Antibody immunotherapeutics for biodefense against Lassa virus

用于对抗拉沙病毒生物防御的抗体免疫疗法

• 批准号: 9211282
• 负责人: Robert F Garry
• 金额: $ 114.85万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-27 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9211282
• 关键词: Achievement; Africa; Antibodies; Area; Arenaviridae; Arenavirus; B-Lymphocyte Epitopes; Biological Assay; Biological Warfare; Biotechnology; Blocking Antibodies; Categories; Cavia; Cell Line; Cell membrane; Chemicals; Clinical Research; Clinical Trials; Clone Cells; Developed Countries; Developing Countries; Development; Diagnostic; Disease; Dose; Drug Kinetics; Ensure; Evaluation; Future; Generations; Goals; Human; IgG1; IgG4; Immunologist; Immunotherapeutic agent; In Vitro; Individual; Infection; Intravenous; Laboratories; Laboratory Research; Lassa Fever; Lassa virus; Lead; Life; Mammalian Cell; Medical; Modernization; Molecular Biology Techniques; National Institute of Allergy and Infectious Disease; Nigeria; Pharmaceutical Preparations; Phospholipids; Production; Productivity; Proteins; Public Health; Recombinant Proteins; Resources; Route; Scientist; Serum; Sierra Leone; Technology; Texas; Therapeutic; Therapeutic antibodies; Time; TimeLine; Toxicology; Treatment Efficacy; Universities; Virus Diseases; base; biodefense; biothreat; cell bank; combinatorial; cost; cross reactivity; efficacy study; good laboratory practice; human monoclonal antibodies; in vitro testing; in vivo; interest; intraperitoneal; large scale production; milligram; murine monoclonal antibody; neutralizing monoclonal antibodies; nonhuman primate; novel vaccines; point of care; preclinical development; prevent; programs; public health relevance; social; stable cell line; subcutaneous

DESCRIPTION (provided by applicant): This project will develop Immunotherapeutics for Biodefense against Lassa virus (LASV), an arenavirus that is the causative agent of Lassa fever (LF). LASV is a Biosafety Level 4 and NIAID Biodefense category A agent. LF is a major public health concern causing widespread loss of life and social disruption across West Africa. The goal of this proposal is to perform critical steps in the characterization and pre-clinical development of a new class of therapeutics to treat LF. This effort combines the expertise of virologists and immunologists at Tulane University and the University of Texas Medical Branch with scientists at Zalgen, a biotechnology company specialized in the development of immunotherapeutics, rapid diagnostics, and new vaccine platforms for emerging viral diseases with global impact. Studies proposed here will leverage our recent advances, including identification of lead candidate human monoclonal antibodies (huMAbs) that neutralize LASV infectivity and protect guinea pigs against fatal disease induced by LASV. Zalgen's exclusive technology for rapid development (<3 months) of high yield antibody expressing stable cell lines ensures that costs of producing therapeutic antibodies is minimized for primary Biodefense markets and secondary Public Health markets. In studies proposed in MILESTONE 1 we will generate multi-milligram quantities of IgG1 and IgG4 LASV huMAb subtypes in serum-free medium adapted mammalian cell transient expression platforms for purification to perform in vitro testing. We will also perform a panel of characterization assays with the huMAbs focusing on identification of a huMAb or combinations of huMAbs with broad LASV neutralization potential and low IC50. Additional studies will establish cross-reactivity profiles with related Ne and Old World arenaviral proteins and reactivity of LASV huMAbs with human cell membrane phospholipids. In MILESTONE 2 we will generate CHO or NS0 stable cell lines in chemically defined serum free medium and select high producing isolates. Limiting dilution cell cloning (LDCC) will be performed for isolation of stable, production grade CD-SFM-CHO or CD-SFM-NS0 clones. We also plan to generate Accession Cell Banks (ACB) for clonal cell lines of interest, and to convert to manufacturing with QA/QC to generate gram quantities of candidate huMAbs for in vivo studies. In MILESTONE 3 we will perform evaluation of pharmacokinetics of LASV huMAb subtypes in guinea pigs (GPs). We will also perform challenge - therapeutic dose finding studies with two lead candidate LASV huMAbs or huMAb combinations in GPs, following a single dose via optimal delivery route. Next, we will determine the therapeutic efficacy with LASV huMAbs or huMAb combinations in Good Laboratory Practice (GLP) GP studies throughout the course of a lethal infection timeline, via the optimally determined route of administration. Results of the GP studies will guide an evaluation of pharmacokinetics of two single LASV huMAbs (IgG1 or IgG4) or huMAb combinations in a GLP nonhuman primate (NHP) study, following a single intravenous, intraperitoneal, or subcutaneous dose. We will also perform therapeutic efficacy studies with LASV huMAbs or huMAb combinations in GLP NHP studies throughout the course of a lethal infection timeline, following two doses via optimal delivery route, at varying times post infection. This proposed effort will result in a new class of immunotherapeutics for LF.

