甲型流感病毒的基因易于变异，对于靶向病毒蛋白的药物易于产生耐药性，严重威胁人类的生命健康。选择宿主细胞中对宿主无关紧要但对病毒复制至关重要的蛋白，是解决病毒耐药性问题的重要途径。人源CDC样激酶1(CLK1)作为流感病毒复制过程中关键蛋白，有望成为抗流感药物新靶点。本课题将在CLK1抑制剂筛选与发现、抗流感活性评价的基础上,开展如下研究：通过建立CLK1基因敲除细胞模型和现有的CLK1过表达细胞模型，研究CLK1对病毒转录剪接因子SR磷酸化水平的影响、SR磷酸化对pre-mRNA选择性剪接和病毒蛋白表达的影响；采用细胞免疫荧光等方法，考察CLK1抑制剂对SR磷酸化，进而对pre-mRNA选择性剪接和病毒蛋白的调控作用；通过体内外实验，证实CLK1抑制剂对病毒耐药毒株的药效。以阐明CLK1及其抑制剂调控甲型流感病毒复制的机制，确证CLK1抑制剂优势，为靶向CLK1抗流感新药研发奠定理论基础。

Since the genes of Influenza A viruses are easy to mutate, it can easily produce drug resistance against the drugs targeting viral proteins, and therefore influenza A viruses seriously threaten human life and health. Selecting the host protein which is temporarily dispensable for the host but crucial for virus replication is an important way to solve the problem of drug resistance. Human CDC like kinase 1 (CLK1) as a key protein of influenza virus replication process is expected to become a new target for anti influenza drugs. The program will base on the CLK1 inhibitors screening and discovery and their anti-influenza activity evaluation, and carry out the following research: Through the establishment of CLK1 gene knockout cell model and the existing CLK1 over expression mode, the influence of CLK1 on the phosphorylation level of viral transcription splicing factor SR, and the regulation of the phosphorylated SR on pre-mRNA expression of alternative splicing and viral proteins will be studied; Using cell immunofluorescence method, the effects of CLK1 inhibitors on the phosphorylation of SR, and therefore regulating pre-mRNA alternative splicing and viral protein will be investigated; The efficacy of CLK1 inhibitors against virus resistant strains will be confirmed. Finally, the mechanism of CLK1 and its inhibitors regulating influenza A virus replication will be elucidated, the advantages of CLK1 will be confirmed. This program will lay important theoretical foundation for anti-influenza drug targeting CLK1.