AU2778500A - Tetrahydrofuran antifungals - Google Patents

Description

C.

C C

C.

C *9

C.

CC**

C

0*

C.

C

CeaC

C

AUSTRALITA

Patents Act 1990 COMPLETE

SPECIFICATION

STAN~DARD

PATENT

Applicant SCHERING

CORPORATION

Invention Title: TETRA1YDROFUPAN ANT IFUNGALS The following statement is a full description of this invention, including the best method of performing it known to me/us: TETAHDOU N ANTIFUNGALS BACKGROUND OFTE INE

DTON

be. This invention relates to tetrahydrofuran antifungals, (2R-cis)-4-[4- be be.

4 -dihalophenyl)tetrahydro5(l 1-1 2 ,4-triazol. 1 -ylmethyl)terayrourn3-llehoyphnyl2,-dhdr-2[str and ethers of mono- 9. or dihydroxy-substituted (04-05) alkyl]-3H-1 2 4 -triazol-3..one substituted antifungals, and salts thereof, pharmaceutical compositions containing them, and methods of treating and/or preventing antifungal infections in hosts, including warm-blooded animals, especially humans with such tetrahydrofuran oboe antifungals.

International Publication Number WO 89/04829, published 1 June 1990 and LISP 5,039,676 Saksena gi gi.) discloses a and ()(trans antifungai compounds represented by the formula xCH 2 O& \N/N-Z 0 x CH 2

N

wherein X= F, CI; Z=Ioweralkyl, (02-08) alkanoyl or phenyl substituted by 2 -loweralkyl-3-oxo-1 2 4 -triazoI-4-yI,e.g. and (±)-rans-l- 4 -[2-(2,4-dif uorophenyl-2[(l H1 2 4 -triazol-1 l-yl)methylltetrahydro.4f uranyllm eth oxy]ph enyl- 4 -m ethyl eth yl)pipe razine. However, WO 89/04829 does not disclose the compounds of this invention.

COmmonly-owned European Patent Publication No. 05399381, published 5 May 1993 discloses, for example, [5)jia4[-4[-[-24 H-i 2 ,4-triazol- 1-ylmethyl) .tetrahydrofuran-3 7yljm ethoxylpheny]. 1 -pipe razinyljpheynyl)- 2 ,4-dihydro-2-(C 1 -Ci 1o)alIkyl)]-3H l, 2 4 -triazol-3-one antifungals but does not disclose the compounds of this invention.

Janssen U. S. Patent 4,791,111 discloses, for example, 4 4 -[U2-2,4..dichloroph en yl)2-(1 H 1, 2 triazol -1 -yl me th yl) 3-d ioxo la n-4 **yllmethoxylphenyl.. 1 -pipe razin yl]-2, 4 dihyd ro-2- (2hydroxy- I -methylpropyl)-3H.

1,24tizo--n useful as an antimicrobial aetand having increased solubility, but does not disclose the compounds of this invention.

Intemnational Publication Number WO 95/1 7407, published 29 0 0 0 June 1995, discloses tetrahydrofuran antifungals, 5 2 4 *dihalophenyltetrahydro5( 11:- 1 ,2,4-triazol- 1-ylm ethyl)..tetrahydrofuran.3- 0: yllm ethoxy]phenyl]..24.dihydro-2-[mono- or dihydroxy-substituted (03-08) 20 alkyl]-3H-.1,2, 4 -triazol-3..one substituted antaifungals, pharmaceutically acceptable esters, ethers and salts thereof, pharmaceutical compositions containing them, and methods of treating and/or preventing antifungal infections in hosts, including warm-blooded animals, especially humans with such tetrahydrofuran antifungals.

International Publication Number WO 95/19983, published 27 July 1995, discloses azole-methyl substituted dioxolanes and tetrahydrofurans and which are disclosed to. be water soluble and to have antifungal activity.

There is a need for broad-spectrum antifungal agents having increased solubility and having favorable activity profile for treating systemic fungal infections, especially AsD~illus, andida, Cvrptococcus and opportunistic infections.

SUMMARY OF INVENTION The present invention provides compounds represented by formula I H R /N o

N

*N

0 N "N-N

[I]

10 wherein X is independently both F or both Cl or one X is independently F and the other is independently Cl;

R

1 is a straight or branched chain (C4-5) alkyl group substituted by one or two groups convertible in vivo into hydroxy moieties or a pharmaceutically acceptable salt thereof.

Preferably the group R1 of formula I convertible in vivo into a hydroxy moiety, OH, is a polyether ester, phosphate ester or an amino acid ester.

In a preferred aspect of the present invention, there is provided compounds represented by formula

II

o R xN

[II

N

wherein X is independently both F or both Cl or one X is independently F and the other is independently Cl; wherein

R

2 is a straight or branched chain (C4-C5) alkyl group substituted by an amino acid ester groups convertible in vivo into a hydroxy moiety or a pharmaceutically acceptable salt thereof.

Preferably R2 in formula 11 is a straight or branched chain C5 alkyl group substituted by an amino acid ester group convertible in vivo into a hydroxy moiety.

In another preferred aspect, the present invention provides a compound represented by formula 111 .0 .F NN N 11

N

wherein

R

5 is 4S-Me 4 f 1 1 I

I

me/ RMe Me R S Me me S %%-Me or R 1 Mee

R,

M e

S/IM

Me R R.%%Me p- 7 Me Me ~Me RMe 1 4R 1 j Me SMe

R

1 SOOP Me

\-RI,

wherein

R

1 is an amino acid ester thereof or a pharmaceutically acceptable salt thereof.

In another aspect of the present invention there is provided a compound represented by the formula

IV

F NR9 0 =0 I wherein Rg wherin 9 =CH(C2HS)CH(R 6 )CH3 or CH(CH3)CH(R 6

)CH

3 wherein

R

6 is an amino acid ester group convertible in vivo into OH and the carbons with the asterisk has the R or S absolute configuration or a pharmaceutically acceptable salt thereof.

The preferred group R6 of formula IV convertible in vivo into OH is an amino acid ester group.

D ILD N F THE INVNON N

PREF

EMBODIMENTS

The term "(C4-C5) alkyl group substituted by one or two hydroxy moieties", as used herein means straight and branched chain alkyl groups of four or five carbons including but not limited ton- and i.g- i- and itbutyl, and S a~ and l.-pentyl substituted by one or two hydroxy moieties and includes R and S stereoisomers of such (C4-C5) alkyl groups.

The term "hydroxy-substituted C4 or C5 alkyl group" as used herein means -CH(C2Hs5)CH(OH)Ci3,-CH(C 2

H

5 )CH2CH2OH,

-(CH)-CH(OH)C.H,

20 CH(CH)CH(OH)CH,

-CH(CH

3 )CH(OH)C or -CH(C.Hs)CH20

H

wherein each carbon with the asterisk has the R or S absolute configuration.

The term "group convertible in vivo into OH" means a group transformable in iv into a hydroxyl group by e.g. hydrolysis and/or by an enzyme, e.g. an esterase. Such groups include polyether esters, phosphate esters, sulfate esters, heterocyclic esters, alkanoate esters, alkenoate esters, amino acid esters and acid esters. Preferred groups convertible in v into a hydroxyl group are the polyether esters, phosphate esters and amino acid esters.

The term "ethers" means straight and branched chain alkyloxy groups of one to twenty carbons, preferably of one to eight carbons, more preferably one to six carbons; polyethylene glycols, e.g. PEG200 to PEG 10000, preferably PEG200 to 5000; and (C1-C6) alkoxy or aryl(C 1

-C

6 alkoxy groups of the formula -O-(CHR17)g-Ar wherein

R

17 is (C1-C6) straight and branched chain alkyl and g= 0 to 6, preferably 1 to 3 and Ar is phenyl, phenyl substituted by halo, especially chloro and fluoro, or by nitro, cyano and trihalomethyl especially trifluoromethyl. The (C1-C6)( alkoxy or aryl (C1-C6)alkoxy groups which are conveniently made by the well known Williamson ether Synthesis. Typically suitable ether groups include methoxy and benzoxy.

xy The term "esters" means polyether esters phosphate esters S(c) heterocyclic esters alkanoate and alkenoate esters amino-acid esters acid esters and sulfate esters.

The term "polyether esters" as used herein means those polyether esters represented by the formula C (CHR)s-(O CHR )-OR wherein R* D w herein

R

7 is a straight or brancahed chain (C1-C6) alkyl group or hydrogen and s is an integer from 1 to 6, preferably s 1 to 3 and more preferably s 1; t is an integer from 1 to 6; preferably t is 1 to 3, more preferably t is 2 or 3.

R is R7 or (CHR)s- CO2R7 -CHs R 7 or preferably R 8 is CH 3 or C2H 5 or CHCOH r -CH2C0 2

CH

3 orTypically suitable polyether esters include -COCH20(CH 20), CH 3

-COCH

2 0(CH 2

CH

2 2

CH

3 2

CH

2 0)3CH ,and The term 'phosphate esters" as used herein means those phosphate acids esters represented by the formula -0C HR7)

(OW)

2 or 0 -OC 1(CHR7)n

(CHR

7 )f 1 11 P(OW) ,wherein z is 0 or 1; R 7 is as defined herein above and preferably is H; n and f are independently an integer

.OH

from 0 to 6, m is 0 or 1 and W is H, CH2Ar or OH and wherein Ar is as 5 defined herein above. Typically suitable phosphate acids and esters include o P- O (OH) 2

I

OH

**O

H

OH

(CH2)n P(OH)2 0 0 wherein m= n 1 to 4; or -OC-CH(CH 3

)-O-P(OH)

2 and pharmaceutically acceptable salts thereof.

The term "heterocyclic ester" as used herein means heterocyclic esters 0 r(CHR 7 )q 1

(CHR

7 y represented by the formula

(CHR

7 )qj wherein

R

7 is as defined herein above, W is an integer of from 1 to 5 preferably W is 1 to 3; q and q' are independently 1 to 4, and q q' are preferably equal to 2, 3, 4, or 5, and Y is CHR 7 NH, NR 7 S, SO or S0 2 Typically suitable heterocyclic esters include 0I0 0 -0 CCH 2 N0 -0 C-CHM. NH -0 C0CH 2 N

S

0 -0CCH 2 -N so 0

-OCCH

2 N 0 0 0 -0 C&CHZN S0 2 0

CCH

2 -N -\N-CH 3 0 and If H 2

N

S.

S..

S

*SS.

0 ~The term 'alkanoate and alkenoate esters" as used herein means straight or branched chain alkanoate or alkenoate groups optionally substituted by a hydroxy or ether moiety or mixtures of such alkanoates or alkenoates.

Preferred alkanoate esters include acetate to decanoate, especially acetate to butanoate. Preferred hydroxy substituted alkanoate ester include Ci to 08 alkanoate substituted one hydroxy moiety or one 01-06 alkoxy group, especially 0CCH 2 0CH 3

-%&CLCH

2 OH and

C-CH(OH)CH

3 Preferred alkenoate esters are the 010-020 alkenoates and include 014 to 018 alkenoates, such as .i- 7 -hexadecenoate.

The term *amino acid ester" as used herein means ax-aminoalkanoyl-oxy groups and includes natural (L)-cx -amino acid ester groups, e.g. the ester of glycine, i.e. OCOCH 2

NH

2 di-and poly peptides esters thereof, unnatural cxamino acid ester groups such as

O-CO-CH(NH

2

)(CH

2 3 C0 2 H, OCOCH(NH2)(CH)2NH 2 0000CH(NH)(CH)3NH2 0 o and a -amino alkanoates represented by the formula

-OCOCH(NR

2 0

R

21

)R

22 wherein

R

20 and

R

2 1 are independently hydrogen or (C1-C8) straight or branched chain alkyl groups or R 20 and

R

21 together with N form a 4, 5 or 6 membered ring optionally substituted with

NR

2 1 or and R 22 is H, CH 3

CH(OH)CH

3 CH2SH,

CH

-CH2 OH I I

CH

2

CONH

2 -(CH2)2CONH 2

CH(CH

3 2

CH(CH

3 2

CH(CH

3 )C2H (CH2) 2

SCH

3 CH2,CO 2 H, (CH2) 2

CO

2 H, (CH2) 4

NH

2

-CH

2 0 6

H

NH

2

CH

2

CH

2

CH

2

,C

NH

1 and pharmaceutically acceptable acid addition salts thereof, or (C 1

-C

8 straight and bianched chain alkyl groups optionally substituted by hydroxyl or NR 20

R

21 Preferred amino acid acids are the natural a -amino acid groups, dipeptides glycyglycine, and a -aminoalkanoates wherein

R

20 and R 2 1 are each

OH

3 The most preferred amino acid esters are Sthose derived from the natural a-amino acids, L-alanine, L-phenylanine, 15 glycine, L-leucine, L-isoleucine and L-valine.

The term Oacid ester" as used herein means those acid esters O

O

11 11 II

II

represented by the formula

O-C-(CR

7

R

7 I)k-C-OH wherein

R

7 is as defined herein above and k is an integer of from 1 to 8. Typically suitable acid esters include oxalic, malonic, succinic, glutaric and adipic acids as well as O

O

branched chain diacids such as -'II'CH-CHCOH branched chain diacids such as ~OC

CHH-OH

The compounds of the present invention thereof exhibit broad spectrum antifungal activity in various in vitro assays against Candida, other yeasts, dematophytes, Asoerillu and opportunistic fungi. The ina vit antifungal activity tests were performed via conventional broth dilution methods in Sabouraud dextrose broth medium and Eagles Minimum Essential Medium ("EMEM") against a large number of fungi. Minimum Inhibitory Concentrations ("MICs") were measured after 24, 48 and 72 hour tests. In many cases, Minimum Fungicidial Concentrations ("MFCS") were measured after 48 and 72 hours.

The term "opportunistic fungi" include Crvtococcus Histonlasma Blst mvce C ddes FusrmMuor Paracccidies Fn W a Sorothrix, Pnemcv Tri saron as shown by invi activity .in an appropriate animal species e.g. mouse, rat or rabbit. The compounds of the inventions are expected to exhibit activity against many genera and species 15 of protoza, bacteria, gram negatives, gram positives, anaerobes, including Legionella, loa Treanema, Gardnerala, Trichomononas and Trvpannom The preferred compounds of formula III wherein

R

11 is convertible in vivo into mono hydroxy-substituted C4 and Cs alkyl groups, 20 exhibited the following in itr antifungal activity in SDB against 37 species of SAseroiljL fl a, furiatus and other species: geometric mean MICs were in the range of 50.05 to <0.81 (mcg/ml) and geometric mean MFCs were in the range of 0.42 to 23.78 mcg/ml.

