Project 2: Nanobodies as Novel Entry Inhibitors of Pandemic Viruses

项目 2：纳米抗体作为大流行病毒的新进入抑制剂

• 批准号: 10522811
• 负责人: Lanying Du
• 金额: $ 377.42万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10522811
• 关键词: 2019-nCoV; Affinity; Animal Model; Antibodies; Antiviral Agents; Arenavirus; Binding; Biological Assay; Blood; Bolivian Hemorrhagic Fever Virus; COVID-19 pandemic; COVID-19 therapeutics; Cells; Chemistry; Clinical; Complex; Coronavirus; Cryoelectron Microscopy; Cultured Cells; Data; Development; Disease; Drug Kinetics; Drug Targeting; Drug or chemical Tissue Distribution; Drug usage; Ebola; Ebola virus; Engineering; Epitopes; Filovirus; Flavivirus; Future; Glycoproteins; Goals; Half-Life; Human; Immunize; In Vitro; Infection; Inhalators; Kidney; Lassa virus; Libraries; Lung; Mediating; Membrane; Membrane Fusion; Membrane Glycoproteins; Midwestern United States; Mission; Modeling; Molecular Weight; Mus; Names; Phage Display; Pharmaceutical Preparations; Preventive; Process; Production; RNA Viruses; Research; Safety; Series; Site; Solid; Specificity; Structure; Testing; Therapeutic; United States National Institutes of Health; Validation; Variant; Viral; Virus; Virus Diseases; Virus Receptors; X-Ray Crystallography; ZIKA; Zika Virus; aerosolized; base; chemical stability; cost; cost effective; drug candidate; improved; in vivo; inhibitor; nanobodies; novel; novel coronavirus; pandemic disease; pathogenic virus; programs; rational design; receptor binding; response; screening; side effect; small molecule; small molecule inhibitor; structural biology; success; therapeutic target; therapeutically effective; viral entry inhibitor; virology

Project 2 – Nanobodies as Novel Entry Inhibitors of Pandemic Viruses Summary Nanobodies (Nbs) are single domain antibodies derived from camelid heavy chain-only antibodies. Due to their small size, Nbs have many advantages over conventional antibodies as human therapeutics, such as their access to cryptic sites on targets, ease of production, superior pharmacokinetics, and strong physical and chemical stabilities. Nbs also have many advantages over small molecule drugs as human therapeutics, such as their high on-target specificity and low side effects. Several Nb drugs have been clinically approved to treat diseases in humans, confirming the safety and efficacy of Nbs as human therapeutics. A novel coronavirus (CoV) SARS-CoV-2 has caused the global COVID-19 pandemic. The fast emergence of many SARS-CoV-2 variants calls for urgent need of potent and broad-spectrum anti-COVID-19 therapeutics. Compared to small- molecule antiviral drugs, Nbs are particularly powerful in battling SARS-CoV-2 variants because they can be quickly adapted to new viral variants through phage display. Moreover, other pathogenic viruses also demonstrate pandemic potential, such as Ebola filovirus (EBOV), Lassa arenavirus (LASV)/Machupo arenavirus (MACV), and Zika flavivirus (ZIKV). These RNA viruses all contain a surface glycoprotein that mediates virus entry into host cells, thus the viral glycoprotein serves as a key therapeutic target. The current AViDD RFA program specifically includes Nb antiviral drugs as one of its missions. Therefore, Project 2 proposes to develop highly effective Nbs that target conserved epitopes of viral glycoproteins as novel inhibitors to block viral entry of these pandemic viruses. In our prior studies, we have developed several potent anti-CoV Nbs, including a series of anti-SARS-CoV-2 Nb candidate drugs named Nanosota-1. We have established camelid Nb phage display library platforms for Nb screening. We hypothesize that Nbs with high potency, good stability, low production costs, minimal side effects, superior pharmacokinetics, and broad antiviral spectrum can be developed as novel antiviral therapeutics. This project has three specific aims. In Aim 1, we will screen for antiviral Nbs using naïve or immunized Nb phage display libraries. We will also use in vitro affinity maturation to optimize the target-binding affinity of discovered Nbs. In Aim 2, based on structural information, we will engineer Nbs to further improve their target-binding affinity and antiviral potency. We will also improve Nb’s pharmacokinetics and minimize their side effects. In Aim 3, we will test and validate Nb candidate drugs in animal models against viral infections. This project is built upon a strong research team with complementary expertise in coronaviruses, filoviruses, arenaviruses, and flaviviruses, solid preliminary data, well-established platforms, and full support from the administration, chemistry, structural biology, and virology cores (Cores A and C-E). The overall goals of Project 2 are: (i) to discover Nb therapeutics against SARS- CoV-2 and its variants, helping ending the COVID-19 pandemic and (ii) to establish Nbs as potent, safe, and cost-effective therapeutics against other pathogenic viruses with pandemic potential.

