High-throughput screening of human soluble protein tyrosine phosphatase 1B inhibitors.
- Author:
Xiao-bin PANG
1
;
Xin-mei XIE
;
Shou-bao WANG
;
Guan-hua DU
Author Information
1. National Center for Pharmaceutical Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
- Publication Type:Journal Article
- MeSH:
Drug Evaluation, Preclinical;
methods;
Enzyme Inhibitors;
analysis;
pharmacology;
Escherichia coli;
metabolism;
High-Throughput Screening Assays;
methods;
Humans;
Inhibitory Concentration 50;
Protein Tyrosine Phosphatase, Non-Receptor Type 1;
antagonists & inhibitors;
metabolism;
Sensitivity and Specificity;
Vanadates;
pharmacology
- From:
Acta Pharmaceutica Sinica
2011;46(9):1058-1064
- CountryChina
- Language:Chinese
-
Abstract:
To screen potential human soluble protein tyrosine phosphatase 1B (PTP1B) inhibitors, a high-throughput screening (HTS) model in 384-well microplate with total volume of 50 microL was established. Recombinant PTP1B was cloned and expressed in E. coli. with its specific substrate 4-nitrophenyl phosphate disodium salt hexahydrate (PNPP). The HTS model was based on enzyme reaction rate with enhanced sensitivity and specificity (Z' = 0.78). A total of 24,240 samples were screened, among them 80 samples with inhibition greater than 70% were selected for further rescreening. Finally, six compounds with high inhibitory activity were identified, whose IC50 values were 21.58, 18.39, 15.37, 11.92, 37.27, and 36.61 microg x mL(-1), separately. The results indicated that the method was stable, sensitive, reproducible and also suitable for high-throughput screening.
