AU2003200556B2 - Echinocandin derivative and their preparation method - Google Patents

Description

ECHINOCANDIN DERIVATIVES AND THEIR METHOD OF PREPARATION The present invention concerns a novel echinocandin derivatives and their method of preparation.

Accordingly, the invention relates to the compounds of formula (IV): (Iv) wherein R represents the radical

O

8H^17 or the radical

O

J J-OCsH,

R

3 represents a methyl radical; R 4 represents a hydroxyl radical; T represents a hydrogen atom; Y represents a hydrogen atom; W represents a hydrogen atom; Z represents a methyl radical, as well as the addition salts with the acids of the products of formula (IV).

13/10/05,ckl3194oct 3.spcl HO ON

H

3 C HHO NO 'HH O0 CH,

OH

Amongst the addition salts with the acids, those formed with mineral acids, such as hydrochloric, hydrobromic, sulphuric or phosphoric acids or the organic acids like formic, acetic, trifluoroacetic, propionic, benzoic, maleic, fumaric, succinic, tartaric, citric, oxalic, glyoxylic, aspartic, alkanesulphonic, such as sulphonic methane or ethane, arylsulphonic acids like the benzene or paratoluenesulphonic acids can be cited.

In the definition of the substituents, -the alkyl, alkenyl or alkynyl radical is preferably a methyl, ethyl, propyl, isopropyl, nbutyl, isobutyl, terbutyl, decyl or dodecyl, vinyl, allyl, ethynyl, propynyl, cyclobutyl, cyclopentyl, or cyclohexyl radical, -the halogen is preferably fluorine or chlorine or bromine, -the aryl radical is preferably the radical phenyl, -the heterocyclic radical is preferably the pyrrolyle, pyrrolidinyl, pyridyl, pyrazinyl, pyrimidyl, piperidinyl, piperazinyl, quinuclidinyl, oxazoyl, isoxazoyl, morpholinyl, indolyl, imidozoyl, benzimidazoyl, tnazoyle, thiazolyl, azetidinyl, aziridinyl radical, as a salt of the S03H radical, sodium, potassium salts or even the salts of amines can in particular be cited.

Amongst the preferred compounds of the invention: -the compounds of formula (IV) in which T represents a hydrogen atom, -the compounds of formula (IV) in which Y represents a hydrogen atom, -the compounds of formula (IV) in which W represents a hydrogen atom, -the compounds of formula (IV) in which Z represents a methyl radical, -the compounds of formula (IV) in which R 3 represents a methyl radical, 12/02/03,sw 3194spa,2 -the compounds of formula (IV) in which R 4 represents a hydroxyl radical can be especially cited.

Amongst the preferred compounds of the invention, the product of example 14 can be cited.

The compounds of formula (IV) may present significant anti-fungal properties.

They may be active notably on Candida albicans and other Candida like Candida glabrata, krusei, tropicalis, pseudotropicalis, parapsilosis and Aspergillus fumigatus, Aspergillus flavus, Cryptococcus neoformans.

The compounds of formula (IV) may be used as medicines in man or animal, to fight against notably digestive urinary, vaginal or cutaneous candidoses, cryptococcoses, for example neuromenengeal, pulmonary or cutaneous cryptococcoses, bronchopulmonary and pulmonary aspergilloses and invasive aspergilloses of immunocompromise.

The compounds of the invention may be equally used in the prevention of mycosic ailments in people with congenital or acquired immune compromise.

The compounds of the invention are not limited to a pharmaceutical usage, they may be equally used as fungicides in domains other than pharmaceutical.

The invention thus may include, as anti-fungal compounds, the compounds of formula (IV) as well as their addition salts with the acids.

The invention may also be directed to the compounds of formula as medicines.

The invention may very particularly include pharmaceutical compositions containing at least one compound of formula (IV) or one of its addition salts with pharmaceutically acceptable acids as active ingredient.

These compounds could be administered by oral, rectal, parenteral route or by local route by topical application on the skin and the mucous membranes, but the preferred route is the oral route.

