STRUCTURE-GUIDED RECEPTOR/INHIBITOR TRIMERIZATION AND RELATED STRATEGIES AGAINST CORONAVIRUSES

结构引导的受体/抑制剂三聚化及相关抗冠状病毒策略

基本信息

批准号：
10671214
负责人：
Ailong Ke
金额：
$ 68.63万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-17 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10671214
关键词：
2019-nCoV ACE2 Aerosols Affinity Animal Model Avidity Binding COVID-19 COVID-19 pandemic Cell Culture Techniques Cells Cessation of life Coronavirus Coronavirus spike protein Data Disease Outbreaks Economics Effectiveness Future Geometry Hamsters Human Infection Knowledge Lead Mainstreaming Medical Mesocricetus auratus Middle East Respiratory Syndrome Middle East Respiratory Syndrome Coronavirus Modeling Molecular Molecular Conformation Mutation Nose Peptides Pharmaceutical Preparations Plant Roots Preclinical Testing Property Public Health Resistance SARS coronavirus SARS-CoV-2 infection SARS-CoV-2 transmission SARS-CoV-2 variant Safety Severe Acute Respiratory Syndrome Specificity Structure Testing Therapeutic Treatment Efficacy Viral Virus Virus Receptors Zoonoses base clinically relevant conformational conversion design follow-up global health human coronavirus improved in vivo inhibitor insight nanobodies nanomolar neutralizing antibody nonhuman primate novel prevent prophylactic protein function receptor receptor binding vaccine-induced antibodies

项目摘要

Project Summary The current SARS-CoV-2 pandemic poses an immediate and global public health threat. This is the third major zoonotic-born coronavirus outbreak in humans in the past twenty years, after SARS-CoV-1 and MERS-CoV. Because mainstream antiviral approaches such as vaccines and neutralizing antibodies have to be specifically developed for each virus, there has always been a lag in effective medical remedy, which resulted in grave human death toll and economic disruption. Since viruses rarely switch their receptor specificity, here we propose a therapeutic strategy that utilizes the trimeric receptor as a decoy to neutralize the virus. SARS-CoV- 1 and SARS-CoV-2 target the angiotensin-converting enzyme 2 (ACE2) to gain cell entry. The “tri-ACE2” decoys are designed in a structure-guided fashion to match the symmetry and geometry of the viral spike (S), which maximizes the binding affinity through the trimer avidity effect. Additional functional domains are then introduced into the tri-ACE2 platform to achieve better antiviral activity. We present strong preliminary data demonstrating the effectiveness of such designs. Tri-ACE2 decoys lock the viral spikes in a symmetric “3-UP” receptor binding domain (RBD) conformation, neutralizing various ACE2-tropic coronaviruses with nanomolar concentrations, ~100-fold better than monomeric ACE2. Replacing ACE2 with ab initial designed minibinders (miniature-ACE2s) further improved the IC50 to ~20 pM by enabling ultrafast S-binding. The advantage of tri- ACE2 over neutralizing antibodies lies in its potential broad-spectrum activity against all ACE2-tropic coronaviruses, and in its expected resistance against evader mutations in the viral receptor. Therefore, this line of inhibitors can potentially serve as an off-the-shelf therapeutic in future outbreaks caused by unknown ACE2- tropic coronaviruses. We propose to explore the full potential of the structure-guided receptor multimerization as a general antiviral strategy. These efforts will also generate new mechanistic insight about how the coronavirus spike protein functions in general. The specific aims include: 1) Produce potent and evader- resistant tri-ACE2 inhibitors against SARS-CoV-2; 2) Produce tri-miniatureACE2 with picomolar inhibitory activity and novel antiviral mechanism; 3) Determine the efficacy of tri-ACE2 against SARS-CoV-2 and related viruses in primary airway cell cultures and animal models; 4) Determine the prophylactic and therapeutic efficacy of tri-ACE2 based inhibitors against SARS-CoV-2 infection in a Golden Syrian hamster model. Upon the completion of this project, we expect that we will have developed safe, highly effective, and broad- spectrum anti-coronavirus drugs that can directly lead to trials in nonhuman primates or humans.

项目摘要当前的SARS-COV-2大流行构成了直接和全球的公共卫生威胁。这是第三大专业在SARS-COV-1和MERS-COV之后，过去二十年来人动物造成的冠状病毒爆发。因为必须专门为每种病毒开发的，一直有效的医疗疗法滞后，这导致了严重性人类死亡人数和经济破坏。由于病毒很少切换其接收器特异性，因此我们在这里提案一种治疗策略，利用三聚体受体作为中和病毒的诱饵。 sars-cov- 1和SARS-COV-2靶向血管紧张素转换酶2（ACE2）以获得细胞的进入。 “ tri-ace2” 诱饵以结构引导的设计，以匹配病毒尖峰的对称和几何形状，通过触发效应最大化结合亲和力。然后是其他功能域引入TRI-ACE2平台以获得更好的抗病毒活性。我们提供强大的初步数据证明此类设计的有效性。 Tri-ace2诱饵将病毒尖峰锁定在对称的“ 3-up”中受体结合结构域（RBD）构象，用纳摩尔中和各种ACE2-热带冠状病毒浓度，比单体ACE2好约100倍。用最初设计的小型插件代替ACE2 （微型ACE2S）通过启用超快的S结合，将IC50进一步提高到〜20 pm。三 - 优势 ACE2过度中和抗体在于其对所有ACE2-循环的潜在广谱活性冠状病毒及其对病毒受体中逃避突变的预期性。因此，这线在未知ACE2-引起的未来爆发中，抑制剂可能会成为一种现成的疗法热带冠状病毒。我们建议探索结构引导的接收器多聚化的全部潜力作为一般抗病毒策略。这些努力还将产生有关如何冠状病毒峰值蛋白通常功能。具体目的包括：1）产生潜力和逃避者 - 抗SARS-COV-2的抗性TRI-ACE2抑制剂； 2）用皮摩尔抑制剂产生三痛2 活性和新型抗病毒机制； 3）确定针对SARS-COV-2和相关的TRI-ACE2的效率初级气道细胞培养和动物模型中的病毒； 4）确定预防和治疗在黄金叙利亚仓鼠模型中，基于TRI-ACE2对SARS-COV-2感染的抑制剂的效率。之上该项目的完成，我们希望我们将发展安全，高效且广泛 - 可以直接导致非人类隐私或人类试验的频谱抗核心病毒药物。