State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Bioactive Natural Product Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, PR China.
State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Bioactive Natural Product Research, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, PR China.
J Chromatogr A. 2020 Aug 16;1625:461294. doi: 10.1016/j.chroma.2020.461294. Epub 2020 Jun 3.
It is significant to precisely isolate potential active compounds from medicinal herbs containing multiple compounds. Herein, a new strategy for precise separation of lysine-specific demethylase 1 (LSD1) inhibitors from the rhizome of Corydalis yanhusuo (RCY) using counter-current chromatography (CCC) guided by molecular docking and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis was established. First, representative alkaloids from RCY were docked with LSD1 for screening active skeleton compounds. Simultaneously, the crude extract of RCY was preliminarily separated via pH-zone refining CCC. Subsequently, guided by LC-MS/MS analysis of the fragmentation pathways, three potential active fractions were obtained, followed by further online-storage and recycling CCC separation. Finally, three high-purity target quaternary alkaloids compound 3 (dehydrocorydaline), 7 (coptisine), and 8 (columbamine) were successfully isolated as a new class of potential natural LSD1 inhibitors by only one CCC instrument with multiple modes. Compound 3, with the highest LSD1 inhibition ratio of 2.44 μM, was tested for its ability to inhibit tumor invasion and metastasis in U2OS cells. Therefore, the CCC separation guided by virtual screening is a promising method for the targeted isolation of enzyme inhibitors from medicinal herbs.
从含有多种化合物的药用植物中精确分离潜在的活性化合物非常重要。本文建立了一种新的策略，即通过分子对接和液相色谱-质谱/质谱（LC-MS/MS）分析指导的逆流色谱（CCC），从延胡索（RCY）的根茎中精确分离赖氨酸特异性去甲基酶 1（LSD1）抑制剂。首先，用 LSD1 对接 RCY 中的代表性生物碱，筛选活性骨架化合物。同时，用 pH 区带精制 CCC 对 RCY 的粗提取物进行初步分离。然后，根据 LC-MS/MS 分析的碎片途径，得到三个潜在的活性馏分，然后通过在线储存和循环 CCC 分离进一步分离。最后，仅使用一台具有多种模式的 CCC 仪器，成功地从三种高纯度目标季铵生物碱化合物 3（脱氢紫堇碱）、7（小檗碱）和 8（苦参碱）中分离出作为一类新的潜在天然 LSD1 抑制剂。化合物 3 的 LSD1 抑制率最高，为 2.44 μM，用于测试其在 U2OS 细胞中抑制肿瘤侵袭和转移的能力。因此，虚拟筛选指导的 CCC 分离是从草药中靶向分离酶抑制剂的一种很有前途的方法。
