gummy precipitous solid is collected by filtration and dried in air to obtain 30.836 g (75.2%) of the desired calcium salt in the form of a white powdery solid. Thin layer chromatography: iPrOH-NH. OH-HO (7/2/1), Rf = 0.19, PMA.



   (1) (b) [S- (R *, S *)] -2, 4-dibromo-3-hydroxybutanoic acid methyl ester Reference: Bock, K. et al., Acta Scandinavica (B) 37 , pages 341-344 (1983).



   The calcium salt from part (1) (a) (30 g) is dissolved in 30-32% HBr in acetic acid (210 ml) and stirred at room temperature for 24 hours. Methanol (990 ml) is then added to the brown solution and stirred overnight. The mixture is concentrated by evaporation in an orange oil, it is taken up in CH30H (75 ml), it is heated under reflux for 2.0 hours and it is concentrated by evaporation. We distribute the residue
 EMI41.1
 between EtOAc (100 ml) and H2O, the organic phase is washed with 1H20 (2 x) and brine, then dried over Na-SO. anhydrous and concentrated by evaporation to give 22.83 g (90.5%) of crude dibromide in the form of a light orange oil.



  Thin layer chromatography: EtOAc-Hex (1/1), Rf = 0.69, UV and PMA.



   (l) (c) (S) -4-bromo-3-hydroxybutanoic acid methyl ester Reference: the same as for the preparation of (l) (b).



   A solution purged with argon of the dibromide is treated
 EMI41.2
 (20, 80 g; 75, 4 mmol) and anhydrous NaOAc (21.0 g) in EtOAc (370 ml) and glacial HOAc (37 ml) with Pd (5 96) / C ( 1.30 g) and the black suspension is stirred under H2 (1 atmosphere) while controlling the fixation of H2. Once the fixation of H is complete (after 2.0 hours), the mixture is filtered on Celite, the filtrate is washed
 EMI41.3
 with NaHCO-, saturated and brine, it is then dried over anhydrous MUS04 and concentrated by evaporation to obtain a crude dibromoester in the form of a brown oil.

   The crude oil is combined with another fraction (starting from 36.77 g of dibromide) and they are distilled under vacuum to obtain 25.77 g (61.3%) of the bromoester mentioned in the title, desired in the form of '' a clear oil

  <Desc / Clms Page number 42>

 
 EMI42.1
 with a point a'eGuiilon cte / -XU "(J u, U mm Hg). (thin layer hromatography: EtOAc-Hex (1/1), Rf = 0.44, PMA.



  Analysis: Calculated for CeHQO .. Br: C: 30, 48; H: 4.60;
Br: 40, 56
Found: C: 29.76; H: 4.50; Br: 39, 86 (2) (S) -4-bromo-3 - [[1,1-dimethylethyl) diphenyl silyll xylbutanoic acid methyl ester
A solution of bromohydrin from part E (1) (4.0 g; 20.4 mmol), imidazole (6.94 g; 5.0 equivalents) and 4-dimethylamino pyridine (4-DMAP) is treated. (12 mg; 0.005 equivalent
 EMI42.2
 in desiccated DMF (40 ml) with t-butyldiphenylsilyl chloride (5.84 ml; 1.1 equivalents) and the homogeneous mixture is stirred overnight under argon at room temperature.

   The mixture is distributed between 5% KHIS04 and EtOAc, the organic phase is washed with H2O and brine, dried over anhydrous NaSO and concentrated by evaporation to give 9.32 g (100%) crude silyl ethe in the form of a viscous, clear oil. Thin layer chromatography: Hex-EtOAc (3/1); Rf: silylated ether = 0.75, U. V and PMA.



  (3) (S) -4-iodo-3 - [[(1,1-dimethylethyl) diphenyl silyl oxylbutanoic acid methyl ester
 EMI42.3
 A solution of the crude bromide of part E (2) (9.32 g; 201 mmol) in methyl ethyl ketone (60 ml dried on sieves of 4) is treated with sodium iodide (15, 06 g; 100, mmol; 5.0 equivalents) and the yellowed suspension is refluxed for 5.0 hours under argon. The mixture is cooled, diluted with EtOAc, filtered, the filtrate is washed with dilute NaHSO (until it becomes colorless) and brine, then dried over anhydrous Na2SO4 and concentrated by evaporation in vacuo to obtain 10.17 g of a yellow oil.

   The crude oil is purified by rapid chromatography on silica gel (600 g, eluting with a mixture of hexane and CH2 Cl2 0/1). The product fractions are combined and concentrated by evaporation to obtain 7.691 g (overall yield for the two stages of 74.29 of the iodide mentioned in the heading, desired in the form of a viscous oil

  <Desc / Clms Page number 43>

 colorless, clear. Thin layer chromatography: Hex-EtOAc (3/1), Rf of the product = 0.75, U. V. and PMA. (Note: the iodide product gives joint stains to the starting bromide).
 EMI43.1
 



  (4) (S) -4- (diethoxyphosphinyl) -3- [[(1,1-dimethylethyl) diphenylsilylloxylbutanorgue acid methyl ester
A solution of iodide (7.691 g) in triethyl phosphite (20 ml) is heated at 1550C (oil bath) for 3.5 hours under argon. The mixture is cooled and the excess phosphite (0.5 mm Hg; 75 ° C.) is distilled off under vacuum to leave a yellow oil (approximately 8.0 g). The crude oil is purified by rapid chromatography on silica gel (400 g), eluting with a mixture of hexane and acetone (4/1). The product fractions are concentrated by evaporation to obtain 3.222 g (41.1 96) of the above-mentioned phosphonate, desired in the form of a clear, colorless viscous oil.

   Thin layer chromatography: Hex-acetone (1/1), Rf = 0, 51, U. V. and PMA. In addition, 2,519 g are recovered (corrected yield of 61.1%) of iodide from the starting part (3).



  (5) (S) -3 - [[(1,1-dimethylethyl) diphenylsilyl] - oxy] -4-phosphonobutanoic acid methyl ester
A solution of the phosphonate of part (4) (9.85 g; 20.0 mmol) in CH-CL-desiccated (60 ml) is treated successively with bistrimethylsilyltrifluoroacetamide (BSTF A) (5.31 ml; 32, 0 mmol; 1.6 equivalent) and trimethylsilyl bromide (TMSBr) (6.60 ml; 50.0 mmol; 2.5 equivalent) and the clear mixture is stirred overnight under argon at room temperature. 5% KHIS04 (80 ml) is added and the mixture is extracted with EtOAc. The aqueous phase is separated with NaCl and re-extracted with EtOAc.

   The combined organic layers are washed with brine, dried over NaSO. anhydrous and concentrated by evaporation in vacuo to give the phosphonic acid mentioned in crude section in the form of a viscous oil. Layer chromatography
 EMI43.2
 thin: iPrOH-NH. OH-HO (7/2/1), Rf = 0, 30, U. V. and PMA.



  (6) Methyl ester of (S) -3- [[((1, 1-dimethylethyl) diphenylsilyl] oxy] -4- (hydroxymed thoxyphosphin y I) butanoic acid The crude phosphonic acid of the part is treated

  <Desc / Clms Page number 44>

 
 EMI44.1
 \ / \ cnvnu '). u, u) <mti \ -j / ucmo uc 10. pynumc actnc \ z. tHi / little u dried CH-OH (on 3A sieves; 1.62 ml; 40.0 mmol; 2.0 equivalent) and dicyclohexyl carbodiimide (DCC) (4.54 g; 22.0 mmol: 1 , 10 equivalent) and the resulting white slurry soc argon was stirred at room temperature overnight. The pyridin is separated in vacuo and an azeotropic distillation is then carried out with benzene (2 x 15 ml).

