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

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 目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，因为这些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，可以成功治疗中风。随着提议的 我们寻求开发有效的、选择性的肽酶神经溶素 (Nln) 小分子激活剂， 将用作研究工具和主导化学实体，以推动药物发现过程的发展 新型药物。我们实验室最近发表的初步研究已确定 Nln 是大脑的自我调节机制之一。 保护机制，有助于中风后大脑的保存和恢复。功能意义 Nln 在中风后大脑中的作用是基于其灭活多种神经毒性物质并产生三种脑毒性物质的能力。 保护性/再生性神经肽，从大量实验和临床研究中已知， 有助于中风的结果。基于这一证据，我们将 Nln 视为参与大脑恢复的中枢肽酶 中风后的机制。在此合作应用中，我们将利用我们在药物多个方面的专业知识 发现过程并将开发有效的“类药物”小分子，可以选择性地提高催化效率 Nln 可用作中风后脑保护和恢复的实验治疗剂。该提案有 根据我们令人信服的实验数据制定，表明 Nln 的催化活性可以通过两个增强 结构相关的二肽和独特的非肽化学型。我们的研究表明了拟议研究的可行性 对已鉴定的 Nln 激活剂进行了初步结构-活性关系研究，并通过两种不同的体内实验 小鼠中风模型表明，中风后抑制内源性 Nln 会加重中风损伤，而 Nln 在大脑中的过度表达或将其输送到中风后大脑可显着改善中风结果。目标 该提案将通过三个综合目标来实现：（1）设计和表征多样化和专有的 基于三个活性命中和引导生物测定的化合物库，以识别关键的功能残基相互作用 Nln 结合位点内，并开发具有“药物样”功能的高效、脑渗透性、选择性 Nln 激活剂 特性; (2) 进行生化和结构研究，以表征所鉴定的 Nln 的激活机制 激活器利用； (3) 确定 Nln 激活剂在中风后脑保护和恢复中的治疗潜力 小鼠中风模型。这项工作具有高度创新性，因为科学文献中没有描述 Nln 的激活剂。 文献和此类化合物从未被认为具有治疗潜力。协作调查组， 由药物化学和药物发现、晶体学和结构生物学、酶生物化学领域的专家组成 和药理学、血脑屏障生理学和中风药理学，非常有资格进行拟议的 研究。我们的长期目标是将领先的 Nln 激活剂从实验室转移到临床并开发有效的疗法， 这将改变大量中风患者目前的治疗方式。

项目成果

Small molecule neurolysin activators, potential multi-mechanism agents for ischemic stroke therapy.

小分子溶神经素激活剂，缺血性中风治疗的潜在多机制药物。

• DOI: 10.1080/14728222.2022.2077190
• 发表时间: 2022
• 期刊: Expert opinion on therapeutic targets
• 影响因子: 5.8
• 作者: Esfahani,ShivaHadi;Abbruscato,ThomasJ;Trippier,PaulC;Karamyan,VardanT
• 通讯作者: Karamyan,VardanT

The role of peptidase neurolysin in neuroprotection and neural repair after stroke.

• DOI: 10.4103/1673-5374.284904
• 发表时间: 2021-01
• 期刊: Neural regeneration research
• 影响因子: 6.1
• 作者: Karamyan VT

The Effect of Histone Deacetylase Inhibitors Panobinostat or Entinostat on Motor Recovery in Mice After Ischemic Stroke.

• DOI: 10.1007/s12017-021-08647-1
• 发表时间: 2021-12
• 期刊: Neuromolecular medicine
• 影响因子: 3.5
• 作者: Al Shoyaib A;Alamri FF;Syeara N;Jayaraman S;Karamyan ST;Arumugam TV;Karamyan VT
• 通讯作者: Karamyan VT

Discovery of the Next Generation of Non-peptidomimetic Neurolysin Activators with High Blood-Brain Barrier Permeability: a Pharmacokinetics Study in Healthy and Stroke Animals.

具有高血脑屏障渗透性的下一代非拟肽神经溶素激活剂的发现：健康和中风动物的药代动力学研究。

• DOI: 10.1007/s11095-023-03619-5
• 发表时间: 2023
• 期刊: Pharmaceutical research
• 影响因子: 3.7
• 作者: Zhang,Yong;Sharma,Sejal;Jonnalagadda,Shirisha;Kumari,Shikha;Queen,Aarfa;Esfahani,ShivaHadi;Archie,SabrinaRahman;Nozohouri,Saeideh;Patel,Dhavalkumar;Trippier,PaulC;Karamyan,VardanT;Abbruscato,ThomasJ
• 通讯作者: Abbruscato,ThomasJ

Delayed atomoxetine or fluoxetine treatment coupled with limited voluntary running promotes motor recovery in mice after ischemic stroke.

• DOI: 10.4103/1673-5374.301031
• 发表时间: 2021-07
• 期刊: Neural regeneration research
• 影响因子: 6.1
• 作者: Alamri FF;Al Shoyaib A;Syeara N;Paul A;Jayaraman S;Karamyan ST;Arumugam TV;Karamyan VT
• 通讯作者: Karamyan VT