The more preferred compounds of formula 1II wherein

R

11 is an amino ester group convertible in vivo into a hydroxy-substituted C5 alkyl group exhibited superior antifungal activity as measured by geometric mean MICs and MFCs in various in vitro assays against C. albicans C krusei ~a_.rat E-I tQrBoa T e laidea

C.

nofQrm.an_ and of the dermatophytes, T rubrum. T. mentagrophvtes and T. tonsurans (after 48 or 78 hours) compared to fluconazole as well as superior anti-fungal activity in the following in vivo models: an eriL tflaus and umi[atus (four strains) in a pulmonary immuno- compromised mouse model (PO-1XDX4D) compared to other azoles e.g. itraconazole, and in a Candida Uican (four strains) systemic model with normal and compromised mice (PO-1XDX4D) compared to other azoles, e.g. fluconazole. The preferred compounds of formula IV wherein

R

9 is a -CH(C 2 Hs)CH(R 6 wherein R6 is a group convertible in vivo into OH exhibited superior antifungal activity comparable to the preferred componds of formula V wherein Rio is a hydroxy group such as those listed in Tables Q, R. and S.

The in vivo oral antifungal activity of the compounds of the present invention were compared to azole antifungals, fluconazole in an Asergius pulmonary infection model in mice. The procedure of David Loebenberg et al. entitled "In vitro and In vivo Activity of Sch 42427, The Active 15 Enantiomer of the Antifungal Agent Sch 39304", Antimicrobial Agents and Chemothera (1992), 36. 498-501 was used. The AsDergillus flavus pulmonary model is also described in European Patent Application No.

0539,938AI published on 5 May 1993.

The preferred compounds of formula Ill exhibited superior antifungal in 20 itro activity in SDB against 37 species of Aspergillus with geometric mean MICs of 5 0.05 to 0.81 compared to fluconazole (geometric mean MIC 32 and with geometric mean MFCs of 5 0.42 to >3.78 compared to fluconazole (geometric mean MFC 32).

The Tables Q, R, and S hereinbelow display the superior in vitro antifungal activity of hydroxy derivatives derived from three preferred compounds of formula III compared to fluconazole. Table Q displays such antifungal activity as the percentage of strains of various fungi with MICs 1 mcg/ml for the three preferred compounds of formula III compared to fluconazole. Table R displays the antifungal activity as the percentage of the same strains with MFCs 1 mcg/ml. Table S displays the in vitro MIC 90 values for hydroxy derivatives derived from the three preferred compounds of formula

III

against the same organisms listed in Tables Q and R.

The most preferred compound of formula III where R 5 Me

OCOCH

2

NH

2 Me 5 showed consistently higher serum levels in mice, rats, dogs and monkeys following oral dosing with a methyl cellulose formulation compared to azoles of similiar structure and also exhibited very long serum half lives following O.D. dosing with good tissue distribution. The above listed most preferred compound of formula III is not an inducer of various 10 cytochrome P-450 liver drug metabolizing enzymes after oral administration in an in vivo rat model o TABLE 0 IN VITRO ANTIFUNGAL ACTIVITY FOR SELECTED COMPOUNDS

OF

FORMULA 1111 PERCENTAGE OF STRAINS WITH MICs:! l MCG/ML Me Me Me MMe ORGANISqM Aspe rg ill us S. S

S.

S

15 Candida albicans Candida krusei 20 Candida tropicalis stellatoidea Candida glabrata STR A NS$ 37 26 16 5 9 3 100 100 100 100 22 100 100 100 100 100 100 22 100 100 100 100 100 100 33 100 100

FLZZ-

0 100 0 100 0 0 100 Cryptococcus neoformans De rm atoph yte s

/N"

v] 2- FLZ fluconazole TABLE

R

IN VITRO ANTIFUNGAL ACTIVITY FOR SELECTED COMPOUNDS

OF

FORMULA Jill PERCENTAGE OF STRAINS WITH MFCss 1 MCGIML

MCML)

pJMe 7 Me sc.Me i-w H -wOH MeS /s R-H Me Me

ORGANISM

Aspe rg ill us 15 Candida albicans Candida krusei 20 Candida tropicalis stellatoidea STR A NS 37 26 16 5 9 3 6 50 100 88 100 22 100 67 62 100 94 100 22 100 83 89 100 100 100 22 100 100

FLZZ

0 100 0 100 0 0 0 Candida glabrata Cryptococcus neoformans Derm atophytes 0j N N 1 2. FLZ fluconazole TABLE

S

IN VITRO ANTIFUNGAL ACTIVITY FOR SELECTED COMPOUNDS

OF

FORMULA 1111 MIC 90 Valvesa -Me 2 Me pMe i-OH -OH

O

Me e/S MeR

ORGANISM

Aspergillus Candida 15 albicans Candida krusei 20 Candida tropicalis stellatoid-ea Candida glabrata Cryptococcus neoformans Dermatophytes 1.

STRAI NS 37 26 16 5 9 3 6 .122 .274 .058 .117 28.8 .05 .165 .096 .174 .014 .117 17.1 .007 .101 .112 L"Ir- 29.9 .139 .887 .12 .354 28.8 29.9 .917 29.3 .707 29.4 SV 2. FLZ fluconazole a. Minimum Inhibitory Concentrations for 90% of the strains The preferred compounds of the present invention of formula III or IV are soluble and/or suspendible in an aqueous medium suitable for IV or oral administration and also exhibit superior in vo antifungal activity against a broad range of fungi after oral and parenteral e.g. IV administration In mice, rats, dogs and monkeys. The preferred groups convertible in viv into the corresponding alcohols have a solubility in aqueous medium of at least about 1 to 50 mg/mI, preferably greater than or equal to about 10mg/mI and more preferably about 20 to about 50mg/mi. The preferred esters and ethers of formula IV listed below wherein

R

9 is: 0 N N N-R9

N

IV N

S

R9 F~~M M905 M e 0

-S

me Q0 0 o e 0 M+ 905 Me MeR R §Me M+ 891 Me 0 M+ 817 §Me0 Me 0 0M 0 M+ 861 0 N.Me M e M+ 841 M+ 826

M

MMe M+ 852

M

853 .2M Me

H

OH

M+ 781.7 Rg Me 0 Me A0 H 8NMG M+ 801 §Me M e 'OCH 2

C

6

H

5

OCH

2

C

6

H

M+ 961.4 ,Me Me J-OCH 2

C

6

H

OCH

2

C

6

H

M+ 961.2 *r *4 Rg M e R R 9 Me 0 yl*OO3H.H2.2NMG 0 M+ 881.3 rMe Me 0VOPO,3H2.2NMG 0 M+ 881.3 Me 0

OH

M+ 759.3 Me Me 0 Me 0 M+ 742.4 Me s OH Me -~Me M+ 773.1 §cMe *8*9 (R -oo Me

OH

.2Et 3

N

OH

M+ 873.3 §rMe Me P OH .2NMG

OH

M+ 781.8

M

*0 Me F H.2NMG 0 OH M+ 839 0 M+ 965.7 Me OMe MeS M+ 701.4

R

9 RR 0 Me

NH.CO.CH

2

H

6 M*983 Me meo NNH 2

HCI

M* 758 Sme S 0 Mo

IO

O- OH 2 NMG M 915.4

OH

M+ 915.4 R9 Me Os O Mo M+ 821.5 S Me

HCI

NH

2

HCI

Me Y NH 2

HCI

M+ 801 fl *2NMG

~OH

II OH 0 M+ 929.9 SMe S me

(S

Me .Me 2 NMG 0-MoOH 2 NMG O LO -0 OH

IIOH

M+ 915.8 O M+ 929.2 OH The mass spectral data presented herein as M are parent ions which were determined by Fast Atom Bombardonment (FAB) technique and represent the i.e. {molecular ion+11 peaks.

The more preferred esters listed hereinabove are water soluble (eg and readily convertible in vivo to the corresponding alcohol compounds of the formul V wherein R10 is Me

S

S

OH

Me Me

R

S

Me

OH

Me The most preferred esters are convertible in vivo into alcohols and include those of compounds of formula IV wherein

R

9 is ~Me Me OCOCH 2

O(CH

2

CH

2

O)

3 Me Me W (CHR,)q.

Me -C-CH 2

-NY

Me *2NMG Me 0P0 3

H

2 Me *2NMG Me

OCOCH

2

PO

3

H

2 M e S Me0P0 3

H

2 *2NMG Me 0 CH 3 wherein Y, R 7 and qare as defined herein above and Me S 0 Me

~NH

2

HCI

00 Me

-NH

2 HCl The antifungal compounds of this invention represented by formula I have the R absolute stereochemical configuration at the carbon in the tetrahydrofuran ring bearing the di-halophenyl and liii ,2,4-triazol-1 -ylmethyl moieties, and the CH 2 0Y moiety has the "cis' M stereochemical configuration relative to the 1h, li 2 4 -triazol- 1-ylm ethyl moiety. See the formula

I

hereinbelow.

and Y N NN N wherein R 1 is a straight or branched chain (C4-C5) alkyl group substituted by one or two groups convertible in vivo into hydroxy moities, which preferably exists as a single stereoisomer, but mixtures of stereoisomers are also contemplated as within the scope of this invention.

The compounds of formula I are generically but not specifically disclosed as the "cis" series, type ii, at col. 9 lines 59-68 of Saksena t al. USP 5,039,676 and Example 68 at Col. 5, line 16 to col. 52, line 44.

GENERAL SYNTHETIC

PREPARATIONS

The C4 and C5 hydroxy-substituted alkyl group compounds useful to prepare the compounds of this invention may be prepared by use of the sequence of steps illustrated in the following Schemes I-VI. The compounds of the present inventin are prepared by using the compound 20F of Schemes

II,

and IV-VI in schemes VII to X1B. In Scheme I, compound 3 is readily prepared from commercially available compound 1 according to Examples la, lb and lc.

Compound 4 is prepared by reaction of -diethyl tartarate and molecular sieves in the presence of titanium tetra-isopropoxide (i-PrO) 4 Ti in an aprotic solvent, such as methylene chloride, at a temperature 00 to -350C. See for Example, T. Katsuki, K.B. Sharpless, J. Am. Chem. Soc.. 102, 5974 (1980); 20 and 1J3, 464 (1981). An oxidizing agent, e.g. ert-butylhydroperoxide

("TBHP")

is added to this reaction mixture (step d of Scheme Compound 3 is added and the compound of formula 4 (when L(+)-diethyl tartarate is used) is produced. Reaction of compound 4 with 1H-1,2,4-triazole in the presence of strong base, NaH in an aprotic solvent, such as DMF, at 00-800C provides the diol compound of formula 5. The primary hydroxy group in compound 5 is converted into a leaving group, mesylate or tosylate (compound 6) by reaction of 5 with, for example, mesyl chloride ("MsCI") in an aprotic solvent, methylene chloride in the presence of base, triethylamine ("Et 3 Compound 6 is treated with strong base, sodium hydride (NaH) in an aprotic solvent, DMF at room temperature to give oxirane compound 7.

Reaction of 7 with diethyl malonate in the presence of strong base, sodium hydride in an aprotic solvent, DMSO at 25 0 -75 0 C provides the lactone 8.

Reduction of 8 with a metal hydride, lithium borohydride (LiBH 4 in an alcohol, ethanol (EtOH), provides the triol 9. Conversion of the two primary alcohols of 9 into leaving groups (mesylates or tosylates) by reaction of 9 with excess tosyl chloride in an aprotic solvent, THF, in the presence of base, Et 3 N, provides ditosylate 10. Compound 10 is contacted with strong base, NaH, in an aprotic solvent such as toluene at elevated temperatures of 100 0 -120 0 C to provide a mixture of two tosylates (.jc and trans) which are separated by chromatography to yield to the cis-tosylate 11. Reaction of compound 11 with alcohols HOY in the presence of strong base, such as NaH *in an aprotic solvent, such as DMSO at a temperature of 25 0 -75 0 C provides C4 and C5 hydroxy-substituted alkyl group compounds useful to prepare the 15 compounds of formula i.

Scheme II provides an alternative reaction sequence to obtain compounds of the present invention. Reaction of compound 11 with the commercially available compound 12 in the presence of NaH gives compound .i 13. Hydrolysis of N-acetyl group in 13 is accomplished with a strong base such 20 as NaOH in the presence of n-BuOH to provide compound 14. It should be made clear that instead of N-acetyl group in compound 12, any other base labile groups such as N-formyl, N-benzoyl, etc., can also be used to provide corresponding N-formyl and N-benzoyl derivatives of compound 13. Reaction of 13 with p-chloronitrobenzene in the presence of a hydrochloric acid scavenger such as K 2 C0 3 provides the nitro compound 15. Catalytic reduction of 15 in the presence of a platinum or palladium catalyst yields the amine 16.

Treatment of 16 with phenylchloroformate in the presence of pyridine gives the urethane intermediate 17. Reaction of 17 with hydrazine yields the semicarbazide 18 which is cyclized in the presence of formamidine acetate to furnish the key triazolone 19. Alkylation of 19 according to Examples 19 and provides the compounds of structure 20F including C4 and C5 hydroxysubstituted alkyl group compounds useful to prepare the compounds of formula I wherein

R

1 is defined as hereinabove.

Scheme III provides a stereospecific access to the cis-alcohol 26 and cis-tosylate 11 by application of enzyme chemistry. For Example, reaction of the triol 9 with ethyl acetate in the presence of porcine pancreatic lipase gives a single monoacetate 21. The remaining primary hydroxy group in 21 is protected by an acid labile group such as tetrahydropyranyl group to give a compound such as 22. Hydrolysis of the acetoxy group in 22 is accomplished with a base such a KOH which provides 23. The remaining steps are: (i) tosylation of compound 23 to provide 24; (ii) cyclization of 24 in the presence of NaH to provide 25; (iii) deprotection of THP ether in 25 using an acid catalyst Ssuch as p-toluene sulfonic acid (to give 26) followed by tosylation of the S 15 resulting 26 to furnish the key intermediate 11.

A detailed description of a preferred preparation of key intermediate is disclosed in commonly Sowned Australian patent no. 681659 filed 28 April 1994, which is hereby incorporated by reference.

S S SCHEME

I

0 x1 QAc

OH

a,b

OHI

1 2 OMs

OH

X R OH X R OH X s ~Nf e e6 N

OH

~N N4 *9 0 COOEt

OH

R

XR

Ih

XR

*0

OH~O

N- NX&

N-N

N NkN) 0 m OTs

N~*

X =F or CI fliAgts: NaQAc; Wittig Reaction; KOH; L-DET, TBHP, (i-Pr) 4 Ti; NaH, 1 2 ,4-triazole,DMP: MsCI, Et 3 N, CH 2 01 2 0-5 0 C; NaH,DMF; NaH, diethylmalonate, DMSQ, 50-55 0 C; LiBH 4 EtOH; TsCI, Et 3 N, OMAP,

CH

2 C1 2 -THF NaH, toluene, 100 0 C; (I)Chromatography; (in) NaOY,

DMVSO.

