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Background: Non-adherence is a major concern in the treatment of diabetes mellitus and undermines the goals of treatment. The objective of this study was to determine the magnitude of non-adherence and its contributing factors among diabetes mellitus patients attending the diabetes mellitus clinic at Mbarara Regional Referral Hospital. Objective: To assess prevalence and factors contributing to non-adherence to antidiabetic medication among diabetes mellitus patients in the diabetic clinic at Mbarara Regional Referral Hospital. Methods: A descriptive cross-sectional study was adopted at the diabetes clinic, Mbarara Regional Referral Hospital, between July and October 2020. Study participants were systemically sampled, and data regarding their medication non-adherence was collected using a structured questionnaire, based on the Hill-Bone medication adherence scale. Data entry was done using Microsoft Excel Version 2010, and analysis was carried out using STATA version 13. The odds ratio was used to determine the strength of association between diabetic medication non-adherence and associated factors. The cutoff value for all statistical significance tests was set at p < 0.05 with a confidence interval of 95%. Results: A total of 257 participants were recruited with a 100% response rate. More than one-third (98, 38.1%) of the participants were non-adherent to their antidiabetic medication. Age above 60 years (AOR = 6.26, 95% CI = 1.009– 39.241, P = 0.049) and duration of diabetes mellitus above 5 years (AOR = 1.87, 95% CI = 1.034– 3.392, P = 0.038) were independently associated with non-adherence to antidiabetic medication. Conclusion: The prevalence of non-adherence to antidiabetic medication was higher than that revealed in previous studies in Uganda. Patients with age above 60 years were six times more likely to be non-adherent to their anti-diabetic medications. Patient education is important to address the challenges of medication non-adherence. [ABSTRACT FROM AUTHOR]