β-Lapachone Enhances the Antifungal Activity of Fluconazole Against a Pdr5p-mediated Resistant Saccharomyces Cerevisiae Strain

Overview

Journal Braz J Microbiol

Specialty Microbiology

Date 2020 Mar 12

PMID 32157667

Citations 5

Authors

Daniel Clemente de Moraes

Karina Martins Cardoso

Levy Tenorio Sousa Domingos

Maria do Carmo Freire Ribeiro Pinto

Robson Q Monteiro

Antonio Ferreira-Pereira

Affiliations

Soon will be listed here.

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 ATP-binding 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 β-lapachone 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.

Citing Articles

Batzelladine D, a Marine Natural Product, Reverses the Fluconazole Resistance Phenotype Mediated by Transmembrane Transporters in and Interferes with Its Biofilm: An In Vitro and In Silico Study.

Domingos L, de Moraes D, Santos M, Curvelo J, Bayona-Pacheco B, Marquez E Mar Drugs. 2024; 22(11).

PMID: 39590782 PMC: 11595962. DOI: 10.3390/md22110502.

Antifungal activity of β-lapachone against a fluconazole-resistant Candida auris strain.

de Moraes D, Rollin-Pinheiro R, Pinto M, Domingos L, Barreto-Bergter E, Ferreira-Pereira A Braz J Microbiol. 2024; 55(3):2593-2601.

PMID: 38743245 PMC: 11405563. DOI: 10.1007/s42770-024-01375-1.

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

Silva A, de Moraes D, do Carmo D, Gomes G, Ganesan A, Lopes R J Fungi (Basel). 2023; 9(7).

PMID: 37504746 PMC: 10381857. DOI: 10.3390/jof9070758.

Digoxin Derivatives Sensitize a Mutant Strain to Fluconazole by Inhibiting Pdr5p.

de Moraes D, Tessis A, Rollin-Pinheiro R, Princival J, Villar J, Barbosa L J Fungi (Basel). 2022; 8(8).

PMID: 35893137 PMC: 9330353. DOI: 10.3390/jof8080769.

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.

Liu L, Jiang T, Zhou J, Mei Y, Li J, Tan J Microb Biotechnol. 2021; 15(2):482-498.

PMID: 33955652 PMC: 8867973. DOI: 10.1111/1751-7915.13814.

References

Keniya M, Fleischer E, Klinger A, Cannon R, Monk B . Inhibitors of the Candida albicans Major Facilitator Superfamily Transporter Mdr1p Responsible for Fluconazole Resistance. PLoS One. 2015; 10(5):e0126350. PMC: 4423874. DOI: 10.1371/journal.pone.0126350. View

Dudoignon E, Alanio A, Anstey J, Depret F, Coutrot M, Fratani A . Outcome and potentially modifiable risk factors for candidemia in critically ill burns patients: A matched cohort study. Mycoses. 2018; 62(3):237-246. DOI: 10.1111/myc.12872. View

Rosseti I, Rocha J, Costa M . Diphenyl diselenide (PhSe)2 inhibits biofilm formation by Candida albicans, increasing both ROS production and membrane permeability. J Trace Elem Med Biol. 2014; 29:289-95. DOI: 10.1016/j.jtemb.2014.08.001. View

Prasad R, Rawal M . Efflux pump proteins in antifungal resistance. Front Pharmacol. 2014; 5:202. PMC: 4148622. DOI: 10.3389/fphar.2014.00202. View

Niimi K, Harding D, Parshot R, King A, Lun D, Decottignies A . Chemosensitization of fluconazole resistance in Saccharomyces cerevisiae and pathogenic fungi by a D-octapeptide derivative. Antimicrob Agents Chemother. 2004; 48(4):1256-71. PMC: 375246. DOI: 10.1128/AAC.48.4.1256-1271.2004. View

Chazotte B . Labeling mitochondria with JC-1. Cold Spring Harb Protoc. 2011; 2011(9). DOI: 10.1101/pdb.prot065490. View

Decottignies A, Kolaczkowski M, Balzi E, Goffeau A . Solubilization and characterization of the overexpressed PDR5 multidrug resistance nucleotide triphosphatase of yeast. J Biol Chem. 1994; 269(17):12797-803. View

Yang Y, Zhou X, Xu M, Piao J, Zhang Y, Lin Z . β-lapachone suppresses tumour progression by inhibiting epithelial-to-mesenchymal transition in NQO1-positive breast cancers. Sci Rep. 2017; 7(1):2681. PMC: 5457413. DOI: 10.1038/s41598-017-02937-0. View

Spitzer M, Robbins N, Wright G . Combinatorial strategies for combating invasive fungal infections. Virulence. 2016; 8(2):169-185. PMC: 5354157. DOI: 10.1080/21505594.2016.1196300. View

10.