SCHEME 11 H HO-\/z N NCOCH 3 12 (X Cl or F~)

S.

S

0SS 0 06

S

OS

@6

S

S.

@SSS

S

5S S S *5 Ib ~N NH N0 2 *see S:*0

N-N

16 N "JOPh 18 (cont. next page) 17 N SCHEME. 11 CONT'D.

(17 cant.)

NH

NH 2 g IR

NH

NN

Reagents: NaH, DMF; NaOH, fl-BuOH; p-CI-0 6

H

4 N0 2

K

2 00 3 DMSO;

H

2 Pt-C; 0 6

H

5 00001, pyridine,

CH

2

CI

2 Mf

NH

2

NH

2

.H

2 0, H 2 0, dioxane; formamidine acetate, DMF, 8OOCI (h) according to Examples 19 and SCHEME III

OH

OAc b

*O.THP

OAc a

O.THP

O.THP

OH

OTs -wd g- Reggents. Porcine Pancreatic lipase, EtOAc; dihydropyran,

H',

CH

2 01 2 KOH; tosyl chloride, pyridine; NaH, toluene;

H

2 0, methanol, tosyl chloride, pyridine.

SCH-EME

IV

0 M N N H 27 IWi Aq. HBr (ii) SEM.CI H, 0C)N--Nk-SEM -F 0T 2 9

F

29 11F

N

toto (iii)NaH,

DMSO

:F N N AlN- X FN-N EI 3 0X= SEM 0 4. N19

X=

~2OpX R- SCHEME V

(X=F)

(by crystn.) 0- N,12 N 02 Na S

S

S*

55

S

*SS.

S

555

S

NNNH 2 -quant.

1 6F

N-

LN\

\0 N OP h -a \-jH

WN

N

17F H

I

NH

2 18F 0.

-N N

NH

N

19 9F N-

N

I N

I

f m SCHEME

VI

RIX COOR R' 1 Ya No b OH

OH

36 0 R 1 No CR1X CHO 0 p4X

O)CH

2 Ph 37 38

R

4 X is Preferably CH 2 Ph 1 14 RIX is Preferably

CH

3 a.

S

N-NHCHO

YkH

OCH

2 Ph 39

H-N-NHCHO

A R 2

X

OCH

2 Ph SYN isomer 9:1 0

H

1 7F 0

R,

~N-N

N

R, Me M eS

O

R1X=Me pyrrolidine, 24 h; R 4 XX, NaH, DMF; RED-AL, toluene.

H

2 NNHCHO, MeOR; R 2 XM gB r, Et 2 0, -1I0 0 C to r. L, 24 h; 17h of Scheme V and procedure of Example 32d;

H

2 Pd, HCOOH, 80 0

C.

Preparatio Qf POLyQthe Esters Ra(OCHR 7

CHR

7 )tOH LG-(CHR 7

)SCO

2 Na(3) At) Base/THF R8(OCHR 7

CHR

7 )t-O(CH R 7

)S-CO

2

H

a a jDCCD, DMAP', 44

CH

2 01 2 0 0

(CHR

7

)SO(CHR

7

CHR

7 O)tR 8

N-N

6N 1 DCCD Dicyclohexylcarbodiamide DMAP 4-(N,N-Dimethylamino)Pyridine Table for Scheme VII R Me

OH

Me using 20F (X F) A2 1 M. S.

M+

PGOCH

2

CO

2

H

PG Protecting Group,

CH

2 Ph

COCH

2 0H 759.3 I i CH3(OCH 2

CH

2 3 0H ICICH 2

CO

2

H

COCH2O(CH2CH2O)3Me 1-; 43 CH3(OCH 2

CH

2 )QH CICH 2

CO

2 Na -CCH2O(CH 2

CH

2 0)Me 817 CH3(OCH 2

CH)

2 0H CICH 2

CO

2 Na -CCH 2 0(CH 2

CH

2

O)

2 Me 861 CH3(OCH 2

CH

2 3 0H CICH2CO2Na -COCH2(CH 2 CH20) 3 Me '903 HO2C(OCH 2

CH

2 2

CICH

2

CO

2 Na -CCH2O(CH 2

CH

2

O)

2 Me 905

OH

a PEG2000

CICH

2

CO

2 Na -COCH 2 OPEG2000 a PEG5000 CICH2CO 2 Na -COCH2OPEG5000 a A range in MS values were observed which corresponded to the molecular weight range of the PEG2000 and PEG5000 used as starting materials.

CCC.

C. C C C

C.

Phosphate Esters Scheme VillA R- Me Me O 0- 2 N-P-(0CH 2 0 6

H

5 2 46R, S Me R, Me 0 e P(OCH 2 C6HB)2 Tetrazole, t-BuOOH Pd/C,

H

2 AcOH, ETQH 0 M N S0P0 3

H

2 Me 47

N-N

N 2~ 2NMG 47'* 2NMVG SCHEME VIIIB R, R

-M

Me HO{04C_ 7 )LG (49) R, DOOD, DMAP,

CH

2

CI

2 LG Hal Me M 0(CHR)nHa Me 1. AgOP(0CH 2

C

6

H

5 )2 2. ACOH 1O%Pd/C, H 2

ETOH

et *5S* a a 0 11 (CHR1 7 )n-0-P(OH)2 z Table for Scheme VIlA

X=F

47 RI Me

S

me/ 'm P 3

H

2 M.S. .2NMG 781.8 Me

S

.2NMG am0P0 3

H

2 Me Me 0 P(OH)-00 6

H

4 0H 781.7 873.3 47 RI Me

R

~I0

OP(OCH

2

C

6

HS)

2 Me Me me/ P(OCH 2 C6H 5 2 Table for -Scheme

VIIIB

iaa 1 M.S. 961.4 961.2 a a a. a a.

a a M.S.

HOCOCH

2

CI

HOCO(CH

2 4 0H Me

S

mic .2NMG Me/-QMCOCH 2 OPO 3

H

2 839 R 111111111 881.3 HOCO(0H 2 4 0H

S-HOCOCH(.OH)CH

3

\'OCO(CH

2 4

QPO

3

H

2 me .2NMG )dOCOCH 2 4

OP

3

H

2 me Me 0 O P0 3

H

2 Acz 1 1 1 Me .2NMG

OH

3 881.3 853.2 Sheme VIIIC N

N

-0-

AR=

0 AR-N .u L- Me O HO 'CH 2 Oh54 DCCD, DMAP,

CH

2

CI

2 0 _N Me C HOPh 0 EtOH, AcOH a a eo- f (a FR N M e S~ A R-N NH 0 OCH 2 Ph Me 0 56 4 (b) AR-TN> A R-N RN Me y C2OFOH 58 0

Y

N-P(OCH

2 Ph) 2 tetrazole, (ii) t-BuOOH; 10% Pd/C, 58.2NMG

H

2 ETOH, AcOH; 2 NMG Table for _Scheme VIlIC 54a 0-(HO2CCH2C 6

H

4 0CH 2 Ph 0- (H02C(CH 2 2

C

6

H

4 0CH 2 Ph 58 RI 2 NMG 92 .2m 929.9 2 NMG a a.

a.

a a a Scheme IX Preparation of Heterocyclic Ester Me R, S S Hal-(CHR 7 ),COHal OH 62

H

3 C Base H 0 Me 0 N N0(CHR 7 Hal :0 Me a N-N0 N-N r (CHR7)q HN y (CHR7)

HH

0 e Me 110s 0-l N N-G N r N (CHR7)q rl M e KH7)w-- y 0 \,(CHR7I)qI 0 0

N-N

N 61 Table for Scheme IX W61 RI M.S. (M) S*Me 828

OCOCH

2 0 Me Me

S

841

OCOCH

2 -N NMe MeM Me

S

826 MeI COCH 2

Z

SCHEME

X

'SMe

'{OH

Me

S

N -N

(IN)

20F 0

M

N G& -Nf N Q N N S§M Me S >rNH.P 640 9* a p a.

a.

*P.e p. p p a.

a epaa -cS- NN-7-N Me M Me S \Y>I H 3

.C

65 c N- N

(IN\

Reagents: R' COOH, DCCD, DMAP,

CH

2

CI

2 HCI in cijoxane Where

OH

3

C

2

H

5 H; R 2 H, CH 3

CH

2 OH, CH(OH)CH 3

CH

2 SH, CH 2 00NH 2 CH2CH 2 00NH 2

CH(CH

3 2

CH(CH

3

)CH

2

CH

3

CH

2

CH

2 SMe, CH 2

COO-,

CH

2

CH

2 COcy,

(CH

2 4 &t 3

H

2 Cfj 1

H

2 C

OR

3

H

2 C4

Q,

P1i H 2

CH

2

CH

2

CH

2

C<

ri- 11

'NH

R

3 H, Phosphate ester, Sulphate ester; and P N-tert..butoxycarbonyl (N-t-Boc), or N-Carbobenzyloxy (N-Cbz) SCHEME XIA Br R Br 0 (c) (e) a. a a.

a.

a a. a a a 4D *a 7f)(g) 'N 0 76 0 0 O 1 OH 2NG74 1. OCHPh 0 H0

OCH

2 Ph Reagents: 2 2 2 -trichloroethanol; silver dibenzyl phosphate; Zn, HOAc-THF; rMe OC12 (e)20FRI=H

H

2 10% Pd-C; 2eq. N- methyl glucamine.

me SCHEME XIB

CH

3 H0 2 C

I~

(d) 0

HO

2 C I /OCH 2 Ph

OCH

2 Ph *0 OS S 000 0 0* 5 0 0S 0S

SOSS

0 .OtS 0 S. S *5

S.

0 AR-N AN S M g M e 1 0 84 1I*.O0H 2NMG O H Reagents: 2 2 ,2-t ichloroethanol, DCCI), DMAP;(b) N-bromosuccinimide; silver dibenzyl phosphate; Zn, HOAc-THF; 20F, DCCD, DMAP;

H

2 10% Pd-C; 2eq. N- methyl glucamineMe Scheme IV provides an additional reaction sequence to obtain the C4 and C5-hydroxy-substituted compounds useful as intermediates for compounds of the present invention. Compound 27 is prepared from the methyl ether of compound 12 in Scheme II by subjecting the methyl ether of 12 to the reactions of steps a to g of Scheme II. Reaction of compound 27 with aqueous HBr or BBr3 gives phenolic compound 28. Reaction of compound 28 with one equivalent of NaH and subsequent treatment with, for example, 2-(trimethyl)silylethoxymethyl chloride ("SEM-Cl")and DMF at ambient temperatures produces SEM-protected compound 29. Deprotonation of compound 29 with 10 NaH followed by reaction of the so-formed anion with tosylate 11 in DMF or DMSO at elevated temperatures produces compound 30. The nitrogen protecting group of 30, SEM is removed by treatment with, for example, 6NHCI in methanol at ambient temperatures for 3 hr to produce compound 19.

Compound 19 is treated with NaH and DMSO at 20 0 C for 3/4 hr. and thereafter alkylated with RiX to produce compound I. In R1X, R1 is a C3-Cs alkyl group having at least one protected hydroxy moiety, O-SEM and X' is a leaving group, for example, brosylate. Removal of the hydroxy protecting group from compound 31, O-SEM is accomplished by, for example, 6NHCI in methanol to give C4-C 5 hydroxy substituted alkyl group compounds useful to prepare compounds of this invention of formula

I.

Scheme V provides a preferred route for preparation of the intermediates for the compounds of this invention set forth in Scheme II. The sodium salt of compound 31 prepared by reaction of 4 -[4-(4-nitrophenyl)- 1 -piperazinyl]phenol with NaH in anhydrous DMSO at 500-600 C for 30 minutes is reacted with the 2, 4 -diflurophenyl tosylate 11F (compound 11 in Scheme II wherein X=F) for 1 h.

at 500-700 C to provide, after flash silica chromatography or crystallization, compound 15F (compound 15 in Scheme II wherein Reduction of 15F by hydrogenation in the presence of 5% Pd/C in ethanol containing 1NHCI provided amino compound 16F (compound 16 in Scheme II wherein

X=F).

Reaction of 16F with phenylchloroformate in anhydrous pyridine at 0-50C for 2h.

provided phenylcarbamate 17F (compound 17 of Scheme II wherein X=F).

Reaction of 17F with hydrazine hydrate in 1,2-dimethoxyethane at 800C for 4h.

provided the semicarbazide 18F (compound 18 of Scheme II wherein X=F).

Reaction of 18F with formamidine acetate and Et 3 N in 2-methoxyethanol under dry argon in stirred reactor at 80 0 C overnight provided 3H-1,2,4-triazol-3-one 19F (compound 19 in Scheme II wherein Reaction of compound 19(f) with R 1 X in accordance with the procedure of Scheme IV produced the hydroxy-substituted compounds 20F useful to prepare the compounds of 10 formula I.

*'..**Scheme VI provides an alternative, stereoselective route for preparation of the intermediates for preferred compounds of this invention.

Compound 35 S-lactic acid methylester) is contacted with excess 15 pyrrolidine in methylene chloride for 24 hours at room temperature to give amide 36. Reaction of 36 and NaH with for example, benzyl halide in DMF gave 37. Selective reduction of amide 37 with a 3.4M solution of sodium bis(2methoxyethoxy)aluminum hydride ("RED-AI") in toluene at -200C gave aldehyde 38. Reaction of aldehyde 38 with H 2 NNHCHO in methanol gave 39 which was reacted with a Grignard reagent e.g. ethylmagnesium bromide in dry ether at a temperature of -100C to room temperature for 24 hours to give 40 wherein the ratio of the S,S isomer: S,R isomer was 94:6. When the Grigand reaction was done in the presence of 1.2 equivalents of bis(trimethylsilyl)acetamide the SS to SR ratio was 99:1. Compound 40 was reacted with compound 17F of Scheme V in toluene in the presence of DBU (1,8-diazabicyclo[5.4.0]undec-7-ene) for six hours at 800C. Cyclization was effected by raising the temperature to 1000- 1100C and continuing, to maintain this temperature overnight. After purification via TLC, 20F was obtained. Treatment of 20F with hydrogen and palladium black in methanol containing formic acid heated to 600C gave the crude product which was isolated and purified (via TLC) to give compound 20F i.e. the compound of formula V wherein Me

S

R

10 and X F, Mt 701

OH

Me The reaction of the Grignard reagent on the propanimine 39 produces 40 wherein the absolute stereochemistry induced at the new chiral center in 40 is substantially the same S) as that at the chiral carbon in 39.

By the term "substantially the same" as used herein is meant the ratio of S:S to S:R (in 40) is greater than 9:1, preferably is greater than 15:1 and most 10 preferably is at least 99:1.

