Jiang Haipeng, Xu Wenxi, Bi Tian, Wang Biao, Shi Mengyun, Lv Jiamin, Liu Huwei
School of Chemical Engineering and Pharmacy, Pharmaceutical Research Institute, Wuhan Institute of Technology, Wuhan, 430205, P. R. China.
College of Food Science and Technology, Wuhan Business University, Wuhan, 430056, P. R. China.
Anal Bioanal Chem. 2025 May 30. doi: 10.1007/s00216-025-05911-2.
Screening of ligands from natural products, especially traditional herbs, has always been an important pathway for drug discovery. In the current work, we present a strategy for the screening of active substances from TCM by combination of in silico target prediction, surface plasmon resonance (SPR), and UPLC-HR-MS/MS, using Kansui as an illustration. In the first, in silico target prediction of diterpenoids from Kansui was performed, aiming to improve the success rate of screening. Carbonic anhydrase II (CA II) were within the most likely targets. Then, SPR combined with UPLC-HR-MS/MS analysis was employed for the screening of ligands for CA II from a mixture of Kansui extract and for the structural identification of the captured ingredients. It was found that the ligands of CA II are mainly ingenane-type diterpenoids, which was further validated by enzymatic activity experiments and molecular docking that ingenane-type diterpenoids have more affinity and potency for CA II activation. It is the first time that the ligands of CA II have been screened from the extract of Kansui, and the present ligand screening work also providing clues to the pharmacological mechanisms and rationality of vinegar-processed Kansui.
从天然产物尤其是传统草药中筛选配体一直是药物发现的重要途径。在当前工作中，我们提出了一种通过计算机辅助靶点预测、表面等离子体共振（SPR）和超高效液相色谱-高分辨质谱/质谱联用从中药中筛选活性物质的策略，并以甘遂为例进行说明。首先，对甘遂中的二萜类化合物进行计算机辅助靶点预测，旨在提高筛选成功率。碳酸酐酶II（CA II）是最有可能的靶点之一。然后，采用SPR结合超高效液相色谱-高分辨质谱/质谱分析从甘遂提取物混合物中筛选CA II的配体，并对捕获成分进行结构鉴定。结果发现，CA II的配体主要是瑞香烷型二萜类化合物，酶活性实验和分子对接进一步验证了瑞香烷型二萜类化合物对CA II激活具有更高的亲和力和效力。这是首次从甘遂提取物中筛选出CA II的配体，目前的配体筛选工作也为醋制甘遂的药理机制和合理性提供了线索。
