Establishment and optimization of an enzyme-cell based high-throughput screening platform for IDO1 inhibitors
10.16438/j.0513-4870.2019-0697
- VernacularTitle:基于酶学-细胞水平的IDO1抑制剂筛选模型建立与优化
- Author:
Fang-fang LAI
1
;
Ni-na XUE
1
;
Ming JI
1
;
Ting-ting DU
1
;
Ling LI
1
;
Li SHENG
1
;
Xiao-guang CHEN
1
Author Information
1. State Key Laboratory of Bioactive Substances and Functions of Natural Medicines/Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
- Publication Type:Research Article
- Keywords:
high-throughput;
tertiary screening platform;
indoleamine 2,3-dioxygenase 1;
inhibitor;
tumor immunity
- From:
Acta Pharmaceutica Sinica
2019;54(10):1868-1874
- CountryChina
- Language:Chinese
-
Abstract:
In this study, we used the tumor immunotherapy protein indoleamine 2,3-dioxygenase 1 (IDO1) as the target, and proposed an enzyme-cell-based tertiary IDO1 inhibitor high throughput screening platform. Firstly, the recombinant human IDO1 protein was expressed by genetic engineering and efficient IDO enzymatic screening system was established. Secondly, A172 cells stimulated with interferon-γ (IFNγ) or constructed plasmid which could highly express human IDO1 protein in HEK293 cells with transient transfection were used to construct the specific IDO1 cell based screening system. Finally, the effect of the compound on kynurenine and tryptophan in mouse plasma was determined by LC/MS/MS method on C57 mice, which could further verify the inhibitory effect of the selected compounds on IDO1 in vivo. The established and optimized enzyme-cell based screening model in this study can efficiently and effectively obtain IDO1 inhibitors in vitro, which lays a good foundation for the rapid development of clinical drugs. Procedures for animal study were performed with approval of the Animal Care and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College.
