FI89493B - Foerfarande Foer framstaellning av terapeutiskt aktiva, phosphorhaltiga inhibitorer av HMG-CoA-reducuktas och vid foerfarandet anvaendbar ny mellanprodukt - Google Patents

Description

1 89493

A process for the preparation of therapeutically active phosphorus-containing HMG-CoA reductase inhibitors and a novel intermediate useful in the process. This patent application is a continuation of U.S. Patent Application 109,680 (filed October 19, 1987) and is a continuation of U.S. Patent Application No. 053,281 (filed May 5, 2003). .1987).

This invention relates to a process for the preparation of novel phosphorus-containing compounds which lower the activity of 3-10 hydroxy-3-methylglutaryl-coenzyme A reductase and are thus useful in inhibiting cholesterol biosynthesis, and to novel intermediates used in the process.

F. M. Singer et al., Proc. Soc. Exper. Biol. Med. 15 102 (1959) 370, and F. H. Hulcher, Arch. Biochem. Biophys.

146 (1971) 422, have disclosed some mevalonate derivatives which inhibit cholesterol biosynthesis.

Endo et al. have disclosed in U.S. Patent Nos. 4,049,495, 4,137,322 and 3,983,140 a fermentation product which is effective in inhibiting cholesterol biosynthesis.

. . This product is termed compactin and has a complex mevalonolactone structure according to Brown et al. (J. Chem. Soc. Perkin I, 1976, 1165).

GB Patent 1,586,152 discloses a group of • · 25 synthetic compounds of the formula

CHTV

<r; 30 R2 e; ··; * 4

2 8949J

wherein E represents a direct bond, a C 1-6 alkylene bridge or a vinylene bridge and the different Rs denote different substituents. The activity reported in that GB patent is less than 1% of the activity of the compactin.

U.S. Patent No. 4,375,475 to Willard et al. Discloses blood cholesterol and lipid lowering compounds having the structure

E

L

R3 where A is H or methyl, E is a direct bond, -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH = CH-, and Rx, R2 and R3 are each H, halogen, C 1-4 alkyl, C 1-4 haloalkyl, phenyl, halogen, C 1-4 alkoxy, C 2-8 alkanoyloxy, C 1-4 alkyl or C 1-4 haloalkyl substituted phenyl, or OR 4 where R 4 is H, C 2-8 alkanoyl, benzoyl, phenyl, halogen phenyl, phenyl-C 1-3 alkyl, C 1-9 alkyl, Cinnamyl, C 1-4 haloalkyl, allyl, cycloalkyl-C 1-3 alkyl, adamantyl-Cx_3 -alkyl or (substituted phenyl) -C 1-3 alkyl, wherein each of the substituents may be halogen, C 1-4 alkoxy, C 1-4 alkyl or C 1-4 haloalkyl; As well as the corresponding dihydroxy acids formed by hydrolytic opening of the lactone ring, and pharmaceutically acceptable salts and C 1-3 alkyl, phenyl and (dimethylamino or acetylamino substituted) -C 1-6 alkyl esters of said acids; all said compounds having enantiomers having tetrahydropyranate I of the trans racemate shown in the above formula; i 3 89493 in section 4R configuration.

Patent application WO 84/02131 (PCT / EP83 / 00308) filed in the name of Sandoz AG, based on U.S. Patent Application 443,668 (filed November 22, 1982) and U.S. Patent Application 548,850 (filed November 4, 1983), disclosed heterocyclic compounds analogous to mevalonolactone and derivatives thereof having the formula

R K

\ 2 I

10 Vi l ΐΏζν * - * * = ^ 15 // J ^ - R4 wherein one of the substituents R and R0 is \ R * where R4 is hydrogen, C1-4 alkyl, C1-4 alkoxy (except t-butoxy) , trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy, R 5 is hydrogen, C 1-3 alkoxy, trifluoromethyl, fluoro, chloro, 1'-phenoxy or benzyloxy and R 5a is hydrogen, C 1-4 alkyl, C 1-2 alkoxy, fluoro or chlorine, provided that R 5 and R 5a are both hydrogen with R 4 being hydrogen, R 5a being hydrogen with R 5 being hydrogen, not more than. one of R 4 and R 5 is trifluoromethyl, at most one of R 4 and R 5 is phenoxy and at most one of R 4 and R 5 is benzyloxy; And another primary or secondary C 1-6 alkyl, C 3-6 cycloalkyl or phenyl- (CH 2) m -, wherein m is 1, 2 or 3, R 2 is hydrogen, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-6. 4-alkyl (excluding t-butoxy), trifluoromethyl, fluorine, chlorine, phenoxy or benzyloxy, and: R 3 is hydrogen, C 1-6 alkyl, C 1-4 alkoxy, trifluoromethyl, fluorine, chlorine, phenoxy or benzyloxy, for provided that R 3 is hydrogen, when R 2 is hydrogen, at most one of R 2 and R 3 is trifluoromethyl, at most one of R 2 and R 3 is phenoxy and at most one of R 2 and R 3 is benzyloxy, X is - (CH 2) n-, where n is 0, 1, 2 or 3, or -CH = CH- and R6 5 4 3 | 2 1

Z is -CH-CH2-C-CH2-C00H

10 OH OH

wherein R 6 is hydrogen or C 10 alkyl; and in the form of the free acid, in the form of a physiologically hydrolyzable and physiologically acceptable ester or 6-lactone or in the form of a salt thereof · GB Patent 2,162,179A discloses naphthyl analogs of Mevalonolactone useful as inhibitors of cholesterol biosynthesis and -20 kenne 25 2

R 1 wherein R x is C 1-3 alkyl and Z is a group corresponding to formula Z 1 or Z 2:

HV

-CHCH- CHCH0 C00R-

4 2i 2 '. · I I

OH OH 0 35 | <Z1> <Z2 »li! 89493 wherein in the first formula R7 is hydrogen, a hydrolysable ester group or a cation.

EP 164 698A discloses the preparation of lactones useful as high blood cholesterol lowering agents by treating the amide with an organic sulfonyl halide R 5 SO 2 X and then removing the protecting group Pr: R 1 R 1 10 Pr0 2 J 2 R 4 Η ° Ί ^ ° R 2! R2 wherein X is halogen.

Pr is a carbinol protecting group,

R 1 is H or CH 3, R 11 and R 4 are each independently H, C 1-6 alkyl or phenyl-C 1-6 alkyl, wherein the phenyl is optionally C 1-6 alkyl, C 1-6 alkoxy - or halogen-substituted, R 2 is a group corresponding to formula A or B: 0

I J

’25 3 R‘ CH ’J

1 H | ? g ΑιΑ / Π] r8 (A) lg (B)

. · 30 R

wherein Q is R6-C- or R6-CH- ': / OH3 wherein R6 is H or OH; : 35 R is H or CH 3; b, c and d are possible double bonds; 6 89493 R 7 is phenyl or benzyloxy, the ring being in each case optionally substituted with C 1-4 alkyl or halogen, and R 8 and R 9 are each independently C 1-4 alkyl or halo-5; and R 5 is C 1-6 alkyl, phenyl or mono- or di (C 1-4 alkyl) phenyl.

DE-A-3,525,256 (Paul Leroy Anderson) discloses naphthyl analogs of mevalonolactone having the structure

HAr ^ Y0H

_ 1 «S sz -CHCH-CHCH-CO-R7 '' I

15 r ^ YVC A 2 A 2 ° V

I Π I H OH OH l

Q

Q 'wherein R 1 is alkyl, Z is a group corresponding to Q or Q 1 and R 7 is H or a hydrolyzable ester group; and which are useful as inhibitors of cholesterol biosynthesis as well as in the treatment of atherosclerosis.

Patent application WO84 / 02903, filed in the name of Sandoz AG and based on U.S. Patent Application No. 460,600 (filed January 24, 1983), discloses compounds analogous to mevalonolactone and useful as blood cholesterol lowering agents having the structure - R1 R 4 wherein both R 0 together form a group of formula 35 89493 8 7 6 5 "- C = C - C = C- or - (CH? Κ

ι I J

R 3 wherein R 2 is hydrogen, C 1-4 alkyl, C 1-4 alkoxy (except t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy, and R 3 is hydrogen, C 1-3 alkoxy, trifluoromethyl, fluoro , chlorine, phenoxy or benzyloxy, provided that at most one of R 2 and R 3 is trifluoromethyl, at most one of R 2 and R 3 is phenoxy and at most one of R 2 and R 3 is benzyloxy; R 3 is hydrogen, C 1-4 alkyl, fluoro, chloro or benzyloxy, R 4 is hydrogen, C 1-4 alkyl, C 1-4 alkoxy (excluding t-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy, R 5 is hydrogen, C 1-3 alkyl, C 1-4 alkoxy, trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy, provided that at most one of R 4 and R 5 is phenoxy and at most one of R 4 and R 5 is benzyloxy, R 5a is hydrogen, C 1-6 alkyl, C 1-6 alkoxy, fluorine or chlorine, 25 Ηχ / (CH 2) q

X is - (CH2) - or C = C

/ X (CH2) q H

wherein 30 n is 0, 1, 2 or 3 and both qs are 0 or one is 0 and the other is 1, and R 6 54 3 | 2 1

35 7. is -CH-CH, -C-CH2-COOH

OH OH

8 6 9 4 y 3 wherein R 6 is hydrogen or C 1-6 alkyl, with the general proviso that the phenyl group containing -X-Z and R 4 is in the ortho position relative to each other; and in the form of the free acid, in the form of its physiologically hydrolyzable and physiologically acceptable ester or δ-lactone or in the form of its salt.

EP-A-127 848A (Merck & Co., Inc.) discloses derivatives of 3-hydroxy-5-thiaarylalkanoic acids having the structural formula HO 0 I ox 1 C ^ S (O) L 5 I n

E

z where Z is 20 A <λ /

V

R 5 wherein R 1, R 2 and R 3 are each independently, for example, hydrogen, chlorine, bromine, fluorine, C 1-6 alkyl, phenyl, substituted phenyl or R 7, wherein R 7 is, for example, hydrogen, C 2-8 alkanoyl, benzoyl, phenyl, substituted phenyl, C 1-4 alkyl, cinnamyl, C 1-4 haloalkyl, allyl, cycloalkyl-C 1-3 alkyl, adamantyl-C 1-3 alkyl or phenyl-C 1-4 alkyl, and R 4, R 5 and R 6 is each independently hydrogen, chlorine, bromine, fluorine or C 1-6 alkyl; 35 n is 0, 1 or 2, 199493 E is -CH2-, -CH2-CH2-, -CH2-CH2-CH2-, -CH = CH-CH2- or -CH2-CH = CH- and X is for example hydrogen, C 1-3 alkyl or alkali metal cation or ammonium.

5 These compounds have a high blood cholesterol-lowering effect due to their ability to inhibit 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-Coa) reductase, and antifungal activity.

FR Patent Application 2,956,393A (Sanofi SA; filed April 1, 1986) discloses 3-carboxy-2-hydroxypropanephosphonic acid derivatives, including salts, having the formula R2 0 or3

: ii X

15 R.OOC-CHo-C-CPh-P

OH 1 OR 4 wherein R 1 and R 2 are each independently H, lower alkyl or optionally substituted aryl or aralkyl; 20 and which are useful as blood lipid lowering agents.

These compounds are reported to lower cholesterol, triglycerides, and phospholipids more than meglutone.

EP-A-142 146A (Merck & Co., Inc.) discloses mevilonin-like compounds having the structural formula

«0 O

30 'ΓΎκ1

E

i

Z

35 wherein R 89 is, for example, hydrogen or C 1-4 alkyl, E is -CH 2 -CH 2 -, -CH = CH- or - (CH 2) r -, and Z is a group 0 5 1) R 7 - wherein X is -O- or -NR 9 -, wherein R 9 is hydrogen or C 1-3 alkyl, R 7 is C 2-8 alkyl and R 8 is hydrogen or CH 3; 2) R 10 4 -R 11 R 12 R 12 wherein R 10, R 11 and R 12 are each independently, for example, hydrogen, halogen or C 1-4 alkyl; or 3) This invention relates to a process for the preparation of phosphorus-containing compounds which inhibit 3-hydroxy-3-methylglutaryl-coenzyme A reductase enzyme (HMG-CoA reductase) and are thus useful as blood cholesterol lowering agents and which contain of Chapter 11 89493 0 il -p-ch2-ch-ch2-co- 1 2 «2

X OH

(CH 2) n

5 Z

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

As used herein, the term "hydrophobic anchor" refers to a lipophilic group that, when attached to the upper HMG-like side chain of the molecule via a suitable linking group "X", binds to a hydrophobic "pocket" of the enzyme that is not utilized in HMG-CoA substrate binding. results in increased potency compared to compounds where Z is H.