描述(由申请人提供)：该项目将开发针对拉萨病毒(LASV)的生物防御的免疫疗法，LASV是拉沙热(LF)的病原体。LASV是生物安全4级和NIAID生物防御A类药物。该病是一个重大的公共卫生问题，在整个西非造成广泛的生命损失和社会混乱。这项提案的目标是在表征和临床前开发一类治疗LF的新疗法方面执行关键步骤。这项工作将杜兰大学和德克萨斯大学医学分校的病毒学家和免疫学家的专业知识与Zalgen的科学家结合在一起，Zalgen是一家生物技术公司，专门为具有全球影响的新出现的病毒疾病开发免疫疗法、快速诊断和新的疫苗平台。这里提出的研究将利用我们最新的进展，包括鉴定主要候选人类单抗(HuMAbs)，这些抗体可以中和LASV的传染性，并保护豚鼠免受LASV引起的致命疾病的影响。Zalgen的独家技术用于快速(&lt；3个月)开发表达稳定细胞系的高产量抗体，确保为一级生物防御市场和二级公共卫生市场生产治疗性抗体的成本降至最低。在里程碑1中提出的研究中，我们将在适应哺乳动物细胞瞬时表达平台的无血清介质中产生多毫克量的IgG1和IgG4 LASV huMAb亚型，用于纯化并进行体外测试。我们还将使用huMAb进行一系列表征分析，重点是识别具有广泛LASV中和潜力和低IC50的huMAb或huMAb组合。其他研究将建立与相关的新世界和旧世界阿拉伯病毒蛋白的交叉反应谱，以及LASV huMAb与人细胞膜磷脂的反应性。在里程碑2中，我们将在化学定义的无血清培养基中建立CHO或NS0稳定的细胞系，并选择高产菌株。有限稀释细胞克隆(LDCC)将用于分离稳定的生产级CD-SFM-CHO或CD-SFM-NS0克隆。我们还计划为感兴趣的克隆细胞系建立加入细胞库(ACB)，并转化为QA/QC生产，以产生克量的候选HuMAb，用于体内研究。在里程碑3中，我们将评估LASV huMAb亚型在豚鼠(GP)中的药代动力学。我们还将通过最佳给药途径，在全科医生中使用两个主要候选LASV huMAb或huMAb组合进行挑战-治疗剂量发现研究。接下来，我们将通过最佳确定的给药途径，在整个致死性感染时间线的GP研究中确定LASV huMAb或huMAb组合在良好实验室操作规范(GLP)GP研究中的治疗效果。GP研究的结果将指导在GLP非人灵长类(NHP)研究中评估两种单一LASV huMAb(IgG1或IgG4)或huMAb组合在单一静脉、腹膜或皮下给药后的药代动力学。我们还将在GLP NHP研究中进行LASV huMAb或huMAb组合的治疗效果研究，在感染后的不同时间，通过最佳给药途径进行两次剂量的致命感染时间线研究。这项拟议的努力将导致一类新的 LF的免疫治疗。