Chemistry/Biochemistry/Cell Biology/Pharmacology Core

化学/生物化学/细胞生物学/药理学核心

• 批准号: 7222484
• 负责人: Richard Irving Duclos
• 金额: $ 38.31万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-15 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7222484
• 关键词: 2-arachidonylglycerol; Address; Adenylate Cyclase; Affinity; Agonist; Amino Acids; Antibodies; Area; Behavioral; Behavioral Assay; Binding; Binding Sites; Biochemical; Biochemistry; Blood - brain barrier anatomy; Brain; CNR1 gene; CNR2 gene; Cannabinoids; Cellular biology; Characteristics; Chemicals; Chemistry; Class; Collaborations; Computer Simulation; Core Facility; Cultured Cells; Development; Drug Kinetics; Eating; Epitopes; Generations; Goals; Grant; High Pressure Liquid Chromatography; Human; I125 isotope; Image; Inhibitory Concentration 50; Intestinal Absorption; Label; Laboratories; Laboratory Research; Ligands; Mammalian Cell; Measures; Membrane; Metabolic; Microsomes; Monoacylglycerol Lipases; Motor Activity; Mus; Nature; Numbers; Octanols; Oocytes; Organ; Partition Coefficient; Penetration; Pharmacology; Phase; Plasma Proteins; Preparation; Principal Investigator; Production; Property; Protein Binding; Radiolabeled; Rattus; Research Personnel; Screening procedure; Signal Transduction; Site; Structure; Surface; Testing; Tritium; Water; analog; anandamide; arachidonoylethanolamide synthase; base; cannabinoid receptor; cost; cyclooxygenase 2; design; drug metabolism; fatty acid amide hydrolase; in vitro Assay; in vivo; inhibitor/antagonist; mutant; natural hypothermia; novel; programs; radioligand; radiotracer; receptor; receptor binding; research study; response

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.

该核心设施将作为这三个项目（1、2、3）的技术和科学支持部门 计划项目。它的主要目标涉及：（1）在 足够的量化以满足四个项目涉及的研究实验室的需求； （2） 测试所有新类似物对CB1和CB2大麻素受体的亲和力的测试，包括125I- 放射性标记的配体； （3）测试“成功的共价配体”，以使其能力不可逆地绑定 到CB1和CB2； （3）配体的生化和药理学表征； （5）测试 配体的能力充当anandamide amidase（Anaase）的底物或抑制剂的能力； （6） 表位的开发标记为CB1和CB2构建体及其在哺乳动物细胞中的表达； （7） 制备CB1和CB2受体的抗体。放射性标记和非二醇标记化合物 在核心B的主持下生产的其他实验室也将提供给其他实验室 计划项目的合作不花钱支付赠款。 所有成功的配体，可逆和不可逆的，都将在生化和肢体学上被描述 通过确定他们作为激动剂或对手的活动。从生化上，将对类似物进行测试：（a） 它们对GTPY的结合和腺苷酸环化酶的影响以及（b）它们在膜上的分配特性。 药理学表征将包括体内生物测定法测量行为测试的四分法 大麻素药理学的特征，包括运动，体温过低，抗伤害感受和 不动。还将检查一组代表性的化合物，以诱发耐受性。