Alzheimer’s disease (AD) is a chronic and progressive disease of the central nervous system (CNS) with complex underlying pathological mechanisms. The main group of medications used in the AD treatment are acetylcholinesterase inhibitors (donepezil, rivastigmine, galantamine). Despite great advances in neuroscience and new neurotherapeutics, the effectiveness of CNS disease treatment is primarily limited by the existence of a bloodbrain barrier. Thus, brain bioavailability of a systemically administrated drug is reduced. Nasal administration enables direct CNS delivery of the drug. It is characterized by rapid onset of action, non-invasiveness, bypass of the gastrointestinal tract and first-pass metabolism. The aim of this study was to prepare and characterize chitosan/mannitol based donepezil-loaded microspheres and to examine the influence of administration parameters (administration angle in relation to the horizontal plane and inspiratory flow rate) on nasal deposition of prepared microspheres. Donepezil-loaded microspheres met the criteria for nasal application with respect to appropriate entrapment efficiency, drug loading, moisture content, surface charge, and rheological properties. In nasal deposition studies, a 3D printed nasal cavity model, a respiratory pump (for simulating breathing) and a UDSP delivery device were used. Donepezil-loaded microspheres were successfully delivered to the targeted (olfactory) region of the nasal cavity. By reducing the administration angle in relation to the horizontal plane, the proportion of drug delivered to the olfactory region increased. The use of the device at breath-hold resulted in higher drug deposition in the targeted olfactory region, and lower deposition in the anterior and posterior regions, compared to application under breathing conditions.