Chemistry/Biochemistry/Cell Biology/Pharmacology Core

化学/生物化学/细胞生物学/药理学核心

基本信息

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
项目类别：
财政年份：
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

项目摘要

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足够的量化以满足四个项目所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，包括 125I- 放射性标记的配体； (3) 测试“成功的共价配体”不可逆结合的能力至CB1和CB2； (3)配体的生化和药理学表征； (5) 测试配体能够充当 anandamide 酰胺酶 (ANAase) 的底物或抑制剂； (6) 的表位标记的 CB1 和 CB2 构建体的开发及其在哺乳动物细胞中的表达；和（7） CB1和CB2受体的抗体的制备。放射性标记和非放射性标记化合物在核心 B 的支持下生产的产品也将提供给本报告中确定的其他实验室。其合作不需花费赠款的计划项目。所有成功的配体，无论是可逆的还是不可逆的，都将进行生化和药理学表征通过确定它们作为激动剂或拮抗剂的活性。在生化方面，将测试类似物：(a) 它们对 GTPyS 结合和腺苷酸环化酶的影响以及 (b) 它们在膜中的分配特性。药理学表征将包括体内生物测定，以测量行为测试的四联体大麻素药理学特征，包括运动、低温、镇痛和不动。还将检查一组代表性化合物诱导耐受性的能力。