Screening of 5‐Lipoxygenase Inhibitors From Ziziphi Spinosae Semen (seed of Ziziphus jujube) Using an Online Activity Detection System Combined With Molecular Docking

• DOI: 10.1002/sscp.70026
• Publication Date: 2025-04-01T19:38:33Z
• Authors: Yuangui Yang; Yaya Zhu; Hongbo Xu

ABSTRACTTo develop and refine an online activity detection system for in vitro screening of bioactivity and structure–activity relationship of Ziziphi Spinosae Semen (ZSS.). An online active detection system integrating ultrahigh performance liquid chromatography, photodiode array detection, mass spectrometry, and 5‐lipoxygenase fluorescence detection was established. Additionally, molecular docking were conducted to predict interactions between the potential active ingredients and 5‐lipoxygenase, facilitating the swift identification of antiinflammatory agents from the ZSS. The optimal conditions for the reaction system were identified as a temperature of 30°C, a flow rate of 0.2 mL/min, and a 0.1 mol/L phosphate‐buffered saline solution at pH 8.1, with an enzyme concentration of 1.5 × 104 U/mL. Using this system, 10 constituents from the ZSS. exhibited potential binding affinity with 5‐lipoxygenase. Subsequent molecular docking of these 10 candidate bioactive molecules with 5‐lipoxygenase indicated that rutin, zivulgarin, sanjoine, 6‴‐feruloylspinosin, 6‴‐(3‴′,4‴′,5‴′‐trimethoxyl)‐(E)‐cinnamoylspinosin, nummularine B, vicenin II, 6‴‐O‐(3R‐1‐N‐β‐d‐glucopyranosyl‐2‐oxo‐3‐hydroxy‐indole‐3‐acetyl)spinosin, and apigenin displayed significant binding capabilities with 5‐lipoxygenase. This system offers a robust platform for screening 5‐lipoxygenase inhibitors, supporting the identification of premium ZSS. This methodology presents a novel approach for the rapid identification of potential antiinflammatory bioactive components in traditional Chinese medicine.