They could be solid or liquid and be presented in pharmaceutical forms currently used in human medicine, like for example, simple or sugar coated tablets, capsules, granules, suppositories, injectable preparations, ointments, creams, gels; they are prepared following usual methods. The active ingredient(s) could be incorporated into excipients usually used in these pharmaceutical compositions, like talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous or non aqueous mediums, fatty bodies of animal or vegetable origin, paraffin derivatives, glycol, various diluting, dissolving or emulsifying agents, preservatives.

3 12/02/03,sw13194spa.3 These compositions could equally be presented in the form of a powder intended to be dissolved extemporarily in an appropriate medium, for example apyrogenic sterile water.

The administered dose will vary according to the ailment treated, the subject concerned, the route of administration and the considered product. It could, for example, consist of between 50 and 300mg per day by oral route, in adults for the product of example 14.

The invention is also directed to a preparation process characterised in that a formula (III) compound is submitted HO OH R4 R3 4 NH2 T 0 HN OH (II) HO NH O

Z

W

O

OH

OH

HO Y in which the different substituents retain their prior meaning with the action of an agent capable of replacing NH 2 with NHR, R retaining its prior meaning to obtain the formula (IV) compound.

The present invention is further directed to a compound of the following formula: 12/2//03sw 131 9 4spa,4 N" -OH 1OH

HO

These following examples assist in illustrating the invention without at the same time limiting it.

PREPARATION 1: 1- [N2-(12-m ethyl-1I-oxotetradecyl)-4-oxo-L-ornith in el 4- hydroxyphenyl)-L-threoninej -5-L-serine echinocandin B.

I g of 1- [(4R,5R)-4,5-dihydroxy-N2-( 12-methyl-i -oxotetradecyl)-L-ornithine]4- [4echinocandin B is introduced under magnetic stirring and under nitrogen atmosphere into 25ml of acetonitrile. 455 jgl of trimethylsilyl iodide is added. It is heated at 55 0 C for 40 minutes. It is hydrolysed with a solution of sodium thiosulphate at After 10 minutes of stirring, it is dried under reduced pressure and purified by chromatography on silica. 62% of sought product is obtained.

CCM: rfO .25 (eluent: CH 2 Cl 2 -MeOH-H 2 0 86-13-1).

2/02/03.sw 131 94spa.5 EXAMPLE 1: Trifluoroacetate of 1-(4-amlno-N2-(1 2-methylIoxotetradecyl)-L-orllthine 4-{4-(4-hydroxyphelyl)-L-threonflnl-S-Lserine ochinocandin B (isomer B).

of the product of preparation I is introduced into 2.5m1 of methanol in the presence of 4A activated siliporite. 158mg of ammonium acetate at 200C is added. The obtained solution is heated at 500C and of NaBH3CN is added. It is stirred for 3 hours 15 minutes. I ml of distilled water is added and the solution is concentrated dry. 166mg of product is obtained that is purified by HPLC (C 18 by eluting with the compound

CH

3

CN-H

2 0-TFA (50-50-0.02). 17mg of sought product is obtained.

MH+ 975.

EXAMPLE 2 Trifluoroacetate of I -[4.f2-dimethylamlfloethyl-flhifo-N2- (I 2-mnethyl-I -oxotetradecyl)-L-orflithifleI 4-{4-(4-hyd roxyphenyl)-Lechinocandin B (isomers A and B).

of the product of preparation 1, 1s introduced at 20 0 C into a solution containing I ml of methanol, 160p1 of 2-dimethyl-aminoethylamine, 8ml of a solution I M of hydrochloric acid in methanol in the presence of 4A siliporite. 35mg of sodium cyanoborohydrure is int roduced and stirred for hours at 200C. It is filtered, washed in methanol and concentrated dry. 325mg of product are obtained that is purified by HPLC (018 E) (eluent: CH 3 TFA 45-55-0.02 then CH 3 CN-H20- TFA 42-58-0.02). 8.1lgmg of sought isomer A product and 9.4mg of isomer B sought product are obtained.

