Piper nigrum finds an extensive application in antibacterial preparations belonging to Ayurvedic system of medicine. A bioguided extraction and fractionation of the petroleum ether extract of the berries of P. nigrum afforded 2E, 4E, 8Z-N-isobutyleicosatrienamide (1), pellitorine (2), trachyone (3), pergumidiene (4) and isopiperolein B (5). Pergumidiene and trachyone are isolated for the first time from P. nigrum. All the isolated compounds were active against Bacillus subtilis, Bacillus sphaericus, and Staphylococcus aureus amongst Gram + ve bacteria, and Klebsiella aerogenes and Chromobacterium violaceum among Gram -ve bacterial strains.