In vitro and in vivo characterization of CB1 allosteric modulators

CB1变构调节剂的体外和体内表征

基本信息

批准号：
10431876
负责人：
Thomas F Gamage
金额：
$ 0.9万
依托单位：
RESEARCH TRIANGLE INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-15 至 2023-08-11
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10431876
关键词：
2-arachidonylglycerol Affect Affinity Agonist Antibodies Attenuated Award Behavior Behavioral Behavioral Assay Behavioral Model Binding Biological Biological Assay Budgets CNR1 gene CNR2 gene Cannabinoids Cells Chemicals Clinical Clinical Data Complex Coupling Data Development Disease Dronabinol Drug Addiction Drug Targeting Drug abuse Educational workshop Endocannabinoids Enhancers Exhibits Feeling suicidal Funding GTP Binding GTP-Binding Proteins Goals In Vitro K-Series Research Career Programs Laboratories Learning Ligand Binding Ligands Mental Depression Mentors Metabolic syndrome Modeling Modification Molecular Molecular Biology Molecular Conformation Mus Neuraxis New England Obesity Opioid Pain Parents Pharmaceutical Preparations Pharmacology Pharmacology Study Pharmacotherapy Physiological Pre-Clinical Model Principal Investigator Receptor Signaling Reporting Research SR141716 Self Administration Signal Pathway Signal Transduction Signaling Protein Site Structure Structure-Activity Relationship Techniques Testing Therapeutic Training addiction analog anandamide antagonist antagonist G base cannabinoid receptor career development clinically relevant experimental study improved in vivo induced hypothermia novel opioid abuse opioid withdrawal pre-clinical programs radioligand receptor response responsible research conduct rimonabant scaffold side effect small molecule stoichiometry success synthetic cannabinoid targeted treatment theories therapeutic target vector

项目摘要

The cannabinoid type-1 (CB1) receptor is a promising pharmacotherapeutic target for many diseases as evidenced by a large body of preclinical and clinical data showing efficacy treating drug dependence/addiction, pain, obesity/metabolic syndrome, and others. Unfortunately drugs developed that target the CB1 receptor either directly or indirectly have had limited success, i.e. dronabinol produces undesired psychoactivity, rimonabant produces depression/suicidal ideation, and PF-04457845 did not demonstrate efficacy. Important considerations in the therapeutic vs. nontherapeutic effects of CB1 activation are the various signaling pathways that contribute to either of these. Allosteric modulation provides an additional vector through which the CB1 receptor can be manipulated for therapeutic gain. This study proposes to 1) examine novel structural analogs of established CB1 allosteric modulators for their ability to alter orthosteric ligand binding, function, and signaling bias; 2) characterize G protein subtype-dependent coupling by orthosteric and allosteric ligands in N18TG2 cells which provide physiologically relevant receptor/G protein stoichiometry; and 3) assess the effects of CB1 allosteric modulators in assays of cannabinoid activity, opioid withdrawal and self-administration for determination of efficacy in therapeutically relevant models. My goals for this career development award are to build on my laboratory’s capabilities and prepare for independence as a principal investigator in the behavioral and molecular pharmacology of drug abuse. My laboratory currently has the capabilities to carry out the proposed studies but I require additional training in molecular pharmacology to learn G protein antibody capture scintillation proximity assay to complete Aim 2 which I will learn in Dr. Allyn Howlett’s laboratory. I will also learn complementary techniques in molecular biology in a workshop from New England BioLabs. I will also benefit from guidance in receptor theory, application of allosteric models and quantification of signaling bias which I will receive from my co-mentor Dr. Terry Kenakin. I also require training in self-administration for Aim 3 experiments which I will learn in Dr. Jenny Wiley’s laboratory. In addition to technical training, I require career development in order prepare for research independence. Under this award I will receive training in grantsmanship, laboratory management, budget management, and responsible conduct of research. These career development activities will include one-on-one training with co-mentors as well as workshops to provide me with necessary training to successfully compete for R01 funding and develop a highly productive and efficient independent research program.

1 型大麻素 (CB1) 受体是许多疾病的有前途的药物治疗靶点，例如大量临床前和临床数据证明了治疗药物依赖/成瘾的功效，不幸的是，针对 CB1 受体的药物已开发出来。直接或间接取得的成功有限，即屈大麻酚会产生不良的精神活性，利莫那班产生抑郁/自杀意念，并且 PF-04457845 没有表现出功效。 CB1 激活的治疗效果与非治疗效果的影响因素是各种信号通路变构调节为 CB1 受体提供了一个额外的载体。本研究建议 1) 检查已建立的 CB1 的新结构类似物。变构调节剂能够改变正位配体结合、功能和信号偏向；2) 表征 N18TG2 细胞中正构和变构配体的 G 蛋白亚型依赖性偶联，提供生理相关的受体/G 蛋白化学计量；以及 3) 评估 CB1 变构的影响大麻素活性测定、阿片类药物戒断和自我给药测定中的调节剂我获得该职业发展奖的目标是建立在我的治疗相关模型的功效之上。实验室的能力并为独立作为行为和分子研究的主要研究者做好准备我的实验室目前有能力进行拟议的研究，但我需要额外的分子药理学培训以了解 G 蛋白抗体捕获闪烁接近度我将在 Allyn Howlett 博士的实验室中学习完成目标 2 的分析方法，我还将学习补充知识。我还将受益于新英格兰生物实验室研讨会上的分子生物学技术。受体理论、变构模型的应用和信号偏差的量化，我将从我的我还需要接受我将学习的 Aim 3 实验的自我管理培训。在 Jenny Wiley 博士的实验室里，除了技术培训之外，我还需要职业发展以做好订单准备。为了研究独立性，我将接受资助、实验室管理、预算管理和负责任的研究进行这些职业发展活动将包括与共同导师的一对一培训以及研讨会，为我提供成功所需的培训。竞争 R01 资金并开发高效的独立研究项目。