Mass Spectrometry: MH1 1046 MNa+ 1068 EXAMPLE 3 Trifluoroacetate of I -[4-[(3-aminopropyl)amlino]-N 2 2-.

methyl-I -oxotetradecyl)-L-ornithile] 4-[4-(4-hyd roxy-phe nyl)-Lne echinocandin B (A and B Isomers).

30cm3 of a 1 M solution of hydrochloric acid is added at 0 0 C in methanol into a solution containing 200mg of the product of preparation 1, 2 ml of methanol and 300pl of diamhinopropane. It is stirred for 15 minutes at 000 and 84mg of sodium cyanoborohydrure at 95% is added. It is stirred for 6 IO 1/02103.swI3 I94spadoc,6 hours at ambient temperature and dried under reduced pressure. The obtained residue is made into a paste in acetonitrile, spun and dried under reduced pressure. 312mg of product that is purified by HPLC (C 1 8 (eluent:

CH

3

CN-H

2 0-TFA 45-55-0.02) and 15mg of isomer A and 10mg of isomer B is obtained.

Mass Spectrometry: MH+ 1032.

EXAMPLE 4: Trifluoroacetate of I -[4-[(4.5-dihydro-1 H-imldazol-2yt)hydrazono]-N2-(I2-methyl-I-.oxotetradecyl)-L-orflithi ne] echinocandin

B.

350mg of the product of preparation1, l12ml of methanol and 130mg of of 2-hydazino 2-imidazoline hydrobromide is kept at reflux for 2 hours whilst stirring. After evaporating dry, 510mg of product is obtained that is purified by chromatography on silica by eluting with the compound CH 2 -C1 2 (86-13-1) then by semi-p reparative (C 1 8 HPLC by eluting with the compound

CH

3

CN-H

2 0-TFA (55-45-0.02). 133mg of sought product is thus obtained.

Mass spectrometry: MH+ 1056 MNa+ 1078 EXAMPLE I -[4-[(2-Hydroxyethoxy) imino]-N2-(1 2-methyl-I oxotetradecyl)-L-ornithl ne] 4-[44-4hydroxyphenyl)-L-threoli serine echinocandin B and corresponding E Isomer.

a mixture of 36mg of O-(2-hydroxyethyl) hydroxylamine, 5mI of ethanol, I 2pl of pyridine, 4pl of pure acetic acid and 1 50mg of the product of preparation I is kept at reflux for 4 hours. 205mg of product that is purified by chromatography on silica by eluting with the. methylene chloride-methanolwater (86-13-1) mixture. 2 products of -rf=0.2 and 0.25 -(isomner Z and- isomer.

E) are isolated.

Mass spectrometry,- MH+ 1033 MNa+ 1055 O0/02103,sw 13 194spa.4oc, 7 EXAMPLE 6: I -[4-(hydroxyimlno)-N2-(I 2-meothyl-I -oxotetradecyl)-Lornithine] 4-[4-(4-hydroxyphenyl)-L-threonlnej-5-L-serifle echinocandin B and corresponding Z Isomer.

A mixture containing 200mg of the product of preparation 1, 8ml of ethanol, 36mg of hydroxylamine hydrochloride is left for 1 hour at reflux whilst stirring. It is dried'and purified by chromatography HPLC (C 18 (eluent

CH

3

CN-H

2 0 60-40). 72mg of Z isomer and 60mg of E isomer is obtained.

Mass spectrometry: MH+ =989 MNa+ 1011 EXAMPLE 7 Trifluoroacetate of I -[4-(hydroxyamino)-N2-(I2-methyl-I oxotetradecyl)-L-omithine] 4-[4-(4-hydroxyphenyl)--threonne]-5-L serine echinocandin B (isomer A and Isomer of E+Z oxime mixture obtained in the previous example, 1cm 3 of trifluoroacetic acid 12mg of sodium cyanoborohydrure at 95% mixture is stirred for 3 hours. It is dried under reduced pressure. It is purified by HPLC (018). The products sought are obtained.