   The residual oil is dissolved in EtOAc, filtered, washed with 1.0 N HCl and brine, dried over NaSO. anhydrous and concentrated by evaporation in vacuo to obtain 8.272 g of ester mentioned in the gross heading in the form of an oil containing a small amount of urea dicyclohexyl
 EMI44.2
 precipitated (DCU). Thin layer chromatography: iPrOH-NH4 OH-H20 (7/2/0, Rf = 0, 60, U. V. and PMA.



  F. Methyl ester of (S) -4-ssI4'-fluoro-3, 3 ', 5-trimethyl {1 biphenyi] -2-yl] methoxyphosphmyl] -3-t-butyldiphenylsilyloxybutano! Acid ester! mono methyl ester of crude phonic phonic acid from part E (6.595 g; approximately 14.7 mmol) with dried CH2Cl2 (30 ml), it is treated with distilled trimethylsilyl diethylamine (5.60 ml; 29 , 4 mmol; 2.0 equivalents) and the mixture is stirred under argon at room temperature for one hour The mixture is concentrated by evaporation under vacuum, it is taken up in benzene (1 x 30 ml) and dried under vacuum .

   The light yellow viscous oil is dissolved in dried CH2Cl2 (30 m and DMF (dried on 4A sieves; 2 drops), the clear solution being cooled to -10OC (salt / ice bath) and treated dropwise drop by syringe with distilled oxalyl chloride (1.41 ml; 16.2 mmol; 1.1 equivalent). A violent evolution of gas is manifested and the solution becomes darker yellow. the mixture is stirred under argon at -10 ° C. for 15 minutes and the stirring is allowed to continue at room temperature for one hour. The mixture is concentrated by evaporation under vacuum, taken up in benzene (1 x 30 ml) and dried under vacuum to give crude phosphon chloridate as a yellow oil.



   To a solution of crude phosphonochloridate (approx 14.7 mmol) in dried CH2Cl2 (10 ml), dropwise

  <Desc / Clms Page number 45>

 dropwise a solution of the biphenyl alcohol of part D (2.06 g; 8.43 mmol) in dried pyridine (15 ml) and the resulting mixture is stirred at room temperature under argon for 16 hours. The mixture is concentrated by dry evaporation and the
 EMI45.1
 residue is partitioned between 5% KHS04 and EtOAc. The organic phase is washed with saturated NaHCO 4 and brine, then dried over anhydrous NaSCL and concentrated by evaporation in vacuo to obtain 8.290 g of a brown oil. The crude product is purified by rapid chromatography on silica gel (370 g), eluting with hexane-acetone (70/30).

   The product fractions are combined and concentrated by evaporation to give 3.681 g (66%) of the above-mentioned phosphonate, desired in the form of a pale yellow oil. Thin layer chromatography: hexane-acetone (3/2), Rf = 0, 59, U. V. and PMA.



  G. (5) -4-ff4'-fluoro-3, 3 ', 5-trimethyl- [1,1'- biphenyl] -2-yl] methoxyphosphinyl] -3-hydroxybutanoic acid methyl ester
A mixture of the silylated ether of part F (1.103 g; 1.66 mmol) in dried THF (20.0 ml) is treated with glacial acetic acid (380 μl; 6.64 mmol; 4.0 equivalents) and a 1.0 M tetrabutylammonium fluoride solution (4.98 ml; 4.98 mmol; 3.0 equivalents) and the light yellow solution was stirred overnight at room temperature under argon. The mixture is distributed between cold H2O and EtOAc, the organic phase is washed with saturated NaHCO and brine, dried over NaSO. anhydrous and concentrated by evaporation into a viscous yellow oil (1.174 g).



  The crude oil is purified by rapid chromatography on silica gel (47 g), eluting with CH2CL-acetone (85/15). The product fractions are concentrated by evaporation to obtain 679 mg (93.1%) of the alcohol mentioned in the heading, desired in the form of a clear viscous oil. Thin layer chromatography: hexane-acetone (1/1), Rf = 0.41, U.V. and PMA.



  H. Monolithic acid salt (S} -4-ill4'-fluoro-3, 3 ', 5-trimethyl- [1,1'- biphenyl] -2-yl] methoxy] methoxyphosphinyl] -3-hydroxybutanoic acid
A solution of the methyl ester of part G (184 mg; 0.420 mmol) in dioxane (5.0 ml) is treated with

  <Desc / Clms Page number 46>

 1.0 N LiOH (0.50 ml; 1.2 equivalent) and the mixture is stirred at room temperature under argon for 3 hours. The mixture is diluted with H2O, filtered through a 0.4 µm polycarbonate membrane and concentrated by evaporation in vacuo.
 EMI46.1
 



  The residue is dissolved in H2O (75 ml), frozen and lyophilized. The crude acid is dissolved in a minimal amount of HO and chromatographed on a 100 ml bed of CHP-20 resin, eluting with a linear gradient system of HO / CH-CN. The product fractions are concentrated by evaporation, dissolved in H2O (50 ml), filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 174 mg (89.1% relative to the weight of the hydrate) of the monolithic salt mentioned in the heading, desired in the form of a white solid. Thin layer chromatography: iPrOH-NH40H-H20 (7/2/1), Rf = 0, 58, U. V. and PMA. Analysis: Calculated for C21H25O6PFLi + 1.95 mole of H2O (P.

   M. de 465, 46): C: 54, 19; H: 6.26; F: 4.08; P: 6, 65
Found: C: 54, 19; H: 6.21; F: 4.29; P: 6.43 H1 NMR (400 HMz):
6 1.74-2.08 ppm (2H, m, -PO (OCH-) CHL-)
2.30 (3H, s, aromatic methyl)
 EMI46.2
 2.32 (3H, d, aromatic methyl in oc relative to fluorine, J HF 2.2 Hz) or "2.35-2.62 (2H, m, -CH2-CH2Li)
2.46 (3H, s, aromatic methyl)
3, 57 &
 EMI46.3
 3, 63 (3H, 2 doublets, -OP (OCH -) -, 2 diastereoisomers, JH p = 10.3 Hz) 4.28 (1H, m, -CHCH (OH) CHCOLi) 0 fi 4, 97 (2H , m, PhCHOP (OCH) R) 6, 87-7, 25 (5H, m, H aromatics)

  <Desc / Clms Page number 47>

 
EXAMPLE 2 Dilithium salt of (S) -4 - [[[4'-fluoro-3,3 ', 5-trimethyl [1,1'-biphenyl] - 2-yl] methoxy] hydroxyphosphinyl] -3-hydroxybutanoic acid salt
A solution of the diester of Example 1 (374 mg;

   0.853 mmol) in dioxane (8.0 ml) with 1.0 N LiOH (2.6 ml; 3.0 equivalents) and heated to 500C (oil bath) for 5.0 hours under argon. A white precipitate appears. The mixture is diluted with HO and filtered. The solution is extracted once with Et2O, filtered through a 0.4 µm polycarbonate membrane and concentrated in vacuo.

   The crude product is chromatographed on a CHP-20 resin (100 ml bed), eluting with a linear gradient system of H20 / CH3CN. The product fractions are concentrated by evaporation under vacuum, taken up in H2O (50 ml), filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 260 mg (67.1% relative to the weight of hydrate) of dilithium salt mentioned in the heading, desired in the form of a white solid.



  Thin layer chromatography: PrOH-NH. OH-HO (7/2/1), Rf = 0.47, U. V. and PMA.



  Analysis: Calculated for C20H2206PFLi2 + 1.77 mole of H20:
C: 52, 88; H: 5.67; F: 4.18; P: 6.82 Found: C: 52.88; H: 5.26; F: 4.24; P: 6.43
 EMI47.1
 H NMR (400 MHz, CD30D): 0 il 1) 1.69 ppm (2H, m, -OPCHCH (OH) -) 2, 26-2, 42 (2H, m, CH CO Li)
2.30 (3H, s, aromatic methyl)
2.31 (3H, d, aromatic methyl at o of F, Ihr = 1.9 Hz)
2.38 (3H, s, aromatic methyl)
 EMI47.2
 4.22 (1H, m, -CH (OH) CH2-) 0 0 4.75 (2H, m, PhCHOP-) 1 6, 86-7, 23 (5H, m, aromatic protons)

  <Desc / Clms Page number 48>

 
Example 3 Monolithic (3S) -4 - [[[4'-fluoro-3,3 ', 5-trimethyl [1,1'-biphenyl] - 2-yl] methoxy] methylphosphinyl] -3-hydroxybutanoic acid salt AT.

