DD270714A5 - PROCESS FOR THE PREPARATION OF PHOSPHOROUS HMG-COA REDUCTASE INHIBITORS - Google Patents

Abstract

The invention relates to a process for the preparation of phosphorus-containing compounds which inhibit the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase and are thus suitable as hypocholesterolemic agents. The compounds prepared according to the invention have the formula including their salts, in which R is a hydroxyl group, a lower alkoxy or lower alkyl radical; Rx is a hydrogen atom or an alkyl radical; X is an oxygen atom or the group NH, n is 1 or 2, and Z is a hydrophobic anchor. formula

Description

Title of the invention

Process for the preparation of phosphorus-containing HMG-CoA reductase inhibitors

30

35

Field of application of the invention

The application of the present invention takes place in the area of _r .Medizin in inhibiting the Aktvität j S-hydroxy-S-methylglutaryl-coenzyme A reductase and thus inhibiting the biosynthesis of cholesterol.

Characteristic of the known state of the art

F.M. Singer et al., Proc. Soc. Exper. Biol. Med., Vol. 102 (195), p. 370 and F.H. Hulcher, Arch. Biochem. Biophys., Vol. 146 (1971), p. 422, report that certain mevalonate derivatives inhibit the biosynthesis of cholesterol.

Endo and Mitarb, U.S. Patents 4,049,495, 4,137,322 and 3,983,140, report a fermentation product which effects the inhibition of cholesterol biosynthesis. This product is referred to as compactin and Brown and Mitarb report in J. Chem. Soc. Perkin I. (1976), p. 1165, shows that it has a complex mevalonolactone structure.

Tn GB-A-1,586,152 is a group of synthetic compounds of the formula

t ' · ""'''" \ ·· \ - '-'''"" \.. '.' * - '. · ·', '"".

^ '* ^{:; r ;} '''' 2707 M 'ν

in which E denotes a direct bond, an alkylene bridge having 1 to 3 C atoms or a vinylene bridge and the various radicals R signify a multiplicity of substituents. The activity reported in the GB patent is less than 1 % of that of COmpactin.

In US-A-4,375,475 hypocholesterolemic and hypolipidemic compounds of the structure '' · '

H 20

E is a direct bond or one of the groups - ^CH 2 "" - ^CH ₂ -CH ₂ - ,, -CH ₂ -CH ₂ -CH ₂ - or -CH = CH- "in which A is a hydrogen atom or a methyl group. R ₁ , R ₂ and R ₃ each represents a hydrogen or halogen atom, a C ₁₄ alkyl, Cj_ ₄ haloalkyl, phenyl, halogen substituted phenyl, C 1-4 alkoxy, C ₂ "alkanoyloxy C 1 -C 4 -alkyl or C 1 -C 4 -haloalkyl radical or the radical OR ₄ , in which R 1 is a hydrogen atom, a C 1-6 -alkanoyl-, benzoyl-,

4 Z - o.

Phenyl, halophenyl, phenyl-C.galkyl, C ^ g

Cinnamyl, C.-haloalkyl, allyl, cycloalkyl-C. 'alkyl, adamantyl C ₁ _ ₃ alkyl or a substituted phenyl-C. , -alkyl radical, where the substituents are each halogen atoms, Ο.,.-Alkoxy-, C.. -alkyl or C.. -haloalkyl radicals. Furthermore, the corresponding dihydroxy acids resulting from the hydrolytic opening of the lactone ring are described. The US patent further describes the pharmaceutically acceptable salts of these acids and the C., -alkyl- and the phenyl-, dimethylamino- or acetylamino-substituted C _1-3 -alkyl esters of the dihydroxy acids. All of these compounds are enantiomers having a 4 R configuration in the tetrahydropyran unit of the tear racemate of the above formula.

WO 84/02131 (PCT / EP83 / 00308) describes heterocyclic analogues of mevalolactone and derivatives thereof of the structure

X-Z

in the one of the radicals R and R is a radical of the formula

-QC

and the other is a primary or secondary C ₁ _g alkyl, C ₃ g-cycloalkyl or phenyl- (CH _{2) m} group, wherein R ₄ is a hydrogen atom, a C ^ alkyl, C _{1- 4} -alkoxy (with the exception of the tert-butoxy radical), trifluoromethyl, fluorine, chlorine, phenoxy or benzyloxy radical,

2 7E7 14

R _{5 is} a hydrogen atom, a C 1-4 alkyl, C 1-6 alkoxy,. Trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy,

R ₁ . a hydrogen atom, a C,., - alkyl, C ₁ - alkoxy, fluorine or chlorine radical, Rnd

m is 1, 2 or 3, ·

with the provisos that both R _g and R ₅ must be hydrogen when R 1 is hydrogen, R ₅ must be hydrogen when R _{5 is} hydrogen, not more than one of R and R _{5 is} one Not more than one of the radicals R ₄ and R ₅ denote a phenoxy group and not more than one of the radicals R. and, R 'signify a benzyloxy group,

V Ro represents a hydrogen atom, a C, ₄ ~ alkyl, C ₃ g -cycloalkyl, C ₁ -alkoxy (excluding the tert-butoxy group), trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy group, R ₁ represents a hydrogen atom, a C ₁ , -C 4 alkyl, C ₁ -alkoxy, trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy group, with the provisos that R ₃ must be a hydrogen atom, when R_ is a hydrogen atom, not more than one of R ₂ and R _{3 is} a trifluoromethyl group, not more than one of R ₂ and R _{3 is} a phenoxy group, and not more than one of R and R, a benzyloxy group represents,

, X is one of the radicals "(CH ^ - or -CH = CH- (n = 0, 1, 2 or 3), Z is a radical of the formula

^R 6 5 4 3 2 1

-CH-CH ₂ -C-CH ₂ -COOH II OH OH

means _;

wherein R _{fi is} a hydrogen atom or a C ₁ - alkyl radical in the form of the free acid or in the form of a physiologically hydrolysable. and a compatible ester or an o-lactone thereof or

!, Ho-L ¹

A -5-

in the form of a salt thereof.

10 15

GB-A-2-162-179 describes naphthyl analogs of mevalolactone which are useful as inhibitors of cholesterol biosynthesis and the structure

in which R _{1 is} a C ₁ --Alkylrest, Z represents a radical of the formulas Z ₁ or Z ":

-CHCH ₂ CHCh ^ COOR ₇ OH OH

_k 0H "'K

(Z ₁ )

(Z ₂ )

and R represents a hydrogen atom, a hydrolyzable ester group or a cation.

EP-A-164-698 describes the preparation of lactones which are useful as antihypercholesterolemic agents

30 35

Treatment of an amide with an organic sulfonyl halide of the formula R SO ₂ X, after which the protective group Pr is removed. The reaction proceeds according to the following reaction

equation

R ³ R

R ⁵ SO ₂ X

I. 5so-

':' ; Ι '* ^: -

• f; α.ι

where X is a halogen / itom,

Pr represents a carbinol protecting group, R represents a hydrogen atom or a methyl group,

3 4 '

R and R each represent a hydrogen atom, a C, ~~ alkyl ~ or phenyl-C ^ g-alkyl radical, wherein the phenyl group is optionally substituted by a C _1-3 alkyl, C ^ j alkoxy or

a halogen atom may be substituted,

2 R is a radical of the formulas (A) or (B)

(A)

Q is a radical of the formula

Γ ι

CH

or

R ⁶ is -CH

represents,

R is a hydrogen atom or a hydroxyl group, R is a hydrogen atom or a methyl group, a, b, c and d are optionally double bonds, R is a phenyl or benzyloxy group, where the ring is in each case optionally substituted by a C ₁ , -alkyl-

radical or a halogen atom is substituted,

8 'R and R each represent a C., -alkyl radical or a halogen atom

represent,.

R is a C. alkyl, phenyl or a mono- or di

alkyDphenyl radical represents. ,

07 1 4

-7-

In DE-A-35 25 256 are naphthyl analogues of mevalolactones of the structure

le

• GHCH-jCHCH-CO-R OH OH

in which R represents an alkyl radical, the radical Z represents one of the radicals Q or Q and R represents a hydrogen atom or a hydrolyzable ester radical which are suitable as inhibitors of cholesterol biosynthesis and for the treatment of atherosclerosis.

In WO-84/02903 are suitable as hypolipoproteinemische agents mevalolactone analogues of the formula

X-Z

in which the two radicals Rc together form a radical of the formulas

8 7 6 5 C = C-C = C

or -

35 mean, where

R _{2 is} a hydrogen atom, a C., -alkyl, C _1-4 -alkoxy- (

2 70 7 f4 "

taken from tert-butoxy), trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy, R- ^e; i- ⁿ hydrogen atom, a C ₁ --Alkyl-, C. --Alkoxy-, trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy

* '5, with the provisos that no more than one of R- and R- represents a trifluoromethyl group, not more than one of R- and R- represents a phenoxy group and not more than one of R- and R- _{3 represents} a benzyloxy group,

R _{1 is} a hydrogen atom, a C._galkyl, fluoro, chloro; or benzyloxy radical,

R- is a hydrogen atom, a C ₁ , -alkyl-, - C. ₄ alkoxy (excluding tert-butoxy), trifluoromethyl, fluoro, chloro, phen

; oxy or benzyloxy radical,

R _{g represents} a hydrogen atom, a C ₁ - alkyl, C ₁ , - alkoxy, trifluoromethyl, fluoro, chloro, phenoxy or benzyloxy radical,. , j

R _{5 represents} a hydrogen atom, a C ₁ alkyl or C ₁ alkoxy radical, a fluorine or chlorine atom, with the provisos that not more than one of R ₁ and R _{5 is} a trifluoromethyl group, not more than one R and Rg represent a phenoxy group and not more than one of R, and R _{5 is} a benzyloxy group, X is a radical of the formulas H

where η is 0, 1, 2 or 3 and both q are 0 or one is 0 and the other is 1, Z is a radical of the formula

«6 5 4 3 | 2!

-CH-CH-CH- ₀ -C -COOH II I ^Δ I * OH OH

707/4

-9-

represents where Rg represents a hydrogen atom or a C. --Alkylrest, with the general proviso that the -XZ and are the radical R ₄ phenyl group bearing ortho to each other.

The compounds are described in the form of the free acid or in the form of a physiologically hydrolyzable and compatible ester or a δ-lactone thereof or in the form of a salt.

In EP-A-127.848 derivatives are ylalkansäuren of _{3-hydroxy-5-thia-ar} 0ü- the structural formula

HO ^

SO)

described in the Z a remainder of the formulas

means that η is 0, 1 or 2

E is one of the groups -CH ₃ -, -CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH ₂ -, -CH = CH-CH ₂ - or -CH ₂ -CH = CH-, RR ₂ and R _{1 is,} for example, hydrogen, chlorine, bromine or fluorine atoms, C _1. -Alkyl, phenyl or substituted phenyl radicals, or the radical -OR ₇ , in which R _{7 is,} for example, a hydrogen atom, a C ₂ _g-alkanoyl-, Benzoyl, phenyl, substituted

phenyl,

, Cinnamyl, C 1-4 haloalkyl, allyl

L > ·

70 7 f 4 '

- 10 -

IyI-, cycloalkyl-C. represents --alkyl, "adamantyl-Cj ^ -alkyl or Phe -, - alkyl radical,

λ ε a R, R and R are hydrogen, chlorine, bromine or fluorine atoms

a or / C ₁ --Alkylrest, and

X e.g. represents a hydrogen atom, a C 1 - alkyl group, a cation derived from an alkali metal atom, or an ammonium ion.

These compounds have the cholesterol lowering effect due to their ability to inhibit 3-hydroxy-3

methylglutaryl-coenzyme A (HMG-CoA) reductase and fungicidal efficacy.

In FR-A-2,596,393 3-carboxy-2-hydroxypropanphosphon- acid derivatives are described ¹ ^ and salts thereof, which are useful as hypolipidemic agents and the formula

h , V ^0R 3

R ₁ OOC-CH ₀ -C-CH ₀ -P ^ 20 ^{0H 0R} 4

in which R ₁ and R _{2 are} hydrogen atoms, lower alkyl radicals or optionally substituted Aralkylrestebedeuten,

R ₃ and R _{4 represent} hydrogen atoms, lower alkyl radicals or optionally substituted aryl or aralkyl radicals.

These compounds are said to have a greater reduction in cholesterol, glyceride and phospholipid levels than meglutol

yield

 30

- 11 -

In EP-A-142,146 mevinolinartige compounds of the formula

HO _ ο ·

,1

in which R is, for example, a hydrogen atom or a C.A.Alkylrest, E is one of the groups -CK ₂ CH_-, -CH = CH- or - (CH-) -, and Z "I) is a radical of the formula

9

in which X represents -O- or -NR -, where R represents a hydrogen atom or a C ₁ , -alkyl radical,

7 R is a C_ "-alkyl radical, and

R ^{8 represents} a hydrogen atom or a methyl group, 2) a radical of the formula

2 ·

10

15

270 7 (4

- 12 -

where R, R and R independently of one another are, for example, hydrogen or halogen atoms or C 1, -alkyl radicals,

3) a radical of the formula

1 4, where η is 0 to 2 and R is a halogen atom or

represents a C.-alkyl radical, or 4) a radical of the formula

20

represents.

These compounds are HMG-CoA reductase inhibitors. 25

( Object of the invention

The aim of the present invention is to provide novel phosphorus-containing compounds which inhibit the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) and are thus suitable as hypocholesterolemic agents.

Explanation of the essence of the invention

The invention has for its object to provide new phorphorhaltige HMG-CoA reductase inhibitors available.