Mass spectrometry: MH+ 991 MNa+ 1013 EXAMPLE 8 Chiorohydrate of I-[(S)-N2-(12-mnethyl-1-oxotetradecyl)- 4-[[(3-piperidinyl)oxy] lmino]-L-ornithine] 4-J4-(4-hydroxyphenyl)-Lserine echinocandin B.

Stage A: 146mg of the product of preparation I and 60plof acetic acid is added to a solution containing 45mg of R-3-(aminooxy)-1-piperidine phenylmethyl carboxylate and 2 ml of methanol. It is stirred for 2 hours at ambient temperature. It is concentrated, purified, by chromatography on silica. by eluting with the 98-2 methylene chloride-methanol compound. The sought product is thus obtained.

Mass spectrometry: I O/02/03,sw 131 94spadoc.8 MH 1206 MNa 1228 Stage B: A compound containing 61mg of the product prepared in stage A, of palladium on carbon and 1 ml of acetic acid is placed under hydrogen atmosphere and stirred vigorously for 5 hours. It is filtered and concentrated. 65% of sought product is obtained.

Mass spectrometry: MH 1072 EXAMPLE 9 Trifluoroacetate of 1-[4-[(2-aminoethyl) aminoJ-N2-(12methyl- -oxotetradecyl)-L-ornithine] 4-[4-(4-hydroxyphenyl)-L.

serine echinocandin B (isomer A and isomer B).

To the solution of 300mg of preparation 1 in 6ml of methanol in the presence of 375pl of ethylenediamine is added 63ml of a solution of 1M of hydrochloric acid in methanol. After 15 minutes of agitation, 126mg of sodium cyanoborohydrure (NaBH 3 CN) is added. The reaction medium is stirred for hours. It is filtered and dried, the products purified by.HPLC (C 18 by eluting with the CH 3 CN H 2 o TFA (40-60-0.02) mixture. The sought products are thus obtained.

Mass spectrometry: MH 1018 MNa 1040 EXAMPLE 10: 1-[4-[(2-bromoethoxy) imino)-N2-(12-methyl-1oxotetradecyl)-L-ornithine] 4 4 4 serine echinocandin B and corresponding Z Isomer.

402mg of bromo-2-ethoxyamine bromhydrate is added to a solution containing 710mg of the product of preparation 1 and 28ml of absolute methanol. The mixture is brought to reflux for 55 minutes. It is concentrated under reduced pressure. The obtained product is purified by flash chromatography on silica by eluting with the methylene chloridemethanol compound. The sought products isomer A: Rf=0.54, isomer B: Rf 0.47 are obtained.

10/02/03.sw l3194spa.doc.9 *Mass spectrometry: MH+ 1095 MNa+ 1117 EXAMPLE 11 I± Trifluoroacetate of 1.[4-[(amlnolminomethyl) hydrazono]-N2(1 2-methyl-I -oxotetradecyl)-L-ornlthine] serine echinocandin B.

162mg of aminoguanidine hydrochloride is added to a solution containing 260mg of the product of preparation 1 and I Oml of n-butanol. The reaction medium is brought to reflux for 2 hours 30 minutes. It is concentrated under reduced pressure. The obtained product is purified by semi-preparative H PLC. 225mg of product in a 50/50 mixture of isomers is obtained.

Mass spectrometry: MH+ 1030 MNa+ 1052 EXAMPLE 12: Trifluoroacetate of (dimethylamino)ethoxyimino]-N2-(1 2-mnethyl-I ,oxotetradecyl)-Lornlthine] 4-[4-(4-hydroxyphenyl)-L-threonine]-5-L-serine echinocandi n B and corresponding E isomer.

80.5mg of the product of example 10 are introduced into 32ml of an ethanolic solution of dimethylamine. The reaction medium is brought to reflux for 45 minutes. It is concentrated. The obtained product is purified by HPLC

(C

18

(CH

3

CN-H

2 0 TFA 60-40-0.02). The sought products are thus.

obtained.

