An all-optical electrophysiology platform for the discovery of pain therapeutics

项目摘要 慢性疼痛在美国影响着1亿多成年人，治疗起来很有挑战性。 目前的治疗方法包括阿片类药物和非类固醇炎药。然而，它的功效 这些药物在慢性治疗中受到剂量限制毒性和长期阿片类药物的限制。 使用可能会导致依赖。尽管明确的、未得到满足的医疗需求和重要的研究 活性方面，基于新的、非阿片类药物机制的针对疼痛的药物很少出现在 过去十年。Q-State已经创造了一种新型的全光平台(Optopatch)，它使用Engineering 光遗传蛋白质和定制显微镜可同时刺激和记录电信号 具有高灵敏度和时间分辨率的各种细胞类型的活动。我们专注于我们的 对基因验证的疼痛靶点SCN9A(Nav1.7)的努力，SCN9A(Nav1.7)是一种电压门控钠通道 这是在感觉神经元中传递疼痛信号所必需的。我们将应用Optopatch 技术有两种格式。1)Q状态化学库的HTS屏幕使用 异源表达的复制生理刺激性的Nav1.7通道试验。 针对其他Nav1.x通道的反筛查将以相同的分析格式执行，以 确定化合物的选择性。2)确定的抑制剂将在中等产量下进行评估 测量啮齿动物感觉神经元和人类iPS感觉神经元兴奋性的屏幕 用炎症介质致敏。这套集成的检测方法旨在 确定不同作用机制的Nav1.7抑制剂，并确定化合物的优先顺序 使用可扩展的感觉神经元功能体外模型进一步优化。这个平台将 被用作一种有效的手段来选择化合物以进行药物优化 化学和药代动力学、药物代谢和体内疗效数据。

Project Summary Chronic pain affects over 100 million adults in the United States and is challenging to treat. Current treatments include opioids and non-steroidal inflammatory agents. However, efficacy of these drugs in chronic treatment is restricted by dose limiting toxicities, and prolonged opioid use can lead to dependency. Despite the clear, unmet medical need and significant research activity, few drugs targeting pain based on novel, non-opioid mechanisms have appeared in the past decade. Q-state has created a novel all-optical platform (Optopatch) using engineered optogenetic proteins and custom microscopes to simultaneously stimulate and record electrical activity from a variety of cell types with high sensitivity and temporal resolution. We focus our efforts on a genetically validated pain target, SCN9A (Nav1.7), a voltage-gated sodium channel that is required for pain signal transmission in sensory neurons. We will apply Optopatch technology in two formats. 1) An HTS screen of the Q-State chemical library using a heterologously expressed Nav1.7 channel assay that replicates physiological spiking activity. Counterscreens against other Nav1.x channels will be performed in the same assay format to determine compound selectivity. 2) Identified inhibitors will be evaluated in medium throughput screens that measure excitability in rodent sensory and human iPS sensory neurons that have been sensitized using inflammatory mediators. This integrated set of assays is designed to identify Nav1.7 inhibitors acting by diverse working mechanisms and prioritize compounds for further optimization using scalable in vitro models of sensory neuron function. This platform will be employed as an efficient means to select compounds for optimization using medicinal chemistry and pharmacokinetic, drug metabolism and in vivo efficacy data.