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: 10152004
• 负责人: Kenneth Mackie
• 金额: $ 66.34万
• 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/10152004
• 关键词: Abate; 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 Studies; Excretory function; Flavonoids; Freund' s Adjuvant; G-Protein-Coupled Receptors; GPR55 receptor; GTP-Binding Proteins; Genetic; Human; Immune; Immunoassay; Immunoblotting; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Ion Channel; Lignans; Link; Maintenance; Malignant Neoplasms; Measures; Mediating; Membrane Potentials; Metabolism; Microglia; Microsomes; Molecular; Molecular Target; Natural Products; 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; Work; absorption; animal pain; aqueous; cellular imaging; chronic pain; combinatorial; cytokine; economic cost; experimental study; high throughput screening; human model; in vivo; in vivo Model; inflammatory pain; inhibitor/antagonist; knowledge base; macrophage; mast cell; monocyte; mouse model; musculoskeletal injury; nervous system disorder; neutrophil; opioid epidemic; opioid overdose; opioid use; pain model; painful neuropathy; patch clamp; 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.

项目摘要/摘要 慢性疼痛 - 通常是由肌肉骨骼损伤，神经功能障碍，癌症或自身免疫性疾病引起的 - 影响约1亿美国人。对阿片类镇痛药的过度依赖导致国家公共卫生危机 阿片类药物过量在2017年夺去了47,000多人的生命，现在是全国可避免死亡的主要原因。 大麻植物具有镇痛和抗炎特性，这是由于其大麻素含量，萜烯， 木质杆和类黄酮。但是，研究众多生物活性的生物学效应和分子机制 植物化学物质 - 单独或组合（随行效应）是有限的。我们已经组装了一个完整的 和跨学科团队，结合了分子和细胞信号传导，离子通道生物学，自然的专业知识 产品化学，分子药理学以及内源性大麻素生物学的各个方面。我们的初步高级 吞吐量筛选（HTS）生物测定已鉴定出几种抑制免疫中钙信号的大麻素 细胞，因此可能减少感染和相关的疼痛。这里提出的工作的结果将确定 大麻sativa中包含的抗炎分子，表征了它们所参与的作用机制。 我们假设大麻中的特定生理化学物质抑制了细胞Ca2+信号传导，并随后释放促疾病 免疫细胞中炎性细胞因子，导致炎症性疼痛。我们进一步假设组合 大麻物理协同抑制某些离子通道和G蛋白偶联受体 免疫细胞CA2+信号传导和细胞因子释放，从而改善炎症性疼痛。 我们建议对大麻生理学对个体和entoiourage效应进行药物分析 5个特定促炎性人类免疫力（AIM 1A）中的Ca2+信号传导。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CA2+动员机制（AIM 1B）；和个人资料 大麻对伤害性，炎症性和神经性疼痛的既定分子靶标（特定 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AIM 2中，将使用活性大麻素的镇痛特性以及使用体内小鼠模型的组合 炎症和神经性疼痛。在这里，我们将首先确定体外“吸收，分布，代谢和 排泄”（ADME）特性（AIM 2A）和体内药代动力学（AIM 2B）上述大麻素。然后，我们将评估 完整的弗雷德佐剂（CFA）诱导的注射和紫杉醇介导的最有利的大麻素（S） 有毒神经性疼痛（AIM 2C）。 这些研究将共同​​创建有关效率，效能和的全面和机械知识基础 大麻衍生的物理化学物质作为抗炎镇痛药的适用性，可能有助于改善 当前的阿片类药物流行。