新型稀土金属配合物的设计、合成及其作为TRPV4通道阻断剂的生物活性研究

Rare earth metals show various biological activity. TRPV4 is a broadly expressed Ca2+ permeable cation channel in the vanilloid subfamily of transient receptor. Studies show that altering the intracellular calcium concentration by TRPV4 receptor profoundly affects physiological function of organism. Recent findings reported indicate that TRPV4 is stimulated during distension of the bladder urothelium and could contribute to abnormal overactivity of the bladder in pathological conditions. In addition, reduction of TRPV4 channel activity strongly correlated with improvements in the overall progression of other different diseases, but unfortunately, there are few clinically approved drugs that target TRPV4 channels. ..This study is to design and synthesis rare earth matal complexe of substituent amino acid ester cyano-benzoimidazol derivatives and to investigate their activity and specificity as a TRPV4 blocker. Electrophysiological Recordings and measurement of intracellular calcium concentration will be measured in parallel experiments with a monochromator-based imaging system consisting of a Polychrome IV monochromator and a charge-coupled device camera in the present of synthetic cyano-benzoimidazol blocker/reported blocker. Taken together described-above measurement of TRPV4 and other TRP subfamily channel, extensive efforts will be undertaken to understand the molecular structure of action, improve the biological activity and specificity of TRPV4 blocker, and promote their clinical use based on the novel TRPV4 blocker. Therefore, it is to be hoped that more TRPV4 channel information will be achieved due to the novel blocker, and is possible to represent a novel therapy for different diseases.

稀土金属元素具有多种生物学活性，人工合成的稀土配合物既可增强其生理活性又可降低其生理毒性。瞬时受体势通道之一TRPV4通道作为最近新发现的分布广泛的钙离子摄入途径，在生理功能完成方面发挥重要作用。研究表明多种病症与TRPV4通道的过度激活紧密相关，选择性阻断TRPV4通道对于新药研发和临床治疗具有重要意义。本课题以稀土为配位离子，以氰基苯并咪唑为起始物，通过多步反应合成一系列取代氨基酸酯修饰的氰基苯并咪唑类配合物，利用细胞转染技术考察目标产物介导下TRPV4的表达效率及TRPV4过表达受阻断的细胞形态，通过膜片钳技术研究目标产物介导的TRPV4通道膜电压状况，引入活细胞钙离子浓度荧光检测技术检测目标产物对胞内钙离子浓度的影响，考察对TRPV4通道的阻断效率和选择性。建立配位离子、配体结构与阻断效率和选择性的关系，拓展稀土金属的生物学用途，为开发新型选择性TRPV4通道阻断剂奠定基础。

本项目按计划通过多步反应合成一系列取代氨基酸酯修饰的氰基苯并咪唑类配合物，在合成了目标有机配体之后，尝试了这些配体与稀土金属离子的配合，发现配合能力相对较差，生物活性较差、毒性高。其中大部分合成的有机物与金属配合后溶解性差，难于用于生物活性测试。考虑到苯并咪唑类化合物具有较为优良的生物活性，课题研究中拓展了多类衍生物，如氨基嘧啶丁烯酮衍生物、六氢氮杂卓-4-基氧基苯甲酰胺类化合物、氨基咪唑偶联吡啶酮衍生物并对其生物性质进行了研究。因此，本课题根据目前研究结果和文献调研情况，合成了氨基嘧啶丁烯酮衍生物、六氢氮杂卓-4-基氧基苯甲酰胺类化合物、氨基咪唑偶联吡啶酮衍生物、氨基嘧啶丁烯酮衍生物、硫代磷酸酯衍生物等，并通过1H NMR， 13C NMR， MS等手段对产物进行表征和结构确定，对其生物性质进行了研究。为这类杂环类化合物的生物应用提供了理论指导意义。相关研究成果发表SCI论文2篇，申请发明专利10项。