High-intensity interval training (HIIT) has become a popular exercise approach due to its potential to elicit significant physiological adaptations. However, limited research has compared the effects of different HIIT modalities on fitness parameters in middle-aged males. This study aimed to determine the effect of different HIIT on V̇O<inf>2max</inf>, strength, flexibility, and body fat percentage among middle-aged males. Thirty sedentary males aged 40-50 years were randomly assigned to HIIT-Weight Training Circuit (WTC), HIIT-Shuttle Run (SR), or HIIT-Bodyweight Exercise (BWE) groups. Training consisted of 20 sessions over 5 weeks of 4×4 minutes at 90-95% HR<inf>max</inf> with 3 minutes recovery at 70% HR<inf>max</inf> between intervals. Outcome measures including V̇O<inf>2max</inf>, strength, flexibility, and body fat percentage were assessed pre- and postintervention. The result for the Two-Way Mixed ANOVA indicated there was no significant interaction effect between the three training groups. There