Thus, the invention relates to a process for the preparation of compounds of formula 0-R-P-CH2-CH-CH2-CO2Rx X OH (I) (CH2) n

Z

wherein R is OH, lower alkoxy or lower alkyl, X is -O- or -NH-, n is 1, R 1 is R 2

Z is a group iC3J

R2a wherein R1, R2 and R2a may be the same or different and are each independently H, halogen, lower alkyl, haloalkyl, phenyl, substituted phenyl or ORy, wherein Ry is H, alkanoyl, benzoyl, phenyl, halophenyl , phenyl-substituted lower alkyl, lower alkyl, cinnamyl, haloalkyl, allyl, cycloalkyl-substituted lower alkyl, adamantyl-substituted lower alkyl or (substituted phenyl) substituted lower alkyl, and

Rx is hydrogen or lower alkyl, both for the preparation of their salts, which is characterized by the process, and for the preparation of a silyl ether of the formula 0 9 10 Rc-P-CH2-CH-CH2-CO2-alkyl X 0 (VII) (CH2) C (CH 3) 3 Z C 6 H 5 c 6 H 5 wherein R c is lower alkyl or lower alkoxy, cleaving the silyl ether group to form an ester of formula 0

B

Rc-P-CH2-CH-CH2-CO2Rx X ÖH (VIII) (CH2) n z (Rx is alkyl) 25 and the ester is converted, if desired, to the corresponding salt of formula

O

} l 30 R-P-CH2-CH-CH2-CO2 alkali metal

X ÖH

(CH2) n (HATE) «

Z

35 wherein R is lower alkyl or lower alkoxy, an acid of formula 13 89493 0

• I

r-p-CH2-CH2-CO2H

X ÖH

(CH2) n (VII IB)

5 Z

to the corresponding salt of formula 0 10 alkali metal O-P-CH 2 -CH-CH 2 -CO 2 alkali metal

X ÖH

(CH2) n (VIIIC) z15 or an acid of formula 011 ho-p-ch2-ch-ch2-co2h X ÖH (VIIID)

Z

The terms "salt" and "salts" refer to the basic salts formed with the inorganic and organic bases of the compounds. Such salts include ammonium salts, alkali metal salts such as lithium, sodium potassium salts (which are preferred), alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases such as amine salts, for example di-cyclohexylamine, benzathine. , N-methyl-D-glucamine and hydrabamine salts, salts with amino acids such as arginine and lysine salts, and the like. Non-toxic pharmaceutically acceptable salts are preferred, although other salts are useful, for example, in isolating or purifying the product.

14 89493

Thus, compounds of formula I include compounds

O

il 5 R-P-CH2-CH-CH2-CO2Rx (IA)

0 ÖH

(CH 2) n

Z

10 and

O

n R-P-CH2-CH-CH2-CO2Rx (IB)

NH ÖH

(CH 2) n

15 Z

The terms "lower alkyl" and "alkyl" as used herein, either alone or as part of another group, refer to both straight and branched chain hydrocarbon groups containing from 1 to 12 carbon atoms in the normal chain, preferably from 1 to 7 carbon atoms, such as methyl, ethyl , propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-25 trimethylpentyl, nonyl, decyl, undecyl, dodecyl, their various branched isomers and the like, as well as groups containing a halogen substituent such as F, Br, Cl, I or CF3, or alkoxy, aryl, arylaryl, haloaryl -, cycloalkyl, hydroxyl, alkylamino, alkanoylamino, arylcarbonylamino, nitro, cyano, thiol or alkylthio substituents.

The term "cycloalkyl" as used herein as part of another group refers to saturated cyclic hydrocarbon groups containing from 3 to 12 carbon atoms, preferably from 3 to 8 carbon atoms, and including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. yl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, any of which may be 1 or 2 halogen, 1 or 2 lower alkyl groups, 1 or 2 lower alkoxy groups, 1 or 2 hydroxyl groups, 1 or 2 alkylamino groups, 1 or Substituted with 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 "lower alkoxy" refers to lower alkyl groups as defined above attached to an oxygen atom.

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

The term "halogen" as used herein means chlorine, bromine, fluorine, iodine and CF 3, of which chlorine and fluorine are preferred.

Preferred compounds are compounds of formula I having the following structure R 0 CH, H CH, v. · '/ 2 \ / \

P C C0, R

i: 25 X OH (II) v CH 2 z where R is OH or OL 1, R x is Li or H, X is -O- or -NH- and R 1 - V V 2 Z is a group R 2a wherein R 1 is alkyl and / or or halogen-substituted phenyl, and R 1 and R 2 'are the same and are halogens or lower alkyl.

16 89493

The process according to the invention for the preparation of the compounds of formula I is illustrated according to the following reaction sequence and the related description.

cf »H5 a

OSi-C (CH3 »3 R -P (OalkylE) 2 O

Ra-P-CH2-CH-CH2-CO2-alkyl-I-CH2-C-L'H26CD2a? or "A lower alkoxyl"

IV

1. (CH) -SiBr, CH Cl, m

Ra -P-CH -CH-CH - CO.alkyl IV 2-> 2 ° _ OU ξ>

Decomposition of the phosphoric ester 'Si-C (Clf-) by S \ ^ ^ V'5 C6H5% I a VA. - Il = lower alkyl if R was lower alkyl VH. - II3 = O, If R3 was lower alkyl V3 (bOH-1) CC · 0.1K ~ ° H) (Ub = lower alkyl) Rho-f-CH.-CH-CH

(ester oil) I

Si-C (CH 2) 3 C6H5 C6H5

VI

VI 1) (cocl) 2; CH 2 Cl 2 (acid chloride formation) O

jfl 2) Z- (CH 2) 2 -XH (condensation reaction) RC-γ-CH 2 CH-CH 2 O 2 a Ikyy 1 i VA. --- 3> X 5 (C2H5> 3N '™ ΛΙ' Λ, η Si-C (CH3, 1 7 \ c6h5 c6H5

VII

(RC lower alkyl or lower alkoxyl)

1) (n-C4H9) 4NF, CH2COOH, THF (silyl ether O

VII _ decomposition) R-P-CIL-CH-CHj-CO Rx 2) Oli, d (oxane (hydrolysis) * = -y 1 öii * 7H2> n Z I.

17 89493

As can be seen from the above reaction sequence, the compounds of formula I can be prepared by the Arbuzov reaction of iodide by heating iodide A

5 C6H5 O-Si-C- (CH3) 3 | C6H5 (A) I -CH2-C-CH2-CO2alkyl t

B

10

and phosphonite / phosphite III

Ra-P (Oalkyl) 2 (III)

Wherein Ra is lower alkyl or lower alkoxy, under standard Arbutsov reaction conditions and using conventional procedures to form phosphinate / phosphonate IV

20 O

li

Ra-P-CH2-CH-CH2-C02alkyyli

Oalkyl-OSi-C (CH3) 3 (IV) C6H5 ^ C6H5 Phosphinate / phosphonate IV is a novel compound and as such forms part of this invention.

The phosphorus ester is then decomposed by treating a solution of phosphinate / phosphonate IV in which the solvent is an inert organic solvent such as dichloromethane with sequentially bis (trimethylsilyl) trifluoroacetamide (BSTFA) and trimethylsilyl bromide to form an inert gas such as Ra in IV has been lower alkyl, i.e. compound 18 89493 0 Ι · lower alkyl-P-CH 2 -CH-CH 2 -CO 2 alkyl d) H 6 (VA) i

Si-C (CH,) 3 R / \ 3 3 5 c6H5 c6h5 or phosphinic acid VB if Ra in compound IV has been lower alkoxy, i.e. compound

10 O

n HP-CH 2 -CH-CH 2 -CO 2 alkyl 1; OH 0 (VB)

Si-C (CH3) 3 c6h5 c6h5 15

Compounds VA and VB are new intermediates. When phosphonic acid VB is obtained, it is esterified by treatment in dry pyridine with alcohol VC

20 Rb0H (VC)

wherein Rb is lower alkyl, and with dicyclohexylcarbodiimide and stirring the resulting reaction mixture under an inert atmosphere such as argon to form the phosphonic acid monoalkyl ester VI

O

RbPOP-CH2-CH-CH2-CO2alkyl OH 0 (VI) i 30 Si-C (CH3) 3 c6h ^ c6h5

The ester VI or phosphinic acid VA is then dissolved in an inert organic solvent such as methylene chloride, benzene or tetrahydrofuran (THF), and treated with triinethylsilyldiethylamine and mixture I; ; 19 8 9 4 93

stirred under an inert atmosphere such as argon, the mixture is evaporated and the residue is then dissolved in methylene chloride (or other suitable inert organic solvent). The resulting solution is cooled to about 0-25 ° C, treated with oxalyl chloride and then evaporated to give the crude phosphonochloridate product. The phosphonochloridate is dissolved in an inert organic solvent such as methylene chloride, benzene, pyridine or THF, and the solution is cooled to about -20 to 0 ° C and treated with Compound B.

Z - (CH2) n-XH

used in an amount such that the molar ratio of ester VI or acid 15 VA to compound B is about 0.5: 1 to 3: 1, preferably about 1: 1 to 2: 1, followed by triethylamine and a catalytic amount of 4-dimethylaminopyridine (DMAP ) to form the adduct VII

20 O

II

Rr-P ---- CH2 -CH-CH2-CO2alkyl X O (CH2) n Si-C (CH3) 3 (VII) z c6h5 c6h5 wherein Rc is lower alkyl or lower alkoxy.

The silyl ether is decomposed by treating compound VII in an inert organic solvent such as tetrahydrofuran, glacial acetic acid and tetrabutylammonium fluoride to form ester VIII.

0 il

Rc-P-CH2-CH-CH2-CO2Rxalkyl I *

X OH

35 (CH2) n (VIII) Z (Rx is alkyl) 20 89493

Ester VIII can then be hydrolyzed to the corresponding alkali metal salt or acid, i.e., a compound where Rx is an alkali metal or H, by treatment in dioxane, tetrahydrofuran, or another inert organic solvent under an inert atmosphere such as argon and at 25 ° C. at temperature with a strong base such as lithium hydroxide used in an amount such that the molar ratio of base to ester VIII is about 1: 1 to 1.1: 1 to form the corresponding alkali metal salt HATE

O

M

R-P-CH2-CH-CH2-CO2 alkali metal X OH (VIIIA) 15 I 2 n

Z

wherein R is lower alkyl or lower alkoxyl. Compound VIIIA can then be treated with a strong acid such as HCl to give the corresponding acid 20

VII IB

O

II

The R-P-CH, -CH-CH, -C0oH

12:22 (VIIIB)

25 X OH

Wn

Z

Ester VIII, wherein R is lower alkoxyl, can be converted to the corresponding di-alkali metal salt by treatment at 50-60 ° C with a strong base used in an amount such that the base and ester

The molar ratio of VIII is about 2: 1 to 4: 1 to form salt VIIIC

2i 8 9 493 0 alkali metal-P-CH - CH-CH «-CO_ alkali metal 1 2 z 2 2

X ÖH

l (VIIIC) (CH-) 5 I 2 n

Z

The dialkali metal salt VIIIC can be converted to the corresponding acid where R is OH by treatment with a strong acid such as HCl to give acid VIIID.

O

of

HO-P-CHP-CH-CH ^ H ^ -CO

x Sh (five) (CH?) 15 «2 n

Iodine starting material A can be prepared from bromide C OH

BR-CH2-CH-CH2-CO2alkyl (C)

20 / prepared using the procedures described in Tetrahedron Lett. 26 (1985) 295-17, which is dissolved in a solution of imidazole and 4-dimethylaminopyridine in dimethylformamide (DMF), and the resulting solution is treated with t-butyldiphenylsilyl chloride under an inert atmosphere such as argon to form silyl ether D

Cv6H5 / 6H5 \ / (D) 30o Si-C (CH3) 3

Br-CH2-CH-CH2CO2 alkyl

Silyl ether D dissolved in an inert organic solvent such as ... methyl ethyl ketone or DMF is treated with sodium iodide under an inert atmosphere such as argon to form iodide A.

22 8 9493

Compound B to be used as starting material

Z- (CH_) -XH (B) 2 n can be prepared as described below, depending on the meaning of 5 Z and X.

Compounds of formula B wherein Z is R1 "" - <ί,

R

and X is 0, i.e., compounds having the structure R 1 R 2a - / q 1 - (CH 2) n - ° H (B1) R 2

can be prepared by treating aldehyde E

R1 R2 * (O- (CH2) n-1 "CHO (E) 25 ^ R2 with a reducing agent such as lithium aluminum hydride or sodium borohydride.

