谷胱甘肽转移酶抑制剂的高通量筛选  

High throughput screening for glutathione S-transferase inhibitors

作　　者：张丹参[1] 张天泰[1] 李韶菁[1] 杜冠华[1] 

机构地区：[1]中国医学科学院中国协和医科大学药物研究所

出　　处：《药学学报》2008年第1期108-112,共5页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金重点资助项目(30630073).

摘　　要：To identify the inhibitor of glutathione S-transferase（GST）,a high-throughput screening method was established in a 384-well microplate with total 35 μL volume,and the absorbance at 340 nm is detected.The concentrations of substrates,CDNB and GST were determined by chromatometry.The optimal enzyme kinetics reaction time and temperature are 2 h and 30 ℃,respectively.The established model was evaluated by NaOCl,a known GST inhibitor,and the parameter Z′ was 0.77,which showed a high feasibility and stability of the assay.A total of 31 098 compounds were screened,of which 4 compounds were shown to inhibit GST activity,high inhibiting activity for their IC50 of GST inhibition was 3.94,4.05,74.85,and 77.41 mg·L-1,separately.The results indicated that the colorimetric method by using CDNB and GSH as substrate is stable,sensitive,reproducible and also suitable for high throughput screening.To identify the inhibitor of glutathione S-transferase （GST） , a high-throughput screening method was established in a 384-well microplate with total 35μL volume, and the absorbance at 340 nm is detected. The concentrations of substrates, CDNB and GST were determined by chromatometry. The optimal enzyme kinetics reaction time and temperature are 2 h and 30℃ , respectively. The established model was evaluated by NaOCl, a known GST inhibitor, and the parameter Z＇ was 0. 77 ,which showed a high feasibility and stability of the assay. A total of 31 098 compounds were screened, of which 4 compounds were shown to inhibit GST activity, high inhibiting activity for their IC50 of GST inhibition was 3.94, 4.05, 74.85, and 77.41 mg·L^-1, separately. The results indicated that the colorimetric method by using CDNB and GSH as substrate is stable, sensitive, reproducible and also suitable for high throughput screening.

关 键 词：谷胱苷肽转移酶 谷胱苷肽转移酶抑制剂 高通量筛选 

分 类 号：R965.1[医药卫生—药理学]