Patent GB-1349633-A (GB)
Title: Antifungal agents
IPC: C12P1/04
Filed: 1973-01-18
Abstract: 1349633 Antifungal antibiotics WARNERLAMBERT CO 18 Jan 1973 [15 Feb 1972] 2591/73 Heading C2A The novel antifungal antibiotics Acid F and Acid S, having the following characteristics in common: Empirical formula: C 28 H 42 O 6 ; mol. weight 474; and infra-red spectrum having absorption peaks at 870, 965, 1063, 1255, 1388, 1453, 1663, 1710, 2950 and 3400 cm.<SP>-1</SP>; and being distinguished from each other by their specific rotations and Rf values, viz. Acid F: [α]<SP>#25</SP> D =+74 (C=0À8, CHCl 3 ); Rf=0À68 (on silica gel developing with EtAc: isopropanol: H 2 O, 85 : 10 : 5); Acid S: [α]25 D = +36 (C=0À7, CHCl 3 ); Rf=0À56 (on silica gel developing with EtAc: isopropanol: H 2 O, 85 : 10 : 5) are produced by aerobically cultivating Polyangium cellulosum var. fulvum (ATCC. 25532) in an aqueous nutrient medium containing assimilable sources of carbon and nitrogen, filtering the culture broth, extracting the filtrate with a water-immiscible non-polar solvent, concentrating the extract and then filtering the concentrate, adsorbing the filtrate on a column of activated magnesium silicate, washing the column with a non-polar solvent and then eluting the mixed acids from the column with a polar solvent. The eluate may be evaporated to dryness to give a crude product containing 20- 30% of the mixed acids; or else concentrated, acidified, extracted with chloroform and the latter extract concentrated and purified by treatment with silica gel; the purified concentrate is then evaporated to dryness to give a pure mixture of the acids. The polar solvent is preferably methanol, while typical non-polar solvents are ethyl acetate and chloroform. Acids F and S are separated from each other by column chromatography on silica gel with ethyl acetate : isopropanol : water (85 : 10: 5). The aforesaid crude product from the evaporation of the polar solvent eluate is used for this separation step, the eluates from the column being monitored by thin layer silica gel chromatography with the aforesaid developing solvent. The fractions containing the individual antibiotics are then evaporated to dryness to give the separate Acid F and Acid S. The antibiotics Acid F and Acid S may be converted to their novel methyl esters, Ester F and Ester S, which are also antifungal antibiotics having in common the empirical formula: C 29 H 44 O 6 , and molecular weight 488. Their infra-red spectra are respectively: 860, 965, 1060, 1165, 1255, 1370, 1430, 1515, 1660, 1730, 2900 and 3350 cm.<SP>-1</SP>; and 860, 965, 1060, 1200, 1250, 1370, 1440, 1518, 1658, 1725, 2900 and 3400 cm.<SP>-1</SP>. Their respective Rf values on silica gel developing with EtAc : cyclohexane (4 : 1) are 0À78 and 0À56, which property provides a means for their separation from each other. They may be prepared by treating the Acids F and S, either before or after their separation from each other, with ethanolic-ethereal diazomethane for 15 mins. at ambient temperature. The resulting Esters F and S may be purified and/or separated from each other by chromatography on silica gel with ethyl acetate : cyclohexane (4 : 1), the individual elates then being evaporated to dryness. Reflux in IN NaOH hydrolyses the esters to the original acids. Acid F, Acid S and their derivatives Ester F and Ester S may be mixed with liquid or solid pharmaceutical carriers or diluents to provide antifungally active pharmaceutical compositions.