Compounds of formula B wherein Z is a group R1 r2M2OV- 35 '' R2 and X is -NH-, i.e. compounds having the structure 1 · 23 89493 R1 R2a ^ q "C- (CH2) n -NH2 (B2) 5 R2

can be prepared by oxidizing aldehyde E dissolved in acetone, for example with Jones reagent, to form acid F

R1 R2a— (ÖV <CH2> n-rC00H (F, 15 which is suspended in methylene chloride and treated with oxalyl chloride to give the corresponding acid chloride, which is dissolved in an inert organic solvent such as tetrahydrofuran and treated with a mixture of 2Q 1a, which contains concentrated ammonium hydroxide in tetrahydrofuran to form an amide having the structure R1 25 R2i - ^ ^ _ (CH2) n.1-C-NH2 (G) R2 ..... 2

The amide G is then reduced to the corresponding amide B by treatment with a reducing agent such as lithium aluminum hydride.

24 89493

The compounds of formula I can be prepared as racemic mixtures, the resolution of which to give the preferred S-isomer can be carried out later. However, the compounds of formula I can be prepared directly in the form of the 5S isomers as described herein and in the working examples below.

The compounds of the invention are inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase and thus useful in inhibiting cholesterol biosynthesis, as demonstrated by the following experiments.

1) Rat liver HMG-CoA reductase

Rat liver HMG-CoA reductase activity is measured using a modification of the method described by Edwards et al. [P. A. Edwards et al., J. Lipid Red.

20 (1979) 40]. Rat liver microsomes are used as the source of the enzyme and enzyme activity is determined by measuring the conversion of 14C-HMG-CoA substrate to 14C-mevalonic acid.

a. Preparation of microsomes 2-4 Sprague-Dawley rats fed choleraamine and decapitated, the liver is removed and the livers are homogenized in 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 IU / ml).

The homogenate is centrifuged (16,000 x g) for 15 minutes at 4 ° C. The supernatant is removed and recentrifuged under the same conditions. After a second centrifugation at 16,000 x g, the separated supernatant is centrifuged at 100,000 x g for 70 minutes at 4 ° C. The microsomal pellets are resuspended in as little phosphate buffer A as possible (3-5 ml / liver) and homogenized in a glass-glass homogenizer. Dithiothreitol (10 mM) is added and the preparation is aliquoted, rapidly frozen in an acetone-carbonic acid-35 ice bath and stored at -80 ° C. The specific activity of the first micros89493 rosome preparation was 0.68 nmol mevalonic acid / mg protein / minute. b. Enzyme assay

The reductase is determined from 0.25 ml of a mixture containing the following ingredients in the declared final concentrations:

Potassium phosphate, pH 7.0 0.04 M

KCl 0.05 M

Sucrose 0.10 M

10 EDTA 0.03 M

Dithiothreitol 0.01 M

NaCl 3.5 M

Dimethyl sulfoxide 1%

Microsomal protein 50 - 200 pg

[14C] (DL-) HMG-CoA

(0.05 pCi, 30-60 mCi / mmol) 100 μΜ NADPH (nicotinamide adenine

dinucleotide phosphate) 2.7 mM

The reaction mixture is incubated at 37 ° C. Under the conditions shown, the enzyme activity increases linearly up to 300 pg of microsomal protein / reaction mixture and the linear increase lasts for 30 minutes. The usual incubation time in drug studies is 20 minutes, resulting in the use of 25 HMG-CoA substrate at 100 pM, which is twice the concentration required to saturate the enzyme under the conditions shown. NADPH is used in excess of 2.7 times the concentration required to achieve the maximum rate of enzyme reaction.

Standardized tests to evaluate inhibitors are performed according to the following procedure.

The microsomal enzyme is incubated for 15 minutes at 37 ° C in the presence of NADHP. DMSO medium 35 with or without test compound is added and the mixture is incubated for an additional 15 minutes at 37 ° C. The enzyme assay is initiated by the addition of 14C-HMG-CoA substrate. After incubation for 20 minutes at 37 ° C, the reaction is stopped by the addition of 25 μΐ of 33% KOH. 3H-Meva-5 ionic acid (0.05 pCi) is added and the reaction mixture is allowed to stand at room temperature for 30 minutes. Add 50 μΐ of 5 N HCl to convert mevalonic acid to the lactone. Bromophenol blue is added as a pH indicator to detect a sufficient decrease in pH. The lactone formation reaction is allowed to proceed for 30 minutes at room temperature. The reaction mixtures are centrifuged for 15 minutes at 2,800 rpm. The supernatants are transferred to glass columns with an inner diameter of 0.7 cm on top of poured Ag 1 -X8 anion exchange resin (Biorad, formate form; 2 g) and eluted with 2.0 ml of water. The first 0.5 ml of eluate is discarded and the next 1.5 ml is collected and both tritium and carbon-14 in 10 ml of Opti-fluor supernatant are discarded. The results are calculated as the amount of mevalonic acid generated in 20 minutes (nmol) and corrected to correspond to a 100% recovery of 20 tritium. The effects of the drugs are expressed as I50 values (the concentration of drug that causes a 50% reduction in enzyme activity) obtained from the combined dose-response data with a 95% confidence interval.

The conversion of drugs in the lactone form to the sodium salts is carried out by dissolving the lactone in DMSO, adding a 10-fold molar excess of NaOH to the solution and allowing the mixture to stand at room temperature for 15 minutes. The mixture is then partially neutralized (pH 7.5 to 8.0) with 1 N HCl and then diluted to an enzyme reaction mixture.

2) Cholesterol synthesis in freshly isolated rat hepatocytes

The ability of compounds having activity as inhibitors of 35 HMG-CoA to inhibit the incorporation of 14C-acetate li 27 89 493 into cholesterol in suspensions prepared from freshly isolated rat hepatocytes was investigated using the methods originally described by Capuzzi et al. [D. M. Ca-puzzi and S. Margolis, Lipids 6 (1971) 602].

5 A. Isolation of rat hepatocytes

Sprague-Dawley rats (180-220 g) are anesthetized with non-butol (50 mg / kg). The abdomen is opened and the first branch of the portal vein is tied. One occluding suture is placed directly in the distal portion of the abdominal hollow-Kimo and a cannula is placed in the portal vein between the suture and the first branch vein. The liver is perfused after the vena cava is severed to allow outflow with preheated (37 ° C) oxidized buffer A (HBSS containing no calcium or magnesium but containing 0.5 pmol EDTA / ml) at a rate of 20 ml / minute. The liver is further perfused with 200 ml of preheated buffer B (HBSS containing 0.05% bacterial collagenase). After perfusion with Buffer B, the liver is excised and decapsulated in 60 ml of Waymouth's medium, allowing the cells to disperse in the medium. Hepatocytes are isolated by 3 minute centrifugation at low speed (50 x g). Hepatocyte pellets are washed once with Waymouth's medium and counted and their viability determined by trypan blue exclusion. The viability of these hepatocyte-rich cell suspensions is normally 70-90%.

B. ^ C-acetate incorporation into cholesterol Hepatocytes are resuspended in incubation medium (1M) (Tris-HCl, 0.02 M, pH 7.4; KCl, 0.1 M; sodium citrate, 3.3 mM; nicotinamide, 6 .7 mM; NADP, 0.23 mM; glucose-6-phosphate, 1.7 mM) to a cell concentration of 5 x 10 6 cells / 2.0 ml).

Test compounds are normally dissolved in DMSO or DMSO-water (1: 3) and added to IM.

28 89 493 The final concentration of DMSO in IM is up to 1.0% and has no significant effect on cholesterol synthesis.

Incubation is initiated by the addition of 14C-acetate (58 mCi / mmol, 2 pCi / ml) and placement of cell suspensions (2.0 5 ml) in 35 mm tissue culture dishes; the incubation time is 2.0 hours at 37 ° C. After incubation, the cell suspensions are transferred to glass centrifuge tubes and centrifuged at 50 x g for three minutes at room temperature. The cell pellets are resuspended in 1.0 ml of water, lysed and placed in an ice bath.

Lipids are extracted essentially as described by E. G. Bligh and W. J. Dyer [Can. J. Bio-chem. and Physiol. 37 (1959) 911]. The lower organic phase is separated and dried under a stream of nitrogen and the residue is resuspended in 100 of chloroform-methanol (2: 1). The total sample is spotted onto thin layer plates (silica gel LK6D) and the plates are developed from hexane-ethyl ether-acetic acid (75: 25: 1). The plates are scanned using an automatic BioScan scanning-20 system. The radioactive tracer contained in the cholesterol peak (Rf 0.28) is determined and the result is expressed as the total amount of scintillation per peak and the percentage of tracer in the total lipid extract. In control cultures, the radioactivity contained in the cholesterol peaks is usually 800 to 1,000 scintillations / minute and the amount of tracer in the total lipid extract is 9 to 20%; the results are consistent with those reported by Capuzzi et al. (9% extracted tracer in cholesterol).

30 The effects of drugs (percentage inhibition of cholesterol synthesis) are determined by comparing the percentage tracer levels of cholesterol in comparator and treated cultures. Dose-response curves are generated from data from two or more studies 29,849,493 and results are expressed as 150 values with a 95% confidence interval.

3) Cholesterol synthesis in human skin fibroblasts The selectivity of a compound that results in a greater inhibitory effect on liver tissue would be a hallmark of an inhibitor of cholesterol synthesis. Therefore, in addition to studying cholesterol synthesis inhibitors in hepatocytes, the activity of these compounds as cholesterol synthesis inhibitors is also being studied in cultured fibroblasts.

(a) Cultures of human skin fibroblasts

Human skin fibroblasts are cultured in Eagle's mini-myrrh medium (EM) containing 10% fetal 15 bovine serum. In each experiment, trypsin is added to stock cultures to disperse the monolayer, counted, and cells are transferred to tissue culture dishes in 35 mm diameter wells (5 x 10 6 cells / 2.0 mL). The cultures are incubated in a room containing 5% CO 2 and 95% humid room-20 air at 37 ° C for 18 hours. Enzymes involved in cholesterol biosynthesis are activated by removing the serum-containing medium, rinsing the monolayers, adding 1.0 ml of EM containing 1.0% fatty acid-free bovine serum albumin, and incubating the cultures for an additional 24 hours.

b) Addition of β-acetate to cholesterol Activated fibroblast cultures are rinsed with ENEM100 (Earle's Minimal Nutrient Medium). Test compounds are dissolved in DMSO or DMSO-EM-30 (1: 3) is added to the cultures (final DMSO concentration in cell cultures is up to 1.0%) and the cultures are pre-incubated in a room containing 5% CO 2 and 95% humid room air. At 37 ° C for 30 minutes. After pre-incubation with drugs, 1-c "-n; yttrium acetate (2.0 pCi / ml, 58 mCi / mmol) 3b is added and the cultures are re-incubated for four hours. After incubation, the medium is removed and the monolayer (200 pg) is removed. cell protein / culture) is scraped into 1.0 ml of water The lipids contained in the lysed cell suspension are extracted into the chloroform-methanol mixture with hepatocytes as shown in Figure 5.

Inhibition of cholesterol biosynthesis is determined by comparing the percentage of tracer in the cholesterol peaks from the reference culture and the drug-treated culture. Results are expressed as I50-ar-10 butter and are obtained from dose-response curves pooled from two or more experiments. The 95% confidence interval for Ib0 values is also calculated from the combined dose-response curves.

The compounds of formula I may be used in compositions consisting of at least one compound of formula I in combination with a pharmaceutical medium or diluent. The pharmaceutical composition may be formulated using conventional solid or liquid media or diluents and pharmaceutical excipients of a type suitable for the desired route of administration. The compounds may be administered orally, for example, in the form of tablets, capsules, granules or powders, or may be administered in the form of parenterally injectable preparations containing from 1 to 200 mg of active compound per therapeutic dose. The dose to be given depends on the unit dose, the symptoms, and the patient's age and body weight.

The compounds of formula I can be administered in the same manner as those known compounds (such as Lova statin) which have been proposed for use as inhibitors of cholesterol biosynthesis in mammals such as humans, dogs, cats and the like. Thus, the compounds of formula I may be administered in an amount of about 4 to 200 mg, either as a single dose or in separate doses 1 to 4 times a day, preferably 4 to 200 mg, in sub-doses of 1 to 100 mg. mg, preferably 0.5 to 50 mg, 2 to 4 times a day or in a sustained release form.

The following working examples represent preferred embodiments of this invention. All temperatures are in degrees Celsius unless otherwise noted. Flash chromatography treatments were performed on either Merck 60 or Whatman LPS-1 silica gel. Reverse phase 10 chromatography treatments were performed on CHP-20 MCI gel resin supplied by Mitsubishi, Ltd.

