Improved Therapeutics for the Resurrection of the Aged Form of Acetylcholinesterase

当前医疗对策的主要缺陷之一是当前的肟不能 重新激活暴露于神经毒剂后老化的乙酰胆碱酯酶（AChE） 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 仅具有在体外“复活”磷酸甲酯老化能力的化合物 AChE 形式转变为活性的天然状态。我们假设这次复活完成了两个不同的步骤 – 首先，将阴离子老化形式重新烷基化回中性、磷酸化（抑制）丝氨酸残基， 然后将受抑制的形式重新激活为天然 AChE。该提案的重点是扩大 这些成功的化学框架是为了识别更有效的类药物分子 将使老化形式的 AChE 在体内复活。使用各种醌甲基化物前体 （QMP）框架，我们将使用计算和实验方法来制备化学物质 文库，然后筛选这些化合物在复活老化形式的 AChE 方面的功效 本机活动。 没有经过批准的对策可以使老化形式的乙酰胆碱酯酶恢复到其活性形式； 然而，老化的 AChE 的复活是对抗 OP 暴露的圣杯。如果老化的AChE重新烷基化 可以发生，然后 AChE 可以完全恢复活力，因为存在有效的肟（和其他重激活剂） 亲核试剂用于裂解磷酸化丝氨酸的 O-P 键，从而重组活性 AChE。 因此，该提案的目标是扩展我们成功的化学框架以进行反应 选择性地与老化的 AChE 形成稳定的烷基膦酸酯-AChE 加合物，然后可以 重新激活到天然状态，从而逆转化学神经毒剂造成的衰老影响。 烷基化化合物的设计将以最先进的计算方法（分子计算方法）为指导 动力学、分子对接和量子力学方法）预测配体-受体 烷基化化合物与老化乙酰胆碱酯酶的相互作用。使用计算机指导、烷基化库 将合成醌甲基化物前体（QMP）化合物，然后进行动力学筛选测试 过程，并辅以质谱和蛋白质组学研究。最好的铅化合物 将对其体外药物特性进行评估，并在人源化小鼠模型中进行体内测试 乙酰胆碱酯酶。

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.