| PATENT NUMBER | This data is not available for free |
| PATENT GRANT DATE | April 24, 1990 |
| PATENT TITLE |
Antifungal compositions and method of controlling mycotic infections |
| PATENT ABSTRACT | Method of potentiating antifungal agents with 25-azasterol compound to produce superior antifungal compositions and use of the compositions to control mycotic infections is disclosed |
| PATENT INVENTORS | This data is not available for free |
| PATENT ASSIGNEE | This data is not available for free |
| PATENT FILE DATE | April 18, 1988 |
| PATENT PARENT CASE TEXT |
Chemical Abstracts; vol. 85 (1976); #187178k; Kabara et al. Chemical Abstracts; vol. 94 (1981); #101307b; Monaghan et al. J. Chem. Soc. (C), 1971, (23), pp. 3888-3891; Aldridge et al. J. Chem. Soc. (D), 1970 (11), p. 639; Aldridge et al. Ansehn, S et al., Scand. J. Infect. Dis., Suppl. 9, 62-66 ('76), Lew M. A. et al., J. Infect. Dis. 136, 263-270 (1977). Medoff, G., et al., Proc. Nat. Acad. Sci. 69, 196-199 (1972). Oehlschlager, A. C., et al., Biochem. 23, 3582-3589 (1984). Gordee, R. S., et al., J. Antibiotics, 28, 112 (1975). Rahier, A., et al., J. Biol. Chem. 259, No. 24, 15215 (1984). |
| PATENT CLAIMS |
What is claimed is: 1. A method for potentiating the antifungal properties of an antifunga agent comprising coadministering a 25-azasterol compound with a non-steroidal antifungal agent. 2. A method according to claim 1 wherein the 25-azasterol compound is a compound represented by the formula ##STR19## wherein R is hydrogen or lower acyl, X is CH, CH.sub.2 or O; and the on the side chain indicates that the bond may be a single or a double bond provided that when X is O, or CH.sub.2, it is a single bond. 3. A method according to claim 1 wherein the 25-azasterol compound is 17.beta.-[[3-(dimethylamino)-propyl]methylamino]androst-en-3.beta.-ol (25-azacholesterol). 4. A method according to claim 1 wherein the non-steroidal antifungal agent is employed in a subfungistatic amount and the 25-azasterol compound is employed in an amount not inhibitory to fungal growth. 5. A method according to claim 1 in which the antifungal compound is a lanosterol synthesis inhibitor. 6. A method according to claim 5 in which the antifungal compound is a mevalonic acid synthesis inhibitor. 7. A method according to claim 6 in which the mevalonic acid synthesis inhibitor is a HMG-CoA synthase inhibitor. 8. A method according to claim 7 in which the HMG-CoA synthase inhibitor is 11-(3-(hydroxymethyl)-4-oxo-2-oxethanyl)-7-methyl-2,4-undecadienoic acid. 9. A method according to claim 7 in which the mevalonic acid synthesis inhibitor is a .beta.-ketothiolase inhibitor. 10. A method according to claim 9 in which the .beta.-ketothiolase inhibitor is 5-(1-hydroxy-2,4,6-heptatriynyl)-2-oxo-1,3-dioxolane-4-heptanoic acid. 11. A method according to claim 7 in which the mevalonic acid synthesis inhibitor is a HMG-CoA reductase inhibitor. 12. A method according to claim 11 in which the HMG-CoA reductase inhibitor is 1,2,3,7,8,8a-hexahydro-3,7-dimethyl-8-[2-(tetrahydro-4-hydroxy-6-oxo-2H-py ran-2-yl)ethyl]-1-naphthalenyl-2-methylbutanoate (lovastatin). 13. A method for treating mycotic infections comprising administering subjects with an antifungally effective amount of a composition comprising a subfungistatic amount of a non-steroidal antifungal compound and a 25-azasterol compound in an amount not inhibitory to fungal growth. 14. A method according to claim 13 wherein the 25-azasterol compound is 25-azacholesterol. 15. A method according to claim 13 wherein the non-steroidal antifungal compound is administered in a daily dose in an amount of from about 1.5 to 5.0 mg/kg of body weight and the 25-azasterol compound is employed in an amount which is from about 1.5 to 5.0 mg/kg of body weight. 16. An improved antifungal composition comprising: (1) an azasterol compound represented by the formula ##STR20## wherein r is hydrogen or lower acyl, X is CH, CH.sub.2 or O; and the on the side chain indicates that the bond may be a single or a double bond provided that when X is O, or CH.sub.2, it is a single bond; and (2) a non-steroidal antifungal compound in admixture with a pharmaceutically acceptable carrier. 17. A composition according to claim 16 wherein the azasterol compound is employed in an amount of from about 50 to 200 milligrams and the non-steroidal antifungal compound is employed in an amount of from about 100 to 400 milligrams. 18. A composition according to claim 17 wherein the 25-azasterol compound is 25-azacholesterol. 19. A composition according to claim 17 wherein the 25-azasterol compound is (3.beta.,22Z)-24(dimethylamino)chola-5,22-dien-3-ol. 20. A composition according to claim 17 wherein the 25-azasterol compound is 25-aza-22-oxacholesterol |
