Development of Protein Like Polymer Therapeutics for Modulating the Nrf2/Keap1 Protein Protein Interaction in Neurodegenerative Diseases

项目总结 人们对开发针对核间蛋白质-蛋白质相互作用的治疗方法非常感兴趣 神经退行性变的因子(红系2)样2(Nrf2)和海带样ECH相关蛋白1(Keap1) 疾病。成功和选择性地抑制Keap1/Nrf2结合的治疗方法将增强细胞 抗氧化反应产生神经保护作用。这种对Keap1/Nrf2相互作用的抑制可能 彻底改变我们治疗阿尔茨海默病等多种神经退行性疾病的能力。抑制性 多肽疗法越来越受到人们的关注，但它们的药代动力学曲线有限，细胞 渗透性和有效性。所提出的方法利用高密度刷子克服了这些挑战 能够与Keap1结合并取代细胞保护性转录因子的聚合物结构， NRF2.这些独特的结构被称为类蛋白质聚合物(PLP)，显示出特定的Nrf2衍生多肽 疏水合成聚合物核心周围的序列。与仅有多肽相比，PLP展示了 抵抗蛋白质分解，改善药代动力学，生物活性和有效的细胞摄取。PLP平台 是为了开发两种新型的Keap1抑制剂，以克服目前的挑战 限制Keap1/Nrf2靶向治疗。Keap1-目标PLP是这里的重点，因为文档中 Nrf2的神经保护作用和神经退行性疾病中大量未得到满足的临床需求。 一个关键的重点将是开发PLP，以竞争性地抑制Keap1并随后建立 它们作为Nrf2激活剂的有效性。通过计算机模拟鉴定的多肽将被结合到PLP中，然后 通过体外和体内分析来确定生物活性和药代动力学特性。PLP可以是 用与蛋白质相似的形态和分子量制备的。因此，我们将探讨 不同的分子量，从而聚合物长度，对Keap1抑制和生物活性。此外，PLP还可以 用多个基于多肽的结构域制备。这种多功能与多价性相结合将被使用 开发能够跨越并选择性结合两个Keap1结合位点的Nrf2模拟物。最后，PLP 平台将被用来设计一类能够异双功能结合的Keap1-抑制剂，使能 靶向降解Keap1。这些PLP被设计为针对嵌合体的自噬(AUTAC)，将 选择性地以Keap1为靶点，并通过自噬途径运送蛋白质进行降解。 拟议的研究计划旨在解决目前翻译治疗靶向面临的差距。 Keap1/Nrf2相互作用。拟议的研究将在内森博士的指导下进行 Gianneschi和Jeffrey Johnson博士在材料科学/化学和 神经生物学，并有权获得西北大学和 威斯康星大学。拟议的培训计划将把申请者培养成一名独立的研究人员 并培养临床技能，为成为一名内科科学家开发新的治疗平台做准备。

PROJECT SUMMARY There is significant interest in developing therapeutics targeting the protein-protein interaction between nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and kelch-like ECH-associating protein 1 (Keap1) for neurodegenerative disease. A therapeutic that successfully and selectively inhibits Keap1/Nrf2 binding would enhance the cellular antioxidant response leading to a neuroprotective effect. Such inhibition of the Keap1/Nrf2 interaction could revolutionize our ability to treat multiple neurodegenerative diseases such as Alzheimer’s Disease. Inhibitory peptide therapeutics are of increasing interest but are limited in their pharmacokinetic profiles, cellular penetration, and efficacy. The proposed approach overcomes these challenges utilizing high-density brush polymer structures that are capable of engaging Keap1 and displacing the cellular protective transcription factor, Nrf2. These unique structures, termed protein-like polymers (PLPs), display specific Nrf2-derived peptide sequences around a hydrophobic synthetic polymer core. Compared to peptides alone, PLPs demonstrate resistance to proteolysis, improved pharmacokinetics, bioactivity and efficient cellular uptake. The PLP platform is proposed here for the development of two novel types of Keap1-inhibitors that overcome the current challenges limiting Keap1/Nrf2 targeted therapeutics. Keap1-targeting PLPs are the focus here due to the documented neuroprotective role of Nrf2 and the large unmet clinical need in neurodegenerative disease broadly. A key focus will be on the development of PLPs to competitively inhibit Keap1 and subsequently establish their efficacy as Nrf2 activators. Peptides identified via in silico modeling will be incorporated into PLPs and then evaluated by in vitro and in vivo analysis to determine bioactivity and pharmacokinetic properties. PLPs can be prepared with morphologies and molecular weights similar to proteins. Therefore, we will explore the effects of varying molecular weight, and thus polymer length, on Keap1-inhibition and bioactivity. In addition, PLPs can be prepared with multiple peptide-based domains. This multifunctionality combined with multivalency will be used to develop Nrf2 mimetics capable of spanning and selectively binding both Keap1 binding sites. Finally, the PLP platform will be used to design a class of Keap1-inhibitors capable of heterobifunctional binding, enabling targeted degradation of Keap1. These PLPs, designed as autophagy targeting chimeras (AUTACs), will selectively target Keap1 and shuttle the protein for degradation through the autophagy pathway. The proposed research plan seeks to address the current gaps facing translational therapeutics targeting the Keap1/Nrf2 interaction. The proposed research will be conducted under the guidance of Dr. Nathan Gianneschi and Dr. Jeffrey Johnson to establish technical skills in both materials science/chemistry and neurobiology respectively and with access to expertise and equipment at Northwestern University and the University of Wisconsin. The proposed training plan will develop the applicant into an independent researcher and foster clinical skills in preparation for a career as a physician scientist developing novel therapeutic platforms.