S. aureus isolate was inoculated and cultured to exponential growth phase (OD600 at 0.5) in TSB and divided into the control group and two drug treated groups with different concentrations. Pinaverium bromide was added to the drug treated group until a final concentration of 1/2× MIC and 1/4× MIC reached, and DMSO added in the control group. Then, cultures in all groups were cultivated for 2h at 200 rpm. The microbial was centrifuged at 12000 rpm for 10 min, washed twice with Phosphate Buffer Saline (PBS). Lyse the cells by bead-beating at 4 °C in RIPA lysis buffer (1% Triton X-100, 1% deoxycholate, 0.1% SDS), The protein reduction, alkylation and tryptic digestion were followed as filter-aided sample preparation (FASP). The digestion products were analyzed with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), followed by label-free quantitative analysis. The MS experiment was performed on a Q Exactive plus mass spectrometer.