仿生催化产生一氧化氮的级联酶组装体的构建及其抗血栓应用研究

Nitric oxide plays an extensive role in the expansion of blood vessels, regulation of blood pressure, and inhibition of platelet adhesion. Supplement of endo- and exo- genous nitric oxide in blood has become an essential approach to the prevention and treatment of thrombosis, hypertension and other cardiovascular diseases. In this project, a new biomimetic approach was proposed for the artificial catalytical generation of nitric oxide messenger based on the cascade enzyme assemblies. The oxidoreductase and peroxidase will be combined as cascade enzymes for the catalytical generation of nitric oxide in the presence of endogenous arginine, hydrogen peroxide as the intermediate product. The cascade enzymes will be assembled together as nanostructure assemblies simultaneously loading with imaging agent, targeting molecules and intelligent response macromolecules, and guarantee the nitric oxide release in a continuous, steady and controllable manner. The cell toxicity and biological effect of the biomimetic assemblies will be investigated, and the biomimetic assemblies will be further exploited as a nitric oxide donor for the prevention and treatment of thrombosis, and other cardiovascular diseases. This study holds great promise to provide new methods and technologies for the nitric oxide supplement, therapy of cardiovascular diseases. And this project is also an innovative subject that is expected to promote the development of chemical biology and cardiovascular medicine in the cross field of modern biomimetic chemistry and materials assembly.

一氧化氮具有扩张血管、调节血压、抑制血小板粘附等作用，补充供给内外源性一氧化氮、提高血液一氧化氮浓度是血栓、高血压等心血管疾病预防和治疗的一种重要措施。本项目模拟一氧化氮合酶功能，构建仿生催化产生一氧化氮的级联酶组装体，将为一氧化氮的生物供给提供一种新的模式。拟选择氧化还原酶和过氧化物酶构建级联反应酶对，以内源精氨酸等生物分子作为底物，双氧水作为酶反应中间体，仿生催化生成一氧化氮分子；拟通过载体组装与生物修饰，构建智能响应与靶向识别的级联酶组装体，以期实现一氧化氮持续、稳定与可控的生物供给；进一步将探讨仿生催化级联酶组装体的细胞毒性与生物效应，在分子水平、细胞与活体组织水平探讨组装体在抗血栓等心血管疾病方面的预防与治疗作用。本项目将为一氧化氮的生物供给及其心血管应用提供新的技术平台，这将是一项在仿生催化技术和材料组装技术等前沿交叉领域推动化学生物学与心血管医学发展的创新性课题。

一氧化氮具有扩张血管、调节血压、抑制血小板粘附等作用，补充供给内外源性一氧化氮、提高血液一氧化氮浓度是血栓、高血压等心血管疾病预防和治疗的一种重要措施。本项目模拟一氧化氮合酶功能，构建了仿生催化产生一氧化氮的级联酶组装体，比如人工凝集体囊泡合成细胞。以内源分子为底物，级联酶催化持续产生一氧化氮。进一步在分子、细胞与活体水平探讨了组装体的生物效应和一氧化氮生理效应，产生的一氧化氮具有心血管舒张效应。研究工作具体包括：1）催化生成NO的级联酶设计与筛选；2）仿生催化级联酶组装体的构建；3）仿生催化组装体的生物效应考察；4）仿生催化组装体的抗血栓应用研究。通过该项目，发表SCI学术论文34篇，包括Nat. Chem.、J. Am. Chem. Soc.、Angew. Chem.，大会报告6次，申请与授权发明专利合计4项，培养毕业 研究生10名。本项目为一氧化氮的生物供给及其心血管应用提供了新的技术平台，仿生催化技术和材料组装技术结合，在化学生物学与心血管医学领域具有潜在应用前景。