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: 9911512
• 负责人: THOMAS E HUGHES
• 金额: $ 22.11万
• 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 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9911512
• 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; Drug effect disorder; Fluorescence; G protein coupled receptor kinase; GTP-Binding Proteins; Goals; 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; Viral Vector; analytical tool; base; beta-arrestin; 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; prescription opioid; quality assurance; receptor; response; screening; symposium; tool; validation studies

! Project Summary 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.

！ 项目概要 如今，数以百万计的美国人患有阿片类药物使用障碍 (OUD)，还有数百万人滥用阿片类药物。 阿片类药物，危机继续加剧 该提案的目标是加快阿片类药物的使用。 通过为药物发现团队提供更简单的方法来发现非成瘾性镇痛药 在紧迫性的推动下，对激动剂偏差进行更稳健、更定量的分析。 我们正在寻求快速通道支持来创建新的检测和分析工具 OUD 研究中的药物发现我们的目标是优化和测试新的检测方法。 与 Gi 和 β-arrestin 信号传导耦合的特定受体的激动剂偏倚 路径，并创建新工具来改进结构/活动关系的分析。 有充分的理由去寻找针对在 NIDA 药物开发优先事项“十大”清单可能会激活偏向激动剂。 有益的信号通路，同时避免那些造成不利影响的信号通路。 这些有偏差的激动剂很困难：目前检测偏差的测定方法，虽然已建立 并经过验证，但遇到了限制转化为动物模型的问题 和临床研究。这些包括完全不同的实验条件。 测量正在比较的不同信号通路和不同的时间进程 后者导致偏差的时间依赖性。 测量使体内功效的预测变得复杂，并使 SAR 变得复杂 通过添加额外变量的表。 我们的新检测方法将同时测量 Gi 和 β-arrestin 信号传导的动力学 该项目将涉及创建新工具以及重新利用这些工具。 我们已经开发了用于研究非 OUD 药物靶点的检测方法，并将对其进行优化。 用于标准荧光板读取器，并且将提供数据分析工具箱 开发用于简化基于动力学测量的激动剂偏差的量化。 第一阶段将完成 NOP 阿片受体的初步验证研究，目标 证明检测可靠性和灵敏度里程碑的第二阶段将优化 检测 D3 多巴胺、CB1 大麻素和 OPRM1 阿片受体并开发 用于部署在标准酶标仪和软件包上的分析工具箱 常用于药物发现，在第二阶段的后半部分进行详细的分析。 协议将准备好分发给正在开发 OUD 新药的研究人员。