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WO1997048278A1 - Activite antifongique des spongistatines - Google Patents

Activite antifongique des spongistatines Download PDF

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Publication number
WO1997048278A1
WO1997048278A1 PCT/US1997/010200 US9710200W WO9748278A1 WO 1997048278 A1 WO1997048278 A1 WO 1997048278A1 US 9710200 W US9710200 W US 9710200W WO 9748278 A1 WO9748278 A1 WO 9748278A1
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WO
WIPO (PCT)
Prior art keywords
spongistatin
active ingredient
candida
spongistatins
yeast
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Ceased
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PCT/US1997/010200
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English (en)
Inventor
George R. Pettit
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University of Arizona
Arizona's Public Universities
Arizona State University ASU
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University of Arizona
Arizona's Public Universities
Arizona State University ASU
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Filing date
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Priority claimed from US08/876,407 external-priority patent/US5883120A/en
Application filed by University of Arizona, Arizona's Public Universities, Arizona State University ASU filed Critical University of Arizona
Publication of WO1997048278A1 publication Critical patent/WO1997048278A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom

Definitions

  • This invention relates generally to the discovery of very potent antifungal activity for spongistatin 1 and the related spongistatins (2 through 7, inclusive) which are hereinafter described in detail. More particularly, this invention relates to the discovery of new and unexpected antifungal properties for a series of macrocyclic lactone polyethers (spongistatins) which have heretofore been isolated from the marine sponges Spongia sp. and Spirastrella spini spirulifera . These unexpected properties include inhibition of the growth of the opportunistic fungi Candida albi cans and Cryptococc ⁇ s neoformans in disk diffusion and broth macrodilution assays.
  • spongistatins macrocyclic lactone polyethers
  • spongistatin 1 was fungicidal for C. albicans and C. neoformans . At 35°C, spongistatin 1 appeared to induce an altered germ tube morphology in C. albicans . This research was funded in part by
  • Marine organisms are proving to be very productive sources of new therapeutic agents. Indeed, some of today's most promising anticancer drugs in clinical or preclinical trials have been isolated from marine invertebrates or their associated microbes. (Flam, F. 1994. Chemical prospectors scour seas for promising drugs. Science 266:1324-1325; Pettit, G.R. 1994. Marine animal and terrestrial plant anticancer constituents. Pure & Appl. Chem. 66:2271-2281).
  • the spongistatins are a family of macrocyclic lactone polyethers recently isolated from marine Porifera . Spongistatins 1-3 were discovered in Eastern Indian Ocean Spongia sp. (Pettit, G.R., Z.A.
  • FIG. 1 is the killing kinetics of spongistatin 1 with no drug (squares) and at IX (circles) and 2X (triangles) the MIC for C. albicans.
  • FIG. 2 is the killing kinetics of spongistatin 1 with no drug (squares) and at IX (circles) and 2X (triangles) the broth macrodilution MIC for C. neojforiTians.
  • a source of spongistatins 1-7 was located at the Arizona State University Cancer Research Institute, Tempe, Arizona.
  • Antimicrobial activity was assayed by both broth macrodilution and disk susceptibility tests according to methods established by the National Committee for Clinical Laboratory Standards. (National Committee for Clinical Laboratory Standards. 1990. Approved standard M2-A4. Performance standards for antimicrobic disk susceptibility tests. National Committee for Clinical Laboratory Standards, Villanova, PA; National Committee for Clinical Laboratory Standards. 1994. Reference method for broth dilution antifungal susceptibility testing of yeasts; Tentative standard. NCCLS document M27-T. National Committee for Clinical Laboratory Standards, Villanova, PA) .
  • gonorrhoeae or 50 ⁇ l ⁇ E. coli , C. albicans, C. neoformans spread on the appropriate plates. Excess moisture was allowed to absorb for 10 minutes before applying disks.
  • spongi ⁇ statins 1-7 were reconstituted in sterile dimethyl- sulfoxide, and two-fold dilutions applied to sterile 6 mm disks. Disks were dried at 37*C, and applied to inoculated plates. Test plates of E. coli , S. aureus and E. faecalis were incubated at 37°C, N.
