Interaction of amphotericin B and its low toxic derivative, N-methyl-N-D-fructosyl amphotericin B methyl ester, with fungal, mammalian and bacterial cells measured by the energy transfer method.
Amphotericin B (AMB) derivative, N-methyl-N-D-fructosyl amphotericin B methyl ester (MFAME) retains the broad antifungal spectrum and potency of the parent antibiotic, whereas its toxicity towards mammalian cells is reduced by about two orders of magnitude. The purpose of this work was to find out whether the differences observed in the toxicity of MFAME and native AMB are due to the differential drugs affinity to fungal and mammalian cell membranes. Comparative studies on AMB and MFAME biological activity and their affinity to fungal, mammalian and bacterial cells were performed. The interaction of AMB and MFAME with cells have been studied by fluorescence method based on the energy transfer between membrane fluorescent probe (donor) and the polyenic chromophore of the antibiotic (acceptor) simultaneously present in the cell membrane. The amount of the antibiotic bound to cells was indicated by the extent of fluorescence quenching of 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) or 1,6-diphenyl-1,3,5-hexatriene (DPH) by polyenic chromophore of the antibiotic. The results obtained indicate that binding extent and characteristics for both antibiotics are comparable in the three types of cells studied. Dramatically lower toxicity of MFAME as compared to AMB towards mammalian cells is not related to the antibiotic-cell affinity, but rather to different consequences of these interactions for cells, reflected in membrane permeabilization. MFAME is definitely less effective than parent AMB in the permeabilizing species formation in mammalian cell membrane.
Szlinder-Richert
Joanna
J
Department of Pharmaceutical Technology and Biochemistry, Gdansk University of Technology, 11/12, Narutowicza St, 80-952 Gdansk, Poland.
Cybulska
Barbara
B
Grzybowska
Jolanta
J
Bolard
Jacques
J
Borowski
Edward
E
D000666:Amphotericin B / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology; D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology; D002176:Candida albicans / Q000187:drug effects / Q000378:metabolism*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions / Q000502:physiology; D004735:Energy Transfer / Q000187:drug effects / Q000502:physiology*; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000706:statistics & numerical data
D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; C108557:N-methyl-N-fructopyranosylamphotericin B methyl ester; D000666:Amphotericin B