EP2296644A1 - Combination compositions comprising antimycotic agents and statins and the use thereof - Google Patents

Combination compositions comprising antimycotic agents and statins and the use thereof

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Publication number
EP2296644A1
EP2296644A1 EP09742410A EP09742410A EP2296644A1 EP 2296644 A1 EP2296644 A1 EP 2296644A1 EP 09742410 A EP09742410 A EP 09742410A EP 09742410 A EP09742410 A EP 09742410A EP 2296644 A1 EP2296644 A1 EP 2296644A1
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day
spp
case
weight
candida
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German (de)
French (fr)
Inventor
Csaba VÁGVÖLGYI
Ildikó NYILASI
Tamás PAPP
Miklós PESTI
Gyöngyi LUKÁCS
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Pannonpharma Gyogyszergyarto Kft
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Pannonpharma Gyogyszergyarto Kft
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A61K31/405—Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics

Abstract

The present invention relates to a combination comprising one or more antimycotic agent(s) selected from the group of amphotericin B, nystatin, griseofulvin, primycin, terbinafine, an azole type one and one or more statin(s) selected from the group of lovastatin, simvastatin, fluvastatin, pravastatin, rozuvastatin, atorvastatin, cerivastatin, pitavastatin, for the use as an agent inhibiting the growth of fungi, furthermore, relates to a combination composition having antimycotic effect comprising a clinically acceptable dose of a combination of an antimycotic agent and a statin selected from the group of (i) amphotericin B and rozuvastatin, (ii) amphotericin B and pitavastatin, (iii) primycin and rozuvastatin, (iv) primycin and cerivastatin, (v) primycin and pitavastatin, (vi) voriconazole and simvastatin, (vii) voriconazole and pravastatin, (viii) voriconazole and atorvastatin, (ix) voriconazole and cerivastatin, and optionally one or more pharmaceutically acceptable excipients as well as relates to the said combination composition formulated as a medicine.˙

Description

Combination compositions comprising antimycotic agents and statins and the use thereof
The present invention relates to combinations comprising one or more active component(s) having antimycotic activity and one or more statin(s) or a prodrug or a metabolite thereof for use as agents inhibiting the growth of fungi.
More particularly, the present invention relates to combinations comprising (a) one or more active component(s) having antimycotic activity selected from the group consisting of amphotericin B, nystatin, griseofulvin, primycin, terbinafme, components of azole type, particularly ketoconazole, miconazole, fluconazole, itraconazole, clotrimazole, posaconazole, voriconazole, and prodrugs and metabolites thereof, and (b) one or more statin(s) selected from the group concicting of lovastatin, simvastatin, fluvastatin, pravastatin, rosuvastatin, atorvastatin, cerivastatin, pitavastatin, a prodrug and a metabolite thereof for the use as an agent inhibiting the growth of fungi, with the proviso that the combination is different from: amphotericin B and fluvastatin; ketoconazole and lovastatin; fluconazole and lovastatin; fluconazole and fluvastatin; fluconazole and pravastatin; itraconazole and fluvastatin; voriconazole and lovastatin; voriconazole and fluvastatin.
Furthermore, the present invention relates to combination compositions having antimycotic effect as well as to the use of these combinations in the therapy.
It is known that infections caused by pathogen fungi are spread worldwide, moreover, the number of severe infections is continuously increasing as a consequence of the growing number of patients with weakened immune system. Overcoming the infections is generally solved, but the antimycotic agents are, in many cases, effective only at very high doses. Increasing the concentration is, however, not only groundless and ineffective but at the same time deleterious. The reduced sensibilitiy (resistance) of several fungi against certain antimycotic agents may also constitute problems in the therapy. Therefore, there is a demand on medical solutions making possible to reduce the administered doses of the antimycotic agents or, at the same doses, to provide a more effective therapy. Attempts have been made to test new sorts of drugs as antimycotic agents and to combine antymicotic agents with other drugs, thereby to reduce the desired doses of the antimycotic agents by taking advantage of a possible synergic effect. The facts set forth below show that the attempts till now have attained only a limited success.
Lukacs el al. have studied the effect of lovastatin on species of Rhisomucor miehei and Rhisomucor piisillus [Journal of Clinical Microbiology, 2004, 42(1 1):5400-5402], They have found that lovastatin inhibited the growth of the said species, but the levels of inhibition were very different. The dose necessary for inhbiting the growth of Rhisomucor miehei was fifty- to sixty-fold higher as compared with that for the other species. Consequently, the inhibiting effect of lovastatin is different from species to species and depends on the circumstances of cultivation. The above reference does not mention any use of drug-combinations.
Roze et al. have disclosed that lovastatin, under certain circumstances of cultivation, caused apoptosis-like cell-death in the case of Mucor racemosiis [Fungal Genetics and Biology, 1998, 25(2): 1 19- 133]. This reference does not mention any combination.
Gyetvai et al. have disclosed that although lovastatin showed a fungistatic effect in the case of Candida albicans (i.e. inhibited the growth of the fungus), it did not exert an apoptotic effect against the said fungus [FEMS Yeast Research, 2006, 6(8):1 140-1148]. This reference does not mention any combination.
Macreadie et al. have studied antifungal activity of simvastatin and atorvastatin against several species of Candida {Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis) and Aspergillus fumigatus in 2006 [FEMS Microbiology Letters, 2007, 262:9-13]. Both simvastatin and atorvastatin, at certain concentrations, caused reduction and inhibition of growth, respectively, in the case of spp. Candida and Aspergillus fumigatus. The authors stated that inhibition of growth caused by statins could be ceased by adding argosterol or cholesterol. Neither this reference mentions any combination.
The above-mentioned studies have dealt with the antimycotic effect of statins without developing any combination.
Sud et al. have studied the effect of combinations of ketoconazole and other sterol-inhibitors (naphthalene, 15-azasterol, triamirol and lovastatin) on several fungi [Antimicrobial Agents and
Chemotherapy, 1985, 28(4):532-534]. Each studied sterol-inhibitor increased the sensibility more than fourfold against ketoconazole in certain fungi applied at concentrations lower than the minimal inhibitory concentration. Among the four compounds azasterol exerted the strongest effect by different ways in the case of each species of the studied fungi. The authors tried to prove their presumption that inhibitors of the sterol-synthesis combined with ketoconazole would significantly reduce the amount of ketoconazole necessary for antimycotic effect. They mentioned that certain combinations showed this effect at a clinically available dose
In contradiction to the above authors, Qiao et al. have found that neither lovastatin, nor simvastatin exerted, at clinically acceptable doses, any effect in vitro on the activity of itraconazole, voriconazole and amphotericin B against Aspergillus species [Medical Mycology, 2007, 45(7):589-593].
Chin et al. have studied the effect of several statins (fluvastatin, pravastatin, lovastatin, simvastatin) in the case of different Candida spp. (Candida albicans, Candida parapsilosis, Candida tropicalis) and Cryptococcus neoformans [Antimicrobial Agents and Chemotherapy, 1997, 41(4):850- 852], The said agents themselves exerted no (pravastatin, lovastatin and simvastatin) or very low antifungal activity, but in the case of the inhibition of growth exerted by the combinations fluvastatin/fluconazole and fluvastatin/itraconazole synergic and additive interactions could be shown. The concentrations of fluvastatin applied in the combinations were, however, much higher than the serum level maximally reachable in the blood, thus, this solution cannot be put into practice.
Nash et al. have studied the effect of the combinations fluvastatin/fluconazole and pravastatin/fluconazole in the case of Candida albicans, but in these studies fluvastatin and pravastatin were applied at clinically reachable concentrations [Journal of Medical Microbiology, 2002, 51(2): 105-
1099]. The authors did not found any synergic effect at these statin-concentrations, the observed level of reduction of growth was similar to that caused by fluconazole itself. Song et al. have studied the antifungal effect of the combination lovastatin/fluconazole and the effect thereof exerted on the expression of genes taking part in the sterol-synthesis [Medical Mycology, 2003, 41(5):417-25]. A synergic effect was demonstrated in vitro between lovastatin and fluconazole. The applied dose was, however, higher than that acceptable in the clinical practice (in vivo). Chamilos et al. have found lovastatin to be effective against a number of zygote fungi
(Ciinninghamella bertholletiae, Rhizopus homothalliciis, Rhi∑ppits ory∑ae, Mucor circinelloides) and they have shown a fungicide activity thereof being dependent on the culture medium and on the studied strains [Antimicrobial Agents and Chemotherapy, 2006, 50(l):96- 103]. The authors have demonstrated a significant synergic interaction in vitro in the case of lovastatin applied together with voriconazole which itself is ineffective. This was proved also in vivo using a Drosophila host model. The studied group of zygote fungi is, however, quite special and different from most of the pathogen fungi.
