A comprehensive in vitro and in silico assessment of eugenol glycoconjugates against azole and amphotericin B resistant Rhizopus spp

Mol Biol Rep. 2025 Jun 12;52(1):589. doi: 10.1007/s11033-025-10673-2.

Abstract

Background: Rhizopus spp. is a major cause of mucormycosis, a severe infectious disease with high morbidity and mortality. Treatment is challenging due to rising antifungal resistance. Glycosylation is a crucial technique for enhancing the properties of phenolic compounds like eugenol. The present study aims to examine the antifungal efficacy of eugenol glycoconjugates against azole and amphotericin B-resistant Rhizopus isolates.

Methods and results: Out of 50 soil samples, 12 isolates belonging to Mucorales were obtained, of which 7 were identified as Rhizopus spp. via 18S ITS sequencing. Antifungal susceptibility testing (AST) revealed that all Rhizopus isolates were resistant to amphotericin B (MIC > 1 µg/mL). Most isolates also showed resistance towards posaconazole (MIC > 1 µg/mL) and itraconazole (MIC > 2 µg/mL). AST of eugenol glycoconjugate (coded 6g) showed efficacy against resistant Rhizopus isolates, with MIC values ranging from 6.25 µg/mL to 25 µg/mL. Flow cytometry confirmed its fungicidal activity, correlating with MIC data. Compound 6g significantly reduced conidial germination within 24 h and exhibited no cytotoxicity on A549 lung cancer cells. In-silico analysis revealed a negative binding affinity of compound 6g for the spore coat protein CotH3, which could be a potential antifungal target.

Conclusion: Compound 6g could be an potential antifungal molecule against resistant Rhizopus spp, which requires further studies.

Keywords: Antifungals; Eugenol; Glycoconjugates; Mucormycosis; Resistance; Virulence.

MeSH terms

  • A549 Cells
  • Amphotericin B / pharmacology
  • Antifungal Agents / pharmacology
  • Azoles / pharmacology
  • Computer Simulation
  • Drug Resistance, Fungal / drug effects
  • Eugenol* / chemistry
  • Eugenol* / pharmacology
  • Glycoconjugates* / chemistry
  • Glycoconjugates* / pharmacology
  • Humans
  • Microbial Sensitivity Tests
  • Mucorales / drug effects
  • Mucormycosis / drug therapy
  • Mucormycosis / microbiology
  • Rhizopus* / drug effects
  • Rhizopus* / genetics
  • Triazoles / pharmacology

Substances

  • Eugenol
  • Antifungal Agents
  • Amphotericin B
  • Glycoconjugates
  • Azoles
  • posaconazole
  • Triazoles