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The present invention relates to compounds with antimycotic activity for human and veteri- nary use and, more particularly, it relates to azole compounds with antimycotic activity useful for the treatment and prophylaxis of infections caused by fungi and yeasts in human and in animals.

Among the antimycotic compounds known in the literature, the so-called azole derivatives among which there are some compounds used in therapy such as Fluconazole (The Merck Index, XI ed., No. 4054, page 645), Itraconazole (The Merck Index, XI ed., No. 5131, page 825) and Ketoconazole (The Merck Index, XI ed., No. 5181, page 835) represent a relevant portion.

As far as we know, however, none of these compounds is endowed with a significant antimy¬ cotic activity against some pathogenic opportunistic fungal strains which induce infections, sometimes fatal, in immunodepressed patients.

Among the azole derivatives known as antimycotic for human or veterinary use, several -com¬ pounds have been described which are characterized by the presence of a tertiary alcohol function of formula

wherein Az is a triazole or imidazole group, X is preferably -chlorine, fluorine or tnfluoro- methyl, R is preferably hydrogen, -chlorine or fluorine, R' and R", the same or different, are hydrogens or alkyls, Y is S, SO, SO2 or O and A is alkyl; which hereinafter will be indicated with the term azole tertiary alcohols.

Among these azole tertiary alcohols we cite, for example, the compounds described in the

European patent applications No. 54974 (Sumitomo Chemical Company Limited), No. 61835 (Imperial Chemical Industries PLC), No. 107392 (Pfizer Limited), No. 140154 (Sumitomo

Chemical Company Limited), No. 178533 (Sumitomo Pharmaceutical Company Limited),

No. 435081 (SS Pharmaceutical Co. Ltd.) and No. 473387 (Sankyo Company Limited). For some of these compounds a remarkable antimycotic activity for topic as well as for sys¬ temic use has been reported, sometimes generically. However, as far as we know, the only compound under study is a compound known as Genaconazole, (2R,3R) α-(2,4-difluoro- phenyl)-α-[l-(medιylsulfonyl)ethyl]-lH-l,2,4-triazol-l-eth anol, described in the European patent application No. 178533.

Very recently, a class of azole tertiary alcohols (I) with antimycotic activity wherein R' and R" are fluorine atoms, Y is a simple bond and A is a diftuoromethyl group has been described [Japanese patent application No. 07/70087 in the name of Taisho Pharma Co. Ltd. - Chemical Abstracts, vol. 123, No. 228190 (1995)]. We have know found that a class of azole derivatives of formula

w erein

R1 is chlorine, fluorine or trifluoromethyl;

R2 is hydrogen, chlorine, fluorine or trifluoromethyl;

Z is CH or N;

R3, R4 and R5, the same or different, are hydrogen or Cj-C4 alkyl, provided that R4 is dif- ferent from R5 when R3 is hydrogen;

X is O or S;

R _« 5 is a C1-C5 polyfluoroalkyl group containing at least two fluorine atoms and optionally other halogen atoms selected among chlorine and bromine; described in the European patent application No. 315946 (Presidenza del ^" Consiglio dei Mini- stri - Ufficio del Ministero per il Coordinamento delle miziative per la Ricerca Scieπtifica e

Tecnologica) as compounds for agriculture use having immunizing activity against fungal pathogenesis and phytogrowth regulating activity towards useful growings, is endowed with a

potent antimycotic activity against a wide range of pathogenic fungi in human and in animals, in particular also against fungal strains resistant to antimycctics used in therapy and against opportunistic pathogenic fungal strains responsible for infections in immunodepressed pa¬ tients, and is active for topical as well as for systemic route. Therefore, object of the present inventions are compounds of formula

wherein R s chlorine, fluorine or trifluoromethyl;

R ₂ is hydrogen, chlorine, fluorine or trifluoromethyl; Z is CH or N;

R3, R4 and R5, the same or different, are hydrogen or C1-C4 alkyl, provided that R4 is dif¬ ferent from R5 when R3 is hydrogen; X is O or S;

Rό is a C2-C5 polyfluoroalkyl group containing at least two fluorine atoms and optionally other halogen atoms selected among chlorine and bromine; and their salts with pharmaceutically acceptable acids; the -compounds l-( 1H- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-dichlorophenyl)-3-methyl-4- ( 1,1,2,2- tetrafluoroethoxy)butane, l-(lH-l,2,4-triazolyl)-2-hydroxy-2-(2,4-dichlorophenyl)-3-me thyl- 4-(l,l,2,2-tetrafluoroethoxy)hexane, l-(lH-l,2,4-tetrazolyl)-2-hydroxy-2-(4-chlorophenyl)- 3,3-dimethyl-4-(l, l,2,2-tetrafluoroethoxy)butane and l-(lH-l,2,4-triazolyl)-2-hydroxy-2-(4- chlorophenyl)-3-methyl-4-(l,l,2,2-tetrafluoroethoxy)butane being excluded. A further object of the present invention are the compounds l-(lH-l,2,4-triazolyl)-2-hydroxy- 2-(2,4-dichlorophenyl)-3-methyl-4-< 1 , 1 ,2,2-tetrafluoroethoxy)butane, l-( 1 H- 1 ,2,4-triazolyl)- 2-hydroxy-2-(2,4-dichlorophenyl)-3-methyl-4-(l , 1 ,2,2-tetrafluoroethoxy)hexane, 1 -( 1H- 1 ,2,4- tetrazolyl)-2-hydroxy-2-(4-chlorophenyl)-3,3-dimethyl-4-( 1 , 1 ,2,2-tetrafluoroethoxy)butane

and l-( 1H- 1 ,2,4-triazolyl>2-hydroxy-2-(4-chlorophenyl)-3-methyla -( 1 , 1 ,2,2-tetrafluoroeth- oxy)butane, specifically exemplified in the European patent application No. 351946, for use as a medicament. For the sake of simplicity, all the compounds object of the present invention comprised the compounds for use as a medicament only will be indicated hereinafter as compounds of for¬ mula π.

