An optimized screening platform for identifying and quantifying biased agonists as drugs for the treatment of Opioid Use Disorder

用于识别和量化偏向激动剂作为阿片类药物使用障碍治疗药物的优化筛选平台

• 批准号: 10334560
• 负责人: THOMAS E HUGHES
• 金额: $ 31.54万
• 依托单位: MONTANA MOLECULAR, LLC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10334560
• 关键词: Adverse effects; Agonist; American; Americas; Animal Model; Biological; Biological Assay; Biosensor; CNR1 gene; Cannabinoids; Cells; Certification; Chinese Hamster Ovary Cell; Clinical Research; Computer software; Cyclic AMP; Data; Data Analyses; Dependence; Development; Devices; Dopamine; Dose; Drug Targeting; Fluorescence; G protein coupled receptor kinase; GTP-Binding Proteins; Goals; Helping to End Addiction Long-term; Individual; Industry; Kinetics; Ligands; Manuals; Measurement; Measures; Methods; Molecular; Monitor; National Institute of Drug Abuse; ORL1 receptor; Opioid; Opioid Receptor; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phase; Phosphotransferases; Production; Protocols documentation; Reader; Reagent; Receptor Signaling; Reproducibility; Research; Research Personnel; Running; Signal Pathway; Signal Transduction; Site; Site Visit; Speed; Structure-Activity Relationship; System; Testing; Therapeutic; Time; United States; Videoconferencing; Viral Vector; analytical tool; base; beta-arrestin; drug action; drug discovery; field study; improved; in vivo; manufacturing scale-up; new therapeutic target; non-opioid analgesic; novel therapeutics; opioid epidemic; opioid misuse; opioid use disorder; quality assurance; receptor; response; screening; tool; validation studies

Millions of Americans today have an opioid use disorder (OUD). Millions more misuse opioids, and the crisis continues to grow. The goal of this proposal is to speed the discovery of non-addictive analgesics by providing drug discovery teams with simpler, more robust, more quantitative, assays for agonist bias. Driven by the urgency of the problem we are seeking Fast Track support to create new assay and analytic tools for drug discovery in OUD research. Our goal is to optimize and test new assays for agonist bias at particular receptors that couple to both the Gi and β-arrestin signaling pathway, and create new tools to improve the analysis of structure/activity relationships. There are good reasons to search for biased agonists to the receptors identified in the NIDA “top ten” list of medication development priorities. Biased agonists could activate beneficial signaling pathways while avoiding those that cause adverse effects. Finding these biased agonists is difficult: current assays for detecting bias, while established and validated, suffer from drawbacks that are limiting translatability to animal models and clinical studies. These include entirely different sets of experimental conditions for measuring the different signaling pathways being compared and different time courses of the response being measured. The latter results in time-dependence of the bias measurement which complicates predictions of in vivo efficacy and complicates SAR tables by adding extra variables. Our new assay will simultaneously measure the kinetics of Gi and β-arrestin signaling in living cells. This project will involved creating new tools as well as re-purposing ones we have already developed to study non-OUD drug targets. The assay will be optimized for use on standard fluorescence plate readers, and a data analysis toolbox will be developed to simplify quantification of agonist bias based on kinetic measurements. Phase I will complete the initial validation studies on the NOP opioid receptor, with goal of demonstrating assay reliability and sensitivity milestones. Phase II will optimize the assay for D3 dopamine, CB1 cannabinoid and OPRM1 opioid receptors and develop the analysis toolbox for deployment on standard plate readers and software packages commonly used in drug discovery. In the second half of Phase II, assays with detailed protocols will be ready distribute to researchers who are developing new drugs for OUD.

如今，数百万的美国人患有绿化疾病（OUD）。数以百万计的小姐 Opioids，危机继续增长。该提议的目的是加快 通过提供更简单的药物发现团队来发现非添加性镇痛药 对激动剂偏见的更强大，更定量的测定法。受到紧迫的驱动 我们正在寻找快速轨道支持的问题，以创建新的测定和分析工具 OUD研究中的药物发现。我们的目标是优化和测试新测定 在gi和β-arrestin信号传导的特定受体上的激动剂偏见 途径，并创建新工具来改善结构/活动关系的分析。 有充分的理由将偏见的激动剂搜索到所确定的接收器 NIDA“十大”药物开发优先事项清单。有偏的激动剂可以激活 有益的信号通路，同时避免造成不良影响的信号通路。发现 这些偏见的激动剂很困难：当前检测偏见的测定法，同时建立 经过验证，遭受限制转化性为动物模型的缺点 和临床研究。这些包括完全不同的实验条件集 测量要比较不同的信号通路和不同的时间课程 响应正在测量。后来导致偏差的时间依赖性 测量使体内效率的预测复杂化并使SAR复杂化 通过添加额外的变量来表。 我们的新测定将简单地测量GI和β-arrestin信号传导的动力学 活细胞。该项目将涉及创建新工具以及重新设计工具 我们已经开发用于研究非饮用药物靶标。该测定将优化 用于标准荧光板读取器，数据分析工具箱将是 开发以简化基于动力学测量值的激动剂偏差的数量。 第一阶段将完成有关NOP阿片类药物受体的初始验证研究，目标 证明测定的可靠性和敏感性里程碑。第二阶段将优化 D3多巴胺，CB1大麻素和OPRM1阿片类药物受体的测定，并发展 用于在标准板读取器和软件包上部署的分析工具箱 通常用于药物发现。在第二阶段的后半部分，用详细的测定 协议将准备向正在为OUD开发新药的研究人员分发。