Novel Peptide Fusion Inhibitors for the Treatment of COVID-19

用于治疗 COVID-19 的新型肽融合抑制剂

基本信息

批准号：
10379832
负责人：
ROBERT TARRAN
金额：
$ 30万
依托单位：
ELDEC PHARMACEUTICALS, INC.
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：
2022-08-23 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10379832
关键词：
2019-nCoV ACE2 Address Aerosols Affinity Angiotensin Receptor Animal Model Antiviral Agents Appearance Back Binding Binding Proteins Biological Availability Bronchoalveolar Lavage COVID-19 COVID-19 therapeutics COVID-19 treatment Cell membrane Cell physiology Cell surface Cells Chemicals Chemistry Collaborations Complex Consensus Sequence Coronavirus Crystallography Disease Doctor of Philosophy Effectiveness Environment Enzymes Epithelial Epithelial Cells Event Formulation Glosso-Sterandryl Glycoproteins Goals Guidelines HIV Head Human Immunology In Vitro Infection Infection prevention Inflammation Inhalation Inhalators Institutes Lead Leukocyte Elastase Lung Measures Medicine Membrane Fusion Microbiology Modeling Molecular Conformation Mus Nebulizer Pathogenicity Pathway interactions Patients Pattern Peptide Hydrolases Peptides Persons Pharmacologic Substance Pharmacology Phase Pneumonia Powder dose form Predisposition Primary Infection Process Proteins Proteolytic Processing RHO Effector Domain Recombinants Reproducibility Research Resistance Respiratory Tract Infections Risk Role SARS coronavirus Safety Severe Acute Respiratory Syndrome Severity of illness Smoke Smoker Surface Techniques Testing Therapeutic Time Tobacco smoke Transgenic Mice Variant Vesicular stomatitis Indiana virus Viral Virus Virus Diseases Work airway epithelium analog antiviral drug development base bronchial epithelium coronavirus antiviral coronavirus disease cytotoxicity effective therapy exposure to cigarette smoke immunogenicity improved in vivo inhibitor lead optimization neutrophil non-smoker novel novel coronavirus pandemic disease particle peptidomimetics prevent professor receptor receptor binding uptake

项目摘要

PROJECT SUMMARY COVID-19 is caused by inhalation of the latest coronavirus (CoV) SARS-CoV-2 into the lungs, and airway epithelia are particularly susceptible to uptake this virus. Extensive evidence indicates that angiotensin converting enzyme 2 (ACE2) binds to the S1 subunit of the SARS-CoV-2 Spike protein (S1), triggering selective proteolytic cleavage that liberates the S2 subunit. S2 undergoes extensive conformational changes to form a 6- helix bundle (6-HB) between Heptad Repeat (HR)-1 and HR-2 domains of S2, which ultimately results in the fusion of the viral particle with the cell membrane and subsequent viral entry. Based on the mechanism of viral entry, and supported by crystallography studies of the ACE2•S1 interface and the 6-HB complex of S2, enormous efforts are currently under way to develop peptide-based therapeutics to target both events: the interaction of SARS-CoV-2 Spike with ACE2 receptor, and the fusion of the viral particle to the cell membrane. We have discovered that exposure of well-differentiated, primary airway epithelial cultures to tobacco smoke for extended periods of time enhances ACE2 activity and increases binding of recombinant S1, which might explain the increased susceptibility of smokers to COVID-19. The Receptor Binding Domain (RBD) in S1 is part of a highly mutable region, as revealed by the appearance of multiple highly infectious SARS-CoV-2 variants in late 2020; thus, targeting this region might not be ideal for antiviral development. In contrast, the HR regions of the S2 subunit and the interaction mode of HR-1 and HR-2 domains within the 6-HB complex are highly conserved among various CoVs, which makes it an optimal target to develop broad-spectrum antivirals. EK1 is a peptide that. The goal of this application is to develop novel peptides that target the HR1 domain of the S2 subunit to inhibit membrane fusion and pseudovirus infection of SARS-CoV-2 as well as several other CoVs. These peptides should serve as broad-spectrum CoV antivirals for the treatment of COVID-19 and subsequent COVIDs. We propose to evaluate the proteolytic stability of several peptides in the hostile environment of the lung, as the main entry way of SARS-CoV-2, including stapled and N-capped peptides with enhanced helical constraint. We will measure the proteolytic stability of the peptides ex vivo using human lung secretions obtained from smokers and non-smokers. We will use primary airway epithelial cells to interrogate the ability of the peptides to inhibit fusion and SARS-CoV-2 pseudovirus infection to healthy and smoke-exposed airway cultures. The efficacy of these peptides will be ultimately evaluated in animal models. This study will address the feasibility of helical mimics to inhibit viral fusion and suppress viral entry into airway epithelia as a novel effective treatment against COVID-19.

项目概要 COVID-19 是由将最新的冠状病毒 (CoV) SARS-CoV-2 吸入肺部和气道引起的大量证据表明，上皮细胞特别容易吸收这种病毒。转换酶 2 (ACE2) 与 SARS-CoV-2 刺突蛋白 (S1) 的 S1 亚基结合，触发选择性释放 S2 亚基的蛋白水解裂解会经历广泛的构象变化，形成 6-。 S2 的七肽重复序列 (HR)-1 和 HR-2 结构域之间的螺旋束 (6-HB)，最终导致基于病毒的机制，病毒颗粒与细胞膜融合以及随后的病毒进入。条目，并得到 ACE2·S1 界面和 S2 的 6-HB 复合物的晶体学研究的支持，巨大的目前正在努力开发基于肽的疗法来针对这两个事件： SARS-CoV-2 与 ACE2 受体的刺突，以及病毒颗粒与细胞膜的融合。发现分化良好的初级气道上皮培养物长时间暴露在烟草烟雾中一段时间后会增强 ACE2 活性并增加重组 S1 的结合，这可能解释了 S1 中的受体结合域 (RBD) 是吸烟者对 COVID-19 的易感性增加的一部分。 2020 年底出现的多种高传染性 SARS-CoV-2 变体揭示了可变区域；因此，针对该区域可能不是抗病毒开发的理想选择，相反，S2 的 HR 区域。 6-HB复合物中HR-1和HR-2结构域的亚基和相互作用模式高度保守 EK1是多种冠状病毒中的一种肽，这使其成为开发广谱抗病毒药物的最佳靶标。本申请的目标是开发针对 S2 亚基的 HR1 结构域的新型肽。抑制 SARS-CoV-2 以及其他几种 CoV 的膜融合和假病毒感染。肽应作为广谱 CoV 抗病毒药物，用于治疗 COVID-19 和随后的 COVID。我们建议评估几种肽在肺部恶劣环境中的蛋白水解稳定性，因为 SARS-CoV-2的主要进入途径，包括具有增强螺旋约束的钉合肽和N封端肽。将使用从吸烟者获得的人肺分泌物来测量肽的离体蛋白水解稳定性我们将使用初级气道上皮细胞来研究肽的抑制能力。融合和 SARS-CoV-2 假病毒感染对健康和暴露于烟雾的气道培养物的功效。这些肽最终将在动物模型中进行评估，这项研究将解决螺旋的可行性。模拟抑制病毒融合并抑制病毒进入气道上皮，作为一种新型有效治疗方法新冠肺炎。