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: 8793681
• 负责人: Robert F Garry
• 金额: $ 101万
• 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 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8793681
• 关键词: Achievement; Africa; Antibodies; Area; Arenavirus; B-Lymphocyte Epitopes; Biological Assay; Biological Warfare; Biotechnology; Blocking Antibodies; Categories; Cavia; Cell Line; Cell membrane; Clinical Research; Clinical Trials; Clone Cells; Developed Countries; Development; Diagnostic; Disease; Dose; Drug Kinetics; Ensure; Evaluation; Future; Generations; Goals; Human; IgG1; IgG4; Immunologist; Immunotherapeutic agent; In Vitro; Individual; Infection; Inhibitory Concentration 50; Intravenous; Laboratories; Laboratory Research; Lassa Fever; Lassa virus; Lead; Life; Mammalian Cell; Marketing; Medical; Molecular Biology Techniques; Monoclonal Antibodies; Mus; National Institute of Allergy and Infectious Disease; Nigeria; Pharmaceutical Preparations; Phospholipids; Production; Productivity; Proteins; Public Health; Recombinant Proteins; Resources; Route; Scientist; Serum-Free Culture Media; Sierra Leone; Technology; Texas; Therapeutic; Therapeutic antibodies; Time; TimeLine; Toxicology; Treatment Efficacy; Universities; Virus Diseases; base; biodefense; biothreat; cell bank; combinatorial; cost; cross reactivity; good laboratory practice; human monoclonal antibodies; in vitro testing; in vivo; interest; intraperitoneal; large scale production; milligram; nonhuman primate; novel vaccines; point of care; pre-clinical; 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）：本项目将开发用于生物防御拉沙病毒（LASV）的免疫治疗药物，LASV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类制剂。LF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。该提案的目标是在治疗LF的一类新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得克萨斯大学医学分支的病毒学家和免疫学家的专业知识，以及Zalgen的科学家，Zalgen是一家专门开发免疫治疗，快速诊断和新疫苗平台的生物技术公司，用于全球影响的新兴病毒性疾病。这里提出的研究将利用我们最近的进展，包括鉴定主要候选人的人单克隆抗体（huMAbs），中和LASV感染性和保护豚鼠免受LASV诱导的致命疾病。Zalgen的独家技术用于快速开发（<3个月）高产抗体表达稳定细胞系，确保生产治疗性抗体的成本最小化，适用于初级生物防御市场和二级公共卫生市场。在MILESTONE 1中拟定的研究中，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多毫克量的IgG 1和IgG 4 LASV huMAb亚型，用于纯化以进行体外检测。我们还将使用huMAb进行一组表征试验，重点是鉴定具有广泛LASV中和潜力和低IC 50的huMAb或huMAb组合。其他研究将建立与相关Ne和旧世界沙粒病毒蛋白的交叉反应性特征以及LASV huMAb与人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NS 0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-NS 0克隆。我们还计划为感兴趣的克隆细胞系生成入组细胞库（ACB），并转换为QA/QC生产，以生成克量的候选huMAb用于体内研究。在MILESTONE 3中，我们将在豚鼠（GP）中评价LASV huMAb亚型的药代动力学。我们还将使用两种主要候选LASV huMAb或huMAb组合在GP中进行激发-治疗剂量探索研究，随后通过最佳递送途径进行单次给药。接下来，我们将在药物非临床研究质量管理规范（GLP）GP研究中，通过最佳确定的给药途径，在致死性感染时间轴的整个过程中确定LASV huMAb或huMAb组合的治疗功效。GP研究的结果将指导GLP非人灵长类动物（NHP）研究中两种单LASV huMAb（IgG 1或IgG 4）或huMAb复方制剂单次静脉、腹膜内或皮下给药后的药代动力学评价。我们还将在GLP NHP研究中，在感染后不同时间，通过最佳递送途径给药两次后，在整个致死性感染时间轴过程中，用LASV huMAb或huMAb组合进行治疗功效研究。这一提议的努力将导致一类新的 LF的免疫治疗。