Microtubule affinity-regulating kinase (MARK4) plays a key role in Alzheimer's disease (AD) development as its overexpression is directly linked to increased tau phosphorylation. MARK4 is a potential drug target of AD and is thus its structural features are employed in the development of new therapeutic molecules. Donepezil (DP) and rivastigmine tartrate (RT) are acetylcholinesterase (AChE) inhibitors and are used to treat symptomatic patients of mild to moderate AD. In keeping with the therapeutic implications of DP and RT in AD, we performed binding studies of these drugs with the MARK4. Both DP and RT bound to MARK4 with a binding constant (K) of 107 M-1. The temperature dependency of binding parameters revealed MARK-DP complex to be guided by static mode while MARK-RT complex to be guided by both static and dynamic quenching. Both drugs inhibited MARK4 with IC50 values of 5.3 μM (DP) and 6.74 μM (RT). The evaluation of associated enthalpy change (ΔH) and entropy change (ΔS) implied the complex formation to be driven by hydrogen bonding making it seemingly strong and specific. Isothermal titration calorimetry further advocated a spontaneous binding. In vitro observations were further complemented by the calculation of binding free energy by molecular docking and interactions with the functionally-important residues of the active site pocket of MARK4. This study signifies the implications of AChE inhibitors, RT, and DP in Alzheimer's therapy targeting MARK4.

中文翻译：

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AChE抑制剂Donepezil和酒石酸Rivastigmine抑制MARK4：阿尔茨海默氏病疗法的见解。

微管亲和力调节激酶（MARK4）在阿尔茨海默氏病（AD）的发展中起关键作用，因为其过表达与tau磷酸化的增加直接相关。MARK4是AD的潜在药物靶标，因此其结构特征被用于开发新的治疗分子。多奈哌齐（DP）和酒石酸卡巴拉汀（RT）是乙酰胆碱酯酶（AChE）抑制剂，用于治疗轻度至中度AD的有症状患者。为了与DP和RT在AD中的治疗意义保持一致，我们对这些药物与MARK4进行了结合研究。DP和RT均以107 M-1的结合常数（K）结合至MARK4。结合参数的温度依赖性表明，MARK-DP复合物受静态模式控制，而MARK-RT复合物受静态和动态猝灭控制。两种药物均抑制MARK4，IC50值分别为5.3μM（DP）和6.74μM（RT）。对相关的焓变（ΔH）和熵变（ΔS）的评估暗示了氢键驱动的复合物形成，使其看起来牢固而特异。等温滴定量热法进一步提倡自发结合。通过分子对接以及与MARK4活性位点口袋中功能上重要的残基的相互作用计算结合自由能，进一步补充了体外观察结果。这项研究表明，AChE抑制剂，RT和DP在阿尔茨海默氏病治疗MARK4的意义。对相关的焓变（ΔH）和熵变（ΔS）的评估暗示了氢键驱动的复合物形成，使其看起来牢固而特异。等温滴定量热法进一步提倡自发结合。通过分子对接以及与MARK4活性位点口袋中功能上重要的残基的相互作用计算结合自由能，进一步补充了体外观察结果。这项研究表明，AChE抑制剂，RT和DP在阿尔茨海默氏病治疗MARK4的意义。对相关的焓变（ΔH）和熵变（ΔS）的评估暗示了氢键驱动的复合物形成，使其看起来牢固而特异。等温滴定量热法进一步提倡自发结合。通过分子对接以及与MARK4活性位点口袋中功能上重要的残基的相互作用计算结合自由能，进一步补充了体外观察结果。这项研究表明AChE抑制剂，RT和DP在阿尔茨海默氏病治疗MARK4的意义。通过分子对接以及与MARK4活性位点口袋中功能上重要的残基的相互作用计算结合自由能，进一步补充了体外观察结果。这项研究表明，AChE抑制剂，RT和DP在阿尔茨海默氏病治疗MARK4的意义。通过分子对接以及与MARK4活性位点口袋中功能上重要的残基的相互作用计算结合自由能，进一步补充了体外观察结果。这项研究表明AChE抑制剂，RT和DP在阿尔茨海默氏病治疗MARK4的意义。