CH676984A5 - - Google Patents

Description

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CH 676 984 A5

Description

The present invention relates to novel phosphorus-containing compounds which inhibit the activity of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase, and which are therefore useful for inhibiting the biosynthesis of cholesterol and hypocholesterolemic compositions containing such. compounds.

F.M. Singer et al., Proc. Soc. Exper. Biol. Med., 102, 370 (1959) and F.H Hulcher Arch. Biochem. Bio-phys., 146,422 (1971) show that certain mevatonate derivatives inhibit the biosynthesis of cholesterol.

Endo et al. describe in US Pat. Nos. 4,049,495, 4,137,322 and 3,983,140 an active fermentation product for the inhibition of cholesterol biosynthesis. This product is called compactin, and it has been shown by Brown et al., J. Chem. Soc. Perkin 1.1165 (1976) that it had a complex structure of mevalonolactone.

GB No. 1,586,152 describes a group of synthetic compounds of the formula

E

where E represents a direct bond, a C1-3 alkylene bridge, a vinylene bridge and the different Rs represent various substituents.

It has been found that the activity of the product of the English patent corresponds to less than 1% of the activity of compactin.

U.S. Patent No. 4,375,475 granted to Willard et al. describes hypocholesterolemic and hypolipidemic compounds having the structure where A is an H; E is a direct bond, a -CH2-, a -GH2-CH2-, a -CH2-CH2-CH2- or a -CH = CH-: Ri, R2 and R3 are each chosen from the group consisting of H, halogens, C1-4 alkyls and Cm haloalkyls, phenyl group, halogen substituted phenyls, Ct_4 alkoxy, C2-8 alkanoyloxy groups, C1-4 alkyls, or C1-4 haloakyls and OR4 where R4 is H, ai-kanoyl C2-0, benzoyl, phenyl, halophenyl, phenyl C1-3 alkyl, C1-9 alkyl, cinnamyl,

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CH 676 984 A5

C1-4 haloalkyl, allyl, C1-3 cycloalkylalkyl, C1-3 adamantyl alkyl or substituted C1-3 phenylalkyl, the substituents of which are selected from the group consisting of halogens, C1-4 aikoxy groups, Ci alkyls ^ t Cm haloalkyls; and the corresponding dihydroxy acids resulting from the opening by hydrolysis of the lactone ring, the pharmaceutically acceptable salts of these acids and the C1-3 alkyl esters substituted with C1-7 phenyl, dimethylamino or acetylamino groups of the dihydroxy acids; all of the compounds being enantiomers having a 4R configuration in the tetrahydropyran portion of the trans racemate represented by the above formula.

WO 84/02131 (PCT / EP83 / 00308) (based on US patent application No. 443,668 filed on November 22, 1982 and US patent application No. 548,850 filed on November 4, 1983) by Sandoz AG describes heterocyclic analogs of mevalonolactones and their derivatives having the structure

3 o where one of the groups R and Ro is R

R

5a and the other is C1-6 primary alkyl, C3-6 cycioalkyl or phenyl- (CH2) m-

where R4 is hydrogen, C1-4 alkyl, aikoxy C-m, (except t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyioxy,

Rs is hydrogen, C1-3 alkyl, C1-3 aikoxy, trifluoromethyl, fluoro, chloro, phenoxy or benzyioxy,

Rsa is hydrogen, C1-2 alkyl, C1-2 aikoxy, fluoro or chloro, and m is 1,2 or 3,

with the condition that Rs and Rsa must both be hydrogen when R4 is hydrogen, Rsa must be hydrogen when Rs is hydrogen, not more than one of the two groups R4 or Rs must be trifluoromethyl, not more than one of the groups R4 and Rs must not be a phenoxy and not more than one of the two groups

R4 and Rs should only be a benzyioxy,

R2 is hydrogen, C1-4 alkyl, cycioalkyl C3-6, aikoxy C1-4 (except t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyioxy,

R3 is hydrogen, C1-3 alkyl, C1-3 aikoxy trifluoromethyl, fluoro, chloro, phenoxy or benzyioxy with the condition that Rs must be hydrogen when R2 is hydrogen, not more than one groups R2 and R3 must not be a trifluoromethyl, not more than one of the groups R2 and R3 must not be a phenoxy and not more than one of the groups Ra and R3 must not be a benzyioxy,

X is a - (CH2) n- or a -CH = CH- (n = 0,1,2 or 3),

R

£ £ 6 Ì. 1 Z is -CH-CH -C-CH -COOH II 1 2 I 2 OH OH

where Re is hydrogen or C1-3 alkyl, the product being in the form of a free acid, under the

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CH 676 984 A5

in the form of a physiologically hydrolysable and physiologically acceptable ester, in the form of a lactone or a salt.

GB 2,162,179 to describes naphthyl analogs of mevalolactone useful as inhibitors of structural cholesterol biosynthesis

Hey Me where Ri is C1-3 alkyl;

Z is a group of the formula Zi or Z2:

H .GH

-W,

0

u o

(Z} (Z Ï

1 2

R7 = H, a hydrolyzable ester group or a cation.

European patent 164 698 A describes a preparation of lactones useful as antihyperolesterolemic agents by treating an amide with an organic sulphonyl halide R5 SO2X, then by removing the protective group Pr

-CHCH CHCH COOR l 21 2 7 OH OH

1

where X = halo;

Pr = carbinol protecting group;

Ri = H or CH3;

R3, R4 = H, C1-3 alkyl, or phenyl-C1-3 alkyl, the phenyl being, where appropriate substituted with a Ci-g alkyl,

an aikoxy C1-3 or a halo, -

r2 = a group of formula (A) or (B);

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CH 676 984 A5

(AT)

6 I

OR R -CH-

I

Q = R -C-

I

CH

6 3

R6 = H or OH;

R = H or GH3;

a, b, c and d = optional double bonds;

R7 = phenyl or benzyioxy, the nucleus possibly being substituted, where appropriate, by a C1-3 alkyl or a halo; R8, R9 = C1-3 alkyl or halo;

R5 = C1-3 alkyl, phenyl or mono- or di (C1-3 alkyl) phenyl.

Anderson, Paul Leroy describe in Ger. Offen. DE 3,525,256 naphthyl analogs of mega-lonolactones of the structure

Me

Me

CHCH CHCH CO R I 2 | 2 2 OH OH

Q

H OH

KS

II

Q '

where R1 is an alkyl, Z = Q, Q1; R7 = H or a hydrolyzable ester group useful as inhibitors of cholesterol biosynthesis and in the treatment of atherosclerosis,

WO 8,402,903 (based on US Patent 460,600 filed January 24, 1983 by Sandoz AG) describes mevalono-lactone analogs useful as hypolipoproteinemic agents having the structure

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f "

CH 676 984 A5

X-Z R

where the two groups together form a radical of the formula

& 1- c r - {c = c - c = c.] - or - (CH y -Il 2 4

R R

2 3

where Rs is hydrogen, C1-4 alkyl, C1-4 aikoxy (except t-butoxy) trifluoromethyl, fluoro, chloro, phenoxy or benzyioxy,

R3 is hydrogen, C1-3 alkyl, C1-3 aikoxy, trifluoromethyl, fluoro, chloro, phenoxy or benzyioxy, with the condition that no more than one of the groups R2 and R3 is trifluoromethyl, that not more than one of the groups R2 and R3 is a phenoxy and that not more than one of the groups R2 and R3 is not a benzyioxy;

Ri is hydrogen, C1-6 alkyl, fluoro, chloro or benzyioxy;

R4 is hydrogen, C1-4 alkyl, C1-4 aikoxy (except t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyioxy;

Rs is hydrogen, C1--3 alkyl, C1-3 aikoxy, trifluoromethyl, fluoro, chiaro, phenoxy or benzyioxy;

Rsa is hydrogen, C1-2 alkyl, C1-2 aikoxy, fluoro or chloro, with the condition that no more than one of the groups R4 and Rs is trifluoromethyl, that no more than one of the groups R4 and Rs is not a phenoxy, and no more than one of the groups R4 and Rs is a benzyioxy;

H (CH)

\ / 2q X is - {CH) -, C = C

2 n / ^

- (CH) H

2 q where n is equal to 0,1,2 or 3 and both q are 0 or one is 0 while the other is 1, Zest

F tf 36 z. /

-CH-CH -C-CH -COOH II

j 2 I 2 HO OH

where Re is hydrogen or C1-3 alkyl, with the general condition that -X-Z and the phenyl group carrying R4 are in position ortho to each other;

in the form of a free acid, a physiologically hydrolyzable and physiologically acceptable ester or lactone, or a physiologically acceptable salt.

European patent application 127848 (Merck & Co, Ine) describes derivatives of hydroxy-3-thia-5-ö) aryl alkanotque having structure

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CH 676 984 A5

OH ^. Q

k. ox

S (O) 1 n E J Z

where Z is:

R

n is 0.1 or 2;

E is -CH2-, -CH2-CH2-, -CH2-CH2-CH2-;

a -CH = CH-CH2- or a- CH2-GH = CH-;

Ri, R2 and R3 are for example hydrogen, chloro, bromo, fluoro, C1 alkyl, phenyl, substituted phenyl or OR7, where R7 is hydrogen, C2-8 alkanoyl, benzoyl, phenyl , substituted phenyl, C1-9 alkyl, cinnamyl, haloalkyl Cm, allyl, cycloalkyl-alkyl C1-3, adamantyl C1-3 alkyl or phenylalkyl Ct-s;

R4, R5 and R6 are hydrogen, chloro, bromo, fluoro or C1-3 alkyl; and X is for example hydrogen, a C1-3 alkyl, a cation derived from an alkali metal or an ammonium.

These compounds have antihypercholesterolemic activity resulting from their ability to inhibit 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase, as well as antifungal activity.

French patent application No. 2,596,393 filed April 1, 1986 (Sanofi SA) describes derivatives of 3-carboxy-2-hydroxypropane phosphonic acid and in particular their salts which are useful as lipid-lowering agents and which have for formula:

R O OR I 2 11/3 S 00C-CH -C-CH -P 1 2 1 2 \

OH GOLD

4

where Ri and R2 = H, lower alkyl or, where appropriate, substituted aralkyl;

R3 and R4 = H, lower alkyl or, where appropriate, substituted aryl or substituted araikyl.

These compounds are described as causing a greater decrease in the levels of cholesterol, triglycerides and phospholipids than megutol.

European patent application 142146 A (Merck & Co., Ine) describes compounds of the mevinoline type having the formula:

E 1

Z

where R1 is for example hydrogen or C1-4 alkyl; E is a —C H 2 — C H 2—, a —OH — OH— or - (CH2) i — î and

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CH 676 984 A5

Zest

1)

O

II

R

where X is -O- or -NR9 where R9 is hydrogen or C1-3 alkyl;

R7 is C2-8 alkyl; and

R8 is hydrogen or CH3;

2)

where R10, R11 and R12 are, independently, for example hydrogen, halogen or alkyl O1-4;

In accordance with the present invention, phosphorus-containing compounds are provided which inhibit the 3-hydroxy-methylglutaryl coenzyme A reductase (HMG-CoA reductase) enzyme and which are therefore useful as hypocholesterolemic agents, these compounds having the group following

0 ti

-P-CH -CH-CH -CO-

1 2 s 2

X. OH I

(CH)

1 2 n Z

where X is -O- or -NH-, n is 1 or 2 and Z is a "hydrophobic anchor".

The term hydrophobic anchor used here designates a lipophilic group which, when it is linked to the upper part of the chain of the molecule imitating the HMG group by an appropriate bond (X), binds to a hydrophobic pocket of the enzyme not used for fixing the HMG-CoA substrate, which causes an increase in activity compared with the compounds where Z = H.

In the preferred embodiments, the compounds of the invention have the formula I

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CH 676 984 A5

0 h

H J X

I R-P-CH -C-CH -CO R

1 2 12 2 X OH

I

(CH)

2 n.

Z

where R is OH, lower aikoxy or substituted or unsubstituted lower alkyl Rx is H or substituted or unsubstituted lower alkyl;

X is -O- or -NH-;

n is 1 or 2;

Z is a lipophilic group;

as well as pharmaceutically acceptable salts.

The terms "salt" and "salts" indicate the basic salts formed from mineral and organic bases. Mention may in particular be made of ammonium salts, salts of alkali metals such as lithium, sodium and potasium (which will be preferred), salts of alkaline earth metals such as calcium and magnesium salts , organic base salts such as amine salts, for example dicy-clohexylamine, benzathine, N-methyl P-glucamine salts, hydrabamine salts, and amino acid salts such as arginine, lysine, etc. Non-toxic and pharmaceutically acceptable salts will be preferred, but other salts may also be useful in the separation or purification of the product.

