A method of treating fungal infections with a mixture of isomers of 2R,4S-hydroxyitraconazole and their sulfate and phosphate derivatives is disclosed. Pharmaceutical compositions containing these compounds are also disclosed.

FIELD OF THE INVENTION 
The present invention relates to a method of treating fungal infections 
employing any combination of two or more of the four isomers of 
2R,4S-hydroxyitraconazole, and phosphate and sulfate derivatives thereof. 
BACKGROUND OF THE INVENTION 
Itraconazole, a well-known antifungal agent, is defined in the USAN and USP 
Dictionary of Drug Names as 
4-[4-[4-[4-[[2-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1,3-di 
oxolan-4-yl]methoxy]phenyl]-1-piperazinyl]phenyl]-2,4-dihydro-2-(1-methylpr 
opyl)-3H-1,2,4-triazol-3-one or alternatively as 
(+)-1-sec-butyl-4-[p-[4-[p-[[(2R*,4S*)-2-(2,4-dichlorophenyl)-2-(1H-1,2,4- 
triazol-1-ylmethyl)-1,3-dioxolan-4-yl]methoxy]phenyl]-1-piperazinyl]phenyl] 
-.DELTA..sup.2 -1,2,4-triazolin-5-one. The commercially available material 
is the cis isomer in the dioxolane ring and is represented by the 
structural formula I: 
##STR1## 
It will be noted that there are three asymmetric carbons in formula I 
(denoted by asterisks): two in the dioxolane ring and one in the sec-butyl 
side chain on the triazolone. There are eight possible isomers of a 
structure having three asymmetric carbons: (R,R,R), (R,R,S), (R,S,S), 
(S,S,S), (R,S,R), (S,R,S), (S,R,R) and (S,S,R). Because the commercially 
available itraconazole is a cis isomer, it comprises a mixture of only 
those isomers that have a cis relationship in the dioxolane ring. Adopting 
the convention that the first denoted chiral center is at C-2 of the 
dioxolane ring, the second is at C-4 of the dioxolane and the third is in 
the sec-butyl group, commercial itraconazole is a mixture of (R,S,S), 
(R,S,R), (S,R,S) and (S,R,R) isomers. Compounds of this invention have the 
(2R,4S) configuration in the dioxolane ring and are substantially free of 
the SR isomers. 
The hydroxylation of the methylene carbon of the sec-butyl side chain 
creates an additional chiral center and gives rise to eight additional 
possible stereoisomers. The methods of the present invention employ a 
mixture of at least two of the four isomers of the two asymmetric centers 
in the butyl chain, i.e. a mixture of RSRR, RSRS, RSSR and RSSS; or a 
mixture of RSRR, RSRS and RSSR; or a mixture of RSRR and RSSR; etc. 
##STR2## 
The graphic representations of racemic, ambiscalemic and scalemic or 
enantiomerically pure compounds used herein are taken from Maehr J. Chem. 
Ed. 62, 114-120 (1985): solid and broken wedges are used to denote the 
absolute configuration of a chiral element; wavy lines indicate disavowal 
of any stereochemical implication which the bond it represents could 
generate; solid and broken bold lines are geometric descriptors indicating 
the relative configuration shown but denoting racemic character; and wedge 
outlines and dotted or broken lines denote enantiomerically pure compounds 
of indeterminate absolute configuration. Thus, among the structures below, 
those having open wedges are intended to encompass both of the pure 
enantiomers of that pair, those having solid wedges are intended to 
encompass the single, pure enantiomer having the absolute stereochemistry 
shown. 
Itraconazole is an orally active, broad-spectrum anti-fungal agent and is 
structurally related to miconazole and clotrimazole. It impairs the 
synthesis of ergosterol, which is the principal sterol of fungal cell 
membranes. This presumably results in an increased permeability and 
leakage of intracellular content. At high concentration, cellular internal 
organelles involute, peroxisomes increase, and necrotic changes occur. 
Following oral administration, itraconazole is slowly absorbed. Peak plasma 
levels are attained after 15 days of daily administration, and the 
pharmacokinetic behavior of itraconazole is nonlinear. The compound is 
eventually metabolized through the biologically active hydroxyitraconazole 
to several inactive metabolites. Metabolism is apparently through hepatic 
mechanisms, and in most subjects no metabolites are excreted in the urine 
[see, Hardin et al., Antimicro. Agents and Chemotherapy 32, 1310-1313 
(1988)]. 
The racemic mixture of itraconazole has been approved for use as an 
antifungal agent for blastomycosis and histoplasmosis. The compound is 
also being investigated for use in aspergillosis, coccidioidomycosis, 
cryptococcosis, onychomycosis, dermatophyte and candidiasis infections. 
Systemic fungal diseases (systemic mycoses) are usually chronic, very 
slowly developing conditions induced by opportunistic causative fungi 
which may not normally be pathogenic. However when they enter a host 
compromised by HIV, ionizing irradiation, corticosteroids, 
immunosuppressives, etc. or by such conditions as emphysema, 
bronchiectasis, diabetes mellitus, leukemia, burns and the like, they may 
become pathogenic. Symptoms in such fungal diseases may include fever, 
chills, anorexia and weight loss, malaise, and depression. Fungal diseases 
are often confined to typical anatomic distributions, and many involve a 
primary focus in the lung, with more characteristic manifestations of 
specific fungal infections when the fungus disseminates from a primary 
focus. For example, coccidioidomycosis occurs in a primary form as an 
acute, benign, self-limiting respiratory disease, with progressive disease 
developing from the primary form as a chronic, often fatal infection of 
the skin, lymph glands, spleen and liver. Similarly, blastomycosis 
primarily involves the lungs, and occasionally spreads to the skin. Other 
infectious diseases such as candidiasis and paracoccidioidomycosis offer a 
different course, and depending on the etiology may exhibit several forms 
involving the skin, mucous membranes, lymph nodes, and internal organs. 
Superficial fungal infections are caused by dermatophytes or fungi that 
involve the outer layers of the skin, hair or nails. The infections may 
result in a mild inflammation, and cause intermittent remissions and 
exacerbations of a gradually extending, scaling, raised lesion. Yeast 
infections including candidiasis, and oral candidiasis (thrush) are 
usually restricted to the skin, and mucous membranes, and the symptoms 
vary with the site of infection. 
Adverse effects associated with the administration of itraconazole include 
hepatotoxicity and inhibition of drug metabolism in the liver, leading to 
numerous, clinically significant, adverse drug interactions. [See, Gascon 
and Dayer Eur. J. Clin. Pharmacol. 41, 573-578 (1991) (interaction with 
midazolam); Honig et al. J. Clin. Pharmacol. 33, 1201-1206 (1993) 
(interaction with terfenadine); and Neuvonen et al. Clin. Pharmacol. 