In the year 2007 Galgόczy et al. have studied the interaction between statins (lovastatin, simvastatin, rosuvastatin, atorvastatin) and an antifungal protein (PAF) produced by Penicillinum chrysogenum in the case of several zygote fungi (Rhizopus stolonifer, Mortierella wolfii, Syncephalastrum racemosum, Mycotypha africana) [FEMS Microbiology Letters, 2007, 270(l):109-l 15]. In a number of cases they have found a synergic interaction when applying the combinations lovastatin/PAF, simvastatin/PAF, rosuvastatin/PAF and atorvastatin/PAF. The studied groups of fungi are also of special nature, therefore, the results obtained in this study do not suggest that the solution could be operable also in the case of pathogen fungi. The used antifungal agent (PAF) is a protein of low molecular weight which is not used in the clinical practice and does not belong to the scope of substances studied in connection with the present invention.
Form the above facts one can come to the conclusion that, owing to the very different nature either of the statins or of the antimycotic agents or of the species of the studied fungi, it cannot be generally stated that the inhibitors of the sterol-synthesis combined with antimycotic agents show synergic effects. Lorenz et al. studied the effect of the combinations of lovastatin and 3 different antibiotics of azole type (ketoconazole, clotrimazole, miconazole) on the growth of Saccharomyces cerevisiae [Antimicrobial Agents and Chemotherapy, 1990, 34(9): 1660-1665]. The authors found that the minimal inhibitory concentration of the antifungal agents decreased to 1/6 to 1/32 value in the presence of lovastatin. The applied doses were, however, higher than the low levels acceptable in the clinical practice (in vivo). The studied species of fungus (baker's yeast) is not a pathogen but a species used as model-organism; therefore, the antimycotic effect found by the inventors of the present invention and claimed in this application is not deducible from the above results in general.
Natesan et al. have disclosed that fluvastatin enhanced the action of caspofungin in the case of Aspergillus fmnigatus but did not show any synergic intercation with voriconazole or amphotericin B [Diagnostic Microbiology and Infectious Disease, 2008, 60(4):369-73].
PCT publication No. WO2006013602 discloses the combined use of at least one statin, at least one omega-3 fatty acid and other substances for the treatment of cardiovascular diseases as well as diabetes, where the said substances show a synergic effect. PCT publication No. WO2006090756 relates to the combined use of phenylpropionic acid derivatives and statins for the treatment of obesity and diabetes, since the said substances together, by a synergic or additive effect, may lower the lipid concentration of the organism.
Patent application No. PCT/IB98/01220 relates to the combined use of amlodipine and statins for the treatment of arterosclerosis, hyperlipidaemia, angina pectoris and other cardiac diseases. The combinations of statins and amlodipine show additive or synergic effect in the treamtent of said diseases.
The above three documents do not mention the combination of the present invention or the antimycotic indication, either.
PCT publication No. WO9311777 discloses the combined use of amphotericin B and a glycerol ether for the treatment of fungal infection of mammals. By the use of glycerol ether the concentration of amphotericin B, necessary for reaching the efficiency, can be reduced; thus, these substances together show synergic interaction and inhibit particularly the growth of Cryptococcus neoformans and Candida albicans.
PCT publication No. WO03045319 discloses a therapeutic agent linked to a rifamycin derivative. The therapeutic agent may be, among others, a statin. Among the statins atorvastatin, rosuvastatin, lovastatin, simvastatin, pravastatin, cerivastatin and fulvastatin are mentioned and claimed particularly. The therapeutic agent is appropriate for preventing and treating microbial infections. Among the microbial infections, such as fungal infections, those caused by, among others, Candida spp. are disclosed and claimed. Neither of the last two documents discloses the increased antimycotic activity of the combinations or does not mention the combination of the present invention.
EP1631279 relates to a combination composition and a method for the treatment of neoplasms. The composition comprises a HMG-CoA reductase inhibitor and an azole compound. Among HMG-CoA inhibitors there are particularly mentioned and claimed, among others, simvastatin, lovastatin, pravastatin, fluvastatin, atorvastatin, cerivastatin, rosuvastatin and pitavastatin, whereas among the azole compounds there are particularly mentioned and claimed, among others, fluconazole, itraconazole, posaconazole, ketoconazole, clotrimazole and miconazole. Among neoplasms there are mentioned cancer of colon and lung, non small-cell lung carcinoma, cancer of ovary and prostate, and leukaemia.
PCT publication No. WO03086418 relates to a method for treating mammals afflicted by tumor/cancer by administering a polyene macrolide antibiotic and, preferably, an agent lowering the level of cholesterol. Among the polyene macrolide antibiotics there is particularly mentioned and claimed, among others, nystatin, whereas among the agents lowering the level of cholesterol there are particularly mentioned and claimed, among others, pravastatin, simvastatin, lovastatin, fluvastatin, cerivastatin and atorvastatin. The mentioned tumors/cancers are cancer of prostate, breast, ovary, tumor of kidney and epidermal carcinoma of mouth.
PCT publication No. WO07041677 discloses a device consisting of a combination of an implant and a drug which device inhibits scarring and formation of connective tissue between the implant and the tissues. Among the drug-combinations there are particularly mentioned and claimed, among others, the combination of lovastatin and itraconazole, furthermore in general, a combination of an azole compound with a HMG-CoA reductase inhibitor and, more generally, a combination of an antimycotic agent and a HMG-CoA reductase inhibitor.
Application No. US2005276836 discloses a vaginal device for delivery of therapeutic and sanitary substances. Among the substances used to coat the device there are mentioned, among many others, inhibitors of COXl and COX2, such as lovastatin, furthermore, as possible combination partners, amphothericin B, clotrimazole, fluconazole, itraconazole, ketoconazole, miconazole, nystatin and voriconazole. This application does, however, not mention either the fact that the said substances could be used as combinations having two members, or the enhanced antimycotic effect of the said combinations. PCT publication No. WO2006072881 discloses combination compositions comprising a therapeutically effective amount of the 2S,4R-enantiomer of ketoconazole and an agent lowering the level of cholesterol such as an HMG-CoA reductase inhibitor selected from the group of statins. Among HMG- CoA reductase inhibitors there are particularly mentioned statins, such as lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin and rosuvastatin, whereas among the indicated diseases there are mentioned, among others, hyperglycaemia, insulin resistance, lipidaemias, and several forms of diabetic retinopathy, nephropathy and neuropathy.
PCT publication No. WO2005023228 discloses a pharmaceutical formulation system enhancing the release of the substances having no or limited water-solubility into an aqueous environment. Among the acitve ingredients of the pharmaceutical formulation to be administered orally there are particularly mentioned and claimed, among others, amphotericin B, fluconazole, ketoconazole, griseofulvin, itraconazole, miconazole, terbinafine, atorvastatin, cerivastatin, fluvastatin, lovastatin, pravastatin, simvastatin and combinations thereof.
PCT publication No. WO2007143557 discloses a method for treating gastro-intestinal parasitic infections of mammals. The treatment comprises, among others, administration of a pharmaceutical composition. In the document there are mentioned, among others, inhibitors of HMG-CoA such as lovastatin; furthermore, the pharmaceutical composition may comprise, among others, antimycotic agents, such as fluconazole, itraconazole, ketoconazole, voriconazole, amphotericin B and nystatin.
PCT publication No. WO0048636 discloses pharmaceutical compositions comprising one or more active ingredient(s) and a special pharmaceutical component enhancing the absorption of the active ingredient(s), moreover phosphatidylcholine. Among the acitve ingredients there are mentioned, among others, antibiotics such as primycin, antimycotic agents such as amphotericin B, nystatin, griseofulvin, terbinafine, clotrimazole, ketoconazole, miconazole, fluconazole, itraconazole and HMG-CoA reductase inhibitors, such as lovastatin, pravastatin and simvastatin.
Published US application No. US20020071822 discloses a polymer for the treatment of Parkins.on's disease, wherein the polymer comprises, among others, one or more biologically active substance(s). The biologically active substances may be, among others, atorvastatin, pravastatin, fluvastatin, amphotericin B, primycin and nystatin.
EP 1275373 discloses a drug delivery system for avoiding interactions between several active ingredients and other contaminants that could negatively influence the release time and/or release site of the active ingredient. Among the active ingredients and contaminants there are particularly mentioned, among others, itraconazole, ketoconazole, lovastatin, fluvastatin, simvastatin, atorvastatin and combinations thereof. Published US application No. US200501 182272 relates to a fast releasing medicine or medicinal foodstuff comprising at least one active ingredient, at least one wetting agent and at least one solvent.
Among the active ingredients there are mentioned, among others, ketoconazole, miconazole, voriconazole, pitavastatin, rosuvastatin and combinations thereof.
