Introduction: The inappropriate utilization of antibiotics and inadequate knowledge plays a major role in the development of resistance. Antibiotics resistance has become a public health concern worldwide. Objective: The main objective of the study was to assess the knowledge, attitude, and practices of pharmacy students regarding the use of antibiotics. Methodology: A cross-sectional survey-based study was conducted among the undergraduate students of pharmacy, from 1st year to 5th years in the Private University of Karachi. A 4-section Questionnaire was constructed based on the knowledge attitude and practices of students. 460 completely filled questionnaires were considered for the study and later evaluated with SPSS version 20.0. Results: The demographic distribution showed 27.55% male, and 72.45% female. While correctly answered responses found 93.9%. and 94.7% of students completely agreed improper use of antibiotics causes the resistance. 84.4% of students stated resistance is a worldwide serious problem. 79.4% of students stated inappropriate use can result in ineffective treatment, and 93.1% stated it increases the health cost. 80% practiced the use of antibiotics for a longer duration than prescribed. 31% poorly understood the disease condition, 32% of respondents agreed to use a previous prescription, 24.1% borrowed antibiotics from friends and family while 29.5% stated the easy availability of antibiotics on pharmacies. Conclusion: The study showed the inappropriate knowledge, easy availability of antibiotics as over the counter medicines, and their inappropriate practices regarding the use of antibiotics can result in the development of antibiotic resistance.