亲和超滤质谱技术在中药活性成分筛选中的研究进展  

Recent Advances of Affinity Ultrafiltration Mass Spectrometry in Screening Active Components of Traditional Chinese Medicine

作　　者：周慧[1] 王义民 郑重[2] 刘舒[2] 刘志强[2] ZHOU Hui;WANG Yi-min;ZHENG Zhong;LIU Shu;LIU Zhi-qiang(Department of Chemistry and Pharmacy,Zhuhai College of Jilin University,Zhuhai 519041,China;Changchun Center of Mass Spectrometry,Changchun Institute of Applied Chemistry,Chinese Academy of Sciences,Changchun 130022,China)

机构地区：[1]吉林大学珠海学院,广东珠海519041 [2]中国科学院长春应用化学研究所,吉林长春130022

出　　处：《质谱学报》2018年第6期641-652,共12页Journal of Chinese Mass Spectrometry Society

基　　金：国家自然科学基金项目(81403077);吉林省科技发展计划国际科技合作项目(20170414008GH);广东省自然科学基金博士启动项目(2014A030310494)资助

摘　　要：亲和超滤质谱技术是20世纪90年代中期发展起来的一种快速、简单、有效的药物小分子发现模式。该技术利用配体与受体之间特异性结合,通过超滤装置快速筛选活性小分子化合物,再结合液相色谱-质谱联用技术(LC-MS),鉴定活性成分结构。亲和超滤质谱技术集药物活性成分筛选、结构鉴定于一体,尤其适用于从复杂体系中筛选潜在的活性小分子化合物。近年来,针对中药发挥药理作用具有多组分、多靶点的重要特点,亲和超滤质谱技术已被广泛用于从中药提取物中筛选与特定蛋白靶点相结合的小分子活性物质,对阐明中药药效的物质基础和以活性成分作为先导化合物的新药开发具有重要意义,是对传统药物发现方法的有利补充。本工作综述了该技术在中药活性成分筛选中的原理、特点、应用进展,以及对未来的展望。Affinity ultrafiltration mass spectrometry(AUF-MS)technology is a rapid,simple and effective method for the discovery and development of small active molecule in traditional Chinese medicine(TCM).This technology was initially developed in the middle of 1990s,which was introduced to target-oriented drug discovery.Through the ligand-receptor specific binding characteristics,the affinity ultrafiltration device can facilitate the rapid screening of small-molecule ligands from the complex extracts,and high performance liquid chromatography-mass spectrometry(HPLC-MS)assists in the structural identification of potentially active small drug molecules.Thus,AUF-MS has become a powerful tool for identifying bioactive molecules from complex chemical matrixes.The general workflow of AUF-MS is not complicated:depending on the molecular weight cut-off of the semi-permeable membrane,the ligand-bound protein complexes are retained by the membrane under centrifugal force to separate the bound and unbound components of the analyte.Bound ligands are eluted from the ultrafiltration membrane by destabilizing the target-ligand complex with an organic solvent or pH change,and then the ligands are consequently subjected to analysis by LC-MS to identify and characterize the low molecular weight compounds that interact with target molecules.The classic strategies of screening active ingredients from natural product,such as the high-throughput screening methods based on ultraviolet fluorimetric or radioactive detection,the traditional phytochemical screening procedure(i.e.isolation,structure elucidation and bioactivity test),are not able to meet the urgent need of contemporary pharmacology research due to its time consumption and a high rate of false positive or negative results.In comparison with those methods,AUF-MS is easier to be operated without unnecessary isolation or purification of inactive compounds and directly screen bioactive ingredients from the Chinese herb.Also,only modest amounts of target proteins are required,which are no

关 键 词：亲和超滤质谱(AUF-MS) 中药 靶蛋白 活性成分筛选 高通量 

分 类 号：O657.63[理学—分析化学]