design and rapid production of a drug-screening target from the highly conserved HR1 region of the viral spike protein (S2)

从病毒刺突蛋白 (S2) 高度保守的 HR1 区域设计并快速生产药物筛选靶点

• 批准号: 10221150
• 负责人: Michael S Kay
• 金额: $ 44.94万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221150
• 关键词: 2019-nCoV; Affinity; Amino Acids; Antiviral Agents; Applications Grants; Bacteriophages; Binding; Biological Assay; Biophysics; Caring; Cells; Cellular Membrane; Cellular biology; Cessation of life; Chemicals; Cholesterol; Clinical Research; Collaborations; Complex; Coronavirus; Coronavirus spike protein; Dose; Drug Screening; Drug Targeting; Ebola; Elements; Environment; Family; Fluorescence Polarization; Future; Goals; HIV; HIV-1; Half-Life; Human; Image; Infection; Infection prevention; Institutes; Lead; Libraries; Ligation; Luciferases; Measures; Membrane; Membrane Fusion; Middle East Respiratory Syndrome Coronavirus; Modification; Peptide Hydrolases; Peptide Synthesis; Peptides; Phage Display; Phase; Preventive treatment; Process; Production; Proteins; Research; Resolution; Respiratory System; SARS coronavirus; Severe Acute Respiratory Syndrome; Solid; Structure; Surface Plasmon Resonance; Technology; Testing; Therapeutic; Therapeutic Index; Universities; Utah; Vaccines; Vesicular stomatitis Indiana virus; Viral; Virus; Work; X-Ray Crystallography; base; combat; cytotoxicity; design; drug discovery; economic impact; human coronavirus; immunogenicity; improved; inhibitor/antagonist; innovation; novel; novel coronavirus; pandemic disease; protein aminoacid sequence; scaffold; screening; structural biology; tool; trafficking; virus envelope

SARS-CoV-2, a novel coronavirus, is the causative agent of the current worldwide pandemic that has already led to over 2 million infections, 125,000 deaths, and severe economic impact. Currently no preventatives, treatments, or vaccines are available, and care is solely supportive. Additionally, the 21st century has seen the emergence of multiple lethal human coronaviruses (SARS-CoV, MERS-CoV, and now SARS- CoV-2). There is an urgent need for new options to combat this and inevitable future pandemics. Coronaviruses infect cells using a conserved entry mechanism shared by enveloped viruses across multiple families in which two regions of the trimeric viral spike protein (HR1 and HR2) form a highly stable 6- helix bundle structure, juxtaposing the viral and cellular membranes to induce membrane fusion. Inhibiting formation of this 6-helix bundle stops viral entry and prevents infection. Our lab specializes in an innovative enantiomeric screening technology (mirror-image phage display) in concert with structure-guided design, to produce novel, synthetic, D-peptide viral entry inhibitors, including CPT31, our highly potent, broadly active HIV-1 D-peptide inhibitor that has been cleared by the FDA for clinical studies. Research by us and others on SARS, a highly related coronavirus that shares 86% identity (and 95% similarity) to SARS-CoV-2 in the spike HR1, indicate that these inhibition strategies should successfully block the current coronavirus. D-peptides (peptides composed of mirror-image D-amino acids) cannot be digested by proteases in the body and, therefore, possess significant therapeutic advantages including extended half-life, lower dosing, reduced immunogenicity (not digested for MHC presentation), and durability in protease-rich environments such as the respiratory tract. In this one-year grant application, we will first design and synthesize SARS-CoV-2 trimeric HR1-based peptides. These peptides accurately mimic HR1 as it appears on the virus during entry and are therefore invaluable drug screening targets. We will then use these mimics as targets in mirror-image phage display to identify D-peptide inhibitors of SARS-CoV-2 that block it and future SARS-related coronaviruses.

SARS-COV-2是一种新颖的冠状病毒，是当前全球大流行的原因，已经导致超过200万感染，125,000人死亡和严重的经济影响。目前没有预防剂，治疗或疫苗可用，并且护理仅提供支持。此外，21世纪已经看到了多种致命的人类冠状病毒（SARS-COV，MERS-COV，现在是SARS-COV-2）的出现。迫切需要新的选择来打击这一和不可避免的未来大流行。 冠状病毒使用包裹病毒共享的保守进入机制感染细胞，在多个家族中，三聚体病毒尖峰蛋白（HR1和HR2）的两个区域形成了高度稳定的6-螺旋束结构，并并置，使病毒和细胞膜并置，以诱导膜融合。抑制此6螺旋束的形成可阻止病毒进入并防止感染。我们的实验室专门从事创新的对映体筛选技术（镜像图像噬菌体显示），以结构引导的设计共同生产新颖的，合成的，D肽病毒入口抑制剂，包括CPT31，包括我们高度有效的，广泛活跃的HIV-1 D肽抑制剂，该抑制剂已被FDA清除，该抑制剂已被FDA清除。我们和其他人在SARS上进行的研究是一种高度相关的冠状病毒，在SPIKE HR1中与SARS-COV-2具有86％的身份（和95％的相似性），表明这些抑制策略应成功阻止当前的冠状病毒。 D肽（由镜像D-氨基酸组成的肽）不能被体内蛋白酶消化，因此具有显着的治疗优势，包括延长的半寿命，较低的给药，降低的免疫原性（未在MHC表现中消化），以及诸如蛋白酶富含蛋白酶的环境中的持久性，例如蛋白酶，例如呼吸道。 在此为期一年的赠款应用中，我们将首先设计并合成基于HR1的SARS-COV-2三聚体。这些肽可以准确地模仿入口期间病毒上出现的HR1，因此是宝贵的药物筛查靶标。然后，我们将这些模拟物用作镜像噬菌体显示器中的目标，以识别阻断其和将来与SARS相关的冠状病毒的SARS-COV-2的D肽抑制剂。