The mass spectral data presented herein as M+ are parent ions which were determined by Fast Atom Bombardonment (FAB) technique and Srepresent the i.e. {molecular ion+1) peaks.

5 We consider that Schemes I-VI set forth the preferred S 15 processes to prepare the C 4 alcohol intermediates for the compounds of this invention. Other processes for preparing the alcohol intermediates for the compounds of this invention are disclosed in commonly-assigned U.S. Patent No. 5,616,777 and US Patent No. 5,625,064; and these two patents are hereby incorporated by reference.

Scheme VII provides a general method for preparation of the polyether esters of the present invention. The alcoholate of alcohol ether 42 e.g. CH3(OCH 2

CH

2 3 0H 42 wherein

R

7 H and t 3, was prepared by reaction, of 42 with excess strong base e.g. NaH in an anhydrous ether e.g. THF at ice bath temperatures. The so-formed reaction mixture was stirred for several hours 2 or more and the sodium salt of acid 43 e.g. sodium salt of chloroacetic acid (43 wherein LG is a halogen(Hal) =CI, R 7 =H and s=1) was added thereto. The so-formed reaction mixture was stirred at ice-bath temperatures and stirring was continued as temperature was allowed to warm to room temperature. Water was carefully added to the reaction mixture and the polyether acid 44 was separated and purified by conventional techniques.

To a solution of 44 in CH 2

CI

2 was added 1.3-1.5 equivalents of the base 4 -(N,N-dimethylamino)pyridine ("DMAP") and 20F wherein Me

S

M e Me OH OH The temperature of the so formed reaction mixture was lowered by use of an ice bath and 1.3-1.5 equivalents of 10 dicyclohexylcarbodiimide ("DCCD") was added thereto. The so-formed reaction mixture was continuously stirred as the temperature was allowed to warm to room temperature. The dicyclohexyl urea precipitate was removed and the crude product isolated by conventional techniques. The so formed residue was purified by chromatography on silica gel to provide the pure compound [M H]+ S 15 906 by FAB. By the appropriate substitution of different starting materials 42 and 43 the compounds 45 listed in Table for Scheme VII were prepared. The .MS values for products listed under 45 in the Table for Scheme VII were measured by Fast Atom Bombardment Schemes VIII A-C illustrate the generalised methods for preparing phosphate esters of this invention. Scheme VIllA provides a method for preparation of phosphate esters of formula IV wherein Re is O9

O

11 II 0 f- (CH2R7)n-

P(OW)

2 and z m=n 0. Compound 20F of Scheme II in methylene chloride at room temperature was reacted with equivalents of N,N-diisopropyl-dibenzylphosphoramide, and 3 equivalents of a $r a base such as tetrazole, followed by 1.5 equivalents of tert-butyl peroxide (3M in iso-octane) for several hours. The progress of the reaction was followed by TLC methanol:EtOAc v:v) on silica gel. The crude product in EtOAc was washed with sodium thiosulfate and purified using standard techniques to provide the dibenzylphosphate ester 46. The dibenzyl ester groups of 46 were removed to give 47 by treatment of 46 dissolved in equal volumes of ethanol and glacial acetic acid in the presence of 10% Pd/C under a hydrogen atmosphere at room temperature in a stirred reactor overnight. The reaction was continued until no starting material was found by TLC (or NMR). The catatyst was removed by filtration and the crude phosphate ester 47 was purified by standard techniques.

Treatment of 47 in methanol at room temperature with two equivalents of base e.g. NMG (or Et 3 N) provided 47 2NMG. The compounds 46 and 47 prepared in accordance with Scheme VIIIA are listed in the Table for Scheme

VIIA.

Scheme VIIIB illustrates preparation of phosphate esters of O

O

11 11 formula IV wherein R 6 Z z (H2 )n (OW)ndn z =m=1 and n o. Compound 20F dissolved in methylene chloride was treated with 1.3 equivalents of DMAD 1.3 equivalents of DCCD and 1.3 equivalents of the acid O4 11 HO-U -C-(CH2R7)n-LG 49 of tho fnrmla Z HO2C(CH2)4Br, z 1, n 4,

R

7 H and the leaving group LG is a Hal Br. The reaction was stirred at room temperature until no starting material was found by TLC purification of the crude product gave bromide 50, a white solid wherein R Me A Me/ ,0 2

(CH

2 4 Br SThe bromide 50 in benzene was heated at 80 0

C

overnight with 1.5 equivalents of silver dibenzylphosphate (available from Sigma Chemical Co., St. Louis). The reaction mixture was cooled and washed with aqueous base, K2CO3. The crude product was separated and purified by silica gel column chromatography to give the dibenzyl phosphate ester 51.

Treatment of 51 in ethanol/glacial acetic acid with 10% Pd/C under a hydrogen atmosphere overnight at room temperature gave phosphate ester 52.- Treatment of 52 in methanol with two equivalents of base e.g. NMG (or Et 3

N)

gave 52 2NMG.

Scheme VIIC provides an alternative procedure for preparation of 10 phosphate esters of formula IV wherein

R

6 is as defined above for Scheme

VIIIB

and z 1 and n 1. The benzyl ether of methyl acetate 53 in methanol-water .i and excess base e.g. K2CO3 were stirred overnight at room temperature to give the benzyl ether 54. Reaction of a solution of 20F and 54 in methylene chloride with a 1.3 1.5 equivalents of DCCD and DMAP at room temperature overnight 15 gave ester 55. The benzyl ether group of 55 was removed by treatment with excess 10% Pd/C in ethanol-glacial acid under a hydrogen atmosphere at room temperature overnight. Purification of the crude product gave 56. Treatment of 56 with 1.5 equivalents of N N-diisopropyldibenzylphosphoramide and 3 equivalents of tetrazole and followed by 1.5 equivalents of tert-butyl peroxide in accordance with the procedure of Scheme VIIIB gave dibenzyl ester 57.

Removal of the dibenzyl groups with 10% Pd/C in ethanol-glacial acetic acid under hydrogen atmosphere gave (as described hereinabove) phosphate ester 58. Treatment of 58 with two equivalents of base, e.g. NMG, gave 58 2NMG.

Additional phosphate esters of this invention of the formula O C(CHR)n

(CHR

7 )f

O

I

II

(O)m--P(OW) 2 wherein z=m=1, n= 1 or 2, f=0 or 1, and W=H, such as listed in the Table for Scheme VIIIC are prepared from p.

p p p p

S..

p.

S. p compound 20F wherein R 1 is the same as that used in Scheme VIIIA by substituting equivalent amounts of starting materials 54a and 54b for compound 54 and thereafter following the procedures shown in Scheme

VIIIC.

Scheme IX illustrates the preparation of heterocyclic esters of the RMe Rl Me M e

-OH

present invention. Compound 20F, wherein Me dissolved in methylene chloride is reacted with compound 62 in the (Hal=Br or CI,

CI-CH

2 -COCI) in presence of a base such as pyridine at a temperature of 0°-5C for four hours. The reaction was placed in a refrigerator overnight.

Additional compound 62 and base could be added, if necessary, and the 10 reaction continued until no 20F is present by TLC. Purification of the crude product by column chromatography on silica gel gave pure 59 Hal=CI).

Reaction of 59 with excess of the nitrogen heterocyclic compound 60 Y=NH,

R

7 =H and q=4) at a temperature of 500-600C for 1 hour produced 61.

Substitution of nitrogen heterocyclic compound 60 with a five and six membered compounds, e.g. morpholine, N-methylpiperidine provided the compounds listed in Table below Scheme

IX.

Scheme X illustrates preparation of the amino acid esters of this invention. Compound 20F is contacted with excess N-(-butoxy carbonyl a -amino acid or a -amino alkanoate in the presence of DCCD and DMAP in an aprotic solvent such as CH 2 C12 at 00 to 250C. The reaction is followed by TLC and additional a -amino acid and DCCD are added, if necessary, to insure the starting material 20F is completely converted into amino acid ester derivative 64. Compound 64 is treated HCI in dioxane to provide the a-amino acid ester as the acid addition salt 65. Purification of the crude products is accomplished by standard techniques. When carbobenzoxy is used as the protecting group, hydrogen over palladium black is used to remove the protecting group in step b.

Other protecting groups may be used such as those disclosed in "Protective Groups in Organic Synthesis" by T.W. Green and P.G.M. Wuts. John Wiley and Sons 1991 NY at pages 97-98 or 389-394.

Schemes XIA and XIB illustrate the praparation of additional phosphate esters of this invention of the -0 C(CHR)nQ I

I

(C

HR

O

formula 0

P(OW)

2 wherein z=f=m=l, n=0 or 1, and W=H.

In Scheme XIA, the benzoyl bromide 70 is treated with 2, 2, 2trichloroethanol to produce the corresponding trichloroethyl ester. Treatment of the trichloroethyl ester with excess silver dibenzyl phosphate under conditions 10 similar to those used in Scheme VIIIC converted the benzyl bromide into a dibenzyl phosphate ester Removal of the trichloroethyl ester group was accomplished by use of zinc in acetic acid THF to give the dibenzyl phosphate ester 72. Treatment of 72 with thionyl chloride gave the corresponding acid chloride which was contacted with a solution of 20F in methylene chloride under 15 conditions of step one in Scheme VIIIC to give ester 74. Removal of the dibenzyl ester groups of 74 with 10% Pd/C under a hydrogen atmosphere as described in Scheme VIIIA produced the corresponding phosphate ester which was treated with two equivalents of base e.g. NMG to provide 76. In Scheme XIB, the 2 -methylphenylacetic acid 80 was esterified with 2, 2, 2-trichloroethanol and the so-formed ester was converted into corresponding the benzyl bromide by treatment with N-bromosuccinimide. The treatment of the benzyl bromide with excess silver dibenzyl phosphate under conditions of Scheme

VIIIC

provided the corresponding dibenzyl phosphate ester. Removal of the trichloroethyl ester group was accomplished with zinc in acetic acid-THF to give dibenzyl phosphate 82. Treatment of a solution of 82 and 20F (wherein

R

1 is the same as in Scheme VIIIA) with DCCD and DMAP provided the corresponding phosphate ester. Treatment of the phosphate ester with two equivalents of NMG gave compound 84.

The alkanoate and alkenoate esters of 20F are conveniently prepared by standard synthetic techniques, (for example, by reaction of the anhydride or acid halide of the alkanoic acid or alkenoic acid in tghe presence of base e.g, pyridine) produced the alkanoate or alkenoates of the compounds of formula I.

The sulfate esters may be prepared by reaction of the alcohol S: 10 compounds of formulas I to IV with sulfur trioxide in the presence of excess pryridine at temperatures of 70°-90°C for at least 2 hours in accordance with the procedure of R.M. Moriarty et. al. Tetrahedron Letters, Vol. 35, No. 44, p 8103-8106 (1994).

The corresponding alcoholic compounds of formula I may also be 15 prepared by reaction of compound 11 with alcohols of formula HOY in the presence of a strong base, NaH in an aprotic solvent, such as DMSO.

H Ts X R

OY

J I yO O -OY

-N

X N-N

N

-N

N N (R)-"Tosylate" Series See Example wherein X F or Cl 0

R

N N and R 1 a (C4-05) alkyl group substituted by one or two hydroxy moieties.

Compounds of this invention represented by formula I exhibit broad spectrum antifungal activity, in conventional antifungal screening tests, against human and animal pathogens, such as the following: Aspergillus, Blastomyces, Candida, Cryptococcus, Coccidioides, Epidermophyton, Fonsecaea, Fusarium, Mucor, Saccharomyces, Torulopsis, Trichophyton, Trichosporon, Sporothrix and Pneumocysitis.

The preferred compounds of formula IV exhibit topical, oral and parenteral antifungal activity in in vivo tests in animals and such activity is unexpectedly better than that of existing antifungal agents e.g. itraconazole and 10 fluconazole as well as that of the azole compounds specifically disclosed by Saksena eta. in USP 5,039,676 and International Publication No.

WO 93/09114.

The antifungal compounds of formula I and pharmaceutical compositons of this invention are expected to exhibit anti-allergic, anti- 15 inflammatory and immunomodulating activities, broad spectrum antiinfective activity, antibacterial, anti-protozoal and antihelminthic activities.

~The present invention also provides a composition for treating or preventing fungal infections comprising an antifungally effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent therefor.

The pharmaceutical compositions of the present invention may also contain a fungicidally effective amount of other antifungal compounds such as cell wall active compound. The term "cell wall active compound", as used herein, means any compound that interferes with the fungal cell wall and includes, but is not limited to, compounds such as papulacandins, echinocandins, and aculeacins as well as fungal cell wall inhibitors such as nikkomycins, e.g, nikkomycin K and others which are described in USP 5,006,513 which is hereby incorporated by reference.

The pharmaceutically acceptable salts of the compounds of the present invention include pharmaceutically acceptable acid and base addition salts.

The preferred pharmaceutically acceptable acid addition salts are nontoxic acid addition salts formed by adding to the compounds of the present invention about a calculated amount of a mineral acid, such as HCI, HBr,

H

2 S0 4 HN3O or H 3

PO

4 or of an organic acid, such as an alkyl or arylsulfonic acid such as methanesulfonic, isithionic, para- toluenesulfonic, naphthylsulfonic and the like.

10 The pharmaceutically acceptable bases found suitable for use in the present invention are those which form pharmaceutically acceptable salts of the acidic pharmaceutically acceptable esters of the antifungal compounds of formulas I, II, Ill or IV and include suitable organic and inorganic bases.

Suitable organic bases include primary, secondary and tertiary alkyl amines, 15 alkanolamines, aromatic amines, alkylaromatic amines and cyclic amines.

.:Exemplary organic amines include the pharmaceutically acceptable bases t* selected form chloroprocaine, procaine, piperazine, glucamine,

N-

methylglucamine, N-N-dimethyl glucamine ethylendediamine, diethanolamine, diisopropylamine, diethylamine, N-benzylenediamine, diethanolamine, diisopropylamine, diethylamine, N-benzyl-2-phenylethylamine,

N-

n'dibenzylethylenediamine, choline, clemizole, triethylamine

("ET

3 tris(hydroxymethyl)aminomethane, or D-glucosamine. The preferred organic bases include N-methyl glucamine diethanolamine, and tris(hydroxymethyl) aminomethane Use of two equivalents of NMG in this invention is more preferred. The suitable inorganic bases also include alkali metal hydroxides such as sodium hydroxide.

The pharmaceutical compositions of the present invention may be adapted for any mode of administration for oral, parenteral, SC, IM. IV and IP, topical or vaginal administration or by inhalation (orally or intranasally) Such compositions are formulated by combining the compound of formula I or an equivalent amount of a pharmaceutically acceptable salt of compound I with an suitable, inert, pharmaceutically acceptable carrier or diluent.

