IDO1 及 TDO 抑制剂体外筛选模型的建立及应用

	IDO1 及 TDO 抑制剂体外筛选模型的建立及应用; Establishment and application of IDO1 and TDO inhibitors screening model in vitro
	杨坤
导师	赵勤实，刘将新
摘要	L-tryptophan is an essential amino acid for the human body and cannot be synthesized by ourself. It is involved in human immune regulation, nerve function regulation and intestinal homeostasis. Tryptophan is mainly metabolized through the kynurenine pathway, indoleamine-2,3-dioxygenase 1 (IDO1) and tryptophan-2,3-dioxygenase (TDO) are the key rate-limiting enzyme to its metabolic pathway. The abnormally high expression of IDO1 and TDO in tumor tissues causes the exhaustion of tryptophan and the increase of metabolites such as kynurenine in the tumor microenvironment, which can inhibit the proliferation and activation of T cells and play an important role in tumor immune escape. Excessive activation of these two enzymes can also lead to diseases such as neurological disorders and neurodegenerative diseases. At present, there has been a lot of research on the correlation between IDO1 and TDO and cancer, neurodegenerative diseases and other diseases, but there is still a lot of challenge for the development of related inhibitors. In this dissertation, we selected IDO1 and TDO as new drug targets, constructed recombinant plasmids, and obtained the prokaryotic expression system of IDO1 protein and TDO protein. Using various purification methods, recombinant proteins with higher purity were obtained. A screening model for IDO1 and TDO inhibitors was established, and the conditions of the reaction system were explored and optimized. As for a result, this screening model is stable, reliable and effective. It can achieve a high-through and high-efficiency screening and increase the speed of discovering potential active compounds. Then we evaluated more than 300 compounds by the enzyme assay. Three nature compounds were discovered to be potent IDO1 inhibitors, and two as potent TDO inhibitors. A series of tanshinoneIIA derivatives were later synthesized. The structure-activity relationship of tanshinoneIIA derivatives with IDO1 inhibitory activity was investigated. At the same time, four derivatives were synthesized starting from berberine, and the derivatives were evaluated for activity using the established model. One compound with dual selectivity for IDO1 / TDO was found. In summary, this article established an in vitro enzymatic screening model of IDO1 and TDO inhibitors, and screened out compounds with better activity, which also provided a basis for further structural modification and structure-activity relationship exploration.
	2020-05
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74204
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	杨坤. IDO1 及 TDO 抑制剂体外筛选模型的建立及应用, Establishment and application of IDO1 and TDO inhibitors screening model in vitro[D],2020.

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