  • gonorrhoeae at 37°C with 5% C0 2
  • C. albi cans and C. neoformans at 25"C Zones of inhibition were recorded after 16 hours for bacterial cultures, and 42 hours for fungal cultures.
  • the MIC was defined as the lowest concentration of drug resulting in a clear zone of growth inhibition.
  • spongistatin 1 was the most potent compound, and this is consistent with the relative cytotoxicities of the spongistatins against human cancer cell lines, (Pettit, G.R., Z.A. Cichacz, F. Gao, CL. Herald, and M.R. Boyd. 1993.
  • Spongistatin 1 a highly cytotoxic, sponge-derived, marine natural product that inhibits mitosis, microtubule assembly, and the binding of vinblastine to tubulin. Molec. Pharmacol. 44:757- 766; Bai, R. , G.F. Taylor, Z.A. Cichacz, CL. Herald, J.A. Kepler, G.R. Pettit, and E. Hamel. 1995.
  • Spongistatin 1 is the most abundant [3.4 x 10 "7 % yields (Pettit, G.R., Z.A. Cichacz, CL. Herald, M.R. Boyd, J.M. Schmidt, and J.N.A. Hooper. 1993. Isolation and structure of spongistatin 1. J. Org. Chem. 58:1302-1304)] of the lactone polyethers isolated from Spongia sp. and Spirastrella spinispirulifera .
  • spongistatin 1 The antifungal activity of spongistatin 1 was also tested by the broth macrodilution assay (National Committee for Clinical Laboratory Standards. 1994. Reference method for broth dilution antifungal susceptibility testing of yeasts; Tentative standard. NCCLS document M27-T. National Committee for Clinical Laboratory Standards, Villanova, PA) . C. albi cans and C. neoformans were maintained at 35°C on YM agar, and inocula prepared as recommended in document M27-T.
  • Tests were performed in sterile 12x75 mm plastic tubes containing two-fold dilutions of spongistatin 1 (reconstituted in DMSO) in 0.165 M morpholine- propanesulfonic acid buffered RPMI 1640 medium (pH 7.0). Tubes were incubated without agitation at 35"C Minimum inhibitory concentrations (MICs) were determined after 48 hours for C. albicans and 72 hours for C. neoformans. The MIC was defined as the lowest concentration of spongistatin 1 that inhibited all visible growth of the test organism. Minimum fungicidal concentrations (MFCs) were determined by subculturing 0.1 ml from each tube with no visible growth in the MIC broth macrodilution series onto a drug-free YM plate.
  • MFCs Minimum fungicidal concentrations
  • the plates were incubated at 35°C for 24 hours for C. albicans, and 48 hours for C. neoformans .
  • the MFC was defined as the lowest concentration of 8278 PC17US97/10200
  • spongistatin 1 inhibited fungi in two radically different susceptibility assays, disk diffusion and broth macrodilution, may be predictive of in vivo effectiveness.
  • spongistatin 1 was fungicidal for C. albicans and C. neoformans at 2X MIC ( Figures 1 and 2) .
  • Spongistatin 1 at 2X MIC caused a 1 log 10 reduction in the cfu/ml of C. albi cans after 6 hours of incubation, and after 12 hours of incubation for C. neoformans.
  • After 24 hours at 2X MIC there was a 2 log 10 reduction in C. albicans cfu/ml, and no viable C. neoformans .
  • yeast cells were the predominant morphologic form in untreated C. albicans cultures. However, from 6 hours on elongated structures predominated in 2X MIC treated cultures. The most abundant elongated forms more closely resembled germ tubes than hyphae. No obvious morphological alterations in spongistatin 1 treated C. neoformans were observed.
  • Tubulin the major component of microtubules, is the target of many naturally occurring compounds that cause cells to arrest in mitosis.
  • Hamel E. 1990. Interactions of tubulin with small ligands, p. 89-191.
  • J. Avila ed.
  • Microtubule Proteins. CRC Press Boca Raton, FL
  • marine mollusks and sponges containing novel antimitotic agents have been described. (Bai, R. , S.J. Friedman, G.R. Pettit, and E. Hamel. 1992.
