Development of G protein beta gamma subunit inhibitors to improve the safety and efficacy of opioid analgesics

开发G蛋白βγ亚基抑制剂以提高阿片类镇痛药的安全性和有效性

基本信息

批准号：
10026089
负责人：
EMILY M JUTKIEWICZ
金额：
$ 70.83万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-30 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10026089
关键词：
ADRBK1 gene Absence of pain sensation Acute Acute Pain Address American Analgesics Animal Disease Models Animals Arrestins Binding Brain Characteristics Chemicals Chemistry Chemosensitization Chronic Chronic inflammatory pain Constipation Coupled Data Development Dyes Electrophysiology (science)Family Feedback G protein coupled receptor kinase G-protein Beta gamma GTP-Binding Protein alpha Subunits, Gs GTP-Binding Proteins Goals Gold Knock-out Knockout Mice Laboratories Lead Mediating Modeling Morphine Mus Opioid Opioid Analgesics Opioid Receptor Opioid agonist Pain Pathway interactions Patients Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Phospholipase C Phosphorylation Phosphotransferases Physical Dependence Potassium Channel Process Property Protein Inhibition Protein Subunits Proteins Publishing Rattus Receptor Signaling Regulation Reporting Rewards Role Safety Scheme Self Administration Series Signal Transduction Slice Synaptic Membranes Testing Therapeutic Time Ventilatory Depression Vesicle addiction arrestin 2 base chronic pain cost gallein high throughput screening improved in vitro testing in vivo inhibitor/antagonist lead candidate mouse model mu receptors neurotransmitter release novel novel drug class novel strategies novel therapeutics opioid use phospholipase C gamma prototype receptor scaffold side effect small molecule inhibitor standard care therapeutic development

项目摘要

This application proposes a novel approach to improving the safety of opioid analgesics by post-receptor pharmacological targeting of G protein  subunits to modify the actions of -opioid receptors (MORs). Our laboratory has identified small molecule inhibitors of G protein  subunits that demonstrate in vivo efficacy in various animal models of disease. Relevant to this application, we identified two related G inhibitors, M119 and gallein that increase MOR agonist analgesic potency in mice without potentiating side effects that include development of tolerance, respiratory depression, constipation or addiction. Thus co-administration of G inhibitors with opioid analgesics has the potential to improve their safety profile by opening up the therapeutic window between analgesic efficacy and deleterious side effects. We propose that gallein and M119 modify opioid action by blocking specific feedback pathways downstream of MORs while leaving pro-analgesic pathways intact. MORs couple to the Gi family of G proteins and promote analgesia through G protein-dependent inhibition of neurotransmitter release via G-dependent regulation of K+ (GIRK) channels, N-type Ca2+ channels and inhibition of vesicle fusion with synaptic membranes. At the same time G activates feedback pathways including phospholipase C (PLC) and G protein-coupled receptor kinases (GRKs), which limit opioid receptor activity. Gallein and M119 block G-dependent regulation of PLC and GRK2 without blocking GIRK or N-type Ca2+ channels and thereby biasing MORs toward pro-analgesic signaling. Gallein and M119 have been powerful probe compounds to validate the idea that pharmacological G blockade could improve the properties of opioid analgesics, but the chemical characteristics of the molecules makes them unsuitable for therapeutic development. Data with new chemical series’ derived from our high throughput screening (HTS) campaign support a G on-target mechanism of action. The primary goal of this application is to develop and mechanistically characterize novel “drug like” G inhibitors that can be utilized to improve the safety of opioid analgesics. This application is divided into 3 specific aims 1) We will diversify the chemistry of promising lead compounds derived from HTS with the goal of improving potency and “drug like” characteristics. We expect that upon completion of this aim we will identify a high potency “drug like” G inhibitor that can be a strong lead candidate for therapeutic development. 2) We will use whole animal PLC and -arrestin-2 knockout models, and brain slice electrophysiology to examine the roles of blocking feedback inhibition of MOR signaling by PLC, PKC and -arrestin pathways in the potentiating actions of G inhibitors. 3) There is a significant need to find an opioid analgesic for the treatment of chronic pain without abuse potential and adverse side effects. We will explore the utility of G inhibition in chronic opioid use in a mouse model of chronic inflammatory pain, with both chronic and acute administration and use a rat model of morphine self-administration.

该申请提出了一种通过受体后提高阿片类镇痛药安全性的新方法 G 蛋白  亚基的药理学靶向可改变 -阿片受体 (MOR) 的作用。实验室已鉴定出 G 蛋白  亚基的小分子抑制剂，其在体内功效与该应用相关的各种动物模型，我们鉴定了两种相关的 G 抑制剂，M119。和加仑可增加小鼠中 MOR 激动剂的镇痛效力，且不会增强副作用，包括出现耐受性、呼吸抑制、便秘或成瘾。阿片类镇痛药抑制剂有可能通过开放治疗途径来提高其安全性我们建议使用gallein 和M119 来修饰阿片类药物。通过阻断 MOR 下游的特定通路反馈同时保留促镇痛通路来发挥作用完整的 MOR 与 G 蛋白的 Gi 家族偶联，并通过 G 蛋白依赖性抑制促进镇痛。通过 G 依赖性调节 K+ (GIRK) 通道、N 型 Ca2+ 通道和抑制囊泡与突触膜的融合，同时 G 激活反馈通路。包括限制阿片受体的磷脂酶 C (PLC) 和 G 蛋白偶联受体激酶 (GRK) Gallein 和 M119 阻断 PLC 和 GRK2 的 G 依赖性调节，而不阻断 GIRK 或 N 型。 Ca2+ 通道，从而使 MOR 偏向促镇痛信号传导，Gallein 和 M119 非常强大。探针化合物验证药理学 G 阻断可以改善阿片类药物特性的观点镇痛药，但分子的化学特性使其不适合用于治疗新化学系列的开发数据来自我们的高通量筛选 (HTS) 活动。支持 G 目标作用机制该应用程序的主要目标是开发和实现。机制表征新型“类药”G 抑制剂，可用于提高阿片类药物的安全性该应用分为 3 个具体目标 1) 我们将使有前景的铅的化学成分多样化。我们期望从 HTS 中提取化合物，以提高效力和“类似药物”的特性。完成这一目标后，我们将确定一种高效的“类药物”G 抑制剂，它可以成为强有力的先导药物 2) 我们将使用整个动物 PLC 和 -arrestin-2 敲除模型，和脑切片电生理学来检查 PLC 阻断 MOR 信号传导反馈抑制的作用， PKC 和 -arrestin 通路在 G 抑制剂的增强作用中的作用 3) 迫切需要找到这种途径。一种用于治疗慢性疼痛的阿片类镇痛药，没有滥用潜力和不良副作用。探索 G 抑制在慢性炎性疼痛小鼠模型中慢性阿片类药物使用的效用，同时慢性和急性给药并使用吗啡自我给药的大鼠模型。