The invention thus provides a process for the preparation of compounds of general formula I.

i '';' 1i. f W ι - U "t 'JU"

, , , ..- .'i · LV, 0 0 0 ¹ ^ ·

: H ₂ CH-CH ₂ -CO ₂ alkyl

Si-C (CHa) ₃ ²⁵ ₆ ZC 'h _s ^X C ₆ H ₅

in which R ^c is a lower alkyl or lower alkoxy radical, a Silylätherspaltung subjects to an ester of

formula

R ^C -P -CH ₂ -CH-CH ₂ -CO ₂ R ^X X δΗ

2,707 f4

- I2b -

in which R ^{x is} an alkyl radical, and optionally the ester in the corresponding salt of the formula

RP-CH ₂ -CH-CH ₂ -CO ₂ Alkali metal X OH (CH ₂ ) _n Z

in which R is a lower alkyl or lower alkoxy, or in the acid of the formula

U RP-CH ₂ -CH-CH ₂ -CO ₂ H

^v X OH

l

(CH ₂ ) _n

or in the corresponding salt of the formula

Il

Alkali metal O-P-CHj, -CH-CH ₂ - «^ Alkali metal X ÖH

or in the acid of the formula

O HO-P-CH ₂ -CH-CH ₂ -CO ₂ HX O H I

(CH ₂ ) _n

converts

 35

2707

- 13 -

The term "hydrophobic anchor" as used herein refers to a lipophilic group which, when bound by the appropriate linker ("X") to the HMG-like upper side chain of the molecule, binds to a hydrophobic pocket of the enzyme which is not is used for the binding of the substrate HMG CoA, which results in an increased efficiency compared to compounds in which Z is hydrogen _/ results.

In preferred embodiments, the compounds prepared according to the invention have the formula I.

^! 0 H

"ι ν

RP-CH - C-CH, -C0.R ^X 'ι 2 r 2 2

X OH

(CH ₂ ) _n · j

wherein R is a hydroxyl group, a lower alkoxy or lower alkyl group;

R ^{x is} a hydrogen atom or a lower alkyl radical; χ · is an oxygen atom or an -NH group; η is 1 or 2; ζ is a hydrophobic anchor;

, the preferred embodiments also include the salts and the pharmaceutically acceptable salts of these compounds.

The term "salt" refers to basic salts formed with inorganic and organic bases. Such salts include ammonium salts, alkali metal salts such as lithium, sodium and potassium salts (which are preferred), alkaline earth metal salts such as the calcium and magnesium salts, salts with organic bases such as amine-like salts, for example

nc. ,

example, the dicyclohexylamine, benzathine, N-methyl-0-glucamine and hydrabamine salts, salts with amino acids such as arginine or

\ ; 2ί. · · Ν · ΐ · L -, ί /. Cl

- 14 .-

Lysine. , , The non-toxic, pharmaceutically acceptable salts are preferred, although other salts are also useful, for example for isolation or purification of the product.

Examples of hydrophobic anchors according to the invention are (but not limited to)

or

where _<e j ^ dotted lines represent possible double bonds, for example, radicals of the formulas

alkyl

(R ^6a )

alkyl

 2i υ

ίιϋυ- ι ' .GoC.

alkyl

alkyl

alkyl

or

In the above formulas, R ¹ , R ² , R ^2a and R may be the same or different and are independently hydrogen or halogen, lower alkyl, haloalkyl, phenyl or substituted phenyl, or radicals of the formula OR ^, where R ^ represents a hydrogen atom, an alkanoyl, benzoyl, phenyl, halophenyl, phenyl, lower alkyl, lower alkyl, cinnamyl, haloalkyl, allyl, cycloalkyl, lower alkyl, adamantyl, lower alkyl or substituted phenyl lower alkyl radical.

- 16 -

2 7e7 f4

If Z is the rest of the formula

.6

10

alkyl

15

5 5 ', the radicals R and R are identical or different and denote hydrogen atoms, lower alkyl radicals or hydroxyl radicals.

Il

groups,

R ⁶ is lower alkyl-C, for example CH ₃ -CH ₂ -CC-,

or aryl CH ₂ - 'CH ₃ R ⁷

R is a lower alkyl radical, a hydroxy or oxo group or a halogen atom,

q is 0., 1, 2 or 3 and

R represents a hydrogen atom or a lower alkyl radical.

Thus, the compounds of the formula I comprise the following structures:

25

30

0 RP-CH ₀ -QH-CH ₀ -C0 _o R ^x

I Δ - £ ί

0 OH

<f ^H 2> n Z

(IA)

and

35

0 ii

RP-CH ₂ -CH-CH ₂ -CO ₂ R 'OH

NH

(IB)

- 17 -

The terms "lower alkyl" or "alkyl" mean, alone or as part of another group, unbranched or branched hydrocarbon radicals having 1 to 12 carbon atoms in the normal chain, preferably 1 to 7 carbon atoms, such as methyl, ethyl, propyl, la-propyl, butyl, tert-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl -, undecyl and dodecyl group, the various branched-chain isomers thereof, and further such groups having a halogen substituent, such as a fluorine, bromine, chlorine or iodine atom or a trifluoromethyl group! an alkoxy, aryl, alkylaryl, haloaryl, cycloalkyl, alkylcycloalkyl, hydroxy, alkylamino, alkanoylamino, arylcarbonylamino, nitro, cyano, thiol or alkylthio substituent.

The term "cycloalkyl radical", alone or as part of another group, denotes saturated cyclic hydrocarbon radicals having 3 to 12, preferably 3 to 8 carbon atoms.

atoms. These include cyclopropyl, cyclobutyl, cyclopene

tyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, each containing 1 or 2 halogen atoms, 1 or 2 lower alkyl, 1 or 2 lower alkoxy, 1 or 2 hydroxy, 1 or 2 alkylamino, 1 or 2 alkanoylamino, 1 or 2 arylcarbonylamino, 1 or 2 amino, 1 or 2 nitro, (_ 1 or 2 cyano, 1 or 2 thiol and / or 1 or 2 alkylthio may be substituted.

The term "aryl radical" denotes monocyclic or bicyc-30

aromatic, radicals having 6 to 10 carbon atoms in the ring moiety, such as the phenyl, naphthyl, substituted phenyl or substituted naphthyl, wherein the phenyl or naphthyl group 1, 2 or 3 lower alkyl radicals, halogen atoms

(Cl, Br or F), 1, 2 or 3 lower alkoxy, 1, 2 or 3 35

Hydroxyl groups, 1, 2 or 3 phenyl groups, 1, 2 or 3 alkanoyloxy radicals, 1, 2 or 3 benzyloxy groups, 1, 2 or 3 halo

n ijüir l --.-! u ;; u ^

2707 f 4

- 18 -

logenalkylreste, 1, 2 or 3 halophenyl, 1, 2 or 3 allyl groups, 1, 2 or 3 Cycloalkylalkylreste, 1, 2 or 3 Adamantylalkylreste, 1, 2 or 3 Alkylaminoreste, 1, 2 or 3 alkanoylamino, 1, 2 or 3 Arylcarbonylaminoreste , 1,2 or 3 amino groups, 1, 2 or 3 nitro groups, 1, 2 or 3 cyano groups, 1, 2 or 3 thiol groups and / or 1, 2 or 3 alkylthio radicals, wherein the aryl radical preferably contains three substituents.

The terms "aralkyl radical", "arylalkyl radical" or "aryl lower alkyl radical", by themselves or as part of another group, denote lower alkyl radicals as described above having an aryl substituent, such as the benzyl group.

The terms "lower alkoxy", "alkoxy", "aryloxy", or "aralkoxy" refer, alone or as part of another group, to the lower alkyl, alkyl, aralkyl or aryl groups described above attached to

20 oxygen atom.

The terms "lower alkylthio radical", "alkylthio radical", "arylthio radical" or "aralkylthio radical", by themselves or as part of another group, signify one of the lower alkyl, alkyl, aralkyl or aryl radicals described above attached to a sulfur atom.

The terms "lower alkylamino", "alkylamino", "arylamino", "arylalkylamino", alone or as part of another group, refer to one of the loweralkyl, alkyl, aryl or arylalkyl radicals described above attached to a nitrogen atom.

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

The term "halogen atom" denotes chlorine, bromine, fluorine and iodine atoms as well as trifluoromethyl groups, where the chlorine

and fluorine atom are preferred. 5

Preferred compounds of the formula I have the following structure:

\ i? / ^CH 2 \ ^H / ^H 2 \ _χ PC / CO ₂ R ^X

CH ₂ OH

(Ii)

wherein R is hydroxyl or OLi; R ^{x is} lithium or hydrogen; X is oxygen or -NH- and

Z is the formula .λ_ ..

R ¹ IR ²

· ο

where R is a phenyl group comprising an alkyl and / or halo-substituent; or R ^{1 is} a benzyloxy group comprising a haloio substituent; R ² and R ^{2a are the} same and a halogen atom or a lower

Q _{0 is} alkyl

20 Z can preferably also the formula

O

1

in which R and R are as defined immediately above with respect to the compound of formula II, or Z may be the formula

have, in which R is a Wasserstoffatan, an IfethyI or a hydroxyl group, and R is a radical of the formula

.CH,

O Il

or a (substituted) phenyl'JiBthylgruppe

CH,

wherein R is a hydrogen atom or an ethyl group.

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

I R ^a

OSi-piai,), R - P (oalkyl) ₂ ^ O

' ^{3 3} III. ν. R ^a -P-CH-CH-CH ^ -CO-alkyl

6 5 Tv y '=.

I-CH ₂ -C-CH ₂ -CO ₂ alkyl Arbuzov reaction Oalkyl QSx-C (CH ₃ J ₃

(R ^a = bodice alkyl or _r "\"

Lower alkoxy) 6 ⁿ 5 ^U 6 ⁿ 5

IV

1. (ClLjK-SiBr, CH-Cl ₉ u

^JJ R ^a -P-CH _o -CH-CH _o -CO, alkyl

XV ^. H ^ U

^ OH _y ,

Phosphorus ester: / Si-C (CH,), κ,

cleavage ^

-6 ^H 5 ^C 6 ^H 5

a 'a

VA. R = lower alkyl when R was lower alkyl

VB. - R ^a = OH,. when R ^{a was} lower alkoxy

^Rb OH-DCC

^OH) (R ^b = lower alkyi)

pyridine

-Veresterung

Il

R ^D OP-CH ₂ -CH-CH ₂ CO ₂ aikyl OH

Si-C (CH ₃ ) 3

^C 6 ^H 5 ^C 6 ^H 5

VI

VI 1) or 2) VA.

(COCl) ₂ ; CH ₂ Cl ₂ (acid chloride) ^Z "< ^CH 2 ⁾ _n - ^XH ( coupling reaction)

(C ₃ H ₅ J ₃ N, DMAP

1) In-C ₄ H ₉ J ₄ NF, CII ₃ COOH, THF (silyl

Ether splitting

2) oh ", - dioxane (hydrolysis)

R -f-CH ₂ -gH-CH ₂ CO ₂ alkyl

(CIi ₇ ) Si-C

VII

Lower alkyl or lower alkoxy)

L-P-I

' ^2W 2

OH

2707 f4

From the reaction sequence given above, it can be seen that compounds of formula I can be prepared by subjecting the iodide A to an Arbuzov reaction by reacting the iodide A

^C 6 ^H 5 O-Si-C (CH ₃ ) ₃

I ^C 6 ^H 5 (A)

I-CH ₂ ^ C-CH ₂ -CO ₂ alkyl

H ^-

 ,

and the phosphonite / phosphite I-II heated

R ^a -P (Oalkyl) ₂ 'III)

wherein R ^{a is} a lower alkyl or lower alkoxy, using standard Arbuzov conditions and procedures to obtain the phosphinate / phosphonate IV.

RP CH ₂ CH-CH ₂ -CO ₂ alkyl

Oalkyl OSi-C (CH ₃ ) ₃ ^(IV)

^C C ^H C ODOO

tias Phosphorin / wiosnhona.t iv is a novel compound and as such forms part of the invention

 25

The phosphinate / phosphonate IV is then subjected to phosphorus ester cleavage by dissolving a solution of compound IV in the presence of an inert organic solvent such as methylene chloride and under an inert gas such as argon.

subsequently treated with bis (trimethylsilyl) trifluoroacetamide (BSTFA) and trimethylsilyl bromide. The phosphinic acid VA is obtained

- 25. -

Lower alkyl-If-CH ₂ -C ₅ H-CH ₃ -CO ₂ alkyl (VA)

OH 0. S

^C 6 ^H 5 ^C 6 ^H S

in which the lower alkyl radical corresponds to the radical R ^a of compound IV, or the phosphonic acid VB · o

HO-P-CH ₀ -CH-CH ₀ -CO ₀ alkyl OH 0,

Si-C (CH ₃ ) ₃

^C 6 ^H 5 ^C 6 ^H 5

in the lower alkoxy group. the radical R of compound IV corresponds. * '

Compounds VA and VB are novel intermediates and, as such, a part of the invention.

The resulting phosphonic acid VB is esterified by reacting VB in the presence of dry pyridine with an alcohol VC

R ^b OH (where R ^{b is} a lower alkyl radical) (VC)

ν Dicyclohexylcarbodiimid.behandelt and the reaction mixture obtained and under an inert gas such as argon is stirred · Man. receives the Phosphonmonoalkylester VI

R ^{b is} Oi-CH ₂ -QH-CH ₂ CO ₂ alkyl

OH 0

^ Si-C (CH ₃ ) ₃

C ^ He C _fi Hc 6 5 6

1 1 \ Wl · - / I ^1. , 6.;>. Ä;,; '

Ester VX or phosphinic acid VA is then dissolved in an inert organic solvent / such as methylene chloride, benzene or tetrahydrofuran (THP), treated with trimethylsilyldiäthylamin and stirred under an inert gas such as argon; the mixture is evaporated and then dissolved in methylene chloride (or other suitable inert organic solvent). The resulting solution is cooled to a temperature in the range of about ⁰ ° C to about 25 ^° C, treated with oxalyl chloride and then evaporated. Man. receives crude phosphonochloridate. The resulting phosphonochloridate is dissolved in an inert organic solvent such as methylene chloride, benzene oil, pyridine or THP; the solution is cooled to a temperature in the range of about -2O ⁰ C to about O ⁰ C, with

Z- (CH ₂ ) _n -XH (B)

treated, wherein the molar ratio of ester VI or: phosphinic VA: B in the range of 0.5: 1 to 3: 1 and preferably from 1: 1 to 2: 1, then with triethylamine go ^and catalytic 4-dimethylaminopyridine (DMAP ). Man receives the addition product VII

j! CH ₂ : CH-CH ₂ -CO ₂ alkyl

X «?H ₂ ) _n ^ iC (CH ₃ ) 3 (VII)

Z is ^C 6 ^H 5 ^C 6 ^H 5 in which R ^{c is} a lower alkyl or lower alkoxy radical.

