New 5HT2AR Target Identification and Assay Development for Discovering Psychoplastogenic Compounds

用于发现精神质体化合物的新 5HT2AR 靶标鉴定和检测开发

• 批准号: 10742536
• 负责人: Haifeng Eishingdrelo
• 金额: $ 31.93万
• 依托单位: BIOINVENU CORPORATION
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10742536
• 关键词: Acceleration; Affinity; Aftercare; Agonist; Behavior; Binding; Biological Assay; Brain; Cell physiology; Cerebral cortex; Chromosome Pairing; Clinical; Complex; Dangerousness; Data; Dendritic Spines; Drug Addiction; Drug Screening; Drug Use Disorder; Endocytosis; Enzymes; Experimental Designs; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; HTR2A gene; Hallucinogens; Health Professional; Ketanserin; Libraries; Ligands; Link; Location; Mediating; Mental Depression; Mental Health; Mental disorders; Molecular; Mus; Neurologic; Neuronal Differentiation; Neuronal Plasticity; Neurons; Opioid Receptor; Pathway interactions; Persons; Phase; Phosphotransferases; Physiological; Play; Protein Isoforms; Protein Kinase; Proteins; Receptor Signaling; Research; Rodent; Role; Scientist; Screening Result; Serotonin Antagonists; Signal Pathway; Signal Transduction; Signaling Protein; Technology; Therapeutic; Therapeutic Effect; Therapeutic Equivalency; Toxic effect; antagonist; assay development; behavior test; beta-arrestin; drug action; drug development; drug discovery; experience; flexibility; improved; in vivo; insight; interest; neurite growth; neuroimaging; neuropsychiatric disorder; new therapeutic target; novel therapeutics; opioid use; pharmacologic; receptor; receptor mediated endocytosis; serotonin receptor; side effect; synaptogenesis; therapeutic development; trafficking; transcription factor

Specific Aim: New 5HT2AR/14-3-3 isoform signal pathway identification and assay development for discovering psychoplastogenic compounds. Significance: Emerging data and limited clinical results show the promise of psychedelic drugs in the durable relief of people from seemingly untreatable psychiatric disorders and drug addictions. Neuroimaging evidence supports the concept that the brain after a psychedelic treatment becomes more flexible or `plastic', which is thought to be a component of its putative therapeutic effects. However, the side effects of psychedelics can be dangerous or even deadly. To develop new therapeutic options, we need to understand psychedelics underlying mechanisms of action. Many psychedelics act on the 5-HT2A serotonin receptor (5HT2AR), a G-protein Coupled Receptor (GPCR), to exert their effects. G-protein and β-arrestin signaling pathways cannot distinguish therapeutic and side effects of psychedelics. Importantly, the 5HT2AR antagonist ketanserin, can promote receptor endocytosis and still derive equivalent relief from depression-associated behaviors in mice, suggesting trafficking plays an important role. We studied GPCR/14-3-3 signal adaptor interactions using the LinkLight technology and found that GPCR/14-3-3 signals are closely related to receptor trafficking. Thus, studying 5HT2AR/14-3-3 signals mediated by psychedelics should shed light on understanding molecular mechanisms that contribute to psychedelic and therapeutic benefits, and develop new psychoplastogenic compounds. We have developed 5HT2AR/14-3-3ε and 5HT2AR/14-3-3γ LinkLight assays. We plan to develop additional 5HT2AR/14-3-3 isoform assays including 14-3-3β and 14-3-3ζ since a GPCR can differentially and temporally engage with 14-3-3 isoforms. Different 14-3-3 isoforms could display different functions. We plan to profile a panel of psychedelics in 5HT2AR/14-3-3 isoform assays and see if we can connect signaling pathways to physiological functions. In addition, we want to conduct a pilot screen to find compounds biased to the 14-3-3 signaling pathway without G-protein signals. Such biased compounds could lead us to better understand psychedelics' underlying mechanisms of action and develop new psychoplastogenic compounds. Experimental Design: Task 1. Develop additional 5HT2AR/14-3-3β and 5HT2AR/14-3-3ζ assays, profile 5HT2AR/14-3-3 isoform (ε, γ, β, & ζ) pathways with a panel of psychedelics including hallucinogenic and non- hallucinogenic agonists, as well as antagonists, and differentially and temporally characterize 5HT2AR/14-3-3 isoform signals. Task 2. Identify pathway-biased ligands by conducting a pilot screen with an in-house GPCR-focused library and characterize hits activities in 5HT2AR/14-3-3 isoforms, G-protein and β-arrestin pathway assays. Next Phase Plan: We expect to find multi-pathway biased ligands based on our results of screens using opioid receptor/14- 3-3 assays (unpublished results). These multi-pathway biased ligands will be evaluated for preliminary toxicity and DMPK activities and for in vivo rodent behavior tests and neuroimaging. The proof of concept studies will enable us to partner with potential collaborators to accelerate drug development.

