Skip to main content

Advertisement

SpringerLink
Log in

Inhibition of ergosterol synthesis in Candida albicans by novel eugenol tosylate congeners targeting sterol 14α-demethylase (CYP51) enzyme

  • Original Paper
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

This study is a continuation and extension of our previous study in which we synthesized seven novel eugenol tosylate congeners (ETC-1 to ETC-7) from a natural compound eugenol and checked their antifungal activity against different isolates of Candida albicans. All these ETCs showed potent antifungal activity to varying degrees. In this study, the aim is to evaluate the effect of most active compounds (ETC-5, ETC-6 and ETC-7) on ergosterol biosynthesis pathway and cellular viability in C. albicans by applying combined approach of in silico and in vitro methodologies. In silico studies were done through all atom molecular mechanics approach and free binding energy estimations, and in vitro study was done by estimating total intracellular sterol content and effect on expression of ERG11 gene. Furthermore, effect on cell viability by these compounds was also tested. Our results demonstrated that these ETCs target ergosterol biosynthesis pathway in C. albicans by inhibiting the lanosterol 14-α demethylase enzyme and also downregulates expression of its related gene ERG11. Furthermore, these ETCs exhibit potent fungicidal effect in cell viability assay, thus overall results advocating the claim that these tosylates have potential to be taken to next level of antifungal drug development.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Data availability

The data that support the findings of this study are available from the corresponding author upon request.

References

  • Ahmad A, Khan A, Manzoor N, Khan LA (2010) Evolution of ergosterol biosynthesis inhibitors as fungicidal against Candida. Microb Pathogen 48(1):35–41

    Article  CAS  Google Scholar 

  • Ahmad A, Khan A, Akhtar F, Yousuf S, Xess I, Khan LA, Manzoor N (2011) Fungicidal activity of thymol and carvacrol by disrupting ergosterol biosynthesis and membrane integrity against Candida. Eur J Clin Microbiol Infect Dis 30(1):41–50

    Article  CAS  PubMed  Google Scholar 

  • Ahmad A, Wani MY, Khan A, Manzoor N, Molepo J (2015) Synergistic interactions of Eugenol-tosylate and its congeners with fluconazole against Candida albicans. PLoS ONE 10:e0145053

    Article  PubMed  PubMed Central  Google Scholar 

  • Alizadeh F, Khodavandi A, Zalakian S (2017) Quantitation of ergosterol content and gene expression profile of ERG11 gene in fluconazole-resistant Candida albicans. Curr Med Mycol 3(1):13–19

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Alvarez-Rueda N, Fleury A, Logé C, Pagniez F, Robert E, Morio F (2016) The amino acid substitution N136Y in Candida albicans sterol 14alpha-demethylase is involved in fluconazole resistance. Med Mycol 54(7):764–775

    Article  CAS  PubMed  Google Scholar 

  • Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE (2000) The protein data bank. Nucleic Acids Res 28(1):235–242

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brown GD, Denning DW, Gow NA, Levitz SM, Netea MG, White TC (2012) Hidden killers: human fungal infections. Sci Transl Med 4:165rv13

    Article  PubMed  Google Scholar 

  • Carrasco H, Raimondi M, Svetaz L, Liberto M, Rodriguez MV, Espinoza L, Madrid A, Zacchino S (2012) Antifungal activity of eugenol analogues. Influence of different substituents and studies on mechanism of action. Molecules 17(1):1002–1024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • de Oliveira CG, Vilas-Boas LA, Castilho MS, Carazzolle MF, Pirovani CP, Selbach-Schnadelbach A, Gramacho KP, Ramos PI, Barbosa LV, Pereira GA, Góes-Neto A (2014) Analysis of the ergosterol biosynthesis pathway cloning, molecular characterization and phylogeny of lanosterol 14 α-demethylase (ERG11) gene of Moniliophthora perniciosa. Genet Mol Biol 37(4):683–693

    Article  Google Scholar 

  • Dias DA, Urban S, Roessner U (2012) A historical overview of natural products in drug discovery. Metabolites 2(2):303–336

