目前对大多数药物的作用机理和靶标知之甚少。随着药理学的进步，人们已经认识到小分子药物在体内往往具有多个作用靶标。明确药物的作用靶标，可以阐明药物的作用机理，通过构效关系优化药物的结构并提高药效；还能够预测可能出现的副作用，降低药物研发的风险。本项目拟开展高效率富集药物靶标蛋白的亲合色谱整体柱技术。对研究的小分子药物进行结构修饰后，作为功能化单体与交联剂在石英毛细管内同聚，就可以形成亲合整体柱。利用有机整体柱比表面大、传质阻力小，分离纯化步骤简单，对蛋白分离效率高的特点，提高对细胞中的药物靶标蛋白的富集效率。采用蛋白组学的质谱分析技术，鉴定被亲和整体柱富集的药物靶标；再用生化分析和分子生物学的方法对靶标蛋白进行验证。我们还将研究小分子药物的化学修饰和固定化方式对亲合基质富集靶标蛋白效率的影响，从而提高靶标蛋白亲合选择性并降低非特异性吸附。此项目将为新药发现的研究提供非常有用的技术。

Thus far, the knowledge on the action mechanism and targets of currently used small molecular drugs remains limited. With the development of pharmacology, it was realized that the small molecular drugs may have several action targets in the body. Identification of these drug targets is very helpful to reveal the action mechanism of the drugs, and subsequently to improve their efficacy by the following structure-activity relationship study. Moreover, it is also very valuable to predict the possible side-effect of the lead compounds prior to the clinic trial hence reduction of the failure risk of the drug develop project. The aim of the present proposal is to develop a new technique which can be used to effectively enrich the drug targets from the cell lysate. The proposed new technique is based on affinity chromatography with the monolithic capillary column. The small molecular drugs will be chemically modified so as to introduce a linker with a double bond terminal. The modified drug derivative can be copolymerized with other monomers and cross linker to form the monolith bed in the fused silica capillary. It can be predicted that the high separation performance of the monolith column will improve the efficiency of the drug target enrichment from the cell lysate. In combination with the mass spectrometry technology, the enriched proteins can be identified. Finally, such identified proteins will be validated with the biochemical analysis, such as western blotting. The effect of the chemical modification of the model drugs and the conditions used for monolith preparation will be systematically investigated so as to find a way to improve the affinity specific interaction of the affinity matrix to the drug target proteins, and to reduce the nonspecific adsorption. Our research will produce very useful tool for drug discovery.