The compounds of formula II are endowed with a potent wide-range antimycotic activity, in particular against Candida spp and Cryptococcus neqformans strains, which are Fluconazole and Itraconazole-resistant, and against Candida glabrata, Candida krusei, Aspergilhis spp and Fusarium spp, which are Itraconazole-resistant and, as the previous ones, pathogenic strains responsible for mycotic infections in immunodepressed patients, and are useful for the treatment and the prophylaxis of infections caused by fungi and yeasts in human and in ani¬ mals. The compounds of formula II contain at least a chirai center and then they can be in the form of stereoisomers.

The single stereoisomers of the compounds of formula II as well as their mixtures fall within the present invention. With the term C1-C4 alkyl among the meanings of R3, R4 and R5, we intend methyl, ethyl, n.propyl, isopropyl, n.butyl, isobutyi, sec.butyl and t.butyl groups, methyl and ethyl being pre¬ ferred.

With the term C1-C5 polyfluoroalkyl group containing at least two fluorine atoms we prefer¬ ably intend difluoromethyl, trifluoromethyl, l,l,2-trifluoro-2-chloroethyl groups, 1,1-di- fluoroethyl, 2,2,2-trifluoroethyl, 1,1,2-trifluoroethyl, 1,1,2,3,3,3-hexafluoropropyl, 2,2,3,3- tetrafluoropropyl, 1 , 1 ,2,2-tetrafluoroethyl and their isomers; the 1 , 1 ,2,2-tetrafluoroethyl group being still more preferred.

The salts of the compounds of formula II are salts with pharmaceutically acceptable organic or inorganic acids such as chloridric, bromidric, iodidric, sulfuric, nitric, fosforic, acetic, ox¬ alic, maleic, benzoic, benzenesulfonic, 4-methvlbenzenesulfonic, fumaric, lactic, tartaric, citric and gluconic acid.

Preferred compounds of formula II are compounds wherein R\ is chlorine or fluorine, R is

hydrogen, chlorine or fluorine, R3 is methyl, R4 and R5, the same or different, are hydrogen, methyl or ethyl, Z is N and Rg is a 1, 1,2,2-tetrafluoroethyl group.

Still more preferred compounds of formula II are compounds wherein Ri is chlorine or fluo- rine, R ₂ is hydrogen, chlorine or fluorine, R3 is methyl, R4 and R5, the same or different, are hydrogen, methyl or ethyl, Z is N and R^ is a 1,1,2,2-tetrafluoroethyl group and X is O.

Specific examples of preferred compounds of formula II are the following:

H IH- 1 ,2,4-triazolyl 2-hydroxy-2-(2,4-dichlorophenyl)-3-methyl-4-( 1 , 1 ,2,2-tetrafluoroeth- oxy)butane H IH- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-dichlorophenyl)-3-methyl-4- ( 1 , 1 ,2,2-tetrafluoroethyl- thio)butane

1 -( 1 H- 1 ,2,4-triazolyl>2-hydroxy-2-(2,4-difluorophenyl)-3-methyl- 4-( 1 , 1 ,2,2-tetrafluoroeth- oxy)butane

1 -( IH- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-difluorophenyI)-3-methyl-4- ( 1 , 1 ,2,2-tetrafluoroethyl- thio)butane

1 -( IH- 1 ,2,4-triazolyl)-2-hydroxy-2-(4-chlorophenyl)-3-methyl-4-( 1 , 1 ,2,2-tetrafluoroethoxy)- butane l-(lH-l,2,4-triazolyl)-2-hydroxy-2-(4-dιlorophenyl)-3-methy l-4-(l,l,2,2-tetrafluoroethyl- thio)butane 1-(1H- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-difluorophenyl 3,3-dimethyl-4-( 1 , 1 ,2,2-tetrafluoro- ethoxy)butane

1 -( 1 H- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-difluorophenyl)-3 ,3-dimethyl-4-( 1 , 1 ,2,2-tetrafluoro- ethylthio)butane

1-(1H- 1 ,2,4-triazolyl 2-hydroxy-2-(4-fluorophenyl)-3-methyl-4-( 1 , 1 ,2,2-tetrafluoroethoxy)- butane l-(lH-l,2,4-triazolyl)-2-hydroxy-2-(4-fluorophenyl 3-methyl-4-(l,l,2,2-tetrafluoroethyl- thio)-butane

HlH-l,2,4-triazolyl 2-hydroxy-2-(4-achlorophenyl)-3,3-dimethyl-4-(l,l,2,2-tetraf luoroeth- oxy)butane 1 -( 1H-1 ,2,4-triazolyl)-2-hydroxy-2-(4-chlorophenyl)-3,3-dimethyl-4- ( 1 , 1 ,2,2-tetrafluoroethyl- thio)butane

l-( IH- 1 ,2,4-triazolyl 2-hydroxy-2-(4-fluorophenyl)-3 ,3-dimethyl-4-( 1 , 1 ,2,2-tetrafluoroeth- oxy)butane

1 -( 1 H- 1 ,2,4-triazolyl>2-hydroxy-2-(4-fluorophenyl>3 ,3-dimethyl-4-( 1 , 1 ,2,2-tetrafluoroethyl- thio)butane

H IH- 1 ,2,-4-triazolyl)-2-hydroxy-2-(2,4-didιlorophenyl)-3-methyl- 4-( 1 , 1 ,2,2-tetrafluoroeth- oxy)hexane l-( IH- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-dichlorophenyl)-3-methyl-4- ( 1 , 1 ,2,2-tetrafluoroethyl- thio)hexane l-( IH- 1 ,2,4-triazolyl 2-hydroxy-2-(2,4-difluorophenyl)-3-medιyl-4-( 1 , 1 ,2,2-tetrafluoroeth- oxy)hexane l-(lH-l,2,4-triazolyl)-2-hydroxy-2-(2,4-difluorophenyl)-3-me thyl-4-(l,l,2,2-tetrafluoroethyl- thio)hexane l-( IH- l,2,4-triazolyl)-2-hydroxy-2-(2,4-dichlorophenyI)-3,3-dimeth yl-4-( 1 , 1 ,2,2-tetrafluoro- ethoxy)butane

