Aurone Derivatives as Inhibitors of ERK Phosphorylation in HCC827 Non-Small Cell Lung Cancer Cells.

INTRODUCTION Non-Small Cell Lung Cancer (NSCLC) continues to be a primary contributor to cancer- related death, with limited long-term efficacy of current therapies due to resistance and tumor heterogeneity. Focusing on downstream signaling pathways, such as the EGFR-mediated MAPK/ERK pathway, represents a promising therapeutic strategy. In this study, we designed and synthesized twenty hydroxy- and methoxysubstituted (Z)-2-benzylidenebenzofuran-3(2H)-one derivatives and evaluated their anticancer potential in EGFR-mutant HCC827 NSCLC cells.

Methods

Twenty (Z)-2-benzylidenebenzofuran-3(2H)-one derivatives synthesized in this study were designed and identified based on NMR spectroscopy and high-resolution mass spectroscopy. ERK1/2 Mitogen-Activated Protein Kinase (MAPK) inhibition assay was performed, and one of them, d4, (Z)-4-methoxy-2-(2,3,4- trimethoxobenzylidene)benzofuran-3(2H)-one, was selected for Density Functional Theory (DFT) calculations and in silico docking with ERK1.

Results

Immunoblot analysis revealed that most compounds effectively inhibited ERK1/2 phosphorylation, with several derivatives showing significant suppression of MAPK signaling. Among them, derivative d4 exhibited the most potent biological activity, including strong inhibition of ERK activation, reduced cell viability, and marked suppression of colony formation. DFT calculations indicated that d4 exhibits favorable thermodynamic stability and higher reactivity than inactive compounds, as evidenced by a smaller HOMO-LUMO gap. Molecular docking analysis further demonstrated that d4 binds stably within the ERK1 active site, exhibiting interactions comparable to those of a known ligand.

Discussion

Biological and computational evaluations highlight derivative d4 as a highly effective smallmolecule inhibitor of ERK phosphorylation in EGFR-mutant HCC827 NSCLC cells. Molecular docking revealed that d4 occupies the ERK binding pocket, forming interactions comparable to those of established ligands, resulting in robust anti-clonogenic phenotypes. Overall, our findings highlight the therapeutic potential of the (Z)-2-benzylidenebenzofuran-3(2H)-one structure in inhibiting hyperactivated MAPK signaling. This study provides a validated chemical platform for subsequent structural optimization and supports the continued development of aurone-based scaffolds as anticancer therapeutics.

Conclusion

Collectively, these findings identify d4 as a promising lead compound that targets ERK-mediated signaling in HCC827 NSCLC cells.