抗寨卡病毒药物筛选模型的建立及其应用

摘要

Objective To establish a high-throughput screening (HTS) model for anti-Zika virus (ZIKV) compounds screening. Methods Using the CCK-8 assay, the viability of ZIKV infected cells were measured to monitor the replication of ZIKV. After optimization and validation of the screening model, the assay was used to screen 32 000 microorganism fermentation samples for the search of anti-ZIKV agents. Results Biochemical experiments and the statistical analysis presented in this work demonstrate the qualification of the screening model with regard to sensitivity and reproducibility, and the Z' factor, CV value, S/B and S/N is 0.733, 5.52％, 20.21 and 14.18, respectively. Using this model, 102 positive samples were obtained, among which the antiviral activity of a positive fermentation sample (I07A-01164) was verified with inhibiting more than 50％ of virus replication by four times dilution. Conclusion An ideal model is established for the evaluation of ZIKV inhibitors.%目的 建立寨卡病毒抑制剂高通量筛选模型, 为筛选和评价抗寨卡病毒化合物提供技术手段.方法 利用CCK-8法测定寨卡病毒感染后细胞活性, 间接反应胞内病毒复制水平.进而对分析方法进行优化, 建立可定量病毒复制能力的药物筛选模型, 并利用统计学分析对模型的稳定性和灵敏性进行评价.应用该模型对32 000个微生物发酵液进行初步筛选, 验证该方法的可行性.结果 成功建立寨卡病毒抑制剂高通量筛选模型, 该筛选模型具有较好的稳定性和灵敏度, Z'因子为0.733, CV为5.52％, S/B为20.21, S/N为14.18.利用该模型筛选得到阳性样品102个, 对其中I07A-01164进行抗病毒活性验证, 测定其稀释4倍后可以抑制50％以上的病毒复制.结论 建立了抗寨卡病毒化合物的高通量筛选模型, 可用于寨卡病毒抑制剂的筛选和活性初步评价, 有助于抗寨卡病毒药物的发现.