Pharmacology of Kappa Opioid Receptor

Kappa 阿片受体的药理学

基本信息

批准号：
10212993
负责人：
LEE-YUAN LIU-CHEN
金额：
$ 53.9万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10212993
关键词：
ADRBK2 gene Abbreviations Abnormal coordination Absence of pain sensation Adenylate Cyclase Agonist Analgesics Anhedonia Animal Model Antibodies Antipruritic Effect Antipruritics Arrestins Attenuated Behavior Behavioral Biochemical Pharmacology Brain Brain region Cells Cellular biology Clinical Coupled Development Diuresis Dose Epidemic FRAP1 gene Familiarity Family G Protein-Coupled Receptor Signaling G protein coupled receptor kinase G-Protein-Coupled Receptors GRK5 gene GRK6 gene GTP-Binding Proteins Hemodialysis Heroin Abuse Immunoblotting In Vitro Japan Lead Ligands Light MAP Kinase Gene MAPK3 gene Measures Mediating Mental Depression Molecular Motor Mus Mutant Strains Mice Mutation Names Opioid Analgesics Opioid Receptor Pathway interactions Patients Penetration Pharmaceutical Preparations Pharmacology Phase Phosphorylation Potassium Channel Problem Solving Proteins Pruritus Reporting Rhodopsin Role Sedation procedure Signal Pathway Signal Transduction Structure System Techniques Testing Therapeutic Therapeutic Effect Time Translating Ventilatory Depression Water base behavioral response bench to bedside beta-arrestin desensitization dysphoria in vivo kappa opioid receptors natural hypothermia novel opioid abuse opioid use p38 Mitogen Activated Protein Kinase phosphoproteomics prescription opioid abuse receptor receptor internalization recruit sedative side effect

项目摘要

Opioid receptors (µ, d and k), are Gi/o-coupled, rhodopsin-like receptors. κ opioid receptor (KOPR) agonists may be useful as analgesics and antipruritic agents without abuse potential and respiratory depression associated with currently use µ opioid analgesics. However, prototypic selective KOPR agonists cause dysphoria or aversion, which limits their development. G protein-coupled receptors (GPCRs) signal via both G protein or arrestin to activate different downstream effectors. Biased agonists preferentially activate G protein- or arrestin-mediated signaling and thus may have advantages over balanced or unbiased agonists in that they may produce therapeutic effects with fewer side effects. However, translating in vitro ligand bias to in vivo pharmacology has been uncertain. Nalfurafine, the only selective KOPR agonist in clinical use, is prescribed in Japan for treatment of uremic pruritus, without causing dysphoria at therapeutic doses. In mice, we observed that nalfurafine caused conditioned place aversion (CPA) at doses higher than the effective doses for the antinociceptive and anti-scratch effects; however, the reverse was true for two other selective KOPR agonists, U50,488H and MOM-SalB. Similarly, U50,488H, but not nalfurafine, induced anhedonia. Thus, we established an animal model to understand the mechanisms underlying separation of anti-pruritic and analgesic effects from dysphoria / aversion of KOPR agonists. Importantly, we found in mouse brains U50,488H and MOM-SalB caused robust KOPR phosphorylation, but nalfurafine did not. Also,U50,488H, but not nalfurafine, enhanced phosphorylation of some proteins downstream of mTOR and the mTOR pathway may be involved in KOPR-mediated CPA. For Specific Aim 1, We will test the hypothesis that the ability of agonists to promote CPA is related to its ability cause KOPR phosphorylation by examining several structurally distinct KOPR agonists. KOPR phosphorylation will be detected with immunoblotting using our own antibodies that specifically recognize phosphorylated KOPR. For Specific Aim 2, we will examine the differences between U50,488H and nalfurafine in downstream phosphoproteomic changes in brain regions important in KOPR pharmacology. Furthermore, we will investigate the involvement of differentially regulated proteins / pathways in KOPR-mediated CPA and anhedonia. For Specific Aim 3, we will generate mutant mouse lines to examine the roles of agonist-promoted KOPR phosphorylation and GRK5 and GRK6 in KOPR pharmacology in vivo. KOPR-mediated antipruritic, antinociceptive, aversive and sedative effects and motor incoordination will be used as the in vivo pharmacological measures. Our “from bedside to bench” approach, distinctly different from the commonly used “from bench to bedside” strategy, allows us to circumvent the challenges of translating in vitro cell-based results to in vivo pharmacology. Taken together, the proposed studies will greatly advance our understanding of KOPR pharmacology at the molecular, cellular, and behavioral levels and signaling at a system level. Signaling pathways identified to be involved in KOPR-mediated aversion and anhedonia may shed light on mechanisms underlying aversion- and depression-like behaviors in general. In addition, it may lead to development of KOPR agonists that cause lower dysphoria and can be used as anti-itch medications and analgesics, which will contribute to solving the problems of the opioid abuse epidemic.