Compound VII is subjected to silyl ether cleavage by treating Compound VII in the presence of an inert organic solvent such as tetrahydrofuran with glacial acetic acid and tetrabutylammonium fluoride. The ester VIII is obtained

11 -JiJJo- b'i.tK · G.

4. 1.1 -JiJ

-27 -

If -CH ₂ -QH-CH ₂ -XOH

-5 Z

(R ^x = alkyl)

The ester VIII may then be hydrolyzed to the corresponding alkali metal salt or acid, ie a compound in which R ^{x is} an alkali metal or hydrogen atom. By

Treatment of ester VIII at 25 ⁰ C with a strong base such as lithium hydroxide in the presence of dioxane, tetrahydrofuran or other inert organic solvent and under an inert gas such as argon, wherein a molar ratio of base to ester VIII in the range of about 1: 1 to about 1.1: 1, the corresponding alkali metal salt VIIIA '''

'20 R-P-CH 1 -CH-CH-j-CO, alkali metal (VIIIA)

l 2 s ί & X OH

«JH ₂ ) _n

in which _R i is _lower alkyl or lower alkoxy.

Compound VIIIA can then be treated with a strong acid such as hydrochloric acid to give the corresponding acid

VIIIB 30

CH "-CH-CH ₀ -CO ₀ H (VIIIB)

X OH

<? ^H 2> n Z

ι s ι

2707 t 4 '

- 28 -

The ester VI7.I having the structure in which R is lower alkoxy can be converted to the corresponding di-alkali metal salt. By treatment of ester VIII with a strong base at from 50 to 6O ⁰ C, with a molar ratio of base to ester VIII in the range of about 2: 1 to about 4: 1 is selected, to give the corresponding dialkali metal salt VIIIC

.0

Alkali metal.0-P-CH ₀ -CH-CH ₀ -CO ₀ .Alkalimetall (VIIIC) X ÖH

«T ^H 2> n

The di- ^A lkalimetallsalz VIIIC can be converted by treatment with a strong acid such as hydrochloric acid to the corresponding acid VIIID °

O HO-P-CH ₂ -CH-CH ₂ -CO ₂ H

X ÖH

(CH ₂ ) _n

in which the radical R is a hydroxyl group. The iodide starting material A can be prepared by dissolving bromide C

C? ^H

^v Br-CH ₂ -CH-CH ₂ CO ₂ alkyl (C)

(prepared according to the method described in Tetrahedron Lett. 26 · .Ί985) '2951), the .in a solution of .. <. Dimethylformamide (DMF) is dissolved with imidazole and 4-dimethylaminopyridine. The resulting solution is treated with tert-butyldiphenylsilyl chloride under an inert gas such as argon. The silyl ether D is obtained

' - 29 -

OSi-C (CH ₃ ) 3 ^(D) Br-CH ₂ -CH-CH ₂ CO ₂ alkyl

Siyl ether D dissolved in an inert organic solvent such as methyl ethyl ketone or DMF is treated with sodium iodide under an inert gas such as argon to give the iodide A.

The starting compound B

Z- (CH ₉ ) -XH (B)

^{Z n}

can be prepared as described below depending on the definitions of Z and X.

Thus, compounds of the formula B in which Z is the formula 20

25

and X is an oxygen atom, compounds of structure B1.

30

> - < ^CH 2> n- ^0H

Compounds of structure B1 can be produced by reacting the aldehyde E

2707f4 '

- ³⁰ "

R ¹

mi ": a reducing agent, such as lithium aluminum hydride or sodium borohydride, compounds of formula B wherein Z is the formula

R ¹

and X is a nitrogen atom, compounds are the _ '

Structure B.

R. /

R ^2a -H ^ 0V- (CH ₂ ,) _n

Compounds of structure B can be prepared by oxidation of the dehydrogen E, by treating Ef dissolved in acetone, for example, with Jones reagent. * ^The acid F is obtained

R ¹

R ^2a - / O V - (CH ₂ J _n-1 -COOH (F)

^ ²

2 70 7

- 31 -

which is treated in suspension in methylene chloride with oxalyl chloride. Man. receives 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. An amide of structure G is obtained

O Il

2 years

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

25

Starting compounds of the formula B in which Z is the formula H

alkyl

(H)

30 35

and X is oxygen or -NH-, are compounds of structure H ¹

2707 i4

- 32 -

Compounds of structure H in which X is oxygen are known from Hethcock et al. J. Org. Chem. 50 (1985), 1190. Compounds of formula H ¹ wherein X is -NH- may be prepared by the r. Reductive amination of J

ίο

^ O CKO

AMV

alkyl

(R ^6a ).

(J)

15

(prepared as known from Hethcock et al., supra) may be prepared by treating J with ammonium acetate and sodium cyanoborohydride in the presence of an alcoholic solvent, such as methanol.

20 25 30

Starting compounds of the formula B in which Z is the formula

and X is oxygen are compounds of structure M.

(M)

(CH ₂ J _n -OH

These compounds are known from WO-A 8402-903 and GB-A 2,162,179.

2 'Aiii' .-

- 33 -

Starting compounds of the formula B in which Z is the formula

^has and _: X is -NH-, structure N connections are

, 2a

(N)

Oj O-HCH ₂ ) _n -NH ₂

Compounds of structure N can be prepared by reductive amination of the aldehyde 0

(O)

are prepared by treating the aldehyde O with ammonium acetate and sodium cyanoborohydride in the presence of an alcoholic solvent, such as methanol.

The compounds can be prepared as racemic mixtures and can later be dissolved to obtain the preferred S-isomers. However, the compounds of the invention may also be prepared directly in the form of their S-isomers as described herein and below in the Examples.

i ir IM i ^; ^

2 7 Ö 7 f 4

<- 34 -

The compounds of the invention are inhibitors of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase and are thus useful for inhibiting cholesterol biosynthesis as in

shown below. 5

1. Rat liver HMG-GoA reductase

Rat liver HMG-CoA reductase activity is determined by a modified method of Edwards et al., J. Lipid Res. 20 (1979), 40. Rat liver microsomes are used as the source of the enzyme, and enzyme activity is measured by measurement.

14 ing the transformation of <

C-Mevalonic acid determined.

14 conversion of the C-HMG-CoA substrate in

a. Preparation of microsomes

From 2 to 4 cholestyramine-fed, decapitated Spraguej Dawley rats, the liver is removed and stored in phosphate buffer A (sodium phosphate, 0.04 M, pH 7.2, KCl, 0.05 M;

Sucrose 0.1 M; EDTA, 0.03 M; Aprotinin, 500 KI units / ml)

homogenized. The homogenate is centrifuged at 16,000 g for 15 minutes at 4 ^° C. The supernatant is removed and centrifuged a second time under the same conditions. The second 16,000 _× g supernatant is centrifuged at 100,000 _× g for 70 minutes at 4 ^° C. Pelleted microsomes are taken up in a (minimum volume of buffer A (3-5 ml per liver) and homogenized in a glass / glass homogenizer.) Dithiothreitol is added (10 mM) and the preparation aliquoted, rapidly frozen in acetone / dry ice and at -80 ⁰ C

outsourced. The specific activity of the first microsomal preparation is indicated by ° /. ⁶ 8 nmoles of mevalonic acid per mg protein / min.

b. Enzyme test 35

The reductase is tested in 0.25 ml of buffer, which contains the following

t'-J- ') Ί

- 35 -

containing components in the indicated final concentrations contains:

0.04 M Sodium phosphate, pH 7.0 0,05 M KCl 0,10 M sucrose 0.03 M EDTA 0.01 M dithiothreitol 3.5mM NaCl r% dimethyl sulfoxide 50-200 μg microsomal protein 100 μπι ¹⁴ C- [DL] HMG-CoA (0.05 μΰΐ, 30-60 mCi / mmol) 2.7 mM NADPH (nicotinamide adenine dipeptide phosphate).

The reaction mixtures are incubated at 37 ^° C. Under de, n described conditions, the enzyme activity increases linearly to a protein concentration of 300 micrograms of microsomal protein per reaction mixture and linear with respect to the incubation time up to an incubation time of 30 minutes. The standard 20 minute drug incubation time results in 12 to 15 percent conversion of the HMG-CoA substrate to the mevalonic acid product. The [DL-] HMG-CoA substrate is used at a concentration of 100 μπι. This concentration is twice the concentration necessary to saturate the enzyme under the conditions described. NADPH is added at a concentration 2.7 times higher than that needed for the maximum enzyme rate.

Standardized tests for assessing inpatients are carried out according to the following procedure. Microsomal enzyme is incubated at 37 ° C for 15 minutes in the presence of NADPH. The vehicle DMSO is added with or without test compound and the mixture is incubated at 37 ^° C. for a further 15 minutes. The

- 36 -

Enzyme assay is started by addition of C-HMG-CoA substrate. After 20 minutes incubation at 37 ⁰ C is the. Reaction stopped by adding 25 μΐ 33% potassium hydroxide solution. H-Mevalonic acid (0.05 μίί) is added and the reaction mixture allowed to stand at room temperature for 30 minutes. 50μΐ 5N hydrochloric acid is added to form a lactone from the mevalonic acid. As a pH indicator is a

Drops of bromophenol blue added. Lactone formation occurs during a 30 minute incubation at room temperature. The reaction mixtures are centrifuged for 15 minutes at 2800 rpm. The supernatants are added to 2 g of AG 1-X8 anion exchange resin (Biorad, formate form) poured into 0.7 cm (inner diameter) glass columns and eluted with 2.0 ml of water. The first 0.5 ml are discarded and the

iK 14

next 1.5 ml and collected. Tritium or C determination in 10.0 ml Opti-Fluorscintillationsflüssigkeit added. The results are calculated as nmol mevalonic acid produced per j 20 minutes and scaled up to 100% of recovered tritium. The drug effects are expressed as IC ₅₀ values (concentration of the drug that results in a 50% inhibition of enzyme activity), which are derived from the dehydrated diet cut-offs indicated with 95% confidence.

The conversion of lactone-formulated drugs to their sodium salts is accomplished by solubilizing the lactone in DMSO, adding a 10-fold molar excess of a sodium hydroxide 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) by using 1N hydrochloric acid and diluted in the enzyme reaction mixture.

2) Cholesterol synthesis in freshly isolated rat hepatocytes

Compounds that show inhibitory activity on HMG-rCoA reductase are judged for their ability to inhibit incorporation

5 14

of C-acetate in cholesterol in i * .rl3ch isolated rat hepatocyte suspensions. For this purpose, methods are used which were originally described by Capuzzi et al., Lipids, 6 (1971), 602.

a) Isolation of rat hepatocytes

Sprague Dawley rats (180-220 g) are anesthetized with nembutol (50 mg / kg). The abdomen is opened and the first branch of the vena porta tied tightly. Heparin (100-200 units) is injected directly into the abdominal vena cava. A single occlusion becomes distal

becomes ί

the vena porta created and the vena porta between the Ver '

and the first branching vein cannulated. The liver is treated with 20 ml / min pre-warmed (37 ° C) isissue.

enriched buffer A (HBSS without calcium or magnesium but with 0.5 mM EDTA) after the vena cava was separated to drain the effluent. The liver is further soaked in 200 ml of prewarmed Buffer B (HBSSj., With .0.'05 bacterial collagenase) After the liver has been soaked in Buffer B, it is excised and decapsulated in 60 ml of Waymouth's single-cell release medium are isolated at room temperature by a 3 minute low centrifugation at 50 χ G. Pelleted hepatocytes are washed once in Waymouth's medium, counted and tested for viability by the exclusion of trypan blue.

These hepatocyte-enriched cell suspensions typically have a 70 to 90 percent viability.

14 b) C-acetate incorporation into cholesterol

35 (

Hepatocytes are incubated with a cell count of 5x10 cells per 2 ml in incubation medium (IM) [0.02 M Tris-HCl (pH 7.4),

tion 4 '

0.1 M KCl, 3.3 mM sodium citrate, 6.7 mM nicotinamide, 0.23 mM NADP, 1.7 mM glucose-6-phosphate].

Test compounds are usually dissolved in DMSO or DMSO: water (1: 3) and added to (IM). The final concentration of .DMSO in (IM) is> 1% and has no significant influence on cholesterol synthesis.

The incubation is started by adding C-acetate (5B mCi / mmol, 2 μΟΙ / πιΙ) and the cell suspensions (2.0 ml) are incubated in 35 mm tissue culture plates for 2 hours at 37 ° C. After incubation The cell suspensions are transferred to glass centrifuge tubes and centrifuged at room temperature for 3 minutes at 50 g The cell pellets are resuspended, lysed in 1.0 ml of water and placed on ice.

Lipids are essentially as though by Bligh,. ... ι. Dyer, Can. Biochem. and Physiol., 37 (1959) 911. The lower organic phase is removed, dried under a stream of nitrogen and the residue is resuspended in 100 μL chloroform: methanol (2: 1). The entire sample is dropped on silica gel (LK6D) thin-layer plates and developed in hexane ^ ethyl ether: acetic acid (75: 25: 1). The plates are scanned and counted using an automated BioScan scanning system. The radioactive label in the cholesterol peak (RP 0.28) is determined and expressed in total counts per peak and% of the label of total lipid extract. Cholesterol peaks in control cultures usually contain 800 to 1000 cpm and account for 9 to 20% of the label in total lipid extract; the results are similar to those of Capuzzi et al. comparable to a 9 percent mark in cholesterol.

The drug efficacy (percent inhibition of cholesterol synthesis) is determined by measuring the percentage labeling in cholesterol for control and drug-treated cultures.

- 39

2707 (4

is compared. Dose response curves are generated from the average of two or three studies and the results expressed as IC _5Q values with a 95% confidence.

3) Cholesterol synthesis in human dermal fibroblasts

Greater inhibitory activity in liver tissue is a feature of a compound having cholesterol synthesis inhibitory activity. Therefore, these compounds are not only tested for their cholesterol synthesis inhibitory activity in hepatocytes but also in cultured fibroblasts. .

a) Human skin fibroblast cultures 15

Human dermal fibroblasts (passage 7-27) are injected into Eagle's minimal medium (EM) containing 10% fetal calf serum; cultured. For each experiment, stock cultures are trypsinized to divide the cell monolayer, counted and in 35 mm

Tissue culture vessels (5 × 10 ⁵ cells / 2.0 ml) plated. The cultures are incubated for 18 hours at 37 ^° C. in 5% CO., / 95% humidified room air. Cholesterol biosynthetic enzymes are induced by removing the serum-containing medium, washing the cell monolayers and adding 1.0 ml of "(EM) containing 1.0% fatty acid-free bovine serum albumin, and incubating the cultures for a further 24 hours.