Examples of hydrophobic anchors in accordance with the present invention include the following nonlimiting examples

2a 1 R v R

. od * "

2b. 2

R R

q where the dotted lines represent, optional double bonds. We can mention for example

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CH 676 984 A5

5 "

R I

O

alkyl

6 a (R)

or alkyl where R1, R2, R2a and R20 may be the same or they may be different and chosen independently of one another from the group consisting of H, trifluoromethyl halogens, substituted or unsubstituted lower alkyls, haloalkyls, the group phenyl, substituted phenyls or ORY groups where RY is H, alkanoyl, benzoyl group, phenyl group, halophenyl, lower phenylalkyl, substituted or unsubstituted lower alkyl, cinnamyl group, haloalkyl, group allyl, a (substituted or unsubstituted cycioalkyl) - (substituted or unsubstituted lower alkyl), adamantyt- (substituted or unsubstituted lower alkyl) or substituted phenyl - (substituted or unsubstituted lower alkyl).

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CH 676 984 A5

When Z is

6

R 1

O

q rs and R5 'are the same or different and they are chosen from the group consisting of H, substituted or unsubstituted lower alkyls and OH;

Re is a (substituted or unsubstituted lower alkyl)

O O

he II

-C type CH -CH -C-C-3 2 / \ 7 CH R 3 '

or aryl CH2-;

R6a is substituted or unsubstituted lower alkyl, hydroxy, oxo or halogen or trifluoromethyl;

q is 0.1,2 or 3; and

R7 is substituted or unsubstituted H or lower alkyl.

Thus, the compounds of formula I include

O

11 x

IA R-P-CH -CH-CH -CO R and

'2 = 2 2

0 OH

1

(CH)

12 n Z

0

"X

IB R-P-CH -CH-CH -CO R

1 2 3 22 NH OH

I

(CH)

i 2 n Z

The term "substituted or unsubstituted lower alkyl" or "substituted or unsubstituted alkyl" used alone or in a group denotes straight and branched hydrocarbon chains containing from 1 to 12 carbon atoms in the straight chain and preferably 1 to 7 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethyl, pentyl, oc-tyl, 2,2-trimethyl, 4 pentyl, nonyl, decyl, undecyl, dodecyl, their various branched isomers, as well as the carbon chains mentioned above and bearing halogenated substituents of type F, Br, Cl, I or CF3, substituents aikoxy, aryl, (substituted alkyl or unsubstituted) -aryl, haloaryl, cycioalkyl substi11

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CH 676 984 A5

killed or unsubstituted, alkylcycioalkyl, hydroxy, alkylamino, alkanoylamino, arylcarbonylamino, nitro, cyano, thiol or alkylthio.

The term “substituted or unsubstituted cycioalkyl” used alone or in a group denotes saturated cyclic hydrocarbon groups containing from 3 to 12 carbon atoms, and preferably from 3 to 8 carbon atoms, in particular, cyclopropyl, cyclobutyl groups, cyclopentyl, cyclohexyl, cyclo-heptyl, cyclooctyl, cyclodecyl and cyclododecyl, these hydrocarbon groups can all be substituted with 1 or 2 halogens or trifluoromethyl, 1 or 2 substituted or unsubstituted lower alkyl groups, 1 or 2 lower aikoxy 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 term "aryl" or "ar" used herein denotes monocyclic and bicyclic aromatic groups containing from 6 to 12 carbon atoms in the cyclic portion such as phenyl, naphthyl, substituted phenyl or substituted naphthyl groups where the substituents on phenyl or naphthyl can be 1, 2 or 3 substituted or unsubstituted lower alkyl groups, halogens (Cl, Br or F), 1, 2 or 3 lower aikoxy groups, 1, 2 or 3 hydroxy groups, 1, 2 or 3 groups phenyl, 1, 2 or 3 alkanoyloxy groups, 1,2 or 3 benzoyloxy groups, 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 at / or 1,2 or 3 alkylthio groups, the number of substituents being preferably 3.

The terms "aralkyl", "aryl- (substituted or unsubstituted alkyl)" or "lower aryl- (substituted or unsubstituted alkyl)" used alone or in a group denote the substituted or unsubstituted alkyl groups described above having a substituent aryl, such as a benzyl.

The terms "lower aikoxy", "aikoxy", "aryloxy" or "aralkoxy" used alone or in a group denote any of the substituted or unsubstituted lower alkyl, substituted or unsubstituted alkyl, aralkyl or aryl groups bound to a d atom 'oxygen.

The terms "lower alkylthio", "alkylthio", "arylthio" or "aralkylthio" used alone or in a group denote any of the substituted or unsubstituted lower alkyl, substituted or unsubstituted alkyl, aralkyl or aryl groups bound to an atom of sulfur.

The terms "lower alkylamino", "alkylamino", "arylamino", "arylalkylamino" used alone or in a group denote any of the substituted or unsubstituted lower alkyl, substituted or unsubstituted alkyl, aryl or arylalkyl groups bound to a d atom 'nitrogen.

The term "alkanoyl" used alone or in another group denotes substituted or unsubstituted lower alkyl groups linked to a carbonyl group.

The term "halogen" or "halo" used alone or in another group denotes chlorine, bromine, fluorine, iodine, the preferred halogens being chlorine or fluorine.

The compounds having formula I which are preferred are those whose structure is as follows

II R O CH H CH

\ | 1/2 \ / 2v x

P C CO R

I ^ 2

X OH

I

CH 12 Z

where R is OH or OLÌ; Rx is Li or H;

X is O or NH; and

Zest

R

where R1 is a phenyl having a substituted or unsubstituted alkyl substituent and / or halo, trifluoromethyl

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CH 676 984 A5

where R1 is a benzyioxy which has a halo substituent;

R2 and R2a are identical and are a halogen trifluoromethyie, or a substituted or unsubstituted lower alkyl.

Z can also preferably be:

1

R

where R1 and R2 are as defined above about the compounds of formula II or Z is

6

R

'/ ViA

R

where Rs is H, CH3 or OH and R6 is

O

H

CH C r y r

3 / \ 7 CH R 3

or a substituted phenylmethyl where R7 is H or CH3.

The compounds according to formula I of the invention can be prepared according to the reaction sequence, the description of which follows.

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CH 676 984 A5

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Ccbic R -P (Oalkyl)

i 2

QSi-C (CH) III

> 3 3

It 3 3

Arbuzov reaction (Ra = substituted or unsubstituted lower alkyl

I — CH -C-CH -CÖ lower alkyl or aikoxy) 2 12 2

H

0 a H

R -P CH-CH-CH -CO alkyl

1 21 2 2 Oalkyl OSi-C (CH)

/ \ 3 3 C H C H 6 5 6 5 IV

30 1. (CH) -SiBr, CH Cl O

3 3 2 2 a'Il

IV 2. HO R - P-CH-CH-CH -CO alkyl

2 1 2 "2 2

40

OH O l

35 '

Hydrolysis of the Si-C ester (CH)

/ v 3 3 phosphoric C H C H

6 5 6 5

GO. Ra '= substituted or unsubstituted lower alkyl when Ra was substituted or unsubstituted lower alkyl

4g BV. Ra '= OH when Ra was a lower aikoxy

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CH 676 984 A5

b

VB R OH-DCC O

a 'b H

(R = OH) <r "= lower alkyl R -O — P-CH -CH-CH ~ CO

substituted or unsubstituted) _ 'f,! ,

OH O alkyl

Pyridine

I

If — C (CH)

(esterification) / \ 3 3

C H C H

6 5 6 5

VI

VI 1) (COC1); CH Cl O

2 2 2 eye

(acid Cl formation) R -P-CH -CH-CH -CO alkyl i 2 - 2 2 or 2) Z- (CH) -XH X Ô

2 n he

VA (coupling reaction) (CH) Si-C (CH)

i2 n / \ 33 (C H) N, DMAP Z CHCH

2 5 3 ^ 6565

7

(rc is a., substituted or unsubstituted lower alkyl or lower aikoxy)

vii

1) n- (C H) NF, CH COOH, O

4 9 4 3 li x

VII THF (rupture of the silyl R-P-CH -CH-CH -CO

I 2 = 2 2 ether) • X OH

I

2) OH, dioxane (hydrolysis) (CH)

l 2 n Z

As indicated by the above reaction sequence, the compounds of Formula I can be prepared by subjecting iodide A to an Arbuzov reaction of heating iodide A

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CH 676 984 A5

with a phosphonite / phosphite Ili Hl Ra-P (Oalkyl) 2

C H • 6 5 O — Si-C (CH) I 3 3

C H 6 5

I-CH -C-CH -CO alkyl 2 i 2 2 H

where Ra is substituted or unsubstituted lower alkyl or lower aikoxy. Arbuzov standard conditions allow phosphinate / phosphonate IV to be formed

0

at ii

IV R -P-CH -CH-CH -CO alkyl

1 2 z 2 2 O OSi-C (CH)

t / \ 3 3

alkyl CHCH 6 5 6 5

Phosphinate / phosphonate IV is a new compound and as such it is part of the present invention.

The phosphorate ester bond of phosphinate / phosphonate IV is then cut by treating a solution of compound IV in an inert organic solvent such as methylene chloride successively with bis (trimethylsilyl) trifluoroacetamide (BSTFA) and trimethylsilyl bromide under an inert atmosphere ( for example argon) to form the phosphinic acid VA where Ra of IV is a substituted or unsubstituted lower alkyl:

0 he

VA lower alkyl-P-CH -CH-CH -CO alkyl

1 2 1 2 2 OH O

I

If- (CH)

/ \ 3 3 C H C H 6 5 6 5

or phosphonic acid VB (where Ra of IV is a lower aikoxy):

O II

VB HO-P-CH -CH-CH -CO alkyl

12 = 22 OH O l

If— (C (CH) / \ 3 3

C H C H 6 5 6 5

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Rn

CH 676 984 A5

Compounds VA and VB are new intermediates and as such are part of the present invention.

When the phosphonic acid VB is obtained, it is esterified by treatment in dry pyridine with an alcohol

VC RbOH (where Rb is substituted or unsubstituted lower alkyl)

and dicyclohexyl carbodiimide, the reaction mixture obtained being stirred under an inert atmosphere (for example argon), which makes it possible to obtain the monoalkyl ester of phosphonic acid VI:

O

b H

VI R -O-P-CH -CH-CH -CO alkyl

12 = 22 OH O

t

Si-C (CH)

/ \ 3 3 C H C H 6 5 6 5

Ester VI or phosphonic acid VA is then dissolved in an inert organic solvent such as methylene chloride, benzene or tetrahydrofuran (THF) and treated with trimethylsilyldi-ethylamine. The mixture is stirred under an inert atmosphere (argon). The mixture is evaporated and dissolved in methylene chloride (or another suitable inert organic solvent). The solution obtained is then cooled to a temperature between about 0 ° C and 25 ° C, treated with oxalyl chloride and evaporated. The crude phosphonochloridate is thus obtained. The phosphonochloridate is dissolved in an inert organic solvent such as methylene chloride, benzene, pyridine or THF; the solution is then cooled to a temperature between about -20 ° C and about 0 ° C and treated with

B Z- (CHz) n-XH

using a VI or VA: B molar ratio in the range of from about 0.5: 1 to about 3: 1 and preferably from about 1: 1 to about 2.1, then with triethylamine and catalytic 4-dimethylamino pyridine (DMAP) to form compound VII

0

VS "

VII R -P — CH —CH-CH -CO alkyl

1 2 = 2 2

X O

he

(CH) Si-C {CH) _

"2 n / \ 3 S Z C H C H 6 5 5 5

where Rc is substituted or unsubstituted lower alkyl or lower aikoxy.

The silyl ester bond of compound VII is cut by treating VII in an inert organic solvent such as tetrahydrofuran with glacial acetic acid and tetrabutylammonium fluoride to form the ester VIII

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CH 676 984 A5

0

He X

VIII R -P-CH -CH-CH -CO R

12 = 22 X OH

1

(CH)

| 2 il Z

x

(R = alkyl)

Ester VIII can then be hydrolyzed to obtain the alkali metal salt or the corresponding acid where R * is an alkali metal or an H. For this, treatment is carried out with a strong base, such as lithium hydroxide in the presence dioxane, tetrahydrofuran or another inert organic solvent. This treatment is carried out under an inert atmosphere (for example under argon) at 25 ° C. and using a base-ester Vili molar ratio in the range from approximately 1: 1 to 1.1: 1. The corresponding alkali metal salt is thus obtained

0 he

VIIIA R-P-CH -CH-CH -CO alkali metal

1 2 z 2 2 X OH

i

(CH}

i 2 n Z

where R is substituted or unsubstituted lower alkyl or lower aikoxy.

Compound VIIIA can then be treated with a strong acid such as HCl to form the corresponding V11IB acid.

0 ii

VIIIB R-P-CH -CH-CH -CO H

1 2 = 2 2 X OH

l

(CH)

I 2 n Z

Ester VIII where R is a lower aikoxy can be transformed to obtain the corresponding alkali metal di-salt. For this, the ester VIII is treated with a strong base at 50-60 ° C using a Vili baserester molar ratio in the range from 2: 1 to about 4: 1, which allows VIlIC to be obtained.