Neither of the last three documents discloses any enhanced antimycotic effect of the combinations nor describes the combination of the present inventions.
When studying the state of the art disclosed in the scientific and patent literature, one can come to the following conclusions:
(1) there cannot be find any reference describing the generally valid antimycotic effect of the statins used by themselves; (2) no general synergic antimycotic interaction between the statins and the antimycotic agents can be detected; a synergic effect is incidental in the case of using combinations of a statin and an antimycotic agent: even in the case of active agents of similar stucture, the occurence of such an effect is accidental; according to the sate of art, one has not tried to demonstrate a synergic antimycotic effect: a number other indications are decribed for combinations falling under a scope that is the same as or similar to that of the present invention;
(3) antimycotic effect of combinations was tested only in the case of certain species, the presence or absence of a synergic effect was very different from species to species;
(4) the fact that a combination of a statin and an antimycotic agent shows in a special amount a synergic effect does not mean that the desired synergic interaction could be achieved also in the clinically acceptable dose ranges.
The aim of the present invention was to develop combination compositions the use of which at low doses effectively inhibits the growth of fungi, thereby making possible the reduction of the used amounts of the anitmycotic agents. The aim was, furthermore, to develop combinations wherein the similar solubility of the comprised agents enables the administration thereof in uniform pharmaceutical formulas to be applied externally or internally such as particularly in the form of tablets, capsules, creams, gels, ointments, powders, lacquers, solutions, foams etc.
The present invention is based on the recognition that certain combinations which have not been known till now show enhanced antimycotic effect. Furthermore, we have recognized that combinations described in the state of the art for other indications show enhanced antimycotic effect. The occurrence of a synergic interaction between the different active agents cannot be afore-seen on the basis of the known features of the different active agents. This is supported by the fact that certain combinations formed from active agents does not show such an advantageous activity as combinations formed from structurally similar other active agents. This fact can be concluded also from the documents forming the state of the art.
The invention is disclosed in details below. The terms set forth below are used in the present specification in the meanings as defined herein. If a technical term is not defined expressly, it is meant in the broadest sense generally known in that technical field. By the term "statin", as used in the present specification, it is meant a kind of medicine lowering the level of cholesterol the effect of which is based on the selective inhibition of 3-hydroxy-3-methyl- glutaryl coenzyme A (HMG-CoA) reductase enzyme. The enzyme catalyzes the conversion of HMG- CoA into mevalonic acid which step determines the speed of the isoprenoid biosynthetic pathway. Inhibition of the enzyme results in the early blocking of the cholesterol biosynthetic pathway. The term statin involves all structural and optical isomers of statins and all christal forms and modifications thereof, too.
By the terms "antimycotic agent" and "antifungal agent", which are equivalent as used in the present specification, it is meant a medicinal substance used for the treatment or prevention of local and/or systemic infections caused by fungi. The term antimycotic agent (and antifungal agent) involves all structural and optical isomers of the said compounds and all christal forms and modifications thereof, too.
By the terms "antimycotic agent of azole type" and "antifungal agent of azole type", which are equivalent as used in the present specification, it is meant a compound of natural or synthetic origin containing a five-membered aromatic heterocycle comprising at least one nitrogen, sulfur or oxygen atom beside carbon atoms and exerting an effective antimycotic (antifungal) activity. The antimycotic (antifungal) azole compounds inhibit the action of the enzyme citochrome P450 14α-demethylase which is an enzyme component of the biosynthetic pathway of ergosterol, a component of the cell-membrane of fungi. The term antimycotic agent (and antifungal agent) of azole type involves all structural and optical isomers of the said compounds and all christal forms and modifications thereof, too. By the term "dose", as used in the present specification, it is meant a portion, an amount of a medicine to be taken according to the physician's prescription, regardles of its expression form as unit. In the present specification and claims a dose may be expressed in the forms of mg/kg body weight pro day, mg/day, % by weight, % by volume, % by weight/volume etc.
By the term "serum level", as used in the present specification, it is meant a concentration of the substance in question in the blood-serum.
By the term "prodrug", as used in the present specification, it is meant a substance wherefrom another substance, an element of the synthetic pathway of the substance in question, is formed by chemical reactions which result in a stable end-product of the synthetic pathway,
By the term "metabolite", as used in the present specification, it is meant a substance taking part in the metabolism (decomposition), a substance taking part or being formed in the decomposition of complex organic compounds or in metabolic processes.
The meaning of the term "synergic" is in general sense: if a combination of two active components enhances the intensity or duration of the effects of the components by a greater extent than the algebraical sum of the separate effects. By the term "synergic", as used in the present specification, it is meant an interaction calculated by the mathematical relation (Abbott's formula) used for determinig the synergism:
^ 'obtained' ^anticipated
UcPaBd = X + Y - (XY/100) wherein X represents the percentage of inhibition caused by the statin itself and
Y represents the percentage of inhibition caused by the antifungal agent itself. If the obtained value is 0.5 < IR < 1.5, the interaction is additive; if IR > 1.5, the interaction is synergic, whereas if IR < 0.5, the interaction is antagonistic.
The meaning of the term "additive" is in general sense: if a combination of two active components enhances the intensity or duration of the effects of the components and this effect is equal to the algebraical sum of the separate effects. By the term "additive", as used in the present specification, it is meant an interaction calculated by the mathematical relation (Abbott's formula) used for determinig the synergism.
It is to be noted that, in the present technical field, the additive interaction itself can result in an unexpected increase of the effectiveness, therefore, the additive interaction as used in the present specification should not be confused with the common term of addition not inplicating any surplus effect. The present invention relates, in a first aspect, to a combination comprising one or more active component(s) having antimycotic activity [i.e. antimycotic agent(s)] and one or more statin(s), a prodrug or a metabolite thereof for the use as an agent inhibiting the growth of fungi. According to the present invention, the antimycotic agent may be a compound of azole type, without to be limited thereto. An antimycotic agent used in the present invention is preferably selected from the group of amphotericin B, nystatin, griseofulvin, primycin, ketoconazole, miconazole, fluconazole, itraconazole, clotrimazole, posaconazole, voriconazole, terbinafine, a prodrug thereof or a metabolite thereof. The statin used in the present invention is preferably selected from the group of lovastatin, simvastatin, fluvastatin, pravastatin, rosuvastatin, atorvastatin, cerivastatin, pitavastatin or a prodrug or a metabolite of the said compounds.
The combination of the present invention may comprise one or more whether of the antimycotic agent(s) or of the statin(s), with the proviso that the combination is different from the following ones: amphotericin B and fluvastatin; ketoconazole and lovastatin; fluconazole and lovastatin; fluconazole and fluvastatin; fluconazole and pravastatin; itraconazole and fluvastatin; voriconazole and lovastatin; voriconazole and fluvastatin.
The combination of the present invention, comprising one or more of antimycotic agent(s) and statin(s) or a prodrug or a metabolite of the said compounds, comprises the active components preferably in a clinically acceptable dose. The clinically acceptable daily dose, as used in the present specification and claims, for an adult person with a body weight of 65 to 70 kg is preferably
(i) in the case of amphotericin B if administered intravenously, 0.01 to 2 mg/kg body weight/day, if administered orally, 10 to 10,000 mg/day, if adminstered topically, a concentration of 0.001 to 5 % by weight; (ii) in the case of nystatin if administered orally or topically, an amount of 10,000 to 10,000,000 USP
Nystatin-Unit;
(iii) in the case of grisoufulvin if administered orally, 0.1 to 2000 mg/day, preferably 0.1 to 1000 mg/day, more preferably 0.1 to 250 mg/day;
(iv) in the case of primycin if administered topically, a concentration of 0.02 to 5 % by weight;
(v) in the case of terbinafine if administered orally, 0.1 to 400 mg/day, preferably 0.1 to 100 mg/day, more preferably 0.1 to 50 mg/day; if adminstered topically, a concentration of 0.001 to 5 % by weight; (vi) in the case of an antimycotic agent of azole type if administered orally, 0.1 to 1200 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if adminstered topically, a concentration of 0.001 to 5 % by weight; (vii) in the case of ketoconazole if administered orally, 0.1 to 1200 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if adminstered topically, a concentration of 0.001 to 5 % by weight;
(viii) in the case of miconazole if administered orally, 0.1 to 1000 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if adminstered topically, a concentration of 0.001 to 5 % by weight;
(ix) in the case of fluconazole if administered orally, 0.1 to 1200 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if adminstered topically, a concentration of 0.001 to 5 % by weight; (x) in the case of itraconazole if administered orally, 0.1 to 1200 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if administered intravenously, 0.01 to 20 mg/kg body weight/day; if adminstered topically, a concentration of 0.001 to 5 % by weight; (xi) in the case of clotrimazole if administered orally, 0.1 to 500 mg/day, preferably 0.1 to 250 mg/day, more preferably 0.1 to 100 mg/day; if adminstered topically, a concentration of 0.001 to 5 % by weight; (xii) in the case of posaconazole if administered orally, 0.1 to 800 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if adminstered topically, a concentration of 0.001 to 5 % by weight;
(xiii) in the case of voriconazole if administered orally, 0.1 to 400 mg/day, preferably 0.1 to 100 mg/day, more preferably 0.1 to 50 mg/day; if administered intavenously, 0.1 to 12 mg/kg body weight/day; if adminstered topically, a concentration of 0.001 to 5 % by weight;
(xiv) in the case of lovastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day;
(xv) in the case of simvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day; (xvi) in the case of fluvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 20 mg/day; (xvii) in the case of pravastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day;
(xviii) in the case of rosuvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 5 mg/day;
(xix) in the case of atorvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day; (xx) in the case of cerivastatin 0.025 to 5 mg/day, preferably 0.025 to 2.5 mg/day, more preferably 0.025 to 1 mg/day; (xxi) in the case of pitavastatin 0.1 to 80 mg/day, preferably 0.1 to 40 mg/day, more preferably 0.1 to 5 mg/day for the use as an agent inhibiting the growth of fungi.
