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

基本信息

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
项目类别：
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

项目摘要

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疗法。关于阿片类药物和大麻素系统相互作用的研究，导致识别出受体复合物作为新的治疗靶点，以及识别出针对异聚体的小分子。 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。这导致了两个发现全新的神经肽受体系统。在延长期内，我打算继续进行基础拨款中描述的研究以及新表征的受体系统——这些内容如下所述。鉴定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：在基础资助期间，我们使用高通量筛选（HTS）来鉴定μOR-δOR异聚体的小分子激动剂之一。化合物被发现具有与吗啡相当的镇痛特性，但耐受性和依赖性降低（Gomes 等人，2013a）。在 MERIT 延长期内，我们将使用体外和体内测定（镇痛、痛觉过敏、成瘾、奖励等）来测试这些化合物。筛选中心为我们提供了前 90 个候选药物，我们将使用体外测定来表征它们，并在小鼠中测试前 5 个候选药物（镇痛、痛觉过敏、 μOR-δOR异聚体选择性拮抗剂将成为证明这些异聚体在体内的生理意义的有价值的工具。 CB1R-δOR 配体：我们已经证明 CB1R-δOR 异聚体是治疗神经性疼痛（Bushlin 等人，2012；Rozenfeld 等人，2012）和化疗引起的疼痛（Sierra 和 Devi，准备中）的靶点我们最近开发了一种适合 HTS 的检测方法，在 MERIT 延长期内，我计划进行 HTS 分析来识别。 CB1R-δOR-选择性配体，并描述了其缓解神经性疼痛能力的热门产品。确定奖赏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；其中一些肽参与调节进食、药物成瘾和其他奖励行为。在基础授予期间，我们对两种 proSAAS 衍生肽的受体进行去孤儿化处理（Gomes 等人，2013b；Gomes 等人）。等人，正在审查中）并通过使用受体同源模型进行计算机筛选来鉴定小分子（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，这导致了新的药理学提示。在 MERIT 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的小分子。和效力，并使用这些配体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，因为它们为治疗各种疾病开辟了新的治疗可能性。中枢神经系统疾病，包括毒瘾。鉴定具有治疗疼痛和成瘾的新型支架的天然产物：最近，我们发现 Collybolide（从蘑菇 C. maculata 中提取）是一种高度选择性的 κOR 配体，我们发现 Collybolide 表现出偏向的激动活性，与其他 KOR 相比，其阻断非组胺介导的瘙痒的效力高出 10 倍。由于其独特的支架，Collybolide 适合广泛的多样化，在 MERIT 延长期间，我计划与化学家合作生产类似物和药物。对它们进行表征，目的是识别具有所需效果（抗瘙痒、抗成瘾、抗抑郁）并减少副作用的靶向 κOR 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 二聚化）并确定了新的治疗靶点（最近脱孤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。治疗奖赏行为障碍，包括毒瘾。