Introduction Chemotherapies are handled using Good Manufacturing Practices, which ensure asepsis and high-quality production. Continuous education is compulsory and usually includes theoretical and practical exercises. Objectives This work aimed to validate an innovative method of teaching good manufacturing practices based on an escape room mixing simulation and gaming. Method Pairs of learners were locked in a simulated clean room (Esclean Room) and had 1 hour to produce a chemotherapy and escape by finding solutions to 23 “Good Manufacturing Practices mysteries” linked to combination locks. To measure the experiment’s impact on teaching, questionnaires including the 23 mysteries (in different orders) were filled in before, just after and one month after escape from the Esclean Room. Pharmacy staff’ degrees of certainty were noted for each question. A satisfaction survey was completed. Results Seventy-two learners (29% senior pharmacists, 14% junior pharmacists, and 57% pharmacy technicians) escaped the Esclean Room and 56 answered every questionnaire. The educational intervention resulted in increases in correct answers and certainty. Correct answers rose from 57% in the first questionnaire to 80% in the third ( p < 0.001). Certainty scores rose from 50% before the experiment to 70% one month afterwards ( p < 0.001). Despite 68% of learners having never taken part in an escape room game before, 79% liked this educational method. Conclusion This study built and tested a pedagogical escape room involving a high risk, professional, pharmacy process. The use of this pharmacy technology simulation had a positive impact on pharmacy staff theoretical knowledge.