However, it is not intended to limit the clinically acceptable dose to the above ranges. In a further aspect, the present invention relates to a combination composition having antimycotic effect comprising a combination of an active component having antimycotic activity and a statin, a prodrug or a metabolite of the said compounds selected from the goup consisting of (i) amphotericin B and rosuvastatin, (ii) amphotericin B and pitavastatin, (iii) primycin and rosuvastatin, (iv) primycin and cerivastatin, (v) primycin and pitavastatin, (vi) voriconazole and simvastatin, (vii) voriconazole and pravastatin, (viii) voriconazole and atorvastatin, (ix) voriconazole and cerivastatin, and optionally pharmaceutically acceptable excipients. The preferred composition of the invention comprises the active components in a clinically acceptable dose.
The clinically acceptable doses, as used in the present specification and claims, contain in the case of combination (i), 0.1 to 99 % by weight of amphotericin B and 0.1 to 99 % by weight of rosuvastatin; in the case of combination (ii), 0.1 to 99 % by weight of amphotericin B and 0.1 to 99 % by weight of pitavastatin, in the case of combination (iii), 0.1 to 99 % by weight of primycin and 0.1 to 99 % by weight of rosuvastatin, in the case of combination (iv), 0.1 to 99 % by weight of primycin and 0.1 to 99 % by weight of cerivastatin, in the case of combination (v), 0.1 to 99 % by weight of primycin and 0.1 to 99 % by weight of pitavastatin, in the case of combination (vi), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of simvastatin, in the case of combination (vii), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of pravastatin, in the case of combination (viii), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of atorvastatin, in the case of combination (ix), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of cerivastatin; however, it is not intended to limit the clinically acceptable dose to the above ranges.
The combinations and compositions of the present invention can be used against the following species of fungi: Candida spp. (such as Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida giiillermondii, Candida krusei and Candida kefyr), Cryptococcus spp. (such as Cryptococcus neoformans), Pneumocystis carinii, Aspergillus spp. (such as Aspergillus flavus, Aspergillus fiimigatus, Aspergillus terreus, Aspergillus repens, Aspergillus versicolor), Mucor spp. (such as Mucor circinelloides, Mucor indicus, Mucor ramosissimus), Rhizomucor spp. (such as Rhizomucor pusillus, Rhizomucor iniehei), Rhi∑opus spp. (such as Rhizopus oryzae, Rhizopus microsporus var. microporus, Rhizopus microsporus var. rhizopodiformis, Rhizopus schipperae), Absidia (such as Absidia corymbifera), Apophysomyces (such as Apophysomyces elegans), Cunninghamella (such as Cunninghamella bertholletiae), Saksenaea (such as Saksenaea vasiformis), Cokeromyces (such as Cokeromyces recurvatus), Syncephalastrum (such as Syncephalastrum racemosum), Basidiobolus (such as Basidiobolus ranarum), Conidiobolus (such as Conidiobolus coronatus), Coccidioides spp. (such as Coccidioides immitis), Paracoccidioides spp. (such as Paracoccidioides brasiliensis), Histoplasma spp. (such as Histoplasma capsulatum) or Blastomyces spp. (such as Blastomyces dermatitidis), Trichophyton spp. (such as Trichophyton mentagrophytes, Trichophyton rubrum), Microsporum spp. (such as Microsporum canis, Microsporum gypseum) or Epidermophyton spp. (such s Epidermophyton floccosum) and Pityrosporum spp. [such as Pityrosporum orbiculare (Malassezia furfur)], Geotrichum (such as Geotrichum clavatum, Geotrichum candidum), Trichosporon (such as Trichosporon beigelii), Blastoschi∑omyces (such as Blastoschizomyces capitatus), Sporothrix (such as Sporothrix schenckii), Scedosporium (such as Scedosporium apiosperum, Scedosporium proliβcans), Cladosporium (such as Cladosporium cladosporioides, Cladosporium sphaerospermum), Acremonium spp. (such as Acremonium falciforme), Beauveria spp. (such as Beauveria bassiana), Fusarium spp. (such as Fusarium oxysporum, Fusarium solani, Fusarium moniliforme), Paecilomyces spp. (such as Paecilomyces variotii, Paecilomyces lilacinus, Paecilomyces marquandiϊ), Penicillium spp. (such as Penicillium marneffei, Penicillium purpurogenum), Scopulaήopsis spp. (such as Scopulariopsis brevicaulis), Cladophialophora spp. (such as Cladophialophora bantiana, Cladophialophora carrionii), Aureobasidium spp. (such as Aureobasidium pullulans), Bipolaris spp. (such as Bipolaris australiensis, Bipolaris hawaiiensis, Bipolaris spicifera), Curvularia spp. (such as Curvularia lunata, Curvularia pallescens, Curvularia geniculata), Exophiala spp. (such as Exophiala spinifera), Exserohilum (such as Exserohilum rostratum), Phialophora spp. (such as Phialophora parasitica), Wangiella spp. (such as Wangiella dermatitidis), Alternaria spp. (such as Alternaria alternata), Chaetomiitm spp. (such as Chaetomium globosum), Coniophora spp. (such as Coniophora puteana). The combinations and compositions according to the present invention can be used preferably against the following species of fungi: Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida giiillermondii, Candida krusei, Candida kefyr, Aspergillus flavus, Aspergillus fiimigatus, Rhizopus oryzae, Trichophyton mentagrophytes, Trichophyton rubrum, Microspomm canis, Microsponim gypseiim, Pityrosponim orbicitlare.
In a further aspect, the present invention relates to the combination composition of the ionvention formulated as a medicine. The medicine may be in a form to be administered externally or internally, such as particularly, but not limited to, a tablet, capsule, cream, gel, ointment, powder, lacquer, solution, foam. In a further aspect, the present invention relates to the combination composition of the invention for the use as medicine, especially as a medicine having antimycotic effect.
Finally, the present invention relates to the use of the combinations of active components of the invention for preparing compositions having enhanced antimycotic effect.
Although it is not intended to limit the explanation of the present invention to a special theory, it is presumed that the unexpected recognition which is based the present invention on may rest upon the following mechanism of action. The action of statins is based upon the selective inhibition of 3-hydroxy- 3-methylglutaryl-coenzyme A (HMG-CoA) reductase enzyme. The enzyme catalyzes the conversion of HMG-CoA into mevalonic acid which step determines the speed of the isoprenoid biosynthetic pathway, thus the statins inhibit the biosynthesis of sterols. Since ergosterol, also a compound of isoprenoid type, is one of the components of the cell membrane of fungi, statins are effective in inhibition of the growth of fungi. The antimycotic agents, exploiting the difference between the human cells and the cells of fungi, selectively inhibit the proliferation of fungi. Their mechanism of action can be extraordinarily diverse. The joint use of both agents inhibiting the growth of fungi strenghtens the intensity and duration of each other's effect, which may be greater than the algebraical sum of the separate effects; consequently, statins and the antifungal agents show synergic interaction, thus, their joint use can result in the production of a more effective antimycotic composition.