Mass spectrometry: MH+ 1060 EXAMPLE 13 Trifluoroacetate of .1-[4-[[2-aminoethoxy)-imino]j-N2- (12-methyl-I -oxotetradecyl)-L-ornithine] 4-[4-(4-hydroxyphenyl)-Lthreonine]-6-L-serine echinocandin B and corresponding Z isomer.

of the product of example -10 is introduced into ammonia. It is.

stirred under pressure for 16 hours whilst allowing it to come to ambient temperature. The reaction medium is again placed in the (45-55)

CH

3 CN-H20 compound to be purified by HLPC (C 18 The sought products are obtained.

IO/0Y103,sw I 94spa.doc, Mass spectrometry: MH+ 1032.

Preparation 2: deoxymulundocandin "nucleus" 2g of deoxymulundocandin are dissolved in 20ml of DMSO. This solution is poured into a suspension containing 120g of FH2264 Utahensis actinoplanes in 870mi of a KH2PO4, K2HPO4 (pH: 6.8) buffer. The reaction medium is stirred for 70 hours at 30 0 C. It is filtered. The mycelium is washed with the phosphate buffer (pH: The washing liquids and the filtrate are joined. The obtained product is chromatographed on a DIAION HP 20, resin and a product is obtained that is used as hereafter.

EXAMPLE 14: Trifluoroacetate of 1-[4-[(2-aminoethyl) amino] (octyloxy)[1.1'-biphenyU-4-yl]carbony-L-ornithine] serine echinocandin B (isomer A) Stage A: 1-[(4R,5R)-4.5-dihydroxy-N2-[[4'-(octyloxy)[1.1'-biphenyl]-4yl]carbonyl]-Lomithine]-4-[4-hydroxyphenyl)-L-threonine]-5-L-serine echinocandin B 1-Preparation of the ester 632g of 2.3.4.5.6 pentafluorophenol is added in 695mg of N, N'dicyclohexylcarbodiimide to Ig of 4'-octyloxy-[1.1'-biphenyl]4-carboxylic acid in 22ml of tetrahydrofurane, stirred for 22 hours at'ambient temperature, filtered, the solvents are eliminated under reduced pressure, the residue is placed into ether, stirred at about 35 0 C, filtered, the solvent is evaporated, it is dried and 1.46g of expected product is obtained, used as it is.

2-Coupling 677mg of deoxymulundocandin <<nucleus>> obtained in preparation 2 is introduced, into 16m1 of DMF. The obtained solution is stirred for 5 minutes and 793g of 4'-(otyloxy)-[1.1'-biphenyl-4-pentafluorophenyl carboxylate obtained above is added.

The reaction compound is stirred and kept under nitrogen atmosphere for 24 hours. It is filtered and concentrated. The residue is placed into ether, triturated, stirred for 25 minutes, spun, washed with ethylic ether, chromatographed on silica by eluting with the (86/13/1) then (80/20/1) 10/02/03,sw I3194spa.doc, II netlene chloride, methanol, water mixture. The sought product is thus obtained. Yield 73%.

Stage B: I -[N2-[[4'-(octyloxy)-[I .1 6biphenyl]..4.yllcarbonyl].4-oxo-L-ornithineF- 4-4(-yrxpey)Ltronnl5Lsdeehncni

B

311lpl of trimethylsilyt iodide is added to a suspension containing 809mg of the product of stage A and I19m1 of acetonitrile. The reaction medium is stirred for minutes at 600C under nitrogen atmosphere. The compound is poured into a sodium thiosuiphate saturated solution. The residue obtained is evaporated and chromatographed on silica, by eluting with the 86113/1 methylene-Chioride methanol water compound. The sought product is obtained. Yield Stage C: I -[4-[(2-aminoethyt) amino-N2-[[4-(octYloxyfl .1 '-biphenylU-4yl]carbonylLomihine-44(4-hydrox~yphen)Lthreonine]5L serine echinocandin B (iso mer A) trifluoroacetate 560p1 of acetic acid are added to a solution containing 900mg of the product of t he preceding stage, 16m1 of methanol and 250pl of diamine ethylene. It is stirred for 15 minutes and 64mg of sodium cyanoborohydrure is added. It is stirred for 18 hours. It is filtered and concentrated. The residue is placed in a minimum of water, triturated, spun and purified by preparative HPLC by eluting with the compound

