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

项目摘要 人们对开发靶向细胞核内蛋白质-蛋白质相互作用的治疗剂非常感兴趣。 因子（红细胞衍生2）样2（Nrf 2）和kelch样ECH相关蛋白1（Keap 1）用于神经退行性变 疾病成功地和选择性地抑制Keap 1/Nrf 2结合的治疗剂将增强细胞免疫应答。 抗氧化反应导致神经保护作用。这种抑制Keap 1/Nrf 2相互作用的作用可能 彻底改变我们治疗多种神经退行性疾病如阿尔茨海默病的能力。抑制 肽治疗剂受到越来越多的关注，但其药代动力学特征、细胞毒性、细胞毒性和生物相容性受到限制。 渗透和功效。所提出的方法克服了这些挑战，利用高密度刷 能够接合Keap 1并置换细胞保护性转录因子的聚合物结构， Nrf2。这些独特的结构，称为蛋白质样聚合物（PLPs），显示特定的Nrf 2衍生肽 疏水合成聚合物核周围的序列。与单独的肽相比，PLP证明了 抗蛋白水解、改善的药代动力学、生物活性和有效的细胞摄取。PLP平台 在这里提出了两种新型的Keap 1抑制剂，克服目前的挑战的发展 限制了Keap 1/Nrf 2靶向治疗。Keap 1-定位PLP是这里的重点，因为记录了 Nrf 2的神经保护作用以及神经退行性疾病中大量未满足的临床需求。 一个关键的重点将是开发PLPs，以竞争性抑制Keap 1，并随后建立 它们作为Nrf 2激活剂的功效。通过计算机模拟鉴定的肽将掺入PLP中，然后 通过体外和体内分析评价以确定生物活性和药代动力学性质。PLP可以是 其形态和分子量与蛋白质相似。因此，我们将探讨 改变分子量，从而改变聚合物长度，对Keap 1抑制和生物活性的影响。此外，PLP可以是 用多个基于肽的结构域制备。这种多功能性与多价性的结合将被用于 开发能够跨越并选择性结合两个Keap 1结合位点的Nrf 2模拟物。最后，PLP 该平台将用于设计一类能够异双功能结合的Keap 1抑制剂， 靶向降解Keap 1。这些PLPs被设计为自噬靶向嵌合体（AUTAC）， 选择性地靶向Keap 1，并通过自噬途径穿梭蛋白质进行降解。 拟议的研究计划旨在解决目前转化治疗靶向所面临的差距 Keap 1/Nrf 2相互作用。拟议的研究将在内森博士的指导下进行 Gianneschi和Jeffrey约翰逊博士在材料科学/化学和 神经生物学分别与访问的专业知识和设备在西北大学和 威斯康星州大学。拟议的培训计划将使申请人成为一名独立的研究人员 培养临床技能，为成为一名医生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.