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eddouzi J, Parker JE, Vale-Silva LA, Coste A, Ischer F, Kelly S, Manai M, Sanglard D (2013) Molecular mechanisms of drug resistance in clinical Candida species isolated from Tunisian hospitals. Antimicrob Agents Chemother 57(7):3182–3193

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Genheden S, Ryde U (2015) The MM/PBSA and MM/GBSA methods to estimate ligand-binding affinities. Expert Opin Drug Discov 10(5):449–461

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Harvey MJ, De Fabritiis G (2009) An implementation of the smooth particle Mesh Ewald Method on GPU hardware. J Chem Theory Comput 5(9):2371–2377

    Article  CAS  PubMed  Google Scholar 

  • Hata M, Ishii Y, Watanabe E, Uoto K, Kobayashi S, Yoshida K, Otani T, Ando A (2010) Inhibition of ergosterol synthesis by novel antifungal compounds targeting C-14 reductase. Med Mycol 48(4):613–621

    Article  CAS  PubMed  Google Scholar 

  • Hipólito TMM, Bastos GTL, Barbosa TWL, de Souza TB, Coelho LFL, Dias ALT, Rodríguez IC, Dos Santos MH, Dias DF, Franco LL, Carvalho DT (2018) Synthesis, activity, and docking studies of eugenol-based glucosides as new agents against Candida sp. Chem Biol Drug Des 92(2):1514–1524

    Article  PubMed  Google Scholar 

  • Keniya MV, Sabherwal M, Wilson RK, Woods MA, Sagatova AA, Tyndall JDA, Monk BC (2018) Crystal structures of full-length lanosterol 14α-demethylases of prominent fungal pathogens Candida albicans and Candida glabrata provide tools for antifungal discovery. Antimicrob Agents Chemother 62(11):e01134

    PubMed  PubMed Central  Google Scholar 

  • Lindberg E, Hammarström H, Ataollahy N, Kondori N (2019) Species distribution and antifungal drug susceptibilities of yeasts isolated from the blood samples of patients with candidemia. Sci Rep 9(1):3838

    Article  PubMed  PubMed Central  Google Scholar 

  • Liu X, Ma Z, Zhang J, Yang L (2017) Antifungal compounds against Candida infections from traditional Chinese medicine. Biomed Res Int 2017:4614183

    PubMed  PubMed Central  Google Scholar 

  • Mushtaq S, Abbasi BH, Uzair B, Abbasi R (2018) Natural products as reservoirs of novel therapeutic agents. EXCLI J 17:420–451

    PubMed  PubMed Central  Google Scholar 

  • Nakayama H, Nakayama N, Arisawa M, Aoki Y (2001) In vitro and in vivo effects of 14alpha-demethylase (ERG11) depletion in Candida glabrata. Antimicrob Agents Chemother 45(11):3037–3045

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pan SY, Zhou SF, Gao SH, Yu ZL, Zhang SF, Tang MK, Sun JN, Ma DL, Han YF, Fong WF, Ko KM (2013) New perspectives on how to discover drugs from herbal medicines: Cam's outstanding contribution to modern therapeutics. Evid Based Complement Alternat Med 2013:627375

    PubMed  PubMed Central  Google Scholar 

  • Perez A, MacCallum JL, Brini E, Simmerling C, Dill KA (2015) Grid-based backbone correction to the ff12SB protein force field for implicit-solvent simulations. J Chem Theory Comput 11(10):4770–4779

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE (2004) UCSF Chimera—a visualization system for exploratory research and analysis. J Comput Chem 25(13):1605–1612

    Article  CAS  PubMed  Google Scholar 

  • Piccione D, Mirabelli S, Minto N, Bouklas T (2019) Difficult but not impossible: in search of an anti-candida vaccine. Curr Trop Med Rep 6:42–49

    Article  Google Scholar 

  • Rajput SB, Karuppayil SM (2013) Small molecules inhibit growth, viability and ergosterol biosynthesis in Candida albicans. Springerplus 2(1):26

    Article  PubMed  PubMed Central  Google Scholar 

  • Roe DR, Cheatham TE (2013) PTRAJ and CPPTRAJ: software for processing and analysis of molecular dynamics trajectory data. J Chem Theory Comput 9(7):3084–3095

