为明确葛根中能抑制黄嘌呤氧化酶(xanthine oxidase,XOD)活性的化合物,该研究首先制备了固定化XOD的核壳型磁性二氧化硅(Fe3O4@SiO2-XOD),纳米颗粒。结合高效液相色谱-质谱联用技术与分子对接模拟,对葛根提取液中经筛选获得的化合物进行分离,并分析其与XOD的结合特性。结果表明,所制备的Fe3O4@SiO2-XOD纳米颗粒对XOD结合物表现出良好的筛选特性,能够有效应用于配体筛选实验。从葛根提取液中成功筛选鉴定出15种化合物,包括异黄酮类9种、酚酸类3种、植物甾醇类2种以及脂肪酸类1种。进一步对黄嘌呤、次黄嘌呤、别嘌呤醇以及筛选所得化合物与XOD进行分子对接模拟,发现15种化合物与XOD的结合能均优于底物黄嘌呤、次黄嘌呤以及别嘌呤醇,其中异黄酮类整体表现优异,位列第一梯队;植物甾醇依赖疏水作用位列第二梯队;酚酸类及脂肪酸类均位列第三梯队,15种化合物均为有效XOD抑制剂。该研究建立了一种从天然复杂混合物中快速、靶向鉴定生物活性化合物的有效方法。
To identify compounds derived from kudzu root (Pueraria lobata) with inhibitory activity against xanthine oxidase (XOD), this study first prepared core-shell magnetic silica nanoparticles immobilized with XOD (Fe3O4@SiO2-XOD).By integrating high-performance liquid chromatography mass spectrometry and molecular docking simulations, compounds isolated from the screened kudzu root extract were further separated, and their binding characteristics with XOD were analyzed.Results indicated that the synthesized Fe3O4@SiO2 XOD nanoparticles exhibited excellent specificity for XOD-binding ligands and proved highly effective for ligand screening applications.A total of 15 compounds were successfully identified from the kudzu root extract, including 9 isoflavones, 3 phenolic acids, 2 phytosterols, and 1 fatty acid.Molecular docking simulations were performed involving xanthine, hypoxanthine, allopurinol, and the screened compounds against XOD.Results demonstrated that all 15 compounds exhibited stronger binding affinity (lower binding energy) to XOD compared to the substrates (xanthine and hypoxanthine) and the reference inhibitor allopurinol.Among these, isoflavones showed the highest potency, forming the first tier of inhibitors.Phytosterols, which primarily rely on hydrophobic interactions, constituted the second tier.Phenolic acids and fatty acids comprised the third tier.All 15 compounds were identified as effective XOD inhibitors.This study establishes an efficient and targeted methodology for the rapid identification of bioactive compounds from complex natural mixtures.
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