<p><i>Staphylococcus aureus</i> is a major global health threat due to its increasing antibiotic resistance. The urgent need for alternative treatments has spurred interest in drug repositioning. Drug repositioning the application of approved non-antibiotic agents for new therapeutic uses offers a time-efficient and cost-effective route to identifying novel anti-infective strategies. This study aimed to assess the efficacy of repositioned bioactive agents in treating <i>S. aureus</i> infections by comparing survival outcomes in preclinical animal models. Following the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines, we included <i>in vivo</i> animal models of <i>S. aureus</i> evaluating repositioned non-antibiotic bioactive agents, reporting available survival assay data, with at least one comparator (placebo and/or standard antibiotics). Studies lacking the required data are excluded. Included studies were assessed for risk of bias using the SYRCLE tool. A random effects model was employed to compare bioactive agent effectiveness, with results presented as risk differences and ranked using P-scores. Ultimately, 17 eligible studies were selected from 8594 screened records. The majority of the studies (88.2%) used mice, with intraperitoneal infection models (41.1%) and anticancer agent categories (23.5%) being the most common. Several repositioned drugs showed significantly better survival rates compared to the negative control. Moreover, none of the treatments significantly outperformed the positive controls. Our analysis found that, compared to the negative and positive controls, oral FdUrd and isopropoxy benzene guanidine had the highest P-scores (0.98 and 0.9, respectively). These findings support the continued exploration of non-antibiotic repurposing strategies as viable adjuncts or alternatives in managing drug-resistant <i>S. aureus</i> infections, with implications for translational and formulation development in antimicrobial drug delivery.</p>