Post-translational regulation of opioid and cannabinoid receptors

阿片类药物和大麻素受体的翻译后调节

• 批准号: 9249714
• 负责人: Lakshmi A Devi
• 金额: $ 49.82万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9249714
• 关键词: Absence of pain sensation; Adverse effects; Affinity; Agonist; Analgesics; Antidepressive Agents; Award; Behavior; Biological Assay; CNR1 gene; Cannabinoids; Cell Communication; Cell physiology; Cells; Central Nervous System Diseases; Complex; Computer Simulation; Dependence; Dimerization; Disease; Drug Addiction; Drug abuse; Exhibits; Funding; G-Protein-Coupled Receptors; Goals; Grant; Homology Modeling; Hyperalgesia; Hypothalamic structure; In Vitro; Ligands; Mediating; Morphine; Mus; Names; Natural Products; Neuropeptide Receptor; Neuropeptides; Normal Cell; Opioid; Opioid Receptor; Pain; Pain management; Pathway interactions; Peptides; Pharmacology; Physiological; Physiological Processes; Play; Post-Translational Regulation; Preparation; Prevention; Property; Proteins; Pruritus; Regulation; Research; Rewards; Role; Signal Transduction; System; Technology; Testing; Therapeutic; Work; addiction; allodynia; analog; base; cannabinoid receptor; chemotherapy; dimer; feeding; high throughput screening; in vitro Assay; in vivo; neurotransmission; new therapeutic target; novel; novel therapeutics; optogenetics; painful neuropathy; receptor; scaffold; screening; small molecule; targeted treatment; tool

The long-term goal of my research has been to investigate the fundamental mechanisms modulating cell-cell communication and neurotransmission with a focus on identifying novel therapeutic targets and therapeutics for the prevention and/or treatment of drug abuse. Our initial studies, which were focused on the opioid and cannabinoid systems, led to the identification of receptor interacting complexes as novel therapeutic targets as well as identification of small molecules targeting heteromers. The MERIT award has enabled me to broaden the scope of research to other receptor systems in the reward pathway and this led to the discovery of two completely new neuropeptide receptor systems. During the extension period I intend to pursue studies described in the base grant as well as the newly characterized receptor systems – these are described below. Identification of heterodimer-selective ligands as new therapeutics for the treatment of pain: μOR-δOR heteromer ligands: During the base grant period, we used high throughput screening (HTS) to identify small molecule agonists of the μOR-δOR heteromer. One of the compounds was found to have analgesic properties comparable to morphine but with reduced tolerance and dependence (Gomes et al., 2013a). Analogs of this compound are being synthesized by chemists at MLPCN (LaJolla, CA). During the MERIT extension period we will test these compounds using in vitro and in vivo assays (analgesia, hyperalgesia, addiction, reward etc). The HTS analysis has also yielded antagonists of the μOR-δOR heteromer. The screening center has provided us with top 90 hits. We will characterize them using in vitro assays and test the top 5 candidates in mice (analgesia, hyperalgesia, allodynia, addiction, reward etc). A μOR-δOR heteromer selective antagonist will be a valuable tool for the demonstration of the physiological significance of these heteromers in vivo. CB1R-δOR ligands: We have demonstrated CB1R-δOR heteromers to be a target for the treatment of neuropathic pain (Bushlin et al., 2012; Rozenfeld et al., 2012) and chemotherapy-induced pain (Sierra and Devi, in preparation). We have recently developed an assay that is suitable for HTS. During the MERIT extension period, I plan to carry out HTS analysis for the identification of CB1R-δOR-selective ligands and characterize the top hits for their ability to relieve neuropathic pain. Identification of new therapeutic targets for disorders of reward behaviors: Neuropeptides play important roles in a number of diverse physiological processes. Some of the most abundant neuropeptides found in the hypothalamus are generated from the protein named proSAAS; some of these peptides are involved in modulating feeding, drug addiction other rewarding behaviors. However, the receptor systems activated by these peptides were not known. During the base grant period, we deorphanized receptors for two of the proSAAS-derived peptides (Gomes et al., 2013b; Gomes et al., under review) and identified small molecules by in silico screening using a homology model of the receptor (Wardman et al., under review). We also found that these receptors interact with μOR but not δOR and this leads to novel pharmacology suggesting that these complexes could be novel therapeutic targets. During the MERIT extension period, I intend to characterize the properties of these receptors complexes using state-of-the-art technologies (Optogenetics and DREADD), identify & characterize small molecules with high affinity & potency, and use these ligands to explore a role for these receptors in the regulation of reward-related behaviors including addiction. These studies are likely to have a major impact on the field since they open novel therapeutic possibilities for the treatment of a variety of CNS disorders including drug addiction. Identification of natural products with novel scaffolds for the treatment of pain and addiction: Recently, we identified Collybolide (extracted from the mushroom C. maculata), as a highly selective κOR ligand. We find that Collybolide exhibits biased agonistic activity and a 10-fold higher potency for blocking non-histamine-mediated itch as compared to other KOR agonists. Due to its unique scaffold, Collybolide is amenable to extensive diversification. During the MERIT extension period I plan to work with chemists to generate analogs and characterize them with the intent of identifying compounds targeting κOR with wanted effects (antipruritis, anti-addiction, antidepressant) and reduced side-effects. During the previous funding cycles we have discovered a novel mechanism (GPCR dimerization) and have identified novel therapeutic targets (recently deorphanized hypothalamic receptors and receptor complexes). During the extension period we intend to demonstrate the relevance of these new systems to normal cell function and to disease states using state-of-the art technologies and identify therapeutics for the treatment of disorders of the reward behaviors including drug addiction.

