High-expression EGFR/cell membrane chromatography-online-high-performance liquid chromatography/mass spectrometry: rapid screening of EGFR antagonists from Semen Strychni.

RATIONALE

Traditional methods for screening active compounds from complex system such as traditional Chinese medicines are relatively cumbersome and time-consuming. In order to improve this situation, we established an online analytical method for screening, separation and identification EGFR antagonists from traditional Chinese medicines, which is described in this study.

METHODS

Cells with high EGFR expression levels were used to prepare the cell membrane stationary phase for the EGFR cell membrane chromatography model with the purpose of screening active compounds. Separation of the retention fractions was achieved by the high-performance liquid chromatography, and identification was conducted via electrospray ionization quadrupole mass spectrometry. The inhibitory effects of active compounds on EGFR cell growth were also demonstrated in vitro.

RESULTS

The screening results showed that vauquline and strychnine from Semen Strychni could be active components acting on EGFR similarly to gefitinib as a control drug. Results from biological trials showed that vauquline and strychnine inhibited cell proliferation of HEK293/EGFR cells, and inhibited Erk phosphorylation, and can effectively reduce expression of downstream signaling molecules.

CONCLUSIONS

This EGFR cell membrane chromatography-online-high-performance liquid chromatography/tandem mass spectrometry method can be applied for rapid screening, separation and identification of EGFR antagonists from traditional Chinese medicines and should be useful for drug discovery with natural medicinal herbs.