1 -( 1 H- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-dichlorophenyl)-3,3 -dimethyl-4-( 1 , 1 ,2,2-tetrafluoro- ethylthio)butane

The preparation of the compounds of formula II can be carried out according to synthetic processes described in the already cited European patent application No. 315946. An alternative method for the preparation of the compounds of formula π wherein X is O and

R^ is a trifluoromethyl group consists in treating an alcohol intermediate of formula

wherein Ri , R2, R3, R4, R5 -and Z have the already reported meanings and X is O, with car- bon disulphide and methyliodide under basic conditions and in the subsequent fluorination of the resultant methyldithiocarbonate. The separation of the stereoisomers of formula II can be carried out with conventional tech-

niques such as fractionated crystallization and chromatography.

The preparation of the salts of the compounds of formula II can be carried out according to conventional techniques, for example by mixing in solution equimolar amounts of the -compound of formula II and of the selected acid and by separating the salt by predpitation and filtration or by evaporation of the solvent.

The compounds of formula II and th r salts are antimycotic compounds useful for the treat¬ ment and the prophylaxis of infections caused by fungi and yeasts in human and animals. In fact, the compounds of formula II , object of the present invention, are endowed with an- timycotic activity against yeasts, filamentosous fungi, dermatophyti and dimorphic fungi.

The antimycotic activity has been evaluated in vitro as IC50 and as MIC (Minimum Inhibiting Concentration) against numerous strains such as, for example, Candida albicans, Cryptococ- cus neoformans, Trichophyton mentagrophytes, Aspergillus fiimigatus, Candida parapsilo- sis, Candida lusitaniae, Candida kejyr, Candida tropicalis, Candida krusei, Candida glabrata, Aspergillus niger and Fusarium spp.

It is worth underlining that the compounds of formula LL object of the present invention, re¬ sulted to be active against all the tested Candida spp. and Cryptococcus neoformans strains, comprised those Fluconazole, Itraconazole and Genaconazole-resistant. A particularly significant antimycotic activity also resulted against Candida glabrata and Fusarium spp. strains, Itraconazole and Genaconazole-resistant, and against Candida krusei and Aspergillus fiimigatus, Fluconazole and Genaconazole-resistant, all pathogenic strains responsible for infections in immunodepressed patients.

The in vivo antimycotic activity has been evaluated, by intraperitoneal and by oral route, in a Candida model experimentally induced in mouse, against Candida albicans strains sensible to Fluconazole and Itraconazole.

From the in vivo experiments, the protective dose 50% (PD50) of the compounds of formula

II has been also determined and it resulted to be at least comparable to that of the reference compounds.

The compounds of formula π, object of the present invention, are therefore active on wide- range deep mycosis, but in particular against opportunistic pathogens responsible for infec¬ tions in immunodepressed patients, administrable by topic, oral and parenteral route and en-

dowed with a good therapeutic index.

The compounds of formula II are then useful in human or veterinary therapy for the treatment and prophylaxis of systemic and mucosal infections caused by fungi and yeasts. The significant pharmacological activity of the conφounds of formula II which, as already underlined, also results against strains resistant to antimycotic used in therapy and against recently isolated strains responsible for infections in immunodepressed patients, is particularly surprising in that such compounds are described in the European patent application No. 315946 as compounds for agriculture use only, having immunizing activity against fungal pathogenesis and phytoregulating activity on useful growings.

The pharmacological activity of the compounds of formula II is still more surprising :f it is considered that such compounds, while having some structural requirements common to the azole tertiary alcohols described in the literature, are diaracterized by the presence of a chain with two carbon atoms between the carbon atom bearing the hydroxy group and the oxygen or sulphur atom and by the presence of a polyfluorinated alkyl in the ether or thioether function. These structural requirements have never been described in the literature in combination -each other, as far as we know, within the field of compounds of the class of azole tertiary alcohols with antifungal activity. In this connection, the peculiarity of the structural requirements of the compounds of formula H, object of the present invention, is extremely surprising by considering that compounds having very similar structural requirements, such as the compound identified as RI and R2 namely 1 -( IH- 1 ,2,4-triazolyl)-2-hydroxy-2-(2,4-difluorophenyl)-3.3-dimethy l-4-ethoxybutane and l-(lH-l,2,-4-triazolyl)-2-hydroxy-2-(2,4-difluorophenyl)-3-( 2,2,3,3-tetrafluoropropoxy> butane, are substantially inactive in comparison with the compounds of formula II. For the human or v-eterinary use, the compounds of formula II can be administered in admix¬ ture with a suitable -carrier depending on the kind of administration route. Therefore, the pharmaceutical compositions containing a compound of formula II in admix¬ ture with a pharmaceutically acceptable carrier are a further object of the present invention. The compounds of formula II or their salts can be orally administered in the form of tablets, capsules, solutions or suspensions.

For the parenteral administration, for example by intravenous, intramuscular or subcutaneous

route, the compounds of formula II or their salts will be in the form of sterile aqueous solu¬ tions.

Alternatively, the compounds of formula II or their salts can be administered in the form of suppositories or pessaries.

For the topical administration the compounds of formula II or their salts will be preferably formulated as creams or powders.

For the oral or parenteral administration the daily dose of the compound of formula II will be generally from 0.1 to 50 mg/kg, preferably from 1 and 20 mg/kg, to be divided in one or more intervalled dos-es.

In order to better illustrate the present invention the following examples are now given.