项目2 - 作为大流行病毒的新型进入抑制剂的纳米体系 概括 纳米型（NBS）是源自骆驼重链抗体的单域抗体。由于 NBS小的尺寸比常规抗体具有许多优势，例如人类治疗 他们访问目标，易于生产，出色的药代动力学以及强大的身体和强大的物理和 化学系统。 NBS比小分子药物具有许多优势，例如人类治疗 由于它们的高目标特异性和低副作用。几种NB药物已被临床批准用于治疗 人类疾病，确认NBS作为人类治疗的安全性和效率。一种新颖的冠状病毒 （COV）SARS-COV-2引起了全球Covid-19-19。许多SARS-COV-2的快速出现 变体要求迫切需要有效和广泛的抗CoVID-19疗法。与小 分子抗病毒药物，NBS在战斗的SARS-COV-2变体中特别强大，因为它们可以是 通过噬菌体显示，迅速适应了新的病毒变体。此外，其他病毒病毒也 展示大流行潜力，例如埃博拉病毒（EBOV），LASSA AREAVIRUS（LASV）/MACHUPO Arenavirus（MACV）和Zika Flavivivirus（Zikv）。这些RNA病毒都含有表面糖蛋白 介导病毒进入宿主细胞，因此病毒糖蛋白是关键的治疗靶点。电流 Avidd RFA计划专门包括NB抗病毒药物作为其任务之一。因此，项目2 提出发展靶向病毒糖蛋白表位的高效NB的提议 阻止这些大流行病毒的病毒进入的抑制剂。在我们先前的研究中，我们开发了几个 潜在的抗COV NB，包括一系列名为Nanosota-1的抗SARS-COV-2 NB候选药物。我们有 建立的Camelid NB噬菌体展示库平台用于NB筛选。我们假设NB高 效力，良好的稳定性，低生产成本，最小的副作用，出色的药代动力学和广泛 抗病毒光谱可以作为新型抗病毒疗法发展。该项目具有三个特定的目标。在 AIM 1，我们将使用幼稚或免疫的NB噬菌体显示库筛选抗病毒NB。我们还将使用 体外亲和力成熟以优化发现的NB的目标结合亲和力。在AIM 2中，基于结构 信息，我们将设计NBS，以进一步改善其目标结合亲和力和抗病毒效力。我们将 还可以改善NB的药代动力学并最大程度地减少其副作用。在AIM 3中，我们将测试和验证NB 针对病毒感染的动物模型中的候选药物。该项目建立在一个强大的研究团队的基础上 完全专业的冠状病毒，丝状病毒，体育症病毒和黄病毒，可靠的初步数据， 良好的平台以及政府，化学，结构生物学和病毒学的全力支持 内核（核A和C-E）。项目2的总体目标是：（i）发现针对SARS的NB疗法 - COV-2及其变体，帮助结束Covid-19-19的大流行，（ii）将NBS建立为潜在，安全和 针对具有大流行潜力的其他病原病毒的具有成本效益的治疗。