BackgroundAntibiotics resistance is a serious problem around the world, which has been increasing in the last century due to misuse of antibiotics. Recently, Saudi Arabia enforced the Executive Regulations of Health Practice Law that prohibits dispensing antibiotics without prescription.Aim(1) To assess the association between the knowledge and attitude toward the recent enforcement of the antibiotic restriction Law and the antibiotic use among Saudi population.(2) To assess the pattern of antibiotic use before and after the enforcement of the Law.MethodAn online cross-sectional study was conducted among residents of Saudi Arabia.ResultsThe study was carried out among 570 Saudi participants; the result shows no considerable difference before and after law enforcement in purchasing and use of antibiotics. During the six months before the law implementation, antibiotics were obtained by 42 (7%) through purchase from the pharmacy without a prescription, 23 (4%) through remains of previously used containers, and 7 (1%) through a relative or a friend; while during the six months after the law implementation it was 31 (5%), 22 (4%) and 7 (1%), respectively. Only 48% agreed that they always received advice from a doctor, nurse or pharmacist on how to use your antibiotics. 32% have no knowledge about antibiotic resistance or drug resistance or antibiotic-resistant bacteria. More than two third (75%) were aware of the recent enforcement of the law that prohibits dispensing antibiotics without prescription. Among the participants, 91 (16%), 285 (50%) and 194 (34%) were expressed negative, neutral and positive attitude towards the law, respectively.ConclusionIn spite of the recent enforcement of the Law, there is still misuse of antibiotics, which indicate need for educational programs and campaigns to improve the knowledge of the population.