Structure-Guided Design of Broadly Neutralizing Lassa Virus BiSpecific Antibodies

广泛中和拉沙病毒双特异性抗体的结构引导设计

• 批准号: 10306341
• 负责人: Luis Manuel Branco
• 金额: $ 119.3万
• 依托单位: ZALGEN LABS, LLC
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-24 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10306341
• 关键词: Achievement; Address; Africa; African; Animal Model; Animals; Antibodies; Antibody Therapy; Antibody titer measurement; Antiviral Agents; Architecture; Biological Products; Biotechnology; Bispecific Antibodies; Cavia; Chemistry; Clinical Research; Complex; Convalescence; Crystallization; Data; Derivation procedure; Development; Disease; Dose; Drug resistance; Economics; Engineering; Epitopes; Escape Mutant; Evaluation; GP2 gene; GTPBP1 gene; Geography; Glycoproteins; Government; Hematology; Human; Human Engineering; Immunocompromised Host; Immunotherapeutic agent; Individual; Infection; Infection prevention; Intellectual Property; Intoxication; Lassa Fever; Lassa virus; Macaca fascicularis; Metabolic; Modality; Modeling; Monoclonal Antibodies; National Institute of Allergy and Infectious Disease; Ownership; Patients; Persons; Pharmacology and Toxicology; Preparation; Readiness; Recommendation; Reporting; Resistance; Resolution; Resource-limited setting; Rodent; Structure; Symptoms; Technology; Testing; Therapeutic; Therapeutic antibodies; Time; Vaccines; Viral; Viral Antigens; Viral Hemorrhagic Fevers; Virus; base; commercialization; design; emerging pathogen; human monoclonal antibodies; improved; interest; lead candidate; monomer; mutant; neutralizing monoclonal antibodies; nonhuman primate; novel; pre-clinical; preclinical evaluation; prevent; programs; prophylactic; rational design; social; therapeutic candidate; weapons

“Structure-Guided Design of Broadly Neutralizing Lassa Virus BiSpecific Antibodies” ABSTRACT Lassa fever (LF) is an often-fatal viral hemorrhagic fever (VHF) that is endemic in West Africa where it causes significant social and economic disruption. The lack of an approved therapeutic or vaccine, recorded geographic expansion of rodent reservoirs, ease of procurement and weaponization of the virus, and the recent emergence of new Lassa virus (LASV) strains support recommendations for enhanced preparedness for LF. We isolated and characterized 113 human monoclonal antibodies (hMAbs), the first large panel of human antibodies against LASV described. We found that the most potent neutralizing hMAbs target quaternary epitopes that require both GP1 and GP2 subunits of each monomer in the glycoprotein complex (GPC) trimer. LASV is genetically diverse with four distinct lineages present in West Africa. Some hMAbs neutralized all 4 LASV lineages. Challenge of outbred guinea pigs (GP) in a model of lethal LF informed the down-selection of broadly neutralizing hMAbs (BNhMAbs) for studies in a nonhuman primate (NHP) model, Cynomolgus macaques. A combination of 3 BNhMAbs, each with broad neutralizing activity and recognition of distinct epitopes on the LASV GPC, rescued 100% of NHPs even after delay in the start of treatment to 8 days post-infection, a time when the animals displayed severe hematological and metabolic dysregulation. The 3 BNhMAb cocktail conferred 100% protection in NHP against lethal challenge with LASV strains from lineages II and IV. We now propose to utilize the structural information of BNhMAbs complexed with GPC to engineer human bi-specific antibodies (BsAbs) that span two highly protective epitopes, thereby reducing the number of molecules required to confer superior protection against LF. Preliminary results in LASV- challenged GP suggest that targeting quaternary neutralizing epitopes in the base of GPC with a bi-specific antibody results in superior protection, even at 10-fold lower doses than previoulsy tested for individual BNhMAbs (Preliminary Results). Our proposed project meets the strict requirements of RFA-AI-17-026 in that the LASV BsAbs are based on previously identified, well-characterized, candidate therapeutic hMAbs against an NIAID listed emerging pathogen, LASV. The project will address a particular interest of RFA-AI- 17-026 for immunotherapeutics that would “enable prevention of infection or intoxication in the face of an immediate threat, protection of immunocompromised individuals, or post-exposure treatment to suppress infection and disease.” In Milestone 1 we will down-select BsAbs targeting base, middle, and cap neutralizing epitopes on LASV GPC. In Milestone 2 dose and dosing interval studies with mono and combination BsAb therapy in GP and NHP will be evaluated. In Milestone 3 Chemistry, Manufacturing and Control Data (CMC) will be established for leading BsAbs. In Milestone 4 we will perform Preclinical Pharmacology and Toxicology of BsAbs. At the conclusion of the proposed program we will complete preclinical evaluation of a first-in-class immunotherapeutic BsAb for the prophylactic and post-exposure treatment of LF. This application contains proprietary/priviledged information that Zalgen Labs and its subcontractors request not be released to persons outside the Government, except for the purposes of review and evaluations.

