Antifungal resistance due to upregulation of efflux pumps is prevalent in clinical Candida isolates. Potential efflux pump substrates (PEPSs), which are active against strains deficient in efflux pumps but inactive against wild-type strains, are usually missed in routine antifungal screening. Here we present a method for identification of PEPSs, and show that conjugation with mitochondria-targeting triphenylphosphonium cation (TPP+) can enhance or restore the compounds’ antifungal activity. The screening method involves co-culturing a wild-type C. albicans strain and a Cdr efflux pump-deficient strain, labelled with different fluorescent proteins. We identify several PEPSs from a library of natural terpenes, and restore their antifungal activity against wild-type and azole-resistant C. albicans by conjugation with TPP+. The most active conjugate (IS-2-Pi-TPP) kills C. albicans cells, prevents biofilm formation and eliminates preformed biofilms, without inducing significant resistance. The antifungal activity is accompanied by mitochondrial dysfunction and increased levels of intracellular reactive oxygen species. In addition, IS-2-Pi-TPP is effective against C. albicans in a mouse model of skin infection.

No abstract

One of the main azole-resistance mechanisms in Candida pathogens is the upregulation of drug efflux pumps, which compromises the efficacy of azoles and results in treatment failure. The combination of azole-antifungal agents with efflux pump inhibitors represents a promising strategy to combat fungal infection. High-throughput screening of 150 extracts obtained from endolichenic fungal cultures led to the discovery that the extract of Phialocephala fortinii exhibits potent activity for the reversal of azole resistance. From P. fortinii cultures, a total of 15 quinone derivatives, comprising 11 new derivatives and 4 known compounds, were obtained. Among these compounds, palmarumycin P3 (3) and phialocephalarin B (8) specifically modulate the expression of MDR1 to inhibit the activity of drug efflux pumps and therefore reverse azole resistance. The present study revealed Mdr1 targeting as an alternative mechanism for the discovery of new agents to fight antifungal drug resistance.

No abstract

This study aimed to characterize the antifungal effects of chiloscyphenol A (CA), a natural small molecule isolated from Chinese liverworts, and investigate its mode of action. CA was effective against five tested Candida species with a minimal inhibitory concentration (MIC) of 8–32 μg/ml and exhibited fungicidal activity against Candida albicans in both the planktonic state and mature biofilms. The in vivo study using Caenorhabditis elegans showed that CA prolonged the survival of C. albicans infected worms. Further investigations revealed that CA resulted in mitochondrial dysfunction as indicated by mtΔψ hyperpolarization, increased ATP production and intracellular ROS accumulation, and aggregated distribution of Tom70. In addition, CA caused perturbation of the cell membrane and increased membrane permeability, as demonstrated by specific staining and confocal microscopic and transmission electron microscopy (TEM) observations and by calcein-leakage measurements. This conclusion was further confirmed by the decreased cell size of CA-treated cells via three-dimensional contour-plot analysis using flow cytometry. Taken together, these results suggest that CA exerts fungicidal activity by eliciting both mitochondrial dysfunction and plasma membrane destruction in C. albicans. The elucidated mechanism supports the potential application of CA against clinical fungal infections.