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

• 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
• 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

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.

SARS-COV-2尚无疫苗或治疗方法。一个简单的预防性抗病毒策略将 现在保护幼稚的个体免受感染。将来，当应提供疫苗时，预防性 抗病毒对于不进行适当免疫反应的个体至关重要。靶向的抗病毒药 病毒进入宿主细胞已被证明有效地针对多种病毒疾病有效。条目/融合 COV的过程（包括SARS-COV-2）是由病毒包膜糖蛋白介导的。一致行动 融合需要由接收器结合域和融合域。病毒依恋（和吸收） 在某些情况下），大规模构象重排发生在融合域中，由形成。 将蛋白质直接重质膜融合到膜融合的结构。这种结构的形成可以是 以融合抑制性辣椒（C-末端七型重复或HRC Pepperides）为目标，可防止适当 S.中HRC和HRN结构域的方法 肽将肽引导到细胞膜并提高抗病毒效率。类似的脂肽预防 多种病毒感染（麻疹，尼帕，副氟氟扎，影响），可以通过 呼吸道。即使在感染后几天，治疗对于其中一些也有效。此外，我们已经显示 通过定期替换α-氨基酸残基，可以改变HRC胡椒的主链 氨基酸保留会产生α/β肽，可保留抗病毒效力（朝向HIV或Parainfluenza），但为 对蛋白水解高度抗性。我们最近产生了一个HRC脂蛋白肽，该脂蛋白肽有效地针对这两个SARS- COV2和MERS在体外的活病毒，SARS-COV2在人气道组织中扩散并抑制 直接接触的雪貂之间的SARS-COV-2传输。在这里，我们建议将脂质结合在一起 共轭和骨干 - 修饰策略，以产生SARS-COV2感染的潜在抑制剂 在体内表现出长的半衰期。 1。通过通过SARS-COV-2 HRC PEPPER融合抑制剂的抗病毒效力和生物利用度 理性分子工程。候选α/β-磷酸肽的抗病毒效率将在 定量体外测定，正宗病毒感染和人类气道模型。 2。评估新的骨干形修饰的α/β-黎肽融合抑制剂提供的保护 仓鼠中的SARS-COV-2感染。分析主链修饰S-的体内生物分布和毒性 COV-2α/β-硫代肽融合抑制剂以及对体内效力和耐药机制的评估 安全有效的SARS COV-2融合抑制剂预防冠状病毒和 治疗。