Non-Opioids for Inflammatory Pain: Adenylyl Cyclase 1 as a Novel Target

非阿片类药物治疗炎症性疼痛：腺苷酸环化酶 1 作为新靶点

• 批准号: 10525509
• 负责人: David L. Roman
• 金额: $ 6.01万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10525509
• 关键词: Active Sites; Adenine; Adenylate Cyclase; Antidepressive Agents; Automobile Driving; Binding; Binding Sites; Biochemical; Biological Assay; Calmodulin; Calmodulin 1; Cell physiology; Cellular Assay; Chemicals; Chronic inflammatory pain; Collaborations; DNA biosynthesis; Data; Development; Economic Burden; Emotional; Financial cost; Forskolin; Goals; Iowa; Knock-out; Knockout Mice; Legal; Libraries; Link; Location; Modeling; Neuronal Plasticity; Neurons; Non-Steroidal Anti-Inflammatory Agents; Opioid; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phenotype; Protein Isoforms; Public Health; Quality of life; Site; Structure; Tissues; Universities; adenylyl cyclase 1; chronic pain; chronic pain management; chronic painful condition; design; high throughput screening; improved; inflammatory pain; inhibitor; lead optimization; non-opioid analgesic; novel; novel strategies; opioid use; pain processing; pre-clinical; preclinical study; prevent; protein protein interaction; screening; side effect; small molecule inhibitor; social

ABSTRACT Chronic pain is a major concern in public health with financial costs projected to surmount $600 billon in the next year. Patients afflicted with chronic pain endure extreme emotional, physical, and social burdens, resulting in severely diminished quality of life. Unfortunately, drugs currently used for chronic pain management, such as NSAIDs, opioids, neuronal stabilizers, and antidepressants, do not typically provide sufficient relief to restore full quality of life, and in many instances these treatments themselves limit patients, such as opioid treatment preventing a patient from legally driving. Recent preclinical studies have identified neuronal adenylyl cyclase type 1 (AC1) as a novel target for treating chronic pain. AC1 is highly expressed in neuronal tissues associated with pain processing and neuronal plasticity, and studies using AC1 knockout mice provide direct evidence linking AC1 to chronic inflammatory pain conditions. Furthermore, AC1 inhibitors would lack the side effects associated with other agents (e.g. opioids) used to treat chronic inflammatory pain. The development of AC1 inhibitors represents a unique challenge, as demonstrated by a prior preclinical AC1 inhibitor, NB001. NB001 has significant shortcomings, including modest selectivity over other adenylyl cyclase isoforms, likely due to its adenine-like structure. Compounds of this type are called P-site inhibitors and act by binding to the active site of AC that is conserved among all isoforms. Additional concerns for adenine-containing molecules like NB001 include effects on other cellular processes such as DNA synthesis. We hypothesize that developing a small molecule inhibitor of AC1 will allow us to mimic the AC1 knockout phenotype and provide a new avenue for the treatment of chronic inflammatory pain. We designed our studies to target NOT the conserved P-site or forskolin-binding site, but rather a novel approach, targeting the unique protein-protein interaction of AC1 and calmodulin (CaM). AC1 and AC8 are both activated by CaM, however, the CaM binding domains are unique in structure and location providing an unprecedented opportunity to achieve AC1 selectivity. Thus, the goals of this proposal are to: 1) develop a novel AC1/CaM biochemical screening assay, 2) implement this novel assay in a high throughput screen to interrogate a library of 100,000 compounds for inhibitors of the AC1/CaM protein- protein interaction, and 3) validate and chemically optimize lead molecules using cellular assays focused on selectivity and potency to guide medicinal chemistry efforts. To date, we have completed initial studies to develop the novel screening assay, established a subset of the necessary assays, and cemented the collaboration between the University of Iowa and Purdue University for the successful completion of our aims. We anticipate the identification of selective AC1 inhibitors that ultimately be improved and applied in models of chronic inflammatory pain.

抽象的 慢性疼痛是公共卫生领域的一个主要问题，预计其财务成本将超过 6000 亿美元 明年。患有慢性疼痛的患者承受着极端的情感、身体和社会负担， 导致生活质量严重下降。不幸的是，目前用于治疗慢性疼痛的药物 管理，如非甾体抗炎药、阿片类药物、神经稳定剂和抗抑郁药，通常不提供 足以恢复生活质量的缓解，并且在许多情况下，这些治疗本身限制了 患者，例如阿片类药物治疗阻止患者合法驾驶。最近的临床前研究已 将神经元腺苷酸环化酶 1 型 (AC1) 确定为治疗慢性疼痛的新靶标。 AC1 是高度 在与疼痛处理和神经元可塑性相关的神经元组织中表达，并使用 AC1 基因敲除小鼠提供了 AC1 与慢性炎症性疼痛相关的直接证据。 此外，AC1 抑制剂不会产生与其他药物（例如阿片类药物）相关的副作用。 治疗慢性炎症疼痛。 AC1 抑制剂的开发是一个独特的挑战，因为 先前的临床前 AC1 抑制剂 NB001 已证明这一点。 NB001有明显的缺点，包括 与其他腺苷酸环化酶亚型相比具有适度的选择性，可能是由于其腺嘌呤样结构。化合物 这种类型的抑制剂称为 P 位点抑制剂，通过与所有类型中保守的 AC 活性位点结合而发挥作用。 同工型。对 NB001 等含腺嘌呤分子的其他担忧包括对其他细胞的影响 DNA 合成等过程。我们假设开发 AC1 小分子抑制剂将 使我们能够模仿 AC1 敲除表型，并为治疗慢性病提供新途径 炎症性疼痛。我们设计的研究不是针对保守的 P 位点或毛喉素结合位点，而是 这是一种新颖的方法，针对 AC1 和钙调蛋白 (CaM) 独特的蛋白质-蛋白质相互作用。交流1 和 AC8 均由 CaM 激活，但 CaM 结合域结构独特， 位置提供了前所未有的机会来实现 AC1 选择性。因此，本提案的目标 我们的目标是：1) 开发一种新型 AC1/CaM 生化筛选测定法，2) 在高通量中实施这种新型测定法 通量筛选，用于询问 100,000 种化合物库中的 AC1/CaM 蛋白抑制剂 - 蛋白质相互作用，以及 3) 使用专注于以下方面的细胞测定来验证和化学优化先导分子 指导药物化学工作的选择性和效力。迄今为止，我们已经完成了初步研究 开发新的筛选试验，建立必要试验的子集，并巩固 爱荷华大学和普渡大学之间的合作成功完成了我们的 目标。我们预计选择性 AC1 抑制剂的鉴定最终将得到改进并应用于 慢性炎性疼痛模型。