硫辛酸-香豆素偶联物合成及抗肿瘤活性研究  

Synthesis and anticancer activity of lipoic acid coumarin conjugate

作　　者：戴一[1,2] 冯艳 陈天芹 DAI Yi;FENG Yan;CHEN Tian-qin(College of Pharmaceutical Sciences,Anhui Xinhua University,Hefei 230088,China;Department of Chemistry,University of Science and Technology of China,Hefei 230026,China)

机构地区：[1]安徽新华学院药学院,合肥230088 [2]中国科学技术大学化学系,合肥230036

出　　处：《天然产物研究与开发》2020年第3期379-384,372,共7页Natural Product Research and Development

基　　金：安徽省教育厅自然科学研究重点项目(KJ2019A0873);安徽新华学院制药工程教学团队建设项目(2017jxtdx08)。

摘　　要：硫辛酸为一内源性二硫化物,通过线粒体途径抑制多种肿瘤细胞的增殖,而对正常细胞影响甚微;香豆素为一具有荧光的天然产物,广泛应用于生物成像。本实验设计合成了一个新颖的硫辛酸-香豆素偶联物,该偶联物具有多功能。香豆素作为荧光报告器,硫辛酸作为活性药物。当该偶联物进入肿瘤细胞,由于肿瘤细胞中的水解酶把偶联物中的酯键水解,释放出硫辛酸和荧光分子香豆素而发出荧光。采用噻唑兰(MTT)法评价其对肿瘤细胞的增殖抑制活性,活性氧检测试剂盒(DCFH-DA)检测其对活性氧的清除作用,流式细胞术检测其诱导的细胞凋亡及荧光显微检测细胞内成像。实验结果发现该偶联物的细胞毒性相比于母体化合物硫辛酸提高了一个数量级,进一步研究发现该偶联物可清除肿瘤细胞增殖与生长所需的活性氧,同时该偶联物诱导的凋亡显著高于其他组药物。荧光显微镜监测该偶联物在给药6 h后发出的荧光最强,说明该偶联物具有示踪药物水解释放的功能。Lipoic acid is an endogenous disulfide compound capable of suppressing growth of various cancer cell lines via mitochondria-targeted mode of action,while non-transformed primary cells are hardly affected.Coumarin,a natural product with fluorescence,was one of widely used imaging reagents.We present here,the design,synthesis and in vitro biological assessment of a novel lipoic acid-coumarin conjugate.This conjugate is a multifunctional molecule composed of a coumarin moiety as a fluorescent reporter and lipoic acid as a model active drug.After entering tumor cells,the conjugate can release lipoic acid and concomitantly fluorescence intensity increases,due to hydrolysis of ester bonds between lipoic acid and coumarin by hydrolytic enzyme in cancer cells.MTT assay was used to investigate its inhibitory activity on tumor cells.Reactive oxygen species(ROS) removal was detected by DCFH-DA kit.Flow cytometry was used to detect apoptosis.Fluorescence microscopy was used to detect intracellular imaging.The results showed that compared with lipoic acid,the in vitro anticancer activity of the conjugate increased by an order magnitude.Further research revealed that the conjugate can scavenge reactive oxygen species,an important substances for proliferation and growth of cancer cells.Apoptosis induced by the conjugate was significantly stronger than those of other tested compounds.Fluorescence microscopic experiments reveal that the fluorescence intensity reaches its maximum at 6 h,indicating drug release can be readily monitored at the cellular level.

关 键 词：硫辛酸 香豆素 偶联物 抗肿瘤活性 

分 类 号：R914.5[医药卫生—药物化学]