β-Lapachone enhances the antifungal activity of fluconazole against a Pdr5p-mediated resistant Saccharomyces cerevisiae strain

Abstract

Objectives The aim of this study was to evaluate the ability of lapachones in disrupting the fungal multidrug resistance (MDR) phenotype, using a model of study which an azole-resistant Saccharomyces cerevisiae mutant strain that overexpresses the ATPbinding cassette (ABC) transporter Pdr5p. Methods The evaluation of the antifungal activity of lapachones and their possible synergism with fluconazole against the mutant S. cerevisiae strain was performed through broth microdilution and spot assays. Reactive oxygen species (ROS) and efflux pump activity were assessed by fluorometry. ATPase activity was evaluated by the Fiske and Subbarow method. The effect of βlapachone on PDR5 mRNA expression was assessed by RT-PCR. The release of hemoglobin was measured to evaluate the hemolytic activity of β-lapachone. Results α-nor-Lapachone and β-lapachone inhibited S. cerevisiae growth at 100 μg/ml. Only β-lapachone enhanced the antifungal activity of fluconazole, and this combined action was inhibited by ascorbic acid. β-Lapachone induced the production of ROS, inhibited Pdr5p-mediated efflux, and impaired Pdr5p ATPase activity. Also, β-lapachone neither affected the expression of PDR5 nor exerted hemolytic activity. Conclusions Data obtained indicate that β-lapachone is able to inhibit the S. cerevisiae efflux pump Pdr5p. Since this transporter is homologous to fungal ABC transporters, further studies employing clinical isolates that overexpress these proteins will be conducted to evaluate the effect of β-lapachone on pathogenic fungi.

Repurposing the FDA‐approved anticancer agent ponatinib as a fluconazole potentiator by suppression of multidrug efflux and Pma1 expression in a broad spectrum of yeast species

“…Pdr5 is an ABC transporter which pumps out a series of structurally unrelated compounds, including azoles and rhodamine (Prasad and Goffeau, 2012 ; Shekhar‐Guturja et al ., 2016b ; de Moraes et al ., 2020 ). Consistent with previous studies (Lamping et al ., 2007 ; Prasad and Goffeau, 2012 ), we also confirmed that fluconazole is a substrate of Pdr5.…”

Digoxin Derivatives Sensitize a Saccharomyces cerevisiae Mutant Strain to Fluconazole by Inhibiting Pdr5p

“…Oroidin, a sponge-derived alkaloid, inhibited R6G efflux by 60% at 200 μM [ 27 ]. Beta-lapachone, a naphthoquinone obtained from Tabebuia sp., inhibited Pdr5p activity by 79.4% at 100 μg/mL [ 28 ]. In this study, it was observed that digoxin derivatives inhibited R6G efflux by 76.65–100%.…”

Synthesis of Altissimacoumarin D and Other Prenylated Coumarins and Their Ability to Reverse the Multidrug Resistance Phenotype in Candida albicans

“…The toxicity of the coumarins was predicted in silico using OSIRIS Property Explorer [23], which provides information regarding mutagenic, tumorigenic, irritant, and reproductive effects, and GUSAR (General Unrestricted Structure-Activity Relationships), which provides a LD 50 (lethal dose to 50% of a given population) of the substances in rats, considering four routes of administration (intraperitoneal, intravenous, oral, and subcutaneous) [24].…”