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.

我的研究的长期目标是调查调节细胞 - 细胞通信和神经传递的基本机制，重点是确定新颖的治疗靶标和治疗以预防和/或治疗药物滥用。我们的最初研究集中在阿片类药物和大麻素系统上，导致接收器相互作用复合物是新的治疗靶标，并鉴定了针对异源物的小分子。功绩奖使我能够将研究范围扩大到奖励途径中其他受体系统的范围，这导致发现了两个全新的神经肽受体系统。在延长期间，我打算在基本授予中描述的研究以及新表征的受体系统 - 这些内容如下所述。 将异二聚体选择配体的鉴定为疼痛治疗的新疗法：μor-Δor异构体配体：在基本赠款期间，我们使用高吞吐量筛选（HTS）来识别μor-Δor杂物的小分子激动剂。发现其中一种化合物具有与吗啡相当但耐受性和依赖性降低的镇痛特性（Gomes等，2013a）。该化合物的类似物是由MLPCN（加利福尼亚州拉霍拉）的化学家合成的。在绩效扩展期间，我们将使用体外和体内测定（镇痛，痛觉过敏，成瘾，奖励等）测试这些化合物。 HTS分析还产生了μor-Δor异构体的拮抗剂。筛选中心为我们提供了前90个命中。我们将使用体外测定法对它们进行表征，并测试小鼠（镇痛，痛觉过敏，异常性，成瘾，奖励等）中前5名候选者。 μor-Δor异构体选择性拮抗剂将是证明这些异源体在体内的物理意义的有价值工具。 CB1R-ΔOR配体：我们已经证明CB1R-ΔOR异构体是治疗神经性疼痛的靶标（Bushlin等，2012; Rozenfeld等，2012）和化学疗法诱导的疼痛（Sierra和Devi，在制备中）。我们最近开发了适合HTS的评估。在绩效延长期间，我计划进行HTS分析，以鉴定CB1R-ΔOR选择配体，并表征其挽救神经性疼痛的能力的最高点击。 确定奖励行为疾病的新治疗靶标： 神经肽在许多不同的物理过程中起着重要作用。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。这些肽中的一些参与调节进食，吸毒成瘾其他有益的行为。但是，这些肽激活的受体系统尚不清楚。在基本赠款期间，我们对两种prosaas衍生肽的受体除外（Gomes等，2013b; Gomes等人，综述），并使用受体的同源性模型在硅质筛选中鉴定出小分子（Wardman等人（Wardman等人），综述）。我们还发现这些受体与μor相互作用，但不相互作用，但这导致了新的药理学，这表明这些复合物可能是新型的治疗靶标。在绩效延长期间，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物的特性，识别和表征具有高亲和力和效能的小分子，并使用这些配体来探索这些受体在包括成瘾在内的奖励相关行为的调节中的作用。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打开了新的治疗可能性。 用新型脚手架来鉴定天然产物，以治疗疼痛和成瘾： 最近，我们确定了高度选择性的κOR配体（从肌肉C. maculata中提取）。我们发现，与其他KOR激动剂相比，Collybolide具有偏见的激动活性和更高的效力，可阻断非抗药性介导的瘙痒。由于其独特的脚手架，Collybolide可以接受广泛的多样化。在绩效延长期间，我计划与化学家合作，以产生类似物并以识别具有通缉效果（抗炎，抗吸毒，抗抑郁剂）和副作用降低的靶向κOR的意图。在先前的资金周期中，我们发现了一种新的机制（GPCR维度），并确定了新型的治疗靶标（最近去畸形的下丘脑受体和受体复合物）。在延长期间 治疗奖励行为的疾病，包括吸毒成瘾。