Therap. 60, 54-61 (1996) (lovastatin).] Hypersensitivity reactions 
including urticaria and elevations in serum liver enzymes are also 
associated with the administration of the drug. Hepatoxicity is a less 
common but more serious adverse effect. Indeed, the use of oral conazoles 
as first line antifungals is usually discouraged because of the 
potentially serious consequences of the low incidence of hepatotoxicity 
[See, e.g., Lavrijsen et al., Lancet 340, 251-252 (1992)]. 
We have found evidence in our own studies in isolated guinea pig or rabbit 
hearts that the administration of racemic conazoles may be associated with 
an increased risk of cardiac arrhythmia. Arrhythmia has not been 
heretofore reported as a side effect of systemic itraconazole, although a 
particular subtype of arrhythmia, Torsades de Pointes, has been reported 
when racemic itraconazole was administered concurrently with terfenadine 
[Pohjola et al. Eur. J. Clin. Pharmacol. 45, 191-193 (1993)]. The lack of 
clinical reports of arrhythmia or QT anomalies may simply be a reflection 
of the fact that there is to date a relatively small subject population. 
The relative non-polarity and insolubility of itraconazole give rise to two 
other drawbacks: it cannot be readily formulated in parenteral solution 
and it does not penetrate the blood-brain barrier. As a result, numerous 
therapeutic indications which require rapid achievement of efficacious 
blood levels or access to the CNS are beyond treatment with itraconazole. 
In particular, central candidiasis, which may be responsible for AIDS 
related dementia, cannot be treated with itraconazole. 
Thus it would be particularly desirable to find a compound with the 
advantages of itraconazole which would not have the aforementioned 
disadvantages. 
SUMMARY OF THE INVENTION 
The various butyl chain isomers of 2R,4S hydroxyitraconazole and mixtures 
thereof possess potent activity in treating local and systemic fungal, 
yeast and dermatophyte infections while avoiding adverse effects 
associated with the administration of itraconazole. The butyl chain 
isomers of 2R,4S hydroxyitraconazole and mixtures thereof also enjoy the 
particular advantage of being more soluble in physiologically compatible 
solutions than is itraconazole. Moreover, we have found, quite 
surprisingly, that the side chain isomers of 2R,4S hydroxyitraconazole 
produce blood levels of hydroxyitraconazole that are significantly higher 
than the blood levels of hydroxyitraconazole and itraconazole that can be 
obtained from the administration of racemic [mixture of (R,S,S), (R,S,R), 
(S,R,S) and (S,R,R) isomers] itraconazole. 
In one aspect the invention relates to a method for treating fungal 
infection comprising administering to a mammal suffering from said fungal 
infection a therapeutically effective amount of a mixture of isomers of 
formulae: 
##STR3## 
wherein R is hydrogen, --P(O)(OH).sub.2, --SO.sub.3 H or a salt thereof. 
In another aspect, the invention relates to the mixture of isomers above. 
In another aspect the invention relates to pharmaceutical compositions 
comprising the mixture above and a pharmaceutically acceptable carrier. 
DETAILED DESCRIPTION OF THE INVENTION 
The four isomers cis-itraconazole (5-8) are shown below: 
##STR4## 
The eight isomers of cis-hydroxyitraconazole are shown below: 
##STR5## 
The isomers of cis hydroxyitraconazole have been made according to Scheme 1 
below in six forms: the pure R,S and S,R butyl chain isomers of 2R,4S and 
2S,4R and the R,R/S,S mixture of butyl chain isomers of each of the 2R,4S 
and 2S,4R dioxolanes. The synthetic approach is shown in detail for one 
enantiomer, 2 in Scheme 1; isomers 1, 9, and 10 may be made by the same 
synthesis employing other enantiomers of the starting materials. 
Similarly, the mixtures of RR/SS isomers in the butyl side chain, i.e. 
mixtures of 3/4 and 11/12, will be produced from the prochiral cis cyclic 
sulfate corresponding to the chiral trans cyclic sulfate 25. 
##STR6## 
As shown in Scheme 1, the chiral dioxolane (23) is prepared by a 
streospecific literature method from either R-3-tosyloxy-1,2-propanediol 
or S-3-tosyloxy-1,2-propanediol by acid-catalyzed ketalization to provide 
enantiomerically pure R,S- or S,R-(3) respectively. The dioxothiolane 25 
is prepared from a butanediol of appropriate configuration by treatment 
with thionyl chloride, followed by in situ oxidation of the resulting 
cyclic sulfite with sodium periodate in the presence of ruthenium 
trichloride. 
2,4-Dihydro-4-[4-[4-(4-methoxyphenyl)-piperazinyl]phenyl]-3H-1,2,4-triazol 
-3-one (26), prepared by the method of example XVII in U.S. Pat. No. 
4,267,179 (described below), is reacted with the dioxothiolane 25, 
prepared by the procedure of Gao and Sharpless [J. Am. Chem. Soc. 110, 
7538 (1988)], using potassium hydride in DMF in the presence of crown 
ether. The resulting methoxy-sulfate salt is cleaved to the phenol-alcohol 
28 by heating with 48% HBr at 100-110.degree. C. The tosyl ester 23 and 
the phenol-alcohol 28 are reacted in the presence of potassium hydroxide 
in DMF to provide the substantially enantiomerically pure product 2. 
Isomers 1, 9, 10, 3+4, and 11+12 are prepared in analogous fashion. Their 
rotations are shown in Table 1: 
TABLE 1 
______________________________________ 
Compound [.alpha..sub.D ].sup.25 .COPYRGT. = 0.1, MeOH) 
______________________________________ 
1 +12.7.degree. 
2 +22.3.degree. 
9 -22.0.degree. 
10 -10.6.degree. 
3 + 4(mix) +19.7.degree. 
11 + 12(mix) -18.5.degree. 
5 +19.2 
6 +14.0 
7 -13.4 
8 -18.7 
______________________________________ 
When individual isomers 3, 4, 11 and 12 are desired, they may be obtained 
by chromatography by methods well known in the art or by the methods 
described in PCT applications WO98/21204 and WO98/21205, the pertinent 
portions of which are laid out below. 
When it is desired to prepare the phosphate derivatives, 28 is treated 
first with t-butyldimethylsilyl chloride and diisopropylethylamine to 
protect the phenol, then with dibenzyl diisopropylphosphoramidite and 
t-butylhydroperooxide according to the procedure of PCT application 
WO95/17407, which is incorporated herein by reference, to phosphorylate 
the alcohol. The silyl protecting group is removed with anhydrous 
tetrabutylammonium fluoride and the benzyl-protected phosphate is coupled 
with the doxolane tosylate as in Scheme 1. The benzyl protecting groups 
are cleaved by hydrogenolysis in the presence of a palladium catalyst to 
provide the phosphate product. 