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CH 676 984 A5

0 11

VIIIC alkali metal-O-P-CH -CH-CH -CO Taëtal alkaline

12 = 22 X OH

1

(CH)

l 2 n z

The alkali metal di-salt VIIIC can be converted to the corresponding acid (R = OH) by treatment with a strong acid such as HCl. We then obtain

0M

VIIID HO-P-CH -CH-CH -CO H

1 2 = 2 2 X OH

!

(CH)

1 2 n Z

Starting iodide A can be prepared from bromide C

OH 1

C Br-CH -CH-CH CO alkyl

2 2 2

[prepared according to the methods described in Tetrahedron Lett. 26, 2951 (1985)]. For this, it is dissolved in dimethylformamide (DMF), imidazole and 4-dimethylamino pyridine. The solution obtained is treated with t-butyldiphenylsilyl chloride under an inert atmosphere (for example under argon) to form the silyl ether D

C H C H 6 SS / 6 5 D OSi-C (CH)

• l 3-3

Br-CH -CH-CH CO alkyl 2 2 2

A solution of silyl ether D in an inert organic solvent such as methyl ethyl ketone or DMF is treated with sodium iodide under an inert atmosphere (for example under argon) to form iodide A.

The starting compound B

B Z- (CH2) n-XH

can be prepared as described below. The synthesis depends on the nature of Z and that of X.

So the compound of formula B where Z is

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CH 676 984 A5

N R

and X is an O, and which therefore has the following structure

Bl can be prepared by treating aldehyde E

R2a "€ $ 7 (CVn ~ ° H

the

E R ~ \ ()) (CH> -CHO

r2 2 n-1

with a reducing agent such as lithium aluminum hydride or sodium borohydride. The compounds of formula B where Z is and X is N, i.e. the compounds of the structure

2a

(CH) ~ NH 2 2 n 2

can be prepared by oxidizing aldehyde E in acetonic solution with, for example, Jones' reagent to form acid F

y R

(CH) -COOH 2 2 n-1

The latter is treated in suspension in methylene chloride with chloride oxatyie to form the corresponding acid chloride. This is dissolved in an inert organic solvent such as tetrahydrofuran and treated with a mixture of concentrated ammonium hydroxide in tetrahydrofuran to form an amide having the structure

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CH 676 984 AS

O 11

(CH) -C-NH 2 2 n-1 2

To obtain amine B2, amide G is treated with a reducing agent such as lithium aluminum hydride.

The starting compounds of the formula B where Z is

51

R

q and X is an O or -NH-, that is to say compounds having the structure

(CH) -XH 2 n

6a alkyl

(R)

q where X is an O are described by C. H. Heathcock et al. in J. Org. Chem. 50.1190 (1985). The compounds of formula H 'where X is NH can be prepared by the reductive amination of

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CH 676 984 A5

alkyl

(prepared according to C. H. Heathcock et al., see above) by treating J with ammonium acetate and sodium cyanoborohydride in the presence of an alcohol such as methanol as solvent.

The starting compounds of formula B where Z is and X is O, i.e. the compounds of the structure

M

(CH} -OH 2 2 n are described in WO 8402-903-A and GB 2 162 179A both deposited by Sandoz. The starting compounds of formula B, where Z is and X is NH, that is ie compounds of the structure

NOT

R R

can be prepared by the reductive amination of aldehyde O

) -NH 2 n 2

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CH 676 984 A5

2a

1

O

R

R

2b

R R

(CH) CHO 2 2 n-1

by treating O with ammonium acetate and sodium cyanoborohydride in the presence of an alcohol such as methanol as solvent.

The compounds of the invention can be prepared as racemic mixtures and then separated to obtain the preferred isomer (S isomer). However, the compounds of the invention can be prepared directly in the form of the S isomer as described in the concrete examples below.

The compounds of the invention are inhibitors of 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase and are therefore useful for the inhibition of cholesterol biosynthesis, as demonstrated by the tests which follow .

1) Rat HMG-CoA reductase

The activity of rat hepatic HMG-CoA reductase is measured using a modification of the method described by Edwards (Edwards, P.A. et al., J. Lipid Res. 20:40, 1979). The rat liver microsomes are used as the source of the enzyme, and the enzyme activity is determined by measuring the conversion of "C-HMG-CoA (substrate) to mevalonic C".

at. Preparation of microsomes

The liver is removed from 2-4 Sprague Dawley rats fed cholestyramine and killed by decapitation. The livers are homogenized in a phosphate buffer A (potassium phosphate 0.04 M, pH 7.2; KCl, 0.05 M; sucrose, 0.1 M; EDTA, 0.03 M: aprotinin 500 Kl units / ml ). The homogenized livers are centrifuged at 16,000 g for 15 minutes at 4 ° C. The supernatant is separated and centrifuged under the same conditions a second time. The supernatant from the second centrifugation at 16,000 g is centrifuged at 100,000 g for 70 minutes at 4 ° C. The microsome pellet is resuspended in a minimum of buffer A (3-5 ml per liver) and homogenized in a glass / glass homogenizer. Dithiothreitol (10 mM) is then added. The solution is divided into aliquots which are quickly frozen in an acetone / dry ice mixture and stored at -80 ° C. The specific activity of the first microsomal preparation was 0.68 nmol of mevalonic acid / mg protein / minute.

b. Enzyme tests

The reductase is measured in 0.25 ml of a solution containing the following components, given in final concentration:

The mixtures are incubated at 37 ° C. Under the conditions described, the enzymatic activity increases linearly up to 300 μg of microsomal protein in the reaction mixture. The reaction is also linear as a function of the incubation time up to 30 minutes. The standard incubation time chosen for the study of drugs is 20 minutes, which corresponds to a 12-15% conversion of the HMG-CoA substrate to mevalonic acid. The substrate [DL-] HMG-CoA is used at a rate of 100 nM, which corresponds to twice the concentration necessary to saturate the enzyme in the con-

Potassium phosphate pH 7.0 KCl

Sucrose EDTA

Dithiothreitol NaCl

Dimethyl sulfoxide microsomal proteins

14n_rm / n ru; we.

14C- [DL] HMG-CoA (0.05 nCi 30-60 mCi / mmole) 100 | xl

NADPH (nicotinamide adenine dinucleotide phosphate) 2.7 nM

0.04 M 0.05 M 0.10 M 0.03 M 0.01 M 3.5 mM 1%

50-200 ng

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CH 676 984 A5

described editions. NADPH is used in excess at 2.7 times the concentration necessary to have the maximum enzymatic reaction speed.

Standardized tests to assess inhibitors are performed according to the following procedure. The microsomal enzyme is incubated in the presence of NADPH at 37 ° C for 15 minutes. DMSO is added with or without the test compound, and the incubation is continued for 15 minutes at 37 ° C. The enzymatic reaction begins with the addition of the substrate 14C-HMG-CoA. After 20 minutes of incubation at 37 ° C., the reaction is stopped by the addition of 25 μl of 33% KOH. 3H-mevalonic acid (0.05 fiCi) is added and the mixture is left to stand at room temperature for 30 minutes. Fifty µl of 5N HCl is added at this time to cause lactonization of mevalonic acid. Bromophenol blue is added to serve as a pH indicator to monitor the decrease in pH. Lactonization takes place in 30 minutes at room temperature. The reaction mixture is centrifuged for 15 minutes at 2800 rpm. The supernatants are placed on 2 grams of anionic resin AG 1-X8 (Biorad, formate form) and the whole is poured into glass columns of 0.7 cm in diameter. Elution is carried out with 2.0 ml of water. The first 0.5 ml are discarded, and the following 1.5 ml are collected and their 3H and 14C radioactivities are measured in 10 ml of Opti-fluor scintillating liquid. The results are given in nmoles of mevalonic acid produced in 20 minutes and corrected to 100% recovery of tritium. The effect of the drug is expressed by I50, which is the concentration of the drug causing a 50% reduction in enzyme activity. The results are obtained from several response curves as a function of the dose. We give the 95% confidence interval.

The transformation of the drugs in the form of lactones into corresponding sodium salts is carried out by dissolving the lactone in DMSO, adding an excess of sodium hydroxide corresponding to 10 times the molar quantity of lactone present and allowing to react at room temperature for 15 minutes. The mixture is then partially neutralized (pH 7.5-8.0) with 1N HCl and added to the reaction mixture containing the enzyme.

2) Synthesis of cholesterol in freshly prepared rat hepatocytes.

Compounds showing an inhibiting effect on HMG-CoA reductase were evaluated for their ability to inhibit the incorporation of i-C-acetate into cholesterol from suspensions of freshly prepared rat hepatocytes. The method used is that initially described by Capuzzi et al. (Cappuzzi D. M. and Margolis S., Lîpids, 6: 602.1971).

at. Preparation of rat hepatocytes

Sprague Dawley rats (180-220 g) are anesthetized with Nembutol (50 mg / kg). The abdomen is open and your first branch of the portal vein is closed. Heparin (100-200 units) is injected directly into the inferior vena cava. A ligature is placed on the distal section of the portal vein and a cannula is placed between this ligature and the first branch of the vein. The liver is then perfused at a speed of 20 ml / minute with oxygenated buffer A and preheated to 37 ° C (HBSS without calcium or magnesium containing 0.5 mM EDTA), after having sectioned the vena cava to allow the evacuation of the effluent The liver is then perfused with 200 ml of preheated buffer B (HBSS containing 0.5% bacterial collagenase). After the infusion with buffer B, the liver is excised and decapsulated in 60 ml of Waymouth medium, thus allowing the free cells to disperse in the solution. The hepatocytes are collected by centrifugation for 3 minutes at low speed (50 g) and at room temperature. The hepatocyte pellet is washed with Waymouth medium. A count and a viability test are then carried out by excluding the trypane blue. The hepatocytes of these cell suspensions usually have a viability of 70-90%.

b. Incorporation of ^ C-acetate into cholesterol

The hepatocytes are suspended in a proportion of 5 × 10 6 cells per 2.0 ml of incubation medium (Ml) [0.02 M Tris-HCl (pH 7.4), 0.1 M KCl, sodium citrate 3.3 mM, nicotinamide 6.7 mM, 0.23 mM NADP, glucose-6-phosphate 1.7 nMj.

The test compounds are normally dissolved in DMSO or DMSO: HaO (1: 3), then added to MI. The final concentration of DMSO in MI is less than or equal to 1.0% and has no significant effect on the synthesis of cholesterol.

Incubation begins when "^ C-acetate (58 mCi / mmoIes, 2 nCi / ml) is added and the cell suspension (2.0 ml) is placed in wells for tissue culture. 35 mm, incubation is carried out at 37 ° C. and lasts 2.0 hours After incubation, the cell suspensions are transferred to glass centrifuge tubes and centrifuged for 3 minutes at 50 g at room temperature. The cell pellets are resuspended, lysed in 1.0 ml H2O and placed in ice.

The lipids are extracted as described by Blight E. G. and W. J. Dyer, Can. J. Biochem. and PhysioL, 37: 911, 1959. The lower organic phase is separated and dried under a stream of nitrogen and the residue is suspended in 100 μl of chloroform: methanol (2: 1). The entire sample is deposited on silica gel plates for LK6D thin layer chromatography. Development is done with a

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CH 676 984 A5

hexane mixture: ethyl ether: acetic acid (75: 25: 1). The radioactivity of the plates is determined using the BioScan automatic scanning reading system. The total radioactivity of the cholesterol spot (Rf 0.28) is expressed relative to the total radioactivity of the lipid extract. The cholesterol spots in the reference cultures contained 800-1000 cpm and this radioactivity corresponded to 9-20% of the total radioactivity of the lipid extract. These results are comparable to those of Capuzzi et al., Which indicates that cholesterol contains 9% of the radioactivity of the lipid extract.

The effect of the drugs (% inhibition of cholesterol synthesis) is determined by comparing the% of radioactivity incorporated into cholesterol in the reference cultures and in the cultures treated with the drug. Response curves are developed by combining the results of two or more trials. The results are expressed by I50 accompanied by the value of the 95% confidence interval.