   (S) acid methyl ester -4 - [[chloro) methylphosphinyl] -3 - [[(1,1- dimethyiethyi) diphenylsilylloxy] butanorque
The phosphinochloridate compound mentioned in the heading is prepared as described in the first three paragraphs of part B of Example 6.



  B. (3S) -4 - [[[4'-fluoro-3, 3 ', 5-trimethyl [I, 1'- biphenyl] -2-yl] methoxy] methytiphosphinyl] -3-t acid methyl ester -butyldiphenylsilyloxy- butanole
A cooled solution (OC; ice bath) of phosphinochloridate from part A (about 2.2 mmol) and biphenyl alcohol from part C (2) of Example 1 (429 mg; 2.2 mmol; 1 , 0 equivalent) in dried CH2Cl2 (10 ml) is treated with Et3N (425 μl; 3.04 mmol; 1.4 equivalent) and 4-DMAP (27 mg; 0.22 mmol) and the solution orange is stirred at room temperature overnight under argon.

   The mixture is distributed between 5% KHIS04 and EtOAc, the organic layer is washed with brine, dried over anhydrous Na-SCL and concentrated by evaporation to obtain 1.1 g of an orange oil. The crude oil is purified by rapid chromatography on LPS-1 silica gel (44 g), eluting with EtOAc / hexane (1/1). The product fractions are combined and evaporated to obtain 298 mg (21%) of the product mentioned in the coupled section, desired in the form of a pale yellow oil. 460 mg (corrected yield of 67%) are also recovered (biphenyl alcohol) from part C (2) of starting Example 1. Thin layer chromatography: EtOAc: Hex (1/1), Rf = 0.18, U. V. and PMA.



  C. (3S) -4 - [[[4'-fluoro-3,3 ', 5-trimethyl [1,1'- biphenyl] -2-yl] methoxy] methylphosphinyl] -3-hydroxybutanoic acid methyl ester
A solution of part B silyl ether (298 mg; 0.46 mmol) in dried THF (6.0 ml) is treated with glacial HOAc (110 µl; 1.84 mmol; 4.0 equivalents) and a 1.0 M solution of tetrabutylammonium fluoride (1.43 ml; 3.1 equivalents) in THF, the resulting solution being stirred overnight under argon

  <Desc / Clms Page number 49>

 at room temperature. The mixture is distributed between cold H2O and EtOAc, the organic phase is washed with saturated NaHCO3 and brine, dried over anhydrous NaSCL and concentrated by evaporation to obtain a yellow oil (273 mg) .

   The crude oil is purified by means of rapid chromatography on silica gel LPS-1 (11 g), eluting with a mixture of hexane and acetone (3/2). The product fractions are combined and concentrated by evaporation to obtain 150 mg (80%) of the alcohol mentioned in the desired heading in the form of a viscous oil. Thin layer chromatography: Hex / acetone (1/1), Rf = 0.23, U.V. and PMA.



  D. Monolithic acid salt (3S) -4 - [[[4'-fluoro-3,3 ', 5-trimethyl [1,1'- bi phé n yl] -2-yl] methoxy] méthyiphosphinyl] - 3-hydroxybutanoic
A solution of the methyl ester of part C (150 mg; 0.367 mmol) in dioxane (3.0 ml) is treated with 1.0 N LiOH (0.44 ml; 1.2 equivalent) and stirred the resulting white suspension at room temperature under argon for 2 hours. The mixture is diluted with H2O, filtered through a 0.4 µm polycarbonate membrane and concentrated by evaporation in vacuo to a colorless glass.



   The crude product is taken up in a minimum quantity of H2O and chromatographed on HP-20 (100 ml bed), eluting
 EMI49.1
 with a linear gradient of HO / CH-CN. The product fractions are concentrated by evaporation, taken up in 1 H 20 (50 ml), filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 130 mg (79% relative to the weight of the hydrate) of lithiated salt cited in the desired heading in the form of a white solid.



  Thin layer chromatography: CH2Cl2-CH3OH-HOAc (8/1/1), Rf = 0, 52, U. V. and PMA.



  Analysis: Calculated for C21H2505FLiP + 1.73 mole HO (PM 445, 49): C: 56, 61; H: 6.44; F: 4.26; P: 6, 95
Found: C: 56, 67; H: 6.36; F: 4.31; P: 7.43 H NMR (400 MHz):
 EMI49.2
 

  <Desc / Clms Page number 50>

 
 EMI50.1
 0 il 1, 83-2, 0 (2H, m, P- (CH) CH 2, 27-2, 40 (2H, m, Lui) 2, 30 (6H, s, 2 aromatic methyls) 2, 44 ( 3H, s, aromatic methyl) 4, 26 (1H, m, -CH2CH (OH) CH2C02Li) 0 0 II (CH 4, 87 (2H, m, ArCFLOP (CHj-) 6, 90-7, 20 (5H, m, H aromatics) Example 4 (S) -4-Q [2,4-dichloro-6- [(4-fluorophenyl) methoxy] phenyl] methoxy] methoxyphosphinyl] -3-hydroxybutanoiic acid monolithic salt 2, 4-Dichloro-6- (4-fluorophenylmethoxy) benzaldehyde (Reference: J. Med Chem., 1986, 29, 167).



  A solution of 13.77 g (72.5 mmol) of 4, 6-dichloro-2-hydroxybenzaldehyde in 100 ml of DMF is stirred and 12.02 g (87 mmol) of K.-CO- are added. This mixture is heated to about 70 ° C. for 60 minutes and then 11.7 ml of 4-fluorobenzyl bromide are added. The resulting solution is stirred at 70 ° C. for 3.5 hours, then it is poured onto ice-cold HO (1.5 liters), filtered and washed with HO and recrystallized from a mixture of 'Et20 / petroleum ether. Production: 17.88 g (83%) of slightly dirty white crystals; melting point of 107-108 C.



  B. 2, 4-Dichloro-6- [(4-fluorophenyl) methoxy] benzenemethane We treat dried Et-0 (10.0 ml), cold (0 C; ice bath) with LiAlH4 (158 mg ; 4.16 mmol; 0.6 equivalent) and the gray suspension is treated dropwise with a solution of aldehyde from part A (2.06 g; 6.83 mmol) in 10 ml of dried THF. The mixture is heated to room temperature and stirred for one hour under argon. The mixture is brought to a temperature of 00C (ice bath) and cooled by successive dropwise additions of HO (160 pal), 15% NaOH (160 μl) and H2O (475 p! ). The precipitated salts are separated by filtration through NaSO. anhydrous on packed Celite (12.7 mm bed).

   The clear filtrate is concentrated by evaporation to give 2.052 g

  <Desc / Clms Page number 51>

 (98.9%) raw alcohol in the form of white crystals. Trituration with cold hexane gives 1.892 g (91.2%) of alcohol mentioned above, pure in the form of a white crystalline solid with a melting point of 72-73 C.



  Thin layer chromatography: Hex-acetone (4/1), Rf = 0.31, U.V. and PMA.
 EMI51.1
 



  Analysis: Calculated for C14Hll02Cl2F (PM of 301, 142):
C: 55.84; H: 3.68; CI: 23, 55; F: 6.31 Found: C: 55.97; H: 3.71; CI: 23.42; F: 6.30 C. Methyl ester of (S) -4-ni2,4-dichloro-6- {(4-fluorophenyl) - methoxy] phenyl] methoxy] methoxyphosphinyl] -3-t-butyldiphenylsilyloxybutanoic acid
A solution of the methyl ester of part E (6) of Example 1 (approximately 3.84 mmol) in dried CH2Cl2 (10 ml) is treated with distilled trimethylsilyidiethylamine (1.46 ml; 7.68 mmol; 2.0 equivalents) and the resulting solution was stirred at room temperature under argon for 1.0 hour.