The present invention is illustrated by the following examples. Example 1 : Study of the interactions of combinations comprising fluvastatin and ketoconazole in vitro in Candida albicans
Synergism of fluvastatin and ketoconazole was demonstrated by checkerboard-titration. Checkerboard-titration is a method commonly known for studying effect of substances or combination of substances as well as for demonstrating synergic effect of combinations in vitro. The micro-dilution method used for cultivating yeasts and for determining their sensibility against antibiotics is disclosed in international standard No. NCCLS M27A (NCCLS = National Committee for Clinical Laboratory Standards). The present studies were carried out according to this standards. Cultivation of fungi occured in 96 well microliter plates (Costar 3599). The standard culture medium was RPMI 1640 (Sigma R4130) adjusted to pH=7 with 0.165 M morpholinopropanesulfonic acid (MOPA) and 10 M NaOH. In the case of fluvastatin (Lescol, Novartis) 40 mg of the active agent were dissolved in 1 ml of methanol and further diluted with methanol, and the stock-solution of a concentration of 2,5 mg/ml so obtained was used for producing series of dilution. The stock-solutions of fluvastatin were prepared always freshly. In the case of ketoconazole (Sigma) a stock-solution of a concentration of 5 mg/ml was prepared by dissolving the compound in dimethylsulphoxide (DMSO) which was then diluted with DMSO to a concentration of 1.6 mg/ml. Stock-solutions of ketoconazole were stored at a temperature of -70°C for at most 6 months. Stock-solutions produced from the store of -70°C were used in the studies.
Series of halving dilutions were prepared with the solvents used in the stock-solutions, in the case of fluvastatin by 7 dilution-stages, in the case of ketoconazole by 10 dilution-stages, then each series of 5 dilution was diluted twenty- five-fold with RPMI 1640 medium. Candida albicans (ATCC 90028) cultured in potato-dextrose medium (PDA) incubated for one day at a temperature of 35°C was used in the experiments. The fungus cells were washed off from the agar slants with distilled water, then diluted with RPMI 1640 to the appropriate concentration. Counting of cells was carried out with the aid of a Burker chamber, 1 * 103 cells/well were used as inoculum. 50 μl of twenty-five-fold diluted fluvastatin, 50
10 μl of twenty-five-fold diluted ketoconazole and 100 μl of cell suspension were added to each well of the microtiter plate. Consequently, in the wells of the microtiter plate the concentrations of fluvastatin were 25 through 0.39 μg/ml in the series of halving dilution, the concentrations of ketoconazole were 16 through 0.03 μg/ml in the series of halving dilution.
In this study the following controls were used: growth-control: 100 μl of RPMI 1640 medium and
1 5 100 μl of cell suspension (in RPMI 1640 solution); solvent-control: 100 μl of RPMI 1640 medium containing 1% of solvent and 100 μl of cell suspension (in RPMI 1640 medium); background: 200 μl of RPMI 1640 medium. The samples were cultured at a temperature of 5°C for 3 days and the growth of the cultures was monitored at 620 nm with a microtitre plate reader (Jupiter HD, ASYS Hitech GmbH) after 24, 48 and 72 hours. The obtained data were evaluated with the aid of Microsoft Excel computer-
20 program. The assays were carried out in triplicate. After calculating the avarage and subtracting the background value, the obtained absorbance values were compared with that of growth-control (100%) for calculting the inhibition exerted by fluvastatin and ketoconazole. The interaction (IR) between fluvastatin and ketoconazole was calculated using the above-mentioned Abbott's formula:
!"■ — ^obtained' 'anticipated 25 Iant,c,pa,ed = X + Y - (XY/100) wherein X represents the percentage of inhibition caused by the fluvastatin itself and
Y represents the percentage of inhibition caused by ketoconazole itself.
If the obtained value is 0.5 < IR < 1.5, the interaction is additive; if IR > 1.5, the interaction is synergic, whereas, if IR < 0.5, the interaction is antagonistic. 30 Results
The values of interactions, as calculated with the Abbott's formula, were additive but nearly synergic. Ketoconazole itself causes an inhibition of about 70% even at the lowest concentration, but the level of inhibition cannot be increased by using higher concentrations. Fluvastatin itself causes a total inhibition at a concentration of 6.25 μg/ml, but the use of combinations at concentrations as low as 0.78 35 μg/ml of fluvastatin and 0.06 μg/ml of ketoconazole or 1.56 μg/ml of fluvastatin and 0.03 μg/ml of ketoconazole can result in a total inhibition. The results of Example 1 are summarized in Table 1. Table 1 Interactions of combinations comprising fluvastatin and ketoconazole
Example 2: Study of the interactions of combinations comprising pravastatin and miconazole in vitro in Candida glabrata
Synergism of pravastatin and miconazole was also demonstrated by checkerboard-titration, the method of which was the same as disclosed in Example 1.
In the case of pravastatin (Sigma) 25 mg of the active agent were dissolved in 1 ml of distilled water, then further diluted with distilled water, and the stock-solution of a concentration of 2.5 mg/ml so obtained was used for producing series of dilution. In the case of miconazole (Sigma) a stock-solution of a concentration of 5 mg/ml was prepared by dissolving the compound in DMSO and then diluting with DMSO to a concentration of 1.6 mg/ml. Stock-solutions were stored at a temperature of -70°C for at most 6 months. Stock-solutions produced from the store of -70°C were used in the studies. Series of halving dilutions were prepared with the solvents used in the stock-solutions, in the casse of pravastatin by 7 dilution-stages, in the case of miconazole by 10 dilution-stages, then each series of dilution was diluted twenty-five-fold with RPMI 1640 medium. Candida glabrata (CBS 138) cultured in PDA medium incubated for one day at a temperature of 350C was used in the experiments. The fungus cells were washed off from the agar slants with distilled water, then diluted with RPMI 1640 to the appropriate concentration. Counting of cells was carried out with the aid of a Burker chamber, l * 103 cells/well were used as inoculum. 50 μl of twenty-five-fold diluted pravastatin, 50 μl of twenty-five-fold diluted miconazole and 100 μl of cell suspension were added to each well of the microtiter plate. Consequently, in the wells of the microtiter plate the concentrations of pravastatin were 25 through 0.39 μg/ml in the series of halving dilution, the concentrations of miconazole were 16 through 0.03 μg/ml in the series of halving dilution.
Controls were the same as those in Example 1. The calculation of interaction was carried out with the above-mentioned Abbott's formula. Results
The interactions as calculated with the Abbott's formula are additive in most instances, in the case . of two combinations they are nearly synergic. Pravastatin when used by itself does not show any inhibition of growth, whereas miconazole used by itself causes total inhibition even at a concentration of 2 μg/ml. The concentration of miconazole necessary for a total inhibition can be reduced by one or two dilution stage(s), thus a total inhibition of growth can be achieved by the use of combinations at concentrations as low as 3.125 μg/ml of pravastatin and 0.5 μg/ml of miconazole or 0.78 μg/ml of pravastatin and 1 μg/ml of miconazole. The results of Example 2 are summarized in Table 2.
Table 2
Interactions of combinations comprising pravastatin and miconazole in vitro in Candida slabrata
Example 3:
Study of the interactions of combinations comprising simvastatin and primycin in vitro in Candida glabrata Synergism of simvastatin and primycin was also demonstrated by checkerboard-titration, the method of which was the same as disclosed in Example 1.
In the case of simvastatin (Vasilip, Egis) 40 mg of active agent were dissolved in 1 ml of methanol, then the prodrug form was transformed into open-chain acid form by basic hydrolysisn that is by incubating in a solution containing 15% of ethanol and 0.25% by weight/volume of NaOH at a temperature of 600C for 60 minutes. The solution was further diluted with methanol, and the stock- solution of a concentration of 2.5 mg/ml so obtained was used for producing series of dilution. The stock- solutions were always freshly prepared. In the case of primycin (Pannonpharma) a stock-solution of a concentration of 10 mg/ml was prepared by dissolving the compound in DMSO and then diluting with DMSO to a concentration of 6.4 mg/ml. Stock-solutions were stored at a temperature of -700C for at most 6 months. Stock-solutions produced from the store of -7O0C were used in the studies.
Series of halving dilutions were prepared with the solvents used in the stock-solutions, in the case of simvastatin by 7 dilution-stages, in the case of primycin by 10 dilution-stages, then each series of dilution was diluted twenty-five-fold with RPMI 1640 medium. Candida glabrata (CBS 138) cultured in PDA medium incubated for one day at a temperature of 35°C was used in the experiments. The fungus cells were washed off from the agar slants with distilled water, then diluted with RPMI 1640 to the appropriate concentration. Counting of cells was carried out with the aid of a Burker chamber, l χ lθ3 cells/well were used as inoculum. 50 μl of twenty-five-fold diluted simvastatin, 50 μl of twenty-five-fold diluted primycin and 100 μl of cell suspension were added to each well of the microtiter plate. Consequently, in the wells of the microtiter plate the concentrations of simvastatin were 25 through 0.39 μg/ml in the series of halving dilution, the concentrations of primycin were 16 through 0.03 μg/ml in the series of halving dilution.
