Broad-spectrum antiviral GS-5734 to treat MERS-CoV and related emerging CoV

项目摘要 人畜共患病毒，如丝状病毒和冠状病毒（CoV），对全球环境构成持续和日益增长的威胁。 公共卫生，因为它们不可预测地出现，造成毁灭性的大流行病爆发。在 21世纪世纪，严重急性呼吸综合征冠状病毒（SARS-CoV）和中东呼吸道 MERS冠状病毒（MERS-CoV）出现于人畜共患病毒库， 人类中东呼吸综合征冠状病毒在中东的骆驼中流行，在人类中不断出现新的感染。 虽然SARS-CoV目前还不是一种威胁，但已经从一些人中分离出了几种“大流行前”的SARS样CoV。 蝙蝠在人类细胞中高效复制，并对现有疗法产生抗药性。随着不可预测的 由于人类和野生动物生态的重叠，新型CoV出现在人类中的可能性很高， 很有可能。目前，没有批准的抗病毒疗法用于任何人类CoV感染。广谱 控制已知的人类和人畜共患CoV感染的CoV疗法将解决目前尚未解决的问题。 医疗需求，并可以应对未来的大流行事件。与吉利德科学公司合作， 证明核苷前药GS-5734在抑制多种人和 人畜共患病CoV体外和SARS-CoV体内。我们计划的主要目标是加速临床前 开发GS-5734并促进MERS CoV适应症的IND许可。为了彻底评估 抗病毒活性的广度并预测对未来新出现的CoV的疗效，我们还将评估疗效 针对代表全家族遗传多样性的一组CoV，包括大流行前的人畜共患病毒株 蓄势待发专注于高致病性MERS冠状病毒，我们独特的合作伙伴关系整合：i） 宏基因组学和重组病毒合成基因组恢复，ii）原代人肺细胞模型，iii） 尖端病毒学和生物化学，iv）人类疾病的稳健的鼠和灵长类动物模型，以及v） 艺术代谢和药代动力学分析。在目标1中，我们完善了药代动力学、药效学 GS-5734在各种原代人细胞中的疗效和代谢研究， 人和人畜共患病CoV的阵列，并通过评价在鼠和非人中的体内功效， MERS和SARS-CoV的灵长类模型。在目标2中，我们选择了对SARS-CoV和MERS的抗性- CoV，并确定抗性对病毒复制、适应性和治疗敏感性的影响。在Aim中 3.我们确定GS-5734的作用机制是否是直接影响病毒RNA复制的结果 和/或通过深度测序和单分子RNA荧光原位改变抗病毒免疫 载体或药物处理的感染细胞和小鼠的杂交。我们制定了广泛的发展战略- 可以扩展到威胁全球的多种新兴病毒病原体的光谱疗法 公共卫生

Project Summary Zoonotic viruses, like filoviruses and coronaviruses (CoV), represent a continuous and growing threat to global public health because they unpredictably emerge causing devastating outbreaks of pandemic disease. In the 21st century, severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) emerged from zoonotic pools of viruses, causing severe disease in humans. MERS-CoV is endemic in camels in the Middle East with continuous new infections in humans. Although SARS-CoV is not currently a threat, several “prepandemic” SARS-like CoVs have been isolated from bats that replicate efficiently in human cells and are resistant to existing therapies. With the unpredictable overlap of human and wild animal ecologies, the potential for novel CoV emergence into humans is highly probable. Currently, there are no approved antiviral therapies for any human CoV infection. Broad-spectrum CoV therapies that control known human and zoonotic CoV infections would address an immediate unmet medical need and could counter future pandemic episodes. In partnership with Gilead Sciences, we have demonstrated that the nucleoside prodrug, GS-5734, is highly efficacious in inhibiting multiple human and zoonotic CoV in vitro and SARS-CoV in vivo. The primary goal of our program is to accelerate the preclinical development of GS-5734 and promote IND licensure for the MERS-CoV indication. To thoroughly evaluate the breadth of antiviral activity and predict efficacy against future emerging CoV, we will also assess efficacy against a panel of CoV representative of family-wide genetic diversity, including prepandemic zoonotic strains poised for emergence. Focusing on the highly pathogenic MERS-CoV, our unique partnership integrates: i) metagenomics and recombinant virus synthetic genome recovery, ii) primary human lung cell models, iii) cutting edge virology and biochemistry, iv) robust murine and primate models of human disease and v) state of the art metabolic and pharmacokinetic analysis. In Aim 1, we refine the pharmacokinetics, pharmacodynamics and breadth of GS-5734 through efficacy and metabolism studies in various primary human cells with a diverse array of human and zoonotic CoV and through the evaluation of in vivo efficacy in murine and non-human primate models of MERS- and SARS-CoV. In Aim 2, we select for resistance against SARS-CoV and MERS- CoV, and determine the effect of resistance on virus replication, fitness and susceptibility to treatment. In Aim 3, we determine if the mechanism of action of GS-5734 is a result of direct effects on viral RNA replication and/or alteration of antiviral immunity via deep sequencing and single molecule RNA fluorescence in situ hybridization of vehicle or drug treated infected cells and mice. We articulate a development strategy for broad- spectrum therapeutics that could be extended to a multitude of emerging viral pathogens threatening global public health.