Objectives: The aim of this clinical pilot study was to evaluate the dosage, efficacy, and clinical safety of a single oral dose of pregabalin in cats that experience fear and anxiety when placed into a carrier and transported by car.Methods: Thirteen client-owned cats were enrolled in a blinded, randomized, crossover study with three treatment days approximately 1 week apart. The cats were assigned to receive pregabalin oral solution at dosages of 5 and 10 mg/kg and placebo in a randomized order, one treatment per week. Treatment was administered ~90 min before placing the cat into a carrier and starting transportation. Efficacy was assessed by the owners using a categorical scale and, based on video recordings, by an external observer, both blinded to the treatment.Results: Owners assessed that cats given pregabalin displayed less vocalization, restlessness, and panting during transportation than did cats given placebo. Correlation between owners' and external observer's assessment of the overall treatment effect was good (0.63, p < 0.01), which confirms the owners' ability to observe reliably their own cat's behavior. Transient mild ataxia was the most common adverse event reported. The human commercial formulation used in this study was found difficult or very difficult to administer by 79% of the owners.Conclusions and Relevance: Based on results of this pilot study, a single oral dose of pregabalin was well tolerated and decreased signs of anxiety and fear associated with car transportation in cats, as evaluated by blinded owners and external observer. The use of pregabalin prior to traveling may improve cat welfare and compliance for transportation. Further studies are needed to investigate the use of oral pregabalin in cats to alleviate signs of anxiety and fear associated with transportation and sequelae, like veterinary visits, and to develop a more user-friendly formulation.