Synergistic Effects of Caffeoylquinic Acid Compounds with Levofloxacin Against Uropathogenic : Insights from Molecular Dynamics Simulations, Antibiofilm, and Antimicrobial Assessments.

Urinary tract infections (UTIs), primarily caused by uropathogenic (UPEC), have high morbidity and recurrence rates. Resistance to levofloxacin hydrochloride (LEV), a commonly used treatment for UTIs, is increasingly problematic, exacerbated by biofilm formation mediated by interactions between cyclic di-GMP (c-di-GMP or CDG) and YcgR. In this study, we identified three caffeoylquinic acid compounds from -chlorogenic acid (CGA), sibiricose A5 (Si-A5), and 3--caffeoylquinic acid methyl ester (CAM)-that target YcgR through molecular docking. Biological assays revealed that combining these compounds with levofloxacin hydrochloride significantly enhanced antibacterial activity against standard UPEC strains in a concentration-dependent manner and clinically isolated UPEC strains. Notably, chlorogenic acid and sibiricose A5, when used with levofloxacin hydrochloride, enhanced intracellular c-di-GMP levels and swimming motility, significantly reduced YcgR gene expression, and effectively inhibited biofilm formation of UPEC at multiple time points. Additionally, molecular dynamics simulations elucidated the strong binding of these compounds to YcgR, underscoring the critical roles of residues, such as Arg118 and Asp145. This research serves as a foundation for tackling antibiotic resistance and developing innovative therapeutics for UTIs.