   The mixture is concentrated by evaporation under vacuum, taken up in benzene (1 x 20 ml) and dried under vacuum to give crude silylated phosphonic acid mono methyl ester in the form of a colorless oil.



   A solution of the crude ester (approximately 3.84 mmol) in dried CH2Cl2 (10 ml) and dried DMF (one drop) is cooled to -10 ° C. (salt; ice bath) and treated dropwise with distilled oxalyl chloride (368 µl; 4.22 mmol; 1.1 equivalent).



  Gas is released from the light yellow mixture. The mixture is stirred at room temperature under argon for one hour, concentrated by evaporation under vacuum and taken up in benzene (2 x 20 ml) to obtain crude phosphonochloridate in the form of a viscous yellow oil.



   The crude phosphonochloridate (about 3.84 mmol) in dried CH2Cl2 (10 ml) at 00C (ice bath) is treated with alcohol from part B (1.15 g; 3.84 mmol; 1.0 equivalent ) monitoring
 EMI51.2
 Et3N (805 µl; 5.75 mmol; 1.5 equivalent) and 4-DMAP (47 mg; 0.384 mmol; 0.1 equivalent) and the brown mixture is stirred overnight at room temperature under argon. We distribute the mixture

  <Desc / Clms Page number 52>

 
 EMI52.1
 between 5% KHIS04 and EtOAc, the organic phase is washed with brine, dried over Na.-, SO. anhydrous and evaporated to obtain 3.197 g of a dark brown oil. The crude product is purified by rapid chromatography on silica gel (160 g), eluting with Hex-EtOAc (7/3).

   The product fractions are combined and concentrated by evaporation to obtain 594 mg (21.1%) of the phosphonate mentioned in the desired heading in the form of a yellow oil.



  In addition, 688 mg (corrected yield of 52.4%) of alcohol from the starting part B are recovered. Thin layer chromatography: Hexacetone (1/1), Rf = 0.29, U.V. and PMA.
 EMI52.2
 



  D. (S) -4-n [2,4-dichloro-6- [(4-fluorophenyl) methyloxy] phenyi] methoxy] methoxyphosphlnyl] -3-hydroxybutano acid methyl ester? than
A solution of the silyl ester of part C (578 mg; 0.788 mmol) in dried THF (8 ml) is treated with glacial HOAc (180 #l; 3.2 mmol; 4.0 equivalents) followed of a 1.0 M solution of n-Bu4NF in THF (2.36 ml; 2.36 mmol; 3.0 equivalents) and the resulting pale yellow solution is stirred overnight under argon at room temperature. We pour the mixture
 EMI52.3
 in cold HO and extracted with EtOAc (2 x). The organic phase is washed with saturated NaHCO and brine, dried over NaSO. anhydrous and evaporated to obtain 625 mg of a yellow oil.

   The crude product is purified by rapid chromatography on silica gel (31 g), eluting with a mixture of hexane and acetone (7/3). The product fractions are combined and concentrated by evaporation to give 339 mg (86.9%) of the alcohol mentioned above, desired in the form of a viscous, colorless, clear oil.



  Thin layer chromatography: Hex-acetone (1/1), Rf = 0.25, U. V. and PMA.



  E. Monolithic acid salt (S) -4 - [[[2,4-dichloro-6 - [(4-fluorophenyl) - methoxy] phenyi] methoxy] m &num; thoxyphosphinyl] -3-hydroxybutanoic acid
A solution of the phosphonate of part D (132 mg; 0.267 mmol) in dioxane (2.5 ml) is treated with 1.0 N LiOH (0.32 ml; 1.2 equivalent) and the mixture is stirred under argon at room temperature for 4.0 hours. We notice a white precipitate. The mixture is diluted with H2O, filtered and evaporated to dryness

  <Desc / Clms Page number 53>

 
 EMI53.1
 vacuum the filtrate. The residue is chromatographed on an HP-20 resin (100 ml bed), eluting with a linear gradient system of H20 / CH3CN.

   The product fractions are combined and concentrated by evaporation, taken up in H2O, filtered through a 0.4 μm polycarbonate membrane and lyophilized to give 108 mg (79% relative to the weight of hydrate) of the salt. lithium in heading, desired in the form of a white solid.



  Thin layer chromatography: CH2Cl2-CH3OH-HOAc (20/1/1), Rf = 0.41, U. V. and PMA.



  Analysis: Calculated for C. aH. gO-ClFLiP + 1, 42 mole of HO (MW of 511, 72): C: 44, 59; H: 4.10; CI: 13, 86; F: 3.71; P: 6.05 Found: C: 44.22; H: 4.09; CI: 13, 91; F: 3.72; P: 6.11 H NMR (400 MHz): 0 è 1.98-2.11 ppm (2H, m, OP (OCH3) CH2CH (OH) -) 3 z 2.26-2.45 ppm (2H, m, -CH (OH) CHLCOLi) 3, 63 & 3, 62 (3H, 2 doublets, 2 diastereoisomers, 0 tt OP (OCH3) 3) J. jr.-ll Hz) - 3 nr 4, 23 (1H, m, (-CH2CH (OH) CH2C02Li) 2-: 2 2 5, 16 (2H, s, F-PhCHO) 5, 74 (2H, d, ArCH: zOP, JHP = 6.2 Hz) --- & ttr 7, 13-7, 53 (6H, m, aromatic H) Example 5 Dilithium salt of (3S) -4-n acid [2,4-dichloro-6- [(4-fluorophenyl) methoxy] phenyl] methoxy ] hydroxyphosphinyi] -3-hydroxybutanoic A mixture of the diester of part D of Example 4 (210 mg; 0.424 mmol) in dioxane (4.0 ml) is treated with LiOH 1.0 N (1, 30 ml;

   3.0 equivalents) and the colorless solution is heated to 500C (oil bath) under argon for 3.5 hours. A white precipitate appears after 15 minutes. The mixture is diluted with H2O, filtered and the filtrate is concentrated by evaporation in vacuo. The residue is dissolved in a minimum amount of H-O and it is chromatographed on an HP-20 resin (100 ml bed), eluting.

  <Desc / Clms Page number 54>

 with a linear gradient of H2O / CH3CN. The product fractions are combined and concentrated by evaporation. The residue is taken up in HLO (50 ml), filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 175 mg (81% relative to the weight of hydrate) of dilithiated salt. in heading, desired in the form of a white solid.
 EMI54.1
 



  Thin layer chromatography: CH2Cl2-CH3OH-HOAc (8/1/1), Rf = 0.07, U. V. and PMA.



  Analysis: Calculated for C. oH .. O. ClFLi.-P + 1.70 mole HO (PM 509.62): C: 42, 42; H: 3.84; F: 3.73;
CI: 13, 91; P: 6.08 Found: C: 42.46; H: 3.90; F: 3.93; CI: 13.42; P: 5.66 H1RMN (400 MHz): 0 1 1.73-1.92 ppm (2H, m, -OP (OLi) -CHCH (OH) -)
 EMI54.2
 2.27 (1H, dd, -CH (OH) CH, CO Li, J 4 2 HH = 8.8 Hz) 2.39 (1H, dd, -CH (OH) CHCO2Li, JHH = 4.4 Hz) z 2 HH = 4.26 (1H, m, CH CH OH) CH CO Li) 2- (2 2
5.08 (2H, s, F-Ph-CHOAr)
7.03-7.53 (6H, m, aromatic H).