Example 1

The following compounds were prepared according to the procedures described in the Euro¬ pean patent application No. 315946. r±Vl-flH-1.2.4-triazolylV2-hvdroxy-2-(2.4-did^oroDhenylι-3 -methyl-4-π.l.2.2-tetrafluoro- ethoxyibutane (Compound 1)

(±V 1 -( IH- 1.2.4-triazolyl V2-hvdroxy-2-( ^' 2.4-didιlorophenyl V3-methyl-4-( 1.1.2.2-tetrafluoro- ethoxy)hexane (Compound 2)

(± -l-nH-L2.4-triazolyl)-2-hvdroxy-2-.f4-chlorophenyl)-3.3-dime thyl-4-(1.1.2.2-tetrafluoro- ethoxylbutane (Compound 3)

( ±V 1 -fl H- 1 ■2.4-triazolylV2-hvdroxy-2-f 4-chlorophenyl ι-3-methyl-4-r 1.1.2.2-tetrafluoro- ethoxyibutane (Compound 4)

Example 2

Preparation of (±V 1 -fl H- 1 ■2.4-triazolylV2-hvdroxy-(2.4-difluorophenvI)-3.3-dimethyl -4- (1.1.2.2-tetrafluoroethoxy)butane (Compound 5)

Potassium hydroxide powder (5.33 mg; 9.50 mmoles) was added to a solution of (±V3-(2,4- difluorophenylV2,2-dimethyl-4-(lH-l,2,4-triazolyl)-l,3-butan ediol (5 g; 16.8 mmoles) and dimethylsulphoxide (8 ml) in toluene (60 ml), under stirring at -5°C.

The reaction atmosphere was substituted by tetrafluoroethylene and the mixture was kept un- der stirring at -5°C for 90 minutes.

After addition of water (120 ml), the organic phase was washed with 5% hydrochloric add

(80 ml) and treated with anhydrous sodium bicarbonate (6.5 g) under stirring for 30 minutes.

The liquid phase was filtered and the solvent was evaporated under reduced pressure to give an oil residue which was purified by flash d romatography (elueπt hexane:ethyi acetate=6:4). Compound 5 (5.7 g; 85% yield) was obtained as a white solid. m.p. 81-82.5°C

1H-NMR (CDC1 ₃ ): 8.01 (d, IH, CH-tetr.); 7.69 (s, IH, CH-cat ); 7.63-6.56 (m, 3H, Ar);

5.70 (tt, IH, JHF=53.4 Hz, CHF ₂ ); 5.33 (bs, IH, OH); 5.29-4.41 (m, 2H, CH ₂ -triazole), AB system: VA=4.19, VB=3.75, JAB=9.8 Hz, CH ₂ O; 1.06 (d, 3H, JHF=2.4 Hz, CH ₃ ); 0.98 (s, 3H, CH ₃ ).

By working in a similar way the following compounds were prepared.

(±Vl-(lH-1.2.4-triazolylV2-hvdroxy-2-(2.4-difluorophenyl V3-methyl-4-fl.l.2.2-tetrafluoro- ethoxytøutane (Compound 6) - 88% yield m.p. 112-114°C -NMR (CDCI3): 7.81 and 7.74 (2s, 2H, tetr.); 7.36-6.64 (m, 3H, Ar); 5.73 (tt, IH,

JHF=52.9 Hz, CHF ₂ ); 4.99 (bs, IH, OH); 4.95-4.53 ( , 2H, CH ₂ -triazole), AB portion of an

ABX system: VA=4.46, VB=3.94, JAB=10.2 Hz, JAX=7.3 Hz, JBX=4.9 Hz, CH ₂ O; 2.63-

2.45 (m, IH, CHCH3); 0.75 (d, 3H, CH3CH).

(±V l-flH- 1.2.4-triazolyl 2-hvdroxy-2-(2.4-difluorophenyl )-3-ethyl-4-( 1.1.2.2-tetrafluoroeth- oxy ibutane nitrate (Coπφound 7) - 80% yield m.p. l-44-145°C (acaetonitrile - isopropylether)

1H-NMR (CDCI3): 9.58 (s, IH, CH-triazole); 8.05 (s, IH, CH-triazole); 7.37-6.67 (m, 3H,

Ar); 6.08 (bs, H+); 5.80 (tt, IH, JHF=52.6 Hz, CHF ₂ ), AB system: VA=5.11, VB=4.87,

JAB=14.2 Hz, CH ₂ -triazole; AB portion of an ABX system: VA=4.42, VB=4.21, JAB=11.1 Hz, JAX=7.7 Hz, JBX=2.6 Hz, CH ₂ O; 2.41-2.30 (m, IH, CHCH ₂ ); 1.32-1.18 (m, 2H, CH-

CH ₂ ); 0.83 (t, 3H, JHH=7.1 Hz, CH ₃ ).

(±Vl-(lH-1.2.4-triazolylV2-hvdroxy-2-(2.4-dichlorophenyl V3.3-dimethyl-4-(1.1.2.2-tetra- fluoroethoxy)butane (Compound 8) - 67% yi-eld m.p. 102.5-104.5°C (hexane - isopropylether) 1H-NMR (CDCI3): 8.23 and 7.71 (2s, 2H, triazole); 7.86-7.12 (m, 3H, Ar); 5.57 (tt, IH,

JHF=53 Hz, CHF ₂ ); 5.86 (s, IH, OH); AB system: VA=5.86, VB=4.45, JAB=14.3 Hz,

CH ₂ -triazole; AB system: VA=4.23, VB=3.80, JAB=9.8 Hz, CH ₂ O; 1.13 and 1.02 (2s, 6H,

2CH ₃ ).

(±Vl-flH-1.2.4-triazolylV2-hvdroxy-2-f2.4-difluorophenyl V3.3-dimethyl-4-fl.1.2.3.3.3- hexafluoro- 1 -propyDbutane (Compound 9) - 18% yield by substituting tetrafluoroethylene with 1,1,2,,3,3,3,-hexafluoropropene. m.p. 84-85°C (hexane)

1H-NMR (CDC1 ₃ ): 8.00 (d, IH, CH-tetr.); 7.69 (s, IH, CH-cat.); 7.63-6.56 (m, 3H, Ar);

5.84 (bs, IH, OH); 5.78-4.38 (m, 2H, CH ₂ -triazole); 5.00-4.60 (m, IH, CHF); 4.35-3.73 (m, 2H, OCH ₂ ); 1.07- 1.00 (m, 6H, 2CH ₃ ).

