腺苷脱氨酶抑制剂快速筛选模型的建立  

Rapid Drug Model for Screening Adenosine Deaminase Inhibitor

作　　者：董佩杰[1] 张新国[1] 刘英娟[1] 唐鹏[1] 苏媛 苏岷山 韩锡萍 邵男 

机构地区：[1]兰州理工大学生命科学与工程学院,甘肃省中藏药筛选与深加工重点实验室,兰州730050 [2]青海省西宁市药检所,西宁810000

出　　处：《中国现代应用药学》2015年第11期1301-1305,共5页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金(31360379);国家级大学生创新创业训练计划支持(091073117)

摘　　要：目的建立腺苷脱氨酶(adenosine deaminase,ADA)抑制剂的快速筛选模型。方法采用ADA酶,以腺苷为底物,2'-脱氧助间型霉素作为抑制剂验证反应体系,通过紫外分光光度法检测,优化反应条件,并对248种中药提取物及其内生菌发酵产物进行筛选。结果建立了最佳模型方案为:p H 7.5的PBS缓冲液体系,底物浓度为腺苷200μg·m L?1,ADA酶浓度55 IU·m L?1,连续检测腺苷在265 nm下OD值动态变化情况。此模型可以用于ADA酶抑制剂的快速筛选,且基于此模型共筛选获得23个有一定活性的粗提物。结论本实验建立的体系可以为ADA酶抑制剂的筛选提供简单易行、稳定、可靠的研究平台。OBJECTIVE To establish a rapid screening model for adenosine deaminase（ADA） inhibitor. METHODS Using the bovine spleen ADA as enzyme, adenosine as substrate, pentostatin as an inhibitor of the reaction system verification, an ultraviolet spectrophotometry method was used to screen for adenosine deaminase inhibitor after optimize its reaction conditions. RESULTS The best model scheme for p H was 7.5, phosphate buffer solution（PBS） was buffer system, the concentration of the substrate concentration of adenosine was at 200 μg·m L^-1, and the concentration of ADA enzymes was at 55 IU·m L^-1. The model could be used for rapid screening of ADA enzyme inhibitors and 23 activities were obtained from 248 kinds of Chinese herb extracts and fermentation products endophytes screening by the present model. CONCLUSION The present method is simple, stable and reliable for screening ADA inhibitor.

关 键 词：腺苷脱氨酶 抑制剂 筛选模型 

分 类 号：R965.1[医药卫生—药理学]