  • Dolastatin 15 a potent antimitotic depsipeptide derived from Dolabella auri cularia : interaction with tubulin and effects on cellular microtubules. Biochem. Pharmacol. 43:2637-2645; Bai, R. , K.D. Paull, CL. Herald, L. Malspeis, G.R. Pettit, and E. Hamel. 1991. Halichondrin B and homohalichondrin B, marine natural products binding in the Vinca domain of tubulin: discovery of tubulin-based mechanism of action by analysis of differential cytotoxicity data. J. Biol. Chem. 266:15882-15889). In mammalian cells, spongistatin 1 has also been shown to be antimitotic.
  • spongistatin 1 caused the accumulation of cells arrested in mitosis, and the disappearance of intracellular microtubules.
  • Spongistatin 1 a highly cytotoxic, sponge- derived, marine natural product that inhibits mitosis, microtubule assembly, and the binding of vinblastine to tubulin. Molec. Pharmacol. 44:757- 766) .
  • Spongistatin 1 also inhibited the glutamate- induced polymerization of purified bovine brain tubulin. (Bai, R.
  • spongistatins 1-7 The structures of spongistatins 1-7, the macrocyclic lactone polyethers isolated from Spongia sp. (spongistatins 1-3) and Spirastrella spinispirulifera (spongistatins 4-7) are shown below:
  • R2 H
  • Spongistatins 1-7 were reconstituted in sterile dimethylsulfoxide (DMSO) immediately prior to all assays. DMSO alone had no detectable inhibitory effect on any of the tested microbes.
  • DMSO sterile dimethylsulfoxide
  • Yeast strains (listed in Table IV) were maintained by single colony transfer on Yeast Morphology (YM) agar (Difco) at 35°C ATCC#32354 is flucytosine resistant, ATCC#64124 is keto- conazole resistant and ATCC#42720 is amphotericin B resistant. Issatchenkia ori entalis was formerly classified as Candida krusei , and Rhodotorula mucilaginosa as Rhodotorula rubra (ATCC product literature) . Filamentous fungi (Table IV) were maintained on Potato Dextrose Agar (PDA) slants at 35°C
  • PDA Potato Dextrose Agar
  • Spongistatin 1 was screened against yeasts by the broth macrodilution assay according to the NCCLS (1994) . Antibiotic resistant strains were subcultured only once after acquisition to ensure no loss of antibiotic resistance. Yeasts were suspended and diluted as recommended to yield final inocula ranging from 0.5-2.5 x 10 3 cfu/ml. Tests were performed in sterile 12x75 mm plastic tubes containing two-fold dilutions of spongistatin 1 in 0.165 M morpholinepropanesulfonic acid (MOPS)- buffered RPMI 1640 medium (pH 7.0). One tube was left drug-free for a turbidity control.
  • MOPS morpholinepropanesulfonic acid
  • Tubes were incubated without agitation at 35°C MICs were determined after 48 hours for all yeast strains except Cryptococcus , which was read after 72 hours.
  • the MIC was defined as the lowest concentration of spongistatin 1 that inhibited all visible growth of the test organism.
  • MFCs Minimum fungicidal concentration
  • fumigatus R. and oligosporus were grown on PDA slants at 35 ⁇ C for 6 days. Fungal slants were covered with 1 ml sterile 0.85% NaCl, and suspensions made by gently probing the colonies with the tip of a sterile pipette. The resulting mixture of hyphal fragments and conidia or sporangiospores was withdrawn, transferred to a sterile clear microfuge tube, and heavy particles allowed to settle for 10 minutes.
  • the upper homogeneous suspension was transferred to a sterile microfuge tube, vortexed 15 s, adjusted spectro- photometrically, and diluted in sterile 0.165M MOPS buffered RPMI 1640 medium, pH 7.0, to yield final inocula ranging from 0.5-2.5 x 10 3 cfu/ml.
  • Susceptibility to spongistatin 1 was then determined by broth macrodilution assays as described above for the yeast cultures. MICs for the filamentous fungi were read after 24 hours, aliquots were aseptically removed for dilution plating and microscopy, and MFCs read 24 hours later.