Example 3

Preparation of 3-(2.4-difluorophenylV3.4-epoxy-2.2-dimethylbutanoic add -ethyl ester

A solution of isobutyric add ethyl ester (1.88 g; 16 mmoles) in tetrahydrofuran (10 ml) was added dropwise to a solution of lithium diisopropyiamide (prepared from n.butyllithium, 6.8 ml, 2.5 M solution in hexane and diisopropylamine, 2.38 ml, 1.72 g, 17 mmoles) in tetrahy¬ drofuran (10 ml) at -78°C.

After 30 minutes, a solution of l-(2,4-difluorophenyl)-2-bromoethanone (4.0 g; 17 mmoles) in tetrahydrofuran (10 ml) was added dropwise by keeping the temperature below -70°C.

The reaction mixture was kept under stirring for 90 minutes and the temperature was allowed to rise up to -30°C.

A saturated aqueous solution of ammonium chloride was added, the phases were separated and the aqueous phase was extracted with ethyl a-cetate (2 x 25 ml).

The collected organic phases were washed with an aqueous solution of sodium bicarbonate

5% (25 ml) and with water (25 ml), dried on sodium sulphate and evaporated under reduced pressure to give 3-(2,4-difluorophenylV3,4-epoxy-2,2-dimethylbutanoic add ethyl ester (4.2 g; 96% yield) as a light yellow oil.

1H-NMR (CDCI3): 7.41-6.67 (m, 3H, Ar); 4.11 (q, 2H, COOCH ₂ ); 3.26-2.71 (m, 2H,

CH ₂ O); 1.22 (t, 3H, CH ₃ CH ₂ ); 1.20 (s, 6H, 2CH ₃ ).

Example 4 Preparation of 3-(2.4-difluorophenyl)-3.4-epoxy-2.2-dimethyl-l-butanol

Lithium boron hydride (120 mg; 5.5 mmoles) was added to a solution of 3-(2,4-difluorophen-

ylV3,4-epoxy-2,2-dimethylbutanoic add ethyl ester (lg; 3.7 mmoles), prepared as described in example 3, in ethyl ether (10 ml).

After cooling at +10°C, methanol (0.22 ml; 3.5 mmoles) was added dropwise under stirring and the reaction mixture was kept at + 10°C under stirring for 2 hours.

Then, a 5% hydrochloric add solution (15 ml) was added dropwise, the phases were separated and the aqueous phase was extracted with ethyl acetate (2 x 15 ml).

The collected organic phases were washed with a 5% aqueous solution of sodium bicarbonate

(20 ml) and with brine (20 ml), dried on sodium sulphate and evaporated under reduced pres- sure to give an oil residue (0.8 g) which was purified by column chromatography on siiica gel

(eluent hexane: ethyl acetate=8:2) obtaining 3-(2,4-difluorophenyl)-3,4-q)oxy-2,2-dimethyl-l- butanol (823 g; 65% yield) as a light yellow oil.

1H-NMR (CDC1 ₃ ): 7.47-6.70 (m, 3H, Ar); 3.50-3.29 (m, 2H, CH ₂ OH); 3.31-2.72 (m, 2H,

CH ₂ O); 1.00 (s, 3H, CH3C); 0.93 (s, 3H, CH3C). Example 5

Preparation of 3-(2.4-difluorophenyl)-3.4-epoxy-2.2-dimethyl-l-butanol trifluoromethylsul- phonyl ester

Trifluoromethanesulphonic .anhydride (22 ml; 131 mmoles) was added dropwise to a solution of 3-(2,4-difluorophenylV3,4-epoxy-2,2-dimethyl-l-butanol (10 g; 44 mmoles), prepared as described in example 4, and pyridine (20.1 g; 263 mmoles) in methylene chloride (70 ml) at

-20°C, while keeping the temperature below -10°C.

The reaction mixture was kept under stirring at 0°C for 30 minutes, then a 10% aqueous solution of citric add (50 ml) was added.

The phases were separated, the aqueous phase was extracted with methylene chloride (50 ml) and the collected organic phase were washed with water (50 ml), dried on sodium sulphate and evaporated under reduced pressure at 15°C.

3-(2,4-difluorophenyl)-3,4-epoxy-2,2-dimethyl-l-butanol trifluoromethylsulphonyl ester (13 g;

81% yield) was obtained as a reddish oil which was used as such in the subsequent steps.

1H-NMR (CDCI3): 7.41-6.75 (m, 3H, Ar); 4.25 (s, 2H, CH ₂ OSO ₂ ); 3.19-2.71 (m, 2H, CH ₂ O); 1.07 (s, 6H, 2CH ₃ C).

Example 6

Preparation of 2-(2.4-difluorophenylr-2-r2-(2.2.3.3-tetrafluoroDroDoxyVl.l- dimethylethyl- oxirane

2,2,3, 3,-Tetrafluoropropanol (2.25 ml; 25 mmoles) was added dropwise to a suspension of sodium hydride (625 mg, 80%, 21 mmoles) in dimethyifoimamide (5 ml) at 0°C.

The mixture was kept under stirring at room temperature for 1 hour.

After cooling at -8°C, a solution of 3-(2,4-difluorophenyl)-3,-4-epoxy-2,2-<iimethyl-l-butanol trifluoromethylsulphonyl ester (3 g; 8.3 mmoles), prepared as described in example 5, in di¬ methyifoimamide (5 ml) was added. The reaction mixture was kept under stirring for 6 hours at 0°C, then poured into water (200 ml) and extracted with ethyl ether (4 x 25ml).