Controls were the same as those in Example 1. The calculation of interaction was carried out with Abbott's formula. Results
The interactions as calculated with the Abbott's formula were additive, in the case of 5 combinations they are nearly synergic. Simvastatin by itself shows an inhibition of 50% used at a concentration of 25 μg/ml, whereas primycin by itself causes a total inhibition at a concentration of 32 μg/ml. The concentration of primycin necessary for total inhibition can be reduced by one or two dilution stage(s), thus a total inhibition of growth can be achieved by the use of combinations at concentrations as low as 3.125 μg/ml of simvastatin and 8 μg/ml of primycin or 0.39 μg/ml of simvastatin and 16 μg/ml of primycin. The results of Example 3 are summarized in Table 3.
Table 3
Interactions of combinations comprising simvastatin and primycin in vitro in Candida slabrata
Example 4:
Study of the interactions of combinations comprising atorvastatin and itraconazole in vitro in
Aspergillus fumigatus Synergism of atorvastatin and itraconazole was also demonstrated by checkerboard-titration, the method of which was the same as disclosed in Example 1.
In the case of atorvastatin (Atorvox, Richter) 20 mg of active agent were dissolved in 1 ml of methanol, then the solution was further diluted with methanol, and the stock-solution of atorvastatin of a concentration of 2.5 mg/ml so obtained was used for producing series of dilution. The stock-solutions were always freshly prepared. In the case of itraconazole (Sigma) a stock-solution of a concentration of 5 mg/ml was prepared by dissolving the compound in DMSO and then diluting with DMSO to a concentration of 0.8 mg/ml. Stock-solutions were stored at a temperature of -700C for at most 6 months. Stock-solutions produced from the store of -700C were used in the studies. Series of halving dilutions were prepared with the solvents used in the stock-solutions, in the casse of atorvastatin by 7 dilution-stages, in the case of itraconazole by 10 dilution-stages, then each series of dilution was diluted twenty-five-fold with RPMI 1640 medium. Aspergillus fumigatus cultured in PDA medium incubated for 4 days at a temperature of 350C was used in the experiments. The filamentous fungus spores were washed off from the agar slants with distilled water, then diluted with RPMI 1640 to the appropriate concentration. Counting of cells was carried out with the aid of a Burker chamber, 1 * 104 spores/well were used as inoculum. 50 μl of twenty-five-fold diluted atorvastatin, 50 μl of twenty-fivefold diluted itraconazole and 100 μl of spore susupension were pipetted into each well of the microtiter plate. Consequently, in the wells of the microtiter plate the concentrations. of atorvastatin were 25 through 0.39 μg/ml in the series of halving dilution, the concentrations of itraconazole were 8 through 0.015 μg/mi in the series of halving dilution.
Controls were the same as those in Example 1. The calculation of interaction was carried out with Abbott's formula. Results
The interactions as calculated with the Abbott's formula are synergic in most instances. Atorvastatin by itself shows an inhibition of 40% used at a concentration of 25 μg/ml, whereas itraconazole by itself causes a total inhibition at a concentration of 0.25 μg/ml. The concentration of itraconazole necessary for a total inhibition can be reduced by one or two dilution stage(s), thus a total inhibition of growth can be achieved by the use of combinations at concentrations as low as 6.25 μg/ml of atorvastatin and 0.06 μg/ml of itraconazole or 0.39 μg/ml of atorvastatin and 0.125 μg/ml of itraconazole. The results of Example 4 are summarized in Table 4.
Table 4
Interactions of combinations comprising atorvastatin and itraconazole in vitro in Aspergillus fumigatus
Example 5:
Study of the interactions of combinations comprising fluvastatin and miconazole in vitro in
Aspergillus flavus
Synergism of fluvastatin and miconazole was also demonstrated by checkerboard-titration, the method of which was the same as disclosed in Example 1.
In the case of fluvastatin (Lescol, Novartis) 40 mg of active agent were dissolved in 1 ml of methanol, then the solution was further diluted with methanol, and the stock-solution of a concentration of 2.5 mg/ml so obtained was used for producing series of dilution. The stock-solutions were always freshly prepared. In the case of miconazole (Sigma) a stock-solution of a concentration of 5 mg/ml was prepared by dissolving the compound in DMSO and then diluting with DMSO to a concentration of 1.6 mg/ml. Stock-solutions were stored at a temperature of -700C for at most 6 months. Stock-solutions produced from the store of -70°C were used in the studies.
Series of halving dilutions were prepared with the solvents used in the stock-solutions, in the casse of fluvastatin by 7 dilution-stages, in the case of miconazole by 10 dilution-stages, then each series of dilution was diluted twenty-five-fold with RPMI 1640 medium. Aspergillus flavus cultured in PDA medium incubated for 4 days at a temperature of 35°C was used in the experiments. The filamentous fungus spores were washed off from the agar slants with distilled water, then diluted with RPMI 1640 to the appropriate concentration. Counting of cells was carried out with the aid of a Burker chamber, 1 * 104 spores/well were used as inoculum. 50 μl of twenty-five-fold diluted fluvastatin, 50 μl of twenty-five-fold diluted miconazole and 100 μl of spore suspension were pipetted into each well of the microtiter plate. Consequently, in the wells of the microtiter plate the concentrations of fluvastatin were 25 through 0.39 μg/ml in the series of halving dilution, the concentrations of miconazole were 16 through 0.03 μg/ml in the series of halving dilution.
Controls were the same as those in Example 1. The calculation of interaction was carried out with Abbott's formula. Results
The interactions as calculated with the Abbott's formula are additive in most instances, but there are a number of synergic combinations, too. Fluvastatin by itself does not cause a significant inhibition of growth even at the highest concentration used, whereas miconazole by itself causes a total inhibition at a concentration of 4 μg/ml. The concentration of miconazole necessary for total inhibition can be reduced by one or two dilution stage(s) when combined with fluvastatin, thus a total inhibition of growth can be achieved by the use of combinations at concentrations as low as 6.25 μg/ml of fluvastatin and 1 μg/ml of miconazole or 0.78 μg/ml of fluvastatin and 2 μg/ml of miconazole. The results of Example 5 are summarized in Table 5. Table 5
Interactions of combinations comprising fluvastatin and miconazole in vitro in Aspergillus flavus
Example 6: Study of the interactions of combinations comprising atorvastatin and ketoconazole in vitro in Rhizopus oryzae
Synergism of atorvastatin and ketoconazole was also demonstrated by checkerboard-titration, the method of which was the same as disclosed in Example 1.
In the case of atorvastatin (Atorvox, Richter) 20 mg of active agent were dissolved in 1 ml of methanol, then the solution was further diluted with methanol, and the stock-solution of a concentration of 2.5 mg/ml so obtained was used for producing series of dilution. The stock-solutions were always freshly prepared. In the case of ketoconazole (Sigma) a stock-solution at a concentration of 5 mg/ml was prepared by dissolving the compound in DMSO and then diluting with DMSO to a concentration of 1.6 mg/ml. Stock-solutions were stored at a temperature of -7O0C for at most 6 months. Stock-solutions produced from the store of -700C were used in the studies.
Series of halving dilutions were prepared with the solvents used in the stock-solutions, in the casse of atorvastatin by 7 dilution-stages, in the case of ketoconazole by 10 dilution-stages, then each series of dilution was diluted twenty-five-fold with RPMI 1640 medium. Rhizopus oryzae (CBS 109.939) cultured in PDA medium incubated for 4 days at a temperature of 350C was used in the experiments. The filamentous fungus spores were washed off from the agar slants with distilled water, then diluted with RPMI 1640 to the appropriate concentration. Counting of cells was carried out with the aid of a Burker chamber, l * 104 spores/well were used as inoculum. 50 μl of twenty-five-fold diluted atorvastatin, 50 μl of twenty- five-fold diluted ketoconazole and 100 μl of spore suspension were pipetted into each well of the microtiter plate. Consequently, in the wells of the microtiter plate the concentrations of atorvastatin were 25 through 0.39 μg/ml in the series of halving dilution, the concentrations of ketoconazole were 16 through 0.03 μg/ml in the series of halving dilution.
Controls were the same as those in Example 1. The calculation of interaction was carried out with Abbott's formula. Results
The interactions as calculated with the Abbott's formula are synergic in most instances. Atorvastatin by itself shows an inhibition of 60% used at a concentration of 25 μg/ml, whereas ketoconazole by itself causes a total inhibition at a concentration of 4 μg/ml. The concentration of ketoconazole necessary for a total inhibition can be reduced by one or two dilution stage(s) when used in combination with atorvastatin, thus a total inhibition of growth can be achieved by the use of combinations at concentrations as low as 3.125 μg/ml of atorvastatin and 1 μg/ml of ketoconazole or 0.39 μg/ml of atorvastatin and 2 μg/ml of ketoconazole. The results of Example 6 are summarized in Table 6.
