Preclinical Evaluation of Advanced Pan-Lassa Immunotherapeutic Cocktails

拉沙热是西非流行的一种常常致命的病毒性出血热。三者的组合 来自西非患者的广泛中和人类单抗(BNhMAbs) 存活的LF(Arevirumab-3)拯救了100%的非人类灵长类动物(HAP)，即使在延迟启动 至感染后8天开始治疗。最近，我们解决了LASSA的预聚体形式的晶体结构 病毒糖蛋白复合体(GPC)与BNhMAb结合，是病毒粒子构型的第一个结构 任何病毒的GPC。GPC的其他新解结构：BNhMAb络合物及其衍生物 逃逸突变体为最佳配方BNhMAb靶向鸡尾酒的设计和评估提供了信息 独立中和表位。我们计划对这一广泛的工作进行扩展，以支持完成 对Arevirumab-3及其功能增强的衍生物进行临床前评估 这类新的LF治疗药物的药理学特性是人类临床研究中的第一例。在……里面 在特定目标1下提出的研究，我们将完成Arevirumab-3的临床前评估。 完成化学、制造和控制数据(CMC)、临床前药理学和毒理学 LF的动物模型，包括预防间隔和复发评估，将使提交 新药(IND)在Arevirumab-3临床研究中的应用 肝功能衰竭动物模型中的非中和性LASV huMAb将在特定目的的研究中进行评估 2.对非中和LASV huMAb在豚鼠体内的保护效果进行评价和筛选 猪。如果鉴定出非中和保护性HuMAbs(NNPhuMAbs)，这类抗体的活性 将在LFNHP模型中进一步评估，无论是单独的还是在合理设计的鸡尾酒中。在研究中 在具体目标3下提出，我们将通过Fc效应器和 计算优化、双特异性抗体工程(BsAbs)、“设计者”生产细胞系 附加特定的糖链和监控数据输入。FC对免疫治疗鸡尾酒的强化作用 工程、BsAbs评估、机器学习算法以及基于CRISPR/Cas9的NS0生成 或者经过设计师修改的CHO生产单元线将定义下一代LF的特征 免疫疗法。对临床流行的和新出现的LASV毒株的监测将教育设计 具有强大的临床特征和持续效力的LF免疫疗法。在具体目标4下，我们 建议开展研究，评估优化的BNhMAbs、NNPhMAbs和BsAbs的体内保护和PK。活体内 在已建立的肝功能衰竭模型中评估优化的LASV抗体将培养高级临床前 药理特性改善的第二代免疫疗法的评价。这些研究 在项目1中提出的这些药物预计将导致在第3年提交Arevirumab-3的IND申请，以及 在5年的表演期结束时进行第二代治疗的预IND。

Lassa fever (LF) is an often-fatal viral hemorrhagic fever (VHF) endemic in West Africa. A combination of three broadly neutralizing human monoclonal antibodies (BNhMAbs) derived from West African patients who survived LF (Arevirumab-3) rescued 100% of nonhuman primates (NHPs), even after delay in initiation of treatment to 8 days post-infection. Recently, we solved the crystal structure of the pre-fusion form of the Lassa virus glycoprotein complex (GPC) bound to a BNhMAb, the first structure for the virion configuration of the GPC of any arenavirus. Additional newly solved structures for GPC:BNhMAb complexes and derivation of escape mutants have informed the design and evaluation of optimally formulated BNhMAb cocktails targeting independent neutralizing epitopes. We plan to expand upon this extensive body of work to support completion of pre-clinical evaluation of Arevirumab-3 and functionally enhanced derivatives with improved pharmacological properties toward first-in-human clinical studies with this novel class of LF therapeutics. In studies proposed under Specific Aim 1, we will complete the pre-clinical evaluation of Arevirumab-3. Completion of Chemistry, Manufacturing and Control Data (CMC), preclinical pharmacology and toxicology in animal models of LF, including prophylactic interval and recrudescence evaluations, will enable the filing of an Investigational New Drug (IND) application toward clinical studies with Arevirumab-3 The protective activity of non-neutralizing LASV huMAbs in animal models of LF will be evaluated in studies proposed in Specific Aim 2. The protective efficacy of non-neutralizing LASV huMAbs will be evaluated and down-selected in guinea pigs. If non-neutralizing protective huMAbs (NNPhuMAbs) are identified, the activity of this class of antibodies will be further evaluated in NHP models of LF, either individually or in rationally designed cocktails. In studies proposed under Specific Aim 3, we will design enhanced immunotherapeutics via Fc effector and computational optimization, bi-specific antibody engineering (BsAbs), “designer” production cell lines that attach specific glycans, and surveillance data inputs. Enhancement of immunotherapeutic cocktails with Fc engineering, evaluation of BsAbs, machine learning algorithms, and CRISPR/Cas9-based generation of NS0 or CHO production cell lines with designer modifications will define characteristics of next generation LF immunotherapeutics. Surveillance of circulating and emerging clinical LASV strains will educate the design of LF immunotherapeutics with robust clinical characteristics and sustained potency. Under Specific Aim 4, we propose studies to valuate in vivo protection and PK of optimized BNhMAbs, NNPhMAbs, and BsAbs. In vivo evaluation of optimized LASV antibodies in established models of LF will educate advanced pre-clinical evaluation of second generation immunotherapeutics with improved pharmacological properties. The studies proposed in Project 1 are expected to result in the filing of an IND application for Arevirumab-3 in Year 3 and a Pre-IND for a 2nd generation therapeutic at the conclusion of the 5-year performance period.