CI-

3

GNIH

2 OITFNI (55-45-0.2). The sought product is obtained. Yield 26%.

Spectrum RMVN CDC13 9.07 (m wide) I H; 8.48 (dl, J=8) I H ;8.00 (dl, J=8) 2H ;7.96 (dl, J=8.5) 2H; 7.71 (dl, J=8.5) 2H 7.64 (dl, J=8.5) 2H 7.60 (dl, J=9) I1H 7.37 (dl, 1IH 7.02 (dl, J=8.5) 2H 6.97 (dl, J=8.5) 2H 6.65 (dl, J=8.5) 2H 4.90 (in) 1K; 4.77 (in) 1K 4.66 (in) 1H 4.45 (mn) 1H 4.42 (in) 1H 4.39 (mn) 1H; 4.34 (sI) 1IH 4.26 (in) I1H-; 4.22 (mn) I H 4.08 (in) I H 4.01 2H 3.88 (in) 3H 3.70 (in) 2H 3.51 (in) 2H 3.48 (in) I H 3.31 (in) 2H 3.28 (in) IH; 3.16 (in) 2H 2.53 (dd, J=6 et 13.5) 1IH 2.44 (dd, J=7.5 et 13.5) 11-R; 2.27 (in) 1KH; 2.25 (in) I H 2.15 (in) 2H 1.94 (in) I1H; 1.74 (in) 2HW 1.44 (in) 2H; 1.22 to 1.40 (in) 8H; 1.13 J=6) qH J=6.5) 3H 0.88 J=7) 3H.' EXAMPLE 15: 1 -[4-(aminolin omethyl)hydralU-fl 2 U1 4 -4 4 (pentyloxy)-phefl]-i- piperazinyU~pheflcarbofylYLornithne] 4 4( 4 hydroxyphenyl)-Lthreofiel--Lserine-echinocandin

B

Stage A: 5R)-4 .5-dihydroyN-[4[-4(pnyoy hnlpiperazinyl phe nyl]carbofylYLrnithine4(4hydrop henyl) Lthreo ni]echinocandin

B

I O/02/03.ssv I31 94spa.doc. 12 1-Preparation of the ester of pentafluorophenol and 61mg of N, N' dicyclohexyl carbodiimide is added to a mixture of 100mg of [4-[4-[4-(pentyloxy)phenyl]-1piperazinyl]phenyl]carboxylic acid and 3ml of tetrahydrofurane. The reaction compound is stirred at 20"C for 16 hours, filtered, washed with THF and concentrated dry. It is placed in diethylic ether, filtered, washed and concentrated. 71mg of product is obtained.

2-Coupling A suspension.containing 71mg of the ester above, 70mg of deoxymulundocandin <<nucleus>> obtained as in preparation 2, 2.5ml of DMF in the presence of 4A activated siliporite is stirred at 20°C for one night.

It is concentrated, the product obtained is placed in ether and filtered. A product is obtaired that is chromatographed on silica by eluting with the mixture acetonitrile/water/trifluoroacetic acid (60-40-0.02). 30mg of sought product is thus obtained.

Stage B: 1-[N2-[[4-[4-[4-(pentyloxy)phenyl]-1-piperazinyl]-phenyl]-carbonyl]4oxo-L-omithine]-4-[4-(4-hydroxyphenyl)-L-threonine]-5-L-seine echinocandin

B

1-Preparation of the ester a mixture of Ig of the product of stage A, 25 ml of acetonitrile, in the presence of 4A activated siliporite is heated to 55 0 C. 430ml of trimethylsilane iodide is added. It is stirred for 45 minutes then 150pl of an aqueous solution of sodium thiosulphate at 30% is added. It is stirred for 40 minutes at 20*C and concentrated. The dry extract is placed in water, stirred for 1 hour at spun and washed. A product is obtained that is chromatographed-on silica by eluting with the compound methylene chloride-methanol-water (86/13/1).

