Cannabinoid Acids as Antiinflammatory Agents

描述(申请人提供)：本提案的总体目标是确定没有精神活性的大麻酸调节细胞(巨噬细胞、滑膜细胞)功能的生化和分子机制，这些细胞参与类风湿关节炎患者关节组织损伤的启动和传播。要研究的大麻酸包括四氢大麻酸(AJA)和四氢大麻酚-11 OIC酸。这些化合物不会改变行为，是口服的，不会损伤胃粘膜。口服AJA是四氢大麻酚(THC)羧酸代谢物THC-11-OIC酸的合成非精神活性类似物，可以预防关节炎动物模型的关节软骨和骨损伤。初步研究表明，在体外细胞中加入大麻酸可抑制几种炎性细胞因子的基因表达，并促进细胞凋亡。人类口服AJA可减少激活单核细胞产生的IL-11β。该方案的主要假设是，大麻酸类拮抗转录因子NFkB的激活，这是由于和/或不依赖于PPARy的激活，导致细胞凋亡和NFkB靶基因(IL-1、IL-6、IL-8、COX-2)的下调。还将调查二十烷类化合物在大麻酸作用中的作用。细胞将从健康志愿者以及骨关节炎和类风湿性关节炎患者那里获得。在体外，大麻酸将被添加到细胞中。口服AJA前后从健康志愿者体内获得的细胞将在体外进行研究。实验旨在确定大麻酸是否是PPARy的真正配体，并研究大麻酸改变NFkB信号的机制。调查将包括基因导入、免疫印迹和免疫显微镜的使用。这项提案的完成将使人们更好地了解大麻酸的抗炎和免疫调节作用，并可能导致更合理地设计这些化合物，用于治疗以免疫反应紊乱、炎症和组织损伤为特征的疾病。

DESCRIPTION (provided by applicant): The overall object of this proposal is to determine biochemical and molecular mechanisms whereby cannabinoid acids without psychoactive activities modulate the function of cells (macrophages, synovial cells) which participate in initiation and propagation of joint tissue injury in patients with rheumatoid arthritis. The cannabinoid acids to be studied include ajulemic acid (AjA), and tetrahydrocannabinol-11 oic acid. These compounds do not alter behavior, are given by mouth, and do not injure gastric mucosa. Oral administration of AjA, a synthetic nonpsychoactive analog of the tetrahydrocannabinol (THC) carboxylic acid metabolite THC-11-oic acid, prevents joint cartilage and bone damage in an animal model of arthritis. Preliminary studies indicate that addition of cannabinoid acids to cells in vitro suppress gene expression of several inflammatory cytokines, and enhances apoptosis. Oral administration of AjA to humans attenuates IL-11beta production by activated monocytes. The main hypothesis of the proposal is that cannabinoid acids antagonize activation of the transcription factor NFkB due to and/or independent of activation of peroxisome proliferator activated receptor gamma (PPARy), leading to enhancement of apoptosis and downregulation of NFkB target genes (Il-1, IL-6, IL-8, COX-2). The role of eicosanoids in the actions of cannabinoid acids will be investigated also. Cells will be obtained from healthy volunteers, and patients with osteoarthritis and rheumatoid arthritis. Cannabinoid acids will be added to cells in vitro. Cells obtained from healthy volunteers before and after oral administration of AjA will be studied ex vivo. Experiments are designed to determine whether cannabinoid acids are bona fide ligands for PPARy and to investigate mechanisms by which cannabinoid acids alter NFkB signaling. Investigations will include use of gene transfection, western blotting, and immunomicroscopy. Completion of this proposal will provide a greater understanding of antiinflammatory and immunomodulatory effects of cannabinoid acids, and may lead to more rational design of these compounds for treatment of patients with diseases characterized by disordered immune responses, inflammation, and tissue injury.