Project 3: Pandemic Virus Protease Inhibitors

项目3--大流行病毒蛋白酶抑制剂 摘要 病毒蛋白酶是高度优先的药物靶点，因为它们在病毒复制中具有基本功能，而且 明确的可药性证据，艾滋病毒和丙型肝炎药物是主要的成功故事。这个项目是 组织的目标是开发新型冠状病毒和黄病毒蛋白水解酶的化学抑制剂。 具体地说，SARS-CoV-2有两种蛋白酶，类木瓜蛋白酶(PLPRO)和主蛋白酶(MPRO)， 分别催化3次和11次病毒多蛋白切割事件。SARS-CoV-2 MPRO是一个有吸引力的药物靶点 由于在病毒感染的最早阶段有大量的基本裂解，而且它已经 多个可用药表面，包括半胱氨酸可共价加成的活性部位。它还分享了 冠状病毒MPRO相关蛋白的机制和结构特征，提示有可能 开发一种泛冠状病毒MPRO抑制剂，用于对抗当前的大流行病毒和未来用于预防 未来冠状病毒人畜共患病事件。同样，黄病毒，如寨卡病毒，需要蛋白水解酶的功能 最早的病毒生命周期阶段。寨卡病毒NS2B-NS3蛋白酶也有多个可药物表面，并且 与相关的黄病毒蛋白水解酶的氨基酸和结构相似表明有可能获得更广泛的光谱 抑制力。作为初步研究，我们设计并优化了这些病毒蛋白水解酶的多种检测方法， 有助于丰富这些药物靶标的结构、生物物理和计算知识，以及 已获得多个系列候选小分子抑制剂。我们依次提出了两个具体目标 加快实现我们的中心目标。在目标1中，我们将利用我们的分析并与我们的 筛选(核心B)和结构/计算/病毒学核心(核心D-E)以识别额外的小分子 这两种酶的抑制剂，以最大限度地获得进一步开发的热门系列的机会。在……里面 目标2，候选命中将通过在直系第二和第三级进行测试来优先进行进一步开发 分析，并与我们的化学/DMPK和结构/计算核心(核心C-D)密切合作， 通过系统地设计和测试相关的商业和新型合成分子来阐述。完全没有 在发展阶段，将使用细胞分析对每个系列中具有代表性的热门歌曲进行定量评估 而且，与我们的病毒学核心(核心E)密切合作，也使用最好的可用系统进行了严格的测试 用于病毒复制和体内致病。这一高度集成和协作的 努力将多种新型先导抑制剂用于临床前和临床试验，希望与行业合作伙伴 很快就会加强结束新冠肺炎大流行并帮助中和所需的药物 未来的疫情。

Project 3 – Pandemic Virus Protease Inhibitors ABSTRACT Viral proteases are high-priority drug targets due to their essential functions in virus replication and also unambiguous evidence for druggability, with HIV and HCV drugs as major success stories. This project is organized with the goal of developing novel chemical inhibitors of coronavirus and flavivirus proteases. Specifically, SARS-CoV-2 has two proteases, Papain-Like Protease (PLPro) and Main Protease (MPro), that catalyze 3 and 11 viral polyprotein cleavage events, respectively. SARS-CoV-2 MPro is an attractive drug target due to the large number of essential cleavages at the earliest stages of viral infection and the fact that it has multiple druggable surfaces, including an active site cysteine amenable to covalent adduction. It also shares mechanistic and structural features with related coronavirus MPro proteins, which suggests that it may be possible to develop a pan-coronavirus MPro inhibitor for use against the current pandemic virus and for future use against future coronavirus zoonotic events. Similarly, flaviviruses such as Zika virus require protease function for the earliest viral life cycle stages. The Zika virus NS2B-NS3 protease also has multiple druggable surfaces, and amino acid and structural similarities with related flavivirus proteases suggest potential for broader spectrum inhibition. As preliminary studies, we have designed and optimized multiple assays for these viral proteases, contributed to the wealth of structural, biophysical, and computational knowledge of these drug targets, and already obtained multiple series of candidate small molecule inhibitors. We propose two specific aims in order to expedite the achievement of our central goal. In Aim 1, we will leverage our assays and work closely with our screening (Core B) and structural/computational/virology cores (Cores D-E) to identify additional small molecule inhibitors of these two enzymes in order to maximize chances of obtaining hit series for further development. In Aim 2, candidate hits will be prioritized for further development by testing in orthologous secondary and tertiary assays and, in close collaboration with our chemistry/DMPK and structural/computational cores (Cores C-D), elaborated by systematically designing and testing related commercial and novel synthesized molecules. At all stages of development, representative hits from each series will be evaluated quantitatively using cellular assays and, in close collaboration with our virology core (Core E), also tested rigorously using the best available systems for virus replication and pathogenesis in vivo. The major deliverable from this highly integrated and collaborative effort will be multiple novel lead inhibitors for pre-clinical and clinical testing with industry partners with the hope of soon-fortifying the arsenal of drugs that will be required to end the COVID-19 pandemic and help neutralize future outbreaks.