   Example 6 (3S) -4 - [[2,4-dichloro-6 - [(4-fluorophenyl) methoxy] - phenyl] methoxy] methylphosphinyl] -3-hydroxybutanolque acid methyl ester A. Acid methyl ester ( S) 3 - [[(1,1-dimethylethyl) diphenylsilyl] - oxy] -4- (ethoxymethylphosphinyl) tubanoic
A mixture of the iodide from part E (3) of Example 1 (4.68 g; 9.18 mmol) in methyl diethoxyphosphine (Strem Chemicals; 5.0 g; 36.7 mmol) is heated at 100 C (oil bath) for 2.5 hours and then at 1500C for an additional 3 hours under argon. A white precipitate slowly forms in the yellow solution.

   The excess phosphine is separated by vacuum distillation (0.5 mm Hg) and the crude product is purified by rapid chromatography on silica gel, eluting with a mixture of hexane-acetone (65/35). The product fractions are combined and concentrated by evaporation to give 1.590 g (38%) of the desired phosphinic ester (mixture of diastereoisomers) in the form of an oil.

  <Desc / Clms Page number 55>

 clear viscous.



  Thin layer chromatography: Hexane-acetone (3/2), Rf (2 diastereoisomers) = 0.19 and 0.22, U. V. and PMA.
 EMI55.1
 



  B. (3S) -4 {t2, 44ichloro-6 {(4-fluorophenyl) methoxy] phenyl] methoxy] methylphosphinyl] -3-t-butyidiphenyisiiyi-butanic acid methyl ester
A solution of phosphinic ester of part A (605 mg; 1.3 mmol) in dry CHCl (6.0 ml) is treated with bis (trimethylsilyl) trifluoroacetamide (BSTFA) (280 IU; 1.05 mmol; 0.8 equivalent) and trimethylsilyl bromide (TMSBr) (210 µl; 1.57 mmol; 1.2 equivalent) and the resulting solution is stirred at room temperature under argon overnight. 5% KHIS04 (15 ml) is added and the mixture is extracted with EtOAc.

   The organic phase is washed with brine, dried over NaSO. anhydrous and concentrated by vacuum evaporation to give crude phosphinic acid in the form of a colorless oil.



   A solution of the crude phosphinic acid (approximately 1.3 mmol) in dried CH2Cl2 (6.0 ml) is treated with distilled trimethylsilyl diethylamine (270 μl; 1.44 mmol; 1.1 equivalent) and the mixture is stirred. the mixture clear at room temperature under argon for 1.0 hour. The mixture is concentrated by evaporation under vacuum, taken up in benzene (1 x 15 ml) and dried under vacuum.
 EMI55.2
 



  A cooled solution (0 C; ice bath) of crude silylated phosphinic acid (about 1.3 mmol) in dried CH2Cl2 (6.0 ml) and DMF (one drop) is treated dropwise with '' a syringe with distilled oxalyl chloride (130 µl; 1.44 mmol; 1.1 equivalent). A gas evolution appears. The mixture is stirred at room temperature under argon for one hour, it is then concentrated by evaporation under vacuum, taken up in benzene (2 x 15 ml) and dried under vacuum to give crude phosphinochloridate in the form of a yellow oil .



   A cooled solution (0 C; ice bath) of phosphinochloridate (about 1.3 mmol) and alcohol from part E (6) of Example 1 (392 mg; 1.3 mmol) in dried CH2Cl2 ( 6.0 ml) is treated with Et3N (275 μl; 1.97 mmol; 1.5 equivalent) and

  <Desc / Clms Page number 56>

 4-DMAP (16 mg; 0.13 mmol; 0.1 equivalent) and the resulting yellow mixture is stirred under argon at room temperature overnight. The mixture is distributed between 5% KHIS04 and EtOAc, the organic phase is washed with brine, dried with NaSO. anhydrous and concentrated by evaporation to obtain 908 mg of crude product in the form of a dark yellow oil.

   The crude product is purified by rapid chromatography on silica gel (45 g), eluting with a mixture of hexane and EtOAc (3/2). The product fractions are combined and concentrated by evaporation to give 266 mg (28.3%) of the product mentioned in the heading, desired in the form of a colorless, clear oil. 197 mg (57%, corrected yield) of the starting alcohol are also recovered.
 EMI56.1
 



  C. (3S) -4-ff2,4-dichloro-6- [(4-fluorophenyl) methoxy] phenyl] methoxy] methylphosphinyi] -3-hydroxybutanic acid methyl ester
A solution of the silylated ester of part B (275 mg; 0.38 mmol) in dried THF (6.0 ml) is treated with glacial HOAc (90 μl; 1.53 mmol, 4.0 equivalent) and a 1.0 M THF solution of tetrabutylammonium fluoride (1.2 ml; 3.1 equivalents). The resulting solution is stirred overnight under argon at room temperature.
 EMI56.2
 ambient rature. The mixture is distributed between cold H-O and EtOAc, the organic phase is washed with saturated NaHCO and brine, dried over NaSO. anhydrous and evaporated to obtain 258 mg of a yellow oil. The crude product is purified by rapid chromatography on silica gel LPS-1 (8 g), eluting with a mixture of hexane and acetone (1/1).

   The product fractions are combined and evaporated to give 142 mg (77%) of the alcohol mentioned above, desired in the form of a clear, colorless oil.



  Thin layer chromatography: hexane-acetone (1/1), Rf = 0.20, U. V. and PMA.



  D. Monolithic acid salt (3S) -4 - [[2,4-dichloro-6 - [(4-fluorophenyl) - methoxy] phenyl] methoxy] methylphosphinyl] -3-hydroxybutanoic
A solution of Part C methyl ester (142 mg; 0.296 mmol) in dioxane (3.0 ml) is treated with 1.0 N LiOH (0.36 ml; 1.2 equivalent) and the suspension resulting white is stirred under argon at room temperature for 2.0

  <Desc / Clms Page number 57>

 hours. The mixture is diluted with H2O, filtered through a 0.4 µm polycarbonate membrane and the filtrate is concentrated by evaporation in vacuo.



   The crude product is dissolved in a minimum quantity of H2O and it is chromatographed on a bed of 100 ml of HP-20 resin, eluting with a linear gradient of H2O / CH3CN. The product fractions are combined and concentrated by evaporation. The residue is taken up in H2O, filtered through a polycarbonate membrane and lyophilized to obtain 93 mg (63% relative to the weight of hydrate) of the lithium salt mentioned in the title, desired in the form of a white solid.
 EMI57.1
 



  Thin layer chromatography: CH2Cl2-CH3OH-HOAc (8/1/1), Rf = 0, 51, U. V. and PMA.



  Analysis: Calculated for C. QH. QOClFLiP + 1.38 mole HO (MW 495.94): C: 46.01; H: 4.42; F: 3.83; CI: 14, 30; P: 6, 24 Found: C: 46, 10; H: 4.49; F: 3.82; CI: 14, 32; P: 6.43 H NMR (400 MHz): 0.531.5 ppm (3H, d, -OP (CH3) CH2-, Jp = 14.6Hz) 0 0 Il 1.87-2.10 (2H , m, -OP (OCHjCH-) 3 2, 27 (1H, dd, -CH (OH) CH CO Li JH-H = 8.4 Hz, p = 1.1 Hz) 2.38 (1H, dd, -CH (OH) CH, CO Li, z 2 1 4, 7 Hz, J-1 1 Hz) JH-H = HP = 4.29 (1H, m, -CHCH (OH) CHCOLi)
5, 16 + 5, 18 (4H, m, ArCHOP and F-PhCH2O) -)
7.11-7.52 (6H, m, aromatic)
EXAMPLE 7 Monolithic acid salt (S) -4- [[[[4'-fluoro-3,3 ', 5-trimethyl [1,1'-biphenyl- 2-yl] methyi] amino] methoxyphosphinyl] -3- hydroxybutanolque A.

