CB1 Allosteric Modulators: Molecular, Cellular and In Vivo Pharmacology

CB1 变构调节剂：分子、细胞和体内药理学

基本信息

批准号：
9056090
负责人：
DEBRA A KENDALL
金额：
$ 53.39万
依托单位：
UNIVERSITY OF CONNECTICUT STORRS
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-15 至 2021-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9056090
关键词：
2-arachidonylglycerol Active Sites Adverse effects Affinity Agonist Amides Animal Model Arrestins Attenuated Behavior Behavioral Model Binding Binding Sites Biochemistry Biological Biological Assay Biology Brain CNR1 gene CNR2 gene Cannabinoids Cannabis sativa plant Central Nervous System Diseases Complex Coupled Coupling Development Disease Dissociation Drug Addiction Drug abuse Eating Disorders Endocannabinoids Equilibrium Feeding behaviors G-Protein-Coupled Receptors GTP-Binding Proteins Goals Incidence Indoles Inflammation Knowledge Laboratories Lead Ligand Binding Ligands Mediating Memory Metabolic syndrome Molecular Molecular Profiling Mus Neuraxis Nicotine Outcome Outcome Study Pain Pathway interactions Pattern Pharmaceutical Chemistry Pharmacology Phosphorylation Physiological Processes Plants Process Property Regulation Research Rewards Signal Pathway Signal Transduction Site Structure-Activity Relationship Substance Addiction Substance abuse problem Synaptic Transmission System Testing Therapeutic Therapeutic Agents Translating Work anandamide base behavioral pharmacology cannabimimetics cannabinoid receptor design drug discrimination energy balance feeding functional group improved in vivo mRNA Expression mouse model novel painful neuropathy positive allosteric modulator public health relevance pyridine receptor response scaffold targeted treatment therapeutic target

项目摘要

DESCRIPTION (provided by applicant): The cannabinoid receptor CB1 is a G-protein coupled receptor (GPCR) that regulates neural transmission and other physiological processes. CB1 is activated by endogenous ligands (e.g. arachidonoyl ethanolamide (AEA) and 2-arachidonoyl glycerol (2-AG)) and various synthetic and plant-derived ligands that bind to the receptor orthosteric site. These CB1 orthosteric ligands transduce signals to downstream effectors primarily via Gi/o coupling, but Gs and arrestin coupling are also possible. The biological responses are implicated in many therapeutic applications, such as substance addiction, pain and inflammation, memory, and feeding and energy balance. Recently, allosteric modulators for GPCRs were discovered. Whereas positive allosteric modulators (PAMs) enhance the functional efficacy of orthosteric agonists, negative allosteric modulators (NAMs) non-competitively decrease the activity of the receptor. These modulators bind GPCR regions topographically distinct from the orthosteric ligand binding sites and thereby exert their modulatory effects in a highly subtype specific manner. CB1 NAMs offer exciting opportunities for developing therapeutics for drug addiction and metabolic syndromes and PAMs open a new pathway to treat pain-related conditions with a reduced incidence of unwanted psychotropic effects. However, the underlying mechanisms for CB1 allosteric modulation, including for allosteric-modulation biased downstream signaling, remain elusive. We have designed and synthesized highly potent PAMs, such as LDK 1256 (KB = 89 nM), and with high allostery, such as LDK1258 (=24.5), for enhanced agonist binding. We also have identified NAMs that are the only known modulators for reducing agonist binding to CB1. We have discovered that the CB1 allosteric modulator ORG27569 is coupled to an arrestin signaling pathway, which represents the first example of a CB1 biased allosteric modulator. In this project, we will synthesize compounds through optimizing the properties of identified allosteric modulators, develop new scaffolds and identify pharmacophoric groups that improve the equilibrium dissociation constants and cooperativity factors. These compounds will be tested for impact on agonist and inverse agonist binding. We will also elucidate their G-protein and arrestin coupling and downstream molecular-level activities. In an iterative process, key CB1 modulators will be fine tuned via structure-activity relationship (SAR) analysis. We will evaluate potent allosteric modulators for their impact on CB1 pharmacological responses in vivo. This effort involves assessing key CB1 allosteric modulators versus orthosteric compounds in functional assays including in the cannabinoid tetrad and drug discrimination assays. We will also test lead ligands in mouse models of neuropathic pain, feeding, and nicotine reward. The overall goal of this work is to identify new PAMs and NAMs, elucidate their molecular level profiles, and test lead ligands in animal models. These efforts are critical for elucidating the basis of CB1 allosteric modulation so that ultimately highly specific responses can be attained via therapeutic agents.

描述（申请人提供）：大麻素受体 CB1 是一种 G 蛋白偶联受体 (GPCR)，可调节神经传递和其他生理过程，CB1 被内源性配体（例如花生四烯酰乙醇酰胺 (AEA) 和 2-花生四烯酰甘油 (2) 激活）。 -AG)) 以及与受体正位点结合的各种合成配体和植物衍生配体。主要通过 Gi/o 耦合将信号转导至下游效应器，但 Gs 和抑制蛋白耦合也可能涉及许多治疗应用，例如物质成瘾、疼痛和炎症、记忆以及进食和能量平衡。已发现 GPCR 的变构调节剂（PAM）可增强正构激动剂的功能功效，而负变构调节剂（NAM）则非竞争性地降低 GPCR 的活性。这些区域调节剂与 GPCR 的结合与正位配体结合位点不同，从而以高度亚型特异性的方式发挥其调节作用，CB1 NAM 为开发药物成瘾和代谢综合征的治疗方法提供了令人兴奋的机会，并且 PAM 开辟了一条新的治疗途径。然而，我们设计的 CB1 变构调节的潜在机制（包括变构调节偏差心理下游信号传导）仍然难以捉摸。并合成了高效的 PAM，如 LDK 1256 (KB = 89 nM)，并具有高变构，如 LDK1258 (α=24.5)，用于增强激动剂结合。我们还发现 NAM 是唯一已知的减少激动剂的调节剂。我们发现 CB1 变构调节剂 ORG27569 与抑制蛋白信号通路偶联，这代表了 CB1 偏向变构调节剂的第一个例子。在这个项目中，我们将通过优化已确定的变构调节剂的特性来合成化合物，开发新的支架并确定可改善平衡解离常数和协同因子的药效基团。测试对激动剂和反向激动剂结合的影响，我们还将在迭代过程中阐明它们的 G 蛋白和抑制蛋白偶联以及下游分子水平的活性。我们将通过结构-活性关系 (SAR) 分析来评估有效的变构调节剂对 CB1 药理反应的影响。这项工作涉及在功能测定中评估关键的 CB1 变构调节剂与正构化合物，包括大麻素四联体。我们还将在神经性疼痛、进食和尼古丁奖励的小鼠模型中测试先导配体。这项工作的总体目标是识别新的 PAM 和尼古丁奖励。 NAM、阐明其分子水平概况并在动物模型中测试先导配体，这些努力对于阐明 CB1 变构调节的基础至关重要。以便最终可以通过治疗剂获得高度特异性的反应。