497mg of sought product are obtained. Yield 42%.

Stage C: 1-[4-[(amirioiminomethyl)hydrazono]-N2-[[4-[4-[4-(pentyloxy)phenyl]- -piperazinyl]phenyl]carbonyl]-L-ornithine]-4-[4-(4-hydroxyphenyl)-L-

B

A suspension containing 400mg of the product of stage B,.4.8ml of n-butanol and 221 mg of aminoguanidine hydrochloride is heated at 1300C for 3 hours.

It is concentrated and 705mg of a product is obtained that is chromatographed on silica by eluting with the methylene chloride methanol 10/02/03,sw 13194spa.doc, 13 compound 85/15, then by semi-preparative HPLC (kromasil C18) with a 40.60.0.02) acetonftrilelwater/trifluoroacetic acid compourid. 64mg of sought product Isthus obtained.

Spectrum RMN CDC1 3 10.75 (s 0.66H 10.45 s) 0.33H 8.39 J=8 0.33K 8.34 m) IH; 8. 10 d, J=7.5 0.66H ;8.08 J=8 0.33H 7.99 d, J=8.5 0.66H 7.74( d, J=8.5 1.33H ;7.71 J=8.5 0.66H ;7.60 J=8.5 0.66K 7.50 m) 1.33H 7.00 m )6H ;6.86 d, J=8.5) 2H ;6.65 J=8 2H 5.08 dt, J=2 et 11.5) 0.66H 4.94 1H 4.88 0.33H 4.75 (din, J=8) 0.33KH;4.67 dd, J=3 et 7.5 0.66H ;4.43 (in 1 H 4.38 (in 1 .66H ;4.33 (irn) 0.66H ;4.26 to 4.20 heavy 2.33H 4.12 d, J=9 0.66K ;4.00 to 3.68 heavy) 7.33H 3.90 t,J=7 2H ;3.62( d,J=1 2 0.33H ;3.43 (swide) 2H ;3.30 to 3.20 (in) I H ;3.20 swide 2H ;2.91 d,J=14 )0.66H ;2.86 (in 0.33H 2.76 (in 0.33H; 2.63 dd, J=1 4 et 12.5 0.66H dt, J=6 et 13 I H 2.44 dd, J=8 and 13 1H ;2.35 (in 0.33H 2.25 m) 1.66K 1.93 (tWde,J= 13 I H; 1.69 (mn 2H; 1.42 to 1.30 (heavy) 4H ;1.15 J=6) 1.98KH;1.10 (J=6 0.99H 0.98 d, J=6.5 3H 0.90 3H.

EXAMPLE 16: 1 -[4-[(2-amlnoethyl)amlnoj412-[4-[4".{pentyioxy)[I 1"terphenyU-4-yI]carbonyl]-L-ornlthlne]-4-[4.(hydroxyphenyl)-L- B (isomer A and Isomer. B).

Operating as previously, from deoxy-mulundocandin <<nucleus>> prepared as indicated in preparation 2 by obtaining 5R)-4.5-dihydroxy-N2-[[4"- (pentyloxy)[1 1"-terphenyl]-4-yI]carbonyl]-L-omithine]-4-[4-(4- B and the corresponding 4-oxo derivative as intermediate product, the sought-product is obtained.

