A versatile structure-based therapeutic platform for development of VHH-based antitoxin and antiviral agents

一个多功能的基于结构的治疗平台，用于开发基于 VHH 的抗毒素和抗病毒药物

• 批准号: 10560883
• 负责人: Rongsheng Jin
• 金额: $ 86.57万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-04 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560883
• 关键词: 2019-nCoV; 3-Dimensional; Affinity; Animal Disease Models; Animal Model; Animals; Antibodies; Antibody Therapy; Antiviral Agents; Area; Autoimmune; Binding; Biological Assay; Botulinum Toxins; Botulism; COVID-19 pandemic; COVID-19 patient; Cell Culture Techniques; Characteristics; Clinical; Clostridium difficile; Communicable Diseases; Complex; Dangerousness; Data; Development; Disease; Epitopes; Equus caballus; Evolution; Family; Funding; Future; Goals; High Prevalence; Human; Immune; Immune Sera; Infection; Intoxication; Intramuscular; Lead; Length; Life; Link; Malignant Neoplasms; Mediating; Messenger RNA; Modeling; Monoclonal Antibodies; Mus; Passive Immunotherapy; Pathology; Poisoning; Production; Property; Prophylactic treatment; Proteins; RNA; Research; Route; SARS coronavirus; SARS-CoV-2 variant; Serotyping; Specificity; Structure; Testing; Therapeutic; Therapeutic Agents; Therapeutic Effect; Therapeutic Monoclonal Antibodies; Therapeutic antibodies; Toxin; United States National Institutes of Health; Vaccinated; Variant; Viral; Virus; Virus Diseases; Work; antitoxin; clinically relevant; cost; design; disorder prevention; human disease; improved; in vivo; manufacture; microbial; nanobodies; nanoparticle; neutralizing antibody; next generation; novel; novel therapeutics; pandemic potential; pathogen; porcine model; prevent; product development; prophylactic; public health relevance; receptor binding; risk minimization; standard of care; technology platform; therapeutic development; transmission process; vaccine efficacy; variants of concern; virtual

ABSTRACT In previous NIH sponsored research we successfully tested the hypothesis that integrating structural and mechanistic information into heteromultimeric VHH-based neutralizing agent (VNA) design facilitated development of antitoxins with even greater efficacy and versatility. In this renewal proposal, we will apply these findings to test the hypothesis that our designer VNA platform, which is rapidly responsive to new threats, will permit development of highly practical, next-generation antitoxin and antiviral products that possess excellent potencies in treating intoxications or viral infections and are effective against a broad range of natural pathogen variants. Our research will focus on two pathogens that are major current threats which could benefit from next- generation therapeutics: botulinum neurotoxin (BoNT) and SARS-CoV-2. We propose two Specific Aims which will be underway simultaneously throughout the five years of research. In Aim 1, we will develop a small pool of antitoxin VNAs that protect against all subtypes of the three prevalent BoNT serotypes (A, B and E). BoNTs are CDC Tier 1 select agents. However, the few available antitoxin treatments against BoNTs primarily derive from large animal polyclonal antisera, such as the equine botulism antitoxin HBAT, which suffer from multiple manufacturing and storage challenges. Our goal is to test the platform’s ability to produce highly practical VNAs as a next-generation BoNT antitoxin product, likely delivered as RNA nanoparticles, which improves on the potencies and natural variant specificities of the current HBAT antitoxin product and is rapidly responsive to potential new BoNT threats. In Aim 2 we will develop a single VNA antiviral agent that protects against known variants of SARS-CoV-1 and SARS-CoV-2. SARS-CoV-2 is the viral cause of the ongoing COVID-19 pandemic. A promising strategy for rapid development of a therapy is development of SARS-CoV-2 neutralizing antibodies, especially antibodies targeting the spike protein, for prophylactic or passive immunotherapies. However, novel variants of SARS-CoV-2, which cause enhanced infection and transmission, have emerged, and more dangerous variants are expected to evolve. Of immediate concern are variants that partially escape neutralization by current Ab-based therapies and in vaccinated or previously-infected COVID-19 patients, leading to reduced vaccine efficacy in certain areas with a high prevalence of these variants. We propose an mRNA- based antiviral product that, once administered, elicits expression of a VNA with extremely high virus neutralizing potency. The VNA will contain multiple covalently linked VHHs binding to conserved epitopes of the spike protein. This approach will test the platform’s ability to develop a product that minimizes the risks of immune escape through evolution and selection of clinical strains of SARS-CoV-2 and SARS-CoV-1. If successful, this technology platform could have broad applications in creating practical therapeutics for a wide variety of emerging and potential pandemic viral infections, bioterror threat agents, and other infectious diseases.

抽象的 在以前的NIH赞助研究中，我们成功地检验了整合结构和整合的假设 基于异型VHH的机械信息基于中和剂（VNA）设计促进 开发具有更高效率和多功能性的抗毒素。在此续签建议中，我们将应用这些 研究结果是为了测试我们的设计师VNA平台迅速响应新威胁的假设，将 允许开发具有优秀的高度实用，下一代抗毒素和抗病毒产品 治疗肠毒性或病毒感染的能力，并且有效地抵抗广泛的自然病原体 变体。我们的研究将集中于两种病原体，这些病原体是当前主要威胁，这些威胁可能会受益于下一步的病原体 生成疗法：肉毒杆菌神经毒素（BONT）和SARS-COV-2。我们提出了两个具体目标 在整个研究的五年中，将在进行中。在AIM 1中，我们将开发一个小池 抗毒素VNA可预防三种普遍的BONT血清型（A，B和E）的所有亚型。 Bonts是 CDC Tier 1选择代理。但是，少数针对BONT的可用抗毒素治疗源自 大动物多克隆抗血清，例如马肉毒杆菌抗毒素HBAT，患有多个 制造和存储挑战。我们的目标是测试平台生产高度实用的VNA的能力 作为下一代BONT抗毒素产物，可能以RNA纳米颗粒的形式递送，可改善 当前HBAT抗毒素产品的电位和自然变体规范，对 潜在的新BONT威胁。在AIM 2中，我们将开发一个单一的VNA抗病毒剂，以防止已知 SARS-COV-1和SARS-COV-2的变体。 SARS-COV-2是正在进行的Covid-19-19大流行的病毒原因。 快速发展治疗的有前途的策略是SARS-COV-2中和抗体的发展， 尤其是针对峰值蛋白的抗体，用于预防性或被动免疫疗法。但是，新颖 SARS-COV-2的变体已经出现了增强的感染和传播，并且更多 危险的变体有望发展。直接关注的是部分逃脱的变体 通过当前基于AB的疗法以及预疫苗或先前感染的Covid-19患者中和，领先 为了降低这些变体患病率高的某些区域的疫苗效率。我们提出了mrna- 基于抗病毒的产物，一旦给药，就会引起具有极高病毒中和的VNA的表达 效力。 VNA将包含多个共价链接的VHHS与尖峰蛋白的表位结合。 这种方法将测试平台开发产品的能力，以最大程度地减少免疫逃生的风险 通过进化和选择SARS-COV-2和SARS-COV-1的临床菌株。如果成功，这 技术平台可能在创建各种各样的实用疗法方面具有广泛的应用 新兴和潜在的大流行病毒感染，生物侵蚀剂和其他传染病。