Purpose <p>Pramlintide is an amylin analog developed as a complementary treatment for diabetes. However, it requires several subcutaneous injections, reducing patients’ adherence. Since the intranasal route might be an alternative for drug administration, we evaluated whether intranasal pramlintide treatment exerts comparable actions with intraperitoneal administration.</p> Methods <p>Adult male Swiss mice were submitted to a refeeding test in a dose-response study with intraperitoneal (PRAM i.p.) or intranasal (PRAM i.n.) pramlintide administration. Intraperitoneal liraglutide served as a positive control (LIRA). Then, the selected dose was administered to analyze gastric emptying after an acute exposure. We also evaluated an 8-day treatment (once daily) to determine food intake and body mass. Blood glucose and plasma triacylglycerides were measured on the euthanasia day.</p> Results <p>In the refeeding test, the anorexigenic dose for the PRAM i.p. or LIRA i.p groups was 200 µg/kg and 400 µg/kg, respectively. The PRAM i.n. group (200 µg/kg) exhibited a trend for that. The reduction in gastric emptying occurred for all treated groups compared with their respective controls (vehicle-treated). Neither the PRAM i.p. nor the PRAM i.n. groups exhibited reduced body mass and food intake in the subchronic experiment. No impact on biochemical parameters was observed regardless of the route of pramlintide administration.</p> Conclusion <p>Although intranasal pramlintide is not comparable in magnitude to intraperitoneal administration at an equivalent administered dose, our evidence corroborates the development of novel intranasal formulations destined to overpass the bioavailability issue and potentially serve as an alternative route.</p> Graphical Abstract <p></p>