Project 3: Pandemic Virus Protease Inhibitors

项目3 -大流行病毒蛋白酶抑制剂 摘要 病毒蛋白酶是高优先级的药物靶标，这是由于它们在病毒复制中的基本功能， 明确的证据表明药物的可药用性，艾滋病毒和丙型肝炎病毒药物是主要的成功案例。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酶的化学抑制剂。 具体来说，SARS-CoV-2有两种蛋白酶，木瓜蛋白酶样蛋白酶（PLPro）和主蛋白酶（MPro）， 分别催化3个和11个病毒多蛋白裂解事件。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必需裂解， 多个可药用表面，包括一个活性位点半胱氨酸，适合共价加合。它还分享 与相关冠状病毒MPro蛋白的机制和结构特征，这表明它可能是可能的， 开发一种泛冠状病毒MPro抑制剂，用于对抗目前的大流行病毒，并用于未来对抗 未来冠状病毒人畜共患病事件。类似地，黄病毒如寨卡病毒需要蛋白酶功能来进行免疫。 最早的病毒生命周期阶段。寨卡病毒NS 2B-NS 3蛋白酶还具有多个可药物化表面， 与相关黄病毒蛋白酶的氨基酸和结构相似性表明具有更广谱的潜力 抑制作用作为初步研究，我们已经设计并优化了这些病毒蛋白酶的多种检测方法， 为这些药物靶点的结构、生物物理和计算知识的丰富做出了贡献， 已经获得了多个系列的候选小分子抑制剂。我们提出两个具体目标， 加快实现我们的中心目标。在目标1中，我们将利用我们的检测方法，并与我们的 筛选（核心B）和结构/计算/病毒学核心（核心D-E），以鉴定其他小分子 这两种酶的抑制剂，以最大限度地获得命中系列的机会，用于进一步发展。在 目标2：通过在中学和高等教育中进行测试， 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。根本 在开发阶段，将使用细胞试验定量评价每个系列的代表性命中 并与我们的病毒学核心（核心E）密切合作，使用最佳可用系统进行严格测试 用于病毒在体内的复制和致病。这种高度集成和协作的主要交付成果是 将努力与行业合作伙伴一起进行临床前和临床试验的多种新型铅抑制剂， 即将加强结束COVID-19大流行所需的药物库， 未来的爆发。

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.