title: Antifungal activity of eupolauridine and its action on DNA topoisomerases.
abstract: The azafluoranthene alkaloid eupolauridine has previously been shown to have in vitro antifungal activity and selective inhibition of fungal topoisomerase I. The present study was undertaken to examine further its selectivity and mode of action. Eupolauridine completely inhibits the DNA relaxation activity of purified fungal topoisomerase I at 50 microg/ml, but it does not stabilize the cleavage complex of either human or fungal topoisomerase I. Cleavage complex stabilization is the mode of action of topoisomerase I targeting drugs of the camptothecin family. Also, unlike camptothecin, eupolauridine does not cause significant cytotoxicity in mammalian cells. To determine if the inhibition of topoisomerase I is the principal mode of antifungal action of eupolauridine, Saccharomyces cerevisiae strains with alterations in topoisomerase genes were used in clonogenic assays. The antifungal activity of eupolauridine was not diminished in the absence of topoisomerase I; rather, the cells lacking the enzyme were more sensitive to the drug. Cell-killing activity of eupolauridine was also more pronounced in cells that overexpressed topoisomerase II. In vitro assays with the purified yeast enzyme confirmed that eupolauridine stabilized topoisomerase II covalent complexes. These results indicate that a major target for fungal cell killing by eupolauridine is DNA topoisomerase II rather than topoisomerase I, but does not exclude the possibility that the drug also acts against other targets.
authors.affiliation: National Center for Natural Products Research, Department of Pharmacognosy, The University of Mississippi, University, Mississippi 38677, USA.
authors.forename: Shabana I
authors.initials: SI
authors.lastname: Khan
authors.forename: Alison C
authors.initials: AC
authors.lastname: Nimrod
authors.forename: Mohammed
authors.initials: M
authors.lastname: Mehrpooya
authors.forename: John L
authors.initials: JL
authors.lastname: Nitiss
authors.forename: Larry A
authors.initials: LA
authors.lastname: Walker
authors.forename: Alice M
authors.initials: AM
authors.lastname: Clark
mesh_terms: D000470:Alkaloids / Q000494:pharmacology*; D000935:Antifungal Agents / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects / Q000235:genetics; D002384:Catalysis; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media; D004271:DNA, Fungal / Q000737:chemistry / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005449:Fluorenes; D006801:Humans; D007192:Indenes; D008826:Microbial Sensitivity Tests; D009287:Naphthyridines; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D014407:Tumor Cells, Cultured
chemical_list: D000470:Alkaloids; D000935:Antifungal Agents; D003470:Culture Media; D004271:DNA, Fungal; D004791:Enzyme Inhibitors; D005449:Fluorenes; D007192:Indenes; D009287:Naphthyridines; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; C054957:eupolauridine