(2R,4S)-2-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)-4-tosyloxyme 
thyl-1,3-dioxolane tosylate DTTT (23 tosylate salt) is prepared as follows: 
A suspension of (R)-tosyloxy-1,2-propanediol (10.0 g, 40 mmol) and 
1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl) ethanone (10.0 g, 39 
mmol) in toluene (50 mL) is cooled to 5.degree. C. Triflic acid (15 mL, 4 
eq) is slowly added so that the temperature stays below 15.degree. C. 
After complete addition the reaction mixture (2 phases) is stirred at 
25.degree. C. for 60 h. The mixture is diluted with ethyl acetate (EtOAc) 
(200 mL) and slowly dropped into a solution of K.sub.2 CO.sub.3 (50 g) in 
water (400 mL) at 5.degree. C. The organic layer is separated and the 
aqueous layer washed with EtOAc (150 mL). The combined organic extracts 
are dried over Na.sub.2 SO.sub.4 (10 g) and filtered. A solution of 
toluenesulfonic acid (TsOH) (7.6 g monohydrate in EtOAc (50 mL) is slowly 
added at 25.degree. C. The white solid product is filtered after 30 min, 
washed and dried to give cis DTTT containing 5% trans. Two 
crystallyzations from CH.sub.3 CN (400 mL) gives 13.5 g pure (2R,4S)-DTTT 
(50% yield); [a].sub.D.sup.25 =+16.6.degree. (c=1, MeOH); ee=99.6%. The 
2S,4R isomer is prepared in analogous fashion from 
(S)-tosyloxy-1,2-propanediol (14.8 g, 60 mmol) and 
1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl) ethanone in (44% yield); 
[.alpha.].sub.D.sup.25 =-16.6.degree. (c=1, MeOH) ee=99.6% 
(4R,5R)-4,5-dimethyl-1,2,3-dioxathiolane 2,2-dioxide (25) is prepared as 
follows: A three-necked 500 mL round-bottom flask fitted with a reflux 
condenser and a calcium chloride drying tube was charged with 
(2R,3R)-(+)-2,3-butanediol (10.0 g, 10.1 ml, 0.11 mol) and carbon 
tetrachloride (120 mL). Thionyl chloride (16.0 g, 9.8 mL, 0.13 mol) was 
added dropwise at room temperature. Rapid gas evolution began. The 
reaction mixture was stirred at room temperature for 10 min, then warmed 
to reflux for 30 min to insure complete removal of HCl gas. The reaction 
mixture was cooled to 0.degree. C. in an ice-water bath and acetonitrile 
(120 mL), RuCl.sub.3 .cndot.H.sub.2 O (14 mg, 0.07 mmol), NaIO.sub.4 (35.6 
g, 0.17 mol) and water (180 mL) were added, respectively. The reaction 
mixture was allowed to warm to room temperature and stirred for 1.5 hr. 
The mixture was poured into methyl t-butyl ether (900 mL), and water was 
added to dissolve the remaining NaIO4 (ca. 600 mL). The phases were 
separated and the aqueous phase was extracted with methyl t-butyl ether 
(2.times.100 mL). The combined organic phases were washed with water 
(1.times.50 mL), saturated aqueous sodium bicarbonate (2.times.50 mL) and 
saturated aqueous bicarbonate (2.times.50 mL) and saturated aqueous sodium 
chloride (1.times.50 mL). The organic solution was dried over anhydrous 
magnesium sulfate and filtered through a bed of silica gel to give a clear 
and colorless solution. The solvent was removed in vacuo to give 16.01 g 
(95% yield) of the title compound. 
To a suspension of potassium hydride (530 mg, 4.6 mmol, 35 wt % dispersion 
in oil), prewashed with hexane (2.times.10 mL), in dimethylformamide (37 
mL) at room temperature was added 18-crown-6 (980 mg, 3.7 mmol) and 
2,4-dihydro-4-[4-[4-(4-methoxyphenyl)-1-piperazinyl]phenyl]-3H-1,2,4-triaz 
ol-3-one (6) (1.09 lg, 3.3 mmol). The solution was warmed to 80-85.degree. 
C. for 2 hr. then cooled in an ice-water bath to 0.degree. C. To this 
solution was added (4R,5R)-4,5-dimethyl-1,2,3-dioxathiolane 2,2-dioxide 
(5) (500 mg, 3.3 mmol). The reaction mixture warmed to 4.degree. C. After 
recooling to 0.degree. C., the reaction mixture was warmed to room 
temperature and stirred for 21 hours. To the reaction mixture was added 
150 mL of toluene and 400 mL of methyl t-butyl ether, and the white 
precipitated product was filtered from the mixture. The solid was dried in 
vacuo to give 1.70 g (quantitative yield) of a 87:13 mixture (by HPLC) of 
potassium 
(2R,3S)-3-[2,4-dihydro-4-[4-[4-(4-methoxyphenyl]-1-piperazinyl]phenyl]-3H- 
1,2,4-triazol-3-on-2-yl]but-2-yl sulfate (27). the title compound and 
triazolone starting material. A portion of this mixture was purified by 
flash chromatography (gradient from 95:5 chloroform:methanol to methanol) 
for characterization. 
To potassium 
(2R,3S)-3-[2,4-Dihydro-4-[4-[4-(4-methoxyphenyl-1-piperazinyl]phenyl]-3H-1 
,2,4-triazol-3-on-2-yl]but-2-yl[sulfate(7) (1.50 g, 2.98 mmol) and sodium 
sulfite (84 mg, 0.67 mmol) was added 48% HBr (6.0 mL). The solution was 
heated to 110-115.degree. C. for 7 hr and cooled to room temperature. The 
reaction mixture was poured into a wide-mouthed beaker and the pH raised 
to 7 by slow addition of solid potassium carbonate. Water was added (100 
mL), and the product was collected by filtration and dried in vacuo. Flash 
chromatography of the crude material eluting with a gradient of chloroform 
to 95:5 chloroform:methanol gave 1.03 g (70% yield) of 
2,4-dihydro-4-[4-[4-(4-hydroxyphenyl)-1-piperazinyl]phenyl]-2-[(1S,2R)-2-h 
ydroxy-1-methylpropyl)]-3H-1,2,4-triazol-3-one (28) as an adduct with 
SO.sub.3. 
