吲哚胺2,3-双加氧酶(IDO)的色胺酮类抑制剂筛选及其体外抗肿瘤作用  

Screening of tryptanthrin derivatives of indoleamine 2,3-dioxygenase (IDO) inhibitor and their antitumor activity in vitro

作　　者：杨双双[1] 杜丽莎[1] 李豪男 杨青[1] 

机构地区：[1]复旦大学生命科学学院生化系,上海200433 [2]金日成综合大学生命科学系平壤

出　　处：《复旦学报（医学版）》2014年第2期149-155,共7页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(30873153)~~

摘　　要：目的评价色胺酮衍生物吲哚胺2,3-双加氧酶(indoleamine 2,3-dioxygenase,IDO)的抑制活性,并研究其作为IDO抑制剂的抗肿瘤作用。方法采用基因工程手段表达、纯化重组人IDO(rhIDO),建立IDO活性检测体系;以色胺酮衍生物作为对象,进行IDO抑制活性初筛,对抑制类型、半数抑制浓度以及抑制常数进行测定;构建高表达人IDO的pcDNA3.1(+)-hIDO转染HEK 293细胞,评价色胺酮衍生物在细胞水平上的IDO抑制活性;采用MTT比色法考察色胺酮衍生物3对人非小细胞肺癌A549细胞的生长抑制作用。结果 6个被测色胺酮衍生物均具有IDO抑制活性,且细胞水平上的抑制效力高于酶活水平,抑制效力均优于目前通用的IDO抑制剂1-甲基色氨酸(1-methyl-tryptophan,1-MT)。色胺酮衍生物3作为最强的IDO抑制剂,其Ki值为0.161μmol/L。MTT实验结果显示,色胺酮衍生物3显著抑制A549细胞生长,IC50为8.77μmol/L。结论色胺酮衍生物是一类新型高效的IDO抑制剂,在体外对A549细胞具有较强的抗肿瘤活性。Objective To evaluate the inhibitory activity of tryptanthrin derivatives of indoleamine 2,3-dioxygenase （IDO）,and to explore the effects on human nonsmall cell lung cancer A549 cell growth.Methods IDO activity assay system was established with recombinant human IDO （rhIDO）,which was expressed and purified by the technology of genetic engineering.Based on the system,we detected the IDO inhibitory activity of tryptanthrin derivatives,and determined the inhibition type,kinetic parameters and IC50 values.We also evaluated the IDO inhibitory effects of tryptanthrin derivatives on HEK 293 cells transfected with pcDNA3.1（＋）hIDO.Inhibitory activity of tryptanthrin derivatives on A549 cell growth inhibition was analysed by MTT assay.Results Six tryptanthrin derivatives displayed higher IDO inhibitory activity than 1-methyl tryptophan （1-MT）,which is the commonly used IDO inhibitor.IC50 values of tryptanthrin derivatives,obtained from the HEK 293 cellbased assay,were much lower than that from the enzyme assay.Compound 3 was the best inhibitor and had Ki value of 0.161 μmol/L.Treating A549 cells with 3 remarkably inhibited the cell growth,and its IC50 value was 8.77 μmol/L.Conclusions Tryptanthrin derivatives are novel and potent IDO inhibitors,and can significantly inhibit human non-small cell lung cancer A549 cell growth in vitro.

关 键 词：活性检测 色胺酮衍生物 吲哚胺2 3-双加氧酶(IDO) 抗肿瘤作用 

分 类 号：Q331[生物学—遗传学]