Abstract The aim of this study was to investigate the phenolic compound from Pinaropappus roseus , and its human breast anticancer properties. The phenolic molecules were isolated from the aqua-ethanoic extract isolated from the leaves of P. roseus by solid-phase extraction (SPE). The total phenolic content was determined by the Folin-Ciocalteu technique. The profile of phenolic compounds was analyzed by mass spectrometry (LC-ESI-MS 2 ). The quantification of phenolic molecules identified by mass spectrometry was carried out by UV-spectrophotometry (LC-PDA). The in vitro cytotoxicity assay was carried on MCF-7 and HMEC cell lines using the MTT assay method. The docking simulation was carried out in anti-apoptotic proteins, Bcl-2 and Bcl-xL. Six phenolic compounds were identified of which the apigenin (37.5 mg CAE g db −1 ) was the most abundant compound. In the in vitro anti-cancer assay, the IC50 for the MCF-7 cells was of 426.15 µg g db −1 at 24h and 297.40 µg g db −1 at 48 h for the maximum evaluated phenolic extract concentration. The rutin and the chlorogenic acid showed the higher binding energies in the docking simulation for the active sites of the Bcl-2 and Bcl-xL proteins respectively. The phenolic compounds of P. roseus have cytotoxic activity against human breast cancer (MCF-7 cell line) and a low cytotoxic activity against normal human epithelial cells (HMEC cell line).These results suggest that the phenolic extract of P. roseus may have therapeutic potential against human cancer pathologies.