基于新型功能化磁性纳米粒与LC-NMR/MS技术靶向筛选绵头雪莲花COX-2抑制剂的研究

Traditional Chinese Medicine (TCM) is a multi-component system with abundant diversities in chemical structures, which make the isolation and purification of bioactive components responsible for the pharmaceutical effects extremely difficult. Therefore, there has been longly and highly demand for the targeted isolation of desired constituents from TCM. In recent years, the development of functionalized nanotechnology has provided novel ideas and methods for this challengeable task. In this project, based on our previous study, we incorporated cyclooxygenase-2 (COX-2) to the surface of magnetic nano particles (MNPs) to construct functionalized MNPs targeted to COX-2 inhibitors. Using these MNPs as probes, combining with the LC-SPE-NMR/MS integrated technique as the detection tools, we have built a facile, fast and highly specific methodology for the targeted screening of bioactive small molecules especially with trace contents from TCM. The extract/metabolites of traditional anti-inflammatory herb "Snow Lotus" will be screened for COX-2 inhibitors using this method. The anti-inflammatory effects and safety of the obtained constituents will be assessed by both cell and animal tests, and their pharmacological mechanism will also be elucidated. The present project provides a novel and efficient screening platform with the advantages in selective separation, fast characterization, activity re-evaluation and mechanism elucidation. It possesses promising fortune not only in the discovery of active components/leading compounds from natural resources, but also in the study of relationship among ingredients-metabolism-potency/safety of TCM, and the basic research in the detection of trace metabolites of TCM, the signal transduction and the function of bioactive enzymes/protein.

中药具有组分多、分离难的特点，靶向性分离方法在中药活性成分的筛选研究中具有迫切需求。近年来，功能化纳米技术的发展为解决中药成分的靶向分离提供了新的思路。本项目在前期研究基础上，以磁性纳米粒（MNPs）为载体，以人环氧合酶-2（COX-2）为靶酶，通过偶联载体与靶酶，制备出靶向分离COX-2抑制剂小分子的新型功能化纳米粒（COX-2-MNPs），结合液相色谱-核磁共振波谱/质谱联用（LC-NMR/MS），构建一种方便、快捷、选择性强的COX-2抑制剂筛选方法，用于绵头雪莲花活性成分的靶向分离和识别，并在细胞/动物模型上阐明活性成分的功效机理和安全性。本项目将创建集中药活性成分靶向分离、结构表征与活性筛选为一体的技术平台，不仅可为加速新活性成分或先导化合物的发现提供重要的方法支撑，而且可在揭示中药成分-代谢-功效/安全的相关性、中药微量活性物质检测、酶/蛋白功能等基础研究领域发挥重要作用。

靶向性分离方法在中药活性成分的筛选研究中具有迫切需求，功能化纳米技术的发展为解决中药成分的靶向分离提供了新的思路。本项目在前期研究基础上：1）以磁性纳米粒（MNPs）为载体，以人环氧合酶-2（COX-2）为靶酶，通过偶联载体与靶酶，制备了靶向分离COX-2抑制剂小分子的新型功能化纳米粒（COX-2-MNPs），并结合液相色谱-核磁共振波谱/质谱联用（LC-NMR/MS），构建了一种方便、快捷、选择性强的COX-2抑制剂筛选方法；2）利用所建方法，靶向分离和识别了绵头雪莲花活性成分，发现了绵头雪莲花的抗炎物质基础scopoletin、syringin及其1:1配伍，弥补了现有抗炎药的不足；3）阐明了中药绵头雪莲花选择性抑制COX-2的抗炎机理，展现了传统中医药理论、资源在当代疾病防治领域的科学内涵和价值。本项目创建了集中药活性成分靶向分离、结构表征与活性筛选为一体的技术平台，不仅为加速新活性成分或先导化合物的发现提供了重要的方法支撑，而且可在揭示中药成分-代谢-功效/安全的相关性、中药微量活性物质检测、酶/蛋白功能等基础研究领域发挥重要作用。