Untargeted UHPLC-Q Exactive Orbitrap HRMS-Based Network Pharmacology for the Discovery of Anti-Breast Cancer Compounds from Spatholobus littoralis Stems Fractions and Experimental Validation

Spatholobus littoralis has been traditionally used by the Dayak tribe as a medicine to treat breast cancer. However, its bioactive components and mechanisms of action remain unclear. To identify the bioactive compounds in the aqueous fraction of the 70% ethanol extract of S. littoralis stems and elucidate their mechanisms of action as an anti-breast cancer agent. The fractions of the 70% ethanol extract were tested for antioxidant activity and aqueous fraction was analysed for bioactive compounds by using UHPLC-Q-Orbitrap-HRMS, network pharmacology, molecular docking, and experimental validation. The protein-protein interaction (PPI) analysis identified 10 target genes related to breast cancer, with KEGG pathway analysis indicating activity in breast cancer. The molecular docking analysis showed binding affinities from −5.7 to −7.5 kcal/mol. The experimental validation using cytotoxicity testing resulted in an IC50 value of 121.66 µg/mL. Several bioactive compounds from the aqueous fraction exhibited anti-breast cancer activity via EGFR tyrosine kinase inhibition.