细菌生物被膜（BF）是导致耐药行为的重要原因。群体感应抑制剂（QSI）通过拮抗细菌自身的群感系统来抑制BF的形成进而避免耐药性的产生，已成为抗耐药菌治疗的全新手段。多样性的天然产物是获取QSI的重要来源，然而现有QSI筛选策略都是基于单一靶点进行，不符合QS系统网络特征，容易导致QSI失效甚至引起新的抗性。本研究拟基于网络药理学原理，选取铜绿假单胞菌（PA）中与BF形成密切相关的三种QS信号节点蛋白LasR/RhlR/MvfR，建立固定化多靶点亲和整体色谱阵列-纳流液相色谱-质谱在线筛选方法，寻找天然产物中新型多靶点QSI活性成分，并通过体外活性验证，结合虚拟筛选手段进行对比评估。本项目旨在提供一种“筛选、分离、鉴定”一体化的多靶点QSI活性成分研究方法，提高QSI药物筛选自动化程度及效率；为揭示QSI抗耐药菌的科学内涵，开发安全有效的抗菌新药提供理论依据。

The formation of biofilm (BF) is one of the most important causes of bacterial resistance behavior. Quorum sensing inhibitors (QSIs), which could interfere with quorum sensing system and therefore inhibit BF formation without causing resistance, is becoming one of the innovated treatment against drug-resistance bacteria. Natural product is an important source for discovering new QSIs. However, current strategies and methods for screening QSIs from natural products are limited to single-target, which overlooked the network features of QS system, and easily led to interference failure and even new resistance. According to the principle of network pharmacology, three target proteins i.e. LasR, RhlR, and MvfR, are considered as the most closely related to Pseudomonas aeruginosa (PA) BF formation. In this research, a monolithic column array will be developed for the first time by combining LasR/RhlR/MvfR-affinity onto polymer-based monolithic support. In order to screen multi-target QSIs from TCM extracts, an on-line platform, which integrates LasR/RhlR/MvfR-affinity monolithic column array, nano-HPLC, and mass spectrometry (MS), will be established. The anti-biofilm and anti-bacterial activity of the screened active ingredients will be verified by means of in vitro activity tests. Moreover, a comparative study between the proposed strategy and the computer virtual screening approach will be also carried out. This research will be useful for elucidating the scientific connotation of TCM against drug-resistant bacteria. It can not only provides a powerful tool for screening novel QSIs, but also shows great meaning in elucidating theoretical basis for the development of novel, safe, and effective drugs against resistant bacteria.