Real-Time Imaging of the Azole Class of Antifungal Drugs in Live Candida Cells

• DOI: 10.1021/acschembio.7b00339
• Publication Date: 2017-05-04T21:13:24Z
• Authors: Raphael I. Benhamou; Maayan Bibi; Kfir B. Steinbuch; Hamutal Engel; Maayan Levin; Yael Roichman; Judith Berman; Micha Fridman
• Keywords: Azole; Lanosterol; Triazole; Antifungal drugs

Azoles are the most commonly used class of antifungal drugs, yet where they localize within fungal cells and how imported remain poorly understood. Azole antifungals target lanosterol 14α-demethylase, a cytochrome P450, encoded by ERG11 in Candida albicans, prevalent pathogen. We report synthesis fluorescent probes that permit visualization azoles live cells. Probe 1 is dansyl dye-conjugated azole, probe 2 Cy5-conjugated azole. Docking computations indicated each can occupy active site P450. Like azole drug fluconazole, not effective against mutant lacks In contrast, ketoconazole retained some activity mutants lacking implying both act more than one target. Both colocalized with mitochondria, as determined fluorescence microscopy MitoTracker dye. Thus, these useful molecular tools lead to detailed information about localization important drugs.