Discovery of drugs rapidly and effectively is an important aspect for Alzheimer's disease (AD). In this study, a novel high-throughput screening (HTS) method aims at screening the small-molecules with amyloid-β (Aβ) binding affinity from natural medicines, based on the combinational use of biolayer interferometry (BLI) and ultra-high-performance liquid chromatography coupled with diode-array detector and quadrupole/time-of-flight tandem mass spectrometry (UHPLC−DAD-Q/TOF-MS/MS) has been firstly developed. Briefly, the components in natural medicines disassociated from biotinylated Aβ were collected to analyze their potential Aβ binding affinity by UHPLC−DAD-Q/TOF-MS/MS. Here, baicalein was confirmed to exhibit the highest binding affinity with Aβ in Scutellaria baicalensis. Moreover, polyporenic acid C (PPAC), dehydrotumulosic acid (DTA), and tumulosic acid (TA) in Kai-Xin-San (KXS) were also identified as potent Aβ inhibitors. Further bioactivity validations indicated that these compounds could inhibit Aβ fibrillation, improve the viability in Aβ-induced PC-12 cells, and decrease the Aβ content and improve the behavioral ability in Caenorhabditis elegans. The molecular docking results confirmed that PPAC, DTA, and TA possessed good binding properties with Aβ. Collectively, the present study has provided a novel and effective HTS method for the identification of natural inhibitors on Aβ fibrillation, which may accelerate the process on anti-AD drugs discovery and development.

快速有效地发现药物是阿尔茨海默病 (AD) 的一个重要方面。在这项研究中，一种新的高通量筛选（HTS）方法旨在从天然药物中筛选出具有淀粉样蛋白-β（Aβ）结合亲和力的小分子，基于生物层干涉仪（BLI）和超高通量的组合使用。 - 首次开发了高效液相色谱与二极管阵列检测器和四极杆/飞行时间串联质谱联用（UHPLC-DAD-Q/TOF-MS/MS）。简而言之，收集与生物素化 A β分离的天然药物中的成分以分析其潜在的 A βUHPLC-DAD-Q/TOF-MS/MS 的结合亲和力。在此，黄芩素被证实与黄芩中的 A β表现出最高的结合亲和力。此外，开心散 (KXS) 中的多孔酸 C (PPAC)、脱氢肿瘤酸 (DTA) 和肿瘤酸 (TA) 也被鉴定为有效的 A β抑制剂。进一步的生物活性验证表明，这些化合物可以抑制 A β纤颤，提高 A β诱导的 PC-12 细胞的活力，降低 A β含量并改善秀丽隐杆线虫的行为能力。分子对接结果证实 PPAC、DTA 和 TA 与 A β具有良好的结合特性。. 总的来说，本研究为鉴定 A β纤颤的天然抑制剂提供了一种新颖有效的 HTS 方法，这可能会加速抗 AD 药物的发现和开发进程。