Bis-guanylhydrazones as efficient anti-Candida compounds through DNA interaction
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Candida spp. are leading causes of opportunistic mycoses, including life-threatening hospital-borne infections, and novel antifungals, preferably aiming targets that have not been used before, are constantly needed. Hydrazone- and guanidine-containing molecules have shown a wide range of biological activities, including recently described excellent antifungal properties. In this study, four bis-guanylhydrazone derivatives (BG1–4) were generated following a previously developed synthetic route. Anti-Candida (two C. albicans, C. glabrata, and C. parapsilosis) minimal inhibitory concentrations (MICs) of bis-guanylhydrazones were between 2 and 15.6 μg/mL. They were also effective against preformed 48-h-old C. albicans biofilms. In vitro DNA interaction, circular dichroism, and molecular docking analysis showed the great ability of these compounds to bind fungal DNA. Competition with DNA-binding stain, exposure of phosphatidylserine at the outer layer of the cytoplasmic membrane, and activation of metacaspases were shown for BG3. This pro-apoptotic effect of BG3 was only partially due to the accumulation of reactive oxygen species in C. albicans, as only twofold MIC and higher concentrations of BG3 caused depolarization of mitochondrial membrane which was accompanied by the decrease of the activity of fungal mitochondrial dehydrogenases, while the activity of oxidative stress response enzymes glutathione reductase and catalase was not significantly affected. BG3 showed synergistic activity with amphotericin B with a fractional inhibitory concentration index of 0.5. It also exerted low cytotoxicity and the ability to inhibit epithelial cell (TR146) invasion and damage by virulent C. albicans SC5314. With further developments, BG3 may further progress in the antifungal pipeline as a DNA-targeting agent.
KeywordsAntifungal activity Candida spp. Bis-guanylhydrazone DNA interaction ROS generation Synergy
This work was supported by the Ministry of Education, Science and Technological Development of Serbia (Grant Nos. 172008 and 173048). Research Grant 2015 by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) to JNR is also acknowledged. Work performed in Jena was funded by the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie Grant Agreement number 642095 (OPATHY) to MP.
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Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Ajdačić V, Senerovic L, Vranić M, Pekmezovic M, Arsic-Arsenijevic V, Veselinovic A, Veselinovic J, Šolaja BA, Nikodinovic-Runic J, Opsenica IM (2016) Synthesis and evaluation of thiophene-based guanylhydrazones (iminoguanidines) efficient against panel of voriconazole-resistant fungal isolates. Bioorg Med Chem 24(6):1277–1291. https://doi.org/10.1016/j.bmc.2016.01.058 CrossRefPubMedGoogle Scholar
- Beccia MR, Biver T, Pardini A, Spinelli J, Secco F, Venturini M, Busto Vazquez N, Lopez Cornejo MP, Martin Herrera VI, Prado Gotor R (2012) The fluorophore 4′,6-diamidino-2-phenylindole (DAPI) induces DNA folding in long double-stranded DNA. Chem Asian J 7(8):1803–1810. https://doi.org/10.1002/asia.201200177 CrossRefPubMedGoogle Scholar
- Clinical and Laboratory Standards Institute (2008) Reference method for broth dilution antifungal susceptibility testing of yeasts—Third Edition: Approved Standard M27-A3. CLSI W, PA, USAGoogle Scholar
- Clinical and Laboratory Standards Institute (2012) Reference method for broth dilution antifungal susceptibility testing of yeasts: fourth informational supplement M27-S4. CLSI W, PA, USAGoogle Scholar
- Clinical and Laboratory Standards Institute (2015) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; Approved Standard—Tenth Edition M07-A10. CLSI W, PA, USAGoogle Scholar
- Denning DW, Perlin DS, Muldoon EG, Colombo AL, Chakrabarti A, Richardson MD, Sorrell TC (2017) Delivering on antimicrobial resistance agenda not possible without improving fungal diagnostic capabilities. Emerg Infect Dis 23(2):177–183. https://doi.org/10.3201/eid2302.152042 CrossRefPubMedPubMedCentralGoogle Scholar
