在我们寻找从Clinacanthusnutans(Burm.f.)Lindau叶发现晚期糖基化终产物(AGEs)抑制剂的过程中,我们进行了生物活性分子网络。该方法整合了LC-MS2概况和体外抗糖基化数据以预测生物活性化合物。我们开始筛选三种提取物:100%乙醇,70%乙醇和100%水的体外抗氧化活性,总酚含量(TPC)和沙己糖苷含量。在这些提取物中,100%乙醇提取物表现出最高的总AGEs抑制作用(IC50=80.18±11.6μg/mL),DPPH清除活性(IC50=747.40±10.30μg/mL)和总酚含量(26.54±2.09μgGAE/mg提取物)。有趣的是,100%乙醇提取物含有最低量的沙夫糖苷,表明其他植物化学物质参与了抗糖基化作用。在来自100%乙醇提取物的级分中检测到的401LC-MS特征的分子网络和计算机结构注释预测了21种生物活性化合物(p<0.05,r>0.90),包括几种C40类胡萝卜素,含有四吡咯结构和脂肪酸的生物碱。相反,所有酚类物质与抗糖基化作用的相关性均较弱.这些预测在体外进一步验证,其中类胡萝卜素叶黄素显示一半最大抑制浓度,IC50=96±8μM,选择的类黄酮-C-糖苷表现出较弱的抑制作用(IC50在568和1922μM之间)。值得注意的是,冻干叶片的叶黄素含量(12.42±0.82mg/100g)高于烘干叶片,尽管前者与汞含量升高有关。总之,坚果梭菌表现出潜在的抗糖基化和抗氧化活性,叶黄素被确定为主要的生物活性成分。
In our quest to discover advanced glycation end products (AGEs) inhibitors from Clinacanthus nutans (Burm.f.) Lindau leaves, we conducted a bioactivity-based molecular networking. This approach integrates LC-MS2 profiling and in vitro
antiglycation data to predict bioactive compounds. We began by screening three extracts: 100% ethanol, 70% ethanol and 100% water alongside the in vitro antioxidant activity, total phenolics content (TPC) and schaftoside content. Among these extracts, 100% ethanol extract exhibited the highest total AGEs inhibition effects (IC50 = 80.18 ± 11.6 μg/mL), DPPH scavenging activity (IC50 = 747.40 ± 10.30 μg/mL) and TPC (26.54 ± 2.09 μg GAE /mg extract). Intriguingly, 100% ethanol extract contained the lowest amount of schaftoside, suggesting the involvement of other phytochemicals in the
antiglycation effects. The molecular networking and in silico structural annotations of 401 LC-MS features detected in the fractions from 100% ethanol extract predicted 21 bioactive compounds (p < 0.05, r > 0.90), including several C40 carotenoids, alkaloids containing tetrapyrrole structures and fatty acids. On the contrary, all phenolics showed weak correlations with
antiglycation effects. These predictions were further validated in vitro, where carotenoid lutein showed half maximal inhibitory concentration, IC50 = 96 ± 8 μM and selected flavonoid-C-glycosides exhibited weaker inhibitions (IC50 between 568 and 1922 μM). Notably, lutein content was higher in freeze-dried leaves (12.42 ± 0.82 mg/100 g) than oven-dried, although the former was associated with elevated mercury levels. In summary, C. nutans exhibited potential
antiglycation and antioxidant activity, and lutein was identified as the main bioactive principle.