@article{APS8215,
author = {Wei HU and Hui DONG and Yue-zhong LI and Xi-tao HU and Guan-jun HAN and Yin-bo QU},
title = {A high-throughput model for screening anti-tumor agents capable of promoting polymerization of tubulin in vitro},
journal = {Acta Pharmacologica Sinica},
volume = {25},
number = {6},
year = {2016},
keywords = {},
abstract = {AIM:
To establish a high-throughput model for screening anti-tumor agents capable of promoting the polymerization of tubulin in vitro.
METHODS:
Tubulin was prepared in different purity for two screening steps. The first step was a high-throughput screening (HTS) for a set of 1500 samples using the GTP-containing tubulin and the end-reading method. The second step was performed on 119 hits from the first screening by a kinetic assay with GTP-lacking tubulin.
RESULTS:
The HTS for 1500 samples was accomplished in less than 3 h. From the screening, 108 samples were identified with >20 % promotion activity at 10 mg/L. Five of 108 were further confirmed by the kinetic assay using the purified tubulin subsequently. Three of the hit compounds were Epothilone A or its analogs, the other two compounds had new structures with a common pharmacophore for cytotoxic natural products that stabilize microtubules. In an MTT test, the five selected samples from the screening showed a minimal IC(50) at 0.28+/-0.06 nmol/L to Hela cells.
CONCLUSION:
The two-step screening method is a high-throughtput, cost-effective, and efficient approach to identify microtubule-stabilizing agents.},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/8215}
}