Primary sensory neuron-targeted block of Cav3.2 for treatment of chronic neuropathic pain

初级感觉神经元靶向的 Cav3.2 阻断治疗慢性神经性疼痛

• 批准号: 10452646
• 负责人: Quinn H Hogan
• 金额: $ 43.96万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452646
• 关键词: Absence of pain sensation; Acquired Immunodeficiency Syndrome; Affect; Afferent Neurons; Analgesics; Anatomy; Arthritis; Behavioral; Behavioral Assay; Biological; Cells; Chemotherapy-Oncologic Procedure; Chronic Disease; Data; Development; Diabetes Mellitus; Disease; Dose; Drug Kinetics; Electrophysiology (science); Event; Female; Gene Delivery; Generations; Genetic; Goals; Histologic; Hour; Human; In Vitro; Industry; Inflammation; Injections; Injury; Ion Channel Protein; Maintenance; Measures; Mediating; Mental Depression; Mental Health; Modeling; Molecular; Molecular Target; Nervous system structure; Neurons; Neuropathy; Nociception; Opioid; Overdose; Pain; Pain Measurement; Pain management; Peptide aptamers; Peptides; Peripheral; Peripheral Nervous System; Pharmacodynamics; Phase; Plasmids; Production; Protein Region; Proteins; Rattus; Recombinants; Risk; Role; Safety; Sample Size; Selection Criteria; Sensory; Signal Transduction; Silver Staining; Site; Spinal Ganglia; Stress; Structure; T-Type Calcium Channels; Testing; Therapeutic; Time; Tissues; Traumatic Nerve Injury; Tropism; Work; addiction; addiction liability; adeno-associated viral vector; age group; attenuation; base; behavior test; cell type; chronic neuropathic pain; chronic pain; chronic pain management; cost; design; experimental study; flexibility; in silico; in vivo; inhibitor; interest; male; nerve injury; neuronal excitability; novel; novel therapeutics; pain behavior; pain model; pain reduction; painful neuropathy; prediction algorithm; protein aminoacid sequence; protein protein interaction; response; sex; side effect; vector; voltage

Selective block of CaV3.2 in primary sensory neurons for treatment of chronic neuropathic pain Summary/Abstract: Chronic pain is a devastating problem, while opioid treatment of chronic pain has numerous risks, including misuse, overdose, and addiction, highlighting the need for new analgesic targets. The peripheral sensory nervous system (PSNS) is a particularly accessible site for devising new treatments, where the sensory neurons of the dorsal root ganglia (DRG) initiate nociception and have a central role in the development and maintenance of painful neuropathy. Sensory neuronal CaV3.2 T-type calcium channels regulate neuronal excitability and are a promising target for treatment of pain, but the development of selective CaV3.2 inhibitors has proved elusive. Small peptides, especially those derived from the natural proteins as functionally inhibitory peptide aptamers (iPAs), are recognized as being highly effective and selective, allowing blockade of specific pain molecular interactomes to reduce pain with minimal off-target effects. The analgesic iPA-sustained expression in the PSNS delivered by adeno-associated viral vectors (AAV) in the DRG is a safe and feasible path to chronic pain treatment with minimal side effects and no abuse/addiction liability. Molecular signaling interactions are often mediated by regions of proteins lacking a defined tertiary structure, known as protein intrinsically disordered regions (IDRs). Using established IDR prediction algorithms, we localized highly disordered regions in CaV3.2, and identified a novel 17mer peptide that produces sustained CaV3.2 T-type current inhibition and pain attenuation in a neuropathic pain rat model, demonstrating its therapeutic potential for pain treatment. The proposed Two-Phase study is to develop evidence that candidate CaV3.2iPAs delivered by AAV into DRG neurons have sufficient biological activity to justify further development as a novel analgesic approach. In Phase I (R66), a combined computational and experimental strategy will be used to design candidate CaV3.2iPAs and test their inhibition of T-type current by in vitro of cell-based studies. The milestone for advancement from R66 Phase to Phase II (R33) is identification of the top CaV3.2iPA leads that meet our selection criteria (higher potency and selectivity in block of CaV3.2) and of production of therapeutic AAV- CaV3.2iPAs. Phase II aims to apply AAV-CaV3.2iPAs in vivo by intraganglionic injection to evaluate critical features of AAV-CaV3.2iPAs in nerve injury-induced rat pain model, compared to controls The milestone for the end of Phase II is generation of sufficient data of in vivo efficacy and safety which will help us to make a go/no- go informed decision for further translational development. Our ultimate goal is to develop a novel therapeutics combined AAV-targeted gene delivery with potent and selective iPA block of CaV3.2 in anatomically segmental DRG to treat chronic neuropathic pain.

选择性阻断初级感觉神经元中的 CaV3.2 治疗慢性神经性疼痛 摘要/摘要：慢性疼痛是一个毁灭性的问题，而阿片类药物治疗慢性疼痛已经 多种风险，包括误用、过量和成瘾，凸显了对新镇痛目标的需要。 周围感觉神经系统（PSNS）是设计新疗法特别容易的部位， 背根神经节 (DRG) 的感觉神经元在此启动伤害感受，并在 疼痛性神经病的发展和维持。感觉神经元 CaV3.2 T 型钙通道 调节神经元兴奋性，是治疗疼痛的一个有希望的目标，但选择性的开发 CaV3.2 抑制剂已被证明难以捉摸。小肽，尤其是源自天然蛋白质的小肽 功能抑制肽适体 (iPA) 被认为是高度有效和选择性的，允许 阻断特定的疼痛分子相互作用组以减轻疼痛，同时将脱靶效应降至最低。镇痛药 由腺相关病毒载体 (AAV) 在 DRG 中传递的 PSNS 中 iPA 持续表达是安全的 以及副作用最小且无滥用/成瘾倾向的慢性疼痛治疗的可行途径。 分子 信号相互作用通常由缺乏明确三级结构的蛋白质区域介导，称为 蛋白质本质无序区域（IDR）。使用已建立的 IDR 预测算法，我们高度定位 CaV3.2 中的无序区域，并鉴定了一种新的 17mer 肽，可产生持续的 CaV3.2 T 型 在神经性疼痛大鼠模型中的电流抑制和疼痛减弱，证明了其治疗潜力 用于疼痛治疗。拟议的两阶段研究旨在开发候选 CaV3.2iPA 交付的证据 通过 AAV 进入 DRG 神经元具有足够的生物活性，可以证明作为新型镇痛药的进一步开发是合理的 方法。在第一阶段（R66）中，将使用计算和实验相结合的策略来设计 候选 CaV3.2iPA 并通过体外细胞研究测试其对 T 型电流的抑制作用。里程碑 从 R66 阶段到 II 期 (R33) 的进展是确定符合我们的要求的顶级 CaV3.2iPA 引线 选择标准（阻断CaV3.2的更高效力和选择性）和治疗性AAV-的生产 CaV3.2iPA。 II 期旨在通过节内注射在体内应用 AAV-CaV3.2iPA 来评估关键 与对照组相比，AAV-CaV3.2iPA 在神经损伤诱发的大鼠疼痛模型中的特征 第二阶段的结束是产生足够的体内功效和安全性数据，这将有助于我们做出是否可行的决定 做出明智的决定以进一步转化发展。我们的最终目标是开发一种新的疗法 将 AAV 靶向基因递送与解剖学节段中 CaV3.2 的有效且选择性 iPA 阻断相结合 DRG 治疗慢性神经性疼痛。