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Abstract Background Teleworking is associated with lifestyle risk factors, such as insufficient physical activity (PA) and increased sedentary time (ST); however, effective interventions tailored to teleworkers are lacking. We aimed to evaluate the effectiveness of a 12-week multicomponent occupational lifestyle intervention on daily step counts among Japanese teleworkers. Methods This 12-week, two-arm, parallel-group, cluster randomised controlled trial conducted across 12 clusters in six Japanese companies involved 310 teleworkers (mean age 43.0 years; 72.6% men) randomized to the intervention (6 clusters, n=156) or a waitlist control group (6 clusters, n=154). The multicomponent occupational lifestyle intervention included individual (online lectures, feedback, and email messages), physical (posters and a pop-up), and organizational (encouraging messages from an executive) strategies. The primary outcome was the change in daily step counts, measured using an accelerometer over 14 consecutive days at baseline and at week 12. Analyses were based on the intention-to-treat approach using a generalised estimating equation model. Findings The intervention group showed an adjusted mean change in daily step counts of +219 steps (95% confidence interval [CI] −165 to 604), compared with +188 steps (95% CI −183 to 560) in the control group. The adjusted between-group difference was +55 steps (95% CI −550 to 660; p=0.844). No significant effects on the secondary outcomes (ST, light PA, or moderate-to-vigorous PA) were observed. Interpretation The multicomponent occupational lifestyle intervention did not significantly increase daily step counts among Japanese teleworkers. Therefore, further studies should be done on tailored interventions for teleworkers. Research in context Evidence before this study Teleworking has increased globally, particularly following the coronavirus disease pandemic, and has been associated with reduced physical activity and increased sedentary behaviour, both of which are risk factors for cardiovascular disease. Previous studies have also reported that telework environments may contribute to musculoskeletal and other somatic symptoms. Multicomponent interventions in traditional office settings can effectively increase physical activity and reduce sedentary time. These interventions commonly employ strategies at multiple levels of the social–ecological model, including individual approaches (e.g., lectures or incentives), interpersonal approaches (e.g., team-based activities), environmental modifications (e.g., office rearrangements or sit-stand desks), and organisational support (e.g., leadership encouragement). The applicability of such interventions to teleworking populations remains unclear because teleworkers face distinct challenges such as social isolation, blurred work–life boundaries, and heterogeneous home working environments. These contextual differences highlight the need for interventions specifically tailored to teleworkers. Added value of this study We evaluated the effectiveness of a multicomponent occupational lifestyle intervention specifically designed for teleworkers, a population whose work environment differs substantially from traditional office settings. Our findings provide novel evidence that can inform the development of more targeted strategies to promote physical activity in evolving work environments. This study also provides objective measurements of physical activity using accelerometers, enabling detailed evaluation of step counts, sedentary time, and different activity intensities among teleworkers. Additionally, we used daily diaries to distinguish activity patterns across workdays, weekends, teleworking days, and commuting days, providing a nuanced understanding of behavioural patterns in remote work contexts. Implications of all the available evidence Our findings indicate that low-cost, remotely delivered multicomponent interventions may be insufficient to produce meaningful behavioural change among teleworkers. Similar strategies, including individual, physical, and organisational components, have been effective in traditional workplace interventions; however, their implementation in teleworking environments may not adequately address the specific challenges faced by remote workers. In particular, the lack of strong sociocultural support mechanisms, such as team-based step competitions or workplace champions, may limit engagement and reduce the effectiveness of such programmes. Therefore, further investigation is needed to explore more diverse and targeted intervention strategies, identify the specific needs and behavioural patterns of teleworkers, and apply more precise eligibility criteria to better address heterogeneity within teleworking populations.