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

基本信息

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
项目类别：
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

项目摘要

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 是一种新型冠状病毒，是当前全球大流行的病原体，已导致超过 200 万人感染、125,000 人死亡，并造成严重的经济影响。目前尚无可用的预防、治疗或疫苗，护理仅是支持性的。此外，21 世纪还出现了多种致命的人类冠状病毒（SARS-CoV、MERS-CoV 和现在的 SARS-CoV-2）。迫切需要新的选择来应对这一流行病和未来不可避免的流行病。冠状病毒利用多个家族的包膜病毒共享的保守进入机制感染细胞，其中三聚体病毒刺突蛋白的两个区域（HR1和HR2）形成高度稳定的6螺旋束结构，将病毒和细胞膜并置以诱导膜融合。抑制这种 6 螺旋束的形成可以阻止病毒进入并预防感染。我们的实验室专注于创新的对映体筛选技术（镜像噬菌体展示）与结构引导设计相结合，生产新型合成 D 肽病毒进入抑制剂，包括 CPT31，我们的高效、广泛活性的 HIV-1 D - 已被 FDA 批准用于临床研究的肽抑制剂。我们和其他人对 SARS 的研究表明，这些抑制策略应该能够成功阻止当前的冠状病毒。SARS 是一种高度相关的冠状病毒，与 SARS-CoV-2 的刺突 HR1 具有 86% 的同一性（和 95% 的相似性）。 D-肽（由镜像D-氨基酸组成的肽）不能被体内的蛋白酶消化，因此具有显着的治疗优势，包括延长半衰期、降低剂量、降低免疫原性（不被MHC呈递消化），以及在呼吸道等富含蛋白酶的环境中的耐久性。在这项为期一年的资助申请中，我们将首先设计和合成基于 SARS-CoV-2 三聚体 HR1 的肽。这些肽准确地模仿 HR1，因为它在进入病毒时出现在病毒上，因此是宝贵的药物筛选目标。然后，我们将使用这些模拟物作为镜像噬菌体展示的靶标，以识别 SARS-CoV-2 的 D 肽抑制剂，以阻断它和未来的 SARS 相关冠状病毒。