  • spongistatins 1-7 did not inhibit the tested bacterial strains. However, all of the spongi ⁇ statins inhibited growth of the opportunistic fungi C. albicans and C. neoformans in disk diffusion assays (Table III) .
  • Spongistatin 1 is the most abundant (typical yield 5 mg/400 kg of wet sponge) of the lactone polyethers isolated from Hyrtios sp. and S. spinispirulifera . In broth macrodilution tests, spongistatin 1 was fungicidal for all yeast and filamentous fungi examined, including flucytosine-resistant C. albicans , ketoconazole- resistant C. albi cans and amphotericin B-resistant C. l usi taniae (Table IV).
  • yeast cells were the predominant morphologic form in untreated C. albicans cultures. However, from 6 hours on elongated structures predominated in 2X MIC treated cultures. The most abundant elongated forms more closely resembled germ tubes than hyphae. No obvious morphological alterations in spongistatin 1-treated C. neoformans were observed.
  • Spongistatin 1 6.25-12 .5 6.25-12.5 Spongistatin 2 25-50 50-100 Spongistatin 3 25-50 50-100 Spongistatin 4 12. 5-25 6.25-12.5 Spongistatin 5 25-50 6.25-12.5 Spongistatin 6 25-50 12.5-25 Spongistatin 7 6. 25-12 . 5 12.5-25
  • Table IV Antifungal activity of spongistatin 1 in the broth macrodilution assay.
  • compositions are believed useful in the treatment of one or more fungal infections, such as Aspergill osi s , Candidiasis or thrush, internal infections such as cryptococcosis , epidermal infections, infections caused by antibiotic resistant fungi and the like. Similar fungal infections are enumerated in the AMA Home Medical Encyclopedia published by Random House, Inc. 1989.
  • the dosage administered will be dependent upon the identity of the fungus; the location of the fungal infection; the type of host involved; the nature of concurrent treatment, if any; and the frequency of treatment specified.
  • dosage levels of the administered active ingredients are: intravenous, 0.1 to about 200 mg/kg; orally, 5 to about 1000 mg/kg of host body weight.
  • an active ingredient can be present in the compositions of the present invention for localized use about the cutis, intranasally, pharyngolaryngeally, bronchially, intravaginally, or ocularly in a concentration of from about 0.01 to about 50% w/w of the composition; preferably about 1 to about 20% w/w of the composition; and for parenteral use in a concentration of from about 0.05 to about 50% w/v of the composition and preferably from about 5 to about 20% w/v.
  • compositions of the present invention are preferably presented for administration to humans and animals in salves and ointments for topical application although unit dosage forms, such as tablets, capsules, pills, powders, suppositories, sterile parenteral solutions or suspensions, sterile non-parenteral solutions or suspensions, lozenges and the like, containing suitable quantities of an active ingredient.
  • powders are prepared quite simply by comminuting the active ingredient to a suitably fine size and mixing with a similarly comminuted diluent.
  • the diluent can be an edible carbohydrate material such as lactose or starch.
  • Advanta ⁇ geously, a sweetening agent or sugar is present as well as a flavoring oil.
  • Capsules are produced by preparing a powder mixture as hereinbefore described and filling into formed gelatin sheaths.
  • a lubricant such as talc, magnesium stearate, calcium stearate and the like is added to the powder mixture before the filling operation.
  • Soft gelatin capsules are prepared by machine encapsulation of a slurry of active ingredients with an acceptable vegetable oil, light liquid petrolatum or other inert oil or triglyceride. Tablets are made by preparing a powder mixture, granulating or slugging, adding a lubricant and pressing into tablets.
  • the powder mixture is prepared by mixing an active ingredient, suitably comminuted, with a diluent or base such as starch, lactose, kaolin, dicalcium phosphate and the like.
  • the powder mixture can be granulated by wetting with a binder such as corn syrup, gelatin solution, methylcellulose solution or acacia mucilage and forcing through a screen.
  • the powder mixture can be slugged, i.e., run through the tablet machine and the resulting imperfectly formed tablets broken into pieces (slugs) .