- Godoy JSR, Kioshima ÉS, Abadio AKR, Felipe MSS, de Freitas SM, Svidzinski TIE (2016) Structural and functional characterization of the recombinant thioredoxin reductase from Candida albicans as a potential target for vaccine and drug design. Appl Microbiol Biotechnol 100(9):4015–4025. https://doi.org/10.1007/s00253-015-7223-8 CrossRefPubMedGoogle Scholar
- Gowda KRS, Mathew BB, Sudhamani CN, Naik HSB (2014) Mechanism of DNA binding and cleavage. Biomed Biotechnol 2:1–9Google Scholar
- Maiolo EM, Furustrand Tafin U, Borens O, Trampuz A (2014) Activities of fluconazole, caspofungin, anidulafungin, and amphotericin B on planktonic and biofilm Candida species determined by microcalorimetry. Antimicrob Agents Chemother 58(5):2709–2717. https://doi.org/10.1128/AAC.00057-14 CrossRefPubMedPubMedCentralGoogle Scholar
- Moyes DL, Wilson D, Richardson JP, Mogavero S, Tang SX, Wernecke J, Höfs S, Gratacap RL, Robbins J, Runglall M, Murciano C, Blagojevic M, Thavaraj S, Förster TM, Hebecker B, Kasper L, Vizcay G, Iancu SI, Kichik N, Häder A, Kurzai O, Luo T, Krüger T, Kniemeyer O, Cota E, Bader O, Wheeler RT, Gutsmann T, Hube B, Naglik JR (2016) Candidalysin is a fungal peptide toxin critical for mucosal infection. Candidalysin is a fungal peptide toxin critical for mucosal infection. Nature 532(7597):64–68. https://doi.org/10.1038/nature17625 CrossRefPubMedPubMedCentralGoogle Scholar
- Pierce CG, Uppuluri P, Tristan AR, Wormley FL Jr, Mowat E, Ramage G, Lopez-Ribot JL (2008) A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc 3(9):1494–1500. https://doi.org/10.1038/nprot.2008.141 CrossRefPubMedPubMedCentralGoogle Scholar
- Shrestha SK, Kril LM, Green KD, Kwiatkowski S, Sviripa VM, Nickell JR, Dwoskin LP, Watt DS, Garneau-Tsodikova S (2017) Bis(N-amidinohydrazones) and N-(amidino)-N′-aryl-bishydrazones: new classes of antibacterial/antifungal agents. Bioorg Med Chem 25(1):58–66. https://doi.org/10.1016/j.bmc.2016.10.009 CrossRefPubMedGoogle Scholar
- Silva S, Negri M, Henriques M, Oliveira R, Williams DW, Azeredo J (2012) Candida glabrata, Candida parapsilosis and Candida tropicalis: biology, epidemiology, pathogenicity and antifungal resistance. FEMS Microbiol Rev 36(2):288–305. https://doi.org/10.1111/j.1574-6976.2011.00278.x CrossRefPubMedGoogle Scholar
- Thewes S, Moran GP, Magee BB, Schaller M, Sullivan DJ, Hube B (2008) Phenotypic screening, transcriptional profiling, and comparative genomic analysis of an invasive and non-invasive strain of Candida albicans. BMC Microbiol 8(1):187. https://doi.org/10.1186/1471-2180-8-187 CrossRefPubMedPubMedCentralGoogle Scholar
- Uppuluri P, Srinivasan A, Ramasubramanian A, Lopez-Ribot J (2011) Effects of fluconazole, amphotericin B, and Caspofungin on Candida albicans biofilms under conditions of flow and on biofilm dispersion. Antimicrob Agents Chemother 55(7):3591–3593. https://doi.org/10.1128/AAC.01701-10 CrossRefPubMedPubMedCentralGoogle Scholar
- Wachtler B, Wilson D, Haedicke K, Dalle F, Hube B (2011) From attachment to damage: defined genes of Candida albicans mediate adhesion, invasion and damage during interaction with oral epithelial cells. PLoS One 6(2):e17046. https://doi.org/10.1371/journal.pone.0017046 CrossRefPubMedPubMedCentralGoogle Scholar
- Wachtler B, Citiulo F, Jablonowski N, Forster S, Dalle F, Schaller M, Wilson D, Hube B (2012) Candida albicans-epithelial interactions: dissecting the roles of active penetration, induced endocytosis and host factors on the infection process. PLoS One 7(5):e36952. https://doi.org/10.1371/journal.pone.0036952 CrossRefPubMedPubMedCentralGoogle Scholar
- Wu XZ, Chang WQ, Cheng AX, Sun LM, Lou HX (2010) Plagiochin E, an antifungal active macrocyclic bis(bibenzyl), induced apoptosis in Candida albicans through a metacaspase-dependent apoptotic pathway. Biochim Biophys Acta 1800(4):439–347. https://doi.org/10.1016/j.bbagen.2010.01.001 CrossRefPubMedGoogle Scholar