To 
2,4-dihydro-4-[4-[4-(4-hydroxyphenyl)-1-piperazinyl]phenyl]-2-[(1S,2R)-(2- 
hydroxy-1-methylpropyl)]-3H-1,2,4-triazol-3-one SO.sub.3 adduct (28) (420 
mg, 0.86 mmol) and 
(1)--(2S,4S)-2-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl-4-tosyl 
oxymethyl-1,3-dioxolane tosylate (23 tosylate) (637 mg., 0.97 mmol) was 
added powdered potassium hydroxide (277 mg, 4.94 mmol) and 
N,N-dimethylformamide (15 mL). The reaction mixture was warmed to 
50-55.degree. C. for 8 hr and cooled to room temperature. Water was added 
(150 mL) and the crude product was collected by filtration and dried in 
vacuo. Purification by flash chromatography, eluting with ethyl acetate, 
followed by chloroform, 98:2.backslash.chloroform:methanol, then 95:5 
chloroform:methanol, gave 320 mg (52% yield) of 
(2S,4R)-4-[4-[4-[4-[[2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1 
,3-dioxolan-4-yl]methoxy]phenyl]-1-piperazinyl]-2,4-dihydro-2-[(1S,2R)-(2-h 
ydroxy-1-methylpropyl)]-3H-1,2,4,-triazol-3-one (2); [.alpha.].sub.D.sup.25 
=-22.0.degree. (c=0.1, MeOH). 
When the single isomers 3 and 4 are desired, they can be produced by two 
different modifications of Scheme 1, shown in Schemes 2 and 3: 
##STR7## 
2,4-Dihydro-4-[4-[4-(4-methoxyphenyl)-piperazinyl]phenyl]-3H-1,2,4-triazol- 
3-one (26) is reacted with the dioxothiolane 25 as described for Scheme 1, 
but the sulfate ester of the resulting methoxy-sulfate salt is hydrolyzed 
without cleaving the methyl from the phenol by heating with 48% HBr at 
45-50.degree. C. to produce alcohol 31. As shown in Scheme 2, alcohol 31 
may be treated with diisopropylazodicarboxylate, triphenylphosphine and 
benzoic acid according to the method of Mitsunobu [Synthesis 1981, 1-27], 
hydrolyzed with potassium hydroxide in methanol, and cleaved with 48% HBr 
at reflux to provide 33, the single R,R enantiomer analogous to 28. When 
the S,S enantiomer is desired, one begins with the opposite enantiomer of 
25. Alternatively, as shown in Scheme 3, the alcohol 31 may be treated 
with methanesulfonyl chloride in the presence of dimethylaminopyridine and 
diisopropylethylamine and the resulting mesylate ester of 34 inverted by 
treatment with cesium propionate according to the method of Senanayake et 
al. [Tet. Lett. 34, 2425 (1993)] to provide the propionate ester 36. The 
methoxy-ester 36 may be cleaved as before with potassium hydroxide and 
then HBr to give 33. 
Microbiological and pharmacologic studies can be used to determine the 
relative potency and the profile of specificity of the optically pure 
enantiomers, and the racemic mixture of itraconazole as antimycotic agents 
with a broad spectrum of activity against many fungi, yeast, and 
dermatophytes. 
With respect to antimicrobial activity of the aforementioned compounds, 
selected experiments are illustrated to profile useful antimicrobial 
activity, and not to limit this invention in any way, including the scope 
of susceptible microorganisms. Antifungal conazoles may be evaluated in 
vitro at several concentrations (in .mu.g/mL) against a number of fungi 
and bacteria. [See, Van Cutsem Chemotherapy 38 Suppl 1, 3-11 (1992) and 
Van Cutsem et al. Rev. Infec. Dis. 9 Suppl 1, S15-S32 (1987)]. The 
fungistatic assay is carried out in Sabouraud's liquid (1 g of neopeptone 
Difco and 2 g of glucose Difco per 100 mL of distilled water) in 
16.times.160 mm test tubes, each containing 4.5 mL of liquid medium which 
has been autoclaved at 120.degree. for 5 min. The compounds to be tested 
are dissolved in 50% alcohol at initial concentration of 20 mg/mL. The 
solutions are subsequently diluted with sterile distilled water to give a 
concentration of 10 mg/mL. Successive decimal dilutions are made in 
distilled water. To tubes containing 4.5 mL of Sabouraud's liquid medium 
0.5 mL of the solution of the drug is added, thereby obtaining 
concentrations of 1000, 500, 100, 10, and 1 .mu.g/mL of medium. Control 
tubes are prepared by adding 0.5 mL of distilled water to 4.5 of mL 
medium, alcohol being added to give concentrations identical with the 
tubes containing 1000 and 500 .mu.g of the drug. The filamentous fungi are 
incubated in Sabouraud's agar at 25.degree. for 2-3 weeks. A block of 
2.times.2.times.2 mm is then inoculated into the medium. All cultures are 
made in duplicate and are incubated at 25.degree. for 14 days. 
Itraconazole antifungal activity is enhanced in vitro in Sabouraud broth 
containing 10% inactivated bovine serum, and depends on the test medium 
used. Complete or marked inhibition of growth in Sabouraud broth after 14 
days of incubation may be observed with Microsporum canis, Trichophyton 
mentagrophytes, Candida albicans, Sporothrix schenckii, Paracoccidioides 
brasiliensis, Blastomyces dermatitides, Histoplasma spp., Aspergillus spp. 
and other fungi and bacteria. 
The hydroxyitraconazole isomers were tested for biological activity 
according three different assays. Table 2 summarizes the results of 
Kirby-Bauer Testing against three actively growing cultures: Candida 
albicans, Cryptococcus neoformens and Saccromyces cerevisiae. When 
compared to itraconazole, the zones of inhibition indicated that several 
hydroxyitraconazole isomers showed significantly greater potency. 
TABLE 2 
______________________________________ 
Kirby-Bauer Test (zones of inhibition in mm) 
Candida Cryptococcus 
Aspergillis 
Sacchromyces 
Compound albicans 
neoformens 
cerevisae 
______________________________________ 
1 22 28 27 22 
2 20 
9 20 
10 22 
3 + 4 23 
11 + 12 25 
5 + 6 + 7 + 8 
______________________________________ 
15 
Kirby-Bauer Testing 
A 10 mL tube containing 4 mL of Sabourauds dextrose broth was inoculated 
with 1 colony of Candida albicans picked from a plate of pure culture. The 
strain was ordered from the American Type Culture Collection (ATCC). The 
organism was grown for 4 hours at 30.degree. C. while shaking at 150 RPM. 
