氮杂环基金属配合物、复合物的构筑及其抗肿瘤活性研究

The complexes have special spatial architecture and target, showing different anti-tumor activities by contrast to the conventional chemotherapy. This project is designed to construct metal complexes by the reaction of nitrogen heterocyclic ligands and transition metal salts, such as Zn, Cu, etc. In addition, by applying the transition metal salts and ligands described above, this project is supposed to synthesize functionalized C60-based complexes or functionalized graphene oxide-based hybrid compounds. The relationship between the structure of complexes and performance will be studied by changing the ligand environment around the metal center; also, the corresponding functions of metal centers in each complex will be confirmed by the replacement of metal center. Complexes with inhibitory activity for digestive, reproductive system cancer cell lines will be investigated, and the experimental methods, such as optics, single staining, Annexin V/PI double staining, ROS, Western blot, etc., will be utilized to clarify the apoptosis signal transduction pathway of the tumor cell caused by complexes; the effect of their cytotoxicity should be explicited after importing functionalized C60 or graphene oxide into the nitrogen heterocyclic metal complexes; the target protein will be determined by 2D electrophoresis, mass spectrometry, Western blot, RT-PCR; the action mode between complexes and the target at molecular level will be further revealed by circular dichroism, NMR, X-Ray, dynamics simulation and other approaches; tumor cell apoptosis mechanism induced by complexes will be elucidated.

配合物具备独特的立体结构和靶向性，从而表现出与传统化疗药物不同的抗肿瘤活性。本项目旨在以氮杂环配体和Zn、Cu等过渡金属盐构筑配合物，以及使用氮杂环配体与功能化的C60协同组装配合物或制备功能化氧化石墨烯-配合物的复合物。通过改变金属中心外围的配体环境，研究配合物结构与性能的关系；置换金属中心，明确金属离子在配合物表达相应功能方面的作用。筛选对消化、生殖系统肿瘤细胞的增殖有抑制活性的配合物，运用光学、PI单染、Annexin V/PI双染、ROS、Western blot等实验手段，阐明其诱导肿瘤细胞凋亡的信号转导通路；明确功能化的C60、氧化石墨烯分别导入氮杂环金属配合物后对细胞毒性的影响；利用2D电泳、质谱，Western blot、RT-PCR确定靶点蛋白，进一步通过圆二色谱、核磁、X-Ray、动力学模拟等方法，揭示配合物与靶点在分子水平的作用方式，及其诱导肿瘤细胞凋亡的作用机制。

研究配合物抗肿瘤的作用机制，对设计、开发出具有系统毒性低、靶点作用强、抗肿瘤效果及选择性好的目标化合物具有重要意义。本项目合成了多个苯并咪唑衍生物的氮杂环配体，采用常规化学合成、溶剂热等方法构筑了系列具有不同结构特点的配合物，并分析了有机配体对配合物结构的调控。筛选出了多个具有良好溶解性能及抗肿瘤活性的配合物。利用紫外滴定、荧光、CD光谱考察体外DNA与配合物的作用，并研究了配合物对DNA模板转录的抑制；通过ICP-MS分析了细胞对具有不同结构特点配合物的摄入，及配合物在细胞内的分布，研究了配合物结构与细胞摄入、细胞毒性的关系，证明了配合物在细胞内与遗传物质DNA的作用是其细胞毒性的重要因素之一；利用彗星实验评估了配合物对核DNA的损伤；运用流式技术分析了配合物对细胞周期，线粒体膜电位，胞内活性氧水平、Ca2+离子浓度，细胞凋亡、坏死的影响。通过上述研究，证明了具有共轭芳香基团的氮杂环配体构筑的配合物与DNA具有较强的相互作用，多核金属配合物由于金属中心-配体间的协同作用，能有效的抑制肿瘤细胞的增殖；分析了具有不同结构特点的配合物诱导肿瘤细胞凋亡的作用机制。本项目发表标注基金资助的SCI期刊论文9篇。