Diazepam’s antifungal activity in fluconazole-resistant spp. and biofilm inhibition in : evaluation of the relationship with the proteins ALS3 and SAP5 Free

Abstract

The genus spp. has been highlighted as one of the main etiological agents causing fungal infections, with being the most prominent, responsible for most cases of candidemia. Due to its capacity for invasion and tissue adhesion, it is associated with the formation of biofilms, mainly in the environment and hospital devices, decreasing the effectiveness of available treatments. The repositioning of drugs, which is characterized by the use of drugs already on the market for other purposes, together with molecular-docking methods can be used aiming at the faster development of new antifungals to combat micro-organisms. This study aimed to evaluate the antifungal effect of diazepam on mature biofilms and its action on biofilm in formation, as well as its mechanism of action and interaction with structures related to the adhesion of , ALS3 and SAP5. To determine the MIC, the broth microdilution test was used according to protocol M27-A3 (CLSI, 2008). biofilm formation tests were performed using 96-well plates, followed by molecular-docking protocols to analyse the binding agent interaction with ALS3 and SAP5 targets. The results indicate that diazepam has antimicrobial activity against planktonic cells of spp. and biofilms, interacting with important virulence factors related to biofilm formation (ALS3 and SAP5). In addition, treatment with diazepam triggered a series of events in cells, such as loss of membrane integrity, mitochondrial depolarization and increased production of EROs, causing DNA damage and consequent cell apoptosis.

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2021-02-09
2024-03-29
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References

  1. Bongomin F, Gago S, Oladele R, Denning D. Global and multi-national prevalence of fungal diseases—estimate precision. J Fungi 2017; 3:57 [View Article]
    [Google Scholar]
  2. Chang YL, Yu S-J, Heitman J, Wellington M, Chen Y-L. New facets of antifungal therapy. Virulence 2017; 8:222–236 [View Article]
    [Google Scholar]
  3. Nicola AM, Albuquerque P, Paes HC, Fernandes L, Costa FF et al. Antifungal drugs: New insights in research & development. Pharmacol Ther 2019; 195:21–38 [View Article]
    [Google Scholar]
  4. Nucci M, Queiroz-Telles F, Alvarado-Matute T, Tiraboschi IN, Cortes J et al. Epidemiology of candidemia in Latin America: a laboratory-based survey. PLoS One 2013; 8:e59373 [View Article]
    [Google Scholar]
  5. Colombo AL, Júnior JNdeA, Guinea J. Emerging multidrug-resistant Candida species. Curr Opin Infect Dis 2017; 30:528–538 [View Article]
    [Google Scholar]
  6. Cesar J, França B, Elisa C, Ribeiro L. Candidemia in a Brazilian tertiary care hospital: incidence, frequency of different species, risk factors and antifungal susceptibility. Rev Soc Bras Med Trop 2008; 41:23–28
    [Google Scholar]
  7. Cavalheiro M, Teixeira MC. Candida biofilms: threats, challenges, and promising strategies. Front Med 2018; 5:1–15 [View Article]
    [Google Scholar]
  8. Reichhardt C, Stevens DA, Cegelski L. Fungal biofilm composition and opportunities in drug discovery. Future Med Chem 2016; 8:1455–1468 [View Article]
    [Google Scholar]
  9. Braga PR, Cruz IL, Ortiz I, Barreiros G, Nouér SA et al. Secular trends of candidemia at a Brazilian tertiary care teaching hospital. Brazilian J Infect Dis 2018; 22:273–277 [View Article]
