High throughput screening for glutathione S-transferase inhibitors.
- Author:
Dan-Shen ZHANG
1
;
Tian-Tai ZHANG
;
Shao-Jing LI
;
Guan-Hua DU
Author Information
1. Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
- Publication Type:Journal Article
- MeSH:
Dinitrochlorobenzene;
chemistry;
Drug Evaluation, Preclinical;
methods;
Enzyme Inhibitors;
analysis;
Glutathione;
chemistry;
Glutathione Transferase;
antagonists & inhibitors;
Substrate Specificity
- From:
Acta Pharmaceutica Sinica
2008;43(1):108-112
- CountryChina
- Language:Chinese
-
Abstract:
To identify the inhibitor of glutathione S-transferase (GST), a high-throughput screening method was established in a 384-well microplate with total 35 microL 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 degrees C , 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 x 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.
