二肽基肽酶-4(DPP-4)在胰岛素分泌中起重要负调节作用，是治疗Ⅱ型糖尿病的重要靶点，寻找安全有效的DPP-4抑制剂是抗糖尿病药物研究的关键之一。中药复杂体系中活性成分的快速筛选与分离是发现和创制新药的一条重要途径。本项目拟以全二维LC为基础建立快速高通量在线识别DPP-4抑制剂的联用新技术2D LC-EAD-MS，以有效提高色谱分辨率和筛选准确性；系统研究中药提取物中DPP-4抑制剂；并针对CCC分离效率低、峰容量有限的缺点建立高效快速的2D CCC用于目标活性单体化合物的导向性分离。建立一套基于2D LC-EAD-MS/2D CCC技术的中药复杂体系中DPP-4抑制剂的快速筛选和制备新方法。通过系统的体外活性筛选，应用现代光谱方法结合化学方法鉴定结构，同时进行构效关系分析，以期发现新型DPP-4抑制剂，为找寻和开发抗糖尿病新药奠定基础，同时促进天然产物分析分离新技术的探索与发展。

As an important target of treating type II diabetes, dipeptidyl peptidase-4(DPP-4) plays a significant inhibitory role in insulin production. Therefore, seeking safe and effective DPP-4 inhibitors becomes one of the keys to developing new antidiabetes durgs. The research on active constituents of natural products (NP) is known as one of important pathways of developing new drugs. In this proposal, based on comprehensive two-dimensional (2D) liquid chromatography (LC), an on-line screening technique 2D LC-EAD-MS will be built up for high-throughput and fast identification of DPP-4 inhibitors from complex samples to effectively improve the resolution of LC and accuracy of screening. DPP-4 inhibitors from extracts of some Traditional Chinese Medicines (TCM) would be systematically investigated. Meanwhile, to address drawbacks of relatively low separation efficiency and limited peak capacity in conventional Counter-Current Chromatography (CCC), a more effective on-line 2D CCC system will be built up for target-guided separation of the compounds with DPP-4 inhibitory activity. Based on the hyphenated 2D LC-EAD-MS/2D CCC technique, a new methodology for rapidly screening and separating DPP-4 inhibitors from complex TCM will be established. By systematically screening the activity of those DPP-4 inhibitors in vitro, elucidating their structures by modern spectral and chemical methods, and analyzing their structure-activity relationship, new DPP-4 inhibitors were expected to be found, which will pave way for seeking and developing new antidiabetic drugs and facilitate the exploration of new technologies for analyzing and separating NP.