Examples of suitable compositions include solid or liquid compositions for oral administration such as tablets, capsules, pills, powders, granules, solutions, suppositories, troches, lozenges, suspensions or emulsions. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders or tablet disintegrating agents; it can also be an encapsulating material.

In powders, the carrier is a finely divided solid which is in admixture with the finely divided active compound. In the tablet, the active compound is mixed with :00 carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.

Topical dosage forms may be prepared according to procedures 15 well known in the art, and may contain a variety of ingredients, excipients and additives. The formulations for topical use include ointments, creams, lotions, 000 powders, aerosols, pessaries and sprays.

For preparing suppositories, a low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted, and the active ingredients are dispersed homogeneously therein as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool and thereby solidify.

Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection. Liquid preparations can also be formulated in solution with an appropriate amount of a hydroxypropyl a- P or -y-cyclodextrin having 2 to 11 hydroxypropyl groups per molecule of cyclodextrin, polyethylene glycol, PEG-200 or propylene glycol, which solutions may also contain water. Aqueous solutions suitable for oral use can be prepared by adding the active component in water and adding suitable colorants, flavors, stabilizing, sweetening, solubilizing and thickening agents as desired. Aqueous suspensions suitable for oral use can be made by dispersing the active component in finely divided form in water. A particularly preferred aqueous pharmaceutical composition may be prepared from the compounds of'formulas

I

to IV together with hydroxypropyl-p-cyclodextrin in water. The use of derivatives of P- and y-cyclodextrins, for example, hydroxpropyl--cyclodextrin are disclosed by N. Bodor USP 4,983,586, Pitha USP 4,727.064 and Janssen Pharmaceutical Australian Patent No. 565966.

10 The pharmaceutical compositions of the present invention may be prepared by admixing the pharmaceutically acceptable carrier, a S hydroxypropyl-p-cyclodextrin in water, and adding thereto an antifungally effective amount of a drug of the present invention. The solution so formed is filtered, and optionally, the water may be removed by well known methods, e.g., rotatory evaporation or lyophilization. The formation of the solution may take place at a temperature of about 150 to 350C. The water is normally sterilized water and may also contain pharmaceutically acceptable salts and buffers, e.g., "0 phosphate or citrate as well as preservatives. The molar ratio of the antifungal compound of formula I to hydroxpropyl-p-cyclodextrin is about 1:1 to 1:80, 20 preferably 1:1 to 1:2. Normally the hydroxypropyl-p-cyclodextrin is present in molar excess.

Also included are solid form preparations which are intended to be converted, shortly before use, into liquid form preparations for either oral or parenteral administration. The solid form preparations intended to be converted to liquid form may contain, in addition, to the active materials, such as compounds of this invention, and optionally a cell wall active compound, especially a fungal cell wall inhibitor, a nikkomycin, flavorants, colorants, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents and the like. The solvent utilized for preparing the liquid form preparations may be water, isotonic water, ethanol, glycerin, polyethylene glycols, propylene glycol, and the like, as well as mixtures thereof.

Parenteral forms to be injected intravenously, intramuscularly, or subcutaneously are usually in the form of a sterile solution, and may contain salts or glucose to make the solution isotonic.

The topical dosage for humans for antifungal use in the form of a pharmaceutical formulation comprising a compound of formula I (usually in the concentration in the range from about 0.1% to about 20% preferably from about to about 10% by weight) together with a non-toxic, pharmaceutically 10 acceptable topical carrier, is applied daily to the affected skin until the condition has improved.

In general, the oral dosage for humans for antifungal use ranges S. from about 1 mg per kilogram of body weight to about 30 mg per kilogram of body weight per day, in single or divided doses, with about 1 mg per kilogram of body weight to about 20 mg per kilogram of body weight per day being preferred and the dose of about 1 mg per kilogram of body weight to about ooo° 0 *mg per kilogram of body weight per day being most preferred.

In general, the parenteral dosage for humans for antifungal use ranges from about 0.25 mg per kilogram of body weight per day to about 20 mg kilogram of body weight per day, in single or divided doses, with about 0.5 to about 10 mg per kilogram of body weight per day being preferred.

The exact amount, frequency and period of administration of the compounds of the present invention for antifungal use will vary, of course, depending upon the sex, age and medical condition of the patent as well as the severity of the infection as determined by the attending clinician.

GENERAL EXPERIMENTAL The compounds of this invention are prepared in accordance with Schemes 1-IX hereinabove and the following Examples using commercially available starting materials.

F O F O CI OAc NaOAc I'F"

F

i: EXAMPLE la 2-Acetyloxy-1-( 2 4 -difluoroDhenvl)ethanone Add 191 g of 2 -chloro-2',4'-difluoroacetophenone (Aldrich Chemical Co.) to a mixture of 246 g of sodium acetate, 3 g of Nal, and 3 L of DMF. Stir the mixture at 200C for 18 hr. then concentrate it to 1 L. Pour the residue into 6 L of cold dilute aqueous HCI and extract with EtOAc. Wash the 15 extract with brine, dry it over anhydrous Na 2

SO

4 filter the so-formed mixture, and evaporate the filtrate to leave a residue. Chromatograph the residue on silica gel, eluting with CH2CI-2-hexane to obtain 198 g of the title compound.

F O

F

OAc OAc MePh3PBr Na-HMDS F THF

F

F

EXAMPLE lb 1-[ 2 2 .4-Difluorophenyl)1-2-DropenoI acetate Suspend 131 g of MePh 3 PBr in 270 mL of mechanically-stirred, dry THF at 20 0 C. Add 393 mL of 1M NaN(Me 3 Si) 2 in THF, slowly at first, then rapidly over 5 min. while applying just enough ice cooling to maintain the temperature at 23°C. Stir the so-formed mixture for 1 hr at 200-24 0 C, cool it to 70 0 C, and stir it another 1/2 hr. Then add thereto a solution of 65.5 g of the product of Example la in 140 mL of dry THF, at a rate slow enough to keep the temperature below -700C. Continue to stir the so-formed reaction mixture in the cold bath overnight during which the temperature rises to 200C. Add 50 mL of EtOAc to the so-formed suspension, and then add 3 L of hexane. Allow the soformed mixture to stand for -15 min., and suction-filter to remove Ph 3 PO. While the filter cake is still damp, transfer it to a beaker. Triturate the cake thoroughly with 1/2 L of hexane and suction-filter again to remove the remainder of product.

10 Wash the combined hexane filtrates with 2 x 1 L of a 1:1 MeOH-water, and then with brine. Dry the organic layer over MgSO 4 filter and evaporate the filtrate to leave a red oil. Add 1.5 L of hexane and suction-filter through a Celite pad to leave a clear yellow solution. Chromatograph the yellow oil on silica gel, eluting with 1/2 L of hexane, then 1L of 15:1 hexane-EtOAc. Combine the 15 homogeneous fractions to yield 38.6 g of the title compound as an oil.

.F OAc F OH

KOH

F

EXAMPLE 1c 2 2 .4-DifluoroDhenyl)-2-DroDenol.

Dissolve 40 g of the product of Example lb in 400 mL of dioxane.

Add a solution of 18 g of 85% KOH in 315 mL of water. Stir the so-formed mixture vigorously for 1 hr, and then pour the mixture into 1 L of Et 2 0. Separate the aqueous layer and extract it with 250 mL of Et 2 0. Combine the organic extracts, and wash them with water and then brine. Dry the organic extract over anhydrous K 2

CO

3 and add 10 g of charcoal thereto. Filter, and evaporate the filtrate to leave 31.3 g of the title compound as a straw-colored oil.

EXAMPLE id S)-(-)-[2-r2-(24-DifluoroDhenyl)loxiranyllmethanol Add 33g of activated 3A molecular sieve powder to a solution of 13g of L-(+)-diethyl tartarate in 2.3L of CH 2

CI

2 and cool the so-formed mixture to -50C. Add a solution of 15.4 mL of titanium letra-isopropoxide in 100 mL of

CH

2

CI

2 over 2-3 minutes and then cool the so-formed mixture to -22°C. Add 109.5 mL of a 5.5 M solution of ieft-butylhydroperoxide in 2,2,4-trimethylpentane over 4-6 minutes, and cool the so-formed mixture to -250C. Stir the mixture at -250C for 25 minutes and then add a solution of 40g of 2-(2,4difluorophenyl)-3-propenol of Example 1c in 100 mL of CH2CI2 over 3-4 minutes. Stir the so-formed mixture at -270C for 4 1/2 hour. Add 102 mL of aqueous sodium hydroxide saturated with NaCI and stir the so-formed mixture while warming to +10°C over a 1/2 hour period. Add thereto 100 g of anhydrous MgSO 4 and 33g of Celite, and stir 1/2 hour at +10°C. Suction-filter the mixture, 15 wash the so-formed filter cake with 1.2 L of diethyl ether (Et20) and then 1.5L of toluene, and dry the combined organic layers over anhydrous MgSO 4 Filter the organic layer, and evaporate the filtrate in vacuo to form a residue. Dissolve the residue in 1L of Et 2 0 and suction-filter the mixture to remove insolubles.

Suction-filter the filtrate through 100g of silica gel, and wash the pad with 200 mL of fresh Et20. Evaporate the filtrate in vacuo to give 41g of the crude title compound as a yellowish oil, .36.70 MeOH); PMR (CDC 3 6 7.40(m,1H), 6.85(m, 2H), 3.95(m,2H), 3.31(d,1H), 2.84 1.91(m,1H, deuterium exchangeable).

EXAMPLE 2 4 -DifluoroDhenvy)oxirnvllmethanol Follow the procedure of Example id, except substitute an equivalent amount of diethyl tartarate in place of diethyl tartarate to give the crude title compound, [a 33.90 MeOH).

Purify a portion of the crude compound by silica gel chromatography to obtain a sample homogeneous by TLC, a 40.00 MeOH) EXAMPLE 3 (R)-(-)-2-(24-Difluorophenvl)I-3-(1.24-triazol-1-l)-1 .2-ropanediol Dissolve 8.91g of 1H-1,2,4-triazole in 150 mL of anhydrous

DMF

and cool so-formed mixture to 0-50C. Add 2.81g of sodium hydride (60% oil dispersion) and stir the so-formed mixture 30 minutes at room temperature. Add thereto 10.9 g of the product of Example id. Stir the so-formed reaction mixture for 2 hours at 60-700C. Cool the mixture to room temperature, add thereto 10 ml of H 2 0 and evaporate it in acuo to give a residue. Dissolve the residue in 100 mL of H 2 0 and 900 ml of ethyl acetate (EtOAc). Extract the H 2 0 layer with another 250 mL of EtOAc. Wash the combined EtOAc extracts with 100 mL of brine. Dry the EtOAc extracts over anhydrous MgSO 4 and evaporate. Triturate the so-formed oily residue with 10 mL of CH 2

CI

2 and add 100 mL of Et20. Stir the CH2CI 2 -Et 2 0 mixture for 1 hour at room temperature. Filter to give 11.2g of the title compound, [a1 70.7 MeOH), mass spectrum

(FAB):

256 Recrystallize 1.0g of the filtered product from 5 mL of CH 3 CN to give 0.83g of the title compound, m.p. 99-100oC; a 71.50 MeOH); elemental analysis: £aLcuaiI for C11H11F2N 3 0 2 1/2CH 3 CN; 52.27C, 4.57H, 17.78N, 13.78F; Found 52.26C, 4.58H, 17.54N, 13.78F; PMR(DMSO) 8 8.25 7.66(s, 7.33, 7.09(t, 6.90(t, 5.72(s, 5.05(t, 4 .53(s,2), 3.61 EXAMPLE 4 4-DifluorohenvlI-.-(1 .24-ripzoi -1 2 roonedbl Follow the procedure of Example 3, except substitute an equivalent quantity of the product of Example 2 in place of the product of Example 1ito give the title compound; MP. 95-101 OC; IaD+ 70.0' MeOH). The PMR and Mass spectra were consistent with the structure of the title compound.

EXAMPLE 2 -(2.4-Difluoroohenvl).3.(i riazol-i -yb)-.2rol~nediol- 1ethpnesu Ifonate 15 Suspend 10.9 g of the powdered product of Example 3 in 150 mL of CH 2

CI

2 Add thereto 8.95 mL of triethylamine and cool to the so-formed mixture 0-50C. Add 3.64 mL of methanesulfonyl chloride in 20 ml of CH 2

CI

2 over 10 min. Stir the so-formed mixture for 1 hour at room temperature. Cool it to 0-5 0 C, extract with 100 mL of cold (0-.5 0 C) 5% KH 2

PO

4 followed by 100 mL of cold (0-5 0 C) H 2 0, followed by 50 mL of brine. Dry the separated organic layer over anhydrous MgSO 4 and evaporate to obtain 13.7 g of the title 4.65(d,l), 4.46(m,2), 3.05(s,3).

EXAMAPLE 6 2 -(2.4-Diflu )orohenyjjj3.i'i 24-triazol-i v) 2ooaeil1 Metansuf onat Follow the procedure of Example 5, except substitute an equivalent quantity of the product of Example 4 in place of the product of Example 3 to give the title compound. The PMR is consistent with the structure of the title compound.

EXAMPLE 7 Rx)-1- 2 -r 2 -r2.4-Diflo rophenvloxiranvlmethyl .2.4-triazole Dissolve 13.7g of the product of Example 5 in 200 mL of anhydrous DMF and cool the so-formed solution to 10-150C. Add thereto 1.71g of sodium hydride (60% oil dispersion) and stir the so-formed reaction mixture at room temperature for 90 minutes. Concentrate in vacuo to 50 mL. Add thereto 10 200 mL of cold H 2 0 (0-50C) and extract with 3 x200 mL portions of EtOAc.

Wash the combined EtOAc extracts with 100 mL of brine. Dry the EtOAc extracts over anhydrous MgSO 4 and evaporate it to give 10.8 g of a residue. Apply the residue in CH 2

CI

2 to a column of 400 g of MPLC grade silican gel previously prepared by slurry packing with CH 2

CI

2 containing 1 mL of Et 3 N per liter. Elute 15 with 1 liter, each of 25, 50 and 75% EtOAc; CH 2

CI

2 followed by 2 liters of EtOAc. Combine the fractions to give 6.92g of the title compound. Mass spectrum (FAB): 238 PMR (CDCI 3 5 7.97(s,1), 7.77(s,1), 7.07(m,1), 6.73(m,2); 4.73(d,1), 4.41(d,1), 2.84(d,1), 2.78(d,1).

EXAMPLE 8 -12-r2-( 2 .4-difluoroDhenvl)loxiranvlmethvll- 1.2.4-triazole Follow the procedure of Example 7, except substitute an equivalent amount of the product of Example 6 in place of the product of Example 5 to give the title compound. [PMR is consistent with the structure of the title compound].

