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。我们最初的研究主要集中在阿片类药物和大麻素系统，导致识别受体相互作用复合物作为新的治疗靶点，以及识别靶向异聚体的小分子。MERIT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。在延长期内，我打算继续进行基本资助中描述的研究以及新表征的受体系统-这些在下文中描述。 识别异二聚体选择性配体作为治疗疼痛的新疗法：μOR-δOR异聚体配体：在基本资助期间，我们使用高通量筛选（HTS）来识别μOR-δOR异聚体的小分子激动剂。发现其中一种化合物具有与吗啡相当的镇痛性质，但耐受性和依赖性降低（Gomes等人，2013年a）。MLPCN（LaJolla，CA）的化学家正在合成该化合物的类似物。在MERIT延长期内，我们将使用体外和体内试验（镇痛、痛觉过敏、成瘾、奖励等）检测这些化合物。HTS分析还产生了μOR-δOR异聚体的拮抗剂。放映中心给我们提供了90首热门歌曲。我们将使用体外测定来表征它们，并在小鼠中测试前5个候选物（镇痛、痛觉过敏、异常性疼痛、成瘾、奖励等）。μOR-δOR异聚体选择性拮抗剂将是证明这些异聚体在体内的生理意义的有价值的工具。 CB 1 R-δOR配体：我们已经证明CB 1 R-δOR异聚体是治疗神经性疼痛的靶标（Bushlin et al.，2012; Rozenfeld等人，2012）和化疗引起的疼痛（Sierra和Devi，准备中）。我们最近开发了一种适用于HTS的检测方法。在MERIT延长期内，我计划进行HTS分析，以鉴定CB 1 R-δ OR选择性配体，并表征其缓解神经性疼痛的能力。 识别奖励行为障碍的新治疗靶点： 神经肽在许多不同的生理过程中发挥重要作用。在下丘脑中发现的一些最丰富的神经肽是由名为proSAAS的蛋白质产生的;其中一些肽参与调节进食，药物成瘾和其他奖励行为。然而，这些肽激活的受体系统是未知的。在基础资助期间，我们对两种proSAAS衍生肽的受体进行去乙酰化（Gomes等人，2013 b; Gomes等人，正在审查中）并通过使用受体的同源模型的计算机筛选鉴定小分子（Wardman等，审查中）。我们还发现这些受体与μOR而不是δOR相互作用，这导致了新的药理学，表明这些复合物可能是新的治疗靶点。在MERIT扩展期间，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物的特性，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奖励相关行为（包括成瘾）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CNS疾病开辟了新的治疗可能性。 用于治疗疼痛和成瘾的具有新型支架的天然产物的鉴定： 最近，我们确定了Collybolide（从蘑菇C。maculata），作为高度选择性的κOR配体。我们发现，Collybolide表现出偏向激动活性和10倍以上的效力，阻断非组胺介导的瘙痒相比，其他KOR激动剂。由于其独特的支架，Collybolide适合广泛的多样化。在MERIT延长期内，我计划与化学家合作生成类似物并对其进行表征，目的是鉴定靶向κOR的化合物，这些化合物具有预期的作用（抗癫痫、抗成瘾、抗抑郁）和减少的副作用。在之前的资助周期中，我们发现了一种新的机制（GPCR二聚化），并确定了新的治疗靶点（最近去乙酰化的下丘脑受体和受体复合物）。在延长期内，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，并确定治疗方法。 包括药物成瘾在内的奖赏行为障碍的治疗。