Integrative computational-experimental discovery of α-hederin as a multi-mechanistic, low-toxicity antifungal agent targeting Candida albicans CYP51

Introduction Infections caused by pathogenic fungi such as Candida albicans have led to a continuous increase in disease morbidity and mortality, underscoring the urgent need for safer and more effective antifungal therapies. Current antifungal drugs are limited by toxicity and resistance, highlighting the necessity for innovative discovery strategies. Objectives The aim of this study was to identify novel antifungal compounds using a integrative computational approach targeting C. albicans CYP51 (14-α-sterol demethylase). Specifically, we sought to screen the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database for potential antifungal candidates, evaluate their therapeutic potential, and establish a computational-experimental framework to expedite antifungal drug development. Methods We employed an integrative computational approach-targeting Candida albican s CYP51 (14-α-sterol demethylase) via molecular docking and pharmacophore modeling-to screen the TCMSP database. The two natural small molecules obtained were subjected to in vivo and in vitro antifungal experiments, and their antifungal mechanisms were analyzed through molecular docking and molecular dynamics simulation. Result This yielded two novel antifungal candidates: α-hederin and elemenin, with minimum inhibitory concentrations (MICs) of 32 μg/mL and 16 μL/mL against C. albicans , respectively. α-Hederin demonstrated superior therapeutic potential, showing low cytotoxicity in 293 T, Raw264.7, and KB cells. It effectively inhibited hyphal formation, biofilm formation, and cell surface hydrophobicity in vitro . In a murine oral candidiasis model, α-hederin outperformed fluconazole by reducing fungal burden, inhibiting hyphal invasion, and preventing tongue adhesion. Molecular dynamics simulations revealed that α-hederin forms a more stable complex with CYP51 than fluconazole, engaging additional hydrogen bonds, alkyl interactions, and carbon-hydrogen bonds. Conclusion This study advances antifungal drug discovery by validating CYP51 as a high-value target for structure-guided screening, identifying α-hederin as a low-toxicity, multi-mechanistic antifungal agent and establishing a computational-experimental framework for rapid antifungal development.