Modulation of pain mechanisms by cannabis-derived phytochemicals.

大麻衍生的植物化学物质调节疼痛机制。

• 批准号: 10530646
• 负责人: Kenneth Mackie
• 金额: $ 63.82万
• 依托单位: QUEEN'S MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10530646
• 关键词: Abate; Acceleration; Acute; Address; Affect; Agonist; American; Analgesics; Animal Model; Anti Inflammatory Analgesics; Anti-Inflammatory Agents; Autoimmune Diseases; Binding Proteins; Biochemical; Biological; Biological Assay; Biology; Biophysics; CNR1 gene; CNR2 gene; Calcium Signaling; Cannabinoids; Cannabis; Cannabis sativa plant; Cell Line; Cells; Cellular Assay; Cessation of life; Chronic; Data; Drug Kinetics; Electrophysiology (science); Endocannabinoids; Evaluation; Excretory function; Flavonoids; Freund' s Adjuvant; G-Protein-Coupled Receptors; GPR55 receptor; GTP-Binding Proteins; Genetic; Human; Immune; Immunoassay; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Ion Channel; Lignans; Link; Macrophage; Maintenance; Malignant Neoplasms; Measures; Mediating; Membrane Potentials; Metabolism; Microglia; Microsomes; Molecular; Molecular Target; Natural Products Chemistry; Nature; Neurologic Dysfunctions; Neuropathy; Nociception; Opioid Analgesics; Paclitaxel; Pain; Pathway interactions; Pharmacology; Phytochemical; Plants; Plasma Proteins; Potassium Channel; Property; Public Health; Research; Signal Pathway; Signal Transduction; Small Interfering RNA; Solubility; System; T-Lymphocyte; TRP channel; TRPV1 gene; Terpenes; Therapeutic; Western Blotting; Work; absorption; animal pain; aqueous; cellular imaging; chronic pain; combinatorial; cytokine; economic cost; efficacy evaluation; experimental study; high throughput screening; human model; in vivo; in vivo Model; inflammatory pain; inhibitor; knowledge base; mast cell; monocyte; mouse model; musculoskeletal injury; nervous system disorder; neutrophil; opioid epidemic; opioid overdose; opioid use; pain model; painful neuropathy; patch clamp; pharmacologic; receptor; screening; side effect; sulfated glycoprotein 2

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.

项目总结/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- 影响了约1亿美国人。过度依赖阿片类镇痛剂导致了全国公共卫生危机， 阿片类药物过量在2017年夺去了47，000多人的生命，现在是该国可避免死亡的主要原因。 大麻植物具有镇痛和抗炎特性，因为它含有丰富的大麻素，萜烯， 木脂素和类黄酮。然而，对众多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）--受到限制。我们已经组装了一个互补的 和跨学科的团队，结合了分子和细胞信号传导，离子通道生物学，自然 产品化学和分子药理学以及内源性大麻素生物学的各个方面。我们初步的高... 通量筛选（HTS）生物测定已经鉴定了几种大麻素，其抑制免疫细胞中的钙信号传导。 细胞，因此可以减少炎症和相关的疼痛。本文提出的工作结果将确定 Cannabis sativa中含有的抗炎分子，并描述了它们参与的作用机制。 我们假设大麻中的特定植物化学物质抑制细胞Ca 2+信号传导和随后的促钙释放。 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， 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， 免疫细胞Ca 2+信号传导和细胞因子释放，从而改善炎性疼痛。 我们建议对大麻植物化学物质的个体和环境影响进行药理学分析， 5种特定促炎人类免疫细胞中的Ca 2+信号传导（Aim 1A）。我们将确定细胞和分子 这些免疫细胞中活性大麻植物化学物质参与的Ca 2+动员机制（Aim 1B）; 大麻植物化学物质对伤害性疼痛、炎症性疼痛和神经性疼痛的既定分子靶点（特异性 TRP通道和G蛋白），使用异源表达系统、HTS生物测定和单细胞电生理学（Aim 1C）。在目的2中，将使用大麻素的体内小鼠模型评估活性大麻素和组合的镇痛性质。 炎性和神经性疼痛。在这里，我们将首先确定体外“吸收、分布、代谢和代谢"。 所述大麻素的“排泄”（ADME）性质（目的2A）和体内药代动力学（目的2B）。我们将评估 在完全弗氏佐剂（CFA）诱导的炎症和紫杉醇介导的 中毒性神经性疼痛（Aim 2C）。 总之，这些研究将建立一个全面的和机械的知识基础，关于疗效，效力和 大麻衍生的植物化学物质作为抗炎镇痛药的适用性，并可能有助于改善 阿片类药物的流行