    [Google Scholar]
  10. Brasil M da S. Ministário dA Saúde; 2017
  11. Doi AM, Pignatari ACC, Edmond MB, Marra AR, Camargo LFA et al. Epidemiology and microbiologic characterization of nosocomial candidemia from a Brazilian national surveillance program. PLoS One 2016; 11:e0146909 [View Article]
    [Google Scholar]
  12. Whaley SG, Berkow EL, Rybak JM, Nishimoto AT, Barker KS et al. Azole antifungal resistance in Candida albicans and emerging non-albicans Candida species. Front Microbiol 2017; 7:1–12 [View Article]
    [Google Scholar]
  13. Cho EJ, Shin JH, Kim SH, Kim HK, Park JS. Emergence of multiple resistance profiles involving azoles, echinocandins and amphotericin B in Candida glabrata isolates from a neutropenia patient with prolonged fungaemia. J Antimicrob Chemother 2014; 70:1268–1270
    [Google Scholar]
  14. Motta AL, de Almeida GMD, de Almeida Júnior JN, Burattini MN, Rossi F. Candidemia epidemiology and susceptibility profile in the largest Brazilian teaching hospital complex. Brazilian J Infect Dis 2010; 14:441–448 [View Article]
    [Google Scholar]
  15. Terças ALG, Marques SG, Moffa EB, Alves MB, de Azevedo CMPS et al. Antifungal drug susceptibility of Candida species isolated from HIV-positive patients recruited at a public hospital in São Luís, Maranhão, Brazil. Front Microbiol 2017; 8:1–8 [View Article]
    [Google Scholar]
  16. Kathwate GH, Shinde RB, Karuppayil SM. Antiepileptic drugs inhibit growth, dimorphism, and biofilm mode of growth in human pathogen Candida albicans . Assay Drug Dev Technol 2015; 13:307–312 [View Article]
    [Google Scholar]
  17. da Silva CR, de Andrade Neto JB, Sidrim JJC, Angelo MRF, Magalhães HIF et al. Synergistic effects of amiodarone and fluconazole on Candida tropicalis resistant to fluconazole. Antimicrob Agents Chemother 2013; 57:1691–1700 [View Article][PubMed]
    [Google Scholar]
  18. Razavi BM, Fazly Bazzaz BS, Sedigheh B, Bazzaz F. A review and new insights to antimicrobial action of local anesthetics. Eur J Clin Microbiol Infect Dis 2019; 38:991–1002 [View Article]
    [Google Scholar]
  19. Cui J, Ren B, Tong Y, Dai H, Zhang L. Synergistic combinations of antifungals and anti-virulence agents to fight against Candida albicans . Virulence 2015; 6:362–371 [View Article]
    [Google Scholar]
  20. Schleinkofer K, Wang T, Wade RC. Molecular docking. Encycl Ref Genomics Proteomics Mol Med 2006; 443:1149–1153
    [Google Scholar]
  21. Clinical Laboratory Standard Institute-CLSI-M27 A3 Reference method for broth dilution.
  22. Clinical Laboratory Standard Institute-CLSI CLSI M27-S4. Reference Method For Broth Dilution Antifungal Susceptibility Testing Of Yeasts; Fourth Informational Supplement 2012
    [Google Scholar]
  23. da Silva AR, de Andrade Neto JB, da Silva CR, Campos R de S, Costa Silva RA et al. Berberine antifungal activity in fluconazole-resistant pathogenic yeasts: action mechanism evaluated by flow cytometry and biofilm growth inhibition in Candida spp. Antimicrob Agents Chemother 2016; 60:3551–3557 [View Article]
    [Google Scholar]
  24. Neto JBA, Silva CR, Nascimento F, Sampaio LS, Silva AR. Screening of Antimicrobial Metabolite Yeast Isolates Derived Biome Ceará against Pathogenic Bacteria, Including MRSA : Antibacterial Activity and mode of Action Evaluated by Flow Cy. Original Research Article Screening of Antimicrobial Metabolite Yeas.