“结构引导的广泛中和lassa病毒双特异性抗体的设计” 抽象的 Lassa Fever（LF）是一种经常致命的病毒出血热（VHF），它是西非的内在 造成严重的社会和经济破坏。缺乏批准的疗法或疫苗，记录 啮齿动物储层的地理扩展，易于采购和武器化病毒以及 新的LASSA病毒（LASV）菌株的最新出现支持建议的建议 对于LF。我们隔离并表征了113种人类单克隆抗体（HMAB），这是第一个大面板 对LASV的人类抗体描述。我们发现最潜在的中和HMAB目标 糖蛋白络合物中每个月都需要GP1和GP2亚基的第四纪表位 （GPC）三聚体。 LASV在遗传上是多样的，西非有四个不同的谱系。一些HMAB 中和所有4个LASV谱系。在致命的LF模型中，差异豚鼠（GP）的挑战告诉了 在非人类灵长类动物（NHP）模型中进行广泛中和的HMAB（BNHMAB）的下和概述， cynomolgus猕猴。 3个BNHMAB的组合，每个BNHMAB具有广泛的中和活性和识别 LASV GPC上不同的表位，即使在治疗开始时延迟了100％的NHP 感染后的天数，动物表现出严重的血液学和代谢失调的时代。 3种BNHMAB鸡尾酒在NHP中授予100％的保护，以防止致命挑战，从 血统II和IV。现在，我们建议利用与GPC复合的BNHMAB的结构信息 工程师人类双特异性抗体（BSAB），跨越了两个高度保护的表位，从而减少了 会议对LF进行优越保护所需的分子数量。 lasv-的初步结果 受到挑战的GP表明，针对GPC底部的第四纪中和表位的靶向用双特异性 抗体可产生优越的保护，即使剂量低10倍，比为个体测试 BNHMAB（初步结果）。我们提议的项目符合RFA-AI-17-026的严格要求 LASV BSAB基于先前确定的，良好的候选疗法HMAB 针对NIAID列出的新兴病原体LASV。该项目将解决RFA-AI-的特别兴趣 17-026用于免疫治疗药，可以预防面对感染或中毒 立即威胁，保护免疫功能低下的个体或暴露后治疗以抑制 在里程碑1中，我们将靶向基础，中和帽子中和的选择BSABS LASV GPC上的表位。在Milestone 2剂量和剂量间隔研究中，单声道和BSAB组合 将评估GP和NHP的治疗。在里程碑3化学中，制造和控制数据（CMC） 将为领先的BSABS建立。在里程碑4中，我们将执行临床前药理学和 BSABS的毒理学。在拟议计划的结论结束时，我们将完成对A的临床前评估 LF的预防和暴露后治疗的第一类免疫治疗BSAB。 该应用程序包含Zalgen Labs及其分包商请求的专有/特权信息 除了审查和评估目的外，不要向政府以外的人释放。