The organic phases were dried on sodium sulphate, evaporated under reduced pressure to give an oil residue (1.58 g) which was purified by column chromatography on silica gel (eluent hexane:ethyl acetate=99:l) obtaining 2-(2,4-difluorophenyIV2-[2-(2,2,3,3-tetrafluoroprop- oxy)-l,l-dimethylethyl]oxirane (1.2 g; 42% yield) as a colourless oil.

1H-NMR (CDC1 ₃ ): 7.42-6.70 (m, 3H, Ar); 5.88 (m, 2H, OCHF); 3.72 (m, 2H, OCH ₂ CF ₂ );

3.32-3.20 (m, 2H, CH ₂ OCH ₂ CF ₂ ); 3.12-2.64 (m, 2H, CH ₂ O); 0.98 and 0.97 (2s, 6H,

2CH ₃ C).

By working in a similar way, the following compound was prepared: 2-(2.4-difluorophenyl>-2-f2-(2.2.2-trifluoroethoxy)-l.l-d imethylethylloxirane as a colourless oil

44% yield

1H-NMR (CDCI3): 7.42-6.70 (m, 3H, Ar); 3.82-3.69 (m, 2H, OCH ₂ CF ₃ ); 3.41-3.25 (m,

2H, CH ₂ OCH ₂ CF ₃ ); 3.19-2.64 (m, 2H, CH ₂ O); 0.98 and 0.97 (2s, 6H, 2CH ₃ C). Example 7

Preparation of (±Vl-(lH-1.2.4-triazolyl)-2-hvdroxy-2-(2.4-difluorophenyl)- 3.3-dimethyl-4-

(2.2.3.3-tetrafluoropropoxy)butane (Compound 10)

A mixture of 2-(2,4-difluorophenyl)-2-[2-(2,2,3,3-tetrafluoropropoxy)-l,l -dimethyleth- yljoxirane (1.2 g; 3.5 mmoles), 1,2,4-triazole (480 mg; 7 mmoles) and potassium -carbonate (970 mg; 7 mmoles) in dimethyifoimamide (12 ml) was kept under stirring at 105°C for 8 hours.

The mixture was cooled, the predpitate was filtered and the solution was concentrate under reduced pressure.

The oil residue (1.5 g) was purified by column chromatography on silica gel (eluent hexane:ethyl acetate=8:2) to give compound 10 (1.1 g; 76% yield) as a white solid. m.p. 84-86°C -NMR (CDC1 ₃ ): 8.01 (bs, IH, triazole); 7.69 (s, IH, triazole); 7.62-7.65 (m, 3H, Ar);

5.87 (tt, IH, JHF=53.5 Hz, CHF ₂ ); 5.38-4.42 (m, 2H, CH ₂ -triazole); 5.22 (s, IH, OH);

3.88-3.75 (m, 2H, CH ₂ CF ₂ ); AB system: VA=3.63, VB=3.28, JAB=9.3 Hz, CH ₂ -O; 1.01- 0.96 (m, 6H, CH3CCH3).

By working in a similar way, the following compound was prepared:

(±)-l-(lH-1.2.4-triazolylV2-hvdroxy-2-(2.4-difluoropheny lV3.3-dimethyl-4-(2.2.2-trifluoro- ethoxyibutane (Compound 11) - 55% yidd m.p. 71-72°C 1H-NMR (CDCI3): 8.01-8.00 (m, IH, CH-triazole); 7.68 (s, IH, CH-triazole); 7.62-6.55 (m,

3H, Ar); 5.39-4.45 (m, 2H, CH ₂ -triazole); 5.23 (s, IH, OH); AB system: VA=3.84,

VB=3.77, JAB=8.7 Hz, CCH ₂ O; AB system: VA=3.70, VB=3.30, JAB=8.8 Hz, CH ₂ CF ₃ ;

1.00-0.96 (m, 6H, CH3CCH3).

Example 8 Preparation of O-r3-(2.4-difluorophenylV3-hvdroxy-2.2-dimethyl-4-(lH-1.2.4- triazolyl)l-but- yl-S-methyldithiocarbonate

A 50% aqueous solution of sodium hydroxide (25 ml) and tetrabutylammonium hydrogenc- sulphate (8 mg) were added to a solution of (±V3-(2,4-difluorophenylV2,2-dimethyl-4-(lH- l,2,4-triazoly)-l,3-butandiol (818 mg; 2.75 mmoles) in carbon disulphide (10 ml) and methyl- ene chloride (15 ml). Under stirring at 10°C, methyl iodide (430 mg; 3.02 mmoles) was added and the stirring was kept for 1 hour at 10°C.

The phases were separated, the aqueous phase was extracted with methylene chloride (25 ml), the collected organic phases were washed with water (25 ml), dried on sodium sulphate and evaporated to give a solid yellow residue (1.35 g) whidi was purified by column chromatography on silica gel (duent ethyl acetate:hexane=7:3) obtaining O-[3-(2,4- difluorophenylV3-hydroxy-2,2-dimethyl-4-(lH-l,2,4-triazolyl) ]butyl-S-methyldithiocarbonate

(950 mg; 89% yidd) as a light ydlow solid. m.p. 113-115°C

^Ϊ H-NMR (CDC1 ₃ ): 8.01 (d, IH, CH-tetr.); 7.70 (s, IH, CH-tetr ); 7.66-6.52 (m, 3H, Ar); 5.25-4.41 (m, 2H, CH ₂ -triazole); 4.71-4.40 (m, 2H, CH ₂ O); 2.55 (s, 3H, CH3S); 1.11 and

1.09 (2s, 6H, 2CH ₃ C).

Example 9

Preparation of (±Vl-(lH-1.2.4-triazolyl ^' )-2-hvdroxy-2-(2.4-difluorophenyl')-3.3-dimethyl-4- trifluoromethoxy-butane (Compound 12) The complex fluoridric add - pyridine [(HF)9/pyridine; 2.67 ml; 11.79 mmoles] and, then, O-

[3-(2,4-difluorophenylV3-hydroxy-2,2-dimethyl-4-(lH-l,2,4 -triazolyl)]butyl-S-methyldithio- carbonate (520 mg; 1.34 mmoles), prepared as described in example 8, dissolved in methylene chloride (2 ml) were added dropwise to a solution of l,3-dibromo-5,5-dimethylhydantoin (1.14 g; 4.02 mmoles) in methylene chloride (10 ml) at -70°C. The temperature was allowed to rise to -20°C and the rea-ction mixture was kept under stirring for 2 hours.

