In vitro and in vivo characterization of CB1 allosteric modulators

大麻素1(CB1)受体是治疗多种疾病的有效药物治疗靶点。 大量临床前和临床数据显示了治疗药物依赖/成瘾的有效性， 疼痛、肥胖/代谢综合征等。不幸的是，开发的药物也针对CB1受体 直接或间接取得的成功有限，即屈诺比诺产生不受欢迎的精神活动，利莫那班 会产生抑郁/自杀意念，而PF-04457845没有显示出疗效。重要考虑事项 在CB1激活的治疗和非治疗作用中，有多种信号通路起作用 这两件事中的任何一件。变构调节提供了一个额外的载体，CB1受体可以通过这个载体被 被操纵以获得治疗收益。这项研究建议1)检查已建立的CB1的新结构类似物 变构调节剂改变正构体配体结合、功能和信号偏向的能力；2) 正构和变构配体对N18TG2细胞G蛋白亚型依赖性偶联的影响 提供生理上相关的受体/G蛋白化学计量；以及3)评估CB1变构的效果 测定大麻素活性、阿片类药物戒断和自我给药的调节剂 在治疗相关模型中的疗效。我的这个职业发展奖的目标是在我的 实验室的能力，并为独立成为行为和分子领域的首席研究员做好准备 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 需要接受额外的分子药理学培训以了解G蛋白抗体捕获闪烁的近似性 完成目标2的化验，我将在Allyn Howlett博士的实验室学习。我也会学习互补的 来自新英格兰生物实验室的一个研讨会上的分子生物学技术。我也将从指导中受益 受体理论、变构模型的应用和信号偏倚的量化 共同导师特里·凯纳金博士。我还需要在Aim 3实验中接受自我管理方面的培训，我将学习这些内容 在珍妮·威利博士的实验室里。除了技术培训外，我还需要职业发展，以便为 为了研究的独立性。根据这一奖项，我将接受培训，包括实验技能、实验室管理、 预算管理，以及负责任的研究行为。这些职业发展活动将包括与共同导师的一对一培训，以及为我提供必要的培训以成功 争取R01资金，开发高生产力和高效率的独立研究计划。

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.