Introduction Lesbian, gay, bisexual, transgender, and queer (LGBTQ+) individuals continue to show disproportionate prevalence of health disparities despite an increase in visibility in recent years. Educational endeavors in professional programs have been proven to increase awareness of culturally inclusive care. Objectives This study assessed self‐reported knowledge and motivation for change of pharmacy students toward providing affirming care for LGBTQ+ patients after attending LGBTQ+ cultural sensitivity training. The second objective of this study is to assess whether first‐ (P1) or second‐year (P2) students would gain greater benefit from the training from their survey responses. Methods Doctor of Pharmacy (Pharm.D.) students at a large metropolitan area in California were provided LGBTQ+ cultural sensitivity training as part of a co‐curricular initiative. Anonymous surveys were administered via pre‐ and post‐training to assess perception of knowledge gained and motivation to adopt LGBTQ+ inclusive practices. Perception and motivation questions were graded on a Likert Scale with 1 being no knowledge and 5 being extensive knowledge, or 1 being strongly disagree and 5 being strongly agree. Results of surveys before and after the training were analyzed with Wilcoxon Signed Rank Sum Test. Chi‐square analysis was conducted to compare survey results for P1 and P2 students. Results Of the 168 students who completed the competency training and both surveys, 70% were P1 students and 30% were P2 students. After the cultural competency training, students reported a statistically significant improvement in their knowledge and motivation to provide affirming care for LGBTQ+ patients (P < 0.0001). Conclusion The two‐hour co‐curricular LGBTQ+ cultural competency training demonstrated improvements in the knowledge and attitude of students toward providing inclusive and competent care to gender and sexual minorities. LGBTQ+ related content should be integrated into Pharm.D. education early in the curriculum to address several objectives in cultural sensitivity.