Skip to main content

Advertisement

SpringerLink
Log in

Structure-based design of δ-lactones for new antifungal drug development: susceptibility, mechanism of action, and toxicity

  • Original Article
  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Dermatophytes are the etiological agents of cutaneous mycoses, including the prevalent nail infections and athlete’s foot. Candida spp. are opportunistic and emerging pathogens, causing superficial to deeper infections related to high mortality rates. As a consequence of prolonged application of antifungal drugs, the treatment failures combined with multidrug-resistance have become a serious problem in clinical practice. Therefore, novel alternative antifungals are required urgently. δ-Lactones have attracted great interest owing to their wide range of biological activity. This article describes the antifungal activity of synthetic δ-lactones against yeasts of the genus Candida spp. and dermatophytes (through the broth microdilution method), discusses the pathways by which the compounds exert this action (toward the fungal cell wall and/or membrane), and evaluates the toxicity to human leukocytes and chorioallantoic membrane (by the hen’s egg test-chorioallantoic membrane). Two of the compounds in the series presented broader spectrum of antifungal activity, including against resistant fungal species. The mechanism of action was related to damage in the fungal cell wall and membrane, with specific target action dependent on the type of substituent present in the δ-lactone structure. The damage in the fungal cell was corroborated by electron microscopy images, which evidenced lysed and completely altered cells after in vitro treatment with δ-lactones. Toxicity was dose dependent for the viability of human leukocytes, but none of the compounds was mutagenic, genotoxic, or membrane irritant when evaluated at higher concentrations than MIC. In this way, δ-lactones constitute a class with excellent perspectives regarding their potential applications as antifungals.

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.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Batista BG, Dalla Lana DF, Silveira GP, Sá MM, Ferreira M, Russo TVC, Canto RFS, Barbosa FAR, Braga AL, Kaminski TFA, de Oliveira LFS, Machado MM, Lopes W, Vainstein MH, Teixeira ML, de Andrade SF, Fuentefria AM (2017) Allylic selenocyanates as new agents to combat Fusarium species involved with human infections. ChemistrySelect 2:11926–11932

    Article  CAS  Google Scholar 

  • Berkow EL, Lockhart SR (2017) Fluconazole resistance in Candida species: a current perspective. Infect Drug Resist 10:237–245

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Burow ME, Weldon CB, Tang Y, Navar GL, Krajewski S, Reed JC, Hammons TG, Clejan S, Beckman BS (1998) Differences in susceptibility to tumor necrosis factor alpha-induced apoptosis among MCF-7 breast cancer cell variants. Cancer Res 44:4940–4946

    Google Scholar 

  • Carrasco H, Raimondi M, Svetaz L, Liberto MD, 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 17:1002–1024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • CLSI (2008a) Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved Standard – Third Edition. CLSI document M27-A3. Clinical and Laboratory Standards Institute, Wayne

    Google Scholar 

  • CLSI (2008b) Reference method for broth dilution antifungal susceptibility testing of filamentous fungi. Approved Standard – Second Edition. CLSI document M38-A2. Clinical and Laboratory Standards Institute, Wayne

    Google Scholar 

  • CLSI (2012) Reference method for broth dilution antifungal susceptibility testing of yeasts. Fourth Informational Supplement. CLSI document M27-S4. Clinical and Laboratory Standards Institute, Wayne

    Google Scholar 

  • Dahdah MJ, Scher RK (2008) Dermatophytes. Curr Fungal Infect Rep 2:8–86

    Article  Google Scholar 

  • Dalla Lana DF, Batista BG, Alves SH, Fuentefria AM (2016) Dermatophytoses: etiological agents, clinical forms, therapy and new perspectives of treatment. Clin Biomed Res 36:230–241

    Article  Google Scholar 

  • Eggimann P, Garbino J, Pittet D (2003) Epidemiology of Candida species infections in critically ill non-immunosuppressed patients. Lancet Infect Dis 3:685–702

    Article  PubMed  Google Scholar 

  • El Hage S, Lajoie B, Feuillolay C, Roques C, Baziard G (2011) Synthesis, antibacterial and antifungal activities of bifonazole derivatives. Arch Pharm (Weinheim) 344:402–410

