In this study, an efficient strategy developed by integrating UPLC-Q/TOF-MS, network pharmacology, and molecular simulation, was proposed and applied for rapidly screening bioactive candidates from ginger. A UPLC-Q/TOF MS-guided isolation targeting non-volatile pungent compounds resulted in the isolation and identification of 19 compounds in the rhizome of Zingiber officinale, including six new compounds (1–6). Based on target prediction and Gene Ontology (GO), the primary biological function of compounds was predicted to be associated with cancer and the key target was VEGFR2 (vascular endothelial growth factor receptor 2). Moreover, cytotoxic activity assays demonstrated that the isolated compounds had potential anti-proliferative effects on MDA-MB-231, A549 and HCT116 cells. In particular, compounds 7 and 8 exhibited the highest cytotoxicity against HCT116 compared with the other cell lines, with IC₅₀ values ranging from 4.70 to 7.40 μM. In addition, VEGFR2 inhibition of compounds 7 and 8 was validated based on enzyme activity assays and their interaction mechanisms were illuminated through molecular simulations. These experimental data are consistent with the calculated results, indicating the veracity of the proposed method. In conclusion, the integrated strategy is a quick and efficient way to explore bioactive compounds as well as research the possible targets, providing us with a good possibility of screening new lead compounds from natural sources.

中文翻译：

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从姜中分离出来的非挥发性辛辣化合物及其作用机理†

在这项研究中，提出了一种通过整合UPLC-Q / TOF-MS，网络药理学和分子模拟而开发的有效策略，并将其用于快速筛选生姜中的生物活性候选物。以UPLC-Q / TOF MS为导向的针对非挥发性刺激性化合物的分离，导致了姜黄根茎中19种化合物的分离和鉴定，包括6种新化合物（1–6）。根据靶标预测和基因本体论（GO），可以预测化合物的主要生物学功能与癌症有关，关键靶标是VEGFR2（血管内皮生长因子受体2）。此外，细胞毒性活性测定表明，分离的化合物对MDA-MB-231，A549和HCT116细胞具有潜在的抗增殖作用。特别地，与其他细胞系相比，化合物7和8对HCT116表现出最高的细胞毒性，IC ₅₀值为4.70至7.40μM。此外，VEGFR2对化合物7和8的抑制作用基于酶活性测定法进行了验证，并通过分子模拟阐明了它们的相互作用机理。这些实验数据与计算结果一致，表明了所提方法的准确性。总之，综合策略是探索生物活性化合物以及研究可能目标的一种快速有效的方法，为我们提供了从天然来源中筛选新的先导化合物的良好可能性。