Addressing antimicrobial resistance (AMR) stands as a major global health challenge threatening humanity. Resolving this issue can be initiated through emphasizing the significance of AMR education among students in health colleges during their undergraduate studies. Hence, the aim of this study is to assess the pharmacy students’ knowledge, attitudes, and practices regarding antibiotic resistance in 7 Middle Eastern countries. A cross-sectional study was conducted among undergraduate pharmacy students at universities in Egypt, Jordan, Saudi Arabia, Lebanon, the United Arab Emirates, Qatar, and Kuwait between March 2021 and January 2022. The first section of the questionnaire gathered demographic information. The knowledge section comprised 7 questions. Subsequently, the questionnaire explored participants’ attitudes (6 items) and practices (2 items) concerning antibiotic resistance. Mann-Whitney and Kruskal–Wallis tests were used to compare the median knowledge score between different demographic groups. Logistic regression was used to estimate odds ratios, with 95% confidence intervals (CIs) for being more knowledgeable about antibiotic resistance. A 2-sided P < .05 was considered statistically significant. A total of 4265 pharmacy students were involved in this study (Egypt (2249), Jordan (n = 704), Saudi Arabia (n = 531), Lebanon (n = 401), United Araba Emirates (n = 130), Qatar (n = 129), and Kuwait (n = 121)). The median knowledge score for the participating pharmacy students was 5.00 (IQR = 4.00–6.00) out of 7, equals to 71.4% with 4th, and 5th year students and bachelor of pharmacy program students have higher odds of being more knowledgeable about antibiotics resistance compared to other students (P < .05). The majority of the students agreed that antibiotic resistance is increasing, they should be more concerned regarding antibiotic consumption and that government should create more awareness of antibiotic resistance, and that they should have enough knowledge to prevent antibiotic resistance. Around 3 quarters of the students (73.0%) confirmed that they take antibiotic only after getting prescription from their physician and almost half (51.7%) reported that they take antibiotic to manage their fever. The study concluded good educational programs in Middle East pharmacy schools with the need for targeted educational interventions promoting responsible antibiotic stewardship practices among future pharmacists.