Metabolomics‐Driven Bioactivity‐Guided Isolation and Molecular Docking Reveal Antibreast Cancer Metabolome Heterogeneity in Polygonum spp

ABSTRACT The rise of triple‐negative breast cancer (TNBC) warrants the urgent need to identify novel therapies. Herein, untargeted metabolomics‐guided isolation revealed the metabolome fingerprints of Polygonum bistorta (PB) and Polygonum viviparum (PV) in correlation with their TNBC inhibitory activity. Phytochemical analysis annotated 209 metabolites, with flavonoids (45 compounds) as the major class, followed by phenolic acids (25), tannins (24), and hydroxycinnamic acids (18). Multivariate data analysis identified chlorogenic acid, 6‐ O ‐galloylarbutin, and procyanidin B1 as key markers of PV, whereas naringenin‐ O ‐glucoside, catechin, and crataegunin D characterized PB. Among different polar extracts, ethyl acetate extract of PB (PB‐EA) and the n ‐butanol extract of PV (PV‐NB) exhibited the strongest cytotoxicity against 4T1 cells, with IC 50 values of 23.28 ± 0.90 and 39.11 ± 2.36 μg/mL, respectively. Subsequently, the same extracts significantly suppressed tumor growth in the 4T1‐Luc orthotopic xenograft mouse model compared with their respective model groups ( p < 0.001, p < 0.01). Bioassay‐guided isolation yielded 8 compounds, including crataegunin D, which isolated from Polygonaceae plants for the first time, displaying IC 50 values of 24.91 ± 2.43 and 20.65 ± 1.77 μM against 4T1 and MDA‐MB‐231 cells, respectively. Molecular docking revealed binding free energies (ΔG) ranging from −52 to −48 kcal/mol targeting EGFR and CDK receptors.