Brain-cell penetrating antibodies for treatment of progressive multiple sclerosis

抽象的。中枢神经系统（CNS）的物理损伤，包括细菌或病毒感染，可诱导 慢性神经炎症，被认为持续一个人的一生。的其他 在炎症事件中，人们认识到补体途径的急性和慢性激活都起着重要的作用。 通过诱导神经元细胞损失和突触修剪在继发性脑损伤的发展中起作用。 补体过度激活也与不可逆进展的病理学密切相关 多发性硬化症（MS）是一种常见的中枢神经系统炎症和神经退行性疾病。我们假设 补体系统的治疗性抑制和神经生长的同时刺激可以防止 中枢神经系统组织损伤，减缓甚至阻断MS的进展。 目前，将药物递送至CNS的主要障碍是存在选择性可渗透的血脑 血脑屏障（BBB），限制血液传播的蛋白质进入中枢神经系统。为了解决这个问题，我们最近 开发了几种有效的骆驼衍生纳米抗体。第一组纳米抗体可以抑制补体 第二组包括酪氨酸激酶受体TrkB激动剂，其模拟酪氨酸激酶受体TrkB的作用。 脑源性神经营养因子（BDNF），一种促进神经元存活的脑生长因子， 可塑性和神经发生。在这里，我们建议进一步工程化这些纳米抗体，以促进它们的交叉 从而获得更有效地抑制补体级联和/或刺激神经的能力 在CNS内生长。 在I期期间，将产生双特异性纳米抗体并在体外细胞功能测定中验证。 治疗功效将在良好表征的进行性MS的鼠模型中进一步验证， Theiler小鼠脑脊髓炎病毒诱导的脱髓鞘疾病（TMEV-IDD）。为此，小鼠将 用对照纳米抗体、补体抑制剂纳米抗体、TrkB激动性纳米抗体或其组合治疗 后两者中的一个。一旦在TMEV-IDD中确认治疗功效，将施用骆驼科动物衍生的纳米抗体。 人源化以降低人中的抗原性。 监测给药小鼠的统计学显著改善对残疾进展和CNS的影响 病理学将是第二阶段提交的基础。第二阶段的目标是1）重新验证治疗方法 人源化抗体在更广泛的实验中的功效，包括对 性别、年龄、剂量范围和延迟治疗，即，感染后30天后，残疾进展， 疾病病理学和恢复; 2）根据当前良好生产建立生产方案 实践条件和; 3）定义多个研究中的生物学反应、PK/PD、剂量范围和毒理学 模型动物，包括非人类灵长类动物的毒理学研究。

Abstract. Central Nervous System (CNS) physical injuries, including bacterial or viral infection, can induce chronic neuroinflammation that is believed to persist for the lifetime of an individual. Among the other inflammatory events, it is recognized that both acute and chronic activation of the complement pathway plays a role in the development of secondary brain injuries by inducing neuronal cell loss and synaptic pruning. Complement over-activation is also firmly implicated in the pathology that underlies the irreversible progression of multiple sclerosis (MS), a common inflammatory and neurodegenerative disease of the CNS. We hypothesize that therapeutic inhibition of the complement system and concurrent stimulation of nerve growth may prevent CNS tissue damage and slow or even block the progression of MS. Currently, the main obstacle for drug delivery to the CNS is the presence of a selectively permeable blood-brain barrier (BBB), limiting blood-borne proteins' entryinto the CNS. To overcome this issue, we have recently developed several potent camelid-derived nanobodies. The first group of nanobodies can inhibit complement activation, whereas the second group comprises tyrosine kinase receptor TrkB agonists that mimic the action of brain-derived neurotrophic factor (BDNF), a growth factor in the brain that promotes neuronal survival, synaptic plasticity and neurogenesis. Here, we propose to engineer these nanobodies further to facilitate their crossing of the BBB, thereby gaining the ability to more effectively inhibit the complement cascade and/or stimulate nerve growth within the CNS. During phase I, bispecific nanobodies will be produced and validated in in vitro cellular functional assays. Therapeutic efficacy will be further validated in a well-characterized murine model of progressive MS, the Theiler's murine encephalomyelitis virus-induced demyelinating disease (TMEV-IDD). To this end, mice will be treated with control nanobodies, a complement inhibitor nanobody, TrkB agonistic nanobody, or a combination of the latter two. Once therapeutic efficacy is confirmed in TMEV-IDD, the camelid-derived nanobodies will be humanized to reduce antigenicity in humans. Statistically significant improvement in treated mice monitored as an impact on disability progression and CNS pathology will be the foundation for a phase II submission. The goals of phase II are 1) revalidate the therapeutic efficacy of humanized antibodies in more extensive experiments, including a detailed analysis of the effect of sex, age, dose-ranges and delayed treatments, i.e., later than 30 days post-infection, on disability progression, disease pathology and recovery; 2) to establish manufacturing protocols under current Good Manufacturing Practice conditions and; 3) to define the biological response, PK/PD, dose-ranging and toxicology in multiple model animals, including toxicology studies in non-human primates.