A Novel Assay to Improve Translation in Analgesic Drug Development

项目摘要 这一新的R61/R33应用响应了NOT-NS-22-095：“疼痛相关药物的开发和验证” 促进非成瘾性镇痛药发现的模型和终点。2017年，美国国立卫生研究院（National Institutes of Health） 启动了HEAL计划，该计划的一个主要目标是开发新的，非成瘾性的 止痛药镇痛药物的发现依赖于早期疼痛和镇痛的临床前模型 候选疗法的评估，但传统的模型，依赖于反射退缩反应引起的 热刺激或机械刺激在面孔和预测效度上有显著缺陷。为了弥补这些不足， 我们花了十多年的时间开发和完善新的行为终点， 正常行为的抑郁症。疼痛-抑郁行为的测定提高了表面效度，因为行为 抑郁和功能障碍是疼痛诊断和恢复正常功能的主要标志 行为通常是疼痛治疗的主要目标。这些测定还显著提高了预测有效性 通过消除产生运动障碍的药物的假阳性效应。本申请寻求资助 为了进一步验证用于小鼠的新程序，该程序测量了 复杂环境作为一种新型的测定疼痛抑郁行为。初步数据显示， 该过程激发了高的和可复制的基线行为，通过急性内脏疼痛刺激的可靠抑郁， 临床有效的镇痛药作为阳性对照，但两个突出类别的镇痛药不能逆转疼痛效应 作为阴性对照。该程序还具有促进成果可复制性的属性， 实验设计的有效性、性别作为生物学变量的评估以及数据的定量客观性 收藏.拟议的进一步验证研究将分两个阶段进行。在第一个R61阶段（第1年）， 我们建议进一步研究，以达到三个里程碑。(1)进一步验证我们的程序与其他积极的 和阴性对照组进行对比。(2)评价药物作用的选择性，以恢复 急性疼痛引起的行为抑郁与非疼痛治疗（锂）引起的行为抑郁的比较 氯化物）。(3)评估行为终点对更持续的皮肤和内脏刺激模型的敏感性 炎性疼痛（足底完全弗氏佐剂，Ipl CFA;手术后腹部切口，AI）。在 在第二个R33阶段（第2-3年），我们建议与弗吉尼亚大学和维克森林大学的同事合作 大学以三种方式扩大和扩展该模式。(1)从ICR远交小鼠扩展到近交系 通常用于产生基因改变的小鼠（C57 BL/6 J）。(2)扩展到另外三种疾病模型， 慢性疼痛（使用备用神经损伤的单神经病疼痛，SNI;肠易激综合征，IBD;镰状细胞 贫血，SCA）。(3)评估在合作学术机构（维克森林）独立复制关键成果的情况 大学）。该项目的成功完成将为广泛的 使用这种方法来努力发现非阿片类非成瘾性镇痛药物。

Project Summary This new R61/R33 application responds to NOT-NS-22-095: “Development and Validation of Pain-Related Models and Endpoints to Facilitate Non-Addictive Analgesic Discovery.” In 2017, the National Institutes of Health launched the HEAL initiative, and one major goal of this initiative has been to develop new, non-addicting analgesics. Analgesic drug discovery depends on preclinical models of pain and analgesia for early-stage evaluation of candidate therapeutics, but conventional models that rely on reflex-withdrawal responses elicited by thermal or mechanical stimuli have significant deficits in face and predictive validity. To address these deficits, we have spent more than a decade developing and refining novel behavioral endpoints that focus on pain-related depression of normal behavior. Assays of pain-depresse behavior improve face validity because behavioral depression and functional impairment are cardinal signs of pain diagnosis, and restoration of normal function and behavior is often a major goal of pain treatment. These assays also significantly improve predictive validity by eliminating false-positive effects with drugs that produce motor impairment. This application seeks funding for further validation of a new procedure for use in mice that measures unconditioned locomotor activity in a complex environment as a new type of assay for pain-depressed behavior. Preliminary data suggest that the procedure elicits high and replicable baseline behavior, reliable depression by an acute visceral pain stimulus, and reversal of pain effects by clinically effective analgesics as positive controls but not by two prominent classes of non-analgesics as negative controls. The procedure also has attributes to promote replicability of results, efficiency of experimental design, assessment of sex as a biological variable, and quantitative objectivity of data collection. Proposed studies for further validation would proceed in two phases. In the first R61 phase (Year 1), we propose further studies to meet three milestones. (1) Further validate our procedure with additional positive and negative controls against our visceral acute pain model. (2) Evaluate selectivity of drug effects to restore behavioral depression by acute pain in comparison to behavioral depression by a non-pain treatment (lithium chloride). (3) Evaluate sensitivity of behavioral endpoints to models of more sustained cutaneous and visceral inflammatory pain (intraplantar complete Freund’s adjuvant, Ipl CFA; post-surgical abdominal-incision, AI). In the second R33 phase (Years 2-3), we propose to collaborate with colleagues at VCU and Wake Forest University to scale up and extend the model in three ways. (1) Extend from ICR outbred mice to an inbred strain commonly used to generate gene-altered mice (C57BL/6J). (2) Extend to three additional disease models of chronic pain (mononeuropathy pain using spared nerve injury, SNI; irritable bowel disease, IBD; sickle-cell anemia, SCA). (3) Evaluate independent replication of key results at a partner academic institution (Wake Forest University). Successful completion of the project would provide a comprehensive empirical foundation for broad use of this procedure in efforts to discover non-opioid non-addicting analgesic drugs.