    Article  CAS  PubMed  Google Scholar 

  • Sanglard D (2016) Emerging threats in antifungal-resistant fungal pathogens. Front Med (Lausanne) 3:11

    Google Scholar 

  • Sanglard D, Ischer F, Parkinson T, Falconer D, Bille J (2003) Candida albicans mutations in the ergosterol biosynthetic pathway and resistance to several antifungal agents. Antimicrob Agents Chemother 47(8):2404–2412

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sastry GM, Adzhigirey M, Day T, Annabhimoju R, Sherman W (2013) Protein and ligand preparation: parameters, protocols, and influence on virtual screening enrichments. J Comput Aided Mol Des 27(3):221–234

    Article  PubMed  Google Scholar 

  • Sievert DM, Ricks P, Edwards JR, Schneider A, Patel J, Srinivasan A, Kallen A, Limbago B, Fridkin S, National Healthcare Safety Network (NHSN) Team, and Participating NHSN Facilities (2013) Antimicrobial-resistant pathogens associated with healthcare-associated infections: summary of data reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2009–2010. Infect Control Hosp Epidemiol 34(1):1–14

    Article  PubMed  Google Scholar 

  • Silva FFM, Monte FJQ, Lemos TLG, Nascimento PGG, Costa AKM, Paiva LMM (2018) Eugenol derivatives: synthesis, characterization, and evaluation of antibacterial and antioxidant activities. Chem Cent J 12(1):34

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Trott O, Olson AJ (2010) AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem 31(2):455–461

    CAS  PubMed  PubMed Central  Google Scholar 

  • van der Spoel D, van Maaren PJ (2006) The origin of layer structure artifacts in simulations of liquid water. J Chem Theory Comput 2(1):1–11

    Article  PubMed  Google Scholar 

  • Xu Y, Sheng F, Zhao J, Chen L, Li C (2015) ERG11 mutations and expression of resistance genes in fluconazole-resistant Candida albicans isolates. Arch Microbiol 197(9):1087–1093

    Article  CAS  PubMed  Google Scholar 

  • Yu LH, Wei X, Ma M, Chen XJ, Xu SB (2012) Possible inhibitory molecular mechanism of farnesol on the development of fluconazole resistance in Candida albicans biofilm. Antimicrob Agents Chemother 56(2):770–775

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

We gratefully acknowledge financial support from University Research Committee Grant for 2019—Friedel Sellschop Award (Grant no: AZMD019), Wits Faculty of Health Sciences Research Committee (FRC, Grant no: 001…5254) and South Africa Medical Research Council (MRC) (Grant no: MLEP016).

Author information

Authors and Affiliations

  1. Clinical Microbiology and Infectious Diseases, School of Pathology, Health Sciences, University of the Witwatersrand, Johannesburg, 2193, South Africa

    Shabir Ahmad Lone, Shama Khan & Aijaz Ahmad

  2. Infection Control, Charlotte Maxeke Johannesburg Academic Hospital, National Health Laboratory Service, Johannesburg, 2193, South Africa

    Aijaz Ahmad

Authors
  1. Shabir Ahmad Lone
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shama Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Aijaz Ahmad
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceived and designed the experiments: AASK. Performed the experiments: SAL. Analyzed the data: SAL, SK, AA. Contributed reagents/materials/analysis tools: AA. Wrote the paper: SAL.

Corresponding author

Correspondence to Aijaz Ahmad.

Ethics declarations

Conflict of interest

We have no competing interests to declare.

Ethical approval

This study was approved by the Human Research Ethics Committee of University of the Witwatersrand (Johannesburg, South Africa). Existing stock cultures of C. albicans used in this study were stored in the Department of Clinical Microbiology and Infectious Diseases, University of the Witwatersrand, Johannesburg, South Africa.

Additional information

Communicated by Erko Stackebrandt.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 134 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lone, S.A., Khan, S. & Ahmad, A. Inhibition of ergosterol synthesis in Candida albicans by novel eugenol tosylate congeners targeting sterol 14α-demethylase (CYP51) enzyme. Arch Microbiol 202, 711–726 (2020). https://doi.org/10.1007/s00203-019-01781-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00203-019-01781-2

Keywords

Search

Navigation