The mixture was poured into a 5% aqueous solution of sodium bisulphite (25 ml) and the mixture was brought to pH 14 with 33% sodium hydroxide.

The phases were separated, the aqueous phase was extracted with methylene chloride (25 ml) and the organic phases were dried on sodium sulphate, evaporated under reduced pressure and the oil residue (480 mg) was purified by column chromatography on silica gel (duent ethyl acetate:hexane=6:4) to give compound 12 (120 mg; 25% yield) as a white solid. m.p. 96-98°C

^-NMR (CDCI3): 8.02 (d, IH, JHH=2.9 Hz, CH-triazole); 7.70 (s, IH, CH-triazole); 7.63- 7.57 (m, 3H, Ar); 5.37 (s, IH, OH); AB portion of an ABX system: VA=5.26, VB=4.46,

JAB=13.96 Hz, JAX=3.0 Hz, JBX=2.2 Hz, CH ₂ -triazole; AB system: VA=4.20, VB=3.71,

JAB=9.5 Hz, CH ₂ OCF ₃ ; 1.09 (d, 3H, JHF=2.4 Hz, CH ₃ ); 0.98 (s, 3H, CH ₃ ).

Comparative Example A

Preparation of (±Vl-(lH-1.2.4-triazolylr-2-hvdroxy-2-(2.4-difluorophenyl)- 3.3-dimethyl-4- ethoxy-butane (Compound RI)

By working in a way similar to that described in example 6 but by using ethanol instead of

2,2,3,3-tetrafluoropropanol, the following compound was obtained.

2-(2,4-difluorophenyl 2-(2-ethoxy-l,l-dimethyl)ethyloxirane (20% yield) as a colourless oil. -NMR (CDC1 ₃ ): 7.45-6.70 (m, 3H, Ar); 3.38 (q, 2H, CH ₂ CH ₃ ); 3.20-2.60 (m, 2H, CH ₂ epoxide); 3.17-3.02 (m, 2H, CH ₂ OCH ₂ CH ₃ ); 0.98 (s, 3H, CH3C); 0.97 (s, 3H, CH3C).

By working in a way similar to that described in example 7, 2-(2,4-difluorophenyl)-2-(2- ethoxy-l,l-dimethyl)ethyloxirane was treated with 1,2,4-triazole obtaining compound RI as a white solid. 35% yidd m.p. 57-58°C

1H-NMR (CDCI3): 8.07 (bs, IH, CH-triazole); 7.68 (s, IH, CH-triazole); 7.60-6.57 ( , 3H, Ar); 5.47 (bs, IH, OH); 5.32-4.48 (m, 2H, CH ₂ -triazole); 3.45 (q, 2H, JHH=6.9 Hz, CH ₂ CH ₃ ); AB system: VA=3.30, VB=3.22, JAB=9.5 Hz, CH ₂ OCH ₂ CH ₃ ); 1.21 (t, 3H, CH ₃ CH ₂ ); 1 02 (d, 3H, J=2.3 Hz, CH3C); 0.94 (d, 3H, J=1.5 Hz, CH3C). Comparative Example B

Preparation of f±i-l-f lH-1.2.4-triazolylV2-hvdroxy-2-f2.4-difluorophenyl i-3-(2.2.3.3- tetrafluoropropoxVibutane (Compound R2)

2,2,3,3-Tetrafluoropropanol (2.25 ml; 25 mmoles) was added dropwise to a suspension of so¬ dium hydride (625 mg; 80%; 21 mmoles) in dimethyifoimamide (5 ml) at 0°C. The mixture was kept under stirring at room temperature for 1 hour, then 3-(2,4-difluoro phenyl)-4-(lH-l,2,4-triazolyl)-2,3-epoxy-butane (1.25 g; 5 mmoles), prepared as described in Bull. Chem. Soc. Jap., 67 (1994), 1427-1433, dissolved in dimethylformamide (1 ml) was added and the reaction mixture was kept at 105°C for 3 hours. The mixture was poured into water (50 ml) and extracted with ethyl ether (4 x 15 ml). The organic phases were dried on sodium sulphate, evaporated under reduced pressure to give -an oil residue (1.40 g) which was purified by column chromatography on silica gel (eluent ethyl acetate:hexane=6:4) obtaining compound R2 (385 mg; 20% yield) as a white solid, m.p. 77.0-78.5°C 1H-NMR (CDCI3): 7.85 and 7.72 (2s, 2H, 2CH-triazole); 7.48-6.66 (m, 3H, Ar); 5.91 (tt, IH, JHF=54.0 Hz, CHF ₂ ); 4.93-4.63 (m, 2H, CH ₂ -triazole); 4.31 (bs, IH, OH); 4.17-3.75 (m, 3H, CHOCH ₂ ); 1.05 (d, 3H, JHH=8.2 Hz, CH3CH)

Example 10 In vitro aπtimvcotic activity

The determination of the activity inhibiting the growth of mycetes was evaluated by the ma- cromethod of scalar brothdilutions in geometrical progression (M.R. McGinnis and M.G. Ri- naldi, "Antifungal drugs: mechanisms of action, drug resistance, susceptibility testing and as¬ say of activity in biological fluids", in aΛntibiotic in Laboratory Medicine, Ed. V. Lorian, Bal¬ timore 1991). As culture media Yeast Nitrogen Base broth (YNB) and Sabouraud Dextrose broth (SDB) were used for yeasts and moulds, respectively.