14

b) C-acetate incorporation into cholesterol .

induced fibroblast cultures are washed with EMEM ₁₀₀ (Earle's minimal medium). Test compounds are dissolved in DMSO or DMSOtEM (1: 3) (final concentration of DMSO in cell culture less than 1.0%) dissolved, added to the cultures and the cultures are pre-incubated at 37 ° C in 5% CO _2/95% 30 minutes humidified room air. After preincubation with drugs will

1 4 (1 - C] sodium acetate (2.0 μΰΐ / πιΐ, 58 mCi / mmol) was added and

- 40 -

the cultures are incubated for a further 4 hours. After incubation, the culture medium is removed and the cell monolayer (200 μg cell protein per culture) scraped off in 1.0 ml of water. Lipids in the lysed cell suspensions are / are extracted by chloroform: methanol as described for hepatocyte suspensions. The organic phase is dried under nitrogen and the resulting residue in chloroform: methanol (2: 1) (100 μΐ) resuspended and the entire sample on silica gel (LK6D) thin-layer plates was dropped and analyzed as f ^ ^r hepatocytes described.

Inhibition of cholesterol synthesis is determined by comparing the percent labeling in the cholesterol peak of control and drug-treated cultures. The results are expressed as IC ₅₀ values and are derived from average values of the dose response curves of two or more experiments. A 95% confidence in the IC _5Q values is also calculated from the average values of the dose response curves

 20

Another object of the present invention are ¹ to pharmaceutical compositions containing at least one compound of formula I together with a pharmaceutical carrier or diluent. The pharmaceutical compositions can be formulated using conventional solid or liquid carriers or diluents, as well as with pharmaceutical additives suitable for the desired mode of administration. The compounds may be given orally, for example in the form of tablets, capsules, granules or powders. They can also be given parenterally in the form of injectable preparations. Such dosage forms contain from 1 to 2000 mg of active ingredient per dosage for use in the treatment. The dose to be administered depends on the unit dose, the symptoms, age and body weight of the patient.

- 41

The compounds of formula I can be administered in a manner similar to the known compounds proposed for use in inhibiting cholesterol biosynthesis, such as lovastatin, to S. mammals, such as humans, dogs and cats. Thus, the compounds of the invention may be administered in an amount of about 4 to 2000 mg in a single dose or in the form

from divided doses i to 4 times a day, preferably in an amount of 4 to 200 mg in divided doses of 1 to 100 mg and especially 0.5 to 50 mg 2 to 4 times a day. Also, administration in sustained-release preparations is possible.

The following examples illustrate preferred embodiments of the invention. Flash (flash) chronography is performed on either Merck 60 or Whatmann LPS-I silica gel. Counterphase chromatography is performed on CHP-20 MCI gel resin <· (Mitsubishi). "In numbers," inch "(2.54 cm).

EXAMPLES

" ^u example!

(S) -4 - [[[4'-Fluoro-3,3 ', 5-trimethyl- [1,1-biphenyl] -2-yl] methoxy] -methoxyphosphinyl-3-hydroxybutyrate monolithium salt

A. N- (2,4-Pimethylbenzyl .i.den) -benzolamine (US-A-4,375,475, p. 39)

A solution of 6.97 ml (50 mmol) of freshly distilled 2,4-dimethylbenzaldehyde (Aldrich) and 4.56 ml (50 mmol) of distillate

Prepared aniline (Aldrich) in 80.0 ml of dry toluene is refluxed for 3.0 hours under argon in a flask equipped with a Dean-Stark apparatus. The mixture is then cooled and evaporated under reduced pressure to a yellow oil. The crude oil is removed by

distillate (0.5 mm Hg, 160 to 1JO ⁰ C), giving 8.172 g (78.1 %) of the desired benzolimine title compound.

10 15

2707 f4

- 42 -

be obtained as a pale yellow oil, which crystallized on standing to a low melting point solid. TLC (4: 1) Hex-acetone, Rf = O, 67 and 0.77 (geometric isomers), U.V. and I.

CH,

(US-A-4,375,475, p. 39)

20 25

A mixture of 6.0 g (28.7 mmol) of Part B A in 144 ml of glacial acetic acid is treated with 6.44 g (28.7 mmol) of palladium (II) acetate. The clear, red homogeneous solution is refluxed for 1 hour under argon. The cloudy mixture obtained is hot durch--: -1/2 "diatomaceous earth in 900 ml H O filtered The precipitated orange solid is dried abfiltrierr under reduced pressure for 16 hours at 65 ° C PoO _{C. Yield:..} 10, 6 g (85.5%) of the desired palladium complex (title compound) as an orange solid, mp 194 to 196 ° C (recrystallized F. an analytical sample according to the literature: 203 to 205 ⁰ C)..

C. 4'-fluoro-3,3 ', 5-trimethyl- [i, 1'-biphenyl] -2-carboxy

aldehyde

(1) Bro m -Ql-fluoro-3-methylphenyi] -magnesium (US-A-4,375,475, p. 37 and 38)

The Grignarü reagent (title compound of part C (1)) is added to 1.35 g (55 g) by dropwise addition of 22.5 g (60.9 mmol) of 5-bromo-2-fluorotoluene (Fairfield Chemical Co.) with stirring , 4 mmol, 8.0 Xq.) Of magnesium turnings in 70.0 ml of anhydrous

- 43 -

^Et 2 ° * ^ne:> - ^ner rate sufficient to maintain the reaction under reflux produced. The reaction is started in an ultrasound facility. After complete bromide addition, the mixture is stirred for 1 hour at room temperature under argon, boiled for 15 minutes under reflux and then allowed to cool to room temperature.

(2) 4'-fluoro-3,3 ', 5-trimethyl-Q, 1'-biphenyl] -2-

carboxaldehyde

In a second flask, a mixture of 3.0 g (6.92 mmol) of the dipalladium complex of Part B and 14.52 g (55.4 mmol, 8.0 eq.) Of triphenylphosphine in 100 ml of anhydrous benzene Stirred for 30 'at room temperature under argon. Then, freshly prepared and filtered (glass wool plug) Grignard reagent of Part C (1) is added in an amount to this solution by cannula, and the mixture is stirred at room temperature under argon for 1.5 hours. Then, 35 ml of '6, ON HCl are added, the mixture is stirred for a further hour at room temperature and then filtered through kieselguhr (1/2 "bed) The filtrate is extracted with 250 ml of Et ₂ O and the extracts are extracted twice with 100 ml Brine, 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

A solid is purified by flash chromatography on 700 g of silica gel and slotted with hexane followed by hexane-Et-O (91: 5). The product fractions are combined and evaporated. Yield: 1.507 g (89.9 %) of the desired aldehyde title compound as a light yellow solid, mp 72-75 ° C.

^ΰυ (literature: 73 to 75 ° C).

TLC: (* »5: 5) HeX-Et ₂ O, Rf = 0.40, UV and PMA.

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

1b ml of anhydrous diethyl ether cooled to ⁰ ° C. in an ice bath is treated with 259 mg (6.82 mmol, 0.55 eq.) Of LiAlH ₄ and the

2 70 7f 4 '

, - 44 -.

gray suspension is added dropwise within 15 minutes with a solution of 3.0 g (12.4 mmol) of aldehyde of Part C in 15 ml of anhydrous diethyl ether. The mixture is stirred for 30 minutes under argon at room temperature, then cooled again to ⁰ ° C. and the reaction is stopped by dropwise addition of 260 μM water, 260 μL 15% NaOH and 780 μM water drop by drop. The suspension is diluted with ethyl acetate, filtered through anhydrous Na ₃ SO ₄ 'over 1/2 "diatomaceous earth, and the colorless filtrate is evaporated under reduced pressure, yield 2.99 g (98.8%) of white solid.

Trituration of the crude solid with cold hexane and Trocknunq under reduced pressure, 2.467 g (81.6%) of the desired title compound as a white solid, mp 102 103 C ^0. TLC: (9: 1) hexane-ethyl acetate, R = 0.24, UV and PMA.

E. ' (S) -3- [C (1 J -dimethylethyl) -diphenylsilyi] -oxy] -4-: (hydroxynexyblyphosphine) -D-butyric acid methyl ester

(1) Methyl (S) -3-bromo-S-hydroxybutyric acid (1) (a) [R- (R *, R *)] - 2,3,4-trihydroxybutyric acid hydrate

calcium salt

(Carbohydrate Research, Vol. 72 (1979), pp. 301 to 304)

50 g of potassium carbonate are added to a solution of 44.0 g (250 mmol) of D-isoascorbic acid in 625 ml H_O. The solution is cooled in an ice bath at 0 ⁰ C and treated batchwise with 100 ml of 30 % H ₂ O-. The mixture is then stirred for 30 minutes in the oil bath at 30 40 ⁰ C bic. Then 10 g of Darco are added and the black suspension is heated on a steam bath until the 0 ^ evolution ceases. The suspension is filtered through diatomaceous earth and evaporated under reduced pressure at a Badtenperatur of 40 ⁰ C. The residue is taken up in 50 mL H 2 O, heated on a steam bath and treated with CH 2 OH until the solution becomes cloudy. The precipitated gummy solid is filtered off and dried in air. Yield: 30.836 g (75.2 %) of the desired

· '' '

2707 f4

, - 45 -

1 calcium salt as a powdery white solid. TLC (7: 2: 1) 1PrOH-NH ₄ OH-H ₂ O, R f = 0.19, PMA.

(1) (b) [s- (R *, S *)] - 2,4-dibromo-3-hydroxybutyric acid

methylester

(K. Bock et al., Acta Scandinavica (B), Vol. 37 (1983), pp. 341-344)

30 g of calcium salt of part (1) (a) are dissolved in 210 ml of 30 to 32 % HBr in acetic acid and stirred for 24 hours at room temperature. Then 990 ml of methanol are added to the brown solution and they are stirred overnight. The mixture is evaporated to an orange oil, taken up in 75 mL of CH ₃ OH, refluxed for 2.0 hours and then evaporated. The residue is partitioned between 100 ml EtOAc and H ₂ O, the organic phase washed twice with H ₂ O and brine and then dried over anhydrous Na ₃ SO ₄ and evaporated. Yield: 22.83 g (90.5 %) of crude dibromide as a light orange oil

 TLC (1: 1) EtOAc-Hex, Rf = 0.69, UV and PMA.

(1) (c) (S) -4-bromo-3-hydroxybutyric acid methyl ester

(Ref. (1) (b)) "25

An argon-purged solution of 20.80 g (75.4 mmol) of the dibromide and 21.0 g of anhydrous NaOAc in 370 mL of EtOAc and 37 mL of glacial acetic acid is treated with 1.30 g of 5 % Pd / C stirred black suspension under hydrogen with a pressure of ^ow 1 bar. The hydrogen uptake is monitored.

After 2.0 hours, the hydrogen uptake is complete and the mixture is filtered through diatomaceous earth. The filtrate is washed with saturated NaHCO 3 solution and brine

and then over anhydrous MgSO4. dried and too raw

Dibromester evaporated as a brown oil. The crude oil is added with another approach (starting from 3d, Π g dibromide)

^r . : c

2,707 f4

- 46 -

combined and distilled under reduced pressure. Yield; 25.77 g (61.3 %) of the desired bromoester title boron * ·· tion as a colorless oil of bp. 79 to 8O ⁰ C (1.0 mm Hg). TLC (1: 1) EtOAc-Hex, Rf = 0.44, PMA.

analysis For C ₅ H ₉ O ₃ Br: 30 C • 4 H br , 56 calc .: 29 48 4 60 40 , 86 gef. : , 76 50 39

(2) (s) -4-bromo-3 - [[(1, i-dimethylethyldiphenyl)

silyl] oxy] butyric acid methylester

A solution of 4.0 g (20.4 mmol) of bromohydrin from part B (1), 6.94 g (5.0 eq) of imidazole and 12 mg (0.005 eq) of 4-dimethylaminopyridine (4-DMAP) in 40 ml of anhydrous DMF is treated with 5.84 ml (1.1 Xq.) Of tert-butyl-diphenylsilyl chloride

.0

and the homogeneous mixture was stirred under argon at room temperature for 15 hours. The mixture is partitioned between 5 % KHSO ₄ and EtOAc, the organic phase washed with H, O and Koc brine, over anhydrous Na ₃ SO ₄ . dried and evaporated. Yield: 9.32 g (100 %) of crude silyl ether as

colorless viscous oil.

TLC (3: 1) hex-EtoAc, Rf silyl ether = ^ 0.75, U.V. and PMA.

(3) (S) -4-Iodo-3 - [[(1, i-dimethyl-ethyl-diphenyloxy] -butyric acid methyl ester

A solution of 9.32 g (201 mmol) of crude bromide from part E (2) in 60 mL of methyl ethyl ketone (dried over 4 ft sieve) is added 15.06 g (100.5 mmol, 5.0 eq) of sodium iodide treated. The yellow suspension is refluxed for 5 hours under A, rgon. The mixture is cooled, diluted with EtOAc, filtered and the filtrate washed with dilute NaHSO ₃ solution until it is colorless. It is then washed with brine, dried over anhydrous Na ₃ SO 4 and evaporated under reduced pressure to give 10.17 g of yellow oil, ras

QS ι · 7Βϋ6.

- 47 -

crude oil is purified by flash chromatography on 600 g of silica gel and eluted with hexane-CH ₂ Cl ₂ (3: 1). The product fractions are combined and evaporated. Yield: \.

7 ', .91 g (74.2 % overall yield for both steps) of the desired

te. Iodide title compound as a clear, colorless, viscous oil. * TLC (3: 1) Hex-EtOAc, product. Rf = 0.75, U.V. and PMA. **

(Note: the iodide product gives a spot in the same place as the bromide starting material.)

