Engineering protease-resistant antiviral peptide inhibitors for SARS-CoV-2

设计针对 SARS-CoV-2 的蛋白酶抗性抗病毒肽抑制剂

基本信息

批准号：
10669579
负责人：
Anne Moscona
金额：
$ 71.87万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10669579
关键词：
2019-nCoV Amino Acids Animals Antiviral Agents Antiviral resistance Binding Biodistribution Biological Availability Biophysics C-terminal COVID-19 COVID-19 outbreak COVID-19 pandemic COVID-19 treatment Cell Culture Techniques Cell fusion Cell membrane Cell surface Cells Cholesterol Clinical Coronavirus Coronavirus Infections Couples Data Disease Dose Engineering Evolution Ferrets Foundations Future Generations Glycoproteins Goals HIV Half-Life Hamsters Human Immune response In Vitro Individual Infection Infection prevention Influenza Integration Host Factors Lipids Measles Measures Mediating Membrane Membrane Fusion Middle East Respiratory Syndrome Middle East Respiratory Syndrome Coronavirus Modeling Modification Molecular Molecular Conformation Parainfluenza Paramyxovirus Peptide Hydrolases Peptides Performance Periodicals Play Predisposition Prevention Process Prophylactic treatment Protein Engineering Proteins Proteolysis Regimen Resistance Role SARS coronavirus SARS-CoV-2 B.1.1.7 SARS-CoV-2 antiviral SARS-CoV-2 entry inhibitor SARS-CoV-2 infection SARS-CoV-2 inhibitor SARS-CoV-2 spike protein SARS-CoV-2 transmission Site Structure Therapeutic Tissues Toxic effect Translating Vaccines Vertebral column Viral Viral Physiology Virus Virus Diseases Virus-Cell Membrane Interaction Work airway epithelium anti-viral efficacy betacoronavirus clinical efficacy comparative efficacy design experimental study improved in vitro Assay in vivo inhibitor monolayer novel pathogenic virus peptide analog peptide drug prevent prophylactic protein aminoacid sequence prototype receptor receptor binding resistance mechanism uptake viral entry inhibitor viral transmission

项目摘要

No vaccines or treatments for SARS-CoV-2 are yet available. A simple prophylactic antiviral strategy would protect naïve individuals from infection now. In the future, when vaccines should be available, a prophylactic antiviral will be essential for individuals who do not mount a suitable immune response. Antivirals that target viral entry into the host cell have been proven effective against a wide range of viral diseases. The entry/fusion process for CoV (including SARS-CoV-2) is mediated by the viral envelope glycoprotein (S). Concerted action by the receptor-binding domain and the fusion domain is required for fusion. Upon viral attachment (and uptake in certain cases), large-scale conformational rearrangements occur in the fusion domain, driven by formation of a structure that couples protein refolding directly to membrane fusion. The formation of this structure can be targeted by fusion inhibitory peptides (C-terminal heptad repeat or HRC peptides) that prevent proper apposition of the HRC and HRN domains in S. We have found that conjugation of a lipid to an inhibitory peptide directs the peptide to cell membranes and increases antiviral efficacy. Analogous lipo-peptides prevent infection by several viruses (measles, Nipah, parainfluenza, influenza), and can be administered via the airway. Treatment is effective for some of these even several days after infection. In addition, we have shown that modifying the backbone of an HRC peptide via periodic replacement of α-amino acid residues with β- amino acid residues generates α/β-peptides that retain antiviral potency (toward HIV or parainfluenza) but are highly resistant to proteolysis. We recently generated an HRC lipopeptide that is effective against both SARS- CoV2 and MERS live viruses in vitro, blocks spread of SARS-CoV2 in human airway tissue, and inhibits transmission of SARS-CoV-2 between ferrets in direct contact. Here we propose to combine the lipid conjugation and backbone-modification strategies to generate potent inhibitors of SARS-CoV2 infection that display a long half-life in vivo. 1. Optimize the antiviral potency and bioavailability of SARS-CoV-2 HRC peptide fusion inhibitors via rational molecular engineering. Antiviral efficacy of α/β-lipopeptide candidates will be measured in quantitative in vitro assays, in authentic virus infection, and in a human airway model. 2. Evaluate the protection afforded by new backbone-modified α/β-lipopeptide fusion inhibitors against SARS-CoV-2 infection in hamsters. Analysis of in vivo biodistribution and toxicity of backbone modified S- CoV-2 α/β-lipopeptide fusion inhibitors and assessment of in vivo potency and resistance mechanisms will lay the foundation for a safe and effective SARS CoV-2 fusion inhibitor for coronavirus prevention and therapy.

目前还没有针对 SARS-CoV-2 的疫苗或治疗方法，只有简单的预防性抗病毒策略。现在，当疫苗可用时，可以保护幼稚的人免受感染。对于没有产生适当免疫反应的个体来说，抗病毒药物至关重要。病毒进入宿主细胞已被证明对多种病毒性疾病有效。冠状病毒（包括 SARS-CoV-2）的感染过程是由病毒包膜糖蛋白 (S) 介导的。受体结合结构域和融合结构域是病毒附着（和摄取）所必需的。在某些情况下），在融合域中发生大规模的构象重排，由形成将蛋白质重折叠直接耦合到膜融合的结构。以融合抑制肽（C 端七肽重复序列或 HRC 肽）为目标，可防止适当的融合 S 中 HRC 和 HRN 结构域的并置。我们发现脂质与抑制性物质的缀合肽将肽引导至细胞膜并增加类似脂肽的抗病毒功效。感染多种病毒（麻疹、尼帕病毒、副流感、流感），可通过此外，我们已经证明，即使在感染后几天，治疗也对其中一些有效。通过定期用 β- 氨基酸残基替换 α- 氨基酸残基来修饰 HRC 肽的主链氨基酸残基产生 α/β 肽，保留抗病毒效力（针对 HIV 或副流感），但我们最近产生了一种 HRC 脂肽，可有效对抗 SARS- CoV2 和 MERS 活病毒在体外，阻断 SARS-CoV2 在人体气道组织中的传播，并抑制 SARS-CoV-2 在直接接触的雪貂之间传播在此，我们建议将脂质结合起来。缀合和骨架修饰策略以产生 SARS-CoV2 感染的有效抑制剂显示出较长的体内半衰期。 1. 通过优化 SARS-CoV-2 HRC 肽融合抑制剂的抗病毒效力和生物利用度 α/β-脂肽候选物的抗病毒功效将在以下方面进行测量。定量体外测定、真实病毒感染和人体气道模型。 2. 评估新型主链修饰的 α/β-脂肽融合抑制剂针对仓鼠中 SARS-CoV-2 感染的骨架修饰 S- 的体内生物分布和毒性分析。 CoV-2 α/β-脂肽融合抑制剂以及体内效力和耐药机制的评估将奠定基础为安全有效的 SARS CoV-2 融合抑制剂预防冠状病毒奠定了基础治疗。