  25. Neto JBA, da Silva CR, Neta MAS, Campos RS, Siebra JT et al. Antifungal activity of naphthoquinoidal compounds in vitro against fluconazole-resistant strains of different Candida species: a special emphasis on mechanisms of action on Candida tropicalis . PLoS One 2014; 9:e93698–10 [View Article]
    [Google Scholar]
  26. de Andrade Neto JB, da Silva CR, Barroso FD, do Amaral Valente Sá LG, de Sousa Campos R et al. Synergistic effects of ketamine and azole derivatives on Candida spp. resistance to fluconazole. Future Microbiol 2020; 15:177–188 [View Article][PubMed]
    [Google Scholar]
  27. Pierce CG, Uppuluri P, Tristan AR, Wormley FL, Mowat E et al. A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc 2008; 3:1494–1500 [View Article]
    [Google Scholar]
  28. Costa Silva RA, da Silva CR, de Andrade Neto JB, da Silva AR, Campos RS et al. In vitro anti-Candida activity of selective serotonin reuptake inhibitors against fluconazole-resistant strains and their activity against biofilm-forming isolates. Microb Pathog 2017; 107:341–348 [View Article]
    [Google Scholar]
  29. Borelli C, Ruge E, Lee JH, Schaller M, Vogelsang A et al. X-ray structures of Sap1 and Sap5: structural comparison of the secreted aspartic proteinases from Candida albicans. Proteins 2008; 72:1308–1319 [View Article]
    [Google Scholar]
  30. Lin J, Oh S-H, Jones R, Garnett JA, Salgado PS et al. The peptide-binding cavity is essential for ALS3-mediated adhesion of Candida albicans to human cells. J Biol Chem 2014; 289:18401–18412 [View Article]
    [Google Scholar]
  31. Lucio FNM et al. Methylcytisine Alcaloid potentially active against dengue virus: a molecular docking study and electronic structural characterization; 2020
  32. Hanwell MD, Curtis DE, Lonie DC, Vandermeersch T, Zurek E. Avogadro: an advanced semantic chemical editor, visualization, and analysis platform; 2012
  33. Halgren TA. Merck molecular force field. 11; 2000; 17520–552
  34. Trott O, Olson AJ. Software news and update AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading; 2009
  35. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM. UCSF Chimera? A visualization system for exploratory research and analysis. J Comput Chem 2004; 25:1605–1612 [View Article]
    [Google Scholar]
  36. Dassault Systèmes BIOVIA Discovery Studio Visualizer, Versão 16.1.0.
  37. Yusuf D, Davis AM, Kleywegt GJ, Schmitt S. An Alternative Method for the Evaluation of Docking Performance: RSR vs RMSD; 20081411–1422
  38. Shityakov S, Foerster C. In silico predictive model to determine vector-mediated transport properties for the blood-brain barrier choline transporter. AABC 2014; 7:23–36 [View Article]
    [Google Scholar]
  39. Moraes DC, Ferreira-Pereira A. Insights on the anticandidal activity of non-antifungal drugs. J Mycol Med 2019; 29:253–259 [View Article]
    [Google Scholar]
  40. Chelli B, Lena A, Vanacore R, Da Pozzo E, Costa B et al. Peripheral benzodiazepine receptor ligands: mitochondrial transmembrane potential depolarization and apoptosis induction in rat C6 glioma cells. Biochem Pharmacol 2004; 68:125–134 [View Article][PubMed]
    [Google Scholar]
  41. Sarnowska A, Beręsewicz M, Zabłocka B, Domańska-Janik K. Diazepam neuroprotection in excitotoxic and oxidative stress involves a mitochondrial mechanism additional to the GABAAR and hypothermic effects. Neurochem Int 2009; 55:164–173 [View Article]