While the organism was growing, samples of hydroxy-itraconazoles were 
solubilized to a concentration of 10 mg/mL in DMSO. Each sample was then 
diluted 1:10 to make 1 mg/mL samples or 1000 .mu.g/mL. These samples were 
then diluted by serial 2-fold dilutions to produce samples now containing 
1000, 500, 250, 125, 62.5, 31.25, 15.6 .mu.g/mL. A 96-well microtiter dish 
was set up with 98 .mu.L of liquid growth media in each test well, along 
with 1 .mu.L of hydroxy-itraconazole solution. At 4 hours of growth time 
the Candida albicans was diluted to a 0.5 McFarland standard representing 
about 10.sup.5 -10.sup.6 cells/mL and 1 .mu.L of this inoculum placed into 
each test well of the microtiter dish. The dish was then covered and 
incubated at 30.degree. C. for 16 hours. 
Actively growing cultures of Candida albicans, Cryptococcus neoformens and 
Saccharomyces cerevisiae were prepared as described above. The cultures 
were diluted to a 0.5 McFarland standard and swabbed onto a 150 mm 
Sabouraud Dextrose agar plates. Paper disks (7 mm) were placed onto the 
agar plates using a disk dispenser. Next 10 .mu.l of 10 mg/mL solutions of 
each sample hydroxy-itraconazole was pipetted onto separate paper disks. 
The plates were then incubated at 30.degree. C. for 16 hours. Zones of 
inhibition were then measured in mm. 
Table 3 lists the MIC data for the hydroxyitraconazole isomers when tested 
against six strains of Candida albicans, and one strain each of Cr. 
neoformens, S. cerevisae and A. fumigatis. Nearly all the compounds showed 
good potency against Candida strains 28815 and 44203: 0.125 .mu.g/mL (0.17 
.mu.M) or lower. Good potency is also observed for A. fumigatis: 0.137 
.mu.g/ml (0.19 .mu.M). These numbers are compared to the MICs for 
itraconazole (Table 4) when tested against Candida albicans strain 44203. 
All the hydroxyitraconazole isomers were equipotent to the mixture of 
2S,4R-isomers of itraconazole (7 and 8) and more potent than the 
2R,4S-isomers (5 and 6). Moreover, the potencies of the itraconazole 
isomers were related to the stereochemistry of the dioxolane , whereas the 
antifungal potency did not appear related to the stereochemistry of the 
dioxolane ring in the hydroxyitraconazole isomers. 
TABLE 3 
______________________________________ 
Antifungal MICs (.mu.g/ml) 
Organism 1 2 9 10 3 + 4 11 + 12 
______________________________________ 
C. albicans strain 
10231 1.0 
0.5 
1.0 
28516 0.25 
0.25 
0.25 
0.25 
0.25 
28815 0.125 
0.5 
0.125 
0.125 
0.125 
44203 0.05 
0.05 
0.05 
0.025 
0.05 
44373 0.5 
0.25 
0.25 
0.25 
0.5 
62342 0.5 
1.0 
0.5 
Cr. neoformens 
0.39 
0.39 
0.78 
0.78 
0.78 
S. cerevisae 
0.195 
0.39 
0.195 
0.195 
0.195 
A. fumigatis 
0.137 
0.137 
0.137 
0.137 
______________________________________ 
0.137 
TABLE 4 
______________________________________ 
MICs Against Candida albicans Strain 44203. 
Isomer MIC .mu.g/ml 
______________________________________ 
1 0.05 
2 0.05 
9 0.05 
10 0.025 
3 + 4 0.10 
11 + 12 0.05 
5 0.3125 
6 0.3125 
7 &lt;0.078 
8 &lt;0.078 
5 + 6 0.156 
7 + 8 &lt;0.078 
5 + 6 + 7 + 8 0.06 
______________________________________ 
Table 5 gives IC.sub.50 data for the inhibition of cytochrome P450 3A4. The 
IC.sub.50 values were determined by the appearance of 
6.beta.-hydroxytestosterone after incubation with testosterone. All the 
compounds tested were significant inhibitors of testosterone 
6.beta.-hydroxylase activity. Isomers 1, 2 and 3+4, i.e. those with 2R,4S 
stereochemistry in the dioxolane ring, showed notably less inhibition of 
the P450 3A4 enzyme than the 2S,4R isomers. Of greater clinical 
significance is the ratio of the IC.sub.50 for 3A4 to the MIC for C. 
albicans 44203. This value indicates the "therapeutic index" for each 
isomer: a compound being a weak inhibitor of 3A4 (large IC.sub.50) and a 
potent antifungal against C. albicans 44203 (small MIC) would give a large 
IC.sub.50 /MIC ratio. As seen in Table 5, isomers 1, 2 and 3+4 showed 
excellent results. 
TABLE 5 
______________________________________ 
IC.sub.50 against Cytochrome P450 3A4 
IC.sub.50 /Itraconazole 
Compound IC.sub.50 (.mu.M) 
IC.sub.50 /MIC Ratio 
______________________________________ 
1 2.8 5.3 56 
2 70 
9 7.4 
10 19 
3 + 4 24 
11 + 12 6.5 
.+-.Itraconazole 
8.5 
______________________________________ 
In vivo activity of hydroxyitraconazole and derivatives may be compared 
against experimental cutaneous candidosis in guinea pigs, and vaginal 
candidosis in rats. The in vivo activity of the compounds in vaginal 
candidosis is evaluated by inducing vaginal infection with C. albicans in 
ovariectomized and hysterectomized Wistar rats (100 g) which are treated 
weekly with 100 .mu.g of estradiol undecanoate in sesame oil, 
subcutaneously. Animals in pseudooestrus are infected intravaginally with 
a fixed concentration of C. albicans in saline. Control of infection or 
cure is estimated by taking vaginal smears at fixed days after infection. 
Drugs to be evaluated and compared on a mg/kg basis may be given 
prophylactically or therapeutically and their efficacy judged by comparing 
the ratio of negative animals to the total number in each drug group. In 
similar studies, the activity against cutaneous candidosis in guinea pigs 
[(Van Cutsem et al. Chemotherapy 17, 392, (1972)] provides the basis of 
comparison. 
The compounds of the present invention allow the treatment of fungal 
infections while avoiding the adverse effects associated with 
itraconazole. The term "adverse effects" includes, but is not limited to, 
arrhythmogenicity, hepatotoxicity and elevations in serum liver enzymes, 
drug interactions, and hypersensitivity reactions including urticaria, 
nausea, vomiting, abdominal pain, headache, dizziness and the like. 
