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: 8261966
• 负责人: Richard Irving Duclos
• 金额: $ 39.21万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2014-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8261966
• 关键词: 2-arachidonylglycerol; Active Sites; 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; Cell Culture Techniques; Cellular biology; Characteristics; Chemicals; Chemistry; Collaborations; Computer Simulation; Core Facility; Development; Eating; Endocannabinoids; 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; 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; pharmacokinetic characteristic; programs; radioligand; radiotracer; receptor; receptor binding; research study; response; therapeutic target

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.

该核心基金将作为本项目三个项目（1、2、3）的技术和科学支持单位。 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的类似物， （二）研究所的研究工作需要足够的数量; 测试所有新的类似物对CB 1和CB 2大麻素受体的亲和力，包括125 I- 放射性标记的配体;（3）测试“成功的共价配体”不可逆结合的能力 CB 1和CB 2;（3）配体的生物化学和药理学表征;（5） 配体，因为它们能够充当花生四烯酸酰胺酶（ANAase）的底物或抑制剂;（6）所述配体， 开发表位标记的CB 1和CB 2构建体及其在哺乳动物细胞中的表达;和（7） CB 1和CB 2受体抗体的制备。放射性标记和非放射性标记化合物 在核心B的赞助下产生的，也将提供给本报告中确定的其他实验室。 计划项目，其合作是在没有成本的赠款。 所有成功的配体，可逆的和不可逆的，将进行生物化学和生物化学表征。 通过测定它们作为激动剂或拮抗剂的活性。在生物化学方面，将对类似物进行以下测试：（a） 它们对GTP γ S结合和腺苷酸环化酶的影响和（B）它们在膜中的分配特性。 药理学特征将包括体内生物测定来测量行为测试的四分体 大麻素药理学的特征，包括运动，低温，抗伤害感受和 不动还将检查一组代表性化合物诱导耐受性的能力。