Cannabimimetic Ligands and Endocannabinoid Active Sites

该项目作为配体/药物开发的组成部分，全面合作努力 在程序拒绝(PP)的支持下研究内源性大麻素的作用部位。高亲和力 将利用共价和非共价配体来获得关于活性中心的详细结构信息 CB1和CB2大麻素受体(CBRs)。提高我们开发高CB1配体的能力 或CB2选择性，我们还将测试我们的新型配体与KEY相互作用的能力 参与内源性大麻素失活的内源性大麻素靶标，包括两种水解酶， 脂肪酸酰胺水解酶(FAAH)和单酰甘油连接酶(MGL)--氧化酶 环氧合酶-2(COX-2)和双胺转运系统(ANT)。这些配体将被用来： (A)探测CB1和CB2结合部位；(B)活体成像哺乳动物大脑中的CB1受体(成像 (C)探讨不同类别配体的药效要求 用于CB1和CB2的脱敏以及受体的激活、失活和二聚化。新的 化合物代表了结构上不同类别的大麻能药物，包括经典的和非 经典的大麻素、氨基烷基吲哚、烷基酰胺和2-花生四烯基甘油类似物和吡唑 CB1和CB2拮抗剂。配体被设计成可光活化或亲电的不可逆探针。 或作为高亲和力可逆探针。将在杆状病毒细胞培养中进行共价受体标记 表达表位标记的CB1和CB2的制备。受体的表达、纯化和鉴定 鉴定将利用亲和层析和质谱学方法。关于配基的信息- 受体的相互作用将用于非阿片类、非成瘾的中枢和外周的发育 止痛药，以及旨在对抗大麻和其他滥用药物的不良影响的药物。 这种药物旨在解决吸毒成瘾的流行问题，将具有很大的医学价值和 社会效益。

This project serves as the ligand/drug development component of a comprehensive collaborative effort to study the endocannabinoid sites of action under the auspices of a Program Reject (PP). High affinity covalent and non-covalent ligands will be utilized to obtain detailed structural information on the active sites of the CB1 and CB2 cannabinoid receptors (CBRs). To enhance our ability to develop ligands with high CB1 or CB2 selectivities, we shall also test our novel ligands for their abilities to interact with key endocannabinoid targets involved in endocannabinoid deactivation including the two hydrolytic enzymes, Fatty Acid Amide Hytlrolase (FAAH) and Monacyl Glycerol Ligase (MGL), the oxidative enzyme Cyclooxygenase-2 (COX-2), and the Anandamide Transport system (ANT). The ligands will be utilized to: (a) probe the CB1 and CB2 binding sites; (b) in vivo image the CB1 receptor in mammalian brains (imaging of FAAH will be a later goal); (c) explore the pharmacophoric requirements within different classes of ligands for CB1 and CB2 desensitization as well as receptor activation, deactivation and dimerization. The new compounds represent structurally diverse classes of cannabinergic agents, including classical and non- classical cannabinoids, aminoalkylindoles, anandamide and 2-arachidonoyl glycerol analogs and pyrazole CB1 and CB2 antagonists. The ligands are designed as photoactivatable or electrophilic, irreversible probes or as high affinity reversible probes. Covalent receptor labeling will be carried out in baculovirus cell culture preparations expressing epitope-tagged CB1 and CB2. Receptor expression, purification and characterization will utilize affinity chromatographic and mass spectroscopic methods. Information on ligand- receptor interactions will be used for the development of non-opioid, non-addictive central and peripheral analgesics, as well as medications designed to combat the ill effects of cannabis and other drugs of abuse. Such medications aim at addressing the drug addiction epidemic and will be of great medical value and social benefit.