3) Synthesis of cholesterol in human skin fibroblasts

The inhibitory effect of cholesterol biosynthesis in liver tissue constitutes a possible selection criterion. In addition to examining the inhibitory effect on the biosynthesis of cholesterol in hepatocytes, the compounds of the invention are tested for their inhibitory action on the biosynthesis of cholesterol in cultured fibroblasts.

at. Culture of human skin fibroblasts

Human skin fibroblasts are cultured on Eagle's minimum essential medium (ME) containing 10% fetal calf serum. In each test, the cell culture monolayers are dispersed by ultrasound. The cells are then counted and placed in 35 mm wells for tissue culture (5 x 105 cells / 2.0 ml). The cells are incubated for 18 hours at 37 ° C. in a chamber under humid air containing 5% of CO2. The induction of cholesterol biosynthesis enzymes is done by removing the serum-enriched medium, washing the cell monolayers and adding 1.0 ml of ME containing 1.0% bovine serum albumin without free fatty acids. The incubation is then extended for 24 hours.

b. Incorporation of 14C-acetate into cholesterol

The induced fibroplast cells are washed with MEME100 (Earie's minimum essential medium). The test compounds are dissolved in DMSO or DMSO: ME (1: 3) and added to the cell culture (the final concentration of DMSO in the cell culture medium is equal to or less than 1.0%). After preincubation for 30 minutes at 37 ° C under humidified air containing 5% CO2 and presence of the drug, [1-14C] sodium acetate (2.0 (j.Ci / ml, 58 mCi / mmol) is added The incubation is then continued for 4 hours, the culture medium is removed after the incubation and the cell monolayer (200 ng of cellular proteins per culture) is scraped into 1.0 ml of water. extracts from the cell suspension lysed with chloroform: methanol, as described with regard to hepatocyte suspensions. The organic phase is dried under nitrogen and the residue is suspended in 100 μl of chloroform: methanol ( 2: 1). The entire sample is applied to a silica gel plate for thin layer chromatography (LK6D). The plates are analyzed as described for hepatocytes.

The inhibition of cholesterol synthesis is determined by comparing the percentage of radioactivity found in the cholesterol spot of the treated cultures and the reference cultures. Response curves are obtained by combining the results of two or more trials. The results are expressed by I50 accompanied by the value of the 95% confidence interval.

Another aspect of the present invention relates to pharmaceutical compositions comprising at least 1 compound of formula I in combination with a pharmaceutical carrier or a diluent. The pharmaceutical composition can be formulated using conventional liquid or solid vehicles of a type suitable for the desired mode of administration. Administration can be oral, for example in the form of tablets, capsules, granules or powders, or it can also be parenteral in the form of injections, such galenical forms containing from 1 to 2000 mg of active compound per unit dose used in treatment. The amounts administered depend on the unit dose, symptoms, age and weight of the patient.

The compounds of formula I can be administered in the same way as known compounds such as lovastatin used for the inhibition of the biosynthesis of cholesterol in mammals, and in particular in humans, dogs, cats, etc. Thus, the compounds of the invention can be administered at a rate of 4 to 2000 mg in a single daily dose or in 1-4 doses distributed over the day, with a preference for an administration of 4 to 200 mg per day divided into several doses from 1 to 100 mg, or better still administration in 2-4 times a day of doses ranging from 0.5 to 50 mg; one can also use dosage forms with a persistent effect.

The following Practical Examples constitute preferred forms of the present invention. Unless otherwise indicated, all temperatures are expressed in degrees centigrade. The Chro-

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CH 676 984 A5

Rapid matography was carried out either on Merck 60 silica gel or on Whatmann LPS-1 silica gel. Reverse phase chromatography was performed on CHP-20 MCI resin gel supplied by Mitsubishi Ltd.

Example 1

Mono-lithium salt of (S) acid - [[[fluoro-4'trimethyl-3,3 ', 5 [biphenyl-1,1'] yl-2] methoxy] methoxyphosphinyl] -4 hydroxy-3 butyric acid

A. N- (2,4-dimethyl benzylidene) benzeneamine.

(US Patent Ref. 4,375,475 granted to Merck, p. 39).

A solution of freshly distilled 2,4-dimethyl benzaldehyde (Aldrich, 6.97 ml, 50 mmol) and distilled aniline (Aldrich, 4.56 ml, 50 mmol) in dry toluene (80.0 ml) was heated at reflux for three hours under argon in a flask fitted with a Dean-Stark device. The mixture was cooled and evaporated in vacuo until a yellow oil was obtained. The crude oil was purified by Kugelrohr distillation (0.5 mm Hg, 160-180 ° C) to obtain 8.172 g (78.1%) of the desired benzeneimine in the form of a light yellow oil which crystallizes at rest to provide a low melting point solid.

Hexane-acetone thin layer chromatography (4: 1) Rf = 0.67 and 0.77 (geometric isomers), UV and h.

(U.S. Patent Ref. 4,375,475 to Merck, p. 39).

Benzenimine A (6.0 g, 28.7 mmol) in glacial acetic acid (144 ml) was treated with palladium (II) acetate (6.44 g, 28.7 mmol) and the homogeneous, clear and red-colored solution was heated at reflux under argon for one hour. The cloudy mixture obtained was filtered hot through a 1/2 inch compact bed of cellite and collected in 900 ml of H2O. The precipitated orange solids were collected by filtration and dried for 16 hours under vacuum at 65 ° C in the presence of P2O5. There was thus obtained 10.627 g (85.5%) of the palladium complex sought which was in the form of an orange solid product with a melting point of 194 ° -196 ° (the melting point of an analytical sample recrystallized given by the literature is 203-205 ° C).

C. Fluoro-4'trimethyl-3,3 ') 5 [biphenyl-1,1'] - 2 carboxaldehyde

(1) Bromo [4-fluoro-3-methylphenyl] magnesium

(US Patent Ref. 4,375,475 granted to Merck, pp. 37 and 38).

Grignard's reagent C (1) was prepared by adding 5-bromo-2 fluoro-toluene (22.5 g, 60.9 mmol, Fairfield Chemical Co.) dropwise and at a speed sufficient to maintain the reaction medium at reflux at magnesium turns (1.35 g, 55.4 mmol, 8.0 eq) in anhydrous EfeO (70.0 ml). The reaction was started using an ultrasound device. When the addition of the bromide was complete, the mixture was further stirred for 1 hour at room temperature, then heated to reflux for 15 minutes and finally allowed to cool to room temperature.

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CH 676 984 AS

(2) FIuoro-4'trimethyl-3,3 ', 5 [biphenyl-1,1'] - 2 carboxaldehyde

In a second container, the complex of palladium B (3.0 g, 6.92 mmol) and latriphenylphosphine (14.52 g, 55.4 mmol, 8) was mixed for 30 minutes at room temperature and under argon. 0 eq) dissolved in dry benzene (100 ml). Compound C (1) freshly prepared and filtered through glass wool was added to this solution all at once by means of a burette, and the mixture was stirred for 1.5 hours at room temperature and under argon. 6.0N hydrochloric acid (35 ml) was then added, and the mixture was stirred for an additional hour at room temperature, then filtered through a compact bed (1/2 inch) of cellite. The filtrate was extracted with Et20 (250 ml), the extract washed with brine (2 times 100 ml), dried over anhydrous MgSCH and evaporated in vacuo to provide 13.35 g of a viscous orange oil which crystallizes at rest. The crude orange solid was purified by flash chromatography on silica gel (700 g) with elution with hexane, followed by hexane-ET20 (95: 5). The product-containing fractions were evaporated to yield 1.507 g (89.9%) of the desired aldehyde which appeared as a light yellow solid having a melting point of 72-75 ° C. (The melting point according to the literature is 73-74 ° C).

Hexane-EtgO thin layer chromatography (95: 5) Rf = 0.40, UV and APM.

D. Fluoro-4'trimethyl-3,3 ', 5 [1-biphenyl, l'] - 2 methanol

A cold solution (0 ° C, ice bath) of dry EfeO (15.0 ml) was treated with LÌAIH4 (259 mg, 6.82 mmol, 0.55 eq). Aldehyde C (3.0 g, 12.4 mmol) dissolved in dry Et2O (15 ml) was then added dropwise over 15 minutes to the gray suspension obtained. The mixture was stirred at room temperature under argon for 30 minutes, then it was cooled to 0 ° C and the reaction blocked by the dropwise addition of 260 µl of H2O, followed by 260 (xi of NaOH 15 % and finally 780 or H2O. The suspension was diluted with EtOAc, filtered through a compact bed of 1/4 inch thick cellite and the colorless filtrate was evaporated in vacuo to give 2.99 g ( 98.8%) of a white solid. Trituration of this crude solid with cold hexane and drying under vacuum made it possible to collect 2.467 g (81.6%) of the desired alcohol in the form of a white solid having a melting point of 102-103 ° C.

Hexane-EtOAc thin layer chromatography (9: 1) Rf = 0.24, UV and APM.

E. (S) - [[(1,1-dimethyl) diphenylsilyl] oxy] -3 (hydroxymethoxyphosphilyl) -4 butyric acid methyl ester

(1) (S) -bromo-4-hydroxy-3-butyric acid methyl ester

(1) (a) Hydrated calcium salt of [R- (R *, R *)] - 2,3,4-trihydroxy butyric acid (Ref. Carbohydrate Research 72, p. 301-304 (1979)) .

Calcium carbonate (50 g) was added to a solution of D-isoascorbic acid (44.0 g, 250 mmol) in water (625 ml). The suspension was cooled to 0 ° C in ice and treated with successive portions of 30% H2O2 (100 ml). The mixture was then stirred at 30-40 ° C (oil bath) for 30 minutes. “Darco” active carbon (10 g) was added and the black suspension was heated on a boiling water bath until the production of O2 was stopped. The suspension was then filtered through cellite, evaporated in vacuo (bath temperature, 40 ° C). The residue was taken up in water (50 ml), heated on a boiling water bath and CH3OH was added until the solution became cloudy. The rubbery precipitate was collected by filtration and air dried to give 30.836 g (75.2%) of the desired calcium salt as a powdery solid.

Thin layer chromatography iPrOH-NI-UOH-hfeO (7: 2: 1) Rf = 0.19, APM).

(1) (b) Methyl ester of [S- (R *, S *) j 2,4-dibromo-3-hydroxy-butyric acid (Ref. Bock, K. et al., Acta Scandinavia (B) 37 p 341-344 (1983))

The calcium salt (1) (a) (30 g) was dissolved in acetic acid containing 30-32% HBr (210 ml) and stirred at room temperature for 24 hours. Methanol (990 ml) was then added to the brown solution which was then stirred overnight. The mixture was evaporated, and an orange oil was obtained. This was taken up in GH3OH (75 ml), heated under reflux for 2.0 hours and evaporated. The residue was taken up in a mixture of EtOAc (100 ml) and H2O. The organic phase was washed twice with water and a saline solution and then dried over anhydrous NaaSO * and evaporated, which gave 22.83 g (90.5%) of crude dibromide present under the form of a light orange oil.

EtOAc-hexane thin layer chromatography (1: 1) Rf = 0.69, UV and APM.

(1) (c) Methyl ester of (S) - 4-bromo-3-hydroxy butyric acid (Ref. See product (1) (b)).

A solution of EtOAc (370 ml) and anhydrous HOAc (37 ml) purged with argon containing dibro-

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CH 676 984 A5

ripe (20.80 g, 75.4 mmol) and anhydrous NaOc (21.0 g) was treated with 5% Pd / C (1.30 g) and the black suspension was stirred under hydrogen (1 atmosphere ). The absorption of hydrogen was followed. After 2.0 hours, the absorption of hydrogen was complete. The mixture was then filtered through cellite, and the filtrate was washed with a saturated NaHCO 3 solution and with a saline solution, then dried over anhydrous MgSC> 4 and finally evaporated to obtain the crude dibromoester in the form of an oil. Brown. The crude oil was combined with another preparation (made from 36.77 g of dibromide) and distilled under vacuum, to obtain 25.77 g (61.3%) of the desired bromoester in the form of an oil Clear having a boiling point of 79-80 ° C (1.0 mm Hg).

EtOAc-hexane thin layer chromatography (1: 1) Rf = 0.44, APM.

Composition calculated for C5H9O3 Br: C, 30.48; H, 4.60; Br, 40.56.

Composition found: C, 29.76; H, 4.50; Br, 39.86.

(2) Methyl ester of (S) -bromo-4 [[(1,1-dimethyl) diphenylsilyl] oxy} -4-butyric acid

A solution of bromohydrin E (1) (4.0 g, 20.4 mmol), imidazole (6.94 g, 5.0 eq) and dimethyl-4-amino-pyridine (4-DMAP) (12 mg, 0.005 eq) in dry DMF (40 ml) was treated with t-butyldi-phenylethyl chloride (5.84 ml, 1.1 eq) and the homogeneous mixture was mixed overnight under argon at temperature ambient. The mixture was taken up in a mixture of EtOAc and a 5% solution of KHSÖ4. The organic phase was washed with water and brine, dried over anhydrous Na2SO4 and evaporated to give 9.32 g (100%) of the crude silyl ether as a clear, viscous oil.

Hexane-EtOAc thin layer chromatography (3: 1), Rf = 0.75, UV and APM.

(3) Methyl ester of (S) -iodo-4 [[(1,1-dimethylethyl) diphenylsilyljoxyj-3 butyric acid.

