CD147-SNAP-tag-CMC/HPLC-MS two-dimensional coupling system: a universal platform for screening active components targeting CD147 for anti-liver cancer applications

Established a universal screening platform for CD147-targeted therapeutics. • Integrated the CMC-HPLC system with mass spectrometry (MS) for the identification of unknown compounds. • Two bioactive constituents from Magnolia officinalis, directly interact with CD147. Hepatocellular carcinoma (HCC) is one of the most prevalent digestive malignancies, with CD147 emerging as a promising therapeutic target for HCC. To date, no natural bioactive components targeting CD147 for anti-HCC therapy have been explicitly reported. Given the low toxicity and favorable efficacy of natural small-molecule drugs, identifying and investigating CD147-targeted natural agents is imperative. In this study, based on cell membrane chromatography (CMC) and SNAP-tag protein labeling technology, we constructed a CD147-SNAP-tag-CMC/HPLC-MS two-dimensional coupled system to screen CD147-binding components from Magnolia officinalis , followed by ligand-receptor affinity verification, in vitro pharmacological evaluation, and downstream molecular mechanism validation. Two bioactive compounds, honokiol and magnolol, were successfully identified: both exhibit strong CD147 binding affinity, effectively suppress HCC cell migration, and exert their functions by regulating the CD147/integrin-FAK-paxillin pathway as confirmed by Western blotting. Collectively, the established two-dimensional system serves as a universal platform for CD147-targeted component screening. This study is the first to reveal the strong CD147 affinity of honokiol and magnolol and their anti-migratory effect on HCC cells via modulating the aforementioned pathway.