Alprazolam (Alp), a triazolobenzodiazepine, is widely prescribed for the treatment of sleep disorders, anxiety, and panic disorder. While oral administration remains the standard route, its slow onset of action has prompted interest in intranasal delivery as an alternative, offers the potential for direct drug delivery to the brain. This study aims to develop a fast-acting intranasal formulation of Alp (Alp-nd). Safety evaluations were conducted using mucociliary toxicity tests in bullfrogs and ciliary toxicity tests in rats, demonstrating that Alp-nd was safe and did not cause damage to the nasal mucosa. The righting reflex test was employed to assess the dose-response effect of Alp-nd on sleep in mice, revealing a 95 % effective dose (ED