液相色谱-酶亲和检测联用技术研究进展

Research progress on liquid chromatography-enzyme affinity detection combined technology

液相色谱-酶亲和检测(high performance liquid chromatography-enzyme affinity detection,HPLC-EAD)联用技术是近年来逐渐建立的,根据酶对具有某一相同立体结构的底物具有亲和性,进而出现竞争性抑制现象的原理,用于筛选复杂体系中对某种酶具有亲和性的底物的一种新技术。本文调研了与该联用技术相关的大量文献,并从该技术的概述、应用和展望这3个方面进行综述。经过分析与总结发现这种技术虽然起步较晚,但是发展迅速,目前多用于药物代谢酶的抑制剂,中药活性成分的筛选和药物筛选。细胞色素P450酶、黄嘌呤氧化酶、α-葡萄糖苷酶等生物酶都在人体内发挥着十分重要的作用,通过筛选能够与这些酶高效结合的底物,进而研究其具体结合模型以及在体内的作用机制,是药物研发的重要手段。由于酶亲和技术本身存在的一些还未解决的缺陷,比如安全性与有效性、渗漏与变性、成本较高的问题,因而在一定程度上限制了它的发展与大规模应用。但是由于它的亲和特性,这一技术仍然在体内重要的代谢酶抑制剂的筛选、复方中药产生临床疗效的主要活性成分群的筛选等方面具有重要的理论意义和应用价值。

High performance liquid chromatography-enzyme affinity detection （ HPLC-EAD ） combined technology is a new technique gradually developed in recent years. It has been used to screen enzyme substrate since the enzyme substrate has a similar three-dimension structure with the enzyme, then the substrate can compete for the enzyme thus emerging competitive inhibition. In this paper, a great deal of literatures related to this technology was surveyed, and the overview, application and prospect of this technology were reviewed. After analysis and summarization we found that although this technology started late, it was developed rapidly and mostly used in the screening of drug metabolic enzyme inhibitors and active ingredients of traditional Chinese medicine screening and drug screening. Cytochrome P450 enzymes, xanthine oxidase, oL -glucosidase and other enzymes play a very important role in the human body, and screening the substrates that can combine the enzyme efficiently and study the specific binding model and mechunanisms in vivo is particularly important. Because of the unsolved defects of enzyme affinity technology, such as unsafety and in effectiveness, leakage and denaturation and high cost, the development of enzyme affinity technology is limited to a certain extent. But because of its affinity characteristics, this technique is still important in screening the metabolic enzyme inhibitors, the main active ingredient group producing clinical efficacy from compound traditional Chinese medicine, and it has important theoretical significance and application values.