Example E 1 (S) -4 - [[[4'-fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl-2-yl] methoxy] methoxyphosphinyl] -3-hydroxybutanic acid, monolithium salt A. N- (2,4-dimethylbenzylidene) benzeneamine [Reference: U.S. Patent 4,375,475 (Merck), p. 39]

A solution of freshly distilled 2,4-dimethyl-20-benzaldehyde (Aldrich; 6.97 mL, 50 mmol) and distilled aniline (Aldrich; 4.56 mL, 50 mmol) in dry toluene (80.0 mL) was refluxed for three hours. under argon in a bottle equipped with a Dean-Starck water separator. The mixture was cooled and then evaporated under reduced pressure to a yellow oil. The crude oil was purified by Kugelrohr distillation (0.5 mmHg, 160-180 ° C) to give 8.172 g (78.1%) of the desired title benzeneimine as a pale yellow oil which crystallized on standing to melt at low temperature. TLC (hexane-acetone, 4: 1): Rf = 0.67 and 0.77 (geometric isomers), UV and I2.

32 8 9493 Γ ^ "“ »yCr * - 5 [Oj0iCH3 _ CH3 J 2 [ref: U.S. Patent 4,375,475 (Merck), p. 3]

To a mixture of the benzenimine imine prepared in Part A (6.0 g, 28.7 mmol) in glacial acetic acid (144 mL) was added palladium (II) acetate (6.44 g, 28.7 mmol) and a bright red solid. , the homogeneous solution was refluxed under argon for 1 hour. The resulting cloudy mixture was filtered warm through a tightly packed layer of J-inch diatomaceous earth into water (900 mL). The precipitated orange solid was collected by filtration and dried under reduced pressure at 65 ° C with P 2 O 4 for 16 hours to give 10.627 g (85.5%) of the desired title palladium complex as an orange solid, m.p. 194-196 ° C ( 203-205 ° C for a recrystallized analytical sample in the literature).

C. 4'-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-carboxaldehyde 1) Bromine (4-fluoro-3-methylphenyl) magnesium {Reference: U.S. Patent 4,375,475 (Merck), pp. 37 and 3¾

The title Grignard reagent from Part C (1) was prepared by adding 5-bromo-2 to magnesium turnings (1.35 g, 55.4 mmol, 8.0 eq.) Stirred in dry Et 2 O (70.0 mL). -fluorotoluene (Fairfield Chemical Co .; 22.5 g, 60.9 mmol) dropwise at a rate sufficient to keep the reaction mixture refluxing. The reaction was started on an ultrasonic device. After the addition of bromide was complete, the mixture was stirred for 1 hour under argon at room temperature and refluxed for 15 minutes, after which it was allowed to cool to room temperature.

li 33 89493 2) 4'-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-carboxaldehyde

In a second flask, a mixture of the dipalladium complex prepared in Part B (3.0 g, 5.92 mmol) and triphenylphosphine (14.52 g, 55.4 mmol, 8.0 eq.) In dry benzene (100 mL) was stirred at room temperature under argon. under 30 minutes. To this solution, a Grignard reagent freshly prepared and filtered (glass wool filling) according to Part C (1) was then added in one portion via syringe, and the mixture was stirred for 1.5 hours at room temperature under argon. 6.0 N HCl (35 mL) was added and the mixture was stirred for an additional hour at room temperature, then filtered through tightly packed diatomaceous earth (1 inch layer). The filtrate was extracted with Et 2 O (250 mL) and the extraction solution was washed with saturated NaCl solution (2 x 100 mL), dried over anhydrous MgSO 4 and evaporated under reduced pressure to give 13.35 g of a viscous orange oil which crystallized on standing. The crude orange solid was purified by flash chromatography using silica gel (700 g) as adsorbent and hexane as eluent followed by hexane-Et 2 O (95: 5). The product fractions were evaporated to give 1.507 g (89.9%) of the desired title aldehyde as a pale yellow solid, m.p. 72-75 ° C (m.p. 73-74 ° C). TLC (hexane-Et 2 O, 95: 5): Rf = 0.40, UV and PMA.

D. 41 Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-methanol To cooled (0 ° C, ice bath), dry Et 2 O (15.0 mL) was added LiAlH 2. (259 mg, 6.82 mmol, 0.55 eq.) And a solution of the aldehyde prepared in Part C (3.0 g, 12.4 mmol) was added dropwise over 15 minutes to the resulting gray suspension. in dry Et 2 O (15 mL). The mixture was stirred under argon at room temperature for 30 minutes, after which it was cooled back to 0 ° C and 260 μl of water, 260 μl of 15% NaOH and 780 μl of water were added dropwise. The suspension was diluted with EtOAc and filtered through tightly packed diatomaceous earth (1/4 inch 5 layer) over anhydrous Na 2 SO 4 and the colorless filtrate was evaporated under reduced pressure to give 2.99 g (98.8%) of a white solid. . Trituration of the crude solid with cold hexane and drying under reduced pressure afforded 2.467 g (81.6%) of the desired title alcohol as a white solid, m.p. 102-103 ° C. TLC (hexane-EtOAc, 9: 1): Rf = 0.24, UV and PMA.

E. (S) -3- [2,1-dimethylethyl) diphenylsilyl] oxy] -4- (hydroxymethoxyphosphinyl) butanoic acid, methyl ester 1) (S) -4-Bromo-3-hydroxybutanoic acid, methyl ester 1a) / R - (R *, R *) 1-2,3,4-trihydroxybutanoic acid, calcium salt, hydrate [Ref: Carbohydate Research 72 (1979) 301-304] To a solution containing D-isoascorbic acid (44.0 g, 250 mmol) in water (625 ml), calcium carbonate (50 g) was added, the suspension was cooled to 0 ° C and 30% 2 2 ° 2 (100 ml) was gradually added. The mixture was stirred at 30-40 ° C (oil bath) for 30 minutes.

Activated carbon (Darco; 10 g) was added and the black suspension was heated on a steam bath until the evolution of Cl 2 ceased. The suspension was filtered through diatomaceous earth and evaporated under reduced pressure (bath temperature 40 ° C). The residue was dissolved in water (50 mL), the solution was heated on a steam bath, and CH 2 OH was added until the solution became cloudy. The precipitated gum was collected by filtration and air dried to give 30.836 g (75.2%) of the desired calcium salt as a white powder. TLC (iPrOH-NH 4 OH-1 O 2, 7: 2: 1): Rf = 35 0.19, PMA.

I:: 35 89493 1 b) / S- (R *, S *) J-2,4-Dibromo-3-hydroxybutanoic acid, methyl ester {Reference: K. Bock et al., Acta Scandinavica (B) 37 (1983) 341-344] The calcium salt (30 g) prepared in part (1) (a) was dissolved in 30-32% HBr acetic acid solution (210 ml), and the solution was stirred at room temperature for 24 hours. Methanol (990 ml) was then added to the brown solution and the mixture was stirred overnight. The mixture was evaporated to an orange oil, the residue was dissolved in CH 2 OH (75 ml), the solution was refluxed for 2 hours and evaporated. The residue was partitioned between EtOAc (100 mL) and water and the organic phase was washed with water (2x) and saturated NaCl solution, then dried over anhydrous Na 2 SO 4 and evaporated to give 22.83 g (90.5%). ) crude dibromide as a pale orange oil. TLC (EtOAc-hexane, 1: 1): Rf = 0.69, UV & PMA.

le) (S) -4-Bromo-3-hydroxybutanoic acid, methyl ester Reference: Same as in the preparation of compound (1) (b)]

To a solution of argon purged with dibromide (20.80 g, 75.4 mmol) and anhydrous NaOAc (21.0 g) in EtOAc (370 mL) and glacial acetic acid (37 mL) was added 5% Pd / C catalyst (1.30 g) and the black suspension were stirred under H 2 (1 atm) while monitoring the uptake. After 2 h the consumption was complete, the mixture was filtered through diatomaceous earth, the filtrate was washed with saturated NaHCO 3 and saturated NaCl, then dried over anhydrous magnesium sulfate and evaporated to give the crude dibromoester as a brown oil. The crude oil was combined with a second batch (starting from 36.77 g of dibromide) and distilled under reduced pressure to give 25.77 g of 35 (61.3%) of the desired title dibromoester as a clear oil boiling in the range of 79-80 ° C. (1.0 mmHg).

36 89493 TLC (EtOAc-hexane, 1: 1): Rf = 0.44, PMA. Elemental analysis: Calculated for C 9 H 9 Cl 2 Br: C, 30.48; H, 4.60; Br, 40.56 Found: C, 29.76; H, 4.50; Br, 39.86 5 2) (S) -4-Bromo-3 - [(1,1-dimethylethyl) diphenylsilyl] oxy] butanoic acid, methyl ester

To a solution of the bromohydrin prepared in Part E (1) (4.0 g, 20.4 mmol), imidazole (6.94 g, 5.0 eq.) And 4-dimethylaminopyridine (4-DMAP) (12 mg, 0.005 eq. .) in dry DMF (40 ml), t-butyldiphenylsilyl chloride (5.84 ml, 1.1 eq.) was added and the resulting homogeneous mixture was stirred under argon at room temperature overnight. The mixture was partitioned between 5% KHSO 4 and EtOAc and the organic phase was washed with water and saturated NaCl solution, dried over anhydrous MgSO 4 and evaporated to give 9.32 g (100%) of crude silyl ether as a clear viscose. as an oil. TLC (hexane-EtOAc, 3: 1): silyl ether Rf = 0.75, UV and PMA.

20.

3) (S) -4-Iodo-3 - [(1,1-dimethylethyl) diphenylsilyloxy] butanoic acid, methyl ester

To a solution of the crude bromide prepared in Part E (2) (9.32 g, 201 mmol) in methyl ethyl ketone (60 mL, dried over 4 A molecular sieves) was added sodium iodide (15.06 g, 100.5 mmol, 5.0 eq.) And the resulting yellow suspension was refluxed for 5.0 h under argon. The mixture was cooled, diluted with EtOAc and filtered and the filtrate was washed with dilute 2q NaHSO 4 (colorless) and saturated NaCl solution, then dried over anhydrous Na 2 SO 4 and evaporated under reduced pressure to give 10.17 g of a yellow oil. The crude oil was purified by flash chromatography on silica gel (600 g) 25 as adsorbent and hexane-Cl 2 Cl 2 (3: 1) as eluent. The product fractions were combined and evaporated to give 7.691 g of 37,8493 (74.2%, combined yield of both steps) of the desired title iodide as a clear, colorless, viscous oil. TLC (hexane-EtOAc, 3: 1): Rf = 0.75, UV and PMA. Note: parallel iodide product spots containing 5 bromide starting material.) 4} (S) -4- (diethoxyphosphinyl) -3- [771/1-dimethylethyl) diphenylsilyl] oxy7-butanoic acid, methyl ester

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 was distilled off under reduced pressure (0.5 mmHg, 75 ° C) to leave a yellow oil (ca. 8.0 g). The crude oil was purified by flash chromatography on silica gel (400 g) as adsorbent and hexane-acetone (4: 1) as eluent. The product fractions were evaporated to give 3.222 g (41.5%) of the desired title phosphonate as a clear, colorless, viscous oil. TLC (hexane-acetone, 1: 1): Rf = 0.51 UV and PMA. In addition, 2.519 g of the starting iodide prepared in 20 parts (3) were recovered (corrected yield 61.1%).

5) (S) -3- (7H-1,1-dimethylethyl) diphenylsilyl-7-oxy] -4-phosphonobutanoic acid, methyl ester

To a solution of the phosphonate (9.85 g, 20.0 mmol) prepared in part (4) in dry CH 2 Cl 2 (60 mL) was added sequentially bistrimethylsilyl trifluoroacetamide (BSTFA) (5.31 mL, 32.0 mmol, 1.6 eq.) And trimethylsilyl bromide (TMSBr) (6.60 mL, 50.0 mmol, 2.5 eq.) And the resulting clear mixture were stirred. was placed under argon at room temperature overnight. 5% KHSO 4 (80 mL) was added and the mixture was extracted with EtOAc. The aqueous phase was saturated with NaCl and re-extracted with EtOAc. The combined organic layers were washed with saturated NaCl solution, dried over anhydrous

Na 2 SO 4 and evaporated under reduced pressure to give 38,889,493 crude title phosphonic acid as a viscous oil. TLC (iPrOH-NH 4 OH-H 2 O, 7: 2: 1): Rf = 0.30, UV and PMA.

6) (S) -3- [7 (1,1-dimethylethyl) diphenylsilyl] · 5 oxy] -4- (hydroxymethoxyphosphinyl) butanoic acid, methyl ester

To the crude phosphonic acid from Part (5) (ca. 20.0 mmol) mixed with dry pyridine (25 mL) was added CH 2 OH (dried over 3 A Mole-10 sieves; 1.62 mL, 40 mL). .0 mmol, 2.0 eq.) And dicyclohexylcarbodiimide (DCC) (4.54 g, 22.0 mmol, 1.10 eq.) And the resulting white suspension were stirred under argon at room temperature overnight. The pyridine was removed under reduced pressure and then azeotroped with benzene (2 x 15 mL). The residual oil was dissolved in EtOAc and the solution was filtered, washed with 1.0 N HCl and saturated NaCl solution, dried over anhydrous Na 2 SO 4 and evaporated under reduced pressure to give 8.272 g of crude title ester as an oil containing a small amount of amount of precipitated dicyclohexylurea (DCU). TLC (iPrOH-NH 4 OH-H 2 O, 7: 2: 1): Rf = 0.60, UV and PMA.

