Fungal infections present a growing global public health
concern,
necessitating the development of novel antifungal drugs. However,
many potential antifungals, particularly the expelled potential active
agents (EPAAs), are often underestimated owing to their limitations
in cellular entry or expulsion by efflux pumps. Herein, we identified
68 EPAAs out of 2322 candidates with activity against a Candida albicans efflux pump-deficient strain and
no inhibitory activity against the wild-type strain. Using a novel
conjugation strategy involving benzamidine (BM) as a mitochondrion-targeting
warhead, we successfully converted EPAAs into potent antifungals against
various urgent-threat azole-resistantCandida strains. Among the obtained EPAA-BM conjugates, IS-2-BM (11) exhibited excellent antifungal activities and induced negligible
drug resistance. Furthermore, IS-2-BM prevented biofilm
formation, eradicated mature biofilms, and exhibited excellent therapeutic
effects in a murine model of systemic candidiasis. These findings
provide a promising strategy for increasing the possibilities of discovering
more antifungals.