We aimed to investigate antinociceptive activity and median lethal dose (LD50) of water extract of Ballota glandulosissima Hub.-Mor&Patzak in mice. In this study, water extract of Ballota glandulosissima was investigated for antinociceptive activity using acetic acid-induced writhing and tail-flick tests. Acetyl salicylic acid and morphine were used as the reference drug. Mice were injected Ballota glandulosissima extract intraperitoneally in doses of 100 mg/kg and 200 mg/kg, respectively. The extract of Ballota glandulosissima caused dose related inhibition in the acetic acid-induced ab dominal stretching response in mice. The extract of Ballota glandulosissima also showed significant changes in the nociceptive threshold of the tail-flick test. The motor coordination of mice treated with the water extract was evaluated by using the rotarod test and was found to be not impaired in comparison with that of control mice.The results obtained in the present study indicate that the water extract of Ballota glandulosissima has promising antinociceptive activity. The LD50 of Ballota glandulosissima was determined as 8.885 g/kg.Keywords: Ballota glandulosissima, lamiaceae, antinociception, writhing test, tail-flick test, rotarod test.