EXA MPLE 9 Ethy(SRcjs) an (S-trans)-5-(2.4-Dif IuorophenviL).2-o-xo-5-rU1 H-i 2 _A-trip2znOL_ l-vflmnethyULterahydro-3.furancprbowlate Dissolve 9.35 mL of diethyl malonate in 70 mL of anhydrous DMS0. Add 2.24g of sodium hydride (60% oil dispersion) in 2 portions and stir the so-formed reaction mixture at room temperature for 1 hour. Add 6.65 g of the product of Example 7 and stir 18 hours at 50-55 0 C. Cool to room temperature and pour the reaction mixture into a well-stirred mixture of 500 mL of KH 2

PO

4 500 mL of brine, and 1 liter of EtOAc. Separate and extract the H 2 0 layer with another 300 mL of EtOAc. Wash the combined EtOAc extracts with :.500 mL of brine, Dry the EtOAc extracts over anhydrous 'MgSO 4 and evaporate to give an oil. Apply the oil with CH 2

CI

2 to a column of 400 g MPLC grade silica gel prepared with hexane. Elute with 500 mL of hexane, followed by 2 liters of 50% EtOAc: hexane (vlv), followed by 2 liters of EtOAc. Combine fractions to give 8.66g of the title compound. Mass spectrum (FAB): 352[M+H]+,

PMR

(CDCI

3 d 8.08(s,2), 7.91 7.71 7.42(m, 7.13(m, 7.85(m,2), 4.60(m,4), 4.1 3.49(t, 3.14(t, 3.89(m,4), 1.1 8(m,6).

EXAMLQ1 and S-rans)-(.4-Difluororhenv)2oxo5(l H-i .2.4-triazol.1 ylm thyIetrhro~-. f' Waarbxlae Follow the procedure of Example 9, except substitute an equivalent amount of the product of Example 8 in place of the product of Example 7 to give the title compound. [PMR and mass spectra are consistent with the structure of the title compound].

EXAMPLE 11 2 4 -Difluoro henyl)-2-hydroxvmethyl-5-1 H-(1.2,4-triazol-1 -vl)1-1 4- Dentanediol Dissolve 8.5 g of the product of Example 9 in 125 mL of EtOH and add 2.15 g of LCI. Cool the stirred mixture to 0°C and add 1.92 g of NaBH 4 in portions. Stir the mixture for 18 hr without further cooling. Add 125 mL of MeOH and 10 mL of H 2 0 to the mixture and stir for 4 hr. Evaporate the mixture to dryness and extract the precipitate with warm EtOH. Evaporate the extract to dryness, add 200 mL of THF to the residue, and sonicate the stirred mixture for 10 15 min. Filter the mixture and evaporate the filtrate. Chromatograph the residue on silica gel, eluting with CH2CI2-MeOH-NH 4 0H (95:5:1) v/v/v) to obtain 3.9 g of the title compound. Mass spectrum (FAB): 314 PMR (DMSO) 8.25(s,1), 7.69(s,1), 7.35(m,1), 7.13(m,1), 6.94(m,1), 6.27(s,1), 5.16(t,1), 4.44(m,4), 3.39(m,1), 3.20(m,1), 3.05(t,2), 2.11(m,1), 1.52(m,1).

EXAMPLE 12 (S)-(+)-4-(2.4-DifluoroDhenyl)-2-hydroxymethyl-5-r1 H-1 .2.4-triazolvl)1-1.4- Dentanediol 5 F o llo w th e procedure of Example 11, except substitute an

S

equivalent amount of the product of Example 10 in place of the product of Example 9 to give the title compound. Chromatograph a portion of the crude product on silica gel eluting with CH2CI2-MeOH-NH 4 0H to give a product homogeneous by TLC. Dissolve the material in H 2 0 and filter, and lyophilize the filtrate to give the title compound. 1 54.50 MeOH) EXAMPLE13 4 2 4 -Difluorophenyl}.2.r(4methyI hnyI-I)..uf n nvwmthylsr

H

(1 .2.4-triazolyfll-1 .4-1Dentanediol-1 4 -methLbenzene)s ulfonpate Dissolve 4.4g of the product of Example 11 in 50 mL of CH 2

CI

2 THE vlv). Add 4.7 mL of Et 3 N and 180 mg of N,N-dimethylaminopyridine, and cool the solution to 000. Add thereto 5.9 g of p-toluenesulfonyl chloride in portions and stir the so-formed reaction mixture at 000 for 1/2 hour, and then stir it at room temperature for 5 hours. Add 100 mL of EtOAc and suction-filter the mixture. Concentrate the filtrate; add thereto 150 mL of EtOAc, and wash with 5% aqueous KH 2

PO

4 Wash the organic layer with cold aqueous 5% NaHCO 3 then with saturated brine, and then dry it over anhydrous MgSO 4 Filter the mixture, and evaporate the filtrate. Chromatograph the residue on silica gel, 99**eluting with EtOAC-hexane to give 6.4 g of the title compound,

PMR

(CDC1 3 8 7.67(m,5), 7.30(m,6) 6.70(t,2), 4.74(d,1), 4.53(d, 1), 4.13(m,1), 3.97(m,1), 2.43(s,6), 1.95(m,2), 1.77(m,1). Mass spectrum (FAB): 622 EXAMPLE 14 Difluorolhenyl)-2:rr4-methyllhenyl)-su Ifonvloxylmethvfl-5-[ 1 H- (1 .2.4-triazolyfll-l .4-Dentenediol-1 (4-methylbenzene~sulfonate Follow the procedure of Example 13 except substitute an equivalent amount of the product of Example 12 in place of the product of [aPD Example 11 to obtain the title compound, 14.20 MeOH).

EXAMPLE (-)-5R-cis)-5-(2.-4-Difluororohenvl-5-f(1 H-i .2,4-tr azol- 1 -yl' m ethylI-tetrphvd ro.

furanmethanol.4-tolueesul~honate Dissolve 6.3g of the product of Example 13 in 150 mL of toluene and heat the so-formed solution to 10000. Add 2.4g of 60% NaH dispersion in oil portionwise, and then heat the so-formed reaction mixture at reflux until cyclization is complete (approx. 3-4 hours). Cool the mixture and decant the solution from excess NaH. Wash the solution with cold 5% aqueous

KH

2

PO

4 Evaporate the organic layer to form a residue and chromatograph the residue 99 10 on silica gel, eluting with acetone-hexane to obtain 1.6g of the title compound as the less polar of the two products, -3.

0 c1 HI) :9.:PMR (ODC1 3 8 8.09 7.88 7.31 6.81 4.52(ABq,2), 9. 3.99(m,l), 3.85(m,1), 3.70(m,1), 3.59(m,1), 2.49(m,2), 2.47(s,3), 1.90(m,1).

Mass spectrum (FAB): 450 [M+HJ+.

9 EXAMPLE 16 Dilur ~jny)5-H- .2.4-triao- 1 -yl) rnethyll-tetra hyd ro- 3.fu r neth anol4-to Iuenes u I)h on ate Follow the procedure of Example 15, except substitute an equivalent amount of the product of Example 14 in place of the product of Example 13 to give the title compound, 40.30 CHC13), mp 96- 9800.

EXAMPLE 17 (-)-r(2R)-cisl-.44.4[4r[5-(24-difluorrhenyl)-tetrahydro5( 1 H- 1 .2.4-tria2zol-l1-vlmethvl)furan.:3-vIlmetho]dheynvI 1l-cDierazinytlphen-l-2,4dihydro-3H-1 .2.4-triazol-3-one.

The title compound is prepared starting with the tosylate of Example 15 and 4 4 4 -n itrophenyl)- 1 piperazinyl ]phenol (Example 3a of USP 4,791,111) and using the synthetic scheme outlined in Scheme V and J.

Heeres, et al., JMd_ 1984, Vol 27, p894-900 at 898 and 900.

EXAMPLE 18 e n ___-Trh 1 .2.4-Trip zol..1 -vlm ethfl)-3-Fu ra Ly!L1eoxylPhetn ull 1 PinnerazP~ illPh enyLl24- Dihydro-2-r1 (S)-Methyl.2LR)..Hydroxvgro pyl] 3H 1.

2 4 -Tripzol-3..ne.

a. 2-0-S3EM Ether of (2R.3R2.-Bunediol *10 To a stirred solution of 4.95g of (2R, 3 R)-2,3-butanediol, (55 mmoles) and 9.3g of SEM-CI (55.7 mmoles) in 55 ml of anhydrous DMF at 000 were added in four portions 2.34g of 60% oil-dispersed NaH (58.5 mmoles) over 10 min. The resulting mixture was stirred at 000 for 4 hours and at ambient temperature **Ireovernight. The turbid reaction mixture was poured onto 0.5L of 5% KH 2 P0 4 solution and extracted with 2 x 300 ml of ether; the combined ethereal solution was washed once with distilled water, saturated brine, dried over MgSO 4 and evaporated to give a colorless liquid. Flash chromatography over 350g silica gel with 1 L of 7% ETOAC/Hexane, 2L of 10% ETOAC/Hexane and 1 L of @065 ETOAC/Hexane gave 1 .74g of the title compound (yield 14.4%) MS:(M+H)+=221.

A mixture of 0.7g of the 2-0-SEM ether of Example 18(a), (3.18 mmoles) and 0.97g of 4 -bromobenzenesulfonyl chloride (3.82 mmoles) in 5mI of anhydrous pyridine was stirred under

N

2 atmosphere at ambient temperature for 6 hours. The reddish slurry reaction mixture was diluted with 50mI of icecold water, extracted with 2 x 25m1 of ether. The combined ethereal solution was washed with 2 x 25ml of CUS0 4 solution, distilled water, saturated brine, dried over MgSO 4 and evaporated to give a reddish oily residue. Flash chromatography over 50g silica gel with 1L of 10% ETOAC/Hexane gave 1.02g of the brosylate as a colorless liquid (yield 72.9%) 23 [I]D -3.69 (CHCI 3 c=1) c. Alkylation Reaction A mixture of 0.98g of the brosylate of Example 18(b) (2.23 mmoles), 0.69g of the 3H-1,2,4-triazol-3-one of Example 17 (1.12 mmoles) and 0.37g of cesium carbonate (1.12 mmoles) in 20 ml of anhydrous DMF was stirred at 800C under N 2 overnight (-20 hours). The reaction mixture was diluted with 100ml of 10 ice-cold water, extracted with 2 x 50 ml of ethyl acetate. The combined organic solution was washed once with distilled water, saturated brine, dried over MgSO 4 and evaporated to give a brown solid residue. Flash chromatography of the residue over 125g silica gel with 1.2L of 80% ETOAC/Hexane gave 0.327g of the product as a tan solid (yield 35.7%) MS=(M+H)+=81.7.

d. Acidic Hydrolysis of 18(c) to the title oroduct A mixture of 0.32g of the SEM-ether of Example 18(c) and 6ml of 6N HCI solution in 6ml of methanol was stirred at ambient temperature for 4 hours and was evaporated under reduced pressure. The residue was diluted with 5ml of ice water, carefully basified with 10% Na 2

CO

3 solution until pH=8-9 was obtained. Extraction of the so-formed reaction mixture with 2 x25ml of CH 2 Cl2 followed by washing with saturated brine, drying over MgSO 4 and evaporation gave a tan solid. Filtration of the tan solid through a 50g silica gel column and elution with 0.75L of 4% MeOH/CH 2

CI

2 gave 0.26g of title product as a tan -23.65' (CHCI3 c=1) solid, yield 96.6%. MS=(M+H)+=687; D 2365 (CHCI 3 c=1) Example 19 (-)-1(2R)-cil-4-r4-r4-r4-1r(5-(2.4-Difluoro n )T rh ro.5. H- 1.2.4- Triazol-1 -lmethyl)-3-FuranvlMethoxvyPhenvl-1 -PiperpzinvllPhenvl]-2.4- Dihydro-2-11 (R)-Methyl-2(R)-Hydroxvyrovyln-3H. .4-Triazol-3-one a. Mitsunobu Reaction To a stirred solution of 0.72g of the 2-O-SEM ether of Example 18(a) (3.27 mmoles), 2.1g of triphenyl phosphine 8 .06g) and 1.2g of p-nitrobenzoic acid (7.17 mmoles) in 30ml of dry benzene at 0°C were added, dropwise, 1.25ml (8.06 mmoles) of diethyl azodicarboxylate The so-formed 10 clear yellow solution became turbid and the mixture was stirred at ambient temperature for 2 hours, and mixture loaded on a 100g silica gel column.

Elution of the column with 15% ETOAC/Hexane gave 1.5g of the 3-3nitrobenzoate having the S absolute configuration (95% yield) MS: 219 150), 252 117).

b. Basic Hydrolysis of the D-Nitrobenzoate A solution of 1.12g of of the p-nitrobenzoate of Example 19(a) (3 mmoles) and 3.5ml of 1N NaOH solution in 20ml of methanol was stirred at ambient temperature for 3 hours. Solvents were evaporated and the residue was diluted with 10ml of distilled water, and extracted with 2 x 20ml of ether. The combined ethereal solution was washed once with saturated brine, dried over MgSO 4 and evaporated to give 0.67g of the corresponding alcohol as a colorless liquid which was used directly for the next reaction without further purification.

c. Brosvlation, Akvlation and Acidic Hydrolysis Following the procedures of Example 18(c) and the title compound was prepared in 32% overall yield in 3 steps from the products of Example 19(b) [23 23 (CHC3 and of Example 17. MS:

D

-23.65 0

(CHCI

3 c=1) Triazol- 1 -vim ethyl).3-Fura nyl1meth oxvIPh envl1-1-Pi erazinvl1phenlr. 4.

Dihydro-24(S)-1 -MethvI--Hvdroxvlropvll.3H-.1 2 .4-Tripzol-3-one.

a. Formation-of TBDPS Ether To a solution of 0.9g or (R)-(-)-1,3-butanediol (10 mmoles), 1.5g of imidazole (22 mmoles) in 10 ml of anhydrous DMF at 000 were added 3m1 of tbutylchlorodiphenylsilane (OTBDPSN) (11 mmoles) over 3 minutes. The reaction mixture was stirred at 000 for 4 hours, diluted with 50mI of ice-cold water and extracted with 2 x 30ml of ether. The aqueous phase was back extracted with A 50m1 of ether and the combined ethereal solution was washed once with 15 saturated brine, dried over MgSO 4 and evaporated to give a colorless residue.

