Kleinia anteuphorbium L. is a medicinal plant widely utilized in traditional medicine by the Souss-Massa population. This study aimed to investigate the best extraction conditions including method and solvents, to achieve the highest polyphenol, flavonoid, antioxidant, and antibacterial activities. The polyphenol and flavonoid contents were quantified using spectrophotometric methods, while the antioxidant activity was assessed through Ferric Reducing Power (FRAP) and 2,2-diphenyl-picrylhydrazyl radical (DPPH) scavenging assays. The antibacterial activity was assessed against four pathogenic clinical isolates :P. aeruginosa, E. coli, K. pneumonia, and S. aureus. The phytochemical analysis revealed that all extracts of Kleinia anteuphorbium exhibited high polyphenol and flavonoid content, along with significant antioxidant activity. Moreover, the extracts showed inhibitory effects against the tested bacteria strains. The GC-MS analysis revealed the existence of abundant bioactive compounds, with potential therapeutic activities. These findings suggest that Kleinia anteuphorbium has promising potential as a medicinal plant.