Assay development for characterization of Adrb3 antagonists as pain therapeutics

项目摘要 功能性疼痛综合征（FPS）影响超过1亿人，但传统的治疗方法仍然无效。 药物治疗，如阿片类药物，疗效差，有不良的中枢副作用。我们的长期 我们的目标是为FPS患者开发更安全，更有效的镇痛药，特别是外周限制性镇痛药 β-3肾上腺素能受体（Adrb3）的拮抗剂。Adrb3受体是G蛋白偶联受体， 被儿茶酚胺激活在临床研究中，我们确定患有慢性FPS的患者，例如 纤维肌痛、下背痛和肠易激综合征会增加儿茶酚胺水平， 儿茶酚-O-甲基转移酶（COMT;一种代谢儿茶酚胺的酶）水平降低。 与临床症状一致，我们已经表明，在啮齿类动物中，COMT的药理学抑制产生 通过激活外周Adrb3引起身体多个部位疼痛。疼痛是由外周脂肪细胞Adrb3- 介导的局部组织中促炎细胞因子的增加，并通过随后的 脊髓组织中促炎细胞因子和脊髓组织中促分裂原活化蛋白激酶（MAPK）的活化 感受疼痛的伤害感受器的细胞体和中央末梢。因此，Adrb3是一个新的和有吸引力的目标， 治疗慢性功能性疼痛和疼痛相关炎症。然而，现有的工具化合物 缺乏对Adrb 3的选择性或代谢特性较差。为了成功地建立一个强大的药物发现 Abrb3拮抗剂的平台，在这项提案中，我们将开发和验证一组体外，基于细胞的 本发明的目的在于提供用于充分表征新型Adrb3配体的药理学的测定。高通量的发展， 基于板的测定对于准确评价新化合物的亲和力、效力、功效、选择性 和目标验证，作为长期药物化学活动的一部分，寻求生产新的镇痛药 具有改善的特异性和副作用特征，用于治疗功能性疼痛综合征。

PROJECT SUMMARY Functional pain syndromes (FPS) affect over 100 million people, yet remain ineffectively treated by conventional pharmacotherapies, such as opioids, that have poor efficacy and adverse central side effects. Our long-term goal is to develop safer, more effective analgesics for patients with FPS, specifically peripherally-restricted antagonists of the beta-3 adrenergic receptor (Adrb3). The Adrb3 receptor is a G protein-coupled receptor that is activated by catecholamines. In clinical studies, we determined that patients with chronic FPS such as fibromyalgia, low back pain, and irritable bowel syndrome have increased levels of catecholamines alongside reduced levels of catechol-O-methyltransferase (COMT; an enzyme that metabolizes catecholamines). Consistent with clinical syndromes, we have shown that pharmacologic inhibition of COMT in rodents produces pain at multiple body sites via activation of peripheral Adrb3. The pain is initiated by peripheral adipocyte Adrb3- mediated increases in pro-inflammatory cytokines in local tissues and maintained by subsequent increases in pro-inflammatory cytokines in spinal tissues and activation of mitogen activated protein kinases (MAPKs) in the cell bodies and central terminals of pain-sensing nociceptors. Thus, Adrb3 is a novel and attractive target for the treatment of chronic functional pain and pain-relevant inflammation. However, existing tool compounds either lack selectivity for Adrb3 or have poor metabolic properties. To successfully build a robust drug discovery platform for Abrb3 antagonists, in this proposal, we will develop and validate a battery of in vitro, cell-based assays to fully characterize the pharmacology of novel Adrb3 ligands. The development of high throughput, plate-based assays is critical for accurate evaluation of novel compound affinity, potency, efficacy, selectivity, and target validation, as part of a long-term medicinal chemistry campaign that seeks to produce new analgesics with improved specificity and side-effect profiles for the treatment of functional pain syndromes.