樟脑基缩氨基硫脲衍生物通过活性氧(ROS)介导的线粒体途径诱导人乳腺癌细胞的G2期阻滞和凋亡

Camphor-Based Thiosemicarbazone Analogues Induced G2 Cell Cycle Arrest and Apoptosis via Reactive Oxygen Species(ROS)-Mediated Mitochondrial Pathway in Human Breast Cancer Cells

以樟脑为原料合成了22个樟脑基缩氨基硫脲衍生物,通过1H NMR、13C NMR和HRMS对其结构进行了表征,并通过单晶X射线衍射测定了2-(3-(4-吡啶基)-1,7,7-三甲基二环[2.2.1]庚-2-亚基)肼硫代甲酰胺(3n)的晶体结构.通过3-(4,5-二甲基吡啶-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺苯基)-2H-四唑(MTS)法探索了这些衍生物对人乳腺癌细胞(MDA-MB-231)、人肺腺癌细胞(A549)和人多发性骨髓瘤细胞(RPMI-8226)三株肿瘤细胞的抗增殖活性以及对正常人细胞(GES-1)的细胞毒性.结果表明,这些衍生物都表现出较好的抗肿瘤活性,且对正常细胞GES-1毒性低(IC50>50μmol·L^-1).其中,2-(3-(4-蒽亚苄基)-1,7,7-三甲基二环[2.2.1]庚-2-亚基)肼硫代甲酰胺(3s)对MDA-MB-231细胞表现出最强的抗肿瘤活性[IC50=(3.90±0.04)μmol·L^-1].此外,初步的抗肿瘤机制结果表明,化合物3s可以通过细胞内活性氧(ROS)的增加和线粒体膜电位的破坏,诱导MDA-MB-231细胞的G2期阻滞和剂量依赖式凋亡,且通过凋亡相关蛋白的变化对实验结果进行了验证.

22 novel camphor-based thiosemicarbazone derivatives were synthesized using camphor-based thiosemicarbazone as material and their structures were determined by 1H NMR,13C NMR and HRMS.The crystal structure of 2-(3-(pyridin-4-ylmethylene)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-ylidene)hydrazinecarbothioamide(3n)was determined by single crystal X-ray diffraction.The derivatives were screened in vitro for anticancer activities against human breast cancer cell line(MDA-MB-231),human lung adenocarcinoma cell line(A549),human multiple myeloma cell line(RPMI-8226)and toxicity against a normal human cell line(GES-1)by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS).It was found that majority of the tested analogs showed moderate to significant antitumor activity against selected cancer cell lines.Noticeably,2-(3-(anthracen-9-ylmethylene)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-ylidene)-hydrazinecarbothioamide(3s)exhibited selective anti-tumor activities against MDA-MB-231 cells(IC50=3.90±0.04μmol·L^-1)and low toxicity to GES-1 cells(IC50>50μmol·L^-1).In the process of exploring the underlying mechanism of 3s,it was found that compound 3s could cause G2 phase arrest and apoptosis in MDA-MB-231 cells by overproduction of intracellular reactive oxygen species and collapse of mitochondrial membrane potential.The measured results were confirmed by western blot assay.