    Article  CAS  Google Scholar 

  • Espinel-Ingroff A, Fothergill A, Peter J, Rinaldi MG, Walsh TJ (2002) Testing conditions for determination of minimum fungicidal concentrations of new and established antifungal agents for Aspergillus spp.: NCCLS collaborative study. J Clin Microbiol 40:3204–3208

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fernández-García R, de Pablo E, Ballesteros MP, Serrano DR (2017) Unmet clinical needs in the treatment of systemic fungal infections: the role of amphotericin B and drug targeting. Int J Pharm 525:139–148

    Article  CAS  PubMed  Google Scholar 

  • Ferreira M, Bisol TB, Conceição HP, Russo TVC, Bortoluzzi AJ, Sá MM (2017) One-pot synthesis of α-ylidene δ-lactones from functionalized allylic bromides in a water–isopropanol medium. Synthesis 49:667–676

    CAS  Google Scholar 

  • Frost DJ, Brandt KD, Cugier D, Goldman R (1995) A whole-cell Candida albicans assay for the detection of inhibitors towards fungal cell wall synthesis and assembly. J Antibiot 48:306–310

    Article  CAS  PubMed  Google Scholar 

  • Fuentefria AM, Pippi B, Dalla Lana DF, Donato KK, de Andrade SF (2018) Antifungals discovery: an insight into new strategies to combat antifungal resistance. Lett Appl Microbiol 66:2–13

    Article  CAS  PubMed  Google Scholar 

  • Ghannoum M (2016) Azole resistance in dermatophytes: prevalence and mechanism of action. J Am Podiatr Med Assoc 106:79–86

    Article  PubMed  Google Scholar 

  • Guinea J (2014) Global trends in the distribution of Candida species causing candidemia. Clin Microbiol Infect 20:5–10

    Article  PubMed  Google Scholar 

  • ICCVMA (Interagency Coordinating Committee on the Validation of Alternative Methods) (2010) The hen's egg test–chorioallantoic membrane (HET-CAM) test method. Research Triangle Park: National Toxicology Program. https://ntp.niehs.nih.gov/iccvam/docs/protocols/ivocularhetcam.pdf

  • Jackson JE (2003) A user’s guide to principal components. Wiley, Hoboken https://pdfs.semanticscholar.org/70a9/769e8027b48bce5a9ce3a82d0ce88e0f0730.pdf

    Google Scholar 

  • Janecka A, Wirebska A, Gach K, Fichna J, Janecki T (2012) Natural and synthetic α-methylenelactones and α-methylenelactams with anticancer potential. Drug Discov Today 17:561–572

    Article  CAS  PubMed  Google Scholar 

  • Kishimoto N, Sugihara S, Mochida K, Fujita T (2005) In vitro antifungal and antiviral activities α -and δ-lactone analogs utilized as food flavoring. Biocontrol Sci 10:31–36

    Article  CAS  Google Scholar 

  • Kuriyama T, Williams DW, Bagg J, Coulter WA, Ready D, Lewis MA (2005) In vitro susceptibility of oral Candida to seven antifungal agents. Oral Microbiol Immunol 20:349–353

    Article  CAS  PubMed  Google Scholar 

  • Martinez-Rossi N, Peres N, Rossi A (2008) Antifungal resistance mechanisms in dermatophytes. Mycopathologia 166:369–383

    Article  PubMed  Google Scholar 

  • Modranka J, Albrecht A, Jakubowski R, Krawczyk H, Zalski MR, Krajewska U, Janecka A, Wyre A, Zalska BR, Janecki T (2012) Synthesis and biological evaluation of α-methylidene-δ-lactones with 3,4-dihydrocoumarin skeleton. Bioorg Med Chem 20:5017–5026

    Article  CAS  PubMed  Google Scholar 

  • Montagner GFS, Sagrillo M, Machado MM, Almeida RC, Mostardeiro CP, Duarte MP, da Cruz IB (2014) Toxicological effects of ultraviolet radiation on lymphocyte cells with different manganese superoxide dismutase Ala16Val polymorphism genotypes. Toxicol in Vitro 24:1410–1416

    Article  CAS  Google Scholar 

  • Naglik JR, Richardsson JP, Moyes DL (2014) Candida albicans pathogenicity and epithelial immunity. PLoS Pathog 10:1–4

    Article  CAS  Google Scholar 

  • Ohkuma H, Naruse N, Nishiyama Y, Tsuno T, Hoshino Y, Sawada Y, Konishi M, Oki T (1992) Sultriecin, a new antifungal and antitumor antibiotic from Streptomyces roseiscleroticus. Production, isolation, structure and biological activity. J Antibiot 45:1239–1249