Flash chromatography over 150g silica gel with 1.5L of 5% EtOAC/Hexane and 1iL of 10% EtOAC/Hexane gave 2.87g of the TBDPS ether (87.5%) 2 3 +0.64' (CHOI3 c=1) MS: 329; b. .BMsxlatIojn To a solution of 0.984g of TBDPS ether of Example 20(a) (3 mmoles) in 7ml of anhydrous pyridine were added 0.845g of 4 -bromobenzenesulfonyl chloride (3.3 mmoles). The reaction was run and worked-up and purified in accordance with the procedure of Example 18(b) and 1.02g of the brosylate was 2 +2.450

(CHOI

3 C=1) obtained in 61.1% yield; MS: [M+23]+s 569/571;

D

Alkylaion The brosylate of Example 20(b), 0-95g (1-74 mmoles) was reacted with the compound of Example 17 according to the procedure of Example 18(c) to provide 0.49g of corresponding alkylated product, yield 60.3% MS: 925 2 3 -3 7 H 3 d. Aci~dicHyrlyi The compound of Example 20(c), 0-32g, (0.35 mmoels) was hydrolyzed *10 by 6N HCI solution in accordance with the procedure of Example 18(d) to give V 0.22g of the title compound (yield MS: M+ 686; 709; ]23 3 Da (0H01 3 C=1) .*Alternatively a solution of 0. 19g of the compound of Example 20(c) and of tetrabutylamrnonium fluoride (0.23 mmoles) in 5ml of THF was stirred at ambient temperature for 24 hours. The brown solution was concentrated to a syrup. Flash chromatography of the syrup over 50g silica gel with 0.51- each of 2% and 4% MeOH/CH 2

CI

2 gave 0.l11g of the title compound (yield 88.7%).

Exa i~he *j 2R)-cis1.4-[4-f4[r4[[s.-(P A-rDifuropenI)erayo( 1H- 1.2.4- Triazo- IYmty)3FrnlMetho]hnl PprznlPhenjyU-..4- The procedures of Example 20 were followed except an equivalent amount of ,3-butanediol was substituted for the corresponding

R

enantiomer. An overall 31.8% yield of the title compound was obtained in four steps; 687.

u n i s b r 4 r 4 f 5 2 4 i f l h5 H 2 4 l 1- i th l :uranl]M -et x v Phen y~ i e a n l en 2.

4 Dhvdo-~r1(S~ M 1-v2(S -Hvdrxyrp 3H 1 rp -3-one.

a. BeQnzylptin To a solution Of 10g of (2R, 3 R)-()-2,3-butanediol (111 mmoles) in of anhydrous

CH

2 01 2 and 80ml of Cyclohexane at 000 were added imi of *.furmehnslfcai (TfOH), followed by dropwise addition of 21m1 o benzyl trichloroacetimidate (11 mls.Terutiglrywa stirred at ambient temperature overnight, diluted with 125ml of hexane and filtered. The combined filtrate was concentrated to a yellow syrup. Flash chromatography of the yellow syrup over 250g silica gel with 1 .51- of 7% ETOAC/Hexane, 2L of 15 15% ETOAC/Hexane and 2L of 25% ETOAC/Hexane, 1 .51- of MeOHICH 2

CI

2 gave 1 l.

8 8g of the 2 -monobenzyl ether of the starting material (74.5% yield) and 2.03g of unreacted starting material MS: 181.

b. Mitsunobu Reaction The 2-monobenzyl ether of Example 22(a), 5.4g, was converted into 6.6g of the 3- benzoate ester (yield 66.9%) by Mitsunobu reaction in accordance with the procedure of Example 19(a); MS: 330.

c. Alkaline Hvdrolysi The 5.3g of the product of Example 22(b) was subjected to alkaline hydrolysis according to the procedure of Example 19(b) to give 2 .33g of the 2monobenzyl ether of 2 R,3S)-2,3-butanediol (yield 80.3%) 181; 10 [a]2 2 3.75 0 (CHCl3 c=1) 10 .d d. Formation of the SEM Ether To a stirred solution of 3.14g of the product of Example 22(c) (17.44 Smmoles) and 3.8ml of di-isopropylethylamine 2 .82g, 21.8 mmoles) in 30ml of anhydrous

CH

2

CI

2 at ambient temperature were added 3.8ml of SEM-CI (3.64g 21.8 mmoles) in one portion. Fuming formed and the resulting yellow solution was stirred for 20 hours. The orange-colored reaction mixture was evaporated under reduced pressure and the solid residues were partitioned between ether and water. The ethereal solution was washed once with distilled water, saturated brine, dried over mg 504 and concentrated to give the crude product.

Flash chromatography of the crude product over 2 00g silica gel with 2L of 3% ETOAC/Hexane gave 5.3g of the 3-O-SEM ether of the product of Example 22(c) (98% yield) as a colorless liquid; MS: [M+HJ+ 311.

e. Hydrogenolvsi A mixture of 5.25g of the product of Example 22(d) (16.94 mmoles) and of 10% Pd/C in 150ml of methanol was hydrogenated under atmospheric pressure for 6 hours. Catalysts were filtered and washed with additional methanol. The combined filtrate was concentrated to give a colorless liquid.

Flash chromatography of the liquid over 100g silica gel with 2L of ETOAC/hexane 3.53g of the free alcohol (yield 95%) as a colorless liquid;

MS:

174, 103.

a f. Brosylation The product of Example 22(e) ig was converted into 1.52g of the corresponding brosylate in 76.2%yield in accordance with the procedure of S..23 18(b); -1.53 (CHCI 3 ;c=1) 18(b); a g. Alkylation Reaciton The brosylate of Example 22(f), 1.48g of was reacted with the product of Example 17 to give 0.75g of the 2-alkylated triazol-3-one (yield 54.3%); [a]D -32.690 (CHCI 3 c=1) a h. Acidic Hydrolysis Hydrolysis of 0.7g of the product of Example 22(g) in accorcdance with the procedure of Example 18(d) gave 0.51g of the title compound as a cream- 23colored solid (yield 86.7%); colored solid (yield MD -32.690 (CHCI 3 c=1) 2 R)-cisl-4-f4r44-.rrs(24.Difluoro)henyI.)Tetrhdrps(I H-1 .2.4- Trao-lYmty'--uranvnlMethoxylPheniyl -PineraZinvlPhey1j2 4- Dihydro-P-f 1 (R)-Methyl-2(S)-HydroxyvroDyll3H-1.

2 4 -Triazol-3-one.

a. Mitsuriobu Reaction The product of step e of Example 22 (1.99g, 9.05 mmoles) was reacted a10 with p-nitrobenzoic acid in accordance with the procedure Example 19(a) to give 3.3g of product (yield MS =221.

0 0 b. Alkaline Hydrolyi 15 The product of step of this Example (2.36g, 6.4 mmoles) was hydrolyzed by 7ml of 1N NaOAc to give 1.18g of the 3-0-SEM ether of (2S,3S)- 2,3-butanediol (yield MS: 221 D(X 2=+55.150

(CHCI

3 C=1) aD c. Brslt omto The product of step of this Example (1.15g were converted into the brosylate in accordance with the procedure of Example 18B(b) to give 3.47g of the brosylate (yield 97.7%).

d. A"'ato anjAiicHyrlysis The procedures of Example 18(c) and were followed except the product of Example 23(c) was substituted for that of 18(b) to give the title compound.

Example 24 (2R-cis-4-f4- 4-14f.-5-14if.dluoroDhenyl)-tetrahydro(1 H-1 4-triazol- 1-Ylmethyl)fura-3-ilmetholphenvenvll2-4-dihydr 1-ethyl-2(S-hydroxvpropyql-31H- 1.2 4 -Triazol--One a. The methyl ester of (S)-lactic acid was converted into the corresponding benzyloxymethyl ether in accordance with the procedure of W. C. Still, et al.

Tetrahedron Letters, 21, 1035-1038 (1980).

b. Reduction to the Aldehyde DIBAL-H, 37.7ml of a 1M solution, was added dropwise to a stirred solution of 7.67g of the ester of step of this Example in toluene at -78 0 C (dry ice/acetone bath) under an atmosphere of nitrogen. After 6 min. methanol S 15 (10ml) followed by an aqueous aolution of Rochelles salt were added. After warming to room temperature the moisture was partitioned between ETOAc and water. The organic phase was separated, washed with water, dried (MgSO 4 and concentrated to produce the crude aldehyde which was used in the next step without purification.

b. Grignard Step The THF solution of 80ml of 1 molar solution of the ethyl magnesium bromide Grignard reagent was added dropwise to a stirred THF solution of the crude aldehyde obtained from step of this Example at -780C (dry ice/acetone bath) under an atmosphere of nitrogen. After the addition was complete, the resulting mixture was allowed to warm slowly to room temperature overnight and stirred for a further.period of 48 h. An aqueous solution of Rochelles salt was added and then the resulting mixture was partitioned between acetone and water. The organic phase was separated, washed with water, dried (MgS0 4 and concentrated. The residue was purified by column chromotography on silica gel using ETOAC/Hexane (1:10) as eluant to give non-polar alcohol (2S,3S) 2.31g;31%, as a colorless oil.

(ii) a mixture of both alcohols, 1.23g; 41% and (iii) polar alcohol (2S,3R) 1.23g; 16%, as a colorless oil.

c. Brosvlation of polar alcohol 10 4 -Bromobenzenesulphonyl chloride (1.035g, 4.1 mmoles) was added to a stirred solution of (0.605g, 2.7 mmoles) the polar (2S, 3R) alcohol of step of this Example and 2.20g (5.9 mmoles) of DMAP in CH 2

CI

2 at room temperature under an atmosphere of nitrogen. The resulting mixture was stirred for 12 h.

and then partitioned between ETOAC and water. The organic phase was 15 separated, washed with water, dried and concentrated. The residue was .:purified by column chromatography on silica gel using ETOAC/Hexane (1:10) as eluant to give the desired brosylate as a colorles oil.

d. Alkylation and acidic hydrolysis The procedures of Example 18(c) and were followed except the (2S, 3R) bosylate of step of this Example was substituted for that used in Example 18(c). The acidic hydrolysis produced the title compound as a white solid, mp 170-1720C.

ExmMl 12R-ci). 44 -105 A2difluo ev).erhdo~ Hi.4-razol- 1ylmethflfuran-lyvflmethow1]Dhenyl..-oieaivfpev 4-2 4- ro-2-rYR}- 1 ethyl -2()hdr oWrp1 3 H1 4-tripar-3on The procedures of Example 24 were followed except the non-polar (2S,3S) alcohol from step of Example 24 was converted into the (2S,3S)-3brosylate. Alkylation of the brosylate followed by acidic hydrolysis of the SEM protecting group in accordance with the procedures of Example 24(d) provided a. 10 the title compound.

V Expmiple 26 (2R-is-4j4[4:[44rL5 2 4 -dif uroh)y-5tp o(l 2. 4-triazol- .l-ylmethyl)furn3ylmethowylhenyll-1i rai hnI91-; *15 etnyl.-2(i)-h ro rOnvI13Hl- 2 trazL-3ne The procedures of Example 24 were followed except the methyl ester of lactic ester was substituted for the methyl ester of (S)-lactic, acid in step of Example 24. The (2R, 3S) alcohol was used in steps and to provide the title compound.

Exam 2_27 12FJ-Q444[4-f.r-5-(2, 4 -difluoroh nyl.ttayr5 1 H-12 -tiz l-zylmethylfra..1me- ,Deyl -Dorzivlhevl4dhdp _S)1 ethl~2R).ydrxy~~2YAF-3H-1 24 -triaK2L-3-pne, The procedures of Example 26 were followed except the (2R, 3R) alcohol was used in steps and to provide the title compound.

Example 28 2R-Bis-.-4-4[4-14- 2-5 f2.4- difliuo r h n l-ttr hH-1 2 4-tri z 1-Yvlmethvl)furan-3-vylmthoxv1henvill -nierazin hen vll2-4-dihvdro ethyl-3-hydroxyvroyvl]3H- 1.

2 4 -triazol-3-One.

a. Reduction To methyl 3 R)-hydroxyvalerate (5.289, 40.0 mmoles) dissolved in 100ml of anhydrous THF at 0-5 0 C was added dropwise 60ml of a 1M THF solution of S 10 LiAIH 4 (60 mmoles). The solution was allowed to warm to ambient temperature and to the so-formed mixture was added sequentially, 2.5 mL of water, dropwise, 2.5mL of 15% NaOH and 7.5mL of water. The so-formed reaction mixture was stirred at ambient temperature for 4 h. The inorganic solids were removed by filtration and the filtrate was evaporated to give 4.31g of (3R)-1,3- 15 pentanediol.

b. 1-0-SEM ether formation .:The procedure of Example 18(a) was followed except an equivalent quantity of the product of step of this Example was substituted for the (2R, 3 R)-2,3-butanediol to provde the title compound.

c. Mitsunobu Reaction The procedure of Example 19(a) was followed except that an equivalent quantity of the product of step of this Example was substituted for the 2-SEM ether of 2 R,3R)-2,3-butanediol to give 3.34g of the corresponding pnitrobenzoate.

d. Basic Hydrolysi The procedure of Example 19(b) was followed except that an equivalent quantity of the p-nitrobenzoate ester of step of this Example was used to provide 1.88g of the 1-0-SEM ether of 3 S)-1,3-pentanediol.

e. Brosy/lationi. Alkylation and Acid Hydrolysis The procedures of Example 18 and were followed except that an equivalent quantity of the product of step of this Example was substituted for the corresponding 1-0-SEM ether of (2R, 3R) 2,3-butanediol used in .9 10 Example 19(b) to produce 1.04g of the title compound of this Example -8.42'

(CHCI

3 C=1)

D

Exmp9e2

POO*

akylte he roduct of..s Exampfleor17hin accorace5 1it th- 1.2.dre 4-raof 9999 15 xaple18c).fThenso-frmed product was sujectiyl~e to aci .dihdrosis 1in accrdnc T h he proceduresan(b of Example 8wrefooe to produce thm(0%yed bo syhe tle byc o llo wn g, thD r c d r s o x m l 8 b. T e 3 r s l t a E ape 8().The so-f-fo-rme(2difodut assujctd o cii hydolyi in accordance with thylle roedure ofExampl 18d)tapovdi38ny90 ied hyr H-i-hjv1-q- 0 2.4-taolazol-e a. Preoaration of (PS)-i 0 -butpnediol A solution of (2S)-3-butene- 1,2-diol which was purchased from Eastman Kodak, (3g, O.O34mmoles) in 4OmL of ethanol was hydrogenated in the presence of 300mg of 10% Pd/C overnight. The so-formed reaction mixture was filtered through celite. The so-formed filter cake was washed with ethanol and the combined filtrates were evaporated to provide 2 .08g (68% yield) of the title compound.

