Table1_Design, Synthesis, and Quorum Quenching Potential of Novel Catechol–Zingerone Conjugate to Find an Elixir to Tackle Pseudomonas aeruginosa Through the Trojan Horse Strategy.DOCX

To address the issue of multidrug resistance in Pseudomonas aeruginosa, a novel catechol–zingerone conjugate (1) linked via a non-hydrolyzable 1,2,3-triazole linker was synthesized and subjected to biological evaluation based on the Trojan horse strategy. To enhance the efficacy, catechol, a xenosiderophore, utilized by P. aeruginosa for iron assimilation, and the dietary phytochemical zingerone, known for its anti-virulent activity against Pseudomonas aeruginosa, were exploited in the present study. Theoretical validation of conjugate (1) was conducted by in silico molecular docking analysis to determine the interaction with outer membrane transport receptor PirA and quorum sensing signal receptors. In addition, nine-fold binding affinity of Conjugate (1) toward PirA (5FP2) in comparison to its natural ligand catechol with D-score −1.13 Å authenticated the designed Trojan horse drug. Conjugate (1) showed stronger anti-virulent activity than zingerone; hence, it exhibited a promising anti-biofilm efficacy as assessed by crystal violet assay and visualized by FESEM toward P. aeruginosa. Encouraging results against P. aeruginosa in terms of quorum sensing regulated virulence factors, motility phenotypes, and biofilm formation with no cell cytotoxicity and could help open hitherto unexplored possibilities of establishing Trojan horse drugs as a successful approach against multidrug resistance in P. aeruginosa.

为解决铜绿假单胞菌（Pseudomonas aeruginosa）的多重耐药性问题，本研究合成了一种通过非水解型1,2,3-三唑连接臂连接的新型儿茶酚-姜酮缀合物（1），并基于特洛伊木马策略（Trojan horse strategy）开展生物学评价。为提升药效，本研究选用了铜绿假单胞菌用于铁摄取的异源铁载体（xenosiderophore）——儿茶酚，以及已知对铜绿假单胞菌具有抗毒活性的膳食植物化学物姜酮。本研究通过计算机分子对接分析（in silico molecular docking analysis）对缀合物（1）进行理论验证，以明确其与外膜转运受体PirA及群体感应信号受体的相互作用。相较于天然配体儿茶酚（D评分为−1.13 Å），缀合物（1）对PirA（5FP2）的结合亲和力高出9倍，这证实了所设计的特洛伊木马药物的合理性。缀合物（1）的抗毒活性强于姜酮；经结晶紫染色法（crystal violet assay）评估、场发射扫描电子显微镜（FESEM）可视化显示，其对铜绿假单胞菌展现出优异的抗生物膜效能。该缀合物在群体感应调控的毒力因子、运动表型及生物膜形成方面对铜绿假单胞菌表现出令人鼓舞的抑制效果，且无细胞毒性，有望为将特洛伊木马药物开发为对抗铜绿假单胞菌多重耐药的有效策略开辟迄今尚未探索的新路径。