The antiopiate activities of Tyr-MIF-1 (Tyr-Pro-Leu-Gly-NH2) and some of its representative analogs were tested in two animal models of antinociception. Doses of the tetrapeptides as low as 0.001 mg/kg injected peripherally could block the analgesic effects of morphine in both the tail-flick test of mild thermal pain induced by heat and the scratching test of mild chemical pain induced by hypertonic saline. These tetrapeptides showed cross-reactivity in the radio-immunoassay (RIA) used to identify the presence of Tyr-MIF-1 in brain extracts and in the brain membrane binding assay. Only Tyr-MIF-1, however, eluted at the position of the immunoreactive peak after gel filtration chromatography and high performance liquid chromatography (HPLC). The results support the concept that peptides with anti-opiate activity can exist in the brain.