靶向beclin-1小分子激动剂诱导三阴性乳腺癌细胞自噬性死亡和凋亡  

Discovery of a small-molecule activator of beclin-1 that induces autophagy-associated cell death and apoptosis in triple negative breast cancer

作　　者：赵荣演 靳文珂 姜星 袁兆鑫 刘博[2] 符雷蕾 ZHAO Rong-yan;JIN Wen-ke;JIANG Xing;YUAN Zhao-xin;LIU Bo;FU Lei-lei(School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China;State Key Laboratory of Biotherapy and Cancer Center,Sichuan University,Chengdu 610041,China)

机构地区：[1]西南交通大学生命科学与工程学院,四川成都610031 [2]四川大学生物治疗国家重点实验室,四川成都610041

出　　处：《药学学报》2021年第5期1369-1383,共15页Acta Pharmaceutica Sinica

基　　金：四川省应用基础研究项目(2020YJ0285)。

摘　　要：细胞自噬(autophagy)是一种溶酶体降解途径,在细胞内环境稳态、细胞发育、细胞免疫、抑制肿瘤、能量代谢、预防神经退行性变和延长寿命方面起着至关重要的作用。自噬的药理学激活可能是治疗癌症等某些人类疾病的有效方法。本研究基于beclin-1已有的晶体结构,联合变构位点识别方法、高通量虚拟筛选及体外活性评价,发现化合物10能激活自噬且具有良好的抗MDA-MB-231细胞增殖活性[半数抑制浓度(IC_(50)=8.25±1.53μmol·L~(-1))]。随后的分子对接、分子动力学模拟及免疫印迹研究结果表明,化合物10能够靶向激活beclin-1。体外研究表明,化合物10能够诱导MDA-MB-231细胞发生自噬相关性死亡。同时还发现化合物10激活自噬后,还伴随细胞凋亡的发生。本研究确定了候选化合物10作为开发有效的、具有选择性靶向beclin-1激活自噬的先导物,为进一步开发和优化用于临床治疗的靶向beclin-1激活自噬小分子药物提供依据。Autophagy is a lysosomal degradation pathway,and plays a crucial role in cellular homeostasis,development,immunity,tumor suppression,metabolism,prevention of neurodegeneration,and lifespan extension.Thus,pharmacological stimulation of autophagy may be an effective approach for preventing or treating certain human diseases and/or aging.Here,combined with allosteric site identification methods,high-throughput virtual screening,and in vitro activity evaluation,we found that compound 10 can activate autophagy and has good antiMDA-MB-231 cell proliferation activity(the half maximal inhibitory concentration IC50=8.25±1.53μmol·L-1).Subsequently,molecular docking,molecular dynamics simulation,and immunoblotting assay demonstrate that compound 10 can target and activate beclin-1.In vitro studies have shown that compound 10 can induce autophagyassociated cell death in MDA-MB-231 cells.In addition,it was found that compound 10 can induce apoptosis in MDA-MB-231 cells.Taken together,we identified the candidate compound 10 as an effective and selective targeting beclin-1 to activate autophagy as a lead compound,which provide a reference for further development and optimization of small molecule drugs targeting beclin-1 to activate autophagy for clinical treatment.

关 键 词：三阴性乳腺癌 BECLIN-1 小分子激动剂 自噬 凋亡 

分 类 号：R966[医药卫生—药理学]