This study investigates the antinociceptive and antihyperalgesic action caused by i.p., i.t. or i.c.v. injections of amiloride when assessed against formalin, capsaicin-induced licking, acetic acid-induced writhing and glutamate-induced hyperalgesia in mice. The systemic, spinal and supraspinal administration of amiloride causes dose-related antinociception when assessed against acetic acid-induced writhing, formalin and capsaicin-induced licking. In addition, amiloride administered by the same routes produced graded inhibition of glutamate-induced hyperalgesia in mice. Together, these results suggest, that amiloride or its derivatives may constitute a strategy for the development of new antinociceptive drugs.