F. (S) -4- {7H-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] methoxyphosphinyl-3- (5-butyl) diphenyl-2,5-silyl-7-butanoic acid , methyl ester

The crude phosphonic acid monoester from Part E (6.595 g, ca. 14.7 mmol) was dissolved in dry CH 2 Cl 2 (30 mL), to which was added distilled trimethylsilyldiethylamine (5.60 mL, 29.4 mmol, 2.0 eq). 30 and the mixture was stirred under argon at room temperature for 1 hour. The mixture was re-evaporated under reduced pressure with the aid of benzene (1 x 30 ml) and dried under reduced pressure. The pale yellow viscous oil was dissolved in dry CH 2 Cl 2 (30 mL) and DMF (dried on 4 A molecular sieves; 2 drops), the clear solution was cooled to -10 ° C (salt-ice bath) and added 39 89493 syringe dropwise distilled oxalyl chloride (1.41 mL, 16.2 mmol, 1.1 eq). There was a strong evolution of gas and the solution turned a deeper yellow color. The mixture was stirred under argon at -10 ° C for 15 ml-3 minutes, then stirred at room temperature for 1 hour. The mixture was evaporated under reduced pressure and again with the aid of benzene (1 x 30 ml) and dried under reduced pressure to give the crude phosphonochloride as a yellow oil.

To a solution of the crude phosphonochloridate (ca. 14.7 mmol) in dry Cl 2 Cl 2 (10 mL) was added dropwise a solution of the biphenyl alcohol prepared in Part D (2.06 g, 8.43 mmol) in dry pyridine ( 15 ml) and the resulting mixture was stirred at room temperature under argon for 16 hours.

The mixture was evaporated to dryness under reduced pressure and the residue partitioned between 5% KHSO 4 and EtOAc.

The organic phase was washed with saturated NaHCO 3 and saturated NaCl, then dried over anhydrous Na 2 SO 4 and evaporated under reduced pressure to give 8.290 g of a brown oil. The crude product was purified by flash chromatography using silica gel (370 g) as adsorbent and hexane-acetone (70:30) as eluent. The product fractions were combined and evaporated to give 3.681 g (66%) of the desired title phosphonate as a pale yellow oil. TLC (hexane-acetone, 3: 2): Rf = 0.59, UV and PMA.

G. (S) -4- [7 '"Fluoro" 3,3', 5-trimethyl (1,1'-biphenyl) -2-yl] methoxyphosphinyl] -3-hydroxybutanoic acid, 30 methyl ester

To a mixture of the silyl ether prepared in Part F (1.103 g, 1.66 mmol) in dry THF (20.0 mL) was added glacial acetic acid (380 μL, 6.64 mmol, 4.0 eq) and 1 .0 M tetrabutylammonium fluoride solution 35 (4.98 mL, 4.98 mmol, 3.0 eq.) And the resulting clear yellow solution were stirred at room temperature under argon overnight. The mixture was partitioned between cold water and EtOAc and the organic phase was washed with saturated NaHCO 3 and saturated NaCl, dried over anhydrous Na 2 O 4 and evaporated to a viscous yellow oil (1.174 g). The crude oil was purified by flash chromatography on silica gel (47 g) as adsorbent and CH 2 Cl 2 -acetone (85:15) as eluent. The product fractions were evaporated to give 679 mg (93.1%) of the desired title alcohol as a clear viscous oil. TLC (hexane-acetone, 1: 1): Rf = 0.41, UV and PMA.

H. (S) -4-, [774 1-Fluoro-3,3,5-trimethyl (1,1'-biphenyl) -2-yl] methoxy] methoxyphosphinyl] -3-hydroxybutanoic acid, monolithium salt

To a solution of the methyl 15 ester prepared in Part G (184 mg, 0.420 mmol) in dioxane (5.0 mL) was added 1.0 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 water, filtered through a 0.4 polycarbonate film and evaporated under reduced pressure. Residue 20 was dissolved in water (75 ml), frozen and lyophilized. The crude acid was dissolved in as little water as possible and chromatographed on a 100 ml CHP-20 resin layer eluting with a linear H 2 O-CH 2 CN gradient. The product fractions were evaporated, the residue dissolved in water (50 ml) and the solution filtered through a 0.4 polycarbonate film and lyophilized to give 174 mg (89.1% based on the mass of hydrate) of the desired title monolithium salt as a white solid. TLC (iPrOH-NH 2 OH-H 2 O mixture, 7: 2: 1): Rf = 0.58, UV and PMA.

Elemental analysis (C 12 H 12 O 3 PFL 1 + 1.95 mol H 2 O; M = 465.46): Calculated: C, 54.19; H, 6.26; F, 4.08; P, 6.65 Found: C, 54.19; H, 6.21; F, 4.29; P, 6.43 1 H-NMR (400 MHz): δ 41 89493 δ 1.74 - 2.08 ppm (2 H, m, -PO (OCH-,) CH 2 -) 2.30 (3 H, s, aromatic methyl) 2.32 (3 H, d, aromatic methyl-5 in the Οί position relative to fluorine, JHF = 2.2 Hz) 2.35 to 2.62 (2 H, m, -CH 2 CO 2 Li) 2.46 (3H, s, aromatic methyl) 3.57 & 3.63 (3H, 2 doublets, -OP (OCH 3) -, 10 2 diastereomers, J n = 10.3 Hp

Hz) 4.28 (1H, m, -CH 2 CH (OH) CH 2 CO 2 Li)

O

II

4.97 (2H, m, PhOH 2 OP (OCH 3) R) 6.87 - 7.25 (5 H, m, aromatic Hs)

Example 2 (S) -4- [4'-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] methoxy] hydroxyphosphinyl] -3-hydroxybutanoic acid, dilithium salt To a solution containing Example 1 diester (374 mg, 0.853 mmol) in dioxane (8.0 mL), 1.0 N LiOH (2.6 mL, 3.0 eq.) Was added and the mixture was heated at 50 ° C (oil bath ) under argon for 5 hours. A white precipitate formed. The mixture was diluted with water and filtered. The aqueous solution was extracted once with Et 2 O, filtered through a 0.4 polycarbonate membrane and evaporated under reduced pressure. The crude product was chromatographed on CHP-20 resin (100 ml layer) eluting with a linear H 2 O-CH 3 CN gradient. The product fractions were evaporated under reduced pressure, the residue dissolved in water (50 ml) and the solution filtered through a 0.4 polycarbonate film and freeze-dried to give 260 mg (67.1% based on the mass of hydrate) of the desired title dilithium salt as a white solid. TLC (PrOH-NH 4 OH-H 2 O, 7: 2: 1): Rf = 0.47, '35 UV and PMA.

42 89493

Elemental analysis (C 2 (H 2) 6 O 6 PFLi 2 + 1.77 mol H 2 O): Calculated: C, 52.88; H, 5.67; F, 4.18; P, 6.82 Found: C, 52.88; H, 5.26; F, 4.24; P, 6.43 1 H-NMR (400 MHz, CD 3 OD):

5 O

c H

O 1.69 ppm (2H, m, -OCCH 2 CH (OH) -) 2.26-2.42 (2H, m, CH 2 CO 2 Li) 2.30 (3H, s, aromatic methyl) 2.31 (3H, d, Aromatic methyl in the Qd position relative to F, JUT, = rir 1.9 Hz) 2.38 (3H, s, aromatic methyl) 4.22 (1H, m, -CH (OH) CH2 ~)

O

II

4.75 (2H, m, PhCH 2 OP -) 6.86-7.23 (5H, M, aromatic protons)

Example 3 (3S) -4 - [(4'-fluoro-3,31-5-trimethyl (1,1'-biphenyl) -2-yl] methoxy] methylphosphinyl] -3-hydroxybutanoic acid, monolithium salt A. (S) -4- (7-chloro) methylphosphinyl-3- (1,1'-dimethylethyl) diphenylsilyl] oxy7-butanoic acid, methyl ester

The title phosphinochloridate compound is prepared as described in the first three paragraphs of Example 6, Part B.

B. (3S) -4- (7β-Fluoro-3,3 ', 5-trimethyl (1,11-biphenyl) -2-yl) methoxy7-methylphosphinyl] -3- (t-butyl) diphenylsilyloxybutanoic acid, methyl ester (0 ° C, ice bath) to a solution of phosphinochloridate from Part A (ca. 2.2 mmol) and biphenyl alcohol (429 mg, 2.2 mmol, 1.0 eq.) Prepared in Example 1, Part C (2) in dry In CH 2 Cl 2 (10 mL), Et 2 N (4.25 μL, 3.04 mmol, 1.4 eq) and 4-DMAP (27 mg, 0.22 mmol) were added and the resulting orange the solution was stirred at room temperature under argon overnight, the mixture was partitioned between 5% KHSO 4 and EtOAc, and the organic layer was washed with saturated NaCl solution, dried over anhydrous Na 2 SO 4 and evaporated to give 1 The crude oil was purified by flash chromatography using LPS-1 silica gel (44 g) as the adsorbent and EtOAc-hexane (1: 1) as eluent, the product fractions were combined and evaporated to give to 298 mg (21%) of the desired title condensation product as a pale yellow oil. 460 mg of the starting biphenyl alcohol prepared in Example 1, Part C (2) were also recovered (corrected yield 67%). TLC (EtOAc-hexane, 1: 1): Rf = 0.18, UV and PMA.

C. (3S) -4- [7,4'-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] methoxy] methylphosphinyl] -3-hydroxybutane- acid, methyl ester

To a solution of the silyl ether prepared in Part B (298 mg, 0.46 mmol) in dry THF (6.0 mL) was added glacial acetic acid (110 μL, 1.84 mmol, 4.0 eq.) And 1 A 0 M solution of tetrabutylammonium fluoride in THF (1.43 mL, 3.1 eq.) And the resulting solution were stirred under argon at room temperature overnight. The mixture was partitioned between cold water and EtOAc and the organic phase was washed with saturated NaHCO 3 and saturated NaCl, dried over anhydrous Na 2 SO 4 and evaporated to a yellow oil (273 mg). The crude oil was purified by flash chromatography using LPS-1 silica gel (11 g) as adsorbent and hexane-acetone (3: 2) as eluent. The product fractions were combined and evaporated to give 150 mg (80%) of the desired title 30 alcohol as a viscous oil. TLC (hexane-acetone, 1: 1): Rf = 0.23, UV and PMA.

D. (3S) -4- [4,4-Fluoro-3,3,5-trimethyl (1,1'-biphenyl) -2-yl] methoxy] methylphosphinyl] -3-hydroxybutanoic acid, monolithium salt 35 To the solution containing the methyl ester prepared in Part C (150 mg, 0.367 mmol) in dioxane (3.0 mL) was added 1.0 N LiOH (0.44 mL, 1.2 eq.) and the resulting the white suspension was stirred at room temperature under argon for 2 hours. The mixture was diluted with water, filtered through a 4 polycarbonate film and evaporated under reduced pressure to give a colorless glass.

The crude product was dissolved in as little water as possible and chromatographed using HP-20 (100 ml layer) as an adsorbent and a linear gradient of N 2 O-CH 2 CN eluting with. The product fractions were evaporated, the residue dissolved in water (50 ml) and the solution filtered through a 0.4 polycarbonate film and lyophilized to give 130 mg (79% by weight of hydrate) of the desired title lithium salt as a white solid. TLC (CH 2 Cl 2 -CH 3 OH-HOAc, 8: 1: 1): Rf = 0.52, UV and PMA.

