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疗法。我们最初的研究重点是阿片类药物和大麻素系统，最终确定了受体相互作用复合物作为新的治疗靶点，并确定了靶向异聚体的小分子。优异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。其中一种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，2013a）。 MLPCN（加利福尼亚州拉霍亚）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测定（镇痛、痛觉过敏、成瘾、奖励等）来测试这些化合物。 HTS 分析还产生了 μOR-δOR 异聚体的拮抗剂。放映中心为我们提供了前 90 部热门影片。我们将使用体外测定来表征它们，并在小鼠中测试前 5 种候选药物（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用于 HTS 的检测方法。在 MERIT 延长期间，我计划进行 HTS 分析来鉴定 CB1R-δOR 选择性配体，并表征最热门的配体缓解神经性疼痛的能力。 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。然而，这些肽激活的受体系统尚不清楚。在基础拨款期间，我们对两种 proSAAS 衍生肽的受体进行去孤儿化（Gomes 等人，2013b；Gomes 等人，正在审查中），并使用受体的同源模型通过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，这产生了新的药理学，表明这些复合物可能是新的治疗靶点。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和效力的小分子，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，与其他 KOR 激动剂相比，Collybolide 表现出偏向的激动活性，且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，目的是识别具有所需效果（止痒、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。