酰肼类衍生物抗流感病毒活性及其机制研究

Influenza viruses cause seasonal epidemics and pandemic outbreaks associated with significant morbidity and mortality, and a huge cost. Currently, the application of anti-influenza drugs is restricted due to the poor efficacy, side effects and drug resistance. Because of the rapid variation of influenza virus and the difference of individual response, the vaccine can only give limited protection. New influenza virus inhibitors with an entirely different mode of action are urgently required. The influenza polymerase complex is widely recognized as a key drug target, given its critical role in virus replication and high degree of conservation among influenza A and B viruses. We have found that the compounds containing diarylhydrazides skeleton can inhibit influenza virus replication strongly by using reporter system of RNA polymerase of influenza virus. In order to enhance the antiviral activity and reduce toxicity we will prepare new derivatives by modifying the leading compound of hydrazide. In this project we will assess the antiviral activity of the new derivatives in vitro and in vivo. And we will execute further experiments to elucidate the roles and molecular mechanism of the hydrazide compounds in anti-influenza virus, which will lay the foundation for further screening and development of anti-influenza drugs and enrich our knowledge about antiviral research system.

流感病毒能引起季节性流行病或全球范围大流行，每年大约导致25万到50万人死亡，是人类健康面临的严重威胁。目前主要的抗流感药物药效有限、毒副作用强及存在耐药性，且相关疫苗的研发无法跟上病毒变异的步伐，因此寻找和研制新型抗流感病毒药物在流感防控中至关重要。流感病毒聚合酶因其特异性已成为药物研发的热门靶点。我们前期采用流感病毒聚合酶报告系统发现含二芳基酰肼骨架的化合物对流感病毒有较强的抑制活性。本项目拟以酰肼类化合物为先导化合物，通过对其苯环上的取代基团进行替换和改造制备系列新衍生物，改善理化特性、降低毒性并增强抗流感病毒活性。通过验证新衍生物体外及体内的抗流感病毒活性，并深入揭示其抑制流感病毒聚合酶的分子机制，为进一步研发抗流感新药奠定重要基础并提供理论依据。

甲型流感病毒具有很高的抗原性变异性，且已批准的抗流感药物极其有限，因此开发用于临床治疗和预防流感暴发的新型抗流感药物势在必行。前期我们通过甲型流感病毒聚合酶报告基因实验筛选获得一个含酰肼骨架化合物具有抗甲型流感病毒活性。通过该项目的实施我们合成50多个新衍生物并测试了这些化合物的抗甲型流感病毒活性和细胞毒性。基于这些化合物的构效关系分析，我们进一步优化并获得的抗甲型流感病毒活性更好的化合物11q。化合物11q具有良好的口服生物利用度，通过口服给药在小鼠体内对甲型流感病毒具有很强的抑制活性。这些结果表明11q有望为开发新型抗流感病毒药物提供先导化合物。我们通过时间梯度递加实验、报告基因实验及微量热泳动实验确认了该类化合物通过靶向甲型流感病毒RNA依赖性RNA聚合酶（RdRp）的PB1亚基而发挥抗病毒活性。此外，化合物11q对大多数甲型流感病毒和乙型流感病毒均表现出很强抑制活性。通过该项目的实施，我们已发表SCI论文6篇，获批发明专利6项，培养博士生4人，硕士研究生5人。