The results obtained in SDB (after incubation at 28°C for 7 days) were expressed as minimum inhibiting concentrations (MIC) the growth of my-cetes, while those obtained in YNB (after incubation at 35°C for 48 hours) were expressed as concentrations able to inhibit at 50% (IC50) the growth of the yeast. As reference compounds Fluconazole, Itraconazole and Genaconazole were used.

In the following tables the in vitro antimycotic activity data against various strains of Candida spp., Aspergillus spp., Fusarium spp., Cryptococcus neoformans and Trychop yton men- tagrophytes of the some representative compounds of formula II are reported.

Table 1 In vitro antimycotic activity of compounds 1-10, 12 and of the reference compound Flucona¬ zole against C. albicans, C. neoformans, T. mentagrophytes and A. fumigatus.

Compound IC ₅₀ (μg/ml) MIC (μg ml) C. albicans 1040 | C. neoformans T. mentagrophytes | A. fumigatus

1 0.0156 0.004 0.125 8

2 0.0312 0.125 2 4

3 0.0625 0.0625 1 >128

4 0.0312 0.5 4 >128

5 0.0156 0.0156 0.25 8

6 0.0312 0.0625 0.5 16

7 0.0312 0.125 0.5 16

8 0.0312 0.0156 0.25 >4

9 0.0156 0.125 2 >128

10 0.0156 0.0625 0.5 8

12 0.0156 0.0156 0.125 4

Fluconazole 0.5 2 16 >128

Table 2 In vitro antimycotic activity of compounds 1, 3, 5 and of the reference compounds Fluconazole, Itracona¬ zole and Genaconazole against Candida spp., Cryptococcus neoformans and other recently isolated strains (Trichosporon sp., B. capitatus, Fusarium sp.).

STRAINS (No.) Compound 1 Compound 3 Compound 5 Fluconazole Itraconazole Genaconazol MIC ₅₀ MICgo MIC ₅₀ MICgo MIC ₅₀ MICgo MIC ₅₀ MICgo MIC50 ^M,c 90 MIC ₅₀ MIC

C. albicans (7) ≤0.03 0.06 ≤0.03 2 ≤0.03 0.5 4 32 0.06 0.12 2

C spp ' (5) ≤0.03 ≤0.03 ≤0.03 ≤0.03 ≤0.03 ≤0.03 1 2 ≤0.03 0.06 0.5

C. tropicalis (2) 0.12 0.5 0.12 4-16 0.1 >64

C. krusei (2) ≤0.03-0.12 ≤0.03-0.5 ≤0.03 0.12 16->64 ≤0.03-0.12 16-32

C. glabrata (2) 0.12 0.5-2 0.12 32->64 2->16 32->64

C. neoformans (A) ≤0.03 ≤0.03 ≤0.03 0.5 ≤0.03 ≤0.03 1 32 ≤0.03 0.1 8

Trichosporon sp. (1) ≤0.03 0.12 ≤0.03 32 32

S. capitatus (1) ≤0.03 0.06 0.06 4 _ 16

Fusarium sp. (1) 0,25 16-64 0.25 — — 64

* C. parapsBosis, lusΛaniaβ, kefyr

Note to the table: MIC expressed as μg/ml

Within brackets the number of tested strains

Table 3 In vitro antimycotic activity of compound 1 and of the reference compounds Itraconazole and Genacona¬ zole against recently isolated Aspergillus niger and Fusarium spp.

Spedes Strain Compound 1 Itraconazole Genaconazole

A. niger 94-6222 ≤0.03 <0.03 32

Fusarium sp 81-256 0.25 2 64

Fusarium sp 86-3369 0.5 2-16 —

Fusarium sp 88-2116 0.5 2-8 —

Fusarium sp 946301 0.5 2-16 —

From the data reported in the table, it results that the compounds object of the present inven¬ tion have a wide-range in vitro activity. In particular, the conφounds of formula II are en¬ dowed with a significant activity also against Fluconazole and Itraconazole-resistant strains (Candida spp. and Cryptococcus neoformans) and against the other pathogenic strains re- sponsible for infections in immunodepressed patients (C. glabrata, C. krusei, Aspergillus spp. and Fusarium spp.).

Example 11 In vivo antimycotic activity Charles River white mice (CD1 strain) wdghing from 23 to 25 g, normo-feeded with standard diet and water ad libitum were used.

A suspension (0.2 ml containing 2.5x10' cdls/ml) of Candida albicans 1040 in physiologic solution was injected to each animal by venous route. Immediately after the infection and after 4, 24 and 48 hours, the animals received by oral route (in arable gum at 2%) doses increasing in a geometrical progression of the compound to be tested. The infected group was used as control.

The observation of the mortality of the mice was prolonged up to 14 days. From the number of survived animals for each concentration the average protective dose (DP50) was calculated by probits analysis (L.Lison - "Statistica applicata alia biologia sperimentale. La program- mazione deU'esperimento e l'analisi dd risultati" - Casa Editrice Ambrosiana, 1961). Compounds RI and R2 were used as comparative compounds. Fluconazole and Itraconazole were used as reference compounds.

The in vivo antimycotic activity data of some representative compounds of formula II after in vivo administration are reported in the following tables:

Table 4 In vivo antimycotic activity of compounds 1-3 and of the reference compound Itraconazole in mice experimentally infected by C. albicans, expressed as percentage of survival.

Compound dose mg/kg/os % percentage of survival after -day 3 6

1 25 100 100

2 25 100 100

3 25 100 100

Itraconazole 25 100 100

Control — 50 0

Table 5 Antimycotic efficacy by oral route of compounds 1, 3-7, 9, 12, RI, R2 and of the reference compounds Fluconazole and Itraconazole, expressed as average protective dose (PD50) after 8 and 14 days from the infection.

From the reported data it results that the compounds of formula π, object of the present in¬ vention, are active by oral administration at an average protective dose lower than that of Itraconazole and at least equivalent to that of Fluconazole.

Furthermore, the data reported in table 5 suggest, particularly for compound 1, characteristics similar to those of Fluconazole.