   4'-fluoro-3, 3 'acid, 4'-fluoro-3,3', 5-trimethyl [1,1'-biphenyl] -2-carboxylic acid
A solution of the aldehyde of part C (2) of Example 1 (1.0 g; 4.13 mmol) in acetone (10.0 ml) at 00C

  <Desc / Clms Page number 58>

 (ice bath) is treated dropwise with 8.0 N Jones reagent (4.1 ml; excess) and the resulting brown-green suspension is stirred overnight under argon at room temperature. The excess oxidant is destroyed by the addition of isopropanol (10.0 ml) and the precipitated chromium salts are separated by filtration through a 12.7 mm Celite pad. The filtrate is evaporated, taken up in EtOAc, washed with 1.0 N HCl (2 x), NH.

   C1 saturated (2 x) and brine, then dried over anhydrous Na2SO4 and concentrated by evaporation to give 1.011 g of a green solid with a melting point of 153-154OC.



   The crude acid is purified via the dicyclohexylamine salt.



  To a solution of the crude acid in EtOAc (5.0 ml), dicyclohexylamine (DCHA) (823 111; 1.0 equivalent) is added. The solution is diluted with hexane and the precipitated crystal salt is collected
 EMI58.1
 to give 997 mg (55% relative to the aldehyde; melting point 181-183 C) of desired product in the form of a slightly dirty white crystalline DCHA salt.



   The free acid mentioned in the section is regenerated from
 EMI58.2
 DCHA salt by dividing the salt between 5% KHSOL and EtOAc. The organic phase is washed with brine, dried over anhydrous Nua2504 and concentrated by evaporation in vacuo to give 554 mg (52% relative to the aldehyde) of the above-mentioned acid, desired.



  Thin layer chromatography: CHCl-CH-OH (9/1), Rf = 0.37, U. V. and PMA.
 EMI58.3
 



  B. 4'-Fluoro-3, 3 ', 5-trimethyl [1, 1'-biphenyil-2-carboxamide A suspension of the acid of part A (554 mg; 2, 14 mmol) in dried CH2Cl2 ( 6.0 ml) and dried DMF (one drop) at 00C (ice bath) is treated drop by drop using a syringe with distilled oxalyl chloride (205 Ill; 2.35 mmol; 1.1 equivalent) and the light yellow solution is stirred under argon at room temperature for one hour. The mixture is concentrated by vacuum evaporation, taken up in benzene (2 x) and dried under vacuum to give crude acid chloride in the form of a yellow oil.

  <Desc / Clms Page number 59>

 



   A cooled mixture (0 C; ice bath) of THF (3.0 ml) and NH. Concentrated OH (2.0 ml; excess) is treated dropwise with a solution (3.0 ml) in THF of crude acid chloride and the bright orange solution is stirred at room temperature under argon for 1 0 hours. The mixture is divided between
 EMI59.1
 of H2O and EtOAc, the organic phase is washed with saturated NaHCO, HO and brine, it is then dried over anhydrous Na2SO4 and concentrated by evaporation to give 528 mg (96.1 %) of crude amide in the form of a light orange solid. Recrystallization from EtOAc-hexane provides 435 mg (79.1%) of amide mentioned in the heading, purified in the form of needles
 EMI59.2
 pale yellow in color with a melting point of 197-198 C.

   Thin layer chromatography: Et 20-acetone (1/1), Rf = 0.83, U. V. and PMA.



  G. 4'-Fluoro-3, 3 ', 5-trimethyjtl, 1'-biphenyl] -2-methanamine A cooled solution (0 C; ice bath) of dry THF (5.0 ml) is treated with LiAIH4 solid (125 mg; 3.3 mmol) and the gray suspension is treated dropwise over a period of 5 minutes with a solution of the amide of part B (424 mg; 1.65 mmol) in THF (5 , 0 ml). The resulting suspension is stirred at room temperature under argon for 2.5 hours and then heated at reflux for 45 minutes. The mixture is brought to a temperature of 00C (ice bath) and cooled by addition
 EMI59.3
 successive drip of 125 µl H2O, 123 µl 15% NaOH and 375 µl H2O. The precipitated aluminum salts are separated by filtration through NaSO. anhydrous on packed Celite.

   The clear filtrate is concentrated by evaporation under vacuum to give the crude amine in the form of a clear oil. Thin layer chromatography: Et 20-acetone (7/3), Rf = 0.60, U. V. and PMA. The amine is purified in the form of the HCl salt.



   A solution of the crude amine (about 1.65 mmol) in absolute EtOH (8.0 ml) is treated with concentrated HCl (152 µl; 1.82 mmol) and the mixture is stirred for 15 minutes at room temperature under argon. The mixture is concentrated by evaporation under vacuum into a white crystalline solid. We grind the solid

  <Desc / Clms Page number 60>

 with cold Et 2 O, it is collected by filtration and dried under vacuum to obtain 426 mg (92.4%) of amine-HCl mentioned in the heading in the form of fine white crystals.



  D. (S) -4-M4'-fluoro-3, 3 ', 5-trimethyl- [1,1'- biphenyl] -2-yl] methyl] amino] methoxyphosphinyl] -3-t methyl ester -butyldiphenyi- silyloxy butanoic
A solution of methyl ester from part E (6) of Example 1 (approximately 2.0 mmol) in dried CH2Cl2 (5.0 ml) is treated with distilled trimethylsilyl diethylamine (758 μl; 4.0 mmol; 2.0 equivalents) and the clear mixture is stirred at room temperature under argon for one hour. The mixture is concentrated by evaporation under vacuum, taken up in benzene (1 x 15 ml) and dried under vacuum.
 EMI60.1
 



  A cooled solution (0 C) of crude silylated phosphonate in CH2Cl2 (7.0 ml) and DMF (one drop) dry is treated dropwise with distilled oxalyl chloride (192 #l; 2.2 mmol; 1.1 equivalent). Gas is released from the light yellow mixture. The solution is stirred at room temperature for one hour, concentrated by evaporation in vacuo, taken up in benzene (2 x 15 ml) and dried in vacuo to obtain the crude phosphonochloridate in the form of an oil viscous, yellow.



   A cooled (0oC) solution of phosphonochloridate and biphenyl amine-HCl from part C (416 mg; 1.49 mmol) in dried CH2Cl2 (10 ml) is treated with Et3N (641 μl; 4.6 mmol; 2.3 equivalents) and 4-DMAP (24 mg; 0.2 mmol; 0.1 equivalent) and the light yellow mixture is stirred overnight at room temperature under argon. The mixture is divided between
 EMI60.2
 5% KHS04 and EtOAc, the organic phase is washed with 4 brine, dried over NaSO. anhydrous and concentrated by evaporation in vacuo to give 1.19 g of a yellow oil. The crude product is purified by rapid chromatography on silica gel (60 g), eluting with a mixture of hexane and acetone (7/3).

   The product fractions are evaporated to give 588 mg (59.5%) of a phosphonamide mentioned in the desired section in the form of a viscous, pale yellow oil. Thin layer chromatography: hexane-acetone (7/3), Rf = 0.20, U. V. and PMA.

  <Desc / Clms Page number 61>

 E.

   (S) -4-M4'-fluoro-3, 3 ', 5-trimethyl [1,1' - biphenyl) -2-yl] methyl] amino] methoxyphosphinyl] -3-hydroxybutanoic acid methyl ester
A solution of silylated ether of part D (588 mg; 0.888 mmol) in dry THF (10.0 ml) is treated with glacial HOAc (203 μl; 3.55 mmol; 4.0 equivalents) and a 1.0 M solution in THF of tetrabutylammonium fluoride (2.66 ml; 2.66 mmol; 3.0 equivalents) and the resulting solution is stirred overnight under argon at room temperature. We pour the mixture into
 EMI61.1
 cold, 0. and extracted with EtOAc. The organic phase is washed with saturated NaHCO and brine, then dried over NaSO. anhydrous and concentrated by evaporation under vacuum to obtain 600 mg of an orange oil.