阿片受体（µ，d和k）是gi/o偶联的类似视紫红质的受体。 κ阿片受体（KOPR）激动剂可能有用作为镇痛药和抗乳脂药，没有滥用潜力和与当前使用µ相关的呼吸抑郁症阿片类镇痛药。但是，原型的选择性Kopr激动剂会引起烦躁不安或厌恶，这限制了它们的发展。 G蛋白偶联受体（GPCR）通过G蛋白或逮捕蛋白信号激活不同的下游效果。有偏的激动剂优先激活G蛋白或阻滞蛋白介导的信号传导，因此可能具有优势过度平衡或公正的激动剂可能会产生副作用较少的热效应。然而，将体外配体偏置转化为体内药理学的偏差尚不确定。 Nalfurafine，唯一的选择性Kopr agonist 在临床用途中，在日本处方用于治疗尿毒症瘙痒，而不会在治疗剂量时引起烦躁不安。在小鼠，我们观察到纳尔列芬在以高于有效剂量的剂量高于有效剂量的情况抗伤害感受和抗抓症；但是，对于另外两个选择性的Kopr激动剂而言，相反的相反， U50,488H和妈妈 - 萨尔布。同样，U50,488H，但不是Nalfurafine诱发了Anhedonia。那，我们建立了动物了解抗pruritic和镇痛作用与吞咽困难 /厌恶的抗脑症和镇痛作用的基础机制科普激动剂。重要的是，我们在小鼠大脑U50,488H中发现，妈妈 - 萨尔布引起了强大的Kopr 磷酸化，但纳尔列芬没有。另外，U50,488H，但不是Nalfurafine，某些蛋白质的磷酸化增强 MTOR和MTOR途径的下游可能与KOPR介导的CPA有关。对于特定目标1，我们将检验以下假设：激动剂促进CPA的能力与其能力有关，导致Kopr磷酸化。检查几种结构上不同的Kopr激动剂。通过免疫印迹检测KOPR磷酸化使用我们自己的抗体，这些抗体专门识别磷酸化的Kopr。对于特定目标2，我们将检查 U50,488H和Nalfurafine在大脑区域的下游磷酸蛋白质组学变化中的差异很重要 KOPR药理学。此外，我们将研究不同调节的蛋白质 /途径的参与 KOPR介导的CPA和Anhedonia。对于特定的目标3，我们将生成突变的小鼠线以检查在体内KOPR药理学中，激动剂促进的Kopr磷酸化和GRK5和GRK6。 KOPR介导的抗毒性，抗伤害感受，厌恶和镇静作用以及运动不协调将用作体内药理措施。我们的“从床边到长凳”的方法，与常用的“从长凳到床边”策略，使我们能够规避将基于体外细胞的结果转化为体内的挑战药理。综上所述，拟议的研究将大大提高我们对KOPR药理学的理解分子，细胞和行为水平以及系统水平的信号传导。确定涉及的信号通路在Kopr介导的厌恶和Anhedonia中，可能会揭示出厌恶和抑郁状的机制一般行为。此外，它可能导致kopr激动剂的发展，引起较低的烦躁不安，可以是用作抗核药物和镇痛药，这将有助于解决阿片类药物滥用流行病的问题。