The limited therapeutic options and increasing drug-resistance call for next-generation influenza antivirals. Due to the essential function in viral replication and high sequence conservation among influenza viruses, influenza polymerase PA-PB1 protein-protein interaction becomes an attractive drug target. Here, we developed an in vitro split luciferase complementation-based assay to speed up screening of PA-PB1 interaction inhibitors. By screening 10,000 compounds, we identified two PA-PB1 interaction inhibitors, R160792 and R151785, with potent and broad-spectrum antiviral activity against a panel of influenza A and B viruses, including amantadine-, oseltamivir-, or dual resistant strains. Further mechanistic study reveals that R151785 inhibits PA nuclear localization, reduces the levels of viral RNAs and proteins, and inhibits viral replication at the intermediate stage, all of which are in line with its antiviral mechanism of action. Overall, we developed a robust high throughput-screening assay for screening broad-spectrum influenza antivirals targeting PA-PB1 interaction and identified R151785 as a promising antiviral drug candidate.

中文翻译：

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使用基于体外分裂荧光素酶补体的检测方法发现流感聚合酶PA-PB1相互作用抑制剂。

有限的治疗选择和日益增加的耐药性要求下一代流感抗病毒药。由于流感病毒在病毒复制和高序列保守性方面起着重要作用，因此流感聚合酶PA-PB1蛋白质与蛋白质的相互作用成为有吸引力的药物靶标。在这里，我们开发了一种基于体外分裂荧光素酶补体的检测方法，以加快PA-PB1相互作用抑制剂的筛选。通过筛选10,000种化合物，我们鉴定了两种PA-PB1相互作用抑制剂R160792和R151785，它们对一组甲型和B型流感病毒具有强效和广谱抗病毒活性，包括金刚烷胺，奥司他韦或双重耐药株。进一步的机理研究表明，R151785可以抑制PA核的定位，降低病毒RNA和蛋白质的水平，并在中间阶段抑制病毒复制，所有这些都与其抗病毒作用机制相符。总体而言，我们开发了一种强大的高通量筛选测定方法，用于筛选靶向PA-PB1相互作用的广谱流感抗病毒药，并将R151785确定为有前途的抗病毒药物候选物。