    Article  CAS  PubMed  Google Scholar 

  • Parker JE, Warrilow AG, Price CL, Mullins JG, Kelly DE, Kelly SL (2014) Resistance to antifungals that target CYP51. J Chem Biol 7:143–161

    Article  PubMed  PubMed Central  Google Scholar 

  • Pfaller MA, Rhomberg PR, Messer SA, Jones RN, Castanheira M (2015) Isavuconazole, micafungin, and 8 comparator antifungal agents’ susceptibility profiles for common and uncommon opportunistic fungi collected in 2013: temporal analysis of antifungal drug resistance using CLSI species-specific clinical breakpoints and proposed epidemiological cutoff values. Diagn Microbiol Infect Dis 82:303–313

    Article  CAS  PubMed  Google Scholar 

  • Pippi B, Lopes W, Reginatto P, Silva FEK, Joaquim AR, Alves RJ, Silveira GP, Vainstein MH, Andrade SF, Fuentefria AM (2018) New insights into the mechanism of antifungal action of 8-hydroxyquinolines. Saudi Pharm J 2018. https://doi.org/10.1016/j.jsps.2018.07.017.j

  • Rocha DFS, Sa LFR, Pinto ACC, Junqueira ML, Silva EM, Borges RM, Ferreira-Pereira A (2018) Characterization of an ABC transporter of a resistant Candida glabrata clinical isolate. Mem Inst Oswaldo Cruz 113:1–7

    Article  CAS  Google Scholar 

  • Romero-Guido C, Belo I, Ta TMN, Cao-Hoang L, Alchihab M, Gomes N, Thonart P, Teixeira JÁ, Destain J, Waché Y (2011) Biochemistry of lactone formation in yeast and fungi and its utilisation for the production of flavor and fragrance compounds. Appl Microbiol Biotechnol 89:535–547

    Article  CAS  PubMed  Google Scholar 

  • Santos RB, Júnior VL, Castro EVR, Balthar VO, Greco SJ (2009) Uso de métodos quimiométricos e mecânicoquanticos na análise de terpenóides e fenilpropanóides bioativos contra o Aedes aegypti. Orbital: Electron J Chem 4:273–288

    Google Scholar 

  • Silveira GP, Ferreira M, Fernandes L, Moraski GC, Cho S, Hwang C, Franzblau SG, Sá MM (2012) Allylic thiocyanates as a new class of antitubercular agents. Bioorg Med Chem Lett 22:6486–6489

    Article  CAS  PubMed  Google Scholar 

  • Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  PubMed  Google Scholar 

  • Tanaka H, Kageyama K, Yoshimura N, Asada R, Kusumoto K, Miwa N (2007) Anti-tumor and anti-invasive effects of diverse delta-alkyllactones: dependence on molecular side-chain length, action period and intracellular uptake. Life Sci 80:1851–1855

    Article  CAS  PubMed  Google Scholar 

  • Tavares LC (2004) QSAR: Hansch's approach. Quim Nova 27:631–639

    Article  CAS  Google Scholar 

  • Thomas P, Harvey S, Gruner T, Fenech M (2008) The buccal cytome and micronucleus frequency is substantially altered in Down’s syndrome and normal ageing compared to young healthy controls. Mutat Res 638:37–47

    Article  CAS  PubMed  Google Scholar 

  • Topliss JG (1996) Lipophilicity in drug action and toxicology (book review). J Med Chem 39:5287–5288

    Article  CAS  Google Scholar 

  • Umezawa I, Komiyama K, Oka H, Okada K, Tomisaka S, Miyano T, Takano S (1984) A new antitumor antibiotic, kazusamycin. J Antibiot 37:706–711

    Article  CAS  PubMed  Google Scholar 

  • Vieira JN, Feijó AM, Bueno ME, Gonçalves CL, Lund RG, Mendes JF, Villarreal JPV, Villela MM, Nascente OS (2018) Evaluation of the frequency of Candida spp. in hospitalized and non-hospitalized subjects. Braz J Biol 78:644–652

    Article  CAS  PubMed  Google Scholar 

  • Wani MC, Ronman PE, Lindley JT, Wall ME (1980) Plant antitumor agents. Synthesis and biological activity of camptothecin analogues. J Med Chem 23:554–560

