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沙病毒（LASV）的生物防御免疫疗法，拉沙病毒是一种沙粒病毒，是拉沙热（LF）的病原体。 LASV 是生物安全 4 级和 NIAID 生物防御 A 类药剂。 LF 是一个重大的公共卫生问题，导致整个西非广泛的生命损失和社会混乱。该提案的目标是在治疗 LF 的新型疗法的表征和临床前开发中执行关键步骤。这项工作结合了杜兰大学和德克萨斯大学医学分部的病毒学家和免疫学家的专业知识，以及 Zalgen 的科学家的专业知识。Zalgen 是一家生物技术公司，专门开发针对具有全球影响的新兴病毒性疾病的免疫疗法、快速诊断和新疫苗平台。这里提出的研究将利用我们的最新进展，包括鉴定主要候选人单克隆抗体 (huMAb)，这些抗体可以中和 LASV 感染性并保护豚鼠免受 LASV 诱发的致命疾病。 Zalgen 的独家技术可快速开发（<3 个月）高产抗体表达稳定细胞系，确保将初级生物防御市场和二级公共卫生市场的治疗性抗体生产成本降至最低。在 MILESTONE 1 提出的研究中，我们将在无血清培养基适应的哺乳动物细胞瞬时表达平台中产生数毫克量的 IgG1 和 IgG4 LASV huMAb 亚型，进行纯化以进行体外测试。我们还将对 huMAb 进行一组表征测定，重点是鉴定具有广泛 LASV 中和潜力和低 IC50 的 huMAb 或 huMAb 组合。其他研究将建立与相关新世界和旧世界沙粒病毒蛋白的交叉反应性特征以及 LASV huMAb 与人类细胞膜磷脂的反应性。在 MILESTONE 2 中，我们将在化学成分确定的无血清培养基中生成 CHO 或 NS0 稳定细胞系，并选择高产分离株。将进行有限稀释细胞克隆 (LDCC) 以分离稳定的生产级 CD-SFM-CHO 或 CD-SFM-NS0 克隆。我们还计划为感兴趣的克隆细胞系生成登记细胞库 (ACB)，并通过 QA/QC 进行生产，以生成用于体内研究的克级候选 huMAb。在 MILESTONE 3 中，我们将在豚鼠 (GP) 中评估 LASV huMAb 亚型的药代动力学。我们还将在全科医生中使用两种主要候选 LASV huMAb 或 huMAb 组合进行挑战-治疗剂量寻找研究，通过最佳递送途径进行单剂量给药。接下来，我们将通过最佳确定的给药途径，在良好实验室规范 (GLP) GP 研究中确定 LASV huMAb 或 huMAb 组合在整个致命感染时间线过程中的治疗效果。 GP 研究的结果将指导在 GLP 非人灵长类动物 (NHP) 研究中单次静脉内、腹膜内或皮下剂量后评估两种单一 LASV huMAb（IgG1 或 IgG4）或 huMAb 组合的药代动力学。我们还将在 GLP NHP 研究中，在整个致死性感染时间线的整个过程中，在感染后的不同时间，通过最佳给药途径注射两次剂量后，对 LASV huMAb 或 huMAb 组合进行治疗效果研究。这项拟议的努力将产生一类新的 LF 的免疫治疗。