Office workers frequently display behavioral patterns marked by extended periods of sedentary and seated work. In this context, the implementation of a workplace exercise program has been shown to be effective in mitigating health-related risk factors. This study aimed to evaluate the impact of a 12-week workplace exercise program on the quality of life of employees whose primary occupational duties involve prolonged sitting. The final sample comprised 19 participants (6 females and 13 males), aged between 21 and 48 years (M = 32.26; SD = 9.03). Among these participants, 78.9% reported engaging in a hybrid work arrangement, 15.8% followed an on-site work regimen, and 5.3% worked remotely. Furthermore, 84.2% of participants reported sitting for 6 to 8 h per day, while 15.8% reported sitting for between 9 and 12 h daily. The quality of life of participants was assessed using the Medical Outcomes Study 36-Item Short Form Health Survey. A paired samples t-test was employed to examine differences between dependent variables. The results revealed a statistically significant improvement in general health (p < 0.01), vitality (p < 0.01), social functioning (p < 0.05), and the mental health summary measure (p < 0.04). Promoting physical activity through workplace exercise programs offers substantial benefits for workers’ health and quality of life, contributing to the prevention of occupational diseases.