(4) (S) -4- (diethoxyphosphinyl) -3 - [[(1,1-dimethyl-)

ethyl) -diphenylsilyl3-oxy] butyric acid-methyl

ι e ster

A solution of 7.691 g of the iodide in 20 ml Triäthylphos- ¹ ^ phit is heated for 3.5 hours under argon in an oil bath at 155 ⁰ C. The mixture is cooled and excess phosphite is distilled off under reduced pressure of 0.5 mm Hg at 75 ° C. There remain about 8.0 g of yellow oil. The crude oil is purified by flash chromatography on 400 g of silica gel and eluted with hexane-acetone (4: 1). The product fractions are evaporated. Yield: 3.222 g (41.1 %) of the desired phosphonate title compound as a clear, colorless, viscous oil.

TLC (1: 1) hex-acetone, Rf = 0.51, U.V. and PMA.

In addition, 2.519 g (61.1 % corrected yield) of the starting iodide of part (3) is recovered.

(5) (s) -3 - [[(1,1-dimethylethyl) diphenylsilyl] oxy] -4-phosphonobutyric acid methyl ester

A solution of 9.85 g (20 0 mmol) of Phosphoant ^r .'eil B (4) in 20 ml CH ₃ Cl ₂ is washed successively with 5.31 ml (32.0 mmol, I, 5 eq.) Bistrimethylsilyltrifluoroacetamide (BSTFA) and 6.60 ml

(50.0 iflMol, 2.5 eq.) Trimetylsilyl bromide (TMSBr) treated 36

and the clear mixture at room temperature under argon approximately

i <: Ί / 8 8 6

- 48 -

2707 f4

Stirred for 15 hours. Then 80 ml of 5 % KHSO. added and the mixture extracted with ETOAc. The aqueous phase is back-extracted with NaCl saturated upo with EtOAc. The combined organic layers are washed with brine, over anhydrous Na ₂ SO. dried and evaporated under reduced pressure to the crude phosphonic acid title compound as a viscous oil

TLC / 7: 2: 1) 1PrOH-NH ₄ OH-H ₂ O, R f = 0.30, UV and PMA.

, (6) (S) -3 - [[(1,1-dimethylethyl) diphenylsilyl] oxy] -

4- (hydroxymethoxyphosphinyl) -butter3äure-ir.'ithyl ester

Approximately 20.0 mmol of crude phosphonic acid from part E (5) in 25 mL of dry pyridine is treated with 1/62 mL (40.0 mmol, 2.0 eq.) Dried ₃ O 3 screened CH ₃ OH and 4.54 g ( 22.0 mmol, 1.10 eq.) Of dicyclohexylcarbodiimide (DDC). The obtained

white suspension is stirred under argon at room temperature for about 15 hours. Then the pyridine is distilled off under reduced pressure and the mixture is then azeotroped with 2 times 15 ml of benzene. The residual oil is dissolved in EtOAc, filtered and treated with 1NO. HCl and brine, dried over anhydrous Na-SO _{4 2B} and evaporated under reduced pressure. Removing ^J 'yield: 8,272 g of crude ester title compound as an oil containing a small amount of precipitated dicyclohexyl urea (DCU) includes. ,

TLC (7: 2: 1) 1PrOH-NH ₄ OH H ₃ O, R f = 0.60, UV and PMA. j

F. (S) - '[t [4'-fluoro-3,3', 5-trimethyl- [1, diphenyl] -2-yl] -methoxyphosphinyl-S-tert-butyldiphenylsilyloxybutyric acid

me thylest ir

'5.595 g (about 14.7 mmol) of crude phosphonic acid monomethyl ester of Part ^E are dissolved in 30 ml of dry CH ₃ Cl ₂ , washed with

70 7 14

- 49 -

5.60 ml (29.4 mmol, 2.0 eq.) Of distilled trimethylsilyl. · Diethyl ester and stirred for 1 hour under argon at room temperature. Then the mixture is evaporated under reduced pressure, taken up with 30 ml of benzene and dried under reduced pressure. The pale yellow viscous oil is dissolved in 30 ml CH-Cl ₂ and 2 drops of 4 Α sieve dried DMF, the clear solution is cooled in salt / ice bath to -10 ⁰ C and dropwise through a syringe with 1.41 ml (16.2 mmol, 1.1 eq.) Of distilled oxalyl chloride.

It shows strong gas evolution and the solution gets a more yellow color. The mixture is stirred under argon for 15 minutes at -10 ⁰ C and then stirred at room temperature for 1 hour. Then the mixture is evaporated under reduced pressure, taken up with 30 ml of benzene, re-evaporated and dried under reduced pressure to give crude phosphonochloridate as a yellow oil.

A solution of about 14.7 mmol crude phosphonochloridate in 10 ml anhydrous CH ₂ Cl ₂ is added dropwise with a solution of 2.06 g (8.43 mmol) biphenyl alcohol of Part D in 15 ml anhydrous pyridine and the resulting mixture is stirred for 16 hours stirred under Arqon at room temerature q. The mixture is evaporated to dryness and the residue obtained is partitioned between 5% KHSO 4 solution and ethyl acetate. The organic phase is washed with saturated NaHCO ₃ solution and brine, over anhydrous Na ₃ SO 4. dried and evaporated under reduced pressure to give 8.290 g of brown oil. The crude product is purified by flash chromatography on 370 g of silica gel and elution with hexane-acetone (70:30). Product fractions are combined and evaporated. Yield: 3.681 g (66%) of the desired phosphonate title compound as a pale yellow oil

TLC (3: 2) hexane-acetone, R _f = v. ">, UV and PMA.

2707/4

- 50 -

G. (S) -4 - [[[4'-fluoro-3,3 ', 5-trimethyl-1- [1,1'-biphenyl] -2-yl- 1 - methoxyphosphiny1] -3-hydroxybutyric acid diethylester r

A mixture of 1.103 g (1.66 mmol) of silyl ether of Part F in 20.0 ml of anhydrous THF is added with 380 μΐ (6.64 mmol, 4.0 Aq of glacial acetic acid and 4.98 ml (4, 98 mmol,.), 0 Xq.) 1.0 M Tetrabutylammoniumfluoridlösuna treated and the clear crelbe Lösungunq under argon for about 15 hours at Re.umtemoeratür stirred. The mixture is partitioned between cold water and diethyl ether, the organic phase washed with saturated NaHCO ₃ solution and brine, dried over anhydrous Na ₃ SO ₄ and evaporated to give 1.174 g of a viscous yellow oil. The "crude oil is purified by flash chromatography on .47 g of silica gel and elution with CH ₂ C1 ₂ -acetone (85:15). The product fractions are evaporated. Yield: 679 mg (93.1%) of the desired alcohol title compound as a clear, viscous oil. TLC (1: 1) hexane-acetone, R _f = 0.41, UV and PMA.

H. (S) -4- [t [4-fluoro-3,3, 5-trimethyl- [1,1'-biphenyl] -2-yl] -methoxy] -methoxyphosphinyl] -3-hydroxybutyric acid monolithium salt

A solution of 184 ng (0.42 mmol) of methyl ester of Part G in 5.0 ml of dioxane is treated with 0.50 ml (1.2 eq of 1.0 N lithium hydroxide and the mixture stirred under argon for 3 hours at room temperature. The mixture is diluted with water, filtered through a 0.4 μΐη polycarbonate membrane and evaporated under reduced pressure, the residue obtained is dissolved in 75 ml of water, frozen and lyophilized The crude acid is dissolved in the minimum amount of water and placed on a 100 ml bed separated from resin CHP-20, eluting with a linear gradient of water / CH ₃ CN The product fractions are evaporated, the residue dissolved in 50 ml of water, filtered through a 0.4 μm polycarbonate membrane and lyophilized. Yield 174 mg (89 , 1%, by weight of the hydrate) of the desired monolithium salt title compound as a white solid.

51 TLC (7: 2: 1) IPrOH-NH ₄ OH-H ₂ O, R _f = 0.58, UV and PMA

Analysis for C ₂₁ H ₂₅ OgPFLi + 1, SS mol HO (MW 465.46).

CHFP

5: 54.19 6.26 4.08 6.65

F .: 54.19 6.21 4.29 6.43

H ¹ NMR (400 HMz):

61.74-2.08 ppm (2H, m, -Po (OCH ₃ ) CH ₂ '2.30 (3H, s, aromatic methyl group

2.32 (3H, d, aromatic methyl group in

to the

α-position fluorine itom, J .. "2, 2 Hz) 2,35-2,62 (2H, m, -CH ₉ CO ₇ Li) 2,46 (3H, s, aromatic methyl group) 3, '37 &

3.63 (3H, 2 doublets., -OP (OCH ₃ ) -, 2 diastereomers, J "= 10.3 Hz 4.28 (1H, m -CH ₂ CH (OH) CH ₂ CO ₂ Li)

4 20 4.97 (2H, nt PhCJI ₂ OP (OCH ₃ ) R)

6.87-7.25 (5H, m, aromatic H ₁ ¹ B)

Example 2

(S) -4- [[[4-fluoro-3,3 ', 5-trimethyl [(1,1-biphenyl) -2-yl] methoxy] -hydroxyphosphinyl] -3-hydroxybutyric acid-1,3-diol. msalz

A solution of 374 mg (0, O53 mmol) diester of Example "in 8.0 ml of dioxane is treated with 2.6 ml (3.0 eq. Of 1.0 N lithium hydroxide and under argon for 5 hours in oil bath at ^50?. C heated. in this case, a white precipitate shows. The mixture is diluted with water and filtered. The aqueous solution is extracted once with diethyl ether, filtered through a 0.4 micron polycarbonate membrane, and evaporated under reduced pressure. The crude product is purified by chromatography on resin CHP 20 (100 ml bed), using a linear gradient H ₂ O / CH ₃ CN

- 52-

is eluted. The product fractions are evaporated under reduced pressure, the residue taken up in 50 ml of water, filtered through a 0.4 μπι polycarbonate membrane and lyophilized. Yield 260 mg (67.1% by weight of the hydrate) of the desired dilithium salt title compound as a white solid

TLC (7: 2: 1). PrOH-NH ₄ OH-H ₂ O, R _f = 0.47, UV and PMA.

Analysis for C ₂₀ H ₂₂ OgPFLi ₂ +1.77 mol H ₃ O: 10 CHFP

Calc .: 52.88 5.67 4.18 6.82 Found: 52.88 5.26 4.24 6.43

H ¹ NMR (400 MHz, CD ₃ OD): 15 0

6 1.69 ppm (2H, m, -OPCH ₂ CH (OH) -)

2.26-2.42 (2H, m, CH ₂ CO ₂ Li)

2.30 (3H, s, ahydrous methyl group

2.31 (3H, d, aromatic methyl group in the ^ -position to a fluorine atom,

J _HF = 1.9 Hz)

2.38 (3H, s, aromatic methyl), gruDDe

4.22 (IH, m, -CH (OH) CH ₂ -)

0 ²⁵ 4.75 (2H, m, PhCH ₂ OP)

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

30 example

(3S) -4 - [[[4'-Fluoro-3,3 ·, 5-trimethyl [1,1'-biphenyl] -2-yl] -methoxy ] -methylphosphine-3-hydroxybutyric acid monolithium salt A. (S) -4 - [(Chloro) -methylphosphinyl] -3 - [[(1,1-dimethylethyl) -diphenylsilyl] oxy] -butyric acid methyl ester

The title of the phosphinchloride derivative becomes the first three

<- 53 -

paragraphs of Example 6 Part B produced.

B. (3S) -4 - [[(4'-PIuOr-S, 3 ', 5-trimethyl-Π / 1'-biphenyl] -2-yl] -methoxy] -methylphosphinyl] -3-tert-butyldiphenylsilyloxy Butyric acid diethyl ester

Anhydrous cooled in an ice bath to 0 ⁰ C solution of about 2.2 mmol Phosphinchloridat of part A and 429 mg (2.2 mmol) 1.0 Xq.) Biphenyl alcohol of Example 1, Part C in 10 ml ^CH 2 ^{C1 2 with 425 μ1} * ^{3 '04 1 01} ^' ^{1 '4 Å <3} · ^Et 3 ^{N and 27 m} 3 (0.22 mmol) of 4-DMAP. The orange solution is stirred under argon for about 15 hours at room temperature. Then the mixture is partitioned between 5% KHSO ₄ solution and ethyl acetate and the organic layer is washed with brine, dried over anhydrous Na ₃ SO ₄ and evaporated to give 1.1 g of an orange oil. The crude oil is purified by flash chromatography on 44 g of silica gel LPS-1 and elution with ethyl acetate: hexane (1: 1). The product fractions are combined and evaporated. Yield: 298 mg (21%) of the desired coupled title compound as a pale yellow oil. Also, 460 mg (67% corrected yield) of biphenyl alcohol starting compound of Example 1, part C (2) is recovered

TLC (1: 1) ethyl acetate-hexane, R _f = 0.18, UV and PMA.

C. Methyl (3S) -4 - [[[4'-fluoro-S, ¹ , 5-trimethyl- [1,1'-biphenyl] -2-yl ] -methoxy] -methylphosphinyl] -3-hydroxybutyric acid

A solution of 298 mg (0.46 mmol) of Part B silyl ether in 6.0 ml of anhydrous THF is added with 110 μΐ (1.84 mmol, 4.0 eq.) Of glacial acetic acid and 1.43 ml (3.1 eq. ) 1.0 M solution of tetrabutylammonium fluoride in THF. The resulting solution is stirred under argon for about 15 hours at room temperature. The mixture is then partitioned between cold water and ethyl acetate, and the organic phase is washed with saturated NaHCO3, brine, and anhydrous

ι

I -

XY.

2707 U

- 54 -

dried Na ₃ SO ₄ and evaporated to give 273 mg of yellow oil. The crude oil is purified by flash chromatography on 11 g of silica gel LPS-1 and elution with hexane-acetone (2: 2). The product fractions are combined and evaporated. Yield: 150 mg (80%) of the desired alcohol-titanium compound as a viscous oil

TLC (1: 1) hexane: acetone, R _f = 0.23, UV and PMA.

D. (3S) -4 - [[[4'-fluoro-3,3 ', 5-trimethyl- [1,1'-biphenyl] -2-yl] -methoxy] -methylphosphine] -3-hydroxybutyric acid mono

lithium salt

A solution of 150 mg (0.367 mmol) of methyl ester of part C in 3, 10 ml of dioxane is treated with 0.44 ml (1.2 eq of 1.0 N lithium hydroxide and the resulting white suspension is added under argon for 2 hours The mixture is diluted with water, filtered through a 0.4 μπι Poxycarbonatmembran and evaporated under reduced pressure to a colorless glassy material.

