Broad-spectrum therapeutics against SARS-CoV-2 3CL protease

项目摘要/摘要 新冠肺炎于2019年12月首次发现在湖北省武汉市，中国，导致了正在进行的2019年- 2020年大流行。新冠肺炎是由严重急性呼吸系统综合症冠状病毒2型引起的。 2)。该病的常见症状包括发烧、干咳、呼吸急促、腹泻和嗅觉丧失。 并发症可能包括肺炎、病毒性败血症和急性呼吸窘迫综合征。截至今天，其他 目前还没有批准的小分子药物用于治疗新冠肺炎和发现 一种有效的疫苗仍然不确定。我们的长期目标是开发抗病毒药物来治疗 新冠肺炎和人类冠状病毒感染一般。我们的中心假设是抑制SARS-CoV- 2多聚蛋白切割可在新冠肺炎进展为更多之前预防和早期治疗 严重的形式。我们将确定适合于开发的冠状病毒3C样蛋白酶(3CLpro)的纳米分子抑制剂 作为治疗新冠肺炎和其他冠状病毒感染的抗病毒药物。该提案针对的是 3CLPro，SARS-CoV-2多蛋白裂解和病毒复制的关键酶。我们的总体前提是 针对这种基本病毒酶的小分子抑制剂将抑制复制，因此具有 具有潜在的预防和治疗价值。因此，我们的主要目标是设计和开发 利用我们已建立和验证的药物靶向冠状病毒3CLPro的结构型小分子抑制剂 发现专业知识。在强劲的初步数据指导下，多蛋白切割的抑制假说将被 通过追求三个特定目标进行测试：目的1)通过开发抑制SARS-CoV-2多蛋白切割 3CLPRO的共价肽抑制剂(NSP5)；目的2)通过开发抑制SARS-CoV-2多蛋白切割的方法 非共价非肽类3CLPro抑制剂(NSP5)；以及目的3)确定共价和 非共价SARS-CoV-2 3CLPro抑制剂在金黄地鼠模型中的应用。在第一个目标下，铅化合物 3150及其类似物将在病毒和酶试验中检测SARS-CoV-2 3CLPro的抑制活性。一个 3150的水溶形式将在动物模型中进行评估。基于结构的药物设计方法 将被用来优化3150与SARS-CoV-2 3CLPro的晶体结构结合。在目标2下， 基于结构的虚拟筛选和杂化配基筛选方法以及药物化学将 用于制备和评价3CLPro的非共价非肽类抑制剂。在第三个目标下，排名第一 将对SARS-CoV-2 3CLPro共价和非共价抑制剂进行药代动力学和疗效测试 金黄仓鼠新冠肺炎模型。这项拟议研究的最终目标是推动一场反对新冠肺炎的运动 候选药物进入提交研究用新药(IND)申请阶段。总体而言，这一结果 该项目将产生重大的积极影响，因为它们为临床开发奠定了基础 新冠肺炎抗病毒治疗以及将一种有效和选择性的蛋白酶抑制剂与一种 核苷类似物(例如，瑞希韦)，如果需要，抗炎药物(例如，地塞米松或巴利西尼)。

PROJECT SUMMARY/ABSTRACT COVID-19 was first identified in December 2019 in Wuhan, Hubei province, China, resulting in the ongoing 2019- 2020 pandemic. COVID-19 is caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV- 2). Common symptoms of the disease include fever, dry cough, shortness of breath, diarrhea, and loss of smell. Complications may include pneumonia, viral sepsis, and acute respiratory distress syndrome. As of today, other than remdesivir, there is no approved small molecule drug for the treatment of COVID-19 and the discovery of an effective vaccine remains uncertain. Our long-term goal is to develop antiviral drugs for the treatment of COVID-19 and human coronavirus infections in general. Our central hypothesis is that inhibition of SARS-CoV- 2 polyprotein cleavage results in the prevention and early treatment of COVID-19 before it progresses to its more severe form. We will identify nanomolar inhibitors of the CoV 3C-like protease (3CLpro) suitable to be developed as antiviral agents for the treatment of COVID-19 and other coronavirus infections. The proposal targets the 3CLpro, a key enzyme for SARS-CoV-2 polyprotein cleavage and viral replication. Our overall premise is that small molecule inhibitors targeting this essential viral enzyme will inhibit replication, and therefore have the potential to be of both preventive and therapeutic value. Thus, our primary objective is to design and develop structure-based small-molecule inhibitors targeting coronavirus 3CLpro using our established and proven drug discovery expertise. Guided by strong preliminary data, the inhibition of polyprotein cleavage hypothesis will be tested by pursuing three specific aims: Aim 1) To inhibit SARS-CoV-2 polyprotein cleavage by developing covalent peptidic inhibitors of 3CLpro (nsp5).; Aim 2) To inhibit SARS-CoV-2 polyprotein cleavage by developing noncovalent nonpeptidic inhibitors of 3CLpro (nsp5).; and Aim 3) To determine the efficacy of covalent and noncovalent SARS-CoV-2 3CLpro inhibitors in a golden hamster model. Under the first aim, lead compound 3150 and its analogs will be tested in viral and enzyme assays for inhibitory activity of SARS-CoV-2 3CLpro. An aqueous soluble form of 3150 will be evaluated in the animal model. Structure-based drug design approaches will be employed to optimize 3150 for binding to the crystal structure of SARS-CoV-2 3CLpro. Under aim 2, Structure-based virtual screening and hybrid ligand screening approaches along with medicinal chemistry will be used to prepare and evaluate noncovalent nonpeptidic inhibitors of 3CLpro. Under the third aim, top-ranked SARS-CoV-2 3CLpro covalent and noncovalent inhibitors will be tested for pharmacokinetics and efficacy in a golden hamster COVID-19 model. The ultimate goal of the proposed studies is to advance an anti-COVID-19 drug candidate to the stage of filing an investigational new drug (IND) application. Overall, the results of this project will have a significant positive impact because they lay the groundwork for the clinical development of COVID-19 antiviral therapy and the potential to combine a potent and selective protease inhibitor with a nucleoside analog (e.g., remdesivir) and if needed anti-inflammatory drugs (e.g., dexamethasone or baricitinib).