齐留顿
化学
色谱法
超滤(肾)
质谱法
液相色谱-质谱法
格列吡嗪
花生四烯酸5-脂氧合酶
酶
生物化学
医学
内分泌学
糖尿病
花生四烯酸
作者
Siqi Wang,Jiaxin Huai,Ying Shang,Linlin Xie,Xiaotong Cao,Jun Liao,Teng Zhang,Ronghua Dai
标识
DOI:10.1016/j.jep.2020.112733
摘要
Zi-shen pill (ZSP), a traditional Chinese medicine, is widely used for the treatment of benign prostatic hyperplasia (BPH) and has remarkable curative effect. To screen the potential 5-Lipoxygenase(5-LOX) inhibitors from ZSP extract. A new approach based on affinity ultrafiltration-ultra performance liquid chromatography-mass spectrometry(UPLC-MS) was established and validated. Zileuton and glipizide were chosen as positive and negative control drug, respectively. For better screening result, the concentration of 5-LOX enzyme, incubation temperature and time, pH and ion strength were optimized. In addition, 5-LOX inhibitory assay in vitro and molecular docking technique were used for further verification. 20 compounds were characterized in the ultrafiltrate by high resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and 16 ligands showed binding ability to 5-LOX. Among them, six ligands were deduced as high-potential 5-LOX inhibitors with their high specific binding values (>2.0). The inhibitory activities of anemarrhenasaponin I, timosaponin AI, nyasol and demethyleneberberine were confirmed by the 5-LOX inhibitory assay for validating the reliability of affinity ultrafiltration approach and the computer-simulated molecular docking technique further clarified the possible mechanism of action between the active compounds and the 5-LOX active sites.
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