Selective actin remodeling of sensory neurons for acute pain management

项目摘要 迫切需要新的方法来治疗急性人类疼痛，而不会有物质使用的风险 疾病（SUD）。最有效的批准疼痛药物，包括麻醉剂和麻醉剂，是 不是神经元特异性的，因此遭受脱靶效应，如成瘾，运动神经元抑制， 破坏周围组织。当炎症发生时，肌动蛋白聚合发生在感觉细胞中， 神经元，导致嘌呤能受体的敏化和异常的疼痛行为。靶向肌动蛋白 重塑可能是减轻急性伤害性疼痛的有效方法，但没有小分子 对感觉神经元具有足够特异性的抑制剂，可正确调节细胞骨架。脉壳属 提出了一种新的治疗伤害性疼痛的方法，该方法基于一种创新的工程蛋白，称为N- 001，其选择性地靶向感觉神经元并且仅在细胞内水平起作用， 轴突相关肌动蛋白细胞骨架的可逆解聚。这种创新的生物药物将提供 对感觉神经元的特异性，同时利用外周神经系统的特征来消除 局部疼痛，不与中枢神经系统相互作用。 Neurocarrus已经完成了SBIR第一阶段，证明了N-001作为疼痛管理的可行性 疗法结果表明，N-001通过有效地减少机械性疼痛来管理伤害性术后疼痛。 在小鼠爪切口模型中，与布比卡因相比， 活动持续时间更长。N-001保留效力3天，而布比卡因仅保留效力6小时。N-001是 也被评估为神经阻滞模型中的麻醉剂，其中它也显示出显著增加的 与布比卡因相比，术后疼痛管理的持续时间。验证了N-001的作用机制 在体内，表明它与CGRP阳性感觉神经元而不是运动神经元共定位，并且可以 使用ADP-核糖基化肌动蛋白作为F至G肌动蛋白神经元含量的量度进行定量监测。初步 ADME、毒理学和免疫原性试验显示，如果 药代动力学信息，以及仅在多次给药后的非中和抗药抗体形成。这些 数据建立了使用N-001作为术后疼痛治疗的具体指标，从而加强了 N-001用于临床疼痛管理的潜力。 在SBIR第二阶段项目中，Neurocarrus将优化N-001的生产和配方， 制定生产标准和控制措施，以获得GLP级（药物非临床研究质量管理规范）N- 001. GLP级N-001将用于进行关键临床前研究，以证明其体内安全性， 两种临床前动物模型（C57 BL/6小鼠和比格犬）。GLP药物也将用于疗效研究 作为使用小鼠远端胫骨肢骨折模型的外周关节手术后疼痛的治疗。的 该项目的完成将支持研究性新药灌装（IND），使未来的临床试验。

PROJECT SUMMARY There is an urgent need for new approaches to treat acute human pain without the risk of Substance Use Disorders (SUDs). The most effective approved pain pharmaceuticals, including narcotics and anesthetics, are not neuron-specific and consequently suffer from off-target effects like addiction, inhibition of motor neurons, and destruction of the surrounding tissues. When inflammation occurs, actin polymerization occurs in sensory neurons, leading to the sensitization of purinergic receptors and abnormal pain behaviors. Targeted actin remodeling could be an effective approach to reduce acute nociceptive pain, but there are no small-molecule inhibitors with adequate specificity for sensory neurons that correctly modulate the cytoskeleton. Neurocarrus proposes a new therapeutic approach for nociceptive pain based on an innovative engineered protein called N- 001 that selectively targets sensory neurons and acts only at the intra-cellular level inducing limited and reversible depolymerization of the axon-associated actin cytoskeleton. This innovative biologic drug will provide specificity towards sensory neurons while leveraging the features of the peripheral nervous system to eliminate pain locally without interacting with the central nervous system. Neurocarrus has completed an SBIR Phase I that has proven the feasibility of N-001 as a pain management therapy. Results show that N-001 managed nociceptive post operative pain by efficiently reducing mechanical allodynia and gait dysfunction in a mouse paw incision model relative to bupivacaine but with a significantly longer duration of activity. N-001 retained efficacy for 3 days relative to only 6 hours for bupivacaine. N-001 was also assessed as an anesthetic agent in a nerve block model where it also showed a significantly increased duration of post operative pain management relative to bupivacaine. N-001’s mechanism of action was validated in vivo, showing that it co-localizes with CGRP positive sensory neurons not motor neurons, and can be quantitatively monitored using ADP-ribosylated actin as a measure of F to G actin neuronal content. Preliminary ADME, toxicology and immunogenicity assays showed no adverse effects on organ function, provided pharmacokinetic information, and non-neutralizing antidrug antibody formation only after multiple doses. These data establish specific metrics for the use of N-001 as a post operative pain therapeutic thereby strengthening the potential for use of N-001 in clinical pain management. In this SBIR Phase II project, Neurocarrus will optimize the production and formulation of N-001 as well as the development of manufacturing standards and controls for obtaining GLP-grade (Good Laboratory Practice) N- 001. GLP-grade N-001 will be used to perform pivotal pre-clinical studies to demonstrate its in vivo safety using two preclinical animal models (C57BL/6 mice and Beagle dogs). GLP drug will also be used for efficacy studies as a treatment for pain after peripheral joint surgery using a mouse distal tibial limb fracture model. The completion of this project will support an investigational new drug filling (IND) enabling future clinical trials.