Background Despite the growing availability of smoking cessation apps, low engagement and cessation rates have remained a significant challenge. To address this issue, we used a user-centered design to iteratively develop a mobile app (MO) to provide comprehensive, tailored, and evidence-based content to support smokers in their quitting journey. Objective This study examined the acceptability, use, and preliminary efficacy of the MO app for smoking cessation. Specifically, we sought to understand smokers’ preferred features, engagement, and satisfaction with MO; identify concerns in using the app and ways to improve the app; and evaluate its smoking cessation outcomes. Methods Through 3 cohorts, we recruited 10, 12, and 85 adult smokers who attempted to quit smoking to pilot-test the MO app between December 2019 and July 2022. Participants were instructed to complete a baseline survey, interact with the app for 6 weeks, and fill in a postsurvey at week 6. Participants in cohort 3 completed an additional postsurvey at week 12. Participants’ app use was tracked and analyzed. The primary outcome measures were participants’ 7-day point prevalence abstinence at 6 and 12 weeks. Results Participants reported high levels of satisfaction with the MO app across all 3 cohorts, rating it between 4.40 and 4.76 on a scale of 5 for acceptability. Users engaged with app activities for an average of 89 to 159 times over 35 days. The most liked features of the app included “quit plan,” “tracking,” “reminders and notifications,” “MOtalks,” and “motivational quotes.” The 7-day point prevalence abstinence rate of the modified intention to treat population in cohort 3 was 58% at 6 weeks and 52% at 12 weeks. Those who interacted more frequently with app features and engaged with more diverse activities were more likely to maintain abstinence at weeks 6 and 12. For each additional time logged into the app, the odds of staying abstinent at week 12 increased by 5% (odds ratio [OR] 1.05, 95% CI 1.01-1.08). Participants who earned >5000 points during app use also had higher odds of quitting at both 6 weeks (OR 3.12, 95% CI 1.25-7.75) and 12 weeks (OR 4.65, 95% CI 1.83-11.76), compared with those who earned <5000 points. Conclusions Our study demonstrated that MO is a feasible mobile phone app with high acceptability and usability and can effectively deliver smoking cessation support to individuals who want to quit. Implications for developing and evaluating mobile phone apps for smoking cessation are discussed.