Elemental analysis (C 21 H 25 F 5 FlP + 1.73 mol H 2 O; M = 445.49): Calculated: 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 1 H-NMR (400 MHz):

20 O

C

o 1.49 ppm (3H, d, -OP (CH3) -, JH_p = 14.7 Hz)

O

B

1.83 - 2.0 (2H, m, P- (CH3) CH2-) 2.27-2.40 (2H, m, CH2CO2Li) 2.30 (6H, s, 2 aromatic methyls) 2.44 (3H, s, aromatic methyl) 4.26 (1H, m, -CH 2 CH (OH) CH 2 CO 2 Li)

O

I) 4.87 (2 H, m, ArCH 2 OP (CH 3) -) δ 6.90 - 7.20 (5 H, m, aromatic Hs)

Example 4 (S) -4- (N- (2,4-Dichloro-6- (7,4-fluorophenyl) methoxy) 7-phenyl] methoxy] methoxyphosphinyl] -3-hydroxybutanoic acid, monolithium salt 35 A. 2,4-Dichloro-6- [4-fluorophenyl] methoxy / benzal - dehyde [Reference: J. Med. Chem. 29 (1986) 167] l · 45 89493

A solution of 4,6-dichloro-2-hydroxybenzaldehyde (13.77 g, 72.5 mmol) in DMF (100 mL) was stirred and K 2 CO 3 (12.02 g, 87 mmol) was added. . This mixture was heated at about 70 ° C for 60 minutes, after which 4-fluorobenzyl bromide (11.7 mL) was added. The resulting solution was stirred at 70 ° C for 3.5 hours, then poured into ice water (1.5 L), and the precipitate was filtered off, washed with water and recrystallized from Et 2 O-petroleum ether. 17.88 g of 10 (83%) off-white crystals were obtained, m.p. 107-108 ° C.

B. 2,4-Dichloro-6- (T4-fluoro-phenyl) -methoxy] -benzene-methanol

To a cold (0 ° C, ice bath) dry Etov (10.0 mL) was added LiAlH 4 (158 mg, 4.16 mmol, 0.6 eq.), And to the resulting gray suspension was added dropwise a solution containing a portion of A prepared aldehyde (2.06 g, 6.93 mmol) in dry THF (10 mL). The mixture was warmed to room temperature and stirred for 1 hour under argon. The mixture was cooled back to 0 ° C (ice bath) and water (160 μl), 15% NaOH (160 μl) and water (475 μl) were added dropwise sequentially. The precipitated salts were removed by filtration through tightly packed diatomaceous earth (1/4 inch layer) over anhydrous Na 2 SO 4. The clear filtrate was evaporated to give 2.052 g (98.9%) of crude alcohol as white crystals. Trituration with cold hexane gave 1.892 g (91.2%) of pure title alcohol as a white crystalline solid with a melting point of 72-73 ° C. TLC (hexane-acetone, 4: 1): Rf = 0.31, UV and: - PMA.

Elemental analysis (C 1 H 2 Cl 2 N 2 O 2; M = 301.142): Calculated: C, 55.84; H, 3.68; Cl, 23.55; F, 6.31 Found: C, 55.97; H, 3.71; Cl, 23.42; F, 6.30 46 89493 C. (S) (4-CCCl2 · · 4-dichloro-1- [74-fluorophenyl) methoxy? phenyl7methoxy7methoxyphosphinyl / -3- (t-butyl) diphenylsilyloxybutanoic acid, methyl ester

To a solution of the methyl ester prepared in Example 1, Part E (6) (ca. 3.84 mmol) in dry CH 2 Cl 2 (10 mL) was added distilled trimethylsilyl-diethylamine (1.46 mL, 7.68 mmol, 2, 0 eq.) And the resulting solution was stirred at room temperature under argon for 1 hour. The mixture was re-evaporated under reduced pressure with the aid of benzene (1 x 20 ml) and dried under reduced pressure to give crude silylated phosphonic acid monomethyl ester as a colorless oil.

A solution of the crude ester (ca. 3.84 mmol) in dry CH 2 Cl 2 (10 mL) and dry DMF (1 drop) was cooled to -10 ° C (salt-ice bath) and distilled oxalyl chloride (368 μL) was added dropwise. 4.22 mmol, 1.1 eq). Gas evolution from a clear yellow solution appeared. The mixture was stirred at room temperature under argon for 1 hour and re-evaporated under reduced pressure with the aid of benzene (2 x 20 ml) to give the crude phosphonochloride as a viscous yellow oil.

To the crude phosphonochloridate (ca. 3.84 mmol) dissolved in dry CH 2 Cl 2 (10 mL) was added the alcohol prepared in Part B at 0 ° C (ice bath) (1.15 g, 3.84 mmol, 1.0 eq. ) followed by Et 2 N (805 μL, 5.76 mmol, 1.5 eq.) and 4-DMAP (47 mg, 0.384 mmol, 0.1 eq.) and the mixture was stirred at room temperature under argon. overnight · The mixture was partitioned between 5% KHSO 4 and EtOAc, and the organic phase was washed with saturated NaCl solution, dried over anhydrous Na 2 SO 4 and evaporated to give 3.197 g of a dark brown oil. The crude product was purified by flash chromatography on silica gel 35 (160 g) as adsorbent and hexane-EtOAc (7: 3) as eluent. The product fractions were combined and evaporated to give 594 g (21.1%) of the desired title phosphonate as a yellow oil. In addition, 688 mg of the starting alcohol prepared in Part B were recovered (corrected yield 52.4%). TLC (hexane-acetone, 1: 1): Rf = 0.29, UV and PMA.

5 D. (S) 4-Dichloro-6- ([4-fluorophenyl) methoxy] phenyl] methoxy7-methoxyphosphinyl] -3-hydroxybutanoic acid, methyl ester

To a solution of the silyl ester prepared in Part C (578 mg, 0.788 mmol) in dry THF (8 mL) was added glacial acetic acid (180 μL, 3.2 mmol, 4.0 eq.) Followed by A 1.0 M solution of n-Bu 2 NF in THF (2.36 mL, 2.36 mmol, 3.0 eq.) And the resulting pale yellow solution were stirred under argon at room temperature overnight.

The mixture was poured into cold water and extracted with EtOAc (2x). The organic phase was washed with saturated NaHCO 3 and saturated NaCl, dried over anhydrous Na 2 SO 4 and evaporated to give 625 mg of a yellow oil. The crude product was purified by flash chromatography using silica gel (31 g) as adsorbent and hexane-acetone (7: 3) as eluent. The product fractions were combined and evaporated to give 339 mg (86.9%) of the desired product. . the title alcohol as a clear, colorless viscous oil. TLC (hexane-acetone, 1: 1): Rf = 0.25, UV and PMA.

25 E. (S) -4- ([2,4-dichloro-6- [7-fluorophenyl) methoxy] phenyl] methoxy] methoxyphosphinyl] -3-hydroxybutanoic acid, monolithium salt

To a solution of the phosphonate from Part D (132 mg, 0.267 mmol) in dioxane (2.5 mL) was added 1.0 N LiOH (0.32 mL, 1.2 eq.) And the mixture was stirred under argon at room temperature for 4 h. .0 hours. A white precipitate was observed to form. The mixture was diluted with water and filtered and the filtrate was evaporated to dryness under reduced pressure. The residue was chromatographed on HP-20 resin (100 ml layer) eluting with a linear gradient of H 2 O-CH 2 CN. The product fractions were combined and evaporated, the residue was dissolved in water and the solution was filtered through a 0.4 polycarbonate film and freeze-dried to give 108 mg (79% by mass of hydrate) of the desired title lithium salt as a white solid. . TLC (Cl 2 Cl 2 -CH 2 OH-HOAc, 20: 1: 1): Rf = 0.41, UV and PMA.

Elemental analysis (C 19 H 18 7 Cl 2 Fl 2 P + 1.42 mol H 2 O; M = 511.72): Calculated: C, 44.59; H, 4.10; Cl, 13.86; F, 3.71; P, 6.05 Found: C, 44.22; H, 4.09; Cl, 13.91; F, 3.72; P, 6.11 1 H-NMR (400 MHz):

O

O 1.98 - 2.11 ppm (2 H, m, OP (OCH 3) CH 2 CH (OH) - 2.26 - 2.45 ppm (2 H, m, -CH (OH) CH 2 CO 2 Li) 3.63 & 3.62 (3H, 2 doublets, 2 diastereomers,

O

II

OP (OCH 3) CH 2 -, JHp-11 Hz) 4.23 (1H, m, (-CH 2 CH (OH) CH 2 CO 2 Li) δ 5.16 (2 H, s, F-PhCH 2 O) 5.24 (2 H, d, ArCH 2 OP, JHp = 6.2 Hz) 7.13 - 7.53 (6 H, m, aromatic Hs)

Example 5 (3S) -4- [(2,4-Dichloro-6- [74-fluorophenyl) methoxy] 25-phenyl] methoxy] hydroxyphosphinyl] -3-hydroxybutanoic acid, dilithium salt

To a mixture of the diester prepared in Example 4, Part D (210 mg, 0.424 mmol) in dioxane (4.0 mL) was added 1.0 N LiOH (1.30 mL, 3.0 eq.) And the resulting colorless solid was added. the solution was heated at 50 ° C (oil bath) under argon for 3.5 hours. After 15 minutes, a white precipitate was observed. The mixture was diluted with water and filtered, and the filtrate was evaporated under reduced pressure. The residue was dissolved in as little water as possible and chromatographed on HP-20 resin (100 ml layer) eluting with a linear gradient of H 2 O-CH 3 CN 49 89493. The product fractions were combined and evaporated. The residue was dissolved in water (50 ml) and the solution was filtered through a 0.4 μm polycarbonate film and lyophilized to give 175 mg (81% based on the mass of hydrate) of the desired title dilithium salt as a white solid. TLC (CH 2 Cl 2 -CH 3 OH-HOAc, 8: 1: 1): Rf = 0.07 / UV and PMA.

Elemental analysis (C 18 H 18 Cl 2 F 2 Cl 2 P + 1.70 mol H 2 O; M = 509.62): Calculated: C, 42.42; H, 3.84; F, 3.73; Cl, 13.91; P, 6.08 Found: C, 42.46; H, 3.90; F, 3.93; Cl, 13.42; Δ, 5.66 1 H-NMR (400 MHz):

15 O

II

1.73 - 1.92 ppm (2H, m, -OP (OLi) -CH 2 CH (OH) - 2.27 (1H, dd, -CH (OH) CH 0 CO-Li, J '= 8.8 Hz ) -N nn 2.39 (1H, dd, -CH (OH) CH 2 CO 2 Li, JHH = 4.4 Hz) 4.26 (1H, m, CH 2 CH (OH) CH 2 CO 2 Li) δ 5.08 (2 H, s, F-Ph-CH 2 OAr) 7.03 to 7.53 (6 H, m, aromatic Hs).

Example 6 (3S) -4- (CO- [4-Dichloro-6- (T4-fluorophenyl) methoxy] phenyl] methoxy] methylphosphinyl] -3-hydroxybutanoic acid, methyl ester A. (3S) - (Y1,1-Dimethylethyl) diphenylsilyloxy7-4 - (ethoxymethylphosphinyl) butanoic acid, methyl ester

A mixture of the iodide prepared in Example 1, Part E (3) (4.68 g, 9.18 mmol) in methyl diethoxyphosphine (Strem Chemicals; 5.0 g, 36.7 mmol) was heated at 100 ° C (oil bath ) For 2.5 hours and then at 150 ° C for an additional 3 hours under argon. A white precipitate slowly formed in the yellow solution. Excess phosphine was distilled off under reduced pressure (0.5 mmHg) and the crude product was purified by flash chromatography on silica gel using hexane-acetone (65:35) as adsorbent.

50 ö9493

The product fractions were combined and evaporated to give 1.590 g (38%) of the desired title phosphinic acid ester (mixture of diastereomers) as a clear viscous oil. TLC (hexane-acetone, 3: 2): Rf (2 diastereomers) = 0.19 and 0.22 UV and PMA.

B. (3S) (4- (2,2,4-Dichloro-6- [4-fluorophenyl) netoxy phenyl] methoxy] methylphosphinyl) -3- (t-butyl) phenylsilylbutanoic acid, methyl ester

To a solution of the phosphin-10-nic acid ester prepared in Part A (605 mg, 1.3 mmol) in dry CH 2 Cl 2 (6.0 mL) was added bis (trimethylsilyl) trifluoroacetamide (BSTFA) (280 μL). 05 mmol, 0.8 eq.) And trimethylsilyl bromide (TMSBr) (210 μL, 1.57 mmol, 1.2 eq.) And the resulting solution was stirred at room temperature under argon overnight. 5% KHSO 4 (15 mL) was added and the mixture was extracted with EtOAc. The organic phase was washed with saturated NaCl solution, dried over anhydrous Na 2 SO 4 and evaporated under reduced pressure to give crude phosphinic acid as a colorless oil.

To a solution of crude phosphinic acid (ca. 1.3 mmol) in dry CH 2 Cl 2 (6.0 mL) was added distilled trimethylsilyldiethylamine (270 μL, 1.44 mmol, 1.1 eq.) And the resulting clear the mixture was stirred at room temperature under argon for 1 hour.

The mixture was re-evaporated from benzene under reduced pressure.

(1 x 15 ml) and dried under reduced pressure.

To a cooled (0 ° C, ice bath) solution of crude silylated phosphinic acid (ca. 1.3 mmol) in dry CH 2 Cl 2 (6.0 mL) and DMF (1 drop) was added dropwise distilled oxalyl chloride ( 130 [mu] l, 1.44 mmol, 1.1 eq). Gas evolution occurred. The mixture was stirred at room temperature under argon for 1 hour and then re-evaporated under reduced pressure with the aid of benzene (2 x 15 ml) and dried under reduced pressure to give crude phosphinochloridate as a yellow oil.