| PATENT DESCRIPTION |
BACKGROUND OF THE INVENTION Systemic fungal infections caused by Candida species, Cryptococcus neoformans, Histoplasma capsulatum and the like are often serious or fatal. There continues to be a need for an antifungal agent which is effective and at the same time non-toxic to the patient being treated. A number of known antifungal agents while effective in eliminating the disease are of limited usefulness because of toxic or other undesirable side reactions. In many cases the toxicity is related to the amount of the drug and could be eliminated or reduced if a lesser amount of the drug could be employed. Thus, it would be desirable if a combination could be found which is synergistic or which has a second component that has a potentiating effect on the known antifungal agent thereby reducing the amount of drug required with concomitant reduction or elimination of side reactions. There are reports in the literature that by administering amphotericin with other antibiotics, Candida albicans is sensitized to antifungal agents. Thus, for example, minocycline and amphotericin B have been found to be a synergistic composition against Candida albicans, Leev et al, Brit. J. Infect. Dis., 136, 263-270 (1977). However, combinations with amphotericin is not satisfactory because of the toxicity of amphotericin to human beings. STATEMENT OF THE INVENTION The present invention concerns improved compositions and methods useful for the control of fungi and particularly for the treatment of mycotic infections made possible by the discovery that when certain azasterol compounds are employed in combination with certain fungistatic compounds, there is potentiation in the activity of these compounds resulting in superior antifungal combinations and in some cases resulting in useful fungicidal combinations. Some azasterols may show some antifungal properties in which case the potentiated compositions may be considered a synergistic composition. DESCRIPTION OF THE INVENTION According to the present invention it has been discovered that the antifungal properties of an antifungal agent may be potentiated by coadministering a 25-azasterol compound with the antifungal agent. Thus, when a 25-azasterol compound, at a concentration not inhibitory to the growth of fungi is employed with a subfungistatic amount of an antifungal agent, the sensitivity of fungal organisms to the antifungal agents is unexpectedly significantly increased providing a synergistic antifungal composition. It has further been discovered that with certain fungistats the combinations have resulted unexpectedly in fungicidal compositions. In view of the effectiveness of the combination against fungi causing mycotic infections, the present invention is further directed to methods and compositions for combatting fungi causing mycotic infections comprising treating the infected site with an antifungally effective amount of a composition comprising a subfungistatic amount of an antifungal compound and a 25-azasterol compound in an amount not inhibitory to fungal growth. The invention is further directed to a method for treating mycotic infections comprising administering to a subject in need of such treatment, an antifungally effective amount of a composition comprising a subfungistatic amount of an antifungal compound and an amount of 25-azasterol not inhibitory to fungal growth. Fungistats which have been potentiated most effectively and indeed unexpectedly to give rise to fungicidal compositions are those fungistats which act on the enzymes in the biochemical pathway prior to the formation of lanosterol. Inhibitors of enzymes in the mevalonic acid synthesis such as HMG-CoA synthase, .beta.-ketothiolase (.beta.