The current surge of multidrug resistant infections has led to many cases of treatment failure in clinical practices. This has necessitated the search for new drugs from natural sources which could be used to curb this menace. Paullinia pinnata (Linn.) leaves are deployed traditionally for the treatment of various ailments such as malaria and the treatment of wounds. The essence of this study is to assess the antibacterial potential of P. pinnata leaf fractions on four antibiotic-resistant bacteria isolated from treatment failed wounds. The n-hexane and ethyl acetate fractions were obtained from the methanol extract of the leaves of P. pinnata by the one-solvent system fractionation. The antibacterial capacity of the fractions was determined by the agar well diffusion method and gentamicin (30 µg/mL) was the control. Clinical isolates of two Gram-positive (Staphylococcus aureus and Bacillus cereus) and two Gram-negative (Klebsiella pneumoniae and Proteus vulgaris) bacteria were used. The fractions showed no activity to moderate activity at lower concentrations. The ethyl acetate fraction showed bacteriostatic activity in a dose-dependent manner and was better than that of n-hexane, and could therefore be a good drug target for antimicrobial therapies, especially for wound treatment.