Theoretical and Experimental Investigations of 2,2-Dimethyl-1,3-dioxolane-4-methanamine-imprinted Polymers as AI-2-Inspired Surrog-ate-Recognition Materials for the Modulation of Quorum-Sensing-Regulated Phenotypes.

Quorum sensing (QS) is a bacterial communication mechanism that regulates collective behaviors such as virulence, motility, and biofilm formation. Consequently, materials capable of interacting with molecules associated with QS signaling pathways are of increasing interest as potential tools for studying and modulating bacterial communication processes. In this study, molecularly imprinted polymers (MIPs) were designed and synthesized using (R/S)-2,2-dimethyl-1,3-dioxolane-4-methanamine, an autoinducer-2 (AI-2)-inspired surrogate template to investigate molecular recognition of structures relevant to bacterial signaling systems. Five bulk MIPs were prepared by thermal radical polymerization using methacrylic acid, 4-vinylbenzoic acid, itaconic acid, trifluoromethacrylic acid, and acrylic acid as functional monomers. The resulting polymers were evaluated in terms of adsorption capacity, imprinting factor, and selectivity toward the template and structurally related compounds using liquid chromatography coupled with mass spectrometry (LC-MS)-based binding studies. Among the synthesized materials, the polymer prepared from methacrylic acid exhibited the highest specificity, with an imprinting factor of 3.10 at 50 μg L-1 and up to 19.14 at 2 μg L-1 of template concentrations, indicating strong concentration-dependent recognition. Computational modeling using density functional theory, molecular mechanics, and molecular dynamics simulations was employed to elucidate the molecular interactions governing template recognition and cavity formation. Theoretical analyses revealed that multiple hydrogen bonding, hydrophobic interactions, and electrostatic contacts contribute to stable analyte-polymer binding, supporting the experimentally observed selectivity. Selectivity experiments demonstrated preferential adsorption of the template and its closest structural analogue relative to several structurally related amine-containing compounds. Biological evaluation using the Chromobacterium violaceum model revealed that both molecularly imprinted and non-imprinted polymers reduced violacein production to varying degrees, indicating modulation of a QS-regulated phenotype under the experimental conditions employed. Collectively, these findings demonstrate the feasibility of using molecular imprinting strategies to generate polymeric materials capable of recognizing an AI-2-inspired surrogate template and influencing a QS-regulated biological response, highlighting their potential for development as innovative materials for antivirulence strategies against bacterial infections.