Integrated affinity ultrafiltration UHPLC-QE-Orbitrap-MS and molecular docking for efficient screening of aurora kinase a inhibitors from Eomecon chionantha Hance

Aurora kinase A (AURKA) is a crucial serine/threonine protein kinase that plays a pivotal role in regulating core mitotic processes and is widely recognised as a vital therapeutic target for cancer treatment. This study aimed to identify and screen potential AURKA ligands from Eomecon chionantha Hance root extract via a combined approach of ultra-high-performance liquid chromatography-high-resolution mass spectrometry and affinity ultrafiltration mass spectrometry. A total of 134 compounds were identified, including 41 alkaloids. Further screening yielded 10 high-affinity ligands (7 alkaloids and 3 long-chain fatty acyl amides). Molecular docking and molecular dynamics confirmed their stable binding to AURKA. Notably, protopine exhibited the most favourable binding energy, forming multiple hydrogen bonds and hydrophobic interactions with key active site residues. These findings verify the AURKA-binding potential of these natural products, which supply promising candidate ligands and essential references for the research and development of AURKA-targeted inhibitors.Impact Statement Aurora kinase A (AURKA) is a key therapeutic target for cancer, and natural products from medicinal plants are important sources for developing targeted inhibitors. This study screened and identified high-affinity AURKA ligands from Eomecon chionantha Hance root extract, with molecular docking and molecular dynamics verifying their stable binding ability. These natural ligands, especially the alkaloid protopine with favorable binding energy, provide promising candidates for the development of AURKA-targeted cancer inhibitors, and offer valuable references for the research and translation of natural product-based anti-cancer drugs.