  • the slugs can be lubricated to prevent sticking to the tablet-forming dies by means of the addition of stearic acid, a stearic salt, talc or mineral oil.
  • the lubricated mixture is then compressed into tablets.
  • the tablet can be provided with a protective coating consisting of a sealing coat or enteric coat of shellac, a coating of sugar and methylcellulose and polish coating of carnauba wax.
  • Fluid unit dosage forms for oral adminis ⁇ tration such as in syrups, elixirs and suspensions can be prepared wherein each teaspoonful of composition contains a predetermined amount of an active ingredient for administration.
  • the water- soluble forms can be dissolved in an aqueous vehicle together with sugar, flavoring agents and preservatives to form a syrup.
  • An elixir is prepared by using a hydroalcoholic vehicle with suitable sweeteners together with a flavoring agent.
  • Suspensions can be prepared of the insoluble forms with a suitable vehicle with the aid of a suspending agent such as acacia, tragacanth, methylcellulose and the like.
  • fluid unit dosage forms are prepared utilizing an active ingredient and a sterile vehicle, water being preferred.
  • the active ingredient can be either suspended or dissolved in the vehicle.
  • the active ingredient can be dissolved in a suitable vehicle for injection and filter sterilized before filling into a suitable vial or ampule and sealing.
  • adjuvants such as a local anesthetic, preservative and buffering agents can be dissolved in the vehicle.
  • Parenteral suspensions are prepared in substantially the same manner except that an active ingredient is suspended in the vehicle instead of being dissolved and sterilization cannot be accomplished by filtration.
  • the active ingredient can be sterilized by exposure to ethylene oxide before suspending in the sterile vehicle.
  • a surfactant or wetting agent is included in the composition to facilitate uniform distribution of the active ingredient.
  • vaginal routes can be utilized particularly in the treatment of candidiasis (thrush) .
  • An active ingredient can be administered by means of a suppository.
  • a vehicle which has a melting point at about body temperature or one that is readily soluble can be utilized.
  • cocoa butter and various polyethylene glycols (Carbowaxes) can serve as the vehicle.
  • the active ingredients can be packaged in a pressurized aerosol container together with a gaseous or liquified propellant, for example, dichlorodifluoromethane, carbon dioxide, nitrogen, propane, and the like, with the usual adjuvants such as cosolvents and wetting agents, as may be necessary or desirable.
  • a gaseous or liquified propellant for example, dichlorodifluoromethane, carbon dioxide, nitrogen, propane, and the like
  • the active ingredient will be delivered to the site as an ointment or salve which will comprise water and oil emulsion as the principal carrier.
  • Other conventional ingredients when conditions and aesthetics dictate, include petrolatum and mineral oil, lipophilic solubilizers such as polyethylene glycol, carbowax, moisturizers such as lanolin and fragrance.
  • unit dosage form refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical diluent, carrier or vehicle.
  • the specifications for the novel unit dosage forms of this invention are dictated by and are directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitation inherent in the art of compounding such an active material for therapeutic use in humans, as disclosed in this specification, these being features of the present invention.
  • suitable unit dosage forms in accord with this invention are tablets, capsules, troches, suppositories, powder packets, wafers, cachets, teaspoonfuls, tablespoonfuls, dropperfuls, ampules, vials, segregated multiples of any of the foregoing, and other forms as herein described.
  • the active ingredients to be employed as antifungal agents can be easily prepared in such unit dosage form with the employment of pharmaceutical materials which themselves are available in the art and can be prepared by established procedures.
  • the following preparations are illustrative of the preparation of the unit dosage forms of the present invention, and not as a limitation thereof.
  • Several dosage forms were prepared embodying the present invention. They are shown in the following examples in which the notation "active ingredient” signifies one of the compound designated as "spongistatin”; specifically, spongistatin 1 through spongistatin 7, inclusive.
  • the active ingredient finely divided by means of an air micronizer, is added to the other finely powdered ingredients, mixed thoroughly and then encapsulated in the usual manner.
  • the foregoing capsules are useful for treating a fungal disease by the oral administration of one or two capsules one to four times a day.
