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

 描述（由申请方提供）：甲型流感病毒对人类健康构成严重威胁，可引起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的最有效方法是接种疫苗;然而，疫苗对新型流感病毒无效，因为季节性疫苗几乎没有或没有提供交叉保护。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，因此所有年龄段的人都非常容易受到新型流感病毒株的影响。为了克服可用流感疫苗的局限性，小分子抑制剂仍然是至关重要的需要，作为流感病毒感染的替代治疗。不幸的是，耐药性使现有治疗剂的有效性大大降低，并且仍然需要新的抗病毒药物来对抗未来的流行病和大流行病。我们最近已经确定腺苷脱氨酶作用于双链RNA（ADAR 1）作为一种宿主因子的复制的多株流感病毒的关键。重要的是，我们已经确定流感病毒的非结构蛋白1（NS 1）与ADAR 1相互作用并增强其RNA编辑活性，这对于有效的病毒复制和后代适应性的维持是必需的。我们的数据表明，在缺乏ADAR 1的细胞或表达无催化活性的ADAR 1的细胞中，流感病毒复制减少10-1000倍;此外，在不存在ADAR 1的情况下传代的病毒未能获得抗性突变。我们已经开发了一种基于酶的测定来测量ADAR 1的脱氨酶活性，并验证了其在高通量筛选（HTS）系统中的稳健性。我们将筛选破坏ADAR 1-NS 1相互作用和/或直接抑制ADAR 1脱氨酶活性的小分子抑制剂，通过SAR研究提高已鉴定化合物的效力和选择性，在携带流感病毒的小鼠中进行临床前评价研究，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.