Spectrum RMN QDCI 3 9.00 (wide) 1KH; 8.37 (dl, J=8.5) 1KH; 8.28 I H; 8.10 (dl, J=6) I H 8.02 (dl, J=8) 2K; 7.82 (mn) 4H; 7.73 (di, J=8) 2H; 7.66 (dl, J=8) 2H; 7.38 (dl, J=9) 1KH; 7.32 (dl, J=9) 1K; 7.03 (dl, J=8.5) 2HK; 6.96 (dl, J=8) 2H 6.66 (dl, J=8) 2HK; 5.03 (in) 1K; 4.84 (in) 1KH; 4.67 (mn) I H; 4.45 (mn) 2HK; 4.36 (dd, J=7.5 and 10.5) 1KH; 4.23 (mn) 2H 4.18 (sI) 1K; 4.04 (in) 1K; 4.02 J=6.5) 2HK; 4.00 (in) 1KH; 3.87 (dl, J=9.5) 1K; 3.76 (in) 1K; 3.72 (in) 2K; 3.55 (in) 1K; 3.44 (in) 1K; 3.35 (in) 2H; 3.30 (mn) 1K; 3.19 (in) 2K; 3.12 (in) 1K; 2.53 1K; 2.45 (in) 1K; 2.12 to 2.30 (in) 3H; 1.90 to 2.05 (in) 2K; 1.74 (in) 2K; 1.30 to 1.55 (in) 4K 1.20 J=5.5) 3K 0.96 J=6.5) 3K 0.91 J=7) 3H.

EXAMPLE: Pharmaceutical comp~osition: Tablets have been prepared containing: -The product of example 150mg -Excipient ig (Detail of the excipient: starch, talc, magnesium stearate).

I O/02/03.ssv13 194spa.doc. 14 PHARMACOLOGICAL STUDY A Inhibition of the glucane synthesis of Candida albicans.

The membranes of Candida albicans are purified according to the process described by Tang and al Antimicrob. Agents Chemother 35, 99-103, 1991. 22.5gg of membrane proteins are incubated in a mixture of 2Mm of 14C-UDP glucose (specific activity 0.34mCi./mmol, 50Rg of a-amylase, 1Mm of dithiotreitol (DTT), 1Mm EDTA, 100Mm NaF, 7LM of GTP-y-S, 1M of sucrose and 50Mm of Tris-HCL (pH 7.8) in a volume of 100 The medium is incubated at,25°C for 1 hour and the reaction terminated by addition of TCA to a final concentration of The reaction medium is transferred onto a prehumidified glass fibre filter. The filter is washed, dried and its radioactivity is counted.

Mulundocandin is used as positive control.

The control of the medium is carried out with the same quantity of DMSO The obtained results show that the products of the invention present a good activity in this test, particularly the products of example 14.

B activity on the enzyme Aspergillus fumigatus.

The enzyme is prepared according to the Beaulieu et al. (Antimicrob. Agents Chenother 38, 937-944, 1994) process.

The protocol used is identical to the protocol described above for the enzyme Candida albicans except that dithiotreitol is not used in the reaction medium.

The products present a good activity in this test.

This is a divisional application of Australian Patent Application No. 15659/99, the disclosure of which is, incorporated herein by way of reference.

12/02/03,sw13194spa.

Claims (2)

16- The claims defining the invention are as follows: 1. The compound of formula (IV): QV~ wherein R represents the radical 0 or the radical R 3rcprcscnits a meithiyl radical;, R 4 rcprcscrits a h-ydroxyl radical IT represenits a hydrogeni atomr; Y represenits a hiydrogeni atomi; W represenits a hiydrogeni atomn; Z represenits a mrethyl radical, as wAcll its the addition salts w)(th the IcHis of1C the podcIItS Of formu11la I k/ I OW I I I A 0; 16

17- 2. A process of preparing a compound of formula as defined in any one of claims 1 to 7 which process comprises submitting a compound of formula (III) pIH) in which the different substituents retain their prior meaning to the action of an agent capable of replacing NH 2 by NHR, R retaining its prior meaning to obtain the compound of formula (IV). 3. Preparation process of claim 2 which process is substantially as herein described. DATED this 13th day of October 2005 AVENTIS PHARMA S.A. By their Patent Attorneys: CALLINAN LAWRIE 13/10/05,ck 3194oct13.clnims,1 7