To explore and compare the prevalence of anxiety and depressive symptoms between first-, second-, and third-year pharmacy students. A repeated-measures study was conducted at two campuses (main and satellite) of the University of Arizona College of Pharmacy. A survey was administered in February 2019 and again in April 2019 during mandatory courses for first-, second-, and third-year Doctor of Pharmacy students to collect seven-item Generalized Anxiety Disorder (GAD-7) scores, nine-item Patient Health Questionnaire (PHQ-9) scores, and demographic information. A chi-square test with a Bonferroni correction was performed to compare the number of students in a class year with clinically significant symptoms, defined as scores of ≥10 for both the GAD-7 and PHQ-9. The survey response rate was 82%. Thirty percent of students self-reported having clinically significant anxiety symptoms and 22% of students self-reported having clinically significant depressive symptoms. More second-year pharmacy students self-reported anxiety and depressive symptoms as the semester progressed. About one in four pharmacy students self-reported clinically significant symptoms of anxiety and depression, and more second-year pharmacy students reported anxiety and depressive symptoms later in the semester. These findings support the need for optimizing the delivery of well-being resources to pharmacy students.