A solution of crude bromide E (2) (9.32 g, 201 mmol) in methyl ethyl ketone (60 ml> dried on a 4 A molecular sieve) was treated with sodium iodide (15.06 g, 100.5 mmol, 5.0 eq) and the yellow suspension was heated under reflux for 5 hours under argon. The mixture was cooled, diluted with EtOAc, filtered, and the filtrate washed with dilute NaHSOs (until discoloration) and a saline solution, and finally dried over anhydrous Na2SO4 and evaporated in vacuo to give 10.17 g of 'a yellow oil. This crude oil was purified by flash chromatography on silica gel (600 g) eluting with hexane-C ^ Clz (3: 1). The product fractions were combined and evaporated to give 7.691 g (overall yield for the two stages: 74.2%) of the desired iodide in the form of a colorless and viscous oil,

Hexane-EtOAc thin layer chromatography (3: 1) Rf = 0.75, UV and APM. (Note: the starting iodide and bromide migrate in the same way).

(4) Methyl ester of (S) - (diethoxyphosphinyl) -4 [[(1,1-dimethyl) diphenylsilyl] oxy] -3 butyric acid.

A solution of iodide (7.691 g) in triethyl phosphite (20 ml) was heated at 155 ° C (oil bath) for 3.5 hours under argon. The mixture was cooled and the excess phosphite removed by vacuum distillation (0.5 mm Hg, 75 ° C) to leave a yellow oil (about 8.0 g). The crude oil was purified by flash chromatography on silica gel (400 g), elution being carried out with hexane-acetone (4: 1). The product-containing fractions were evaporated to give 3.222 g (41.1%) of the desired phos-phonate in the form of a colorless, viscous oil.

Hexane-acetone thin layer chromatography (1: 1) Rf = 0.51, UV and APM. In addition, 2.519 g of the starting iodide (3) were recovered, which gives a corrected yield of 61.1%.

(5) (S) - [[(1,1-dimethyl-ethyl) diphenylsilyi] oxy] -3-phosphono-4-butyric acid methyl ester.

A solution of phosphonate (4) (9.85 g, 20.0 mmol) in dry CH2Cl2 (60 ml) was successively treated with bistrimethylsilyltrifluoroacetamide (BSTFA) (5.31 ml, 32.0 mmol, 1.6 eq) and tri-methylsilyl bromide (TMSBr) (6.60 ml, 50.0 mmol, 2.5 eq) and the clear mixture was stirred overnight under argon at room temperature. A 5% KHSO4 solution (80 ml) was then added and the mixture was extracted with EtOAc. The aqueous phase was saturated with NaCl and reextracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and evaporated in vacuo to give the desired phosphonic acid as a viscous oil.

Thin layer chromatography iPr0H-NH40H-H20 (7: 2: 1) Rf = 0.30, UV and APM,

(6) Methyl ester of (S) - [[(1,1-dimethylethyl) diphenyisily!] Oxy] -3 (hydroxymethoxyphosphinyl) -4 butyric acid

The crude phosphonic acid (5) (about 20.0 mmol) was treated in solution in dry pyridine

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CH 676 984 A5

(25 ml) with dried CH3OH (3 Å molecular sieve, 1.62 ml, 40.0 mmol, 2.0 eq) and dicyclohexyl carbodiimide (DCC) (4.54 g, 22.0 mmol, 1 , 10 eq). The white suspension thus obtained was stirred under argon at room temperature overnight. The pyridine was removed in vacuo and then azeotropic distillation was carried out with benzene (2 x 15 ml). The residual oil was dissolved in EtOAc and this solution was filtered and washed with 1.0 N HCl and a saline solution, dried over anhydrous NaaSO * and finally evaporated in vacuo to give 8.272 g of the ester sought. This crude product was an oil containing a small amount of precipitated dicyclohexylurea (DCU).

Thin layer chromatography iPr0H-NH40H-0H2 (7: 2: 1) Rf = 0.60, UV and APM.

F. (S) acid methyl ester - [[[fluoro-4'trimethyl-3,3 ', - 5 [bipheny 1-1,1'] yI-2] methoxyphosphinyi] -4 t-butyldiphenylsilyloxy-3 butyric

The monomethyl ester of crude phosphonic acid (E) (6.595 g, about 14.7 mmol) was dissolved in dry CH2Cl2 (30 ml), treated with trimethylsilyldiethylamine (5.60 ml, 29.4 mmol , 2.0 eq) and stirred under argon at room temperature for 1 hour. The mixture was evaporated in vacuo, removed with benzene (1 time 30 ml) and dried in vacuo. The light yellow viscous oil was dissolved in dry CH2Cl2 (30 ml) and DMF (dried over 4 Å molecular sieve, 2 drops). The clear solution was cooled to -10 ° C (salt / ice) and added to it dropwise with a syringe of distilled oxalyl chloride (1.41 ml, 16.2 mmol, 1.1 eq). A significant emission of gas occurred and the solution took on a dark yellow color. The mixture was stirred for 15 minutes under argon at -10 ° C and then for 1 hour at room temperature. The mixture was evaporated in vacuo, removed with benzene (1 time 30 ml) and dried in vacuo to give the crude phosphonochloridate as a yellow oil.

To a solution of crude phosphonochloridate (about 14.7 mmol) in dry CH2Cl2 (10 ml), a solution of biphenyl alcohol D (2.06 g, 8.43 mmol) in dry pyridine was added dropwise (15 ml). The resulting mixture was stirred at room temperature under argon for 16 hours. The mixture was evaporated to dryness and the residue taken up in a mixture of EtOAc and a 5% solution of KHSO4. The organic phase was washed with a saturated NaHCOg solution and with a salt solution, then dried over Na2SO4 and evaporated in vacuo to give 8.290 g of a brown oil. The crude product was purified by flash chromatography on silica gel (370 g) eluting with hexane-acetone (70:30). The product-containing fractions were combined and evaporated to give 3.681 g (66%) of the desired phosphonate in the form of a pale yellow oil.

Hexane-acetone thin layer chromatography (3: 2) Rf = 0.59, UV and AMP.

G. Methyl ester of (S) acid - [[[fluoro-4 'trimethyl-3,3', 5 [biphenyl-1,1 '] yl-2] [methoxyphosphinyl] -4 hydroxy-3 butyric

A mixture of silyl ether F (1.103 g, 1.66 mmol) and dry THF (20.0 ml) was treated with glacial acetic acid (380 µl, 6.64 mmol, 4.0 eq) and with a 1.0 M solution of tetrabutylammonium fluoride (4.98 ml, 4.98 mmol, 3.0 eq). The light yellow solution obtained was stirred overnight at room temperature and under argon, then taken up in a cold mixture of EtOAc and H2O. Then, the organic phase was washed with a saturated NaHCOa solution and with a salt solution, dried over anhydrous NagSO4, then evaporated to obtain a yellow viscous oil (1.174 g). The crude oil was purified by flash chromatography on silica gel (47 g) and elution with CH2Cl2-acetone (85:15). The product-containing fractions were evaporated to obtain 679 mg (93.1%) of the desired alcohol in the form of a viscous oil.

Hexane-acetone thin layer chromatography (1: 1) Rf = 0.41, UV and APM.

H. Acid mono-lithium salt (S) - [[[fluoro-4 'trimethyl-3,3', 5 [biphenyl-

I.1 '] yI-2] methoxy] methoxyphosphinyl] -4 3-hydroxybutyric

A solution of methyl ester G (184 mg, 0.420 mmol) in dioxane (5.0 ml) was treated with 1.0 N LiOH (0.50 ml, 1.2 eq) and the mixture was stirred at room temperature under argon for 3 hours.

The mixture was diluted with H2O, filtered through a 0.4 µm polycarbonate membrane and evaporated in vacuo. The residue was dissolved in H2O (75 ml), frozen and lyophilized. The crude acid was dissolved in a minimum quantity of water and chromatographed on 100 ml of a bed of CHP-20 resin, elution being carried out with a linear gradient system H2O / CH3CN. The fractions containing the product were evaporated, and the latter was dissolved in H2O (50 ml), and the solution was filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 174 mg (89.1% based on the weight of the hydrate) of the mono-lithium salt sought.

Thin layer chromatography iPr0H-NH40H-H20 (7: 2: 1) Rf = 0.58, UV and APM.

Composition calculated for C21H25O6PFLÌ + 1.95 moles of H2O (PM, 465.46): C, 54.19; H, 6.26; F, 40.8; P, 6.65,

Composition found: C, 54.19; H, 6.21; F, 4.29; P, 6.43.

H1 NMR (400 MHz:

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CH 676 984 A5

S 1.74-2.08 ppm (2H, m, -PO (OCH3) CH2-)

2.30 (3H, s, methyl aromatic)

2.32 (3H, d, aromatic methyl has a with respect to a fluorine, Jhf 2.2 Hz)

2.35-2.62 (2H, m, -CH2C02Li)

2.46 (3H, s, methyl aromatic)

3.57 & 3.63 (3H, 2 doublets, -OP (OCH3) - 2 diastereomers, Jh-p = 10.3 Hz)

4.28 (1H, m, -CH2CH (0H) CH2C02Li)

O II

4.97 (2H, m, PhCH OP (OCH) R)

2 3

6.87-7.2 (5H, m, Aromatic H)

Example 2

(S) - [[[fluoro-4 'trimethyl- 3,3', 5 [biphenyl-1,1 '] yl-2] methoxy] hydroxyphosphinyi] -4 hydroxy-3-butyric acid di-lithium salt

A solution of the diester of Example 1 (374 mg, 0.853 mmol) in dioxane (8.0 ml) was treated with 1.0 N LiOH (2.6 ml, 3.0 eq) and heated to 50 ° C (oil bath) for 5.0 hours under argon. A white precipitate appeared. The mixture was diluted with H2O and filtered. The aqueous solution was extracted once with Et 2 O, filtered through a 0.4 µm polycarbonate membrane and concentrated in vacuo. The crude product was chromatographed on a 100 ml bed of CHP-20 resin, eluting with a linear gradient system H2O / CH3CN. The product-containing fractions were evaporated in vacuo, the residue was taken up in H2O (50 ml), and the solution was filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 260 mg (67.1% on the basis of the hydrate weight) of the desired lithium lithium salt in the form of a white solid.

PrOH-NH40H-H2O thin layer chromatography (7: 2: 1) Rf = 0.47, UV and APM.

Composition calculated for C2oH2206PFLi + 1.77 moles of water: C, 52.88: H, 5.67; F, 4.18; P, 6.82. Composition found: G, 52.88; H, 5.26; F, 4.24; P, 6.43.

H1 NMR (400 MHZ, CD3OD):

Where he

51.69 ppm {2H, m, -OPCH CH (OH) -)

2

2.26-2.42 (2H, m, CH2C02Li)

2.30 (3H, s, methyl aromatic)

2.31 (3H, d, aromatic methyl in a with respect to F, Jhf = 1.9 Hz)

2.38 (3H, s, methyl aromatic)

4.22 (1H, m, -CH (OH) CH2-)

O

U

4.75 (2H, m, PhCH OP-)

2 I

6.86-7.23 (5H, m; aromatic protons)

Example 3

Mono-lithium salts of (3S) acid - [[[fluoro-4 'trimethyl-3,3', 5 [biphenyl-1,1 '] yl-2] methoxy] [methylphosphinyl] -4 hydroxy-3 butyric

A. (S) - [(chloro) methylphosphinyl] -4 [[(dimethyl-1,1 ethyl) diphenyIsilyl] -oxy] -3 butyric acid methyl ester

This phosphinochloridate was prepared as described in Example 6, part B, first three paragraphs.

B. Methyl ester of (3S) acid - [[[fluoro-4 'trimethyl-3,3', 5 [biphenyl-1,1'Jyl-2] methûxy] methylphosphinyl] -4 t-butyldiphenylsilyloxy-3-butyric

A cooled solution (0 ° C, ice bath) of phosphinochloridate A (about 2.2 mmol) and of biph-

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CH 676 984 A5

alkyl alcohol C (2) of Example 1 (429 mg, 2.2 mmol, 1.0 eq) in dry CH2Cl2 (10 ml) was treated with Et3N (425 µl, 3.04 mmol, 1, 4 eq) and 4-DMAP (27 mg, 0.22 mmol) and the orange solution obtained was stirred at room temperature under argon overnight. The mixture was taken up in a mixture of EtOAc and a 5% solution of KHSO. The organic layer was washed with brine, dried over anhydrous Na2SO4 and evaporated to obtain 1.1 g of an orange oil. The crude oil was purified by flash chromatography on LPS-1 silica gel (44 g) eluting with PEtOAc: hexane (1: 1). The product containing fractions were combined and evaporated to give 298 mg (21%) of the desired product as a pale yellow oil. 460 mg of the biphenyl alcohol C (2) from the starting Example 1 were also collected, which gives a corrected yield of 67%. EtOAc thin layer chromatography: hexane (1: 1) Rf = 0.18, UV and APM,

C. (3S) - [[[4-trimethyl-trimethyl-3,3 ', 5 [biphenyl-1,1'-2-jyl-2] methoxy] methylphosphinyl] -4 hydroxy-3-butyric acid methyl ester

A solution of silyl ether B (298 mg, 0.46 mmol) in dry THF (6.0 ml) was treated with glacial HOAc (110 µl, 1.84 mmol, 4.0 eq) and with a solution 1.0 M tetrabutylammonium fluoride (1.43 ml, 3.1 eq). The resulting solution was stirred overnight under argon at room temperature, then taken up in a cold mixture of H2O and EtOAc. The organic phase was then washed with a saturated NaHCOa solution and with a salt solution, then dried over anhydrous Na2SO4 and evaporated to obtain a yellow oil (273 mg). The crude oil was purified by rapid chromatography on silica gel LPS-1 (11 g), elution being carried out with hexane-acetone (3: 2). The fractions containing the product were combined and evaporated to obtain 150 mg (80%) of the desired alcohol in the form of a viscous oil.

