The in vitro study on TNBC cell lines demonstrated that the candidate compound F25 exhibits potent anti-tumor metabolic activity in MDA-MB-231 cells. Specifically, F25 significantly reduced the extracellular acidification rate (ECAR), indicating its potential to disrupt tumor cell metabolism by targeting the lactate transporter MCT4 pathway. Furthermore, F25 treatment, either alone or in combination with cisplatin (CDDP), led to a marked reduction in intracellular pH, with the combination treatment showing enhanced efficacy. Protein expression analysis revealed elevated MCT4 and reduced MCT1 expression in MDA-MB-231 cells, suggesting a mechanistic link to F25’s mode of action. These findings highlight the therapeutic potential of F25 in targeting tumor metabolism.