Targeting fungal biofilms: Design, synthesis and evaluation of pyrrolidino-hydrazinyl-thiazole derivatives via in vitro, transcriptomic and in silico approaches.

The increase in the drug resistance in fungal infections, particularly those involving biofilm-forming species like Candida albicans, presents a global challenge to public health. The biofilm forming abilities of C. albicans contributes to high rates of morbidity and treatment failure, necessitating the development of novel therapeutic agents. This study reports the design and green synthesis of novel pyrrolidino-substituted hydrazinyl-thiazole derivatives and investigates their potential as fungal biofilm inhibitors, focusing on compounds 4h, 4k, and 4l. These compounds demonstrated potent, dose-dependent biofilm inhibition against C. albicans, with 4k and 4l showing highest biofilm disruption as revealed by fluorescent-based assay. Apart from biofilm, 4k and 4l also prevented hyphal morphogenesis, as confirmed by FESEM and live-cell imaging. Gene expression analysis revealed that treatment with these compounds led to significant downregulation of key virulence and biofilm-associated genes, including ALS3, HWP1, EFG1, and RAS1. In-silico molecular docking and dynamics simulations further elucidated the mechanism, showing that the compounds exhibit a high binding affinity and stable interaction with adenylate cyclase (AC), a key enzyme in the fungal morphogenesis pathway.