Hexane thin layer chromatography: acetone (1: 1) Rf = 0.23, UV and APM.

D. Acid mono-lithium salt (3S) - [[[4-fluoro-3,3 'trimethyl, 5 [1,1' biphenyl] yl-2] [methoxy] methylphosphinyl] -4 hydroxy -3 butyric

A solution of methyl ester C (150 mg, 0.367 mmol) in dioxane (3.0 ml) was treated with 1.0 N LiOH (0.44 ml, 1.2 eq) and the resulting white suspension stirred at room temperature under argon for 2 hours. The mixture was diluted with H2O, filtered through a 0.4 µm polycarbonate membrane and evaporated in vacuo to obtain a colorless solid product.

The crude product was taken up in a minimum quantity of H2O and chromatographed on a bed of HP-20 (100 ml), eluting with a linear gradient system H2O / CH3CN. The product-containing fractions were evaporated, the residue was taken up in H2O (50 ml), and the solution was filtered through a 0.4 μm polycarbonate membrane and lyophilized to obtain 130 mg (79% based on the hydrate weight) of the desired lithium salt in the form of a white solid.

Thin layer chromatography CH2CI2-CH3OH-HOAC (8: 1: 1) Rf = 0.52, UV and APM.

Composition calculated for C21H25O5FLÎP + 1.73 moles of H2O (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 H1 NMR (400 MHz):

o

1.49 ppm (3H, d, -OP C CHs) -, jh-p = 14.7 Hz)

O

1.83-2.00 ppm (2H, m, p - (CH3) CH «->

2.27-2.40 ppm (2H, m, CH2CO2LÌ)

2.30 ppm (6H, s, 2 aromatic methyls)

2.44 ppm (3H, s, 1 aromatic methyl)

4.26 ppm (1 H, m, -CH2ÇH (0H) CH2C02U)

O

he

4.87ppm (2H, m, ArCHoOP <CH,) ->

6.90-7.20 ppm (5H, m, aromatic hydrogens)

Example 4

Mono-lithium salt of (S) acid - [[[2,4-dichloro [(4-fluoro-phenyl) -6-methoxyphenyljmethoxyjmethoxy phosphinyl] -4 hydroxy-butyric

A. 2,4-Dichloro (4-fluoro-phenylmethoxy) -6 benzaldehyde (Ref .: J. Med. Chem. 1986,29,167)

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CH 676 984 A5

To a solution of 13.77 g (72.5 mmol) of 4,6-dichloro-2-hydroxy-benzaldehyde in 100 ml of DMF with stirring, 12.02 g (87 mmol) of K2CO3 are added. The mixture is heated to about 70 ° C for 60 minutes, then 11.7 ml of 4-fluoro benzyl bromide is added. The mixture is stirred at 70 ° C. for 3.5 hours, it is poured into an ice / water mixture (1.5 l), it is filtered, then it is washed with H2O and recrystallized from EfeO / petroleum ether. Yield: 17.88 g (83%) of whitish crystals, melting point 1Q7-108 ° C.

B. 2,4-Dichloro [4-fluoro-phenyl] methoxy] -6 benzene methanol.

Dry Et20 (10.0 ml) and cold (0 ° C, ice bath) were treated with L1AIH4 (158 mg, 4.16 mmol, 0.6 eq) and the gray suspension was treated dropwise with a solution of aldehyde A (2.06 g, 6.93 mmol) in 10 ml of dry THF. The mixture was warmed to room temperature and stirred for one hour under argon. The mixture was cooled to 0 ° C (ice bath) and the reaction was blocked by the sequential addition of H2O (160 (il) 15% NaOH (160 ni) and H2O (475 ni The precipitated salts were separated by filtration over anhydrous Na2SO4 placed on compact cellite (1/4 inch bed). The clear filtrate was evaporated to give 2.052 g (98.9%) of crude alcohol. in the form of white crystals. Trituration with cold hexane gave 1.892 g (91.2%) of the desired pure alcohol in the form of a crystalline solid having a melting point of 72- 73 ° C.

Hexane-acetone thin layer chromatography (4: 1) Rf = 0.31, UV and APM.

Composition calculated for C14H11O2CI2F (PM, 301.142): C, 55.84; H, 3.68; Cl, 23.55; F, 6.31,

Composition found: C, 55.97; H, 3.71; Cl, 23.42; F, 6.30.

C. (S) - [[[2,4-dichloro-4-phenyl) methoxy-6-pheny]] methoxy] methoxyphosphinyl] -4 t-butyldiphenylsilyloxy-3-butyric acid methyl ester

A solution of the methyl ester E (6) of Example 1 (approximately 3.84 mmol) in dry CH2Cl2 (10 ml) was treated with distilled trimethylsilyl diethylamine (1.46 ml, 7.68 mmol, 2.0 eq) and the solution obtained was stirred at room temperature under argon for 1.0 hour. The mixture was evaporated in vacuo, removed with benzene (1 time 20 ml) and dried in vacuo to give the crude silylated phosphonic acid mono-methyl ester as a colorless oil.

A solution of the crude ester (about 3.84 mmol) in dry CH2Cl2 (10 ml) and dry DMF (1 drop) was cooled to -10 ° C (salt / ice) and treated dropwise with distilled chloride oxalyl (368 fil) 4.22 mmol, 1.1 eq). The yellow mixture produced a gas emission. The mixture was stirred at room temperature under argon for one hour, evaporated in vacuo, removed with benzene (2 times 20 ml) to give the crude phosphonochloridate as a viscous yellow oil.

The crude phosphonochloridate (about 3.84 mmol) was treated in dry ChfeCfe (10 ml) at 0 ° C (ice bath) with alcohol B (1.15 g, 3.84 mmol, 1.0 eq ), then with Et3N (805 (il, 5.76 mmol, 1.5 eq) and 4-DMAP (47 mg, 0.384 mmol, 0.1 eq). The brown mixture was stirred overnight at at room temperature under argon, then it was taken up in a mixture of EtOAc and a 5% KHSO4 solution. The organic phase was washed with brine, dried with anhydrous Na2SO4 and evaporated to give 3.197 g of dark brown oil. The crude product was purified by flash chromatography on silica gel (160 g) eluting with hexane-EtOAc (7: 3). The product containing fractions were combined and evaporated to give 594 mg (21.1%) of the desired phosphonate as a yellow oil. In addition, 688 mg of starting alcohol B was collected, which gives a corrected yield of 52.4%. .

Hexane-acetone thin layer chromatography (1: 1) Rf = 0.29, UV and APM.

D. Methyl ester of (S) - [[[2,4-dichloro [(4-fluoro-phenyl)] -6-methoxylphenyllmethoxylmethoxy-phosphinyl] -4 hydroxy-3-butyric acid

A solution of silyl ester C (578 mg, 0.788 mmol) in dry THF (8 ml) was treated with glacial HOAc (180 nl, 3.2 mmol, 4.0 eq), then with a solution of 1.0 M of n-Bu4NF in THF (2.36 ml, 2.36 mmol, 3.0 eq). The pale yellow solution obtained was stirred overnight under argon at room temperature. The mixture was poured onto cold H2O and extracted with EtOAc (2 times). The organic phase was washed with saturated NaHCO3 solution and with brine, then dried with anhydrous Na2SO4 and evaporated to give 625 mg of a yellow oil. The crude product was purified by flash chromatography on silica gel (31 g), eluting with phexane-acetone (7: 3). The product-containing fractions were combined and evaporated to give 339 mg (86.9%) of the desired alcohol in the form of a colorless, viscous oil.

Hexane-acetone thin layer chromatography (1: 1) Rf = 0.25, UV and APM.

E. Mono-lithium salt of (S) - [[[2,4-dichloro [(4-fluorophenyl) methoxy] -6 phenyl] methoxy] methoxyphosphinyl] -4 hydroxy-3-butyric acid.

A solution of phosphonate D (132 mg, 0.267 mmol) in dioxane (2.5 ml) was treated with

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CH 676 984 A5

1.0 N LiOH (0.32 ml, 1.2 eq) and the mixture was stirred under argon at room temperature for 4.0 hours. A white precipitate was observed. The mixture was diluted with H2O, filtered and the filtrate evaporated to dryness under vacuum. The residue was chromatographed on HP-20 resin (100 ml bed), elution being carried out with a linear gradient system H2O / CH3CN. The fractions containing the product were combined and evaporated, then taken up in H 2 O and the solution obtained filtered through a 0.4 μm polycarbonate membrane to give 108 mg (79% based on the weight of the hydrate) of the desired lithium salt in the form of a white solid.

Thin layer chromatography CH2CI2-CH30H-H0Ac (20: 1:!) Rf = 0.41, UV and APM.

Composition calculated for C19H18O7CI2FIÌ2P + 1.41 moles of H2O (PM, 511.72): C, 44.59: H, 4.10; Cl, 13.86; F, 3.71; P, 6.05.

Composition found: C, 44.22; H, 4.09, Cl, 13.91; F, 3.72: P, 6.11. H1 NMR (400 MHz);

. Where he

1.98-2.11 ppm {2H, m, OP (OCH) CH CH (OH) -

3 2

2.26-2.45 ppm (2H, m -CH (0H) CH2C02Li)

3.63 & 3.62 (3H, 2 doublets, 2 diastereomers)

O

11

OP {OCH) CH -, JHP-11 HZ)

3 2

4.23 (1H, m, (-CH2CH (0H) CH2C02Li)

5.16 (2H, s, F-PhCHaO)

5.24 (2H, d, ArCH2OP, J = 6.2 Hz)

7.13-7.53 (6H, m, Aromatic H)

Example 5

(3S) - [[[2,4-dichloro-4-phenyl) methoxy] -6 phenyl] methoxy] hydroxyphosphinyl] -4 hydroxy-3-butyric acid di-lithium salt

A mixture of the diester D of Example 4 (210 mg, 0.424 mmol) and dioxane (4.0 ml) was treated with 1.0 N LiOH (1.30 ml, 3.0 eq) and the colorless solution was heated to 50 ° C (oil bath) under argon for 3.5 hours. A white precipitate appeared after 15 minutes. The mixture was diluted with H2O, filtered and the filtrate evaporated in vacuo. The residue was dissolved in a minimum quantity of H2O and chromatographed on HP-20 resin (100 ml bed), elution being carried out with a linear gradient system H2O / CH3CN. The product-containing fractions were combined and evaporated. The residue was taken up in H2O (50 ml), and the solution was filtered through a 0.4 nm polycarbonate membrane and lyophilized to give 175 mg (yield of 81% based on the weight of the hydrate) of the desired lithium salt in the form of a white solid.

Thin layer chromatography CH2CI2-CH3OH-HOAC (8: 1: 1) Rf = 0.07 UV and APM.

Composition calculated for C18H18O7CI2FU2P + 1.70 moles of water (MW, 509.62): C, 42.42; H, 3.84; F, 3.73; Cl, 13.91; P, 6.08.

Composition found: C, 42.46; H, 3.90; F, 3.93; Cl, 13.42; P, 5.66.

NMR "" H (400 MHz):

O 11

S 1.73-1.92 ppm (2H, m, —OP (OLi) -CH CH (OH) -

2

2.27 (1 H, dd, -CH (0H) CH2C02Lî, Jhh = 8.8 Hz)

2.39 (1 H, dd, -CH (0H) CH2C02Li, Jhh = 4.4 Hz)

4.26 (1H, m, CH2CH (0H) CH2C02Li)

5.08 (2H, s, F-Ph-CH2OAr)

7.03-7.53 (6H, m, Aromatic H)

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CH 676 984 A5

Example 6

(3S) - [[2,4-Dichloro-4-phenyl) methoxy-6-phenyl] methoxy] methoxyphosphinyl] -4 hydroxy-3-butyric acid methyl ester

A. (S) - [[(1,1-dimethylethyl) diphenylsilyl] oxy] -3 (ethoxymethylphosphinyl) -4-butyric acid methyl ester

A mixture of iodide E (3) of Example 1 (4.68 g, 9.18 mmol) and methyldiethoxyphosphine (Strem Chemicals, 5.0 g, 36.7 mmol) was heated under argon to 100 ° C (oil bath) for 2.5 hours, then at 15 ° C for three more hours. A white precipitate gradually formed in the yellow solution. The excess phosphine was removed by vacuum distillation (0.5 mm Hg) and the crude product was purified by rapid chromatography on silica gel, elution being carried out with hexane-acetone (65 : 35). The product containing fractions were combined and evaporated to give 1.590 g (38%) of the desired phosphinic ester (a mixture of diastereomers) in the form of a clear viscous oil.

