Anti-inflammatory Effects of Novel Minor Cannabinoids and Terpenes on Cellular and Murine Models of HIV and HIV Proteins

总结 大约50%的HIV感染患者患有顽固性疼痛，许多人自我报告， 服用大麻以减轻症状。然而，关于大麻影响的数据仍然有限。 在HIV感染者中，cART治疗人群中。研究最多的化合物是Δ 9-THC， 以及限制治疗潜力的成瘾作用。然而，有超过120种次要的大麻素和200种萜烯 我们著名的大麻研究实验室发现，其中许多含有非精神活性物质， 治疗效果我们确定了大麻成分，减少艾滋病毒介导的炎症/星形胶质细胞增生， 细胞培养和HIV蛋白介导的小鼠内脏痛。我们假设几种次要的大麻素和 萜烯将补充cART，以改善感染性HIV引起的病毒血症、炎症和细胞毒性。 1.此外，我们预计这些化合物中的几种可以改善炎症，机械，内脏， 和由cART或HIV-1蛋白促进的神经性疼痛状态。目标1将确定重要的大麻 在体外研究了调节HIV介导的病毒血症和炎症的成分和靶点。利用我们广泛的 库的大麻素，萜烯，挥发油，我们将筛选高大麻二酚（CBD）的混合物和44纯 少量大麻素和萜烯对模拟人外周血单核细胞或小胶质细胞的作用， 感染或感染HIV-1IIIB或HIV-1BaL。将评估病毒血症、细胞因子产生和细胞毒性， 随后将使用CB 1、CB 2和GPR 55的拮抗剂检查药效学机制。 目的2将确定在体内的抗伤害性和镇痛作用的次要大麻素和萜烯， HIV-1蛋白/cART相关炎性、热、内脏和神经性疼痛的啮齿动物模型。剂量- 10 β-羟基-大麻色烯的依赖性抗炎、抗痛觉过敏和抗伤害感受作用 8-四氢大麻酚和β-香芹烯将是重点关注的纯化合物，沿着高CBD提取物 (and目标1中确定的其他抗炎先导化合物）。这些成分将在男性和女性中进行评估 表达（或不表达）HIV-1达特或gp120蛋白的转基因小鼠。小鼠将维持在 cART评估潜在的改善或与大麻素相关结果的相互作用。目标3将决定 大麻素影响的重要中枢和肠神经系统炎症机制 和萜烯类化合物。疼痛相关脑区、脊髓、背根 将评估目标2中收集的神经节、回肠、水肿和血浆的细胞因子产生。脑和回肠 将通过[35S] GTP γ S试验以及神经元形态学评估CB1或CB2 G蛋白活性， 单核细胞源性细胞和星形胶质细胞增生。这些研究将提供深入了解的机制， 大麻素和萜烯的作用，将揭示抗病毒血症，抗炎和抗伤害的潜力， 非精神活性大麻成分，并将阐明治疗艾滋病毒相关和非艾滋病毒相关， 顽固性疼痛状态。

Summary Approximately 50% of HIV-infected patients suffer from intractable pain and many individuals self-report the consumption of cannabis for alleviating their symptoms. However, data are still limited on the effects of cannabis in the HIV-infected, cART-treated population. The compound most studied is Δ9-THC, which exerts psychoactive and addictive effects that limit therapeutic potential. Yet, there are over 120 minor cannabinoids and 200 terpenes in cannabis and our renowned Marijuana Research Laboratory has found many of them hold non-psychoactive, therapeutic effects. We identified cannabis constituents that reduce HIV-mediated inflammation/astrogliosis in cell culture and HIV-protein mediated visceral pain in mice. We hypothesize that several minor cannabinoids and terpenes will complement cART to ameliorate viremia, inflammation, and cytotoxicity caused by infectious HIV- 1. Moreover, we anticipate several of these compounds to ameliorate the inflammatory, mechanical, visceral, and neuropathic pain states promoted by cART or HIV-1 proteins. Aim 1 will determine the important cannabis constituents and targets that modulate HIV-mediated viremia and inflammation in vitro. Utilizing our extensive library of cannabinoids, terpenes, and volatile oils, we will screen a high-cannabidiol (CBD) mixture and 44 pure minor cannabinoids and terpenes against human peripheral blood mononuclear cells or microglia that are mock- infected or infected with HIV-1IIIB or HIV-1BaL. Viremia, cytokine production, and cytotoxicity will be assessed and the pharmacodynamic mechanisms will be subsequently examined using antagonists to CB1, CB2, and GPR55. Aim 2 will determine the in vivo anti-nociceptive and analgesic effects of minor cannabinoids and terpenes in rodent models of HIV-1 protein/cART-related inflammatory, thermal, visceral, and neuropathic pain. Dose- dependent anti-inflammatory, anti-hyperalgesic, and anti-nociceptive effects of cannabichromene, 10β-hydroxy- 8-tetrahydrocannabinol, and β–caryophyllene will be the pure compounds of focus, along with High-CBD extract (and other anti-inflammatory leads identified in Aim 1). These constituents will be assessed in male and female transgenic mice that express (or do not express) the HIV-1 Tat or gp120 proteins. Mice will be maintained on cART to assess potential improvements or interactions with cannabinoid-related outcomes. Aim 3 will determine the important central and enteric nervous system inflammatory mechanisms that are influenced by cannabinoids and terpenes following HIV-1 protein/cART exposure. Pain-related brain regions, spinal cord, dorsal root ganglion, ileum, edema, and plasma collected in Aim 2 will be assessed for cytokine production. Brain and ileum will be assessed for CB1 or CB2 G-protein activity via [35S]GTPγS assay as well as neuron morphology and monocyte-derived cell and astrogliosis. TThese studies will provide insight into the mechanisms by which minor cannabinoids and terpenes act, will reveal the anti-viremic, anti-inflammatory, and antinociceptive potential of non-psychoactive cannabis constituents, and will elucidate therapeutics for HIV-related and non-HIV-related, intractable pain states.