-ketoacyl-coenzyme A thiolase) and HMG-CoA reductase as well as those which inhibit enzymes which act at later stages in the pathway to lanosterol including squalane epoxidase are particularly important in the formation of novel fungicidal compositions. Since all of these enzymes are enzymes in the pathway of lanosterol synthesis, the antifungal agents which act on these enzymes when spoken of in general terms will be referred to as lanosterol synthesis inhibitors. In view of the effectiveness of 25-azasterols on potentiating the fungistats which are lanosterol synthesis inhibitors, the invention is further directed to methods and compositions for treating mycotic infections comprising a subfungistatic amount of a lanosterol synthesis inhibitor and a 25-azasterol compound in an amount non-inhibitory to fungal growth. Although mycotic infections may be alleviated by fungistatic compositions, there is no cure except with fungicidal compositions. A very special aspect of the present invention are new fungicidal compositions comprising a 25-azasterol compound and a HMG-CoA synthase inhibitor, or a .beta.-ketothiolase inhibitor, or a HMG-CoA reductase inhibitor or a squalene epoxidase inhibitor. The "25-azasterol compound" useful in the improved antifungal compositions and methods of the present invention are characterized by a hydroxyl group or an esterified hydroxyl group in the 3-position, an unsaturation in Ring B and a nitrogen at the 25 position in the side chain of the steroid ring system as may be seen from the following formula (I): ##STR1## wherein R is hydrogen or lower acyl, X is CH, CH.sub.2 or O; and the on the side chain indicates that the bond may be a single or a double bond provided that when X is O, or CH.sub.2, it is a single bond. Except as hereinafter indicated, the azasterol compounds are compounds available or reported in the literature or are ester derivatives of known compounds readily prepared by conventional esterification methods. The following are representative examples of specific 25-azasterols which are particularly useful in the present invention: (a) 25-azacholesterol; 17.beta.-[[3-(dimethylamino)-propyl]methylamino]androst-5-en-3.beta.-ol ##STR2## (b) N,N-dimethyl-3.beta.-hydroxy-5,22(Z)choladiene-24-amine; (3.beta.-22Z)-24-(dimethylamino)chola-5,22-diene-3-ol ##STR3## The foregoing may be prepared by the Wittig reaction on an i-steroid carboxaldehyde, followed by rearrangement and hydrolysis as described in copending application U.S. Ser. No. 169,669 in the name of N. G. Steinberg, now abandoned. (c) N,N-dimethyl-3.beta.-acetoxy-5,22(Z)choladiene-24-amine; (3.beta.,22Z)-24-(dimethylamino)chola-5,22-dien-3-ol acetate. ##STR4## The preparation of the acetate is also the subject of U.S. Ser. No. 169,669 in the name of N. G. Steinberg, now abandoned. (d) 25-aza-22-oxacholesterol; (3,20)-20-[2-(dimethylamino)ethoxy]pregn-5-en-3-ol ##STR5## The preferred 25-azasterol compound is 25-azacholesterol (Compound Ia). The antifungal compounds which may be potentiated by the 25-azasterol compounds are diverse in structure but are useful especially against organisms causing mycotic infections. A number of the suitable antifungal compounds are those known to be inhibitors of enzymes in the biochemical pathway to lanosterol. However, a number of antifungal agents which are not inhibitors of enzymes in the biochemical pathway to lanosterol or where mode of antifungal action is not established but which are known or can be shown to possess antifungal properties have also been found to be potentiated and to form useful synergistic antifungal compositions. These compounds of diverse structure which are the main antifungal component in the methods and compositions of the present invention are non-steroidal in nature, although as previously noted, some of since the 25-azasterol compounds have some antifungal properties. Representative of the compounds which are potentiated by the 25-azasterol compound include a number of new fungistats as well as established fungistats recognizable by USAN or generic names. The following representative compounds illustrate the diverse nature of the antifungal agents which may be potentiated but are not to be construed as being limited thereto. (1) 11-(3-hydroxymethyl-4-oxo-2-oxetanyl)-7-methyl-2,4-undecadienoic acid. (Compound A) ##STR6## This compound is reported in J. Chem. Soc. (c), 1971, 3888; the antifungal properties is the subject of U.S. Ser. No. 825,496, filed Feb. 3, 1986, now abandoned and the HMG-CoA synthase inhibitor property, the subject of U.S. Ser. No. 21,848, filed Mar. 4, 1987, now U.S. Pat. No. 4,847,271, Jul. 11, 1989. (2) 5-(1-hydroxy-2,4,6-heptatriynyl)-2-oxo-1,3-dioxolane-4-heptanoic acid. (Compound B) ##STR7## The compound may be prepared by the cultivation of microorganism ATCC 53,614, 53,615 or 53,616 followed by isolation as described together with antifungal properties in U.S. Ser. No. 53,920, filed May 26, 1987, now U.S. Pat. No. 4,806,565, Feb. 21, 1989. The .beta.