Hexane-acetone (3: 2) thin layer chromatography Rf (2 diastereomers) “0.19 and 0.22, UV and AMP.

B. Methyl ester of (3S) - [[2,4-dichloro [(4-fluoro phenyl) methoxy] -6 phenyljmethoxyjmethylphosphinyl-4 t-butyldiphenylsilyl-3 butyric

A solution of phosphinic ester A (605 mg, 1.3 mmol) in dry CH2Cl2 (6.0 ml) was treated with bis (trimethylsUyl) trifluoroacetamide (BSTFA) (280 (il. 1.05 mmol, 0.8 eq) and trimethylsilyl bromide (TMSBr) (210 μl, 1.57 mmol, 1.2 eq). The solution obtained was stirred at room temperature under argon overnight. A 5% solution of KHSO4 (15 ml) was added, and the mixture was extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous [\ Ia2SO4 and evaporated in vacuo to give the crude phosphinic acid under the form of a colorless oil.

A solution of the crude phosphinic acid (about 1.3 mmol) in dry CH2Cl2 (6.0 ml) was treated with distilled latrimethylsilyl diethylamine (270 µl, 1.44 mmol, 1.1 eq) and the mixture Clear was stirred at room temperature under argon for 1.0 hour. The mixture was evaporated in vacuo, removed with benzene (1 time 15 ml) and dried in vacuo.

A cooled solution (0 ° C, ice bath) of crude silylated phosphinic acid (about 1.3 mmol) in dry CH2Cl2 (6.0 ml) and DMF (1 drop) was treated dropwise with using a syringe with distilled chloride oxalyl (130 ni. 1.44 mmol, 1.1 eq). A gas emission has occurred. The mixture was stirred at room temperature under argon for one hour, then evaporated in vacuo, stripped with benzene (2 x 15 ml) and dried in vacuo to give the crude phosphinochloridate as a yellow oil.

A cooled solution (0 ° C, ice bath) of the phosphinochloridate (approximately 1.3 mmol) and alcohol E (6) of Example 1 (392 mg, 1.3 mmol) in dry CH2Cl2 (6 , 0 ml) was treated with EfeN (275 nl> 1.97 mmol, 1.5 eq) and 4-DMAP (16 mg, 0.13 mmol, 0.1 eq). The yellow mixture obtained was stirred under argon at room temperature overnight and was then taken up in a mixture of a 5% solution of KHSO4 and EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SO4 and evaporated to give 908 mg of crude product as a dark yellow oil. The crude product was purified by flash chromatography on silica gel (45 g), elution being carried out with hexane-EtOAc (3: 2). The product-containing fractions were combined and evaporated to give 266 mg (28.3%) of the desired product as a colorless oil. 197 mg of the starting alcohol were also recovered (which gives a corrected yield of 57%).

C. Methyl ester of (3S) - [[2,4-dichloro [(4-fluoro-pheny!) 6-methoxyj-phenyljmethoxy] methylphosphinyl] -4 hydroxy-3-butyric

A solution of silyl ester B (275 mg, 0.38 mmol) in dry THF (6.0 ml) was treated with glacial AcOH (90 µl, "1.53 mmol, 4.0 eq) and a solution 1.0 M tetrabutylammonium fluoride in THF (1.2 ml, 3.1 eq) The solution obtained was stirred overnight under argon at room temperature, and it was then taken up in a cold mixture of H2O and EtOAc. The organic phase was washed with a saturated NaHCOa solution and with a saline solution, dried over anhydrous Na2SC> 4 and evaporated to give 258 mg of a yellow oil. The crude product was purified by Rapid chromatography on silica gel LPS-1 (8 g), elution being carried out with hexane-acetone (1 l) The fractions containing the product were combined and evaporated to give 142 mg (77%) sought after alcohol in the form of a colorless oil.

Hexane-acetone thin layer chromatography (1: 1) Rf = 0.20, UV and APM.

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D. Mono-lithium salt of (3S) - [[2,4-dichloro [(4-fluorophenyl) methoxy-6-phenyl] methoxy] methylphosphinyl] -4-hydroxy-3-butyric acid

A solution of methyl ester C (142 mg, 0.296 mmol) in dioxane (3.0 ml) was treated with a solution of 1.0 N LiOH (0.36 ml, 1.2 eq) and the suspension The resulting white was stirred under argon at room temperature for 2.0 hours. The mixture was diluted with HsO, filtered through a 0.4 µm polycarbonate membrane and the filtrate was evaporated in vacuo. The crude product was dissolved in a minimal amount of H2O and chromatographed on a 100 ml bed of HP-20 resin, elution being carried out with a linear gradient system of H2O / CH3CN. The product-containing fractions were combined and evaporated. The residue was taken up in H2O, filtered through a polycarbonate membrane and lyophilized to give 93 mg (yield of 63% based on the weight of the hydrate) of the desired lithium salt in the form of a white solid,

Thin layer chromatography CH2CI2-GH3OH-HOAC (8: 1: 1) Rf = 0.51, UV and APM,

Composition calculated for CigHtìOzCfeFLiP + 1.38 moles of water (MW, 495.94): G, 46.01; H, 4.42; F, 3.83; Cl, 14.30; P, 6.24.

Composition found: C, 46.10; H, 4.49; F, 3.82; Cl, 14.32, P, 6.43.

1 H NMR (400 MHZ):

O 11

81.53 ppm (3H, d, -OP {CH) CH -, J H-p = 14.6 Hz)

3 2

O 11

1.87-2.10 (2H, m, -OP (OCH) CH -J

3 2

2.27 (1H, dd, -CH (0H) CH2C02Li, Jh-H = 8.4 Hz, Jp_h = 1.1 Hz)

2.38 (1 H, dd, -CH (0H) CH2C02Li, JH-H = 4.7 Hz, Jh-p = 1.1 HZ)

4.29 (1H, m, -CH2CH (0H) CH2C02Li)

5.16-5.18 (4H, m, ArCH2OP and F-PhCH20) -7.11-7.52 (6H, m, aromatic)

Example 7

Mono-lithium salt of (S) acid [[[fluoro-4'trimethyl-3,3 ', 5 [biphenyl-1,1'] yl-2 methyl] amin] methoxyphosphinyl] -4 hydroxy-3 butyric acid

A. Fluoro-4'trimethyl-3,3 ', 5 [1,1' biphenyl] - 2 carboxylic acid

A solution of the aldehyde C (2) of Example 1 (1.0 g, 4.13 mmol) in acetone (10.0 ml) at 0 ° C (ice bath) was treated dropwise dropwise with 8.0 N Jones reagent (4.1 ml, excess) and the brown-green suspension obtained was mixed overnight under argon at room temperature. The excess oxidant was destroyed by adding isopropanol (10.0 ml) and the precipitated chromium salts were removed by a 1/4 inch layer of cellite. The filtrate was evaporated and taken up in EtOAc, then washed with 1 N HCl (2 times), a saturated NH4Cl solution (2 times) and a salt solution, then dried over anhydrous Na2SO4 and evaporated to give 1.011 g of '' a green solid with a melting point of 153-154 ° C.

The crude acid was purified via the dicyclohexylamine salt. Dicyclohexylamine (DCHA) (823 µl, 1.0 eq) was added to a solution of crude acid in EtOAc (5.0 ml). The solution was diluted with hexane and the crystallized salt was collected to give 997 mg (55% relative to the aldehyde, melting point 181-183 ° C) of the desired product in the form of a whitish and crystalline salt of DCHA.

The free acid was regenerated from DCHA by taking up the salt in a mixture of a 5% solution of KHSO4 and EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SO4 and evaporated in vacuo to give 554 mg (52% based on the aldehyde) of the desired acid.

Thin layer chromatography CH2CI2-CH3OH (9: 1) Rf = 0.37, UV and APM.

B. Fluoro-4'triméthyi-3,3 ', 5 [biphenyl-1,1'] - 2 carboxamide

A suspension of acid A (554 mg, 2.14 mmol) in dry CH2Cl2 (6.0 ml) and dry DMF (1 drop) at 0 ° C (ice bath) was treated dropwise using a syringe with oxalyl chloride (205 jxl. 2.35 mmol, 1.1 eq) and the light yellow solution was stirred under argon at temperature

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CH 676 984 A5

room for one hour. The mixture was evaporated in vacuo, removed with benzene (2 times) and dried in vacuo to give an acid chloride in the form of a yellow oil.

A cooled mixture (0 ° C, ice bath) of THF (3.0 ml) and concentrated NH4OH (2.0 ml, excess) was treated dropwise with a solution of crude acid chloride in THF (3.0 ml). The bright orange solution obtained was stirred under argon for 1.0 hour. This solution was taken up with a mixture of H2O and EtOAc and the organic phase was washed with saturated NaHGOs solution, then with H2O and with a saline solution to finally be dried over anhydrous Na2SO4 and evaporated, which gave 528 mg (96.1%) of crude amide as a light orange solid. Recrystallization from an EtOAc-hexane system provided 435 mg (79.1%) of purified amide in the form of yellow needles having a melting point 197-1989G.

ET20-acetone thin layer chromatography (1: 1) Rf = 0.83, UV and AMP

C. Fluoro-4 'trimethyl-3,3', 5 [l, biphenyl] -2 methanamine

A cooled solution (0 ° C, ice bath) of dry THF (5.0 ml) was treated with solid LÌAIH4 (125 mg, 3.3 mmol) and the gray suspension treated dropwise in five minutes with a solution of amide B (424 mg, 165 mmol) in THF (5.0 ml). The suspension obtained was stirred at room temperature under argon for 2.5 hours and brought to reflux for 45 minutes. The mixture was cooled to 0 ° C (ice bath) and the reaction blocked by the successive addition of 125 µl H20,125 µl 15% NaOH and 375 µl H2O. The precipitated aluminum salts were removed by filtration over anhydrous Na2SO4 disposed over a compact layer of cellite. The clear filtrate was evaporated in vacuo to give crude amine as a clear oil.

Et20-acetone thin layer chromatography (7: 3) Rf = 0.60, UV and APM. The amine was purified by the formation of its salt with HCl.

A solution of the crude amine (about 1.65 mmol) in absolute ethanol (8.0 ml) was treated with concentrated HCl (152 µl, 1.82 mmol) and the mixture was stirred for 15 minutes at room temperature under argon. The mixture was evaporated in vacuo to give a white crystalline solid. The solid was triturated in cold Et2O, separated by filtration and dried under vacuum to give 426 mg (92.4%) of the desired amine-HCl complex as white crystals.

D. (S) acid methyl ester - [[[[fluoro-4 'trimethyl-3,3', 5 [biphenyl-

1,1 '] yt-2] methyl] amino] methoxyphosphinyl] -4 t-butyldiphenyljsilyloxy-3-butyric

A solution of the methyl ester E (6) of Example 1 (about 2.0 mmol) in dry GH2Cl2 (5.0 ml) was treated with distilled trimethylsilyl diethylamine (758 µl, 4.0 mmol , 2.0 eq) and the clear mixture was stirred at room temperature under argon for one hour. The mixture was evaporated in vacuo, removed with benzene (1 time 15 ml) and dried in vacuo.

A cooled (0 ° C) solution of crude silylphosphonate in dry CH2CÌ2 (7.0 ml) and DMF (1 drop) was treated dropwise with distilled oxalyl chloride (192 µl, 2.2 mmoles, 1.1 eq). Gas emission occurred in the light yellow mixture. The solution was stirred at room temperature for one hour, evaporated in vacuo, stripped with benzene (2 x 15 ml) and dried in vacuo to give the crude phosphonochloridate as a yellow, viscous oil.

A cooled solution (0 ° C) of the phosphonochloridate and the biphenylamine complex. HCl C (416 mg, 1.49 mmol) in dry CH2Cl2 (10 ml) was treated with Et3N (641 d, 4.6 mmol, 2.3 eq) and 4-DMAP (24 mg, 0 , 2 mmol, 0.1 eq) and the clear mixture was stirred overnight under argon at room temperature. The mixture was taken up in a mixture of EtOAc and a 5% solution of KHSO4 and the organic phase was washed with brine, dried over anhydrous Na2 $ 04 and evaporated in vacuo to give 1.19 g of a yellow oil. The crude product was purified by flash chromatography on silica gel (60 g) eluting with hexane-acetone (7: 3). The product-containing fractions were evaporated to give 588 mg (59.5%) of the desired phosphonamide in the form of a pale yellow, viscous oil.