51 89493 To a cooled (0 ° C, ice bath) solution of phosphinochloridate (ca. 1.3 mmol) and the alcohol prepared in Example 1, Part E (6) (392 mg, 1.3 mmol) in dry Cl 2 Cl 2 (6, 0 mL), Et 2 N (275 μL, 1.97 mmol, δ 1.5 eq.) And 4-DMAP (16 mg, 0.13 mmol, 0.1 eq.) Were added and the resulting the resulting mixture was stirred under argon at room temperature overnight. The mixture was partitioned between 5% KHSO 4 and EtOAc and the organic phase was washed with saturated NaCl solution, dried over anhydrous Na 2 SO 4 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) as adsorbent and hexane-EtOAc (3: 2) as eluent. The product fractions were combined and evaporated to give 266 mg (28.3%) of the desired title product as a clear colorless oil. 197 mg of starting alcohol were also recovered (corrected yield 57%).

C. (3S) -4 - ((2,4-Dichloro-6- (T4-fluorophenyl) methoxy) -phenyl] phenyl] methoxy] methylphosphinyl] -3-hydroxybutanoic acid, methyl ester

To a solution of the silyl ester prepared in Part B (275 mg, 0.38 mmol) in dry THF (6.0 mL) was added glacial acetic acid (90 μL, 1.53 mmol, 4.0 eq.) And 1.0 M in THF solution of tetrabutylammonium fluoride (1.2 mL, 3.1 eq.). The resulting solution was stirred under argon at room temperature overnight. The mixture was partitioned between cold ... between water and EtOAc and the organic phase was washed with saturated NaHCO 3 and saturated NaCl, dried over anhydrous NaSO 4 and evaporated to give 258 mg of a yellow oil. The crude product was purified by flash chromatography using LPS-1 silica gel as adsorbent and hexane-acetone (1: 1) as eluent. The product fractions were combined and evaporated to give 142 mg (77%) of the desired title alcohol as a clear colorless oil. TLC (hexane-acetone, 1: 1): Rf = 0.20, UV and PMA.

52 89 493 D. (3S) -b-iCCZ, 4-Dichloro-6- (T4-fluoro-phenyl) -methoxy-phenyl] -methoxy-methyl-phosphinyl] -3-hydroxybutanoic acid, monolithium salt

To a solution of the methyl 5 ester prepared in Part C (142 mg, 0.296 mmol) in dioxane (3.0 mL) was added 1.0 N LiOH (0.36 mL, 1.2 eq.) And the resulting white solid. the suspension was stirred under argon at room temperature for 2 hours. The mixture was diluted with water and filtered through a 0.4 polycarbonate film, and the filtrate was evaporated under reduced pressure.

The crude product was dissolved in as little water as possible and chromatographed on HP-20 resin (100 ml layer) eluting with a linear gradient of H 2 O-CH 3 CN. The product fractions were combined and evaporated. The residue was dissolved in water and the solution was filtered through a polycarbonate film and lyophilized to give 93 mg (63% by weight of hydrate) of the desired title lithium salt as a white solid. TLC (CH 2 Cl 2 -CH 3 OH-HOAc, 8: 1: 1): Rf = 0.51, UV and 20 PMA.

Elemental analysis (C 18 H 18 N 2 O 2 Cl 2 FlP + 1.38 mol H 2 O; M = 495.94): Calculated: C, 46.01; H, 4.42; F, 3.83; Cl, 14.30; P, 6.24 Found: C, 46.10; H, 4.49; F, 3.82; Cl, 14.32; P, 6.43 1 H-NMR (400 MHz): δ 53 89493 o

S 1.53 ppm (3 H, d, -OP (CH 2) CH 2 - J)

—3 ^ 2 'JH-P

14.6 Hz)

O

K

1.87 - 2.10 (2H, m, -OP (OCH 2) CH 2 -) δ 2.27 (1H, dd, -CH (OH) CH 2 CO 2 Li, JH-H = 8.4 Hz p = 1'1 Hz> 2.38 (1H, dd, -CH (OH) CH 2 CO 2 Li, 4.7 Hz, JH_p = 1.1 Hz) 4.29 (1H, m, -CH 2 CH (OH) CH 2 CO 2 Li) ) 5.16 - 5.18 (4 H, m, ArCH 2 OP and F-PhCH 2 Q) -)

7.11-7.52 (6 H, m, aromatic Hs)

Example 7 (S) -4- [7H-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] methyl] amino] methoxyphosphinyl-3-hydroxybutanoic acid, monolithium salt A 4-Fluoro-3,31,5-trimethyl (1,1'-biphenyl) -2-carboxylic acid

To a solution of the aldehyde prepared in Example 1, Part C (2) (1.0 g, 4.13 mmol) in acetone 20 (10.0 mL) at 0 ° C (ice bath) was added dropwise 8.0 N

Jones reagent (4.1 mL, excess) and the resulting brown-green suspension were stirred under argon at room temperature overnight. Excess oxidant was destroyed by the addition of isopropanol (10.0 mL) and the precipitated chromium salts were removed by filtering the mixture through diatomaceous earth (1/4 inch layer). The filtrate was evaporated, the residue was dissolved in EtOAc and the solution was washed with 1.0 N HCl (2x), saturated NH 4 Cl (2x) and saturated NaCl, then dried over anhydrous Na 2 SO 4 and evaporated to give 1.011 g of a green solid with a melting point of 153-154 ° C were obtained.

The crude acid was purified by preparing a dicyclohexylamine salt. To a solution of crude acid in EtOAc (5.0 mL) was added dicyclohexylamine (DCHA) (823, 1.0 eq). The solution was diluted with hexane and the precipitated crystalline salt was collected to give 997 g (55% based on the aldehyde) of the desired product as an off-white crystalline DCHA salt, m.p. 181-183 ° C.

The DCHA salt was converted to the title free acid by partitioning between 5% KHSO 4 and EtOAc. The organic phase was washed with saturated NaCl solution, dried over anhydrous Na 2 SO 4 and evaporated under reduced pressure to give 554 mg (52% based on the aldehyde) of the desired title acid.

TLC (CH 2 Cl 2 -CH 3 OH, 9: 1): Rf = 0.37, UV and PMA.

B, 41-fluoro-3,31,5-trimethyl (1,1'-biphenyl) -2-carboxamide

To a suspension of the acid prepared in Part A (554 mg, 2.14 mmol) in dry CH 2 Cl 2 (6.0 mL) and dry DMF (1 drop) was added dropwise at 0 ° C (ice bath) distilled water. oxalyl chloride (205 μL, 2.35 mmol, 1.1 eq) and the resulting clear yellow solution was stirred under argon at room temperature for 1 h. The mixture was evaporated under reduced pressure and again with benzene (2x) and dried under reduced pressure to give the crude acid chloride as a yellow oil.

To a cooled (0 ° C, ice bath) mixture of THF (3.0 mL) and concentrated NH 4 OH (2.0 mL, excess) was added a solution of crude acid chloride in THF (3, 0 ml) and the resulting bright orange solution was stirred at room temperature under argon for 1 hour. The mixture was partitioned between water and EtOAc and the organic phase was washed with saturated NaHCO 3, water and saturated NaCl, then dried over anhydrous Na 2 SO 4 and evaporated to give 528 mg (96.1%) of crude amide as a pale orange solid. . Recrystallization from EtOAc-hexane once gave 435 g (79.1%) of purified title amide as pale yellow needles with a melting point of 197-198 ° C. TLC (Et 2 O-acetone, 1: 1):

Rf = 0.83, UV and PMA.

55 89493 C. 4'-Fluoro-3,31,5-trimethyl (1,11-biphenyl) -2-methanamine To cooled (0 ° C, ice bath) dry THF (5.0 mL) was added solid LiAlH 2. a (125 mg, 3.3 mmol) and a solution of the amide prepared in Part B (424 mg, 1.65 mmol) in THF (5.0 mL) was added dropwise over 5 minutes to the resulting gray suspension. The resulting suspension was stirred at room temperature under argon for 2.5 hours and then refluxed for 10 to 45 minutes. The mixture was cooled to 0 ° C (ice bath) and 125 μl of water, 125 μl of 15% NaOH and 275 μl of water were added dropwise sequentially. The precipitated aluminum salts were removed by filtration through tightly packed diatomaceous earth over anhydrous Na 2 SO 4.

The clear filtrate was evaporated under reduced pressure to give the crude amine as a clear oil. TLC (Et 2 O-acetone, 7: 3): Rf = 0.60, UV and PMA. The amine was purified as the HCl salt.

To a solution of the crude amine (ca. 1.65 mmol) in absolute EtOH (8.0 mL) was added concentrated HCl (152 μL, 1.82 mmol) and the mixture was stirred at room temperature under argon for 15 minutes. The mixture was evaporated under reduced pressure to give a crystalline white substance. This solid was triturated with cold Et 2 O. ·. 25 and collected by filtration and dried to give 426 mg (92.4%) of the title amine hydrochloride as fine white crystals.

D. (S) -N- [CCCO4'-fluoro-3,31,5-trimethyl (1,1'-biphenyl) -2-yl] methyl] amino] methoxyphosphinyl-3- (t-butyl) -3-diphenylsilyloxybutanoic acid, methyl ester

To a solution of the methyl ester prepared in Example 1, Part E (6) (ca. 2.0 mmol) in dry CH 2 Cl 2 (5.0 mL) was added distilled trimethylsilyldiethylamine (758 μL, 4.0 mmol, 2.0 eq). .) and the resulting clear mixture was stirred at room temperature under argon for 1 hour. The mixture was re-evaporated under reduced pressure with the aid of benzene (1 x 15 ml) and dried under reduced pressure.

56 89 493 To a cooled (0 ° C) solution of crude silyl phosphonate in dry CH 2 Cl 2 (7.0 mL) and DMF (1 drop) was added dropwise distilled oxalyl chloride (192 μL, 2.2 mmol, 1.1 eq.). Gas was observed to evolve from the clear 5 yellow solution. The solution was stirred at room temperature for 1 hour and re-evaporated under reduced pressure with the aid of benzene (2 x 15 ml) and dried under reduced pressure to give the crude phosphonochloridate as a yellow viscous oil.

To a cooled (0 ° C) solution of phosphonochloridate and biphenylamine HCl prepared in Part C (416 mg, 1.49 mmol) in dry Cl 2 Cl (10 mL) was added Et 3 N (641 μL, 4 mL). , 6 mmol, 2.3 eq.) And 4-DMAP (24 mg, 0.2 mmol, 0.1 eq.) And the resulting clear yellow mixture were stirred at room temperature under argon overnight. The mixture was partitioned between 5% KHSO 4 and EtOAc and the organic phase was washed with saturated NaCl solution, dried over anhydrous Na 2 SO 4 and evaporated under reduced pressure to give 1.19 g of a yellow oil. The crude product was purified by flash chromatography using silica gel (60 g) as adsorbent and hexane-acetone (7: 3) as eluent. The product fractions were evaporated to give 588 mg (59.5%) of the desired phosphonamide of title 25 as a pale yellow viscous oil. TLC (hexane-acetone, 7: 3): Rf = 0.20, UV and PMA.

E. (S) -4-CCC & 1-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] methyl] amino] methoxyphosphinyl-3-hydroxybutanoic acid, methyl ester 30 To a solution containing Part D prepared silyl ether (588 mg, 0.888 mmol) in dry THF (10.0 mL), glacial acetic acid (203 μL, 3.55 mmol, 4.0 eq.) and a 1.0 M solution of tetrabutylammonium fluoride in THF ( 2.66 mL, 2.66 mmol, 3.0 eq) and the resulting solution was stirred under argon at room temperature overnight. The mixture was poured into cold water and extracted with EtOAc. Organic i.

The phase was washed with saturated NaHCO 3 and saturated NaCl, then dried over anhydrous Na 2 O 4 and evaporated under reduced pressure to give 605 mg of an orange oil. The crude product was purified by flash chromatography-5 using silica gel (36 g) as adsorbent and hexane-acetone (1: 1) as eluent. The product fractions were combined and evaporated to give 196 mg (50.4%) of the desired title alcohol as a pale orange oil. TLC (hexane-acetone, 1: 1): Rf = 0.16 UV 10 and PMA.

F. (S) -b-CCC & '- fluoro-3,3', 5-trimethyl (1,1'-biphenyl) -2-yl] methyl7amino] methoxyphosphinyl-3-hydroxybutanoic acid, monolithium salt

To a solution of the di-15 ester prepared in Part E (105 mg, 0.240 mmol) in dioxane (2.0 mL) was added 1.0 N LiOH (288 μL, 1.2 eq.) And the resulting white solid. the suspension was stirred under argon at room temperature for 4 hours. The mixture was diluted with water and filtered, and the filtrate was evaporated under reduced pressure.

