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: 10405130
• 负责人: David L. Roman
• 金额: $ 40.9万
• 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/10405130
• 关键词: Active Sites; Adenine; Adenylate Cyclase; Antidepressive Agents; Arthritis; Automobile Driving; Binding; Binding Sites; Biochemical; Biological Assay; Calcium; Calmodulin; Calmodulin 1; Catalogs; Cell membrane; Cell physiology; Cells; Cellular Assay; Chelating Agents; Chemicals; Chronic inflammatory pain; Chronic low back pain; Collaborations; Complex; DNA biosynthesis; Data; Development; Drug Industry; Economic Burden; Egtazic Acid; Emotional; Financial cost; Fluorescence Polarization; Forskolin; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Genetic study; Goals; Health; Iowa; Knock-out; Knockout Mice; Legal; Libraries; Link; Location; Modeling; Mus; Neurobiology; Neuronal Plasticity; Neurons; New Agents; Non-Steroidal Anti-Inflammatory Agents; Opiate Addiction; Opioid; Pain; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phenotype; Protein Isoforms; Public Health; Quality of life; Research Personnel; Risk; Role; Signal Transduction; Site; Specificity; Stimulus; Structure; Therapeutic; Tissues; Universities; War; addiction; adenylyl cyclase 1; base; cancer pain; cheminformatics; chronic pain; chronic pain management; chronic painful condition; complement C2b; cost; cyanine dye 5; design; high throughput screening; improved; in vitro Model; in vivo; inflammatory pain; inhibitor; knockout animal; lead optimization; new therapeutic target; non-opioid analgesic; novel; novel strategies; opioid epidemic; opioid use; pain processing; pre-clinical; preclinical study; prevent; protein protein interaction; scaffold; screening; side effect; small molecule; small molecule inhibitor; social; synthetic peptide

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的主要关注点，预计将为600美元的亿万美元的经济成本关注 明年。患有慢性疼痛的患者忍受了极端的情感，身体和社会负担， 导致生活质量严重降低。不幸的是，目前用于慢性疼痛的药物 NSAID，阿片类药物，神经元稳定剂和抗抑郁药等管理通常不提供 足够的救济以恢复全部生活质量，在许多情况下，这些治疗本身都限制了 患者，例如阿片类药物治疗，阻止患者合法驾驶。最近的临床前研究有 鉴定出神经元腺苷酸环化酶1型（AC1）是治疗慢性疼痛的新靶标。 AC1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敲除小鼠提供了直接证据，将AC1与慢性炎症性疼痛条件联系起来。 此外，AC1抑制剂将缺乏与其他因素（例如阿片类药物）相关的副作用 治疗慢性炎症疼痛。 AC1抑制剂的发展代表了一个独特的挑战，因为 由先前的临床前AC1抑制剂NB001证明。 NB001有很大的缺点，包括 与其他腺苷环酶同工型相比，适度的选择性可能是由于其腺嘌呤样结构所致。化合物 这种类型的称为P位点抑制剂，并通过与AC的活性位点结合而起作用 同工型。含腺嘌呤的分子（如NB001）的其他问题包括对其他细胞的影响 诸如DNA合成之类的过程。我们假设开发AC1的小分子抑制剂将 允许我们模仿AC1敲除表型，并为治疗慢性提供新的途径 炎症性疼痛。我们设计的研究不是针对保守的P位点或福斯科林结合部位，而是针对的，而是针对 而不是一种新颖的方法，它针对AC1和钙调蛋白（CAM）的独特蛋白质蛋白质相互作用。 AC1 但是 位置为实现AC1选择性提供了前所未有的机会。因此，该提议的目标 为：1）开发一种新颖的AC1/CAM生化筛选测定法，2）以高 吞吐量屏幕以询问100,000种化合物的库，以供AC1/CAM蛋白的抑制剂 蛋白质相互作用，以及3）使用专注于细胞测定法对铅分子进行验证和化学优化铅分子 选择性和效力指导药物化学工作。迄今为止，我们已经完成了初步研究 开发新颖的筛选测定法，确定必要测定的子集，并巩固 爱荷华大学和普渡大学之间的合作成功完成了 目标。我们预计最终改进和应用的选择性AC1抑制剂的鉴定 慢性炎症疼痛的模型。