  • capsules are similarly prepared containing an active ingredient in 50, 250 and 500 mg amounts by substituting 50 g, 250 g and 500 g of an active ingredient for the 200 g used above.
  • COMPOSITION "B” Soft Gelatin Capsules One-piece soft gelatin capsules for oral use, each containing 200 mg of an active ingredient, finely divided by means of an air micronizer, are prepared by first suspending the compound in 0.5 ml of corn oil to render the material capsulatable and then encapsulating in the above manner.
  • the foregoing capsules are useful for treating a fungal disease by the oral administration of one or two capsules one to four times a day.
  • COMPOSITION "C” Tablets One thousand tablets, each containing 200 mg of an active ingredient, are prepared from the following types and amounts of ingredients:
  • the active ingredient finely divided by means of an air micronizer, is added to the other ingredients and then thoroughly mixed and slugged.
  • the slugs are broken down by forcing them through a Number Sixteen screen.
  • the resulting granules are then compressed into tablets, each tablet containing 200 mg of the active ingredient.
  • the foregoing tablets are useful for treating a fungal disease by the oral administration of one or two tablets one to four times a day.
  • tablets are similarly prepared containing an active ingredient in 250 mg and 100 mg amounts by substituting 250 g and 100 g of an active ingredient for the 200 g used above.
  • COMPOSITION "D" Oral Suspension One liter of an aqueous suspension for oral use, containing in each teaspoonful (5 ml) dose, 50 mg of an active ingredient, is prepared from the following types and amounts of ingredients: Active ingredient, micronized 10 g
  • Deionized water q.s. 1000 ml
  • the citric acid, benzoic acid, sucrose, tragacanth and lemon oil are dispersed in sufficient water to make 850 ml of suspension.
  • the active ingredient finely divided by means of an air micronizer, is stirred into the syrup unit uniformly distributed. Sufficient water is added to make 1000 ml.
  • composition so prepared is useful for treating a fungal disease at a dose of 1 teaspoonful (15 ml) three times a day.
  • COMPOSITION "E" Parenteral Product One liter of a sterile aqueous suspension for parenteral injection, containing 30 mg of an active ingredient in each milliliter for treating a fungal disease, is prepared from the following types and amounts of ingredients:
  • composition so prepared is useful for treating a fungal disease at a dose of 1 milliliter (1ml) three times a day.
  • COMPOSITION "F" Vaginal Suppository One thousand suppositories, each weighing 2.5 g and containing 200 mg of an active ingredient are prepared from the following types and amounts of ingredients:
  • Polyethylene glycol #4000, q.s. 2,500 g The active ingredient is finely divided by means of an air micronizer and added to the propylene glycol and the mixture passed through a colloid mill until uniformly dispersed.
  • the polyethylene glycol is melted and the propylene glycol dispersion is added slowly with stirring.
  • the suspension is poured into unchilled molds at 40°C
  • the composition is allowed to cool and solidify and then removed from the mold and each suppository foil wrapped.
  • the foregoing suppositories are inserted vaginally for treating candidiasi s (thrush) .
  • COMPOSITION "G" Intranasal Suspension One liter of a sterile aqueous suspension for intranasal instillation, containing 20 mg of an active ingredient in each milliliter, is prepared from the following types and amounts of ingredients:
  • composition so prepared is useful for treating a fungal disease, by intranasal instillation of 0.2 to 0.5 ml given one to four times per day.
  • An active ingredient can also be present in the undiluted pure form for use locally about the cutis, intranasally, pharyngolaryngeally, bronchially, or orally.
  • COMPOSITION "H" Powder Five grams of an active ingredient in bulk form is finely divided by means of an air micronizer. The micronized powder is placed in a shaker-type container. The foregoing composition is useful for treating a fungal disease, at localized sites by applying a powder one to four times per day.
  • micronized powder is divided into individual doses of 200 mg and packaged.
  • the foregoing powders are useful for treating a fungal disease, by the oral administration of one or two powders suspended in a glass of water, one to four times per day.
  • One hundred grams of an active ingredient in bulk form is finely divided by means of an air micronizer.
