IDO1天然小分子抑制剂的筛选及抗肿瘤作用研究  

Screening of natural small molecule IDO-1 inhibitors with anti-tumor role

作　　者：庞伟强 高磊 窦玥莹 刘璐[1] 张娜[1] 刘晓嘉 刘阳 杨兆勇[1] 宋丹青[1] 邓洪斌[1] PANG Wei-qiang GAO Lei DOU Yue-ying LIU Lu ZHANG Na LIU Xiao-jia LIU Yang YANG Zhao-yong SONG Dan-qing DENG Hong-bin(Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College Beijing 100050, China Jilin Institute for Drug Control Changchun 130000, China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所,北京100050 [2]吉林省药品检验所,吉林长春130000

出　　处：《药学学报》2017年第9期1416-1423,共8页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上资助项目(81473248);中国医学科学院重大协同创新项目-重大前沿研究(2016-12M-1-011)

摘　　要：PCR扩增人吲哚胺2,3-双加氧酶1(indoleamine 2,3-dioxygenase 1,IDO1)基因启动子上游区域1 245 bp基因片段,将其插入到p GL4.20-basic载体中构建了p GL4-IDO1-luc荧光素酶重组质粒。基于双荧光素酶报告基因方法建立IDO1抑制剂筛选模型,筛选能够下调肿瘤细胞中IDO1表达的天然活性小分子化合物。采用MTT、Western blotting和乳酸脱氢酶(LDH)等方法探讨阳性化合物的抗肿瘤作用及其对IDO1的调控机制。化合物川楝素(toosendanin,NS-180)能显著下调IFN-γ诱导的肿瘤细胞中IDO1表达。在A549细胞中,NS-180可抑制STAT1和STAT3的磷酸化,从而下调IFN-γ诱导的IDO1蛋白表达。LDH释放实验表明,NS-180可促进NK细胞对A549细胞的杀伤作用。综上所述,筛选获得的天然小分子NS-180是一类新型高效的IDO1抑制剂,可能成为靶向IDO1的肿瘤免疫治疗候选药物。Fragments of the human indoleamine 2,3-dioxygenase 1（IDO1） gene 5＇-UTR（untranslated 1 245 bp region） promoters were amplified by PCR and cloned into p GL4.20 vector in the construction of reporter vector p GL4-IDO1-luc. A549 cells were transfected with the constructed plasmid and IDO1 inhibitor screening model was established with dual-luciferase reporter assay. Based on the model, we screened natural small molecules which could down-regulate the expression of IDO1 on tumor cells. The anti-tumor activities were examined by MTT, Western blotting and lactic dehydrogenase（LDH） release assays. Toosendanin（NS-180） down regulated the IDO1 expression and inhibited IFN-γ-induced STAT1 and STAT3 phosphorylation in A549 cells. Moreover, NS-180 significantly increased the cytotoxicity of co-cultured NK cells on A549 cells in LDH release assays. In summary, NS-180 is a novel and potent IDO1 inhibitor, which has an antitumor activity for cancer immunotherapies.

关 键 词：吲哚胺2 3-双加氧酶1 启动子 荧光素酶 川楝素 

分 类 号：R965.1[医药卫生—药理学]