Improved Therapeutics for the Resurrection of the Aged Form of Acetylcholinesterase

老化乙酰胆碱酯酶复活的改进疗法

• 批准号: 10238898
• 负责人: Christopher M. Hadad
• 金额: $ 37.07万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10238898
• 关键词: Acetylcholine; Acetylcholinesterase; Active Sites; Aging; Ammonium; Back; Binding; Binding Sites; Biochemical; Chemical Agents; Chemical Warfare Agents; Chemicals; Cleaved cell; Complement; Computing Methodologies; Country; Crystallization; Docking; Enzymes; Event; Exposure to; Family; Human; In Vitro; Kinetics; Lead; Libraries; Ligands; Mechanics; Methods; Military Personnel; Molecular; Neuromuscular Junction; Organic Synthesis; Organophosphorus Compounds; Oximes; Peripheral; Pesticides; Pharmaceutical Preparations; Process; Property; Proteomics; Reporting; Serine; Societies; Soman; Structure; Testing; Therapeutic; Variant; adduct; age effect; aged; cholinergic; design; humanized mouse; improved; in silico; in vivo; mass casualty; medical countermeasure; methylphosphonate; molecular dynamics; mouse model; nerve agent; novel; quantum; quinone methide; receptor; scaffold; screening; small molecule libraries; success; toxicant

One of the major deficiencies of current medical countermeasures is that current oximes cannot reactivate nerve agent-inhibited acetylcholinesterase (AChE) that has aged after exposure to organophosphorus G- and V-type chemical nerve agents. Our team has demonstrated the first, and only, compounds that have the capability to “resurrect” in vitro the methylphosphonate-aged form of AChE to an active, native state. We posit that this resurrection accomplishes two distinct steps – first, to realkylate the anionic aged form back to a neutral, phosphylated (inhibited) serine residue and then to reactivate the inhibited form back to the native AChE. This proposal focuses on expanding on these successful chemical frameworks in order to identify even more efficacious drug-like molecules that will enable the aged form of AChE to be resurrected in vivo. Using various quinone methide precursor (QMP) frameworks, we will use computational and experimental approaches to prepare a chemical library and then to screen these compounds for efficacy in resurrecting the aged form of AChE back to native activity. There are no approved countermeasures that can resurrect the aged form of AChE to its active form; however, resurrection of aged AChE is the holy grail against OP exposure. If realkylation of aged AChE can occur, then AChE can be fully rejuvenated as oximes (and other reactivators) exist that are potent nucleophiles for cleaving the O–P bond of the phosphylated serine, thereby reforming active AChE. Thus, the objective of this proposal is to expand on our successful chemical frameworks to react selectively with aged AChE to form stable alkylphosphonate-AChE adducts that can then be reactivated to the native state, thereby reversing the effects of aging by chemical nerve agents. The design of alkylating compounds will be guided by state-of-the-art computational methods (molecular dynamics, molecular docking, and quantum mechanical methods) that predict the ligand-receptor interactions of alkylating compounds with aged AChE. Using in silico guidance, libraries of alkylating quinone methide precursor (QMP) compounds will be synthesized, and then tested in a kinetic screening process, and complemented by mass spectrometric and proteomic studies. The best lead compounds will be evaluated for their in vitro drug-like properties and tested in vivo in a humanized mouse model for AChE.

当前医学对策的主要缺陷之一是现有的肟类药物不能 神经毒剂抑制的乙酰胆碱酯酶(AChE)在暴露于 有机磷G型和V型化学神经毒剂。我们的团队已经展示了第一个，并且 只有那些有能力在体外使甲基膦酸类药物复活的化合物 将疼痛的形式转化为活动的、自然的状态。我们假设这种复活完成了两个截然不同的步骤 -首先，将阴离子陈化形式重新烷基化为中性的、磷化(抑制)的丝氨酸残基，以及 然后将被抑制的形式重新激活回本地AChE。这项提案的重点是扩大 这些成功的化学框架，以识别更有效的类药物分子， 将使老化的AChE形式在体内复活。使用各种不同的苯醌甲烷前驱体 (QMP)框架，我们将使用计算和实验方法来制备一种化学品 文库，然后筛选这些化合物在使老化的疼痛形式死灰复燃 原生活动。 没有被批准的对策可以使老化的AChE恢复到其活跃的形式； 然而，老年疼痛的复苏是防止OP暴露的圣杯。老年AChE的If再烷基化 可以发生，然后AChE可以完全恢复活力，因为存在有效的肟类(和其他复活剂) 亲核剂，用于裂解磷酸化丝氨酸的O-P键，从而重整活性AChE。 因此，这项提议的目标是扩大我们成功的化学反应框架。 选择性地与陈化的AChE形成稳定的烷基膦酸-AChE加合物，然后可以 重新激活到自然状态，从而逆转化学神经毒剂老化的影响。 烷基化化合物的设计将由最先进的计算方法(分子 动力学、分子对接和量子力学方法)来预测配体-受体 烷基化化合物与老年乙酰胆碱酯酶的相互作用。烷基化库在硅胶制导中的应用 将合成甲基苯二酚前体化合物，然后在动力学筛选中进行测试 过程，并辅之以质谱学和蛋白质组学研究。最好的先导化合物 将评估它们的体外类药物特性，并在体内人源化小鼠模型中进行测试 疼死了。