    Article  CAS  PubMed  Google Scholar 

  • Wenderski TA, Stratton CF, Bauer RA, Kopp F, Tan DS (2015) Principal component analysis as a tool for library design: a case study investigating natural products, brand-name drugs, natural product-like libraries, and drug-like libraries. Methods Mol Biol 1263:225–242

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yamada T, Maeda M, Alshahni MM, Tanaka R, Yaguchi T, Bontems O, Salamin K, Fratti M, Monod M (2017) Terbinafine resistance of Trichophyton clinical isolates caused by specific point mutations in the squalene epoxidase gene. Antimicrob Agents Chemother 61:e00115–e00117

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yang EJ, Kim YS, Chang HC (2011) Purification and characterization of antifungal δ-dodecalactone from Lactobacillus plantarum AF1 isolated from kimchi. J Food Prot 74:651–657

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors are grateful to the Center of Microscopy and Microanalysis-UFRGS for the SEM.

Funding

This work was funded by the Brazilian agencies Coordination for the Improvement of Higher Education Personnel (CAPES, Finance Code 001), National Council for Scientific and Technological Development (CNPq), and Foundation for Research Support of the State of Rio Grande do Sul (FAPERGS; Edital PRONUPEQ 2016). G. P. Silveira received financial support from CAPES-UdelaR bilateral collaboration (grant no. 049-2013). A. M. Fuentefria, M. Apel, and M. M. Sá received PQ fellowships from CNPq.

Author information

Authors and Affiliations

  1. Laboratory of Applied Mycology, Department of Analysis, Faculty of Pharmacy, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Daiane F. Dalla Lana, Ânderson R. Carvalho & Alexandre M. Fuentefria

  2. Department of Molecular Biology and Biotechnology, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    William Lopes & Marilene H. Vainstein

  3. Biostatistics Unit, Research Group and Post-graduation, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS, Brazil

    Luciano S. P. Guimarães

  4. Laboratory of Biochemistry and Toxicology, Instituto Federal de Santa Catarina, Concordia, SC, Brazil

    Mário L. Teixeira

  5. Center for Studies in Biochemistry, Immunology, and Toxicology, Universidade Federal do Pampa, Uruguaiana, RS, Brazil

    Luis F. S. de Oliveira & Michel M. Machado

  6. Department of Raw Materials Production, Faculty of Pharmacy, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Saulo F. de Andrade

  7. Department of Chemistry, Universidade Federal de Santa Catarina, Florianopolis, SC, Brazil

    Marcus M. Sá & Theo V. C. Russo

  8. Department of Organic Chemistry, Institute of Chemistry, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Gustavo P. Silveira

Authors
  1. Daiane F. Dalla Lana
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ânderson R. Carvalho
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. William Lopes
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marilene H. Vainstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Luciano S. P. Guimarães
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mário L. Teixeira
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Luis F. S. de Oliveira
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Michel M. Machado
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Saulo F. de Andrade
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Marcus M. Sá
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Theo V. C. Russo
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Gustavo P. Silveira
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Alexandre M. Fuentefria
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Daiane F. Dalla Lana, Ânderson R. Carvalho, and William Lopes conceived, planned, and carried out the experiments. Marcus M. Sá, Theo V. C. Russo, and Gustavo P. Silveira synthesized and characterized the samples. Marilene H. Vainstein, Luciano S. P. Guimarães, Mário L. Teixeira, Luis F.S. de Oliveira, and Michel M. Machado assisted in the toxicological and statistical analyses and in obtaining the images from scanning electron microscopy. Alexandre M. Fuentefria and Saulo F. de Andrade contributed to the interpretation of the results and supervision of all the work. Daiane F. Dalla Lana took the lead in writing the manuscript and performed the general research. All authors provided critical feedback and helped shape the research, analysis, and manuscript.

Corresponding author

Correspondence to Daiane F. Dalla Lana.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. For this type of study formal consent is not required. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Additional information

Publisher’s note

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

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Dalla Lana, D.F., Carvalho, Â.R., Lopes, W. et al. Structure-based design of δ-lactones for new antifungal drug development: susceptibility, mechanism of action, and toxicity. Folia Microbiol 64, 509–519 (2019). https://doi.org/10.1007/s12223-018-00675-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12223-018-00675-y

Search

Navigation