具体目的： 新的5HT2AR/14-3-3同工型信号途径识别和测定开发用于发现 精神构成化合物。 意义： 新兴数据和有限的临床结果表明，迷幻药物在持久的缓解中的希望 看似无法治疗的精神疾病和吸毒的人。神经影像证据 支持以下概念，即迷幻治疗后的大脑变得更加灵活或“塑料” 被认为是其假定治疗作用的组成部分。但是，迷幻药的副作用 可能是危险的，甚至是致命的。要开发新的治疗选择，我们需要了解 迷幻动作机制的潜在机制。 关于5-HT2A 5-羟色胺受体（5HT2AR）的许多迷幻作用，G蛋白偶联受体 （GPCR），执行其效果。 G蛋白和β-arrestin信号传导途径无法区分治疗 和迷幻的副作用。重要的是，5HT2AR拮抗剂酮酮可以促进接收器 内吞作用，仍然可以从小鼠中抑郁相关行为的同等缓解，这表明 贩运起着重要作用。 我们使用Linklight技术研究了GPCR/14-3-3信号适配器相互作用，发现 GPCR/14-3-3信号与受体贩运密切相关。这是研究5HT2AR/14-3-3信号 通过迷幻而介导的，应阐明理解有助于的分子机制 迷幻和治疗的益处，并开发新的精神构成化合物。 我们已经开发了5HT2AR/14-3-3ε和5HT2AR/14-3-3γ链接测定。我们计划发展 由于GPCR可以差异化 并暂时与14-3-3同工型互动。不同的14-3-3同工型可以显示不同的功能。 我们计划在5HT2AR/14-3-3同工型分析中介绍一个迷幻小组，看看我们是否可以连接 信号通路通往物理功能。此外，我们想进行一个飞行员屏幕以查找 化合物偏向没有G蛋白信号的14-3-3信号通路。这种偏见的化合物 可能会导致我们更好地了解迷幻的基本行动机制并发展新的 精神构成化合物。 实验设计： 任务1。开发额外的5HT2AR/14-3-3β和5HT2AR/14-3-3ζ分析，配置文件5HT2AR/14-3-3 同工型（ε，γ，β和ζ）途径，具有一系列迷幻药，包括致幻和非 - 致幻激动剂以及拮抗剂，以及差异化和暂时的特征 5HT2AR/14-3-3同工型信号。 任务2。通过以内部GPCR为中心进行飞行员屏幕来识别途径偏见的配体 图书馆并表征5HT2AR/14-3-3同工型，G蛋白和β-arrest素途径中的命中活动 测定。 下一个阶段计划： 我们希望使用阿片类药物接收器/14-基于我们的筛选结果找到多条道路的偏置配体。 3-3分（未发表的结果）。这些多条纹偏见的配体将进行初步评估 毒性和DMPK活性以及体内啮齿动物行为测试和神经影像学。概念证明 研究将使我们能够与潜在的合作者合作，以加速药物开发。