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.

摘要 在之前由美国国立卫生研究院赞助的研究中，我们成功地测试了这样一个假设，即将结构和 促进异多聚体VHH中和剂(VNA)设计的机理信息 开发具有更强疗效和通用性的抗毒素。在这份续订提案中，我们将应用这些 测试我们的设计者VNA平台将快速响应新威胁的假设的发现 允许开发高度实用的下一代抗毒素和抗病毒产品，这些产品具有出色的 治疗中毒或病毒感染的效力，对广泛的自然病原体有效 变种。我们的研究将集中在两种病原体上，这两种病原体是当前可能受益于下一步的主要威胁- 世代疗法：肉毒神经毒素(BONT)和SARS-CoV-2。我们提出了两个具体目标， 将在整个五年的研究中同时进行。在目标1中，我们将开发一个小型池 抗毒素VNA，可抵御三种流行的BONT血清型(A、B和E)的所有亚型。BoNTs是 CDC第1级精选代理。然而，针对BoNTs的少数可用的抗毒素治疗主要来自于 大型动物多克隆抗血清，如马肉毒毒素抗毒素HBAT，它们患有多发性 制造和存储方面的挑战。我们的目标是测试该平台产生高度实用的VNA的能力 作为下一代BoNT抗毒素产品，可能以RNA纳米颗粒的形式提供，它改进了 当前HBAT抗毒素产品的效力和自然变异特异性，并对 潜在的新的BONT威胁。在目标2中，我们将开发一种单一的VNA抗病毒药物，它可以预防已知的 SARS-CoV-1和SARS-CoV-2的变种。SARS-CoV-2是正在进行的新冠肺炎大流行的病毒原因。 快速开发治疗方法的一个有希望的策略是开发SARS-CoV-2中和抗体， 尤其是针对Spike蛋白的抗体，用于预防性或被动免疫治疗。然而，小说 导致感染和传播增强的SARS-CoV-2变种已经出现，而且还有更多 预计会出现危险的变异。直接令人担忧的是部分逃脱的变体 通过当前基于抗体的治疗以及在接种疫苗或以前感染过新冠肺炎的患者中中和，领先 在这些变种高度流行的某些地区，疫苗效力下降。我们提出一种信使核糖核酸 基于抗病毒产品，一旦给药，就会诱导具有极高病毒中和能力的VNA表达 威力。VNA将包含多个共价连接的VHH，与Spike蛋白的保守表位结合。 这种方法将测试平台开发将免疫逃逸风险降至最低的产品的能力 通过对SARS-CoV-2和SARS-CoV-1临床毒株的进化和筛选。如果成功，这将是 技术平台在为各种疾病创造实用疗法方面可以有广泛的应用 新出现的和潜在的大流行性病毒感染、生物恐怖威胁因子和其他传染病。