Modulation of pain mechanisms by cannabis-derived phytochemicals.

项目摘要/摘要 慢性疼痛--通常由肌肉骨骼损伤、神经功能障碍、癌症或自身免疫性疾病引起- 影响到约1亿美国人。过度依赖阿片类镇痛剂已导致国家公共卫生危机，其中 2017年，阿片类药物过量已夺走4.7万多人的生命，现在已成为美国可避免的死亡的主要原因。 大麻属植物因其含有丰富的大麻类化合物、萜类化合物和 木脂素和类黄酮类。然而，对多种生物活性物质的生物学效应和分子机制的研究 植物化学物质--单独或组合(环境效应)--一直受到限制。我们已经组装了一个互补的 和跨学科团队，结合了分子和细胞信号、离子通道生物学、天然 产品化学、分子药理学以及内源性大麻生物的方方面面。我们的初步高潮- 吞吐筛选(HTS)生物检测已经确定了几种抑制免疫中钙信号的大麻素 因此可能会减少炎症和相关的疼痛。这里提出的工作结果将确定 大麻中含有的抗炎分子，并描述了它们参与的作用机制。 我们假设大麻中特定的植物化学物质抑制细胞内的钙信号和随后释放的前体- 免疫细胞中的炎性细胞因子导致炎性疼痛。我们进一步假设组合 大麻植物化学物质协同抑制某些离子通道和G蛋白偶联受体参与 免疫细胞钙信号转导和细胞因子释放，从而减轻炎症性疼痛。 我们建议对大麻植物化学物质的个人和环境效应进行药理学分析 5个特定的促炎人类免疫细胞中的钙信号(目标1A)。我们将确定细胞和分子 活性大麻植物化学物质在这些免疫细胞中启动的钙动员机制(目标1B)； 大麻植物化学物质对已建立的伤害性、炎症性和神经病理性疼痛的分子靶点(特异性 Trp通道和G蛋白)使用异源表达系统、HTS生物检测和单细胞电生理(AIM 1C)。在Aim 2中，将使用活体小鼠模型评估活性大麻素及其组合的止痛特性 炎症性和神经性疼痛。在这里，我们将首先在体外测定“吸收、分布、代谢和 上述大麻素的“排泄”(ADME)性质(目标2A)和体内药代动力学(目标2B)。然后我们将评估 最有利的大麻素(S)在完全弗氏佐剂诱导的炎症和紫杉醇介导的炎症中的作用 中毒性神经病理性疼痛(目标2C)。 总之，这些研究将建立一个关于疗效、效力和 大麻衍生植物化学物质作为抗炎镇痛剂的适宜性，并可能有助于改善 目前流行的阿片类药物。

PROJECT SUMMARY/ABSTRACT Chronic pain – often arising from musculoskeletal injury, neurological dysfunction, cancer, or autoimmune disorders – affects ~100 million Americans. Overreliance on opioid analgesics has resulted in a national public health crisis in which opioid overdoses have claimed over 47,000 lives in 2017 and are now the leading cause of avoidable deaths in the nation. The Cannabis plant has analgesic and anti‐inflammatory properties owing to its rich content of cannabinoids, terpenes, lignans, and flavonoids. However, research on the biological effects and molecular mechanisms of the numerous bioactive phytochemicals – alone or in combination (entourage effect) – has been limited. We have assembled a complementary and interdisciplinary team that combines expertises in molecular and cellular signaling, ion channel biology, natural products chemistry, and molecular pharmacology as well as all aspects of endocannabinoid biology. Our preliminary high‐ throughput screening (HTS) bioassays have identified several cannabinoids that inhibit calcium signaling in immune cells and may therefore reduce inflammation and the associated pain. Results from the work proposed here will identify the anti‐inflammatory molecules contained in Cannabis sativa and characterize the mechanisms of action they engage. We hypothesize that specific phytochemicals in Cannabis suppress cellular Ca2+ signaling and subsequent release of pro‐ inflammatory cytokines in immunocytes that contribute to inflammatory pain. We further hypothesize that combinations of Cannabis phytochemicals synergistically inhibit certain ion channels and G protein‐coupled receptors involved in immunocyte Ca2+ signaling and cytokine release, thereby ameliorating inflammatory pain. We propose to perform pharmacological profiling of individual and entourage effects of Cannabis phytochemicals on Ca2+ signaling in 5 specific pro‐inflammatory human immune cells (Aim 1A). We will determine the cellular and molecular Ca2+ mobilizing mechanisms engaged by active Cannabis phytochemicals in these immune cells (Aim 1B); and profile Cannabis phytochemicals on established molecular targets of nociceptive, inflammatory and neuropathic pain (specific TRP channels and G‐proteins) using heterologous expression systems, HTS bioassays and single cell electrophysiology (Aim 1C). In Aim 2 will assess analgesic properties of active cannabinoids and combinations using in vivo mouse models of inflammatory and neuropathic pain. Here, we will first determine in vitro “Absorption, Distribution, Metabolism, and Excretion” (ADME) properties (Aim 2A) and in vivo pharmacokinetics (Aim 2B) of said cannabinoids. We will then assess the most favorable cannabinoid(s) in Complete Freund's Adjuvant (CFA)‐induced inflammation and paclitaxel‐mediated toxic neuropathic pain (Aim 2C). Together, these studies will create a comprehensive and mechanistic knowledge base about the efficacy, potency and suitability of Cannabis‐derived phytochemicals as anti‐inflammatory analgesics and may contribute to ameliorating the current opioid epidemic.