The potential for promoting arrhythmia is evaluated by examining the 
effects of the isomers of hydroxyitraconazole on cardiac action potential 
and contractility in human, canine and rabbit hearts. Torsades de Pointes 
is a well known side effect of antiarrhythmic drugs, such as quinidine, 
sotalol and acetyl-procainamide, which cause a prolongation of cardiac 
repolarization. All of these drugs have in common the ability to block a 
cellular potassium channel called the delayed rectifier (I.sub.K), and it 
is generally assumed that this is mechanistically linked to their ability 
to induce the syndrome of Torsades de Pointes. [See, Zehender et al. 
Cardiovascular Drugs Ther., 5 515-530 (1991).] Increases in QT duration 
and action potential duration in isolated guinea pig or rabbit hearts can 
therefore be used to indicate an arrhythmogenic effect. Hearts are 
perfused with an oxygenated Tyrode's solution, containing 0.0, 1.0, 5.0, 
10.0 or 30.0 .mu.M of racemic itraconazole. QT duration and action 
potential duration (APD) are measured from cardiac electrodes. 
To observe the effects in vivo, mongrel dogs of either sex weighing 5-20 kg 
are anesthetized and instrumented by standard techniques for blood 
pressure and EKG. A solid state transducer for dP/dT is placed in the left 
cardiac ventricle, and an epicardial electrode is put into place. The test 
compound is infused at progressively higher doses, beginning at 1 
.mu.g/kg/min for 15 to 30 minutes and increased incrementally until a 
cardiovascular collapse ensues. Parameters measured are: blood pressure, 
heart rate, dP/dT, and the QT-interval. Measurements of hemodynamics and 
electrical activity, including QT.sub.C interval, are made in response to 
the test compound and compared. 
The potential for promoting hepatotoxicity is assessed in vitro in human 
hepatic microsomes, human lymphocytes and other cell culture systems. 
Hepatic microsomes are prepared from human liver. Tissue is thawed and 
then homogenized in 0.15 M KCl in a Polytron homogenizer. The homogenate 
is centrifuged and the pellet is resuspended and homogenized in 0.15 M 
KCl. Aliquots are frozen and stored at -70.degree. C. Human lymphocytes 
are aseptically isolated from fresh, heparinized human blood. Blood is 
diluted with Eagle's minimal essential medium and layered on Ficoll-Paque. 
The samples are centrifuged, and lymphocytes are then removed from the 
aqueous-Ficoll interface and suspended in medium (15 Mm HEPES, pH, 7.4). 
The cells are then centrifuged, washed once in the HEPES medium, and 
resuspended. 
Cytotoxicity is assessed by the conversion of 3-(4,5 
dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to a purple 
formazan. The conversion of MTT to dye is done in multiwell plates. After 
preparation, hepatic microsomes or lymphocytes are incubated alone or with 
the test compound in a concentration range from 1 to 400 .mu.M at 
37.degree. C. in a humidified incubator. After incubation, the 
microsomes/cells are washed with 5% albumin in HEPES-buffered medium and 
resuspended. The microsomes/cells are then incubated at 37.degree. C. in a 
humidified incubator. After the incubation, 125 .mu.g of MTT is added to 
each well. The plates are incubated at 37.degree. C. and centrifuged. 
After centrifugation, 100 .mu.L of isopropanol is added and, after 
incubation, the optical density is determined using an automated 
plate-reader. 
The isomers of cis hydroxyitraconazole were tested for water solubility 
against a 0.5 M standard solution of hydroxyitraconazole. Table 6 lists 
the absorbance of each isomer at .lambda..sub.max =260 nm and its 
corresponding solubility. All the isomers except for 3+4 and 11+12 showed 
absorbances below the limit of detection (LOD) of the instrument (2 and 10 
were not even detected), allowing only order-of-magnitude accuracy. The 
values for 3+4 and 11+12 are barely above the LOD, giving accuracies of 
.+-.10%. In every case, solubilities were in the 10.sup.1 to 10.sup.2 
ng/ml range. Itraconazole itself has such low water solubility that it 
could not be determined by this method. 
TABLE 6 
______________________________________ 
Water solubility of Hydroxyitraconazole Isomers and Itraconazole. 
Compound Area (.mu.V*sec) 
Solubility (ng/ml) 
______________________________________ 
1 1499 51 
2 not detected 
9 45 
10 not detected 
3 + 4* 1970 
11 + 12* 590 
.+-.Itraconazole 
not detected 
______________________________________ 
LOD: 5000 .mu.V*sec. 
*Measured against a Hydroxyitraconazole std of 482 .mu.g/ml, Area = 
13364520 mV*sec 
The synthesis of the phosphate and sulfate esters of cis 
hydroxyitraconazole isomers and their corresponding MICs against three 
fungal strains are shown below. 
##STR8## 
TABLE 7 
______________________________________ 
Antifungal MICs (.mu.g/ml) 
Organism 2-sulfate 
2-phosphate 
______________________________________ 
C. albicans 6.25 12.5 
Cr. neoformens 12.5 
S. cerevisae 12.5 
______________________________________ 
The phosphate analogs of the isomers with the largest IC.sub.50 /MIC 
ratios, 1, 2 and 3+4 were further investigated for hydrolysis in plasma 
and tissues. Sodium, potassium and calcium salts were prepared, but the 
bis-NMG salt was the most stable. The A% conversion (as determined by HPLC 
A% analysis) of the phosphate ester to the hydroxy compound was determined 
in three systems: human plasma, equine pancreas and porcine brain. The 
results indicate that the phosphate ester of the 2R4SSR isomer 2 [both as 
the free acid and as the bis(N-methylglucamine) salt] was the most readily 
hydrolyzed compound in all three systems, and the bis-NMG salts were more 
or equally rapidly hydrolyzed when compared to the free acid counterparts. 
Hydrolysis proceeded to &gt;10% overnight at 37.degree. C. in pancreas and 
brain, and 3% in human plasma. The hydrolyses of the three phosphates of 
1, 2 and 3+4 were also studied in various rat tissues and plasma. In all 
cases, the jejunum and liver gave the greatest conversion to 
hydroxyitraconazole, ranging from 20% to 40% for the free acids. 
The water solubility of the NMG salts of the phosphate esters of isomers 1, 
2 and 3+4 were investigated and compared to the corresponding 
hydroxyitraconazole isomers 1, 2 and 3+4. The phosphate esters were as 
much as 10.sup.7 times more water soluble than the corresponding 
hydroxyitraconazole isomer. 