    [Google Scholar]
  42. Casellas P, Galiegue S, Basile AS. Peripheral benzodiazepine receptors and mitochondrial function. Neurochem Int 2002; 40:475–486 [View Article]
    [Google Scholar]
  43. Marino F, Cattaneo S, Cosentino M, Rasini E, Di Grazia L et al. Diazepam stimulates migration and phagocytosis of human neutrophils: possible contribution of peripheral-type benzodiazepine receptors and intracellular calcium. Pharmacology 2001; 63:42–49 [View Article]
    [Google Scholar]
  44. Tian H, Qu S, Wang Y, Lu Z, Zhang M et al. Calcium and oxidative stress mediate perillaldehyde-induced apoptosis in Candida albicans . Appl Microbiol Biotechnol 2017; 101:3335–3345 [View Article][PubMed]
    [Google Scholar]
  45. Azzopardi M, Farrugia G, Balzan R. Cell-cycle involvement in autophagy and apoptosis in yeast. Mech Ageing Dev 2017; 161:211–224 [View Article]
    [Google Scholar]
  46. Yun JE, Lee DG. Role of potassium channels in chlorogenic acid-induced apoptotic volume decrease and cell cycle arrest in Candida albicans . Biochim Biophys Acta - Gen Subj 1861; 2017:585–592
    [Google Scholar]
  47. Liu J-Y, Guo F, Wu H-L, Wang Y, Liu J-S. Midazolam anesthesia protects neuronal cells from oxidative stress-induced death via activation of the JNK-ERK pathway. Mol Med Rep 2017; 15:169–179 [View Article]
    [Google Scholar]
  48. Li B, Li X, Lin H, Zhou Y. Curcumin as a promising antibacterial agent: effects on metabolism and biofilm formation in S. mutans. Biomed Res Int 2018
    [Google Scholar]
  49. Madeo F, Fröhlich E, Ligr M, Grey M, Sigrist SJ et al. Oxygen stress: a regulator of apoptosis in yeast. J Cell Biol 1999; 145:757–767 [View Article]
    [Google Scholar]
  50. Simon H. Role of reactive oxygen species (ROS).pdf; 2000415–418
  51. Poliaková D, Sokolı Ková B, Kolarov J, Šabova L'udmila. The antiapoptotic protein Bcl-x(L) prevents the cytotoxic effect of Bax, but not Bax-induced formation of reactive oxygen species, in Kluyveromyces lactis. Microbiology 2002; 148:2789–2795 [View Article][PubMed]
    [Google Scholar]
  52. Cheng J, Park T-S, Chio L-C, Fischl AS, Ye XS. Induction of apoptosis by sphingoid long-chain bases in Aspergillus nidulans. Mol Cell Biol 2003; 23:163–177 [View Article][PubMed]
    [Google Scholar]
  53. Balzan R, Sapienza K, Galea DR, Vassallo N, Frey H et al. Aspirin commits yeast cells to apoptosis depending on carbon source. Microbiology 2004; 150:109–115 [View Article]
    [Google Scholar]
  54. Trancíková A, Weisová P, Kiššová I, Zeman I, Kolarov J. Production of reactive oxygen species and loss of viability in yeast mitochondrial mutants: protective effect of Bcl-x. FEMS Yeast Res 2004; 5:149–156 [View Article]
    [Google Scholar]
  55. Kim S, Hwang JS, Lee DG. Lactoferricin B like peptide triggers mitochondrial disruption-mediated apoptosis by inhibiting respiration under nitric oxide accumulation in Candida albicans . IUBMB Life 2020; 72:1515–1527 [View Article][PubMed]
    [Google Scholar]
  56. Carmona-Gutierrez D, Bauer MA, Zimmermann A, Aguilera A, Austriaco N et al. Guidelines and recommendations on yeast cell death nomenclature. Microb Cell 2018; 5:4–31 [View Article]
    [Google Scholar]
  57. Ravagnan L, Roumier T, Kroemer G. Mitochondria, the killer organelles and their weapons. J Cell Physiol 2002; 192:131–137 [View Article]
    [Google Scholar]
