Acetylcholinesterase (AChE) inhibition is a significant strategy for preventing Alzheimer's disease (AD) and neurodegenerative diseases. In this study, a dereplication system was utilized to rapidly identify and characterize acetylcholinesterase-interacting compounds by comparing UPLC-MS/MS profile screening approach and molecular docking analysis, derived from the extracts and fractions of the stem of (Sims) Little. Eleven potential AChE inhibitors were isolated and identified from the ethyl acetate extract of , including an undescribed alkaloid (), namely glybomine D, eight known alkaloids (), a flavonoid (), and a phytosterol (). The inhibitory potential of these compounds against AChE was assessed, with -methylglycosolone (), 1,3-dimethoxy-2-hydroxy-10-methyl-9(10)-acridinone (), skimmianine () and arborine (), regarded as effective inhibitors, yielding IC values of 39.81 μM, 41.53 μM, 49.40 μM, and 59.92 μM, respectively. Notably, -methylglycosolone exhibited the highest potency. Four of these potent AChE inhibitors exhibited mixed-type inhibition. However, -methylglycosolone (), 1,3-dimethoxy-2-hydroxy-10-methyl-9(10)-acridinone (), and arborine () were first reported modulating with acetylcholinesterase activity. Furthermore, molecular docking revealed -methylglycosolone () superior binding affinity (–23.749 kcal/mol) compared to other compounds, mainly by interacting with the peripheral anionic site of AChE, which forms hydrogen bonds and hydrophobic forces may play an important role, interaction with amino acid residues such as Tyr341, Tyr72, Ser293, and Arg296 in the active cavity, which is crucial for effective and selective inhibition of AChE activity. ADMET predictions suggest that arborine (), skimmianine (), and -methylglycosolone () demonstrate favorable permeability across the blood–brain barrier, while 1,3-dimethoxy-2-hydroxy-10-methyl-9(10)-acridinone () exhibits comparatively reduced permeability. These findings highlight the potential of these compounds as natural AChE inhibitors for treating neurodegenerative diseases.

中文翻译：

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利用去复制、体外和计算机模拟方法快速鉴定小花糖茎中的天然乙酰胆碱酯酶抑制剂

乙酰胆碱酯酶（AChE）抑制是预防阿尔茨海默病（AD）和神经退行性疾病的重要策略。在本研究中，通过比较 UPLC-MS/MS 谱筛选方法和分子对接分析（源自 (Sims) Little 的茎提取物和级分），利用去重复系统快速识别和表征乙酰胆碱酯酶相互作用的化合物。从 的乙酸乙酯提取物中分离并鉴定出 11 种潜在的 AChE 抑制剂，包括一种未描述的生物碱 ()，即甘糖胺 D、八种已知的生物碱 ()、一种类黄酮 () 和一种植物甾醇 ()。评估了这些化合物对AChE的抑制潜力，其中-甲基乙二醇（1,3-dimethoxy-2-羟基-10-甲基-9（10）-吖啶酮（1,3-dimethoxy-2-Hydroxy-10-methyl-9(10)-acridinone）、skimirianine（）和arborine（）被认为是有效的抑制剂，IC 值分别为 39.81 μM、41.53 μM、49.40 μM 和 59.92 μM。值得注意的是，β-甲基乙二醇表现出最高的效力。其中四种有效的 AChE 抑制剂表现出混合型抑制。然而，首先报道了β-甲基乙二醇醚 (β-methylacetolone)、1,3-二甲氧基-2-羟基-10-甲基-9(10)-吖啶酮 (acridinone) () 和 arborine () 调节乙酰胆碱酯酶活性。此外，分子对接显示，与其他化合物相比，-methylacetolone () 具有优异的结合亲和力（–23.749 kcal/mol），主要是通过与 AChE 的外围阴离子位点相互作用，形成氢键，疏水力可能发挥重要作用，与活性空腔中存在 Tyr341、Tyr72、Ser293 和 Arg296 等氨基酸残基，这对于有效和选择性抑制 AChE 活性至关重要。 ADMET 预测表明，arborine ()、skimmianine () 和 -methylacetolone () 表现出良好的穿过血脑屏障的渗透性，而 1,3-dimethoxy-2-Hydroxy-10-methyl-9(10)-acridinone ()表现出相对较低的渗透性。这些发现凸显了这些化合物作为天然乙酰胆碱酯酶抑制剂治疗神经退行性疾病的潜力。