   The crude product is purified by means of rapid chromatography on silica gel (36 g), eluting with a mixture of hexane and acetone (1/1). The product fractions are combined and concentrated by evaporation to give 196 mg (50.4%) of the alcohol mentioned above, desired in the form of a light orange oil. Thin layer chromatography: hexaneacetone (1/1), Rf = 0, 16, U. V. and PMA.



  F. Monolithic salt of (S) -4-nn4'-fluoro-3, 3 ', 5-trimethyl [1,1'- biphenyl] -2-yl] methyl] amino] methoxyphosphinyl] -3-hydroxybutanoic acid salt
A solution of the diester of part E (105 mg; 0.240 mmol) in dioxane (2.0 ml) is treated with 1.0 N LiOH (288 μl; 1.2 equivalent) and the white suspension is stirred under argon at room temperature for 4.0 hours. We dilute the mixture
 EMI61.2
 with H2O, it is filtered and the filtrate is concentrated by evaporation under vacuum. The residue is chromatographed on HP-20 resin (100 ml bed), eluting with a linear gradient of HO / CH-CN. The product fractions are combined and concentrated by evaporation.



  The residue is taken up in H2O (50 ml), filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 70 mg (62.7% relative to the weight of hydrate) of salt. lithium cited in the heading, desired in the form of a white solid. Thin layer chromatography: CH2Cl2-CH3OH-HOAc (20/1/1), Rf = 0.19, U.

   V. and PMA.

  <Desc / Clms Page number 62>

 Analysis: Calculated for C21H26NO5PFLi + 2.41 moles of H2O (PM of 472, 75): C: 53, 35; H: 6.57; N: 2.96; F: 4.02;
P: 6.55 Found: C: 53.35; H: 6.52; N: 2.98; F: 4, 05
P: 6.59 H1RMN (400 MHz):
 EMI62.1
 0 if 1.79-1.97 ppm (2H, m, -P (OCH3) CH2-) 3 2.26-2.44 ppm (2H, m, -CHCO-Li)
2.29 (3H, s, aromatic methyl)
2.31 (3H, d, aromatic methyl to fluorine,
JHF = 1.4 Hz)
2.47 (3H, aromatic methyl)
3.46 &
3, 50 (3H, 2 doublets, 2
 EMI62.2
 diastereoisomers, JHP = 10.5 Hz) 0 3.96 (2H, m, -PhCHNHP (OCH -) -) 4.17 (1H, m, (-CH2CH (OH) CHCCLU)
6, 84-7, 21 (5H, m, aromatic protons)
Examples 8 to 20
By following the methods as indicated hitherto and as described in the preceding Processing Examples,

   the following additional compounds can be prepared:
 EMI62.3
 

  <Desc / Clms Page number 63>

 
 EMI63.1
 
 <tb>
 <tb> Ex.
 <tb>



  No. <SEP> R <SEP> Z <SEP> n <SEP> x <SEP> Rx
 <tb> 8. <SEP> OH <SEP> # <SEP> 1 <SEP> O <SEP> H
 <tb> 0
 <tb> #
 <tb> 9. <SEP> C2HSO <SEP> # <SEP> 2 <SEP> NH <SEP> CH3
 <tb> #
 <tb> ci
 <tb> 10. <SEP> C3H7 <SEP> # <SEP> 1 <SEP> O <SEP> Li
 <tb> 11. <SEP> CH2O <SEP> # CH2-O <SEP> 2 <SEP> NH <SEP> H
 <tb> # <SEP> 2
 <tb> 3
 <tb> #
 <tb> 12. <SEP> OH <SEP> # <SEP> 2 <SEP> -O <SEP> H
 <tb> #
 <tb>
 

  <Desc / Clms Page number 64>

 
 EMI64.1
 
 <tb>
 <tb> Ex.
 <tb>



  No. <SEP> R <SEP> Z <SEP> n <SEP> X <SEP> Rx
 <tb> 14. <SEP> C5H11 <SEP> # <SEP> 1 <SEP> NH <SEP> CH3
 <tb> 15. <SEP> OK <SEP> C2H5-CH <SEP> 2 <SEP> O <SEP> OK
 <tb> 2 <SEP> 5 <SEP> d
 <tb> #
 <tb> 16. <SEP> ONa <SEP> # <SEP> 2 <SEP> O <SEP> H
 <tb> @
 <tb> 17. <SEP> CH3O <SEP> # <SEP> 1 <SEP> NH <SEP> H
 <tb> #
 <tb>
 

  <Desc / Clms Page number 65>

 
 EMI65.1
 
 <tb>
 <tb> Ex.
 <tb>



  No. <SEP> R <SEP> Z <SEP> n <SEP> X <SEP> Rx
 <tb> 18. <SEP> CH3 <SEP> # <SEP> 1 <SEP> O <SEP> H
 <tb> 19. <SEP> HO <SEP> # <SEP>. <SEP> 2 <SEP> O <SEP> Li
 <tb> 20. <SEP> CH3O <SEP> # <SEP> 1 <SEP> NH <SEP> H
 <tb>
 

  <Desc / Clms Page number 66>

 
 EMI66.1
 Example 21 (S) -4-diisopropyloxyphosphinyl) -3-t [(1,1-dimethylethyl) diphenylsilylloxyl-butano acid methyl ester; is
The iodide from part E (3) of Example 1 (45.1 mmol; 21.70 g) was stirred under high vacuum for 30 minutes. Freshly distilled triisopropyl phosphite (0.451 mole; 93.92 g; 113.37 ml) is added in one portion and the reaction mixture is stirred under argon and heated in an oil bath at 1550C for 16 , 5 hours. The mixture is then cooled to room temperature.

   The excess triisopropyl phosphite and volatile reaction products are separated by a short distillation (10 mm Hg) followed by a Kugelrohr distillation (0.50 mm Hg; 100 C; 8 hours).



  The product is further purified by rapid chromatography (column 95 mm in diameter; Merck silica gel / 15.2 cm; eluent of hexane / acetone / toluene: 6/3/1, flow rate of 5.1 cm / minute; 50 ml fractions) to obtain 17.68 g (33.96 mmol; yield of 75%) of the isopropylphosphonate listed in the form
 EMI66.2
 of a clear viscous oil. Thin layer chromatography: silica gel Rf = 0.32 (hexane / acetone / toluene: 6/3/1).



  1RMN: (270 MHz'CDCI3) 6 7.70-7.65 (m, 4H)
7, 45-7, 35 (m, 6H)
4.57-4.44 (m, 3H)
3.59 (s, 3H)
2.94 and 2.88 (2 x d, IH J = 3.7 Hz)
2.65 and 2.60 (2 x d, IH J = 7.4 Hz)
2.24-1.87 (series of m, 2H)
1.19 and 1.12 (2 x d, 12H J = 6.3 Hz)
1.01 (s, 9H)
Example 22 Salt (1/1) of dicyclohexylamine of methyl ester of (S) -4-hydroxy-methoxyphosphinyl) -3 - [[(1,1-dimethylethyl) diphenylsilyl] oxy] butanoic acid
The isopropyl phosphonate of Example 21 (30.5 mmol; 10.66 g) is stirred under argon at room temperature in 80 ml of dry CH2Cl2.

   This solution is treated dropwise (5

  <Desc / Clms Page number 67>

 minutes) with bistrimethylsilyltrifluoroacetamide (BSTFA) (32.8 mmol; 8.44 g; 8.71 ml), followed by dropwise addition (10 minutes) of trimethylsilyl bromide (TMSBr) (51.3 mmol; 7.84 g; 6.75 ml). After stirring at room temperature for 20 hours, the reaction mixture is cut with 200 ml of 5% aqueous KHIS04 and stirred intensively for 15 minutes. The aqueous layer is extracted 3 times with ethyl acetate. The organic extracts are combined, washed once with brine, dried over NaSO and concentrated in vacuo. The residue is brought twice as an azeotrope with 50 ml of toluene.