-ketothiolase activity is subject of U.S. Ser. No. 53,973, filed May 26, 1987, now U.S. Pat. No. 4,780,311. (3) 1-methyl-2-nonyl-5-(phenylmethyl)-3-pyrrolidinol (Compound C) ##STR8## The compound may be prepared by the cultivation of microorganism ATCC 22947 followed by isolation as described together with antifungal properties in copending application U.S. Ser. No. 172,164, filed in the name of R. E. Schwartz et al., now U.S. Pat. No. 4,847,284, Jul. 11, 1989. (4) (E)-N-(6,6-dimethyl-2-hepten-4-ynyl)-N-methyl-1-naphthalenemethylamine; terbinafine (Compound D) ##STR9## (5) 1,2,3,7,8,8a-hexahydro-3,7-dimethyl-8-[2-(tetrahydro-4-hydroxy-6-oxo-2H-py ran-2-yl)ethyl]-1-naphthalenyl-2-methyl butanoate; lovastatin (Compound E) ##STR10## (6) 1-ethyl-1,4-dihydro-7-methyl-4-oxo-1,8-naphthyridine-3-carboxylic acid; nalidixic acid (Compound F) ##STR11## (7) 5-chloro-6-(7,8-epoxy-10-hydroxy-2-oxo-3,5-undecadienyl)-.beta.-resorcycli c acid .mu.-lactone; monorden (Compound G) ##STR12## (8) 2-(4-thiazolyl)-1H-benzimidazole; thiabendazole (Compound H) ##STR13## (9) 4-[2-(3,5-dimethyl-2-oxocyclohexyl)-2-hydroxyethyl]2,6-piperidinedione; cy cloheximide (Compound J) ##STR14## (10) 4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-actahydro-3,10,12,12a-tetrahyd roxy-1-dioxo-2-naphthacenecarboxamide; minocycline (Compound K) ##STR15## (11) 4-(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,6,10,12,12a-pentahydrox y-6-methyl-1,11-dioxo-2-naphthacenecarboxamide; tetracycline (Compound L) ##STR16## (12) N-methyl-N-(3-phenyl-2-propenyl)-1-naphthalenemethanamine; naftifine (Compound M) ##STR17## (13) 2-(p-methoxybenzyl)-3,4-pyrrolidinediol-3-acetate; anisomycin (Compound N) ##STR18## The foregoin compounds hereinafter may be identified by the generic or USAN name or by "Compound" followed by the letter designations. As previously indicated, the combinations which are noteworthy are those combinations of 25-azasterol compounds with inhibitors of enzymes in the biochemical pathway in lanosterol synthesis. These combinations show fungicidal properties as well as synergistic antifungal properties. Especially noteworthy are combinations of 25-azasterol compounds with inhibitors of mevalonic acid synthesis, i.e., a 25-azasterol compound with a HMG-CoA synthase inhibitor compound; a 25-azasterol compound with a .beta.-ketothiolase inhibitor compound and a 25-azasterol compound with a HMG-CoA reductase inhibitor compound. Particularly useful specific combinations are 25-azacholesterol as the 25-azasterol compound with 11-(3-(hydroxymethyl)-4-oxo-2-oxetanyl)-7-methyl-2,4-undecadienoic acid as (Compound A) the HMG-CoA synthase inhibitor compound, with 5-(1-hydroxy-2,4,6-heptatriynyl)-2-oxo-b 1,3-dioxalane-4-heptanoic acid (Compound B) as the .beta.-ketothiolase inhibitor compound, and with 1,2,3,7,8,8a-hexahydro-3,7-dimethyl 8-[2-(tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1-naphthalenyl-2-but anoate (Compound E; lovastatin) as the HMG-CoA reductase inhibitor compound. Also noteworthy are combinations of 25-azasterol compounds with squalene epoxidase inhibitor compounds such as (E)-N-(6,6-dimethyl-2-hepten-4-ynyl)-N-methyl-1-naphthalene-methylamine (Compound D; terbinafine) and N-methyl-N-(3 phenyl-2-propenyl)-1-naphthalenemethanamine hydrochloride (Compound M; naftifine). Especially useful of the latter is the combination of 25-azacholesterol and terbinafine. The synergistic antifungal and fungicidal combinations of the present invention are effective in the treatment of mycotic infections caused by such fungal organisms as those of the Candida species, for