Table 6
Interactions of combinations comprising atorvastatin and ketoconazole in vitro in Rhizopus orvzae
If the statin used in combination was different from simvastatin, fluvastatin, atorvastatin and pravastatin, the stock-solutions were prepared in the following way. In the case of lovastatin (Mevacor, MSD) 40 mg of the active agent were dissolved in 1 ml of methanol, then the prodrug form was transformed into active open-chain acid form by basic hydrolysis, that is by carrying out incubation in a solution containing 15% of ethanol and 0.25% by weight/volume of NaOH at a temperature of 6O0C for 60 minutes. The solution was further diluted with methanol, and the stock-solution of lovastatin of a concentration of 2.5 mg/ml so obtained was used for producing series of dilution. In the case of rosuvastatin 10 mg of the active agent were dissolved in 1 ml of methanol, then the solution was further diluted with methanol, and the stock-solution of rosuvastatin of a concentration of 2.5 mg/ml so obtained was used for producing series of dilution. The stock-solutions were always freshly prepared.
If the used antifungal agent was different from ketoconazole, miconazole, itraconazole and primycin, the stock-solutions were prepared in the following way. In the case of fluconazole or griseofulvin a stock-solution of a concentration of 10 mg/ml was prepared by dissolving the compound in dimethylformamid (DMF) and then diluting with DMF to a concentration of 6.4 mg/ml. In the case of nystatin a stock-solution of a concentration of 5 mg/ml (22,000 U) was prepared by dissolving the active compound in dimethylsulphoxide, which was then diluted with DMSO to a concentration of 0.8 mg/ml. In the case of terbinafine a stock-solution of a concentration of 5 mg/ml was prepared by dissolving the active compound in dimethylsulphoxide and then diluting with DMSO to a concentration of 0.8 mg/ml. Stock-solutions were stored at a temperature of -7O0C for at most 6 months. Stock-solutions produced from the store of -7O0C were used in the studies. The concentration of the factory-made stock-solution of amphotericin B is 250 μg/ml. It was stored at a temperature of -2O0C in a package protecting from light. In the studies carried out in connection with the present invention synergic interactions between the statins and antifungal agents have been observed in the case of a number of combinations. The level of interactions have been determined with the aid of Abbott's formula. This calculation formula takes into consideration the values of the measured absorbance and the inhibition values claculated therefrom, whereas in the case of other formulas the calculation is based on the subjectively determined value of MIC (Minimal Inhibitory Concentration). In many cases, however, the calculation with the aid of Abbott's formula demonstrates that the interaction is not synergic but only additive, nevertheless by the use of both agents together a higher level of inhibition can be achieved than those by themselves, or else, a total inhibition can be achieved at lower concentration if the agents are used in combination. In a number of cases it can be observed among the combinations of statins and antifungal agents that the combined use results in a higher effect. However, the inhbition of growth occurs not always at a clinically reachable concentration. The statin/antifungal agent combinations are stressed in Table 7 which have shown high levels of inhibition of growth in the case of most fungi even at low concentrations and which show extraordinary effectiveness in the case of certain groups of fungi.
Table 7 Statin/antimvcotic agent combinations showing high levels of inhibition of growth in the case of most fungi even at low concentrations and combinations, respectively, showing extraordinary effectiveness in the case of certain groups of fungi
γ™ Shows high levels of inhibition of growth in the case of all studied fungi even at low concentrations
XX Shows high levels of inhibition of growth in the case of most of the studied fungi even at low concentrations
X Combinations showing effectiveness in the case of certain groups of fungi 0 Synergic interaction cannot be observed

Claims

Claims
1. A combination comprising (a) one or more active component(s) having antimycotic activity selected from the group consisting of amphotericin B, nystatin, griseofulvin, primycin, terbinafme, compounds of azole type, preferably, ketoconazole, miconazole, fluconazole, itraconazole, clotrimazole, posaconazole, voriconazole, and prodrugs and metabolites thereof, and (b) one or more statin(s), selected from the group consisting of lovastatin, simvastatin, fluvastatin, pravastatin, rosuvastatin, atorvastatin, cerivastatin, pitavastatin, prodrug and metabolites thereof for the use as an agent inhibiting the growth of fungi, with the proviso that the combination is different from amphotericin B and fluvastatin; ketoconazole and lovastatin; fluconazole and lovastatin; fluconazole and fluvastatin; fluconazole and pravastatin; itraconazole and fluvastatin; voriconazole and lovastatin; and voriconazole and fluvastatin.
2. A combination as claimed in claim 1 comprising one or more active component(s) having antimycotic activity and one or more statin(s) or prodrugs or metabolites of the said compounds for the use as an agent inhibiting the growth of fungi adminisered in clinically acceptable dose.
3. A combination as claimed in claim 1 or 2 comprising an active component having antimycotic activity, prodrugs or metabolites thereof in an amount of
(i) in the case of amphotericin B if administered intravenously, 0.01 to 2 mg/kg body weight/day, if administered orally, 10 to 10,000 mg/day, if adminstered topically, a concentration of 0.001 to 5 % by weight;
(ii) in the case of nystatin if administered orally or topically, an amount of 10,000 to 10,000,000 USP Nystatin-Unit;
(iii) in the case of grisoufulvin if administered orally, 0.1 to 2000 mg/day, preferably 0.1 to 1000 mg/day, more preferably 0.1 to 250 mg/day;
(iv) in the case of primycin if administered topically, a concentration of 0.02 to 5 % by weight; (v) in the case of terbinafme if administered orally, 0.1 to 400 mg/day, preferably 0.1 to 100 mg/day, more preferably 0.1 to 50 mg/day; if administered topically, a concentration of 0.001 to 5 % by weight; (vi) in the case of an antimycotic agent of azole type if administered orally, 0.1 to 1200 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if administered topically, a concentration of 0.001 to 5 % by weight; (vii) in the case of ketoconazole if administered orally, 0.1 to 1200 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if administered topically, a concentration of 0.001 to 5 % by weight; (viii) in the case of miconazole if administered orally, 0.1 to 1000 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if administered topically, a concentration of 0.001 to 5 % by weight;
(ix) in the case of fluconazole if administered orally, 0.1 to 1200 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if administered topically, a concentration of 0.001 to 5 % by weight; (x) in the case of itraconazole if administered orally, 0.1 to 1200 mg/day, preferably 0,1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if administered intravenously, 0.01 to 20 mg/kg body weight/day; if adminstered topically, a concentration of 0.001 to 5 % by weight;
(xi) in the case of clotrimazole if administered orally, 0.1 to 500 mg/day, preferably 0.1 to 250 mg/day, more preferably 0.1 to 100 mg/day; if administered topically, a concentration of 0.001 to 5 % by weight; (xii) in the case of posaconazole if administered orally, 0.1 to 800 mg/day, preferably 0.1 to 400 mg/day, more preferably 0.1 to 100 mg/day; if administered topically, a concentration of 0.001 to 5 % by weight; (xiii) in the case of voriconazole if administered orally, 0.1 to 400 mg/day, preferably 0.1 to 100 mg/day, more preferably 0.1 to 50 mg/day; if administered intavenously, 0.1 to 12 mg/kg body weight/day; if administered topically, a concentration of 0.001 to 5 % by weight; (xiv) in the case of lovastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day; (xv) in the case of simvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day; (xvi) in the case of fluvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 20 mg/day;
(xvii) in the case of pravastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day;
(xviii) in the case of rosuvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 5 mg/day; (xix) in the case of atorvastatin 0.1 to 160 mg/day, preferably 0.1 to 80 mg/day, more preferably 0.1 to 10 mg/day; (xx) in the case of cerivastatin 0.025 to 5 mg/day, preferably 0.025 to 2,5 mg/day, more preferably 0.025 to 1 mg/day; (xxi) in the case of pitavastatin 0.1 to 80 mg/day, preferably 0.1 to 40 mg/day, more preferably 0.1 to 5 mg/day for the use as an agent inhibiting the growth of fungi.
