In this study, we utilized a pharmacological network and bioinformatics approach to investigate the molecular mechanism underlying the resistance of Protopine (PRO) against Triple Negative Breast Cancer (TNBC). To uncover the underlying mechanism of PRO, we employed network pharmacology analysis. We collected and enriched targets using various databases such as TCMSP, SwissTargetPrediction, PubChem, Genecards, and DAVID. Furthermore, we constructed Potential targets network and components-disease-core targets network by STRING 11.5 and Cytoscape 3.7.1 to investigate the association of targets of PRO with disease targets of TNBC. The results of the network pharmacology approach indicated that PRO may play a key role in protein phosphorylation, protein autophosphorylation, Progesterone-mediated oocyte maturation signaling pathway, PI3K-Akt signaling pathway, and acting as targets such as PRKACA, JAK2, CDK2, LRRK2, CCNE1, KDR, JAK1. Our findings suggest that PRO exerts its effects against TNBC through multi-channel and multi-target mechanisms. Therefore, this study provides a basis for further research on the mechanism of action of PRO.