Pan-Influenza virus inhibitors: Small molecules that disrupt host ADAR1-viral NS1 interactions

 描述(申请人提供)：甲型流感病毒对人类健康构成严重威胁，导致季节性流行病和偶尔的大流行，导致全球范围内的严重发病率和死亡率。预防季节性流感病毒最有效的方法是接种疫苗；然而，疫苗对新型流感病毒无效，因为季节性疫苗很少或根本没有交叉保护作用。由于生产足够数量的流感疫苗至少需要6个月，而建立群体免疫是一个漫长而复杂的过程，所有年龄段的人都非常容易感染新的流感病毒株。为了克服现有流感疫苗的局限性，作为流感病毒感染的替代疗法，小分子抑制剂仍然是至关重要的需要。不幸的是，耐药性使现有疗法的有效性大大降低，仍然需要新的抗病毒药物来对抗未来的流行病和大流行。我们最近发现作用于双链RNA(ADAR1)的腺苷脱氨酶是多种流感病毒复制的关键宿主因子。重要的是，我们已经确定流感病毒的非结构蛋白1(NS1)与ADAR1相互作用并增强其RNA编辑活性，这是病毒有效复制和维持后代适应性所必需的。我们的数据表明，在缺乏ADAR1的细胞或表达催化失活的ADAR1的细胞中，流感病毒的复制减少了10-1000倍；此外，在缺乏ADAR1的情况下传代的病毒无法获得耐药性突变。我们已经建立了一种基于荧光素酶的检测ADAR1脱氨酶活性的方法，并在高通量筛选(HTS)系统中验证了它的稳健性。我们将筛选干扰ADAR1-NS1相互作用和/或直接抑制ADAR1脱氨酶活性的小分子抑制剂，通过合成孔径雷达研究提高已识别化合物的效力和选择性，在携带流感病毒的小鼠身上进行临床前评估研究，并开发适合进一步临床前测试的先进先导化合物。

 DESCRIPTION (provided by applicant): Influenza A viruses pose a serious threat to human health, causing seasonal epidemics and occasional pandemics that result in significant morbidity and mortality worldwide. The most effective method of protection against seasonal influenza virus is vaccination; however, vaccines are ineffective against novel influenza viruses, as there is little or no cross protection afforded by seasonal vaccines. Because production of sufficient quantities of influenza vaccine requires at least 6 months, and establishment of herd immunity is a lengthy and complicated process, people of all ages are highly vulnerable to novel influenza virus strains. To overcome the limitations of the available influenza vaccines, small molecule inhibitors are still a vital need as an alternative treatment to influenza virus infection Unfortunately, resistance has rendered existing therapeutics much less effective, and the need remains for new antiviral drugs to combat future epidemics and pandemics. We have recently identified Adenosine Deaminase Acting on double stranded RNA (ADAR1) as a host factor critical to the replication of multiple strains of influenza viruses. Importantly, we have determind that the non- structural protein 1 (NS1) of influenza virus interacts with ADAR1 and enhances its RNA editing activity, which is necessary for efficient viral replication and maintenance of progeny fitness. Our data demonstrates that influenza virus replication is reduced 10-1000 fold in cells lacking ADAR1 or cells expressing a catalytically inactive ADAR1; furthermore, viruses passaged in the absence of ADAR1 fail to acquire resistance mutations. We have developed a luciferase-based assay to measure the deaminase activity of ADAR1 and have validated its robustness in the high throughput screening (HTS) system. We will screen for small molecule inhibitors that disrupt ADAR1-NS1 interactions and/or directly inhibit ADAR1 deaminase activity, improve the potency and selectivity of identified compounds by SAR studies, perform preclinical evaluation studies in mice with influenza viruses, and develop advanced lead compounds suitable for further preclinical testing.