我这项研究的长期目标是研究调节细胞间通讯和神经传递的基本机制，重点是确定预防和/或治疗药物滥用的新治疗靶点和治疗方法。我们最初的研究集中在阿片和大麻素系统上，导致将受体相互作用的复合体确定为新的治疗靶点，并确定了针对异构体的小分子。优秀奖使我能够将研究范围扩大到奖励途径中的其他受体系统，这导致了两个全新的神经肽受体系统的发现。在延期期间，我打算继续进行基本赠款中描述的研究以及新表征的受体系统--这些描述如下。 异二聚体选择性配体作为治疗疼痛的新疗法的鉴定：μOR-δ或异构体配体：在基础拨款期间，我们使用高通量筛选(HTS)来鉴定μOR-δOR异构体的小分子激动剂。其中一种化合物被发现具有与吗啡相当的止痛特性，但耐受性和依赖性更低(戈麦斯等人，2013a)。MLPCN(加利福尼亚州拉霍亚)的化学家正在合成这种化合物的类似物。在疗效延长期内，我们将使用体外和体内测试(止痛、痛觉过敏、成瘾、奖励等)来测试这些化合物。高温超导分析还发现了μ或-δ或异构体的拮抗剂。放映中心为我们提供了前90名的点击率。我们将使用体外试验对它们进行表征，并在小鼠身上测试排名前5位的候选药物(止痛、痛觉过敏、异常痛觉、成瘾、奖励等)。μ或-δ或异构体选择性拮抗剂将是在体内展示这些异构体的生理意义的有价值的工具。 CB1R-δ或配体：我们已经证明CB1R-δ或异构体是治疗神经性疼痛(Bushlin等人，2012年；Rozenfeld等人，2012年)和化疗引起的疼痛(Sierra和Devi，正在准备中)的靶点。我们最近开发了一种适用于HTS的检测方法。在优点延长期间，我计划进行高温超导分析，以确定CB1R-δ或-选择性配体，并表征最热门的HITS具有缓解神经病理性疼痛的能力。 奖赏行为障碍新治疗靶点的确定 神经肽在许多不同的生理过程中发挥着重要作用。在下丘脑中发现的一些最丰富的神经肽是由一种名为proSAAS的蛋白质产生的；其中一些肽参与调节摄食、药物成瘾和其他奖励行为。然而，这些多肽激活的受体系统尚不清楚。在基础赠款期间，我们使两个proSAAS衍生多肽的受体去孤儿(Gome等人，2013b；Gome等人，正在审查中)，并通过使用受体的同源模型进行电子筛查来鉴定小分子(Wardman等人，正在审查中)。我们还发现，这些受体与μOR相互作用，但不与δOR相互作用，这导致了新的药理作用，表明这些络合物可能是新的治疗靶点。在优点延长期间，我打算使用最先进的技术(光遗传学和DREADD)来表征这些受体复合体的性质，识别和表征具有高亲和力和效力的小分子，并使用这些配体来探索这些受体在调节包括成瘾在内的奖赏相关行为中的作用。这些研究可能会对该领域产生重大影响，因为它们为治疗包括药物成瘾在内的各种中枢神经系统疾病开辟了新的治疗可能性。 鉴定具有治疗疼痛和成瘾的新型支架的天然产物： 最近，我们确定Collybolide(从蘑菇C.maculata中提取)是一种高度选择性的κ或配体。我们发现，与其他KOR激动剂相比，Collybolide表现出偏向的激动剂活性，并且阻断非组胺介导的瘙痒的效力是其他KOR激动剂的10倍。由于其独特的支架，Collybolide可以进行广泛的多样化。在优点延期期间，我计划与化学家合作生成类似物并对其进行表征，目的是识别针对κ或具有想要的效果(止痒、抗成瘾、抗抑郁)并减少副作用的化合物。在之前的资助周期中，我们发现了一种新的机制(GPCR二聚体)，并确定了新的治疗靶点(最近去吗啡的下丘脑受体和受体复合体)。在延长期间，我们打算使用最先进的技术证明这些新系统与正常细胞功能和疾病状态的相关性，并确定治疗 治疗包括药物成瘾在内的奖赏行为障碍。