  • composition is useful for treating a fungal disease, by the inhalation of 300 mg one to four times a day.
  • COMPOSITION "K” Ointment One hundred grams of an active ingredient in bulk form is finely divided by means of an air micronizer. The micronized powder is them admixed into a water and oil emulsion with the addition of suitable moisturizers and fragrances as desired.
  • ointment is useful for treating a fungal disease by one topical application of the ointment on the affected area as needed, preferably at least twice a day. From the foregoing, it becomes readily apparent that a new and useful antifungal agent and new and useful antifungal preparations have been herein described and illustrated which fulfill the aforestated object in a remarkably unexpected fashion. It is further believed that the spongistatin can be utilized as probes to examine fungal cell biochemistry including morphogenesis and cell division. It is, of course, understood that such modifications, alterations and adaptions as will readily occur to the artisan confronted with this disclosure are intended within the spirit of the present invention which is limited only by the scope of the claims appended hereto.

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Abstract

La présente invention concerne généralement le domaine des agents anti-fongiques et une préparation pharmaceutique contenant ces agents qui semblent être potentiellement utiles dans le domaine de la thérapie antifongique. Plus particulièrement, l'invention concerne la découverte de propriétés remarquablement inespérées pour une série de polyéthers de lactone macrocycliques (spongistatines) qui sont dérivés des éponges marines Spongia sp. et Spirastrella spinispirulifera et qui ont inhibé la croissance de levures et de champignons filamenteux lors d'analyses de diffusion sur disque et de macrodilution en bouillon de culture (pour la spongistatine 1). Des formes de posologie thérapeutique sont également décrites pour le traitement d'infections provoquées par Candida albicans, Cryptococcus neoformans, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Rhodotorula mucilaginosa, Aspergillus fumigatus, Rhizopus ologosporus, et d'autres levures et champignons filamenteux renfermant des souches résistant à l'amphotéricine B, au kétoconazole, et à la flucytosine. L'activité fongicide de la spongistatine 1 a été confirmée lors d'études cinétiques de mise à mort, où la mise à mort de Candida albicans est intervenue en moins de 6 heures. Les spongistatines sont prometteuses comme agents antifongiques potentiels et comme sondes pour l'étude de la morphogénèse fongique et de la division nucléaire.
PCT/US1997/010200 1996-06-18 1997-06-17 Activite antifongique des spongistatines Ceased WO1997048278A1 (fr)

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US08/876,407 US5883120A (en) 1997-06-16 1997-06-16 Antifungal activity of the spongistatins

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7473433B2 (en) 2000-12-21 2009-01-06 Nektar Therapeutics Pulmonary delivery of polyene antifungal agents
US11174291B2 (en) 2015-02-13 2021-11-16 Arizona Board Of Regents On Behalf Of Arizona State University Silstatin compounds

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US5328929A (en) * 1993-07-02 1994-07-12 Arizona Board Of Regents Isolation and structure of spongistatin 2, spongistatin 3, spongistatin 4 and spongistatin 6
US5393897A (en) * 1993-07-02 1995-02-28 Arizona Board Of Regents Acting On Behalf Of Arizona State University Isolation and structure of spongistatins 5,7,8 and 9
US5436400A (en) * 1993-01-19 1995-07-25 Arizona Board Of Regents Isolation and structure of spongistatin 1

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
US5436400A (en) * 1993-01-19 1995-07-25 Arizona Board Of Regents Isolation and structure of spongistatin 1
US5328929A (en) * 1993-07-02 1994-07-12 Arizona Board Of Regents Isolation and structure of spongistatin 2, spongistatin 3, spongistatin 4 and spongistatin 6
US5393897A (en) * 1993-07-02 1995-02-28 Arizona Board Of Regents Acting On Behalf Of Arizona State University Isolation and structure of spongistatins 5,7,8 and 9

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7473433B2 (en) 2000-12-21 2009-01-06 Nektar Therapeutics Pulmonary delivery of polyene antifungal agents
US11174291B2 (en) 2015-02-13 2021-11-16 Arizona Board Of Regents On Behalf Of Arizona State University Silstatin compounds

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