Alzheimer’s disease (AD) is a multifactorial disorder characterized by exponential loss of memory and cognitive deficit involving several disease modifying targets (amyloid beta, beta-secretase, monoaminoxidase-B, and cholinesterase). The present study explores multi-target directed ligand approach using secondary metabolite reserpine (RES) and ajmalicine (AJM) obtained from Rauwolfia serpentina roots. Novel LCMS and HPLC methods were developed for identification and quantification of reserpine and ajmalicine. In vitro enzyme inhibition assays were performed to evaluate anti-cholinesterase, β-site amyloid cleaving enzyme (BACE-1) inhibition and monoamine oxidase-B (MAO-B) inhibition, further analyzed with in silico analysis. Anti-amyloidogenic potential was studied using anti-aggregation studies along with TEM and circular dichroism (CD) analysis. In vitro neuroprotective potential against Aβ toxicity and anti-oxidative stress was demonstrated using PC12 cell cultures. Reserpine is a more potent dual cholinesterase inhibitor than ajmalicine (IC50 values of 1.7 μM (AChE) and 2.8 μM (BuChE)). The anti-aggregation activity of reserpine (68%) was more than ajmalicine (56%). Both compounds demonstrated neuroprotective activity against Aβ42 (92%) and H2O2 (93%) induced toxicity in PC12 cells against controls. Phytocompounds also inhibited MAO-B and BACE-1 enzymes in concentration dependent manner. Molecular docking studies indicated the strong binding of compounds to the catalytic site of targets. This novel study demonstrated that reserpine and ajmalicine as a multi-target directed ligand that have disease modifying potential for amelioration of AD.

中文翻译：

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Ajmalicine 和利血平：吲哚生物碱作为多靶点定向配体对阿尔茨海默病相关因素的影响

阿尔茨海默病 (AD) 是一种多因素疾病，其特征是记忆力和认知缺陷呈指数级丧失，涉及多种疾病修饰靶点（β-淀粉样蛋白、β-分泌酶、单胺氧化酶-B 和胆碱酯酶）。本研究使用从萝芙木根中获得的次生代谢物利血平 (RES) 和阿马利辛 (AJM) 探索多靶点定向配体方法。开发了新型 LCMS 和 HPLC 方法来鉴定和定量利血平和阿马利辛。进行体外酶抑制试验以评估抗胆碱酯酶、β-位点淀粉样蛋白裂解酶 (BACE-1) 抑制和单胺氧化酶-B (MAO-B) 抑制，并通过计算机分析进一步分析。使用抗聚集研究以及 TEM 和圆二色性 (CD) 分析来研究抗淀粉样蛋白生成的潜力。使用 PC12 细胞培养物证明了针对 Aβ 毒性和抗氧化应激的体外神经保护潜力。Reserpine 是一种比阿马利辛更有效的双重胆碱酯酶抑制剂（IC50 值为 1.7 μM (AChE) 和 2.8 μM (BuChE)）。利血平 (68%) 的抗聚集活性高于阿马利辛 (56%)。两种化合物均表现出针对 Aβ42 (92%) 和 H2O2 (93%) 在 PC12 细胞中对对照诱导的毒性的神经保护活性。植物化合物还以浓度依赖性方式抑制 MAO-B 和 BACE-1 酶。分子对接研究表明化合物与靶标的催化位点有很强的结合力。这项新研究表明，利血平和阿马利辛作为一种多靶点定向配体，具有改善 AD 的疾病修饰潜力。