Chemistry/Biochemistry/Cell Biology/Pharmacology Core

这一核心设施将作为三个项目(1、2、3)的技术和科学支助单位 计划项目。其主要目标包括：(1)生产放射性标记和非放射性标记类似物 足够的数量，以满足参与四个项目的研究实验室的需要； 测试所有新类似物对CB1和CB2大麻素受体的亲和力，包括125I- 放射性标记配体；(3)“成功的共价配体”不可逆结合能力的测试 对CB1和CB2的作用；(3)配体的生化和药理特性；(5)对CB1和CB2的检测 作为氨基酰胺酶(ANAase)底物或抑制剂的配体； 标记表位的CB1和CB2构建及其在哺乳动物细胞中的表达 CB1和CB2受体抗体的制备。放射性标记化合物和非放射性标记化合物 在核心B的支持下生产的产品也将提供给本 合作不向拨款支付费用的计划项目。 所有成功的配体，无论是可逆的还是不可逆的，都将经过生化和药理学的表征。 通过确定它们作为激动剂或拮抗剂的活性。在生物化学方面，这些类似物将被测试：(A) 它们对GTPyS结合和腺苷环化酶的影响以及(B)它们在膜中的分配特性。 药理特性将包括体内生物测试，以测量行为测试的四元组 大麻素类药物的特点，包括运动、低温、抗伤害和 一动不动。还将检查一组具有代表性的化合物是否具有诱导耐受性的能力。

This Core Facility will serve as a technical and scientific support unit for the three projects (1, 2, 3) of this Program Project. Its major goals involve: (1) the production of radiolabeled and nonradiolabeled analogs in sufficient quantifies to address the needs of the research laboratories involved in the four projects; (2) the testing of all new analogs for their affinities for the CB1 and CB2 cannabinoid receptors, including the 125I- radiolabeled ligands; (3) the testing of the "successful covalent ligands" for their abilities to irreversibly bind to CB1 and CB2; (3) the biochemical and pharmacological characterization of the ligands; (5) the testing of ligands for their abilities to act as substrates or inhibitors of anandamide amidase (ANAase); (6) the development of epitope tagged CB1 and CB2 constructs and their expression in mammalian cells; and (7) the preparation of antibodies for the CB1 and CB2 receptors. Radiolabeled and non-radiolabeled compounds produced under the auspices of Core B will also be made available to other laboratories identified in this program project whose collaboration is at not cost to the grant. All successful ligands, reversible and irreversible, will be characterized biochemically andpharmacologically by determining their activities as agonists or antagonists. Biochemically, the analogs will be tested for: (a) their effects on GTPyS binding and adenylate cyclase and (b) their partitioning properties in the membrane. The pharmacological characterization will include in vivo bioassaysto measure the tetrad of behavioraltests characteristic of cannabinoid pharmacology including locomotor, hypothermia, antinociception and immobility. A representative group of compounds will also be examined for their ability to induce tolerance.