Discovery of therapeutic nanobodies targeting brain TNF-α for the treatment of Alzheimer Disease

项目摘要 散发性晚发性阿尔茨海默病(LOAD)是最常见的衰老依赖形式 神经退行性疾病。负荷与细胞外淀粉样蛋白沉积有关 Aβ多肽和磷酸盐形成的斑块和神经元内神经原纤维缠结(NFT)。 Tau，分别在中枢神经系统(CNS)。尽管有强烈的共识认为 有毒形式的β和tau会导致认知障碍，即停止或逆转认知的努力 以Aβ、斑块和缠结为靶点的负荷患者下降无效， 强调未得到满足的、不是基于负荷相关的治疗方法的需求 解剖病理学。 一些数据涉及神经炎症、小胶质细胞功能障碍和超生理学肿瘤坏死因子α 活动在LOAD发病机制中的作用。其中包括达达米奥博士产生的新证据， NanoNewron LLC的联合创始人，表明超生理学肿瘤坏死因子-α增加 兴奋性/抑制性平衡和损害突触的一种形式的长时程增强(LTP) 代表学习和记忆的细胞模型的可塑性。这些变化发生得较早。 并且不受Aβ水平和脑病理变化的影响。重要的是，重置肿瘤坏死因子α 低剂量抗大鼠肿瘤坏死因子-α中和抗体在生理水平上的活性 使这些更改正常化。 有证据表明，FDA批准的生物肿瘤坏死因子α脊髓周围和鞘内给药- 特异性抑制物(TNFi)改善LOAD患者的认知进一步强调 负载靶向超生理学肿瘤坏死因子α的治疗潜力。不幸的是，FDA- 批准的生物肿瘤坏死因子具有有限的血脑屏障(BBB)渗透，因为 尺码。 为了克服这一血脑屏障穿透障碍，NanoNewron LLC提出了一种第一阶段STTR 表征中和肿瘤坏死因子-α活性的抗肿瘤坏死因子-α驼胶体纳米小体的项目(TnFi- NABS)，目的是预防、阻止或扭转负荷。考虑到它的小尺寸和 与传统抗体相比，纳米抗体可以更快地穿透中枢神经系统。 此外，通过将纳米体靶向转铁蛋白，可以增加血脑屏障的通透性 微血管内皮细胞表面的受体，有利于跨细胞作用，和/或通过 增加等电点。因此，NanoNewron还开发了单价TfR-NaB 这将被用来有效地将TNFi-Nabs运送到CNS。这项研究的结果将 作为NanoNewron努力开发TNFi-NABS疾病修饰剂的垫脚石 治疗候选药物。

Project Summary Sporadic, late onset Alzheimer disease (LOAD) is the most common form of aging-dependent neurodegenerative disorder. LOAD is associated with deposition of extracellular amyloid plaques and intra-neuronal neurofibrillary tangles (NFT), formed by Aβ peptides and phosphor- tau, respectively, in the central nervous system (CNS). Although there is strong consensus that toxic forms of Aβ and tau cause cognitive impairments, efforts to halt or reverse cognitive decline in LOAD patients targeting Aβ, plaques and tangles have been ineffective and emphasize an unmet need for therapeutic approaches that are not based on LOAD-associated anatomical pathologies. Several data implicate neuroinflammation, microglia dysfunction and supraphysiological TNFα activity in LOAD pathogenesis. These include new evidence generated by Dr. D’Adamio, the cofounder of NanoNewron LLC, indicating that supraphysiological TNF-α increases excitatory/inhibitory balance and impairs Long-Term Potentiation (LTP), a form of synaptic plasticity that represents a cellular model for learning and memory. These changes occur early and are independent of changes in Aβ levels and brain pathology. Importantly, resetting TNFα activity at physiological levels using low doses of anti-rat-TNF-α neutralizing antibody normalizes these alterations. The evidence that peri-spinal and intrathecal administration of FDA-approved biologic TNFα– specific inhibitors (TNFI) improved cognition in LOAD patients further emphasizes the therapeutic potential of targeting supraphysiological TNFα in LOAD. Unfortunately, FDA- approved biologic TNFIs have limited blood-brain barrier (BBB) penetration because of the large size. To overcome this BBB-penetrance obstacle, NanoNewron LLC proposes a Phase I STTR project to characterize anti-TNF-α camelid nanobodies that neutralize TNF-α activity (TNFI- Nabs), with the goal of preventing, arresting or reversing LOAD. Given the small size and structure, nanobodies can penetrate the CNS much more rapidly than conventional antibodies. Moreover, BBB permeability can be increased by targeting the nanobody to the transferrin receptors on the surface of microvascular endothelial cells, favoring transcytosis, and/or by increasing the isoelectric point. Thus, NanoNewron has also developed monovalent TfR-Nabs that will be used to shuttle TNFI-Nabs efficiently into the CNS. The results of this study will serve as a stepping-stone for NanoNewron’s efforts to develop TNFI-Nabs disease modifying therapeutic candidates for LOAD.