4. A combination as claimed in any of claims 1 to 3 for the use as an agent inhibiting the growth of fungi selected from the group of Candida spp. (such as Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida guillennondii, Candida knisei and Candida kefyr), Cryptococciis spp. (such as Ciγptococciis neoformans), Pneumocystis carinii, Aspergillus spp. (such as Aspergillus flavus, Aspergillus fiimigatus, Aspergillus terreits, Aspergillus repens, Aspergillus versicolor), Mucor spp. (such as Mucor circinelloides, Mucor indicus, Mucor ramosissimus), Rhi∑oimtcor spp. (such as Rhizomucor piisillus, Rhizomucor miehei), Rhizopυs spp. (such as Rhizopus ory∑ae, Rhizopus microsporia var. microporus, Rhizopus microsporus var. rhi∑opodiformis, Rhizopus schipperae), Absidia (such as Absidia corymbifera), Apophysomyces (such as Apophysoinyces elegans), Cunninghamella (such as Cimninghamella bertholletiae), Saksenaea (such as Saksenaea vasiformis), Cokeromyces (such as Cokeromyces recurvatiis), Syncephalastnim (such as Syncephalastnim racemoswn), Basidiobolus (such as Basidiobolus rananim), Conidiobolus (such as Conidiobohis coronatus), Coccidioides spp. (such as Coccidioides immitis), Paracoccidioides spp. (such as Paracoccidioides brasiliemis), Histoplasma spp. (such as Histoplasma capsulatum) or Blastomyces spp. (such as Blastomyces dennatitidis), Trichophyton spp. (such as Trichophyton mentagrophytes, Trichophyton rubnim), Microsporum spp. (such as Microsporum canis, Microsporum gypseum) or Epidermophyton spp. (such as (Epidermophyton floccosum) and Pityrosporum spp. [such as Pityrosporum orbiculare (MaI ass e∑i a furfur)], Geotrichum (such as Geotrichum clavatum, Geotrichum candidum), Trichosporon (such as Trichosporon beigelii), Blastoschizomyces (such as Blastoschizomyces capitatus), Sporothrix (such as Sporothrix schenckiϊ), Scedosporium (such as Scedosporiiim apiosperum, Scedosporium prolificans), Cladosporium (such as Cladosporium cladosporioides, Cladosporium sphaerospermum), Acremonhim spp. (such as Acremonium falciforme), Beauveria spp. (such as Beauveria bassiana), Fusarium spp. (such as Fusarium oxysporum, Fusarium solani, Fusarium moniliforme), Paecilomyces spp. (such as Paecilomyces variotii, Paecilomyces lilacinus, Paecilomyces marquandii), Penicillium spp. (such as Penicillium marneffei, Penicillium purpurogenum), Scopulariopsis spp. (such as Scopulariopsis brevicaulis), Cladophialophora spp. (such as Cladophialophora bantiana, Cladophialophora carrionii), Aureobasidium spp. (such as Aureobasidium pullulans), Bipolaris spp. (such as Bipolaris australiensis, Bipolaris hawaiiensis, Bipolaris spicifera), Curvularia spp. (such as Curvularia lunata, Curvularia pallescens, Curvularia geniculata), Exophiala spp. (such as Exophiala spinifera), Exserohilum (such as Exserohilum rostratum), Phialophora spp. (such as Phialophora parasitica), Wangiella spp. (such as Wangiella dennatitidis), Alternaria spp. (such as Alternaria alternata), Chaetomium spp. (such as Chaetomium globosum), Coniophora spp. (such as Coniophora puteana), preferably Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida guillermondii, Candida krusei, Candida kefyr, Aspergillus flavus, Aspergillus fumigatus, Rhi∑opus oiyzae, Trichophyton mentagrophytes, Trichophyton rubnim, Microsporum canis, Microsporum gypseum, Malasse∑ia pachydermatis.
5. A combination composition having antimycotic effect comprising a clinically acceptable dose of a combination of an active component having antimycotic activity and a statin, a prodrug or a metabolite of the said compounds selected from the goup consisting of (i) amphotericin B and rosuvastatin, (ii) amphotericin B and pitavastatin, (iii) primycin and rosuvastatin, (iv) primycin and cerivastatin, (v) primycin and pitavastatin, (vi) voriconazole and simvastatin, (vii) voriconazole and pravastatin, (viii) voriconazole and atorvastatin, and (ix) voriconazole and cerivastatin, and optionally one or more pharmaceutically acceptable excipients.
6. A combination composition as claimed in claim 5 containing the active components in the following amounts: in the case of combination (i), 0.1 to 99 % by weight of amphotericin B and 0.1 to 99 % by weight of rosuvastatin; in the case of combination (ii), 0.1 to 99 % by weight of amphotericin B and 0.1 to 99 % by weight of pravastatin, in the case of combination (iii), 0.1 to 99 % by weight of primycin and 0.1 to 99 % by weight of rosuvastatin, in the case of combination (iv), 0.1 to 99 % by weight of primycin and 0.1 to 99 % by weight of cerivastatin, in the case of combination (v), 0.1 to 99 % by weight of primycin and 0.1 to 99 % by weight of pitavastatin, in the case of combination (vi), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of simvastatin, in the case of combination (vii), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of pravastatin, in the case of combination (viii), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of atorvastatin, in the case of combination (ix), 0.1 to 99 % by weight of voriconazole and 0.1 to 99 % by weight of cerivastatin.
7. A combination composition as claimed in claim 5 or 6 characterized by being effective against the following fungi: Candida spp. (such as Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida guillermondii, Candida knisei and Candida kefyr), Ciγptococcus spp. (such as Cryptococcus neoformans), Pneumocystis carinii, Aspergillus spp. (such as Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Aspergillus repens, Aspergillus versicolor), Mucor spp. (such as Mucor circinelloides, Mucor indicus, Mucor ramosissimus), Rhizomucor spp. (such as Rhizomucor pusillus, Rhizomucor miehei), Rhizopus spp. (such as Rhizopus oryzae, Rhizopus microsporus var. microporus, Rhizopus microsporus var. rhizopodiformis, Rhizopus schipperae), Absidia (such as Absidia corymbifera), Apophysomyces (such as Apophysomyces elegans), Cunninghamella (such as Cunninghamella bertholletiae), Saksenaea (such as Saksenaea vasiformis), Cokeromyces (such as Cokeromyces recurvatus), Syncephalastrum (such as Syncephalastrum racemosum), Basidiobolus (such as Basidiobolus ranarum), Conidiobolus (such as Conidiobolus coronatus), Coccidioides spp. (such as Coccidioides immitis), Paracoccidioides spp. (such as Paracoccidioides brasiliensis), Histoplasma spp. (such as Histoplasma capsulatum) or Blastomyces spp. (such as Blastomyces dermatitidis), Trichophyton spp. (such as Trichophyton mentagrophytes, Trichophyton riibntm), Microsporum spp. (such as Microsporum canis, Microsporum gypseum) vagy Epidermophyton spp. (such as (Epidermophyton floccosum) and Pityrosporum spp. [such as Pityrosporum orbiculare (Malassezia furfur)], Geotήchum (such as Geotrichum clavatum, Geotrichum candidum), Trichosporon (such as Trichosporon beigelii), Blastoschizomyces (such as Blastoschizomyces capitatus), Sporothrix (such as Sporothrix schenckii), Scedosporium (such as Scedosporium apiosperum, Scedosporium prolificans), Cladosporium (such as Cladosporium cladosporioides, Cladosporium sphaerospermum), Acremonium spp. (such as. Acremonium falciforme), Beauveria spp. (such as Beauveria bassiana), Fusarium spp. (such as Fusarium oxysponim, Fusarium solani, Fusarium moniliforme), Paecilomyces spp. (such as Paecilomyces variotii, Paecilomyces lilacinus, Paecilomyces marquandii), Penicillium spp. (such as Penicillium mameffei, Penicillhim purpurogenum), Scopiilariopsis spp. (such as Scopiilariopsis brevicaiilis), Cladophialophora spp. (such as Cladophialophora bantiana, Cladophialophora carrionii), Aureobasidium spp. (such as Aureobasidium pullulans), Bipolaris spp. (such as Bipolaris austr aliens is, Bipolaris hawaiiemis, Bipolaris spicifera), Ciirvidaria spp. (such as Ciirvularia hmata, Curvularia pallescens, Curvularia geniciύatά) , Exophiala spp. (such as Exophiala spinifera), Exserohilum (such as Exserohilum rostratum), Phialophora spp. (such as Phialophora parasitica), Wangiella spp. (such as Wangiella dermatitidis), Alternaria spp. (such as Alternaria alternata), Chaelomium spp. (such as Chaetomium globosiim), Coniophora spp. (such as Coniophora puteanά), preferably Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida guillermondii, Candida krusei, Candida kefyr, Aspergillus flaviis, Aspergillus fumigatus, Rhizopus oryzae, Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis, Microsporum gypseum, Pityrosporum orbiculare.
8. A combination composition as claimed in any of claims 5 to7 characterized by being formulated as a medicine, preferably as a medicine for internal or external administration, particularly in the form of tablet, capsule, cream, gel, ointment, powder, lacquer, solution, or foam.
9. A combination composition as claimed in any of claims 5 to7 for the use as medicine, preferably as a medicine having antimycotic effect.
10. Use of a combination of active components as claimed in claim 5 for preparing a composition having enhanced antimycotic effect.
EP09742410A 2008-05-09 2009-05-07 Combination compositions comprising antimycotic agents and statins and the use thereof Withdrawn EP2296644A1 (en)

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