Preclinical Evaluation of Advanced Pan-Lassa Immunotherapeutic Cocktails

拉沙热（LF）是西非流行的一种致命性病毒性出血热（VHF）。后一 来自西非患者的广泛中和人单克隆抗体（BNhMAb）， 存活的LF（Arevirumab-3）挽救了100%的非人灵长类动物（NHP），即使在延迟启动治疗后也是如此。 治疗至感染后8天。最近，我们解决了拉萨星融合前的晶体结构 病毒糖蛋白复合物（GPC）结合到BNhMAb，第一个结构的病毒粒子配置的 任何沙粒病毒的凝胶渗透色谱分析。GPC：BNhMAb复合物的另外的新解决的结构和GPC的衍生物 逃逸突变体为最佳配制的BNhMAb鸡尾酒的设计和评价提供了信息， 独立的中和表位。我们计划扩大这项广泛的工作，以支持完成 的临床前评价的Arevirumab-3和功能增强的衍生物与改进的 这类新型LF治疗剂的首次人体临床研究的药理学特性。在 根据具体目标1拟定的研究，我们将完成Arevirumab-3的临床前评价。 完成化学、生产和控制数据（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，将使提交一份 研究性新药（IND）申请用于Arevirumab-3的临床研究 LF动物模型中的非中和LASV huMAb将在特定目的中拟定的研究中进行评价 2.将在豚鼠中评价非中和LASV huMAb的保护效力并向下选择 猪如果鉴定出非中和保护性huMAb（NNMAb），则这类抗体的活性 将在LF的NHP模型中进一步评估，无论是单独还是合理设计的鸡尾酒。研究中 根据具体目标3的建议，我们将设计通过Fc效应器增强的免疫治疗剂， 计算优化、双特异性抗体工程（BsAbs）、“设计者”生产细胞系， 附加特定的聚糖和监测数据输入。用Fc增强免疫调节剂混合物 工程，BsAbs的评估，机器学习算法和基于CRISPR/Cas9的NS 0生成 或经设计者修饰的CHO生产细胞系将定义下一代LF的特征 免疫治疗对流行的和新出现的临床LASV毒株的监测将指导设计 LF免疫治疗剂具有稳健的临床特征和持续的效力。根据具体目标4，我们 提出研究以评估优化的BNhMAb、NNPhMAb和BsAb的体内保护和PK。体内 在已建立的LF模型中评价优化的LASV抗体将教育先进的临床前 具有改进的药理学性质的第二代免疫治疗剂的评价。研究 预计项目1中提出的方案将导致在第3年提交Arevirumab-3的IND申请， a在5年性能期结束时，第二代治疗药物的IND前。

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.