Development and Characterization of Peptidomimetic Small Molecule Activators of Peptidase Neurolysin for Stroke Therapy

用于中风治疗的肽酶神经溶素的肽模拟小分子激活剂的开发和表征

• 批准号: 10753623
• 负责人: Thomas J Abbruscato
• 金额: $ 57.86万
• 依托单位: OAKLAND UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10753623
• 关键词: Development; Characterization; Peptidomimetic; Small; Molecule; Development; Characterization; Peptidomimetic; Small; Molecule

SUMMARY Current stroke research focuses more on understanding the brain’s self-protective and repair mechanisms. Detailed elucidation of these mechanisms is crucial as such knowledge could lead to development of therapeutic interventions which mimic or engage the brain’s self-protective/repair mechanisms and can lead to successful stroke therapy. With the proposed research we seek to develop potent and selective ‘drug-like’ small molecule activators of peptidase neurolysin (Nln) which will be used as research tools and lead chemical entities to move the drug discovery process forward for development of a novel class of drugs. Recently published and pilot studies from our laboratory have identified Nln as one of the brain’s self- protective mechanisms, functioning towards preservation and recovery of the brain after stroke. Functional significance of Nln in the post-stroke brain is based on its ability to inactivate several neurotoxic and generate three cerebro- protective/regenerative neuropeptides, which are known from numerous experimental and clinical studies to critically contribute to the outcome of stroke. Based on this evidence we view Nln as a central peptidase involved in brain restorative mechanisms following stroke. In this collaborative application we will leverage our expertise in multiple aspects of the drug discovery process and will develop potent ‘drug-like’ small molecules which can selectively enhance the catalytic efficiency of Nln and can be used as experimental therapeutic agents for post-stroke brain protection and recovery. This proposal has been formulated based on our compelling experimental data indicating that catalytic activity of Nln can be enhanced by two structurally related dipeptides and a distinct non-peptide chemotype. Feasibility of the proposed studies is shown by our initial structure-activity relationship studies of the identified Nln activators and by in vivo experiments in two different mouse stroke models indicating that inhibition of endogenous Nln after stroke aggravates stroke injury, whereas overexpression of Nln in the brain or its delivery to the post-stroke brain substantially improves stroke outcome. The goals of this proposal will be accomplished in three well-integrated aims: (1) design and characterize a diverse and proprietary library of compounds based on three active hits and guided bioassays to identify critical functional residue interactions within the Nln binding site, and to develop high-potency, brain-permeable, selective activators of Nln with ‘drug-like’ properties; (2) conduct biochemical and structural studies to characterize the activation mechanism that the identified Nln activators exploit; (3) determine the therapeutic potential of Nln activators in post-stroke brain protection and recovery using a mouse model of stroke. This work is highly innovative because there are no activators of Nln described in the scientific literature and such compounds were never considered to have therapeutic potential. The collaborative investigative team, comprising experts in medicinal chemistry and drug discovery, crystallography and structural biology, enzyme biochemistry and pharmacology, blood-brain barrier physiology and stroke pharmacology, is highly qualified to conduct the proposed studies. Our long-term goal is to translate the lead Nln activators from bench to bedside and develop an effective therapy, which would transform the current treatment modalities for a vast number of stroke patients.

概括 当前的中风研究更多地侧重于了解大脑的自我保护和修复机制。详细的 阐明这些机制至关重要，因为这种知识可能导致治疗干预措施的发展 模仿或参与大脑的自我保护/修复机制，并可能导致成功中风疗法。提议 我们寻求开发肽神经蛋白（NLN）的潜在和选择性的“药物样”小分子激活剂的研究 将用作研究工具，并领导化学实体推动药物发现过程的发展，以开发 新型药物。最近发表的和我们实验室的试点研究已将NLN确定为大​​脑的自我之一 保护机制，在中风后的保存和恢复方面发挥作用。功能意义 中风后大脑中的NLN是基于它使几种神经毒性失活并产生三个脑毒性的能力 保护性/再生性神经肽，从众多实验和临床研究中得知这些神经肽 有助于中风的结果。基于此证据，我们将NLN视为参与大脑修复的中央肽 中风后的机制。在此协作应用中，我们将利用我们在药物的多个方面的专业知识 发现过程，并将开发潜在的“药物状”小分子，这些分子可以选择性提高催化效率 NLN的摄入量，可用作触摸后脑保护和恢复的实验治疗剂。该提议有 我们是根据我们引人入胜的实验数据制定的，表明NLN的催化活性可以通过两个增强 与结构相关的二肽和独特的非肽化学型。我们提出的研究的可行性由我们的 已鉴定的NLN激活剂和通过两个不同的体内实验的初始结构活性研究研究 小鼠中风模型表明，中风后抑制内源性NLN会加剧中风损伤，而 NLN在大脑中的过表达或其向中风后大脑的传递可大大改善中风的结果。目标 该提案将以三个良好的目的来完成：（1）设计和表征多样化和专有的 基于三个主动命中和指导生物测定的化合物库，以识别关键的功能居住相互作用 在NLN结合位点内，并开发具有“药物样”的NLN的高功能，可渗透的，可渗透的，选择性的活化剂 特性; （2）进行生化和结构研究以表征鉴定出NLN的激活机制 激活剂利用； （3）确定NLN激活剂在冲程后脑保护和恢复中使用的治疗潜力 鼠标中风模型。这项工作具有很高的创新性，因为在科学中没有NLN的激活剂 文献和这种化合物从未被认为具有治疗潜力。协作调查团队， 包括医学化学和药物发现专家，晶体学和结构生物学，酶生物化学 和药理学，血脑屏障生理学和中风药理学，有资格进行拟议的 研究。我们的长期目标是将铅NLN激活剂从长凳转换为床边，并开发有效的疗法， 这将改变许多中风患者的当前治疗方式。