In vitro and in vivo characterization of CB1 allosteric modulators

大麻素1型（CB 1）受体是许多疾病的有希望的药物靶标， 大量临床前和临床数据证明了治疗药物依赖/成瘾的功效， 疼痛、肥胖/代谢综合征和其他。不幸的是，针对CB 1受体开发的药物 直接或间接的成功有限，即屈大麻酚产生不希望的精神活性，利莫那班 产生抑郁/自杀意念，PF-04457845未显示疗效。重要的考虑因素 在CB 1激活的治疗与非治疗作用中， 这两个人中的任何一个变构调节提供了一种额外的载体，通过该载体可以将CB 1受体 为了获得治疗效果本研究提出1）检查已建立的CB 1的新结构类似物 变构调节剂，其改变正构配体结合、功能和信号传导偏好的能力; 2） 在N18 TG 2细胞中通过正构和变构配体表征G蛋白亚型依赖性偶联， 提供生理学相关的受体/G蛋白化学计量;和3）评估CB 1变构的作用 调节剂在测定大麻素活性、阿片样物质戒断和自我给药中的应用 在治疗相关模型中的疗效。我的目标是这个职业发展奖是建立在我的 实验室的能力，并准备作为行为和分子生物学的主要研究者独立 药物滥用的药理学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，以学习G蛋白抗体捕获闪烁接近 我将在Allyn Howlett博士的实验室学习如何完成目标2。我也会学习互补 在新英格兰生物实验室的一个研讨会上，我也将受益于指导， 受体理论，别构模型的应用和信号偏差的量化，我将从我的 共同导师特里·肯纳金博士我还需要接受Aim 3实验的自我管理培训 在珍妮·威利博士的实验室里除了技术培训，我还需要职业发展， 研究独立性。在这个奖项下，我将接受培训，在granitary，实验室管理， 预算管理和负责任的研究行为。这些职业发展活动将包括与共同导师的一对一培训，以及为我提供必要培训的研讨会， 竞争R 01资金，并开发一个高产高效的独立研究计划。

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.