  58. Wang T, Shao J, Da W, Li Q, Shi G et al. Strong synergism of palmatine and Fluconazole/Itraconazole against planktonic and biofilm cells of Candida species and Efflux-associated antifungal mechanism. Front Microbiol 2018; 9:1–12 [View Article]
    [Google Scholar]
  59. Martin SJ, Reutelingsperger CP, McGahon AJ, Rader JA, van Schie RC et al. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med 1995; 182:1545–1556 [View Article][PubMed]
    [Google Scholar]
  60. Filler SG. Candida–host cell receptor–ligand interactions. Curr Opin Microbiol 2006; 9:333–339 [View Article]
    [Google Scholar]
  61. Kwan P, Sills GJ, Brodie MJ. The mechanisms of action of commonly used antiepileptic drugs. Pharmacol Ther 2001; 90:21–34 [View Article]
    [Google Scholar]
  62. Tobudic S, Kratzer C, Lassnigg A, Presterl E. Antifungal susceptibility of Candida albicans in biofilms. Mycoses 2012; 55:199–204 [View Article]
    [Google Scholar]
  63. de Barros PP, Rossoni RD, De Camargo Ribeiro F, Junqueira JC, Jorge AOC. Temporal profile of biofilm formation, gene expression and virulence analysis in Candida albicans strains. Mycopathologia 2017; 182:285–295 [View Article][PubMed]
    [Google Scholar]
  64. Fan Y, He H, Dong Y, Pan H. Hyphae-specific genes HGC1, ALS3, Hwp1, and ECE1 and relevant signaling pathways in Candida albicans . Mycopathologia 2013; 176:329–335 [View Article][PubMed]
    [Google Scholar]
  65. Kullberg BJ, Vasquez J, Mootsikapun P, Nucci M, Paiva JA et al. Efficacy of anidulafungin in 539 patients with invasive candidiasis: a patient-level pooled analysis of six clinical trials. J Antimicrob Chemother 2017; 72:2368–2377 [View Article]
    [Google Scholar]
  66. Mushi MF, Bader O, Taverne-Ghadwal L, Bii C, Groß U et al. Oral candidiasis among African human immunodeficiency virus-infected individuals: 10 years of systematic review and meta-analysis from sub-Saharan Africa. J Oral Microbiol 2017; 9:1317579 [View Article]
    [Google Scholar]
  67. Fu Y, Phan QT, Luo G, Solis NV, Liu Y et al. Investigation of the function of Candida albicans Als3 by heterologous expression in Candida glabrata. Infect Immun 2013; 81:2528–2535 [View Article]
    [Google Scholar]
  68. Green CB, Zhao X, Hoyer LL. Use of green fluorescent protein and reverse transcription-PCR to monitor Candida albicans agglutinin-like sequence gene expression in a murine model of disseminated candidiasis. Infect Immun 2005; 73:1852–1855 [View Article]
    [Google Scholar]
  69. Joo MY, Shin JH, Jang H-C, Song ES, Kee SJ et al. Expression of SAP5 and SAP9 in Candida albicans biofilms: comparison of bloodstream isolates with isolates from other sources. Med Mycol 2013; 51:892–896 [View Article]
    [Google Scholar]
  70. Winter MB, Salcedo EC, Lohse MB, Hartooni N, Gulati M et al. Global identification of biofilm-specific proteolysis in Candida albicans . MBio 2016; 7:1–13 [View Article]
    [Google Scholar]
  71. Jatlow P, Dobular K, Bailey D. Serum diazepam concentrations in overdose. Their significance. Am J Clin Pathol 1979; 72:571–577
    [Google Scholar]
  72. Fadul JA. Encyclopedia of Theory & Practice in Psychotherapy & Counseling, 1st ed. Raleigh: Lulu Press Inc; 2015
    [Google Scholar]
  73. Hang MTH, Smith BE, Keck C, Keshavarzian A, Sedghi S. Increasing efficacy and reducing side effects in treatment of chronic anal fissures: a study of topical diazepam therapy. Medicine 2017; 96:e6853
    [Google Scholar]
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