The crude product is taken up in the minimum amount of water and chromatographed on HP-20 (100 ml bed), eluting a linear gradient H ₂ O / CH ₃ CN. The product fractions are evaporated, the residue taken up in 50 ml of water, filtered through a 0.4 μπι polycarbonate membrane and lyophilislert. Yield: 130 mg (79% by weight of the hydrate) of the desired lithium salt title compound as a white solid

TLC (8: 1: 1) CH ₂ Cl ₂ -CH ₃ OH-HOAc, R _f = 0.52, UV and PMA.

Analysis for C ₃₁ H ₂₅ O ₅ FLiP + 1.73 mol H ₃ O (MW 445.49):

CHFP: 56.61 6.44 4.26 6.95

found: 56.67 6.36 4.31 7.43

't i, -. > j.

2 70 7 i

- 55 -

H ¹ NMR (400 MHz):

δ 1.49 ppm (3Η, d, -OP (CH ₃ ) -, J _Hp = 14.7 Hz)

O Il

1.83-2.0 (2H, m, P ₃ i ₂

2.27-2.40 (2H, m, CH ₂ CO ₂ Li)

2.30 (6H, s, 2 aromatic methyl groups)

2.44 (3H, s, aromatic methyl group

, 4.26 (IH, m, -CH ₂ CH (OH) CH ₂ CO ₂ Li)

4.87 (2H, m, ArCH ₂ OP (CH ₃ ) -)

6.90-7.20 (5H, in, aromatic protons)

. ·

". Example 4

(S) -4- [[i; 2,4-Dichloro-6- [(4-fluorophenyl) -raethoxy] -phenyl] -methoxy] -methoxyphosphinyl] -3-hydroxybutyric acid monolithium

salt

A « ? 4-dichloro-6- (4-fluorophenylmethoxy) benzaldehyde J. Med. Chem., 29 (1986), 167

A solution of 13.77 g (72.5 mmol) of 4,6-dichloro-2-h ^t> droxybenzaldehyd in 100 ml of DMF is added with stirring with 12.02 q (87 mmol) K ₂ CO ^. The mixture is heated 60 minutes to about 70 ⁰ C and then treated with 11.7 ml 4-Fluorbenzylbror.id. The resulting solution is stirred for 3 1/2 hours at 70 ⁰ C and then poured onto 1.5 liters of ice water, filtered, washed with water and recrystallized from diethyl ether / petroleum ether. Yield: 17.88 g (83%) off-white crystals, mp 107-108 ⁰ C.

B. 2,4-dichloro-6 - [(4- fluorophenyl) -methoxybenzenemethanol

10.0 ml in an ice bath to ⁰ ° C alpgekühlter anhydrous diethyl ether _t

1\

- 56 -

with 158 mg (. 4.16 mmol, 0.6 Xq) LiAlH ₄ are added and _d ie; gray suspension by dropwise addition of a solution of 2.06 g (6.39 mmol) of aldehyde from Part A in 10 mL of anhydrous THF treated. The mixture is warmed to room temperature and stirred under argon for 1 hour. Then the mixture is again cooled in an ice bath to 0 ⁰ C and the reaction is stopped by dropwise successive addition of 160 μΐ water, 160 μΐ 15% NaOH and 475 μΐ water. The precipitated salts are filtered through anhydrous Na ₂ SO ₄ over 1/4 "diatomaceous earth, the clear filtrate is evaporated to give 2.052 g (98.9%) of crude alcohol as white crystals, and trituration with cold hexane gives 1.892 g (m.p. 91.2%) pure alcohol title compound as a white crystalline solid of mp 72 to 73 ° C.

TLC f4: 1) hexane-acetone, R: = 0.31, UV and PMA. Analysis for .C ₁₄ H ₁ ^ ₂ Cl ₃ F (MW 301,142):

C H Cl F: 55.84 3.68 23.55 6.31 Found 55.97 3.71 23.42 6.30

C. (S) -4 - [[[2,4-Dichloro-6 - [(4-fluorophenyl) -methoxy] -phenyl] -methoxy] -methoxyphosphinyl] -3-tert-butyldiphsylsilyloxybutoxylate : : methyl ether

A solution of about 3.84 mmol of methyl ester of Example 1, part E (6). in 10 ml of anhydrous CH ₂ Cl ₂ is treated with 1.46 ml (7.68 mmol), 2.0 eq.) of distilled trimethylsilyldiäthylamin and the resulting solution was stirred at room temperature under argon for 1 hour. The mixture is evaporated under reduced pressure, azeotroped once with 20 ml of benzene and the residue is dried under reduced pressure. There is obtained crude silylated phosphonic acid monomethyl ester as a colorless oil.

A solution of about 3.84 mmol of the crude ester in 10 ml of anhydrous CH ₀ Cl ₉ and 1 drop of anhydrous DMF is added in ice /

"I t 1 " i

    ,. ,

r '

2707 f 4 '

- 57 -

Salt bath cooled to -1O ° C and treated dropwise with 368 μΐ (4.22 mmol, 1.1 eq.) Distilled oxalyl chloride. The clear yellow mixture shows gas evolution. The mixture is stirred at room temperature for 1 hour under argon, then evaporated under reduced pressure and azeotroped twice with 20 ml of 3-enol to give crude phosphonochloride as a viscous yellow oil.

Approximately 3.84 mmol of crude Phosphonochloridat anhydrous in 10 ml CH ₂ Cl ₂ at 0 ⁰ C in an ice bath with 1.15 g (3.84 mmol, 1.0 eq.) Of alcohol from Part B and then with 805 μΐ (5 , 76 mmol, 1.5 eq.) Of Et ₃ N and 47 my (0.384 mmol, 0.1 eq.) Of 4-DMAP and the brown mixture stirred for about 15 hours at room temperature under argon. The mixture is partitioned between 5% KHSO ₄ solution and ethyl acetate, the organic phase washed with brine, over anhydrous Na ₂ SO ₄ . dried and evaporated to give 3.197 g of dark brown oil. The crude product is purified by flash chromatography on silica gel (160 g) and Elutior. with hexane-ethyl acetate (7: 3). The product fractions are combined and evaporated. Yield: 594 mg (21.1%) of desired phosphonate title compound as a yellow oil. In addition, 688 mq (52.4% corrected yield) alcohol starting compound is recovered from Part B. TLC (1: 1) hexane-acetone, R _f = 0.29, UV and PMA

D. (S) -4 - [[[2,4-Dichloro-6-L (4-fluoro-phenyl) -methoxy-phenyl] -methoxy.] -Methoxy-phosphinyl-3-hydroxy-hydroxybutyric acid methyl ester .

A solution of 578 mg (0.788 mmol) of Silyl ester of Part C in 8 ml of anhydrous THF is added with 180 μΐ (3.2 mmol, 4.0 eq.) Of glacial acetic acid and then with 2.36 ml (2.36 mmol, 3, 0 eq.) 1.0 M solution of n-Bu ₄ NF in THF and the resulting pale yellow solution stirred for about 15 hours under argon at room temperature.

The mixture is then poured into cold water and extracted twice with ethyl acetate. The organic phase is treated with

2707 f 4

- 58 -

saturated NaHCO ₃ solution and brine, over

anhydrous Na ₂ SO. dried and evaporated to give 625 mg of yellow oil. The crude product is purified by flash chromatography on 31% silica gel and elution with hexane-acetone (7: 3). The product fractions are combined

and evaporated. Yield: 339 mq (86.9%) of the desired alcohol title compound as a clear, colorless, viscous oil

TLC (1: 1) hexane-acetone, R _f = 0.25, UV and PMA).

E. (S) -4 - [[[2,4-Dichloro-6 - [(4-fluoro-phenyl) -methoxy] -phenyl] -methoxy-methoxyphosphinyl] -3-hydroxybutyric acid mono- lithium salt -_

A solution of 132 mg (0.267 mg of Parti II phosphonate in 2.5 ml of dioxane is treated with 0.32 ml. 1.2 Xq.) 1.0 N lithium hydroxide and the mixture stirred under Arqon at room temperature for 4 hours , This creates a white precipitate. The mixture is diluted with water, filtered and the filtrate evaporated to dryness under reduced pressure. The residue is chromatographed on resin HP-20 (100 ml bed) and eluted with a linear gradient H ₂ O / CH ₃ CN. The product fractions are combined and evaporated, the residue taken up in water through a 0.4 μΐη. Pol.ycarbonatmembran filtered and lyophilized. Yield:. 108 mg (79%, by weight of the hydrate) of the desired lithium salt title compound as a white solid.

TLC (20: 1: 1) CH ₀ Cl ₀ - CH ₂ OH-HOAc, R- = 0.41, UV and PMA.

Δ Δ ο Γ

Analysis for C _Q H ₁ O ₇ Cl_FLi _o P + 1.42 mol H ₀ O (MW 511.72):

on \ J \ OI Δ Δ Δ

^dU CH Cl FP

Calc .: 44.59 4.10 13.86 3.71 6.05 Found: 44.22 4.09 13.91 3.72 6.11

- 59 -

H ¹ NMR (400 MHz):

O 6 1.98-2.11 ppm (2H, m, OP (OCH ₃₎ CH ₂ CH (OH) ppm -2.26-2.45 (2H, m, -CH (OH) CH ₂ CO ₂ Li) 3.63 &

3.62 (3H, 2 Qübletts,, 2 Diastero-

₀ mere)

OP * (OCH ₃ ) CH ₂ -, J ₁ Jp-II HZ)

4.23 (IH, m, (-CH ₂ CH (OH) CH ₂ CO ₂ Li) 5.16 (2H, s, F-PhCH ₂ O)

5.24 (2H, d, ArCH ₂ OP, JjJp = O.2 Hz). 7.13-7.53 (6H, m, aromatic H's)

Example 5 15

(3S) -4- [[[2,4-DiChior-O - [(4-fluorophenyl) -methoxy] -phenyl] -methoxy] -hydroxyphosphinyl] -S-hydroxybutyric acid dilithium salt

A mixture of 210 mg (0.424 mmol) of Part D diester in 4.0 ml of dioxane is treated with 1.30 ml (3.0 equiv.) Of 1.0 N lithium hydroxide and the colorless solution is treated in an oil bath under argon for 3.5 hours heated to 50 ⁰ C. After 15 minutes, a white precipitate appears. The mixture is diluted with water, filtered and the filtrate evaporated under reduced pressure. The residue is dissolved in the minimum amount of water and chromatographed on resin HP-20 (100 ml bed) and eluted with a linear gradient H ₂ OZCH ₃ CN. The product fractions are combined and evaporated. The residue is taken up in 50 ml of water, filtered through a 0.4 μπι polycarbonate membrane and lyophilized. Yield: 175 mg (81%) by weight of the hydrate) of the desired dilithium a.lz title compound as a white solid.

2707/4

TLC (8: 1: 1) CH ₂ Cl ₂ -CH ₃ OH-HOAc, R | = 0.07, UV and PMA.

Analysis for C H _{iC 1Q} O_Cl.FLi-P + 1.70 mol H ₀ O (MW 509.62):

C HF Cl P

: 42.42 3.84 3.73 13.91 6.08

F .: 42.46 3.90 3.93 13.42 5.66

H ¹ NMR (400 MHz):

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

2.27 (IH, dd, -CH (OH) OI ₂ CO ₂ Li, J ₁₀₁ = O.8

Hz)

2.39 (IH, dd, -CH (OH) OI ₂ CO ₂ Li, J _HH = 4.4

Hz)

4.26 (IH, m, CH ₂ CH (OH) CH ₂ CO ₂ Li)

5.08 (2H, s, F-Ph-CH ₂ OAr)

7.03-7.53 (6H, m, aromatic H's)

Example 6

(3S) -4 - [[2,4-dichloro-6 - [(4-fluorophenyl) methoxy] phenyl] -

methyl methoxy] -methylphosphiny 1] -3-hydroxybutyric acid A. (S) 3 - [[(1,1-dimethylethyl) diphenylsilyl] oxy] -4- ( ethoxymethylphosphine 1) -but-1-fluoroethyl ester

A mixture of 4.68 g (9.18 mmol) of iodide of Example 1, Part E (3) in 5.0 g (36.7 mmol) Methyldiäthoxyphosphin (Strem Chemicals) for 2.5 hours in oil bath at 100 ⁰ C and then heated to 150 ⁰ C under argon for a further 3 hours. The yellow solution slowly forms a white precipitate.

Excess phosphine is distilled off under reduced pressure (0.5 mm Hg) and the crude product is purified by flash chromatography on silica gel elution with He.xan-acetone (65:35). The product fractions are combined and evaporated. Yield: 1.590 g (38%) of the desired phosphine ester compound (mixture of diastereomers) as a clear, viscous oil.

2 70 7 f 4

- 61 -

! TLC (3: 2) hexane-acetone, R _f (2-diastereomers) = 0.19 and 0.22, UV and PMA

E. (3S) -4 - [[2,4-Dichloro-6 - [(4-fluorophenyl) -methoxy] -phenyl] -methoxy] -methylphosphinyl] -3-tert-butyldiphenylsilyl

butyric acid methyl ester

A solution of 605 mg (1.3 mmol) of phosphine ester of Part A in 6.0 ml of anhydrous CH ₂ Cl ₂ is mixed with 280 μΐ (1.05 mmol) 0.8 eq.). Bis (trimethylsilyl) trifluoroacetamide ( BSTFA) and 210 μΐ (1.57 mmol, 1.2 eq.) Of trimethylsilyl bromide (TMSBr) and the resulting solution stirred at room temperature under argon for about 15 hours. Then 15 ml of 5% KHSO ₄ solution are added and the mixture is extracted with ethyl acetate.

j5 The organic phase is washed with brine, over anhydrous Na ₂ SO. qetrocknet and evaporated under reduced pressure, whereby crude phosphinic acid is obtained as a colorless oil.