Cavalheiro M, Pais P, Galocha M, Teixeira M . Host-Pathogen Interactions Mediated by MDR Transporters in Fungi: As Pleiotropic as it Gets!. Genes (Basel). 2018; 9(7). PMC: 6071111. DOI: 10.3390/genes9070332. View

11.

Anaissi-Afonso L, Oramas-Royo S, Ayra-Plasencia J, Martin-Rodriguez P, Garcia-Luis J, Lorenzo-Castrillejo I . Lawsone, Juglone, and β-Lapachone Derivatives with Enhanced Mitochondrial-Based Toxicity. ACS Chem Biol. 2018; 13(8):1950-1957. DOI: 10.1021/acschembio.8b00306. View

12.

Menacho-Marquez M, Murguia J . Beta-lapachone activates a Mre11p-Tel1p G1/S checkpoint in budding yeast. Cell Cycle. 2006; 5(21):2509-16. DOI: 10.4161/cc.5.21.3394. View

13.

Hwang J, Choi H, Kim A, Yun J, Yu R, Woo E . Hibicuslide C-induced cell death in Candida albicans involves apoptosis mechanism. J Appl Microbiol. 2014; 117(5):1400-11. DOI: 10.1111/jam.12633. View

14.

Menacho-Marquez M, Rodriguez-Hernandez C, Villaronga M, Perez-Valle J, Gadea J, Belandia B . eIF2 kinases mediate β-lapachone toxicity in yeast and human cancer cells. Cell Cycle. 2015; 14(4):630-40. PMC: 5154358. DOI: 10.4161/15384101.2014.994904. View

15.

Ivnitski-Steele I, Holmes A, Lamping E, Monk B, Cannon R, Sklar L . Identification of Nile red as a fluorescent substrate of the Candida albicans ATP-binding cassette transporters Cdr1p and Cdr2p and the major facilitator superfamily transporter Mdr1p. Anal Biochem. 2009; 394(1):87-91. PMC: 2739806. DOI: 10.1016/j.ab.2009.07.001. View

16.

Wang C, Zhang Y, Zhang W, Yuan S, Ng T, Ye X . Purification of an Antifungal Peptide from Seeds of var. and Investigation of Its Antifungal Activity and Mechanism of Action. Molecules. 2019; 24(7). PMC: 6480268. DOI: 10.3390/molecules24071337. View

17.

Loo T, Clarke D . Tariquidar inhibits P-glycoprotein drug efflux but activates ATPase activity by blocking transition to an open conformation. Biochem Pharmacol. 2014; 92(4):558-66. DOI: 10.1016/j.bcp.2014.10.006. View

18.

Pereira Rangel L, Fritzen M, Yunes R, Leal P, Creczynski-Pasa T, Ferreira-Pereira A . Inhibitory effects of gallic acid ester derivatives on Saccharomyces cerevisiae multidrug resistance protein Pdr5p. FEMS Yeast Res. 2010; 10(3):244-51. DOI: 10.1111/j.1567-1364.2010.00603.x. View

19.

Ricardo E, Costa-de-Oliveira S, Dias A, Guerra J, Rodrigues A, Pina-Vaz C . Ibuprofen reverts antifungal resistance on Candida albicans showing overexpression of CDR genes. FEMS Yeast Res. 2009; 9(4):618-25. DOI: 10.1111/j.1567-1364.2009.00504.x. View

20.

Ramos-Perez C, Lorenzo-Castrillejo I, Quevedo O, Garcia-Luis J, Matos-Perdomo E, Medina-Coello C . Yeast cytotoxic sensitivity to the antitumour agent β-lapachone depends mainly on oxidative stress and is largely independent of microtubule- or topoisomerase-mediated DNA damage. Biochem Pharmacol. 2014; 92(2):206-19. DOI: 10.1016/j.bcp.2014.09.006. View