THERAPEUTIC PROFILES OF OPIOID DELTA RECEPTORS--NEW MODALITIES

阿片类药物 Delta 受体的治疗概况——新模式

• 批准号: 6104007
• 负责人: Frank Porreca
• 金额: $ 10.45万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6104007
• 关键词: analgesics; cell line; cell type; dorsal root; drug addiction; drug design /synthesis /production; drug screening /evaluation; fluorescence microscopy; laboratory rat; narcotic antagonists; neuropeptide receptor; neuropeptides; oligonucleotides; opioid receptor; peptide analog; pharmacokinetics; receptor expression; receptor sensitivity

The aims of this project are based on three hypothesis. Hypothesis 1 is that the opioid delta receptor may exist as subtypes. An antisense strategy will be employed in NG 108-15 cells, in primary DRG cultures and in an antinociceptive model in vivo. Antisense oligodeoxynucleotide (ODN) "knock-down" of delta receptors in NG108-15 cells will be studied using ODNs directed at the DOR-1 clone and at a conserved sequence in the opioid mu, delta and kappa receptors (conserved opioid receptor; COR). Cellular uptake and ODN translocation, time-course, concentration-dependence and inhibition of the delta receptor expression will be determined with fluorescence microscopy and radioligand binding; control ODN sequences will be emphasized. In DRG cultures DOR or COR antisense ODN will be used to evaluate ODN uptake and morphological distribution and to validate the DOR ODN selectivity against delta mu and kappa receptors. In rats, DOR/COR ODNs will be used to elucidate the distribution of spinal ODNs and "knock-down" of opioid receptors and possible differential sensitivity of putative delta subtype selective agonists (but not mu or kappa agonists), to DOR-1 antisense ODN effects as well as time-course and recovery of the antinociceptive effect. Hypothesis 2 is that activation of supraspinal opioid delta receptors will produce antinociception not necessarily dependent on the descending pathway characterized for opioid mu receptor agonists. Localized microinjection of opioid subtype selective agonists and antagonists into selected supraspinal loci will be employed to establish receptor selective antinociceptive activity. The involvement of descending pathways following activation of confirmed supraspinal delta antinociception will be studied by examining suppression of nociceptive induced behavior and c-FOS expression in the spinal cord in control rats, or in animals with lesions of the dorsolateral funiculus (DLF). Hypothesis 3 is that activation of both supraspinal and spinal opioid delta receptors will result in a synergistic antinociceptive effect. Analysis of antinociception elicited by supraspinal/spinal co-administration of subtype-selective agonists in a fixed-ratio paradigm with isobolographic methods will be used. These experiments should yield significant new information about the role of the delta receptor in nociception and the bossibility of subtypes of this receptor in vitro and in vivo.

这个项目的目标是基于三个假设。 假设1是阿片δ受体可能以 亚型 NG 108-15将采用反义策略 细胞，在原代DRG培养物和抗伤害感受模型中 in vivo. 反义寡脱氧核苷酸（ODN）“敲低” 将使用ODNs研究NG 108 -15细胞中的δ受体 针对DOR-1克隆和DOR-1中的保守序列， 阿片样物质μ、δ和κ受体（保守的阿片样物质受体; COR）。 细胞摄取和ODN易位，时间过程， δ受体的浓度依赖性和抑制 用荧光显微镜测定表达， 放射性配体结合;控制ODN序列将被强调。 在DRG培养物中，DOR或COR反义ODN将用于评估 ODN摄取和形态分布，并验证DOR ODN对δ μ和κ受体的选择性。 在大鼠中， DOR/COR ODNs将用于阐明脊髓中DOR/COR的分布。 ODNs和阿片受体的“敲低”以及可能的 推定δ亚型选择性的差异敏感性 激动剂（但不是mu或kappa激动剂）与DOR-1反义ODN 影响以及时间进程和恢复的 抗伤害效应 假设2： 脊髓上阿片δ受体将产生抗伤害感受 不一定依赖于下行通路， 阿片样μ受体激动剂 局部微量注射 阿片类亚型选择性激动剂和拮抗剂， 脊髓上位点将用于建立受体选择性 抗伤害活性。 下行通路的参与 在确认棘上三角肌激活后， 抗伤害感受将通过检查 伤害性行为与脊髓c-FOS表达 在对照大鼠中，或在具有损伤的动物中， 背外侧索（DLF）。 假设3：激活 脊髓上和脊髓阿片δ受体的作用 具有协同抗伤害作用。 分析 脊髓上/脊髓联合给药引起的抗伤害感受 亚型选择性激动剂在一个固定比例的范例， 将使用等辐射线方法。 这些实验应该 关于三角洲作用的重要新信息 受体在伤害感受和bossibility亚型这一点 受体在体外和体内。