2Q A solution of about 1.3 mmol crude phosphinic acid in 6.0 mL anhydrous CH ₂ Cl ₂ is added at 270 μΐ. 1.44 mmol, 1.1 eq.) Distilled Trimethylsilyldiäthylamin and the clear mixture stirred at room temperature for 1 hour under argon. The mixture is evaporated under reduced pressure, azeotroped once with 15 ml of benzene and the residue is dried under reduced pressure.

A cooled in an ice bath to 0 ⁰ C solution of about 1.3 mmol crude silylated phosphinic acid in 6.0 ml of anhydrous CH ₃ Cl ₂ and

One drop of DMF is treated dropwise via syringe with 130 μL (1.44 mmol, 1.1 eq.) of distilled oxalyl chloride. It shows gas evolution. The mixture is stirred at room temperature under argon for 1 hour and then evaporated under reduced pressure, azeotroped twice with 15 ml of benzene and the residue is dried under reduced pressure to give crude phosphinochloridate as a yellow oil.

707 (4 '

- 62 -

A solution of about 1.3 mmol of phosphinochloridate cooled to ⁰ ° C. in an ice bath and 392 mg (1.3 mmol) of alcohol from Example 1, part E (6) in 6.0 ml of anhydrous CH ₃ Cl ₂ is mixed with 275 μΐ (1.97 mmol, 1.5 eq.) Of Et ₃ N and 16 mg (0.13 mmol, 0.1 Xq.) Of 4-DMAP and the resulting yellow mixture stirred under argon at room temperature for about 15 hours. The mixture is partitioned between 5% KHSO ₄ solution and ethyl acetate, the organic phase washed with brine, dried over anhydrous Na ₃ SO ₄ and evaporated to give 908 mg of crude product as a dark yellow oil. The crude product is purified by flash chromatography on 45 g of silica gel and elution with hexane-ethyl acetate (3: 2). The product fractions are combined and evaporated. Yield: 266 mg (28.3% desired title compound as a clear, colorless oil) Further, 197 mg (57% corrected yield) of starting alcohol compound are recovered.

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

A solution of 275 mg (0.38 mmol) of Part B silyl ester in 6.0 mL of anhydrous THF is added with 90 μΐ (1.53 mmol, 4.0 eq.) Glacial acetic acid and 1.2 mL (3.1 eq. ) 1.0 M solution of tetrabutylammonium fluoride in THF. The resulting solution is stirred under argon for about 15 hours at room temperature. Then the mixture is partitioned between cold water and ethyl acetate, the organic phase washed with saturated NaHCO ₃ solution and brine, dried over anhydrous Na ₃ SO ₄ and evaporated to give 258 mg of yellow oil. The crude product is purified by flash chromatography on 8 g of silica gel LPS-1 and elution with hexane-acetone (1: 1). The product fractions are combined and evaporated. Yield: 142 mg (77%) of the desired alcohol title compound as a clear, colorless oil.

35 TLC (1: 1) hexane-acetone, R _f = 0.20, UV and PMA.

2707

~ "Dj" *

D. (3S) -4 - [[2,4-Dichloro-6 - [(4-fluorophenyl) -methoxy] -phenyl] -methoxy] -methylphosphinyl] -3-hydroxybutyric acid monolithium salt

A solution of 142 mg (0.296 mmol) of methyl ester of Part C in 3.0 ml of dioxane is treated with 0.36 ml (1.2 eq) of 1.0 N lithium hydroxide and the white suspension obtained under argon for 2 hours at room temperature touched. The mixture is diluted with water, filtered through a 0.4 μπ \ polycarbonate membrane and the filtrate is evaporated under reduced pressure. The crude product is dissolved in the minimum amount of water and chromatographed on resin HP-20 (100 ml bed) and eluted with a linear gradient H ₂ O / CH ₃ CN. The product fractions are combined and evaporated. The residue is taken up in water, filtered through a polycarbonate membrane and lyophilized. Yield: 93 mg (63% by weight of the hydrate) of the desired lithium salt title compound as a white solid. TLC (8: 1: 1) CH ₂ Cl ₂ - CH ₃ OH-HOAc, R _f = 0.51, UV and PMA.

Analysis for C ₁₉ H ₁₉ O ₇ Cl ₂ FLiP + 1.38 mol H ₃ O (MW 495.94):

C H F Cl P ber. :: 46.01 4.42 3.83 14.30 6.24 Found: 46.10 4.49 3.82 14.32 6.43

H NMR (400 MHz)

1:53 ppm (3H, d, -OP (CH ₃₎ CH ₂ -, J _H _ _p = 14.6 Hz)

30, 0

1.87-2.10 (2H, m, -OP (OCH ₃ ) OI ₂ -)

2.27 (IH, dd, -CH (OH) CH ₂ CO ₂ Li,

J _H _ _H = 8.4 Hz, J _H _ _p = 1 Hz) 2.38 (IH, dd, -CH (OH) CH ₂ CO ₂ Li, ^J HH ^{= 4} * ^{7 Hz} ' ^J HP ^{Bl # 1 HZ} *

j Z70714

, - 64 -

4.29 (IH, m, -CH ₂₂₂

5.16 + 5.18 (4H, m, ArCH ₃ OP and F-PhCH ₂ O) -)

7.11-7.52 (6H, m, aromatic

Example 7

(S) -4 - [[[4'-Fluoro-3,3 ·, 5-trimethyl- [1,1'-biphenyl-2-yl] -methyl] -amino] -methoxyphosphinyl] -3-hydroxybutyric acid -

monolium salt :

A. 4'-fluoro-3,3 ', 5- trimethyl- [1,1'-biphenyl] -2-carboic acid

A solution of 1.0 g (4.13 mmol) aldehyde from Example 1,

Part C (2) in 10 ml of acetone is added dropwise in an ice bath at 0 ^° C.

with 4.1 ml (8.0 N, excess) of Jones reagent. The resulting green-brown suspension is stirred under argon for 15 hours at room temperature, excess oxidizing agent is decomposed by addition of 10.0 nil of isopropanol / and

• fallen chromium salts are removed by adding the suspension 20

The filtrate is evaporated, taken up in EtOAc, washed twice with saturated NH ₄ Cl and brine, washed twice with saturated NH ₄ Cl and brine, then dried over anhydrous Na ₂ SO ₄ and evaporated Yield:

1.011 g of a green solid, mp 153-154 ° C. The raw 25

Acid is purified by dicyclohexylamine acid. To a

(Solution of 5.0 mL crude acid in EtOAc add 823 μΐ (1.0 eq) dicyclohexylamine (DCHA) Dilute the solution with hexane and collect the precipitated crystalline salt Yield: 997 mg (55% from the aldehyde , Mp 181-183 ° C) of the desired product as eggshell-colored, crystalline DCHA salt.

The free acid of the title compound is recovered from the DCHA salt by dissolving the salt between 5% KHSO ⁴ and EtOAc ⁴

is split. The organic phase is washed with brine, dried over anhydrous Na ₃ SO ₄ and placed under

2707/4

.1 - 65 -

evaporated under reduced pressure. Yield: 554 mg (52% from the aldehyde) of the desired acid title compound. TLC (9: 1) CH ₂ Cl ₂ -CH ₃ OH, R _f = 0.37, UV and PMA.

4'-fluoro-3,3 ', 5-trimethyl-fi, 1'-biphenyl] -2-carboxamide

A solution of 554 mg (2.14 mmol) acid from Part A in 6.0 mL dry CH ₃ Cl ₂ and dry (1 drop) DMF is added dropwise in an ice bath (0 ⁰ C) via a syringe containing 205 μl (2 , 35 mmol, 1.1 eq.) Of distilled oxalyl chloride. The clear yellow solution is stirred under argon for 1 hour at room temperature. The mixture is evaporated under reduced pressure, azeotroped twice with benzene and dried under reduced pressure to give crude acid chloride as a yellow oil.

A cooled in an ice bath (0 ⁰ C) mixture of 3.0 ml of THF and 2.0 ml (excess) of concentrated NH.OH is added dropwise with 3.0 ml of a THF solution of the tubular acid chloride.

^{£ KJ} The light orange solution is stirred under argon for 1 hour at room temperature. The mixture is partitioned between water and EtOAc, the organic phase washed with saturated NaHCO 3, brine, water and brine, then dried over anhydrous Na ₃ SO ₄ and evaporated. Yield: 528 mg

^ ° (96.1%) crude amide as helloranqer solid. After one-time recrystallization from EtOAc-hexane, 435 mg (79.1%) of the purified amide title compound are obtained as pale yellow needles of mp 197-19 UV and PMA.

F. 197-198 ° C. TLC (1: 1) Et ₂ O-acetone, R _f = 0.83,

C. 4'-fluoro-3,3 ', 5-trimethyl- [1,1'-biphenyl] -2-methanamine

5.0 mL dry in a water bath (0 C ⁰⁾ is chilled THF with 125 mg (3.3 mmol) solid LiAlH ₄ was added. The gray suspension is added dropwise over 5 minutes with a solution of 424 mg (1.65 mmol) of Part B amide in 5.0 ml of THF. The

2 70 7 f 4

- 66 -

The suspension obtained is stirred under argon for 2 1/2 hours at room temperature and then heated for 45 minutes under reflux. The mixture is cooled in an ice bath to C ⁰ C and terminated by the dropwise successive addition of 125 μΐ H ₂ O, 125 μΐ 15prozentiger NaOH solution and 375 μΐ H ₂ O terminated. Precipitated aluminum salts are removed by filtration through anhydrous Na ₃ SO ₄ on kieselguhr. The clear filtrate is evaporated under reduced pressure and crude amine is obtained as a clear oil.

TLC (7: 3) Et ^ -acetone, R _f = 0.60, UV and PMA. The amine is purified as the HCl salt.

To a solution (about 1.65 mmol) of the crude amine in 8.0 ml of absolute EtOH is added 152 μ l (1.82 mmol) of concentrated HCl. The mixture is stirred under argon for 15 minutes at room temperature. The mixture is evaporated under reduced pressure to a white, crystalline solid. The solid is triturated with cold Et-O, collected by filtration and dried under reduced pressure. Yield:

426 mg (92.4%) of the amih HCl title compound as fine white crystals.

D. (S) -4 - [[[[4'-Fluoro-3,3 ·, 5-trimethyl- [1,1 · biphenyl] -2-yl] -methyl] -amino] -methoxyphosphinyl] -S tert-butyldiphenyl silyloxybutyric acid methyl ester

A solution of about 2.0 mmol of methyl ester of Example 1, part E (6) in 5.0 ml of dry CH ₃ Cl ₂ is treated with 758 μΐ (4.0 mmol, 2.0 Aq.) Of distilled trimethylsilyl diethylamine. The clear mixture is stirred under argon for 1 hour at room temperature. The mixture is evaporated under reduced pressure, azeotroped once with 15 ml of benzene and dried under reduced pressure.

A cooled (0 ⁰ C) solution of crude silyl phosphonate in 7.0 ml of dry CH ₃ Cl ₂ and DMF (1 drop) is added dropwise comparable distilled at 192 μΐ (2.2 mmol, 1.1 eq.) Of oxalyl chloride

2707 (4 '

- 67 -

puts. It shows gas evolution from the clear yellow mixture. The solution is stirred for 1 hour at room temperature, evaporated under reduced pressure, azeotroped twice with 15 ml of benzene and dried under reduced pressure to give crude phosphonochloridate as a yellow viscous oil.

A cooled (0 ⁰ C) solution of · Phosphonchloridat and 416 mg (1.49 mmol) of Part C BLphenylamin HCl in 10 ml of dry ^CH 2 ^{C1 2 with 641 μ1} * ^{4 '6 mMo1' 2 '3 A (3} x ) ^Et 3 ^{N and 24 m <} 3 (0 lbs. 2 mmol, 0.1 Xq.) Of 4-DMAP. The clear yellow mixture is stirred under argon for 15 hours at room temperature. The mixture is partitioned between 5% KHSO _d and EtOAc, the organic phase washed with brine and then over anhydrous Na 2 SO 4. dried and evaporated under reduced pressure. Yield: 1.19 g of yellow oil. The crude product is purified by flash chromatography on 60 g of silica gel and eluted with hexane-acetone (7: 3). The product fractions are evaporated. Yield: 588 mg (59.5%) of the phosphonamide title

20 compound as pale yellow, viscous oil.

TLC (7: 3) hexane-acetone, R _f = 0.20 UV and PMA.

E. (S) -4 - [[[[4'-fluoro-3,3 ', 5-trimethyl- [1,1'-biphenyl] -2-yl] methyl] amino] methoxyphosphinyl] -3 hydroxybutyrate methyl ester

A solution of 588 mg (0.888 mmol) of Silyl Ether of Part D in dry 10.0 ml of THF is added with 203 μΐ (3.55 mmol, 4.0 Xq.) Of glacial acetic acid and 2.66 ml (2.66 mmol, 3 , 0 Xq.) Of a 1.0 M tetrabutvlammonium fluoride containing THF solution. The resulting solution is stirred under argon for 15 hours at room temperature. The mixture is poured into cold H ₃ O and extracted with EtOAc. The organic phase is washed with saturated NaHCO ₃ solution and brine, then dried over anhydrous Na ₃ SO ₄ and evaporated under reduced pressure. Yield: 605 mg of orange oil. The crude product is purified by flash chromatography

2 70 7 ί 4 '

- 68 -

purified on 36 g of silica gel and eluted with hexane-acetone (1: 1). Product fractions are combined and evaporated. Yield: 196 mq (50.4%) of the title alcohol compound as light orange O]. TLC (1: 1) hexane-acetone, R _f = 0.16, UV and

5 PMA.

F. (S) -4 - [[[[4-Fluoro-3,3 ', 5-trimethyl- [1,1 · biphenyl] -2-yl] -methyl] -amino] -methoxyphosphinyl] -3- hydroxybutyric acid monolithium salt

To a solution of 105 mg (0.240 mmol) of diester of Part E in 2.0 ml of dioxane is added 288 μl (1.2 eq.) Of 1.0 N LiOH. The white suspension is stirred for 4 hours under argon at room temperature , The mixture is diluted with H ₂ O, filtered, and the filtrate is evaporated under reduced pressure. The residue is chromatographed on resin HP-20 (100 ml bed) and eluted with a linear gradient H ₂ O / CH ₃ CN. The product fractions are combined and evaporated. The residue ... is taken up in 50 ml of H 2 O, filtered through a 0.4 μm polycarbonate membrane and lyophilized.