   The precipitate which forms is suspended in toluene and filtered. The filtrate is concentrated and the azeotropic formation / filtration process is repeated. The resulting filtrate is concentrated by evaporation in vacuo and then pumped under high vacuum for 5 hours.



  The resulting viscous clear oil is stirred under argon at room temperature in 50 ml of dry pyridine. This solution is treated in a single portion with dicyclohexylcarbodiimide (DCC) (22.6 mmol; 4.65 g), this operation being followed by the addition of methanol (41.0 mmol; 1.31 g; 1.67 ml). After stirring at room temperature for 20 hours, the reaction mixture is filtered through a pad of Celite in a sintered glass funnel. The celite is washed with ethyl acetate and the combined filtrates are concentrated by evaporation in vacuo. The residue is dissolved in ethyl acetate and washed twice with 5% aqueous KHSO and once with brine.

   The organic extract is dried over NaSO 4, filtered, the filtrate is concentrated and brought twice as an azeotrope with toluene, it is suspended in toluene and filtered. The resulting filtrate is again concentrated, subjected to azeotropic formation, filtered and the filtrate is concentrated by evaporation under vacuum and placed under a high vacuum for 6 hours to obtain the phosphonate monoester in the form of a clear viscous oil (10 , 2 g, yield less than 100%). Thin layer chromatography: silica gel Rf = 0.50 (nPrOH / NH. OH / HO: 7/2/1).

   The phosphonate monoester [1.21 g is left under high vacuum for 4 hours, giving 1.16 g (2.57

  <Desc / Clms Page number 68>

 mmol)] and dissolved in 10 ml of dry ethyl ether and treated dropwise with dicyclohexylamine (2.65 mmol; 0.481 g; 0.528 ml).



  The resulting homogeneous solution is left at room temperature for 7 hours, which allows significant formation of crystals to be obtained. The mixture is stored at −20 ° C. for 16 hours and is then heated to room temperature and filtered. The crystals are washed with dry, cold ethyl ether and then left under high vacuum on P 205 for 18 hours.

   The crystals are then left under a high vacuum at 450C for 4 hours, which gives 1.25 g (1.98 mmol; yield of 77%) of the dicyclohexylamine salt mentioned in the title in the form of a solid. white powder, melting point 155-156OC. Thin layer chromatography: silica gel, Rf = 0.57 [MeOH (20%) / CH2Cl2]; H NMR: (270 MHz, CDCl-). ô 7.71-7.65 (m, 4H)
7.40-7.32 (m, 6H)
4.02 (m, 1H) 3.52 (s, 3H)
3.28 and 3.22 (m, 1 H)
3.11 (d, 3H J = 11 Hz)
2.77-2.64 (m, 2H)
2.62-2.56 (m, 1H)
1.92-1.08 (m series, 22H)
1.00 (S, 9H) Spect. mass: (FAB) 632 (M & H) + IR:

   (KBr) 3466-3457 (large) 3046.3016, 2997.2937, 2858, 2836.2798, 2721, 2704.2633, 2533, 2447, 1736.1449, 1435, 1426, 1379.1243, 1231.1191, 1107 , 1074, 1061, 1051, 820 CM-1
 EMI68.1
 Analysis: Calculated for C. H.-. OPSi. C .. HN:
C: 64.63; H: 8.61; N: 2.22 Found: C: 64.51; H: 8.49; N: 2.18

  <Desc / Clms Page number 69>

 
 EMI69.1
 It should be understood that the present invention is in no way limited to the above embodiments and that many modifications can be made thereto without departing from the scope of this patent.


    

Claims (1)

 CLAIMS 1. Compound making it possible to inhibit the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase, comprising the fragment:  EMI70.1  in which X represents a group -0- or -NH-, n is equal to 1 or 2 and Z represents a hydrophobic fixation.  2. Compound according to claim 1, characterized in that the hydrophobic attachment is a lipophilic group which, when it is linked to the upper side chain of the HMG type of the molecule by the appropriate linking element (X), binds on a hydrophobic pocket of the unused enzyme by fixing the HMG CoA substrate, leading to increased power compared to the compounds in which Z = H.  3. Compound according to claim 1, corresponding to the structure:  EMI70.2  including its salts, in which R represents an OH, lower alkoxy or lower alkyl group and R represents H or a lower alkyl group, X represents a group -0- or -NH-, n is equal to 1 or 2, Z represents a hydrophobic fixation which is a group:    <Desc / Clms Page number 71>    EMI71.1    EMI71.2  where the dotted lines represent possible double bonds, R, R, RetR can be the same or different and represent a hydrogen atom, a halogen atom or a lower alkyl, haloalkyl, phenyl, substituted phenyl or  EMI71.3  YV ORY, RY representing H or an alkanoyl, benzoyl, phenyl, halophenyl, phenyl-lower alkyl, lower alkyl, cinnamyl, haloalkyl, allyl, cycloalkyl-lower alkyl, adamantyl-lower alkyl or substituted phenyl-lower alkyl: when Z represents a group :    EMI71.4     and are. 1 entlques or different and represented-  EMI71.5  c and feel H, a lower alkyl group or OH, one of the RetRs being present when the carbon to which it is attached is bonded to a double 0 6 il bond, R represents an alkyl-'C- or arylCH ,, - group ,  <Desc / Clms Page number 72>     R 6a represents a lower alkyl, hydroxy, oxo or halogen group and q is equal to 0, 1, 2 or 3, Rx representing hydrogen or a lower alkyl group in the form of free acid, in the form of a physiologically hydrolysable and acceptable ester or in the form of salt.  4. Compound according to claim 3, characterized in that X represents -O-, R represents an alkoxy group and Z represents a group:  EMI72.1   5. Compound according to claim 3, characterized in that X represents -NH-, R represents an alkoxy group, (CH2) n represents CH2 and Z represents a group:  EMI72.2  6.  Compound according to claim 4, characterized  EMI72.3  in that it consists of the methyl ester of (S) -4- [[[4'fluoro-3, 3 ', 5-trimethyl [l, l'-biphenyl] -2-yl] methoxy acid ] methoxyphosphinyl] - 3-hydroxy-butanoic acid or by its monolithic salt, by dilithiated salt of (S) -4- acid [[[4'-fluoro-3, 3 ', 5-trimethyl [l, l'- biphenyl] -2-yl] methoxy] hydroxyphosphinyl] -3-hydroxybutanoic, by the monolithic salt of (3S) -4- acid [[[[4'-fluoro-3, 3 ', 5-trimethyl [l, l -biphenyl] -2-yl] methoxy] methylphosphinyl] -3-hydroxybutanoic, by the monolithic salt of (S) -4- acid [[[2, 4-dichloro-6- [(4-fluorophenyl) methoxy] phenyl ] methoxy] methoxyphosphinyl] -3-hydroxybutanoic, by dilithium salt of (3S) -4- acid [[[2,4-dichloro-6 - [(4-fluorophenyl) methoxy] phenyl] methoxy] hydroxy- phosphinyl] -3-hydroxybutanoic,    with (3S) -4- [[2,4-dichloro-6-  <Desc / Clms Page number 73>    EMI73.1  [(4-fluorophenyl) methoxy] phenyl] methoxy] methylphosphinyl] -3-hydroxybutanol 'or its methyl ester or by the monolithic salt of (S) -4- acid [[[[[4'-fluoro-3, 3 ', 5-trimethyl [1,1'-biphenyl-2-yl] methyl] amino] - methoxyphosphinyl] -3-hydroxybutanoic.  7. hypocholesterolemic or hypolipemic composition, characterized in that it comprises a compound according to claim 3 and a pharmaceutically acceptable carrier.  8. Compound meeting the structure:  EMI73.2  wherein Ra represents a lower alkyl or lower alkoxy group, including all of its stereoisomers.  9. Compound meeting the structure:  EMI73.3  wherein R-represents a lower alkyl group or OH, including all of its stereoisomers.  10. Use of a compound according to claim 3 for the preparation of a medicament for inhibiting the biosynthesis of cholesterol.