TABLE 8 
______________________________________ 
Water solubility of Hydroxyitraconazole Phosphate Isomers 
Compound Solubility (mg/mL) 
______________________________________ 
1 5.1 .times. 10.sup.-5 
2 &lt;1.0 .times. 10.sup.-5 
3 + 4 2.0 .times. 10.sup.-3 
1-PO.sub.3 --NMG.sub.2 
&gt;100 
2-PO.sub.3 --NMG.sub.2 
&gt;100 
3 + 4-PO.sub.3 --NMG.sub.2 
&gt;100 
______________________________________ 
The biological assays shown above establish that the hydroxyitraconazole 
isomers are of equal or greater potency than their itraconazole 
counterparts. All the hydroxyitraconazole isomers inhibited CYP3A4, their 
inhibition being a function of the absolute stereochemistry of the 
dioxolane moiety. In addition, the 2R,4S isomers of hydroxyitraconazole 
showed dramatically improved IC.sub.50 /MIC ratios when compared to 
racemic itraconazole. The sulfate and phosphate esters of 
hydroxyitraconazole isomers, although highly water-soluble, showed 
lessened antifungal activity. Nonetheless, the hydrolysis of the phosphate 
esters to hydroxyitraconazole allows them to be considered water-soluble 
prodrugs of useful potency. 
The magnitude of a prophylactic or therapeutic dose of hydroxyitraconazole 
or derivative in the acute or chronic management of disease will vary with 
the severity of the condition to be treated, and the route of 
administration. The dose, and perhaps the dose frequency, will also vary 
according to the age, body weight, and response of the individual patient. 
In general, the total daily dose range, for hydroxyitraconazole or a 
derivative, for the conditions described herein, is from about 50 mg to 
about 1200 mg, in single or divided doses. Preferably, a daily dose range 
should be between about 100 mg to about 800 mg, in single or divided 
doses, while most preferably, a daily dose range should be between about 
200 mg and 400 mg, in divided doses. In managing the patient, the therapy 
should be initiated at a lower dose, perhaps about 100 mg to about 200 mg, 
and increased up to about 400 mg or higher depending on the patient's 
global response. It is further recommended that children, and patients 
over 65 years, and those with impaired renal, or hepatic function, 
initially receive low doses, and that they be titrated based on individual 
response(s) and blood level(s). It may be necessary to use dosages outside 
these ranges in some cases as will be apparent to those skilled in the 
art. Further, it is noted that the clinician or treating physician will 
know how and when to interrupt, adjust, or terminate therapy in 
conjunction with individual patient response. An amount sufficient to 
alleviate or prevent infections but insufficient to cause adverse effects 
is encompassed by the above-described dosage amounts and dose frequency 
schedule. 
Any suitable route of administration may be employed for providing the 
patient with an effective dosage of hydroxyitraconazole or derivative. For 
example, oral, rectal, parenteral (subcutaneous, intramuscular, 
intravenous), transdermal, topical and like forms of administration may be 
employed. Dosage forms include tablets, troches, dispersions, suspensions, 
solutions, capsules, patches, ointments, creams, shampoos and the like. 
The pharmaceutical compositions of the present invention comprise 
hydroxyitraconazole or derivative as the active ingredient, or a 
pharmaceutically acceptable salt thereof, and may also contain a 
pharmaceutically acceptable carrier, and optionally, other therapeutic 
ingredients. 
The terms "pharmaceutically acceptable salts" or "a pharmaceutically 
acceptable salt thereof" refer to salts prepared from pharmaceutically 
acceptable non-toxic acids or bases including inorganic acids and bases 
and organic acids and bases. Since the hydroxy compound of the present 
invention is basic, salts may be prepared from pharmaceutically acceptable 
non-toxic acids including inorganic and organic acids. Suitable 
pharmaceutically acceptable acid addition salts for the compound of the 
present invention include acetic, benzenesulfonic (besylate), benzoic, 
camphorsulfonic, citric, ethenesulfonic, fumaric, gluconic, glutamic, 
hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, 
methanesulfonic (mesylate), mucic, nitric, pamoic, pantothenic, 
phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic, and the like. 
The phosphate, being acidic, allows for the preparation of salts of bases 
as well as internal salts. Suitable pharmaceutically acceptable base 
addition salts for the compounds of the present invention include metallic 
salts made from aluminum, calcium, lithium, magnesium, potassium, sodium 
and zinc or organic salts made from lysine, N,N'-dibenzylethylenediamine, 
chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine 
(N-methylglucamine) and procaine. N-methyl-glucamine salts are 
particularly preferred, inasmuch as they exhibit superior long term 
stability in parenteral and oral formulations. 
The compositions of the present invention include compositions such as 
suspensions, solutions, elixirs, aerosols, and solid dosage forms. 
Carriers such as starches, sugars, microcrystalline cellulose, diluents, 
granulating agents, lubricants, binders, disintegrating agents, and the 
like, are commonly used in the case of oral solid preparations (such as 
powders, capsules, and tablets), with the oral solid preparations being 
preferred over the oral liquid preparations. The most preferred oral solid 
preparation is tablets. 
Because of their ease of administration, tablets and capsules represent the 
most advantageous oral dosage unit form, in which case solid 
pharmaceutical carriers are employed. If desired, tablets may be coated by 
standard aqueous or nonaqueous techniques. 
A second preferred route of administration is topically, for which creams, 
ointments, shampoos, and the like are well suited. 
In addition to the common dosage forms set out above, the compounds of the 
present invention may also be administered by controlled release means 
and/or delivery devices and, because of their solubility, may also be 
employed in parenteral solutions, such as for intravenous administration. 
Because they reduce peak plasma concentrations, controlled release dosage 
forms are particularly useful for providing a therapeutic plasma 
concentration of 2R,4S-itraconazole while avoiding the side effects 
associated with peak plasma concentrations. 
Pharmaceutical compositions of the present invention suitable for oral 
administration may be presented as discrete units such as capsules, 
cachets, or tablets, or aerosol sprays, each containing a predetermined 
amount of the active ingredient, as a powder or granules, or as a solution 
or a suspension in an aqueous liquid, a non-aqueous liquid, an 
oil-in-water emulsion, or a water-in-oil liquid emulsion. Such 
compositions may be prepared by any of the methods of pharmacy, but all 
methods include the step of bringing into association the active 
ingredient with the carrier which constitutes one or more necessary 
ingredients. In general, the compositions are prepared by uniformly and 
intimately admixing the active ingredient with liquid carriers or finely 
divided solid carriers or both, and then, if necessary, shaping the 
product into the desired presentation. 
For example, a tablet may be prepared by compression or molding, 
optionally, with one or more accessory ingredients. Compressed tablets may 
be prepared by compressing in a suitable machine the active ingredient in 
a free-flowing form such as powder or granules, optionally mixed with a 
binder, lubricant, inert diluent, surface active or dispersing agent. 
