Present study provides an in-vitro efficacy of Piper cubeba Linn. f. (Kababchini) in terms of its antibacterial activity against various gram positive and gram negative bacterial strains using Agar well and Kirby Bauer’s Disc Diffusion method according to CLSI guidelines. The efficacy was compared with the standard drugs used as positive control viz. Ciprofloxacin for gram positive strains and Gentamicin as gram negative strains and the solvent used to dissolve the test drug-Dimethyl Sulphoxide (DMSO) as Negative Control. Phytochemical analysis was also done to confirm the presence of various phytoactive constituents present in the test drug and it was revealed that Kababchini contains alkaloids, glycosides, carbohydrates, proteins, tannins, resins and terpenes. It was found to be sensitive towards Staphylococcus aureus, S. epidermidis, Streptococcus mutans, Escherichia coli, Klebseilla pneumoniae, Proteus vulgaris and Pseudomonas aeruginosa. The study demonstrates the in-vitro efficacy of Kababchini to be considered as effective drug target for further screening and its wider use in Unani medicine to combat many infectious diseases effectively.