This pilot study investigated the impact of breaking up prolonged sitting with light-intensity walking on postprandial glucose responses and sleep architecture. In a randomized, counterbalanced, crossover design, six healthy males completed a sitting condition and an active condition (sitting interrupted with light-intensity walking) for three consecutive days, following 5-h sleep opportunities at night. Postprandial glucose response and sleep (time spent in all stages) was assessed. Breaking up prolonged sitting with light-intensity walking did not affect postprandial glucose responses in sleep-restricted participants; however a small increase (~9 min) in slow-wave sleep was observed.