Yield: 70 mg (62.7% on a hydrate weight basis) of the lithium salt title compound as a white solid. TLC (20: 1: 1) CH ₂ Cl ₂ -CH ₃ OH-HOAc, R _f = 0.19, UV and PMA.

^ Analysis for C ₂₁ H ₂₆ NO ₅ PFLi + 2.41 mol H ₃ O (MW 472.75):

53 C 6 H 2 N 4 F 6 P calc .: 53 35 6 , 57 2 96 4 , 02 6 , 55 gef. : 35 , 52 , 98 , 05 , 59

30 H ¹ NMR (400 MHz):

0 δ 1.79-1.97 ppm (2H, m, - ₃₂

2.26-2.44 ppm (2H, ra, -CH ₃ CO ₂ Li 2.29 (3H, s, aromatic methyl group) 2.31 (3H, d, aromatic methyl

group in α-position to the fluorine atom J ₁₄ F ^ I / ⁴ Hz) 2,47 (3H, aromatic Mechy! group)

2 70 7

- 69 -

3.46 and 3.50

3.96

4.17

6.84-7.21 (3H, 2 doublets: .2 diastereo-

mers, J _Hp = 10.5 Hz)

O Il

(2H, m, -PhCH ₂ NHPlOCH ₃ ) - (IH, m, (-CH ₂ CH (OH) CH ₂ CO ₂ Li)

(5H, m, aromatic protons)

Examples 8 to 20

According to the methods described above, the following additional compounds can also be prepared.

0 HI 'I RP -CH ₂ -C -CH ₂ -CO ₂ -R ^x

X OH ι

Ex

 Mr. R.

-

8. OH

9. C ₂ H ₅ O

10. C ₃ H ₇

CH.

Cl

^cl

C ₂ K ₅

η X R

1 0 H

2 NH CH.

10 Li

- 70 -

2707 f 4

Ex. η χ

11th

12. OH

2 NH H

2 0 H

13th

^ HJ

⁴ Ο) -CH ₂ -O

10 Li

C ₂ H ₅

14th

NH CH.

C ₂ H

₂ H ₅

15. OK

0 OK

c "o

CH.

- 71 -

Ex

 No.

16. ONa

to2 0

CH ₃ O

• ι

1 NH H

HO

18. CH.

C-H-C C O

CH ₃ CH ₃

OH1 0. H

19. HO

2 0 Ll

20. CH ₃ O

1 NH H

- 72 1 Example 21

(S) -4-Diisopropyloxyphosphinyl) -3 - [[(1,1-dimethylethyl) di

phenylsilyl] oxy] -butyric acid methyl ester

21.70 g (45.1 mmol) of iodide from Example 1 part E (3) are stirred for 30 minutes under greatly reduced pressure. 93.92 g (0.451 mol, 113.37 ml) of freshly distilled triisopropyl phosphite are added all at once and the reaction mixture is heated for 16 1/2 hours under argon at 155 ° C in an oil bath.

The mixture is then cooled to room temperature, excess triisopropyl phosphite and volatile reaction products are by short path distillation (10 mm Hg) and then the course of 8 hours at 100 ⁰ C by Kugelrohr distillation (0.50 mm Hg). Purify the product more finely by flash chromatography (9 mm diameter column) on silica gel (6 "Merck) and elute with hexane / acetone / toluene (6: 3: 1) in 50 ml fractions (2" / minute flow rate) , 17.68 g (33.96 mmol, 75% yield) of the isopropylphosphonate

20 title compound obtained as a clear viscous oil.

TLC: silica gel R _f = 0.32 (6: 3: 1 hexane-acetone-toluene).

¹ HNMR: (270 MH ₂ , CDCl ₃ ) 25 δ 7.70-7.65 (m, 4H)

7.45-7.35 (m, 6H)

4.57-4.44 (m, 3H)

3.59 (s, 3H)

2.94 and 2.88 (2xd, IH J = 3.7 Hz) 30 2.65 and 2.60 (2xd, IH 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)

2707 f 4

- 73 1 Example 22

(S) -4- (Hydroxymethoxyphosphiny1) -3- [[(1,1-dimethylethyl) diphenylsilyl] oxy] -butyric acid methyl ester dicyclohexylamine ^. (1; H salt

10.66 g (30.5 mmol) of isopropyl phosphonate from Example 21 are stirred in 80 ml of dry CH ₂ Cl ₂ under argon at room temperature. The solution is added dropwise over 5 minutes with 8.44 g (32.8 mmol, 8.71 ml) of bistrimethylsilyl trifluoroacetamide (BSTFA) followed by dropwise addition over 10 minutes of 7.84 g (51.3 mmol, 6.75 ml ) Trimethylsilyl bromide (TMSBr). After stirring for 20 hours at room temperature, the reaction is terminated by the addition of 200 ml of 5% aqueous KHSO. stopped and the reaction mixture is stirred vigorously for a further 15 minutes. The aqueous phase is extracted three times with ethyl acetate. The organic extracts are combined, washed once with brine, dried over Na ₃ SO ₄ and concentrated under reduced pressure. The resulting residue is azeotroped twice with 50 ml of toluene. The precipitate formed is suspended in toluene and then filtered. The filtrate is concentrated and distilled azeotropically again and filtered. The resulting filtrate is first evaporated under reduced pressure and then for 5 hours under high reduced pressure. The resulting viscous clear oil is stirred in 50 ml of dry pyridine under argon at room temperature. 4.65 g (22.6 mmol) of dicyclohexylcarbodiimide (DCC) and then 1.31 g (4 ', 0 mmol, 1.67 ml) of methanol are added all at once to this solution. After stirring for 20 hours at room temperature, the reaction mixture is filtered through diatomaceous earth in a glass funnel. The diatomaceous earth is washed with ethyl acetate and the combined filtrates are eingriamoft under reduced pressure. The residue is redissolved in ethyl acetate and washed twice with 5% aqueous KHSO ₄ and once with brine. The organic extract is dried over Na ₅ SO ₄

2707 f4

- 74 -

and filtered. The filtrate is concentrated and azeotroped twice with toluene, then taken up in toluene and filtered. The resulting filtrate is concentrated again, distilled azeotropically, filtered and evaporated first under reduced pressure and then for 6 hours under high reduced pressure. Yield: 10.2 g (more than 100% yield) Phosphonate monoester as a clear, viscous oil. TLC: silica gel R _f = 0.50 (7: 2: 1 nPrOH / NH ₄ OH / H ₂ O). 1.21 g of phosphonate monoester are evaporated under strongly reduced pressure for 4 hours to give 1.16 g (2.57 mmol), which are then dissolved in 10 ml of dry ethyl ether, followed by dropwise addition of u, 481 g (2.65 mmol , 0.528 ml) of dicyclohexylamine are added. The resulting homogeneous solution is allowed to stand for 7 hours at room temperature, whereby significant crystal formation occurs. The chamber is kept for 16 hours at -20 ⁰ C and then warmed to room temperature and filtered. The crystals are washed with cold, dry ethyl ether and then 18 hours under greatly reduced pressure over P ₂ ⁰ S ^e i ^{n e} 9 ^e d ^at Pft. The crystals are then evaporated under a reduced pressure for 4 hours at 45 ° C to give 1.25 g (1.98 mmol, 77% yield) of the dicyclohexylamine salt title compound as a white powdery solid, mp 155-156 ° C , TLC: silica gel R _f = 0.57 (20% MeOH (CH ₂ Cl ₂ ) ¹ H NMR:

25 (270 MH ₂ , CDCl ₃ ).

2707 f4

- 75 -

7.71-7.65 (m, 4H)

7.40-7.32 (m, 6H)

4.0.2 (m, IH)

3.52 (s, 3H)

3.28 umd 3.22 (m, IH)

3.11 (d, 3H J = Il Hz)

2.77-2.64 (m, 2H)

2.62-2.56 (m, IH)

1.92-1.08 (series fromm, 22H) 10

1.00 (S, 9H)

Mass Spectrum (FAB) 632 (M & H) IR: (KBr) 3466-3457 (wide)

3046, 3016, 2997, 2937, 2858, 2836, 2798, 2721, 2704, 2633, 2533, 2447, 1736, 1449, 1435, 1426, ¹⁵ 1379, 1243, 1231, 1191, 1107, 1074, 1061, 1051,

CM-I

analysis for C ₂₂ I * 31 ° 'GPSI ^<C 1 2 ^H 23 ^N I: 2 N, 22 20 Ber. : 64 C, 63 8th H, 6 1 2 , 18 Gef .: 64 , 51 8th , 49

25

30

35

Claims (5)

V "- 76 - ; | 1 claims j 1. A process for the preparation of compounds of the formula i. ' ^: i _e o ** 5 Jl RP-CH ₂ -CH-CH ₂ -CO ₂ R ^X X 6h (D <? ^H 2 ^J n
2 10 in the R, a hydroxyl group, a lower alkoxy or * Is lower alkyl; X is an oxygen atom or the group -NH-; η is 1 or 2 , Z is a hydrophobic anchor; , R ^{x is} a hydrogen atom or a lower alkyl radical, in the form of the free acid or in the form of a physiologically hydrolyzable and compatible ester or salt, characterized in that a silyl ether of the formula R ^C -? CHO CH-CHo-CO. alkvl ' XO
(CH ₂ J _n Si-C (CHa) ₃ ZC ₄ H ₅ V ₅ H ₃ '· C in which R is a lower alkyl or lower alkoxy radical, Silylätherspaltung subjects to an ester of the formula 30 0 " r . R ^- P-CK ₂ -CH-CHo-CO ₂ a x Be 35 ,1 27E7 14 - 77 - in which R * is an alkyl radical, and optionally the ester in the corresponding salt of the formula RP-CH ₂ -CH-CH ₂ -CO ₂ alkali metal OH (CHO) ι " ¹⁰ in which R is a lower alkyl or lower alkoxy radical, or in the acid of the formula RP-CH ₂ -CH-CH ₂ -CO ₂ K. X OH 15 ' (CH ₂ J _n or in the corresponding salt of formula 20 Ö Il AlkalimetallO-P-CHO-CH-CHO COoAlkalimetall l - = X OK 25
(or in the acid of the formula Il HO-P-CH ₂ -CH-CH ₂ -COO H X OK 30 ι (CH ₂ ) transforms.
35 70 714 2; 70 7 f 4 - 12a - 2. The method according to claim 1, characterized in that the hydrophobic anchor represents a lipophilic residue which, when pi · is bound to the HMG-like upper side chain of the molecule via a suitable bridging ligand, connects to a hydrophobic site of the enzyme, which is not used to bind the substrate HMG CoA. 3. The method according to claim 1, characterized in that Z is one of the radicals of the formulas, OAER alkvl (the dotted lines are possible double bonds) in which R, R, R ^a and R are the same or different and are independently hydrogen, halogen, lower ralkyl-, haloalkyl, phenyl odf · - substituted phen ¹ ireste or denote radicals of the formula 0R ^y, where R ^{y is} a Represents hydrogen, alkanoyl, benzoyl, phenyl, halophenyl, phenyl-lower alkyl, lower alkyl, cinnamyl, haloalkyl, allyl, cycloalkyl-lower alkyl, adamantyl-lower alkyl or substituted phenyl-lower alkyl; r ' 10 15 20 270 7 j 4 .. - 79 if Z is the remainder of the formula alkyl 5 5 '
the radicals R and R are identical or different and represent hydrogen atoms, lower alkyl radicals or hydroxyl groups. mean,
0 R ^{6 is} alkylC or arylCH, - is, R represents a lower alkyl radical, a hydroxyl or oxo group or a halogen atom, q is 0, 1, 2 or 3, in Fonr. the free acid or in the form of a physiologically hydrolyzable and compatible ester or salt. 4. The method according to claim 3, characterized in that X is an oxygen atom, R is an alkoxy radical and Z has the structure 2530 35 5. The method according to claim 3, characterized in that X is the group -NH-, R is an alkoxy radical and (CH ₀ ) CH ₀ and ΔTi Δ Z has the structure \. _r , '/. Γ, h ο - L. CJG; δ. Process according to claims 1 to 5, characterized in that the following compounds of general formula I are prepared: (S) -4-i [[4'-fluoro-3 _/ 3 ^l , 5-trimethyl- [1 _/ 1'-biphenyl] -2-yl] -, ^v- methoxy] -methoxyphosphinyl] -3-hydroxybutyric acid methyl ester or their monolithium salt, (S) -4- [[[4-fluoro-3,3 ', 5-trimethyl- [1,1'-bipheny] 1-2-yl] -methoxy] -hydroxyphosphinyl] -3-hydroxybutyric acid dilithium salt ,: . (3S) -4 - [[[4'-fluoro-3,3 ·, 5-trimethyl- [1,1'-biphenyl] -2-yl] methoxy] -methylphosphinyl] -3-hydroxybutyric acid monolithium , salt, (S) -4 - [[[2,4-dichloro-6 - [(4-fluorophenyl) ^ methoxy] phenyl] methoxy] -methoxyphosphinyl] -3-hydroxybutyric acid-mono lithium salt, (3S) -4 - [[[2,4-dichloro-6 - [(4-fluorophenyl) methoxy] -phenyl] -, 25 methoxy] -hydroryphosphinyl] -3-hydroxybutyric acid dilithium salt, (3S) -4 - ([2,4-dichloro-6 - [(4-fluorophenyl) -methoxy] -phenyl] -methoxy] -methylphosphinyl] -3-hydroxybutyric acid or its methyl ester or (S) -4-t {[(4'-fluoro-3,3 ', 5-trimethyl (1,1'-biphenyl-2-yl] methyl] amino] -methoxyphosphinyl] -3-hydroxybutyric acid monolithium salt.
35 5 I! RP-CH ₂ -CH-CH ₂ -CO ₂ R ^X X OH «R ^H 2> n Z · ' wherein R is a hydroxyl group, a lower alkoxy or lower alkyl group; " X is an oxygen atom or the group -NH-; η is 1 or 2,
Z is a hydrophobic anchor; R ^{x is} a hydrogen atom or a lower alkyl radical, in the form of the free acid or in the form of a physiologically hydrolyzable and compatible ester or salt, which is characterized in that a silyl ether of the formula