1 0 b 1- E th e r fo rm a tio n b o s at n a k at na da c d ic h d o y i S The procedures of Example 18(a) were followed except that an equivalent amount of the product of step of this Example was substituted for a6 00 00 the (2R, 3R) 2,3-butanediol of Example 18 to provide the title compound 0000 a 23 0506 'D -24.30 (CHCI, 3 c=1) 0 0 Examjple_1 hydoH-i2)-.2.4-triazotrizo-3-n- The procedures of Example 30 were followed except that an equivalent quantity of (2R)-3-butene-i ,2-diol (available from Eastmand Kodak) was substituted for (2S)-3-butene-i ,2-diol in step of Example 30. The procedures of Example 30(b) were there after followed to produce the title [CX] 2 -294 CC3;C1 compound D 294(CC 3 ;c1 Example 32 2 R-cis)-4-f4-4-[4rr-5-(2,4-difluoroDhenvl)tetrahydro-5- H-1,2,4.triazol- -vlmethvl)furan-3-vlmethoxvlDhenvl-1 -DiperazinvllDhenvl-2,4-dihydro- 2-r(S)-1-ethvl-2(S)-hydroxvDrovlI-3H-1 2 .4-triazol-3- ne a. (S)-2-(benzvloxv) DroDionaldehyde by selective reduction of benzyl) lactic acid pyrrolidine amide: To a solution of the S-(O-benzyl)lactic acid pyrrolidene amide prepared in accordance with the procedure of Tetrahedron, 19, vol. 45, pages 57-67 (5g, 0.0214 mol.) dissolved in 20 ml of toluene cooled to in a ice methanol bath was added slowly with stirring 4.25 ml or RED-AL (3.4M solution of sodium bis(2-methoxyethoxy) aluminum hydride) in toluene available from Aldrich Chemical Catalogue #19, 619-3). The solution was stirred fro 5 hrs., quenched with 2.5 ml of acetone and thereafter with 35 ml 15 of 2NHCI. The so-formed mixture was extracted with EtoAc. The organic extracts were washed with water, NaHCO 3 and brine, dired over Na 2

SO

4 and evaporated to give the titled product.

b. (S)-2-(Benzvloxy)-N-(Formvlamino) proDanimine. The propionaldehyde of step (1g, 16.09 mml) was added dropwise to a solution of formyl hydrazine (0.73g, 12.18 mmol) dissolved in 5 ml of methanol. The soformed reaction mixture was stirred overnight. The solvent was removed by evaporation and the so-formed residue was stirred with ethyl ether. The undissolved excess formyl hydrazine was removed by filtration and the ether was removed to provide a residue which was chromatographed on silica gel(/) using 20% EtoAc: hexane to give 805 mg of the title product as a light yellow waxy solid having strong UV activity; ms [M 207.

1i" c. 2-r3-(2s. 3

S)-

2 (Benzyloxy)Den mi aid hydrazide Ethylmagnesium bromide (1.3 ml, 3.9 mmol, 3.0 molar in ethyl ether) was added to a stirred solution of 200 mg, 0.97 mmol of the propanimine of step (b) in 10 ml of ethyl ether at 00C. The so-formed reaction mixture was stirred overnight at room temperature and quenched with water. The organic layer was separated and the solvent removed to provide a residue which was chromatographed on silica gel using 30 to 50% of EtoAc:hexane to provide 113 mg; (50 yield) of the title compound as an oil. The ratio of S,S isomer: 10 S,R isomer in the product was 94:6. When the reaction was repeated in the presence of 1.2 equivalent of bis(trimethylsilyl) acetamide the S,S:S,R ratio improved to 99:1 MS: [M 237 *d.

d. Cvc:izati0n Reactin A solution of 156.3 mg, 0.66 mmol of the product of step and 400 mg 0.60 mmol of 17F of Scheme V and 1 mole of DBU (1,8-diaza bicyclo 9 [5.4.0]undec-7-ere) in volume was stirred at 80°C for six hours; the temperature was raised to 1000 to 11000 and stirring was continued at this temperature overnight. The reaction mixture was allowed to cool to room temperature and the stirring was continued over the weekend. The solvent was removed by evaporation and the crude product was purified on preparative TLC EtoAc) hexane, v:v) to provide 200 mg of the benzyl ether of the title product of this example as a foamy solid; MS:[M 792 This cyclization reaction is the invention of Mergelsberg, Gala et. l which is disclosed in commonly-owned U.s. Patent No. 5,625,064.

e. HIvdrogenolysis To the solution of the benzyl ether (190 mgs 0.24 mmol) of step d dissolved in 10 ml of methanol was added 40 mg of Pd black on carbon and 4 ml of formic acid. The reaction flask was sealed with a ballon and heated at for four hours. The catalyst was removed by filtration through a celite cake and the filtrate was poured into cold water. The pH of the so-formed solution was adjusted to a value of 4 to 5 with amonia. The so-formed mixture was extracted with EtoAc. The organic layer was separted and dried over Na 2

SO

4 10 The solvent was removed to provide a crude product which was purified on preparative TLC methanol: CH 2

CL

2 v:v) to give 95 mg of the title compound of this example. (57% yield) as a tan solid. MS H+ 701.

S[a] -28.4 1.0, CHCl 3 15 ExamDie 3 (-)-f2R)-cis-4-f4-r4-14-ff5-(2.4-Diflurophenvl)-Tetrahydro-5-(1H-1.2.4- T riazol-1- vlmethvl-3-Fu ranvM ethoxyPhenl. 1-PierazinvllPhenvll-24- :Dihydro-2-41 (S)-Ethyl-2(S)-HydroxvoroD.3H1.24.Triazol3one. Ester with Glycine (as Hydrochloride).

A. To a solution of N-Cbz-glydine (315 mg), N,Ndimethylaminopyridine (DMAP, 200 mg), and compound of Example 24 (900 mg) in CH 2

CI

2 (50 mL) at Ooc, add dicyclohexylcarbodiimide (DCCD, 290 mg).

Stir the solution at 00C for 30 min., then at room temperature for 1 hr. Add additional N-Cbz-glycine (700 mg) and then increments of DCCD at 20 min.

intervals until the reaction is complete by TLC. Pour the reaction mixture into aqueous

KH

2

PO

4 and extract with EtOAc. Wash the EtOAc extracts three times with 5% aqueous

KH

2

PO

4 then with brine, and dry the extracts over anhydrous MgSO 4 Filter, evaporate the filtrate, and chromatograph the residue to obtain the N-Cbz-glycinyl ester (1.3 [Mass spec. found: (FAB) 892 B. Stir a solution of the N-Cbz-glycinyl ester of step A above in 100 mL MeOH-96% HCOOH (10:1) in sealed flask with a safety valve. Add 30 mg increments of palladium b lack at 30 min intgervals until the reaction is complete by TLC (6-14 Suction-filter the mixture, add 12 N HCI (0.5 mL) to the filtrate and evaporate the so-formed mixture to dryness. Add water (100mL) and activated carbon (0.8 g) to the residue, suction-filter on a 0.45g nylon 10 membrane. Lyophilize the filtrate to provide 356 mg of the title compound.

[Mass spec. found: (FAB) 795 Example 34 S'Follow the procedure of Example 33 except substitute an equivalent 15 amount of any other N-carbonbenzoxy or N-tert-butoxycarbonyl protected natural amino acid to obtain the corresponding natural a -amino acid ester hydrochloride of the compound of Example 32.

Example (-)-r(2R)-cis1-4-14-f4-4-r5-2.

4 Difluronhenyl)-Tetrahydro5-( 1H-1.2.4- Triazol-1 -vlmethvl)-3-FuranvylMethoxyvPhenyll-1 -PiDerazinvl]Phenvl]-2.4- Dihydro-2-l (S)-Ethyl-2(S)-HydroxyvroDvll.3H-.12.4-Triazol-3-one. Ester with 2 4 -Diaminobutanoic acid (as Di- Hydrochloride salt) Follow the procedure of Example 33 except substitute an equivalent quantity of N,N'-dicarbobenzoxy 2,4-diaminobutanoic acid for N-Cbz-glycine to obtain the title compound.

Example 36 2 R-cis1- -4-r4-r[4rr-(24-DiftjroheyL-_Tetrhydro5(l H-1 .2.4- Triazol- l-yLmetihyl)-3-FuranyrjMe)thoxyjPhenii 1-1 -PioerazinvflPhenul, 4 Dihydro-2-[1 (S)-Ethvl-2(S)-Hvdroxvlropyll-3H 1 .2.4-Triazol-3-one.-Ester with L- Alanine (as Hydrochloride salt) Follow the procedure of Example 33 except substitute an equivalent quantity of N-carbobenzoxy-L-a Ian ine for N-Cbz-giycine to obtain the title compound.

10 Examplle 37 The compounds of formula 20F prepared in accordance with the procedures of Examples 1-32. and Schemes I-VI are listed hereinbelow: H 0 R F G..N j- N ,N 9 9 wherein R, is: Me MeR M+ 701 4sMe Me M+ 701 Me

OH

M+ 701 S/Me

)IIOH

me R M+ 687 Me

~M

OH

M+ 701 Me

OH,

Me

/S

M+ 687 M+ 701 RMe

"OH

Me R M+ 687 sMe S-W OH Me/S M 687 R -Me

-OH

M+ 687 R Me

OH'

M+ 687 S -Me

M-OH

M 687 Me

OH'

M 687 R Me lii'...

the above-listed compound 20F wherein R Me R 9 was prepared by substitution of as prepared by ubsituion of an equivalent amount of 2 R,3R)-2,3-butanediol for the 2 -monobenzyl ether of (2R,3S) butanediol used in step d of Example 22.

The product so formed was treated in accordance with the rocedures of steps f, g, and h of Example 22. The above-listed compound 20F wherein

R

)"«OH

R MeR Ri-- Me was prepared by substitution of an equivalent amount of the 2- O-SEM ether of 2R,3R butanediol for the starting material used in step d of Example 22. The product so formed was thereafter teated in accordance with the procedures of steps c, f, g, and h of Example 22.

In the claims which follow and in the preceding description of the invention, except where the context requires otherwise due to express language or necessary implication, the words "comprises" and "comprising" are used in the sense of "includes" and "including", i.e. the features specified may be associated with further features in various embodiments of the invention.

Claims (21)

1. A compound represented by the formula I r [I] S Sr a *555 wherein X is independently both F or both Cl or one X is independently F and the other is independently Cl; R 1 is a straight or branched chain (C4-05) alkyl group substituted by one or two groups convertible in vivo into one or two hydroxy moieties; or a pharmaceutically acceptable salt thereof.

2. A compound of claim 1 wherein R 1 is a straight or branched chain (C4-05) alkyl group substituted by one or two amino acid ester groups ,each convertible in vivo into a hydroxy moiety.

3. A compound represented by the formula II /R2 'N I N [II] *5. 55 S. S Sr 1 SS* wherein X is independently both F or both Cl or one X is independently F and the other is independently Cl; wherein R 2 is a (C4-C5) alkyl substituted by an amino acid ester group convertible in vivo into a hydroxy moiety; or a pharmaceutically acceptable salt thereof.

4. A compound of claim 3 wherein R 2 is a C alkyl group substituted by an amino acid ester group convertible in vivo into a hydroxy moiety and each X is F. A compound of claim 1 wherein RI is a hydroxy-substituted 04- or 0,5-alkyl group selected from: -CH(C2H)H(R)CH3 -CH( 2 H 5 )CH 2 CH 2 R4, -(CH 2 )f CH(R 4 )0 2 H 5 -CH(CH3)CH(R 4 )CH 3 -C(C2,5)C2R4 and -CH(H 3 )CH 2 CH 2 R 4 wherein R 4 is amino acid ester group convertible in vivo into OH and the carbons with the asterisk(*) have the R or S absolute configuration or a pharmaceutically acceptable salt thereof. a a a a a a *a.a S a*a.

6. A compound represented by formula Ill [fill] wherein R 5 is Me R Me/ RMe MeR S Me Me S 4 Me'M /7Rj Me R11 Me Me F~Me MeS Me S Me R or wherein R 11 is an amino acid ester group convertible in vivo into a hydroxy group, or a pharmaceutically acceptible salt thereof

7. A compound represented by the formula IV c H 2 N Nc [IV] wherein R 9 H(C2Hs)CH(R6)CH3 CH(CH 3 )CH(R 6 )CH 3 wherein R 6 is an amino acid ester group convertible in vivo into OH and the carbons with the asterisk has the R or S absolute configuration or 10 a pharmaceutically acceptable salt thereof.

8. A compound of claim 7 wherein R 6 is a dipeptide ester group.

9. A compound of claim 8 wherein the dipeptide ester group is the ester of glycylglycine, -OCOCH2NHCOCH 2 NH 2 S 15

10. A compound of claim 7 wherein the amino acid ester group is the ester of glycine, i.e. -OCOCH 2 NH 2

11. A compound of claim 7 wherein the amino acid ester group is the ester of 2 4 -diaminobutanoic acid, -OCOCH(NH2)(CH2CH 2 )NH 2

12. The compound of claim 7 wherein the amino acid ester group is an ester of leucine, i.e. -OCOCH(NH2)CH2CH(CH 3 2

13. The compound of claim 7 wherein the amino acid ester group is an ester of isoleucine, -OCOCH(NH2)CH(CH 3 )CH 2 CH3.

14. The compound of claim 7 wherein the amino acid ester group is an ester of valine, -OCOCH(NH2)CH(CH 3 2 The compound of claim 7 wherein R9 CH(C2Hs5)&(R 6 )CH 3

16. The compound of claim 7 or 15 wherein S -CH3 R 9 HC H 3 C

17. The compound of claim 7 wherein the amino acid ester group is O 100 O-C-CH2-N 15

18. A pharmaceutical composition for treating or preventing fungal infection comprising an antifungally effective amount of a compound of any preceding claim together with a pharmaceutically acceptable carrier or diluent therefor.

19. A method of treating and/or preventing fungal infections in a mammal afflicted with same which comprises administering an antifungally effective amount of a compound of any one of claims 1 to 17 sufficient for such The pharmaceutical composition of claim 18 in a form suitable for oral or parenteral administration.

21. A method according to claim 19 wherein the antifungally effective amount of the compound is administered orally or parenterally.

22. A method according to claim 19 which comprises administering a composition according to claim 18 or 1 0 claim .23. Use of a compound of any one of claims 1 to 17 in the manufacture of a medicament for treating and/or preventing fungal infections. a

24. Use of a compound of any one of claims 1 to 17 for the treatment or prevention of fungal infections. ~25. A compound of any one of claims 1 to 17 when used for the treatment and/or prevention of fungal infections.

26. A compound of formula 1 as defined in claim 1 substantially as herein described with reference to any one of Examples 17 to 37. Dated this 10th day of April 2000 SCHERING CORPORATION By their Patent Attorneys GRIFFITH HACK