Screening of new human carbonic anhydrase Ⅱ(hCA Ⅱ) inhibitors
- VernacularTitle:人碳酸酐酶Ⅱ(hCA Ⅱ)抑制剂筛选技术及应用
- Author:
Zhonghai XIAO
;
Ruifeng DUAN
;
Wenyu CUI
;
Hai WANG
- Publication Type:Journal Article
- Keywords:
high-altitude illness;
acetazolamide;
inhibitor;
carbonic anhydrase;
isozyme;
prokaryotic expression;
screening technique
- From:
Chinese Pharmacological Bulletin
2010;26(2):182-185
- CountryChina
- Language:Chinese
-
Abstract:
Aim To provide practical method for screening human carbonic anhydrase Ⅱ(hCA Ⅱ) inhibitors in drug discovery.Methods hCA Ⅱ protein was obtained from induced BL21(DE3) E.coli containing plasmid pET-28b-hCA Ⅱ.The hCA Ⅱ activity was detected under pH 7.6 and 25℃ by its esterase activity which could decompose PNPA to increase the photoabsorption at 348 nm. After the assay conditions were finally selected, 35 new compounds were tested.Results A practical method for screening hCA Ⅱ inhibitors was successfully constituted by using recombinant hCA Ⅱ protein expressed in E.coli as the source of hCA Ⅱ enzyme.10 new compounds had better inhibitory effect and 9 new compounds had the same inhibitory effect on hCA Ⅱ compared with acetazolamide.Conclusions The hCA II inhibitor screening technique constituted in this work possesses advantages of being reliable, rapid, and practical. 19 new compounds are worth further research for developing high efficiency and low side effect drugs used for high-altitude illness.