Molded tablets may be made by molding in a suitable machine, a mixture of 
the powdered compound moistened with an inert liquid diluent. Desirably, 
each tablet contains from about 100 mg to about 300 mg of the active 
ingredient. Most preferably, the tablet, cachet or capsule contains either 
one of three dosages, about 50 mg, about 100 mg, or about 200 mg of the 
active ingredient. 
Similarly, sustained or controlled release formulation is well known in the 
art. Chapter 94 of a standard pharmacy school text, Remington: The Science 
and Practice of Pharmacy, 19th edition, entitled "Sustained-Release Drug 
Delivery Systems," describes the more common types of oral and parenteral 
controlled-release dosage forms (pages 1660-1675.) Controlled release 
means and delivery devices are also described in U.S. Pat. Nos. 3,845,770; 
3,916,899; 3,536,809; 3,598,123; and 4,008,719, and in PCT application WO 
92/20377. The relevant portions of the foregoing textbook and patent 
documents are incorporated herein by reference. 
For topical application, there are employed as non-sprayable forms, viscous 
to semi-solid or solid forms comprising a carrier compatible with topical 
application and having a dynamic viscosity preferably greater than water. 
Suitable formulations include but are not limited to solutions, 
suspensions, emulsions, creams, ointments, powders, liniments, salves, 
aerosols, etc., which are, if desired, sterilized or mixed with auxiliary 
agents, e.g., preservatives, stabilizers, wetting agents, buffers or salts 
for influencing osmotic pressure, etc. For topical application, also 
suitable are sprayable aerosol preparations wherein the active ingredient, 
preferably in combination with a solid or liquid inert carrier material, 
is packaged in a squeeze bottle or in admixture with a pressurized 
volatile, normally gaseous propellant, e.g., a freon. 
Formulations for parenteral administration include aqueous and non-aqueous 
sterile injection solutions which may contain anti-oxidants, buffers, 
bacteriostats and solutes which render the formulation isotonic with the 
blood of the intended recipient. Formulations for parenteral 
administration also include aqueous and non-aqueous sterile suspensions, 
which may include suspending agents and thickening agents. The 
formulations may be presented in unit-dose of multi-dose containers, for 
example sealed ampules and vials, and may be stored in a freeze-dried 
(lyophilized) condition requiring only the addition of a sterile liquid 
carrier, for example saline, phosphate-buffered saline (PBS) or the like, 
immediately prior to use. Extemporaneous injection solutions and 
suspensions may be prepared from sterile powders, granules and tablets of 
the kind previously described.

The invention is further defined by reference to the following examples 
describing in detail the preparation of the compositions of the present 
invention as well as their utility. It will be apparent to those skilled 
in the art that many modifications, both to materials and methods may be 
practiced without departing from the purpose and interest of this 
invention. 
EXAMPLE 1 
______________________________________ 
Oral Formulation - Capsules 
Quantity per capsule in mg 
Formula A B C 
______________________________________ 
Hydroxyitraconazole 
50 100 200 
Lactose 380 330 230 
Cornstarch 65 65 65 
Magnesium Stearate 
5 5 5 
Compression Weight 
500 500 500 
______________________________________ 
The active ingredient, hydroxyitraconazole or derivative, is sieved and 
blended with the excipients. The mixture is filled into suitably sized 
two-piece hard gelatin capsules using suitable machinery. Other doses may 
be prepared by altering the fill weight and if necessary, changing the 
capsule size to suit. 
EXAMPLE 2 
______________________________________ 
Oral Formulation - Tablets 
Quantity per tablet in mg 
Formula A B C 
______________________________________ 
Hydroxyitraconazole 
50 100 200 
Lactose 109 309 209 
Cornstarch 30 30 30 
Water (per thousand 
300 mL 300 mL 300 mL 
tabs)* 
Cornstarch 60 60 60 
Magnesium Stearate 
1 1 1 
Compression Weight 
250 500 500 
______________________________________ 
*The water evaporates during manufacture 
The active ingredient is blended with the lactose until a uniform blend is 
formed. The smaller quantity of cornstarch is blended with the water to 
form the resulting cornstarch paste. This is then mixed with the uniform 
blend until a uniform wet mass is formed and the remaining cornstarch is 
added and mixed until uniform granules are obtained. The granules are 
screened through a suitable milling machine using a 1/4" stainless steel 
screen. The milled granules are dried in a suitable drying oven and milled 
through a suitable milling machine again. The magnesium stearate is then 
blended and the resulting mixture is compressed into tablets of desired 
shape, thickness, hardness and disintegration. 
EXAMPLE 3 
______________________________________ 
Aqueous Suspension for Injection 
A suspending vehicle is prepared from the following materials: 
______________________________________ 
Polyethylene glycol 4000 
30 gm. 
Potassium chloride 11.2 gm. 
Polysorbate 80 2 gm. 
Methylparaben 0.2 gm. 
Water for injection q.s. 
1000 mL. 
______________________________________ 
The parabens are added to a major portion of the water and are dissolved 
therein by stirring and heating to 65.degree. C. The resulting solution is 
cooled to room temperature and the remainder of the ingredients are added 
and dissolved. The balance of the water to make up the required volume is 
then added and the solution sterilized by filtration. The sterile vehicle 
thus prepared is then mixed with 3 gm of 2R,4S-itraconazole or a phosphate 
salt thereof, which has been previously reduced to a particle size less 
than about 10 microns and sterilized with ethylene oxide gas. The mixture 
is passed through a sterilized colloid mill and filled under aseptic 
conditions into sterile containers which are then sealed. 
EXAMPLE 4 
______________________________________ 
Oral formulation - Controlled Release 
Composition per tablet: 
______________________________________ 
(+)-hydroxyitraconazole 50 mg 
lactose 100 mg 
dibasic calcium phosphate 
100 mg 
hydroxypropylmethylcellulose 
120 mg 
polyethylene oxide 100,000 MW 
20 mg 
polyethylene oxide 200,000 MW 
20 mg 
magnesium stearate 4 mg 
Total 414 mg 
______________________________________ 
EXAMPLE 4 
All of the ingredients, except the magnesium stearate, are blended for 10 
minutes. The mixture is screened through a 30-mesh (500 .mu.M) screen and 
reblended for a further 10 minutes. The magnesium stearate is screened 
through a 30-mesh (500 .mu.M) screen, added to the mixture and blended for 
five minutes. The resultant blend is made up into tablets, each weighing 
414 mg, on a rotary tableting machine. Tablets of other strengths may be 
prepared by altering the ratio of active ingredient to the excipients or 
to the final weight of the tablet.