The residue was chromatographed on HP-20 resin (100 ml layer) eluting with a linear water-CH 2 CN gradient. The product fractions were combined and evaporated. The residue was dissolved in water (50 ml) and the solution was filtered through a 0.4 μm polycarbonate film and freeze-dried to give 70 mg (62.7% based on the mass of hydrate) of the desired title lithium salt as a white solid. TLC (CH 2 Cl 2 -CH 3 OH-HOAc, 20: 1: 1): Rf = 0.19, UV and PMA.

... Elemental analysis (C 21 H 26 NO 5 PFLi + 2.41 mol H 2 O; M = 30 472.75): Calculated: C, 53.35; H, 6.57; N, 2.96; F, 4.02; ; P, 6.55 Found: C, 53.35; H, 6.52; N, 2.98; F, 4.05; P, 6.59 35 1 H-NMR (400 MHz): 58 89493 δ S 1.79 - 1.97 ppm (2 H, m, -P (OCH-δ) CH 2 -) 2.26 - 2, 44 ppm (2H, m, -C 2 CCl 2, 2.29 (3H, s, aromatic methyl) 2.31 (3H, d, aromatic methyl in the Qi position relative to fluorine, J n = 1.4 Hz) Γ1Γ 2.47 (3H, aromatic methyl) 3.46 & 3.50 (3H, 2 doublets, 2 diastereomers, JHp = 10.5 Hz)

O

3.96 (2H, m, -PhCH 2 HP (OCH 3) - 4.17 (1H, m, (-CH 2 CH (OH) CH 2 CO 2 Li) 6.84 - 7.21 (5 H, m, aromatic protons))

Examples 8-20 15. .

Following the procedures outlined above and described in the previous working examples, the following additional compounds can be prepared: 59 89493

Example

__! L_ _2_ _n_ X RX

8. OH (O) -fH2 1 0 H

cI <oC

8 · C2H50 CH3 ^ OV- 2 NH CH3 10 φ f3 L. ci 10 · C3H7 (of 10 Li 15 / - [cl \ 2 / C2H5

H · CH3 ° (5yCH2-0 2 NH H

20

S

12. OH ΊρΊ 2 0 H

(o ^ oX ": 30 13 · W <g> -c„ 2-o 1 0 “(pfcl 35 60 89493

First of all

No. R _Z_ _n_ _X_ RX

1.4. C5h11 1 NH CH3 5 c »2-f, 0ir 10 =» 3

15. OK C.H.-CH 2 0 OK

2 5 I

oo ch3 20 16 · 0 [te 2 0 or3

17. CH 3 O »H H

W 2 I I

- £ 0 "·

B

30 I: 61 89493

Pre-character no ° - —6 --- Ϊ —-- □ _ _JS_ R * C-H -C-C-0 18> CH, 7 \ ".

OH

10

19 · B0 <Ö> -, o U

15 fo) 20. CH O fr \

3> 2 / \ 2/1 ^ H

20 62 89493

Example 21 (S) -4- (Diisopropyloxyphosphinyl) -3 - [(1,1-dimethylethyl) diphenylsilyl] oxy] butanoic acid, methyl ester Iodide 5 (21.70 g, 45.1 mmol) prepared in Example 1, Part E (3) stirred under high vacuum for 30 minutes. Freshly distilled triisopropyl phosphite (93.92 g, 113, 37 mL, 0.451 mol) was added in one portion and the reaction mixture was stirred under argon and heated in a 155 ° C oil bath for 16.5 hours. The mixture was then cooled to room temperature. Excess triisopropyl phosphite and volatile reaction products were removed by short path distillation (10 mmHg) followed by Kugelrohr distillation (0.50 mmHg, 100 ° C, 8 hours). The product was further purified by flash chromatography 15 (6 inch column, 95 mm diameter; Merck silica gel; hexane-acetone-toluene, 6: 3: 1; flow rate 2 in. / Min, 50 mL fractions) to give 17.68 g (33.96 mmol, 75% yield) of the title isopropylphosphonate as a clear viscous oil. TLC 20 (silica gel; hexane-acetone-toluene, 6: 3: 1): Rf = 0.32 1 HNMR: (270 MH, CDCl 3) δ 7.70 - 7.65 (m, 4H) 7.45 - 7.35 (m, 6H) δ 4.57-4.44 (m, 3H) 3.59 (s, 3H) 2.94 and 2.88 (2x, 1H J = 3.7 Hz) 2, 65 and 2.60 (2 xd, 1H J = 7.4 Hz) 2.24 - 1.87 (set of m, 2H) 30 1.19 and 1.12 (2 xd, 12H J = 6.3) Hz) 1.01 (s, 9 H)

Example 22 (S) -4- (Hydroxymethoxyphosphinyl) -3- [7,1-dimethylethyl) diphenylsilyl] oxy7-butanoic acid, methyl ester, dicyclohexylamine salt (1: 1)

The isopropyl phosphonate prepared in Example 21 (10.66 g, 30.5 mmol) was stirred at room temperature.

e3 89493 under dry Cl 2 Cl 2 (80 mL). To this solution was added dropwise over 5 minutes bistrimethylsilyl trifluoroacetamide (BSTFA) (8.44 g, 8.71 mL, 32.8 mmol) followed by dropwise addition of 5 trimethylsilyl bromide (TMSBr) (7.84 g, 6.75 g) over 10 minutes. ml, 51.3 mmol). After the reaction mixture was stirred at room temperature for 20 hours, 5% aqueous KHSO 4 solution (200 ml) was added thereto, and the mixture was stirred vigorously for 15 minutes. The aqueous layer was extracted three times with ethyl acetate. The organic extracts were combined, washed once with saturated NaCl solution, dried over Na 2 SO 4 and concentrated in vacuo. The residue was azeotroped twice with toluene (50 ml). The precipitate formed was suspended in toluene and filtered. The filtrate was concentrated and the azeotropic distillation / filtration process was repeated. The resulting filtrate was evaporated under reduced pressure and then kept under high vacuum provided by a pump for 5 hours. The resulting viscous oil was stirred under argon at room temperature in dry pyridine (50 mL). To this solution was added dicyclohexylcarbodiimide (DCC) (4.65 g, 22.6 mmol) in one portion, followed by the addition of methanol (1.31 g, 1.67 mL, 41.0 mmol). After stirring at room temperature for 20 hours, the reaction mixture was filtered through a pad of diatomaceous earth formed on glass sinter. The diatomaceous earth was washed with ethyl acetate, and the combined filtrates were evaporated under reduced pressure. The residue was redissolved in ethyl acetate and washed twice with 5% aqueous KHSO 4 and once with saturated NaCl solution. The organic extraction solution was dried over Na 2 SO 4 and filtered, the filtrate was concentrated and azeotroped twice with toluene, and the residue was suspended in toluene and filtered. The resulting filtrate was reconcentrated, azeotroped and filtered, and the filtrate was evaporated under reduced pressure and placed under high vacuum for 6 hours to give the phosphonate monoester as a clear viscous oil 64,849,493 (10.2 g, yield> 100%). TLC (silica gel; nPrOH-NH 4 OH-H 2 O, 7: 2: 1): Rf = 0.50. The phosphonate monoester (1.21 g) was kept under high vacuum on a pump for 4 hours to give 1.16 g (2.57 mmol) of ester, which was dissolved in dry ethyl ether (10 mL) and dicyclohexylamine (0.481 g, 0.528 mL, dropwise) was added dropwise. 2.65 mmol). The resulting homogeneous solution was allowed to stand at room temperature for 7 hours, resulting in considerable crystal formation.

The mixture was kept at -20 ° C for 16 hours, after which it was warmed to room temperature and filtered. The crystals were washed with cold dry ethyl ether and then dried under H 2 under high vacuum for 18 hours. The crystals were then kept under high pressure under 45 ° C for 4 hours to give 1.25 g (1.98 mmol, 77% yield) of the title dicyclohexylamine salt as a white powder; mp. 155-156 ° C. TLC (silica gel; MeOH-CH 2 Cl 2, 20:80): Rf = 0.57.

1 H-NMR (270 MHz, CDCl 3): δ 7.71 - 7.65 (m, 4H) 7.40 - 7.32 (m, 6H) 4.02 (m, 1H) 3.52 (s, 3H ) 3.28 and 3.22 (m, 1H) δ 3.11 (d, 3H J = 11 Hz) 2.77 - 2.64 (m, 2H) 2.62 - 2.56 (m, 1H) 1.92 to 1.08 (number of m, 22H) 1.00 (s, 9H) Mass spectrum (FAB): 632 (M + H) + IR (KBr): 3466 to 3457 (broad), 3046, 3016, 2997, 2937, 2858, 2836, 2798, 2721, 2704, 2633, 2533, 2447, 1736, 1449, 1435, 1426, 1379, 1243, 1231, 1191, 1107, 1074, 1061, 1051 and 820 cm-1 Elemental analysis (c22H3] ° 6psi-CH2N2N): calculated: 64.64; H, 8.61? N, 2.22 Found: C, 64.51; H, 8.49; N, 2.18 I. i

Claims (5)

1. A process for the preparation of therapeutically active compounds of the formula O in rp-ch2-ch-ch2-co2rx X EH (CH2) n (I) z in which R is OH, lower alkoxy or lower alkyl, X is -O- or -NH-, n is 1, R 1 X 2 vvv / R 2 Z is the group R 2a R 2a in which R 1, R 2 and R 2a may be the same or different and are each independently H, halogen, lower alkyl, halo- gene alkyl, phenyl, substituted phenyl or Ory in which R 1 is H, alkanoyl, benzoyl, phenyl, halophenyl, phenylsubstituted lower alkyl, lower alkyl, sinamyl, haloalkyl, allyl, cycloalkylsubstituted lower alkyl, adamantylsubstituted or substituted phenyl) substituted lower alkyl, and R * is hydrogen or lower alkyl, and for the preparation of salts thereof, characterized in that a silyl ether of the formula R II-P-CH (VII) (CH 2) Si-C (CH 3) 3 35. C6H5 C6H5 70 89493 wherein R 1 is lower alkyl or lower alkoxy, a silyl ether group is cleaved to form an ester of the formula 0 II R C-P-CH 2 -CH-CH 2 -CO 2 R x X O H (VIII) (CH 2) n Z (R * is alkyl) and the ester is converted, if desired, to a corresponding salt of formula O11 R-P-CH 2 -CH-CH 2 -CO 2 alkali metal X OH (CH 2) n (VIHA) Z 20. which R is lower alkyl or lower alkoxy, to an acid of the formula 0 to R-P-CH 2 -CH-CH 2 -CO 2 H 25. OH (CH 2) n (VII IB) in Z to a corresponding salt of the formula 0 is the same as in 10-P-CH 2 -CH-CH 2 -CO 2 alkali metal X OH in (CH 2) n (VIIIC) Z to 71 89493 or to an acid of the formula 0 II ho-p-ch 2 -ch-ch 2 -CO 2 5. OH (VIIID) (CH2) n z Process according to claim 1, characterized in that X is -O- and R is alkoxy. 3. A process according to claim 1, characterized in that X is -NH- and R is alkoxy. 4. A process according to claim 1, characterized in that the compound is prepared is S-4 - [[[4'-Fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] methoxy] methoxyphosphinyl] -3-hydroxybutanoic acid, methyl ester or monolithium salt, S -4 - [[[4'-fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] methoxy] hydroxyphosphinyl] -3-hydroxybutanoic acid, dilithium salt, (3S) - 4 - [[[4'-fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] -methoxy] methylphosphinyl] -3-hydroxybutanoic acid, monolithium salt, (S) -4- [ [[2,4-dichloro-6- [(4-fluorophenyl) methoxy] phenyl] methoxy] -methoxyphosphinyl] -3-hydroxybutanoic acid, monolithium salt, (3S) -4 - [[[2,4-dichloro-6- [ (4-fluorophenyl) methoxy] phenyl] methoxy] hydroxyphosphinyl] -3-hydroxybutanoic acid, dilithium salt, (3S) -4 - [[[2,4-dichloro-6 - [(4-fluorophenyl) methoxy] phenyl] methoxy] methylphosphinyl] -3-hydroxybutanoic acid, methyl ester, or (S) -4 - [[[[4'-fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl) -2-yl] -methyl] amino] methoxyphosphinyl] -3-hydroxybutanoic acid, monolithium salt. 5. Intermediate, characterized in that it has the structure 72 8 9 493 o Ra-P-CH 2 -CH-CH 2 -CO 2 alkyl Oalkyl OSi-C (CH 3), alkoxy or OH, and which structure contains all stereoisomers of the intermediate. IN: