谷胱甘肽转移酶抑制剂的高通量筛选.pdfVIP

. 108. 药学学t/X Acta Pharmaceutica Sînîca 2008 ,43 (1) : 108 - 112 ·研9巳捕报· 谷脱甘肤转移酶抑制剂的茵通量筛选 张丹参l ， 2 ，张天泰l ，李韶菁l ，杜冠华I * (1.中国民学科学院、中网协和医科大学药物研究所，北京 100050; 2. 问北北方学院药学系，河北张家口 075000 ) 关键询:谷脱背肤转移酶;谷脱背肤转移酶抑制剂;高通最筛选 中圈分类号: R965. 1 文献标识码:A 文章编号: 0513 - 4870( 2008) 圳市0108 山 05 High throughput screening for glutathione S-transferase inhibitors ZHANG Dan-sh en 1.2, ZHANG Tian-tai , LI Shao-jing , DU Guan-hua (1. lnstitute 01 Materia Medica , Chinese Acαdemy 01 Medical Sciences and Peking Union Medical College , Beijing 100050 , China; 2. Department 01 Phαrmacy ， Hebei North University , Zhangjiαkou 075000 , China) Abstract: To identify the inhibitor of glutathione S-transferase (GST) , a high-throughput screening method was establìshed in a 384幡well microplate wìth total 35μL volume , and the absorbance at 340 nm ìs detected. The concentratìons of substrates , CDNB and GST were determìned by chromatometry. The optìmal enzyme kìnetics reaction time and temperature are 2 h and 30 C, respectively. The establìshed model was evaluated by NaOCI , a known GST ìnhìbitor , and the parameter Z was 0.77 ,which showed a hìgh feasibility and stability of the assay. A total of 31 098 compounds were screened , of whìch 4 compounds were shown to inhibit GST activity , high inhibiting activity for their IC so of GST inhibition was 3.94 , 4.05 , 74.85 , and 77.41 mg • L -1 , separately. The results indìcated that the colorìmetrìc method by usìng CDNB and GSH as substrate is stable , sensìtive , reproducible and also suitable for high throughput screening. Key words: glutathione S-transferase; glutathione S-transferase inhibitor; high throughput screening 谷脱甘肤转移酶 (gl