In summary, the current study highlights the influence of both celastrol and terbinafine on S. aureus staphyloxanthin, antibiotic susceptibility and host pathogenesis. Treating of S. aureus infections is problematic, as S. aureus produces numerous virulence factors that shield bacteria from external environment, antibiotics and host immune response. Importantly, staphyloxanthin biosynthesis in S. aureus was markedly reduced upon treatment with either celastrol or terbinafine. The inhibitory potential of celastrol and terbinafine on both staphyloxanthin and bacterial pathogenesis was fully investigated herein. Moreover, celastrol and terbinafine synergized the activity of commonly used antibiotics such as ampicillin, cefotaxime, gentamicin, azithromycin, and ciprofloxacin against S. aureus. Importantly, celastrol and terbinafine exhibited a significant inhibitory effect on S. aureus pathogenesis which was extensively characterized in vivo using mice infection model. To our knowledge, current study is the first one that characterized the effect of both celastrol and terbinafine on staphyloxanthin production and S. aureus virulence potenial. Altogether, current findings clearly suggest the involvement of celastrol and terbinafine in treatment strategies against S. aureus. Celastrol and terbinafine could be used in combination with commonly used antibiotics against S. aureus in order to control infections caused by this important pathogen.