亲和色谱法筛选中药中血管紧张素转化酶抑制剂  

Screening of angiotensin converting enzyme inhibitors from traditional chinese Medicines by affinity chromatography

作　　者：黄燕[1] 田清清[2] 李谭瑶[2] 陈波[2] 姚守拙[1,2] 

机构地区：[1]湖南大学化学生物传感与计量学国家重点实验室,长沙410082 [2]湖南师范大学化学生物学及中药分析教育部重点实验室,长沙410081

出　　处：《中国科学（B辑）》2009年第8期760-766,共7页Science in China(Series B)

基　　金：国家重点基础研究发展计划(批准号:2006CB504701);国家自然科学基金(批准号:20875028)资助

摘　　要：以壳聚糖微球为载体、戊二醛(glutaraldehyde,GA)为交联剂对血管紧张素转化酶(Angiotensin converting enzyme,ACE)进行固定化.用固化的ACE作为亲和介质,利用血管紧张素转化酶抑制剂(Angiotensin converting enzyme inhibitor,ACEI)与ACE之间的亲和作用,结合高效液相色谱对亲和前后的体系进行检测,比较两者各组分色谱峰的差异,以此实现快速筛选复杂体系中的ACE抑制剂.应用赖诺普利(Lisinopril)、九肽抑制剂、依那普利(Enalapril)、培哚普利(Perindopril)、卡托普利(Captopril)等已上市的ACEI对方法进行验证,反映方法具有高度选择性.将方法应用于中药地龙及山楂筛选,发现共有5个组分与ACE有亲和作用,并且都能抑制ACE酶活性,它们对酶活性抑制的IC50值在0.45～4.62μg/mL范围.通过对亲和方法重现性考察,6次测定的相对标准偏差小于1%,说明方法可靠.提出的亲和色谱-色谱指纹差异法非常适合于从中药及天然产物等复杂混合物库中快速筛选靶点活性物质.An angiotensin converting enzyme （ACE） immobilized affinity chromatography was applied to screen angiotensin converting enzyme inhibitors （ACEIs） from traditional Chinese medicines （TCMs） based on the enzyme-inhibitor interaction. ACE immobilized on chitosan microspheres with glutaraldehyde （GA） as cross-linking reagent was used as stationary phase of the affinity chromatography. The components, which can be adsorbed on the immobilized ACE, were identified as the lead compounds of ACEIs. The high efficiency and selectivity of this method was confirmed by using some commercial available antihypertensive drugs, i.e., lisinopril, nonapeptides in- hibitor, enalapril, perindopril and captopril, as model ACEIs. Five components from two typical TCMs with obvious ACEI activities, Pheretima （Dilong） and Fructus Crataegi （Shanzha）, were screened out by affinity adsorption. All of these five components were proved with ACEI activities, and IC50 values of the components ranged from 0.45 to 4.62 ktg/mL. In addition, the relative standard deviation （RSD） of six repetitive determinations was lower than 1%. Thus, the proposed affinity interaction/chromatographic differences assay is suitable for rapidly screening of target active components from complicated matrix such as TCMs and natural products.

关 键 词：中药 血管紧张素转化酶抑制剂 筛选 亲和色谱 固定化酶 

分 类 号：TQ461[化学工程—制药化工] TS201.21[轻工技术与工程—食品科学]