example, C. albicans, C. parapsilosis, C. tropicalis, C. pseudotropicalis, C. krusei, C. rugosa, C. quilliermondii, C. stellatoidea; those of the Aspergillus species such as A. fumigatus; and other disease-causing fungi such as Cryptococcus neoformans; Torulopsis glabrata; Rhizopus rhizopodiformis; Coccidioides immitis; Sporothrix schenkii; Histoplasma capsulatum; and Blastomyces dermatitidis. The method for potentiating the antifungal effectiveness of an antifungal compound comprising employing a subfungistatic amount of an antifungal compound together with a 25-azasterol compound which if it has some fungistatic property is employed at a concentration not inhibitory to the growth of fungi. The potentiated combination may then be employed to control fungal growth by administering to or directing to a site where control of fungi is desired, an antifungally effective amount of a composition comprising a 25-azasterol compound in an amount non-inhibitory to fungal growth and an antifungal compound. Alternatively, each compound may be administered sequentially. The method is particularly directed to treating subjects with mycotic infections to control fungal growth and the disease caused by fungi by comprising administering to said subjects an antifungally effective amount of a composition comprising a 25-azasterol compound together with an antifungal agent. The application may be made at a site remote from that of the infection such as would be the case with oral or parenteral administration, or directly at the site infected with fungi. The agents may be administered with or without, preferably with, a pharmaceutically acceptable carrier in the amounts hereinafter set forth. By the administration of the amounts of the agents as hereinafter set forth, a potentiated or synergistic fungistatic interaction of the drugs is achieved which is wholly unexpected. With some combinations, namely, azasterols with lanosterol synthesis inhibitors, a fungicidal effect also may be achieved, usually at higher concentrations. With combinations which have a fungicidal effect, the invention is directed to killing fungi by administering to the site infected with fungi, the fungicidal combination. It is further directed to treating subjects with mycotic infection to eradicate, the disease-causing fungi by administering a fungicidal composition comprising an azasterol compound and a HMG-CoA synthase inhibitor, an azasterol compound and a .beta.-ketothiolase inhibitor an azasterol compound and a HMG-CoA reductase inhibitor or an azasterol compound and a squalene epoxidase inhibitor. As with fungistatic compositions, the application may be made at a site remote from the infection as in oral or parentaral administration, or may be made directly at the site of infection. The administration may be made with or without a carrier and in amounts as hereinafter detailed. By these operations, a wholly unexpected eradication of fungi has been achieved. The effectiveness of the combination generally depends on the original susceptibility of the particular organism or strain of organism to the fungistat to be employed in combination with the azasterol compound. Thus, although the azasterol compound has a potentiating effect on fungistats generally, the apparent greater effect of particular combinations may depend on the original susceptibility of the organism to the unmodified fungistat. In the case of fungistats which are known to be lanosterol synthesis inhibitors, it has been found with these fungistats that not only a synergistic fungistatic combination is obtained but also a fungicidal combination is obtained. The potentiation of the antifungal properties of an antifungal agent by coadministering a 25-azasterol compound and the production of a synergistic antifungal as well as in many instances of a fungicidal effect may be illustrated by the in vitro interaction studies for the determination of activity. In these tests against representative fungal organisms known to be the causative agent of mycotic infections, including Candida albicans, other Candida species, and a number of other fungi, synergistic antifungal properties have been demonstrated with a 25-azasterol compound together with various fungistatic lanosterol synthesis inhibitor compounds. |
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