People practicing high-intensity interval exercise (HIIE) fasted during the morning hours under a lack of sleep. Such a habit may jeopardize the health benefits related to HIIE and adequate sleep. Fifteen habitually good sleeper males (age 31.1 ± 5.3 SD year) completed on a treadmill two isocaloric (500 kcal) HIIE sessions (3:2 min work:rest) averaged at 70% VO2reserve after 9-9.5 h of reference sleep exercise (RSE) and after 3-3.5 h of acute-partial sleep deprivation exercise (SSE). Diet and sleep patterns were controlled both 1 week prior and 2 days leading up to RSE and SSE. HIIE related performance and substrate utilization data were obtained from the continuous analysis of respiratory gases. Data were analyzed using repeated measures ANOVA with the baseline maximum oxygen uptake (VO2max) and body fat percentage (BF%) as covariates at p < 0.05. No difference was observed in VO2max, time to complete the HIIE, VE, RER, CHO%, and FAT% utilization during the experimental conditions. Whether attaining an adequate amount of sleep or not, the fasted HIIE performance and metabolism were not affected. We propose to practice the fasted HIIE under adequate sleep to receive the pleiotropic beneficial effects of sleep to the human body.