Application of cdk2 kinase inhibitor in preparation of hepatitis B therapeutic drug
A kinase inhibitor, hepatitis B technology, applied in the field of drug research and development, can solve the problem of unclear regulation of host protein kinase phosphorylation, and achieve the effect of broad application prospects
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Embodiment 1
[0017] Example 1 Identification and screening of host protein kinases involved in regulating the phosphorylation of restriction factor SAMHD1 In order to identify and screen host protein kinases involved in the regulation of phosphorylation of restriction factor SAMHD1, the applicant specifically interfered with cyclin-dependent kinase (cyclin -dependent kinases, CDK1, CDK2 and CDK6) (interfering sequence as shown in Table 2 in embodiment 2) expression, analyze the change of virus replication level and the change of SAMHD1 phosphorylation level (analysis method and the analysis in embodiment 2 The method is the same): the results show that (such as figure 1 (shown) After interfering with the expression of CDK2 kinase, it can obviously lead to a decrease in the phosphorylation of the restriction factor SAMHD1 and a decrease in the level of HBV replication, indicating that CDK2 kinase is involved in the phosphorylation of the restriction factor SAMHD1, and CDK2 / SAMHD1 can be used...
Embodiment 2
[0019] On the basis of the applicant's research in Example 1: it was found that CDK2 kinase inhibitors can inhibit HBV replication by inhibiting the phosphorylation of the host-restricted factor SAMHD1; in the HBV replication transfection model and based on NTCP-HepG2.0 In the infection model, small molecular compounds that exert antiviral effects were screened from existing CDK2 kinase inhibitors. The experimental procedure is:
[0020] (1): Firstly, the toxicity of CDK2 kinase inhibitors from Selleck Company, Santa Company and Merckmillipore Company to hepatocytes was detected by MTT method in the cell line HepAD38 cells with stable replication of HBV;
[0021] (2) In the cell line HepAD38 cells of HBV stably replicated in co-transfected SAMHD1, detect the effect of the above-mentioned CDK2 kinase inhibitor on the inhibition of virus replication and the phosphorylation of host restriction factor SAMHD1;
[0022] (3) On the HBV infection model based on NTCP-HepG2.0, verify t...
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