Hexane-acetone thin layer chromatography (7: 3) Rf = 0.20, UV and APM,

E. (S) - [[[[4-trimethyl-trimethyl-3,3 ', 5 [biphenyi-1,1'] yl-2lmethyl] amino] methoxyphosphinyi] -4 hydroxy-3 butyric acid methyl ester

A solution of silyl ether D (588 mg, 0.888 mmol) in dry THF (10.0 ml) was treated with glacial HOAc (203 nl, 3.55 mmol, 4.0 eq) and with a solution 1, 0 M tetrabutylammonium fluoride in THF (2.66 ml, 2.66 mmol, 3.0 eq) and the solution obtained was stirred overnight under argon at room temperature. The mixture was poured onto cold H2O and extracted with EtOAc. The organic phase was washed with saturated NaHCO3 solution and with brine, dried over anhydrous Na2SO4 and evaporated in vacuo to give 605 mg of an orange oil. The crude product was purified by flash chromatography on silica gel (36 g), elution being carried out with hexane-acetone (1: 1). The product-containing fractions were combined and evaporated to give 196 mg (50.4%) of the desired alcohol as a light orange oil.

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CH 676 984 A5

Hexane-acetone thin layer chromatography (1: 1) Rf = 0.16, UV and APM.

F. Acid mono-lithium salt (S) - [[[[fluoro-4 'trimethyl-3,3', 5 [biphenyl-1,11yl-2] methyl] amino] methoxyphosphinyl] -4 hydroxy- 3 butyric

A solution of diester E (105 mg 0.240 mmol) in dioxane (2.0 ml) was treated with a 1.0 N solution of LiOH (288 µl, 1.2 eq) and the white suspension was stirred under argon at room temperature for 4.0 hours. The mixture was diluted with H2O, filtered and the filtrate evaporated in vacuo. The residue was chromatographed on HP-20 resin (100 ml bed), elution being carried out with a linear gradient H2O / CH3CN. The product-containing fractions were combined and evaporated. The residue was taken up in H2O (50 ml), filtered through a 0.4 µm polycarbonate membrane and lyophilized to give 70 mg (62.7% based on the weight of the hydrate) of the lithium salt sought as a white solid.

Thin layer chromatography CH2CI2-CH3OH-HOAC (20: 1: 1) Rf = 0.19 UV and APM.

Composition calculated for C21H26NO5PFLÌ + 2.41 moles of H2O (MW 472.75): C, 53.35; H, 6.57; N, 2.96; F, 4.02; P, 6.55.

Composition found: C, 53.35; H, 6.52; N, 2.98; F, 4.05; P, 6.59.

1 H NMR (400 MHz):

O II

81.79-1.97 ppm (2H, m, -P (OCH) CH -}

3 2

2.26-2.44 ppm (2H, m, -CH2COgLi)

2.29 (3H, s, methyl aromatic)

2.31 (3H, d, aromatic methyl in relation to fluorine, Jhf = 1.4 Hz)

2.47 (3H, methyl aromatic)

3.46 & 3.50 (3H, 2 doublets, 2 diastereomers, Jhp = 10.5 Hz)

Where he

3.96 (2H, m, -PhCH ÎJHP (OCH) -

4.17 (1H, m, (-CH2CH (0H) CH2C02Li)

6.84-7.21 (5H, m, aromatic protons)

Examples 8 to 20

In accordance with the procedures as set out above and in particular in the practical examples, the following additional compounds can be prepared

2

3

OH

11 i

X

R-P-CH_-C

■ C-CH -CO -R = 22 OH

(CH ì l 2 z Z

37

CH 676 984 A5

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60

CH 676 984 AS

Ex. No.

n X

1 4.

C-H ,,

<§> "

ch2-o

C H_ 2 5

1 NH CH.

LS. OK

"WHO IS IT -

:, H - cö

C = C

I

O

I I

-CH.

* 5 ^

OK

© Sa

i • t i-f.

CH.O

f- \ OVch „-s •

> —F ~ | t m n

HC

H

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CH 676 984 A5

2nd.

\ ± /

1 NH H

Free 21

(S) - (diisopropyloxyphosphinyl-4 [[(1,1-dimethyl ethyt) diphenylsilyl] oxy] -3-butyric acid methyl ester

The iodide 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 moles, 93.92 g, 113.37 ml) was added all at once and the reaction mixture was stirred at 155 ° C (heated oil bath) under argon for 16.5 hours . The mixture was then cooled to room temperature. The excess triisopropyl phosphite and volatile reaction products were removed by short path distillation (10 mm Hg), followed by Kugelrohr distillation (0.50 mm Hg, 100 ° C, 8 hours). The product obtained was purified by rapid chromatography [column 95 mm in diameter, 6 inches of Merck silica gel, eluent hexane - acetone - toluene (6: 3: 1), speed 2 inches per minute, fractions of 50 ml ] to give 17.68 g (33.96 mmol, yield 75%) of the desired isopropyl phosphonate in the form of a viscous and clear oil.

Chromatography on hexane-acetone-toluene silica gel (6.3: 1) Rf = 0.32.

1 H NMR (270 MHz, CDCI3)

7.70-7.65 (m, 4H)

7.45-7.65 (m, 6H)

4.57-4.44 (m, 3H)

3.59 (s, 3H)

2.94 and 2.88 (2xd, 1H J = 3.7 Hz)

2.65 and 2.60 (2xd, 1H J = 7.4 Hz)

2.24-1.87 (series of m, 2H)

1.19 and 1.12 (2xd, 12H J = 6.3 Hz)

1.01 (s, 9H)

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CH 676 984 A5

Example 22

(S) - (hydroxymethoxyphosphinyl) -4 [[(dlmethyl-) dicyclohexylamine salt (1: 1)

1.1 ethyl) diphenylsilyl] oxy] -3 butyric.

The isopropyl phosphonate of Example 21 (30.5 mmol, 10.66 g) was stirred under argon at room temperature in 80 ml of dry CH2Cl2. This solution was treated dropwise in 5 minutes with bis-trimethylsilyltrifluoroacetamide (BSTFA) (32.8 mmol, 8.44 g, 8.71 ml), then with trimethylsilyl bromide (TMSBr) in 10 minutes (51, 3 mmol, 7.84 g, 6.75 ml). After stirring for 20 hours at room temperature, the reaction was blocked by the addition of 200 ml of a 5% aqueous solution of KH-SO4, followed by vigorous stirring for 15 minutes. The aqueous phase was extracted 3 times with ethyl acetate. The organic extracts were combined, washed once with brine, dried over Na2SO4 and concentrated in vacuo. The residue was subjected to two azeotropic distillations with 50 ml of toluene. The precipitate formed was suspended in toluene and filtered. The filtrate was concentrated and the azeotropic distillation / filtration process repeated. The filtrate obtained was evaporated in vacuo and then a high vacuum was applied to it via a vacuum pump for 5 hours. The viscous and clear oil was stirred under argon and at room temperature in 50 ml of dry pyridine. This solution was treated with dicyclohexylcarbodlimide (DCC) (22.6 mmol, 4.65 g) added all at once, then with methanol (41.0 mmol, 1.31 g, 1.67 ml). After 20 hours of stirring at room temperature, this reaction mixture was filtered through a cellite layer placed in a sintered glass filter. The cellite was washed with ethyl acetate and the filtrates were combined to be evaporated in vacuo. The residue was redissolved in ethyl acetate and washed twice with a 5% aqueous solution of KHSO4 and once with brine. The organic extract was dried over Na2SO4, filtered, concentrated and subjected to two azeotropic distillations with toluene, resuspended in toluene and filtered. The filtrate obtained was concentrated again, subjected to azeotropic distillation, filtered and evaporated in vacuo. To the residue was applied a high vacuum via a vacuum pump for 6 hours. The phosphonate monoester was thus obtained in the form of a clear viscous liquid (10.2 g, yield greater than 100%). Thin layer chromatography nPrOH-NhUOH-HaO (7: 2: 1) Rf = 0.50. A high vacuum was applied to the phosphonate monoester (1.21 g) via a vacuum pump for 4 hours. The product obtained (1.16 g, 2.57 mmol) was dissolved in 10 ml of dry ethyl ether and treated dropwise with dicyclohexylamine (2.65 mmol, 0.481 g, 0.528 ml). The homogeneous solution obtained was left to stand at room temperature for 7 hours, which resulted in significant crystal formation. The mixture was then placed at -20 ° C for 16 hours, then it was warmed to room temperature and filtered. The crystals were washed with cold, dry ethyl ether. Then, a high vacuum was applied to them via a vacuum pump in the presence of P2O5 for 18 hours, followed by the application of a high vacuum at 45 ° C for 4 hours. There was thus obtained 1.25 g (1.98 mmol, yield 77%) of the dicyclohexylamine salt in the form of a white powdery solid whose melting point was 155-156 ° C.

Chromatography on silica gel (20% MeOH / CH2Cl2) Rf = 0.57.

IH NMR: (270 MHz, GDCI3)

5 7.71-7.65 (m, 4H)

7.40-7.32 (m, 6H)

4.02 (m, 1 H)

3.52 (s, 3H)

3.28 and 3.22 (m, 1H)

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)

Spec. Weight: 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 Composition calculated for Ó22H31OPSÌ.C12H23N:

C, 64.63; H, 8.61; N, 2.22.

Composition found: C, 64.51: H, 8.49; N, 2.18.

Claims (6)

Claims 1. Compound as an inhibiting agent for the enzyme 3-hydroxy-3-methyl-glutaryl coenzyme A reductase, designated below by the abbreviation HMG Co A having the group: 41 5 10 15 20 25 30 35 40 45 50 55 60 CH 676 984 A5 0 il -P-CH -CH-CH -CO- 1 2 »2 X OH I (CH) l 2 n Z where X is an -O- or a -NH-, n is equal to 1 or 2 and Z is a lipophilic group. 2. Compound according to claim 1 having the structure 0 ". R-P-CH CH-CH CO „| * 5- -1 <É- X 1 (CH) i 2 n Z 2 = OH where R is OH, lower aikoxy or substituted or unsubstituted lower alkyl and Rx is H or substituted or unsubstituted lower alkyl and where X is -O- or -NH-; n is 1 or 2; Z is a lipophilic group which is or - Alkyl where the dotted lines represent optional double bonds, where R1, R2, R2a and R2b identical or different are chosen from the group consisting of H, halogens, trifluoromethy substituted or unsubstituted lower alkyls, haloalkyls, phenyl group, substituted phenyls or ORy, where Ry is H , alkanoyl, benzoyl group, phenyl group, halophenyl, phenyl- (substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkyl, cinnamyl group, haloalkyl, allyl group, (cycioalkyl substituted or unsubstituted) - (substituted or unsubstituted lower alkyl), adamantyl- (substituted or unsubstituted lower alkyl) or substituted phenyl- (substituted or unsubstituted lower alkyl); where RS and R5 'are the same or different and are selected from the group consisting of H, substituted or unsubstituted lower alkyls42 5 10 15 20 25 30 35 40 45 50 55 60 65 CH 676 984 A5 killed and OH, one of the groups R5 and R5 'being present when the auguel carbon is bonded to a double bond, O ii R6 is a (substituted or unsubstituted alkyl) -C— or an arylCH2-, R6a is substituted or unsubstituted lower alkyl, hydroxy, oxo, halogen or trifluoromethy, and q is 1, 2 or 3; as well as its salts. 3. A compound according to claim 2, wherein X is an -O-, R is an aikoxy, and Z is 4. The compound of claim 2, wherein X is -NH-, R is an aikoxy and (CH2) n is CH2 and Z is 5. Compound according to claim 2 chosen from the following compounds: Methyl ester or mono-lithium salt of (S) acid - {[[fluoro-4 'trimethyl-3,3', 5 [biphenyl-1,1 '] yI-2] methoxy] methoxyphosphinyl] -4 hydroxy -3 butyric, (S) acid di-lithium salt - [[[fluoro-4'trimethyl- 3.3 ', 5 [1,1' biphenyl] yl-2] methoxy] hydroxyphosphinyl] -4 hydroxy-3 butyric, Mono-lithium salt of acid (3S) - [[[fluoro-4 'trimethyl- 3,3 ', 5 [biphenyI-1,1'] yl-2] methoxy] methylphosphinyl] -4 hydroxy-3 butyric, Mono-lithium salt of (S) acid - [[[2,4-dichloro [(fluoro- 4 phenyl) methoxy-6] phenyl] methoxy] methoxyphosphiny!] - 4 hydroxy-3 butyric, Acid (3S) di-lithium salt - [[[2,4-dichloro [(fluoro- 4 phenyl) methoxy-6] phenyl] methoxy] hydroxyphosphinyl] -4 hydroxy-3 butyric, Acid (3S) - [[2,4-dichloro [(4-fluorophenyl) 6-methoxy] phenyl] methoxy] methylphosphinyl] -4 3-hydroxybutyric acid or its methyl ester or Mono-lithium salt of (S) acid - [[[[[fluoro-4 'trimethyl- 3,3 ', 5 [biphenyJ-1,1'] yl-2] methyl] amino] methoxyphosphinyl] -4 hydroxy-3 butyric. 6. A hypocholesterolemic or hypolipidemic composition comprising a compound defined in claim 2 and a vehicle for this compound which is pharmaceutically acceptable. 43