系列双功能型DNA切割体系的构筑及抗癌活性研究

The efficiency and specificity of metalloenzyme originate from the cooperative roles between the metalmediated catalysis at the first coordination sphere and the non-covalent interactions at the secondary coordinationsphere．In view of the inefficiency and non-specificity of traditional chemical nucleases, our project intends to design some kinds of bifunctional chemical nucleases, which process two DNA binding sites: recognition center and cleavage center. The rational design of two centers will enhance the binding abilities of target compounds and reflect the synergistic effect of two centers. The project will adopt the molecular biology measures to investigate the interaction mechanism between bifunctional compound and nucleic acid molecule and elaborate the nature and regularity of the synergistic effect. By means of the molecular dynamics simulation method and quantum chemistry calculation method, we will theoretical analyze the innate character of the interaction between the target compound and nucleic acid (especially the sequence specific oligodeoxynucleotide). The above studies will provide the theoretical and experimental basis to explore chemical nuclease with high efficiency and DNA specificity. In addition, we will filter some compounds with potential anticancer activity at the cellular level, and analyze the cell apoptosis process. Our project is expected to obtain some efficient specific chemical nucleases and several compounds with potential anticancer activities, which provide the valuable information for the development of complex-type anticancer drugs.

金属酶催化的高效性和选择性来源于第一配位环境金属催化中心和第二配位环境非共价相互作用力的协同作用。基于此，针对传统化学核酸酶切割效率低下、特异性差的问题，本项目从配体的设计角度出发，将非共价相互作用因素引入结合中心，实现金属切割中心和结合中心的分离，形成具有潜在协同效应的双功能中心。本项目将采用分子生物学的方法，探讨双功能化合物与核酸分子的作用机制，阐明化合物功能中心之间的协同效应的规律和本质；通过分子力学模拟和量子化学计算方法，从理论上分析化合物与核酸（尤其是特定序列的寡聚脱氧核苷酸）结合的特点和规律，为研发高效、具有特异结合性能的化学核酸酶提供实验和理论依据。另外，本项目还将从细胞层次对化合物进行抗癌活性筛选，寻找诱导细胞凋亡的靶点，对相关作用靶点和作用机制进行分析，为开发低毒、高效的配合物型抗癌药物提供有价值的信息。

本课题从配体的设计角度出发， 选择系列新型多吡啶类、多喹啉类、三联吡啶类和手性席夫碱类配体；在此基础之上，合成数十个金属配合物，对其结构和光谱特性进行了详细的表征。本项目从分子水平上探讨化合物与核酸或蛋白等生物大分子之间的作用规律，系统研究了化合物对核酸的切割机理及构效关系。在化学核酸酶活性研究基础之上，本项目还从细胞层次对部分化合物进行抗癌活性筛选，寻找诱导细胞凋亡的靶点，对相关作用靶点和作用机制进行分析，为开发低毒、高效的配合物型抗癌药物提供有价值的信息。此外，项目组选择类Salen 型配体，成功构筑了系列3d，3d-4f系列金属配合物，并对其结构和磁学性质展开深入探讨。通过课题组成员的共同努力，项目组基本完成计划所定的目标和内容，达到预定目标和水平，取得了一定的成绩；本项目发表相关学术论文15篇，获得国家发明专利3项。