The study aims to determine the effect of enhancing knowledge and awareness of the WHO Access, Watch, and Reserve (AWaRe) antibiotics classification on hospital clinical staff’s knowledge, attitudes and antibiotic prescribing practices. A pre-post-intervention study design was employed. The intervention was an educational activity that involved teaching physicians and pharmacists about the AWaRe classification and the risk of antibiotic resistance. A questionnaire was administered to clinical staff pre-and post-intervention. In the pre-interventional stage, 78.5% of participants stated they had not heard about the AWaRe classification of antibiotics. After receiving the intervention: the knowledge regarding the meaning and purpose of AWaRe classification of antibiotics increased from 39.1% to 75.4%; the percentage of participants who agreed with following the AWaRe classification of antibiotics in their practice increased from 21.7% to 58.5%; and the percentage of participants who agreed that AWaRe classification of antibiotics can suggest safe choices of antibiotics increased from 56.5% to 90.8%. Hospital antibiotic use of the Access group increased by 6.6% from pre- to post-intervention. The use of the Watch group and Reserve group decreased post-intervention by 1.7%, and 43.1%, respectively. This study showed important gaps in knowledge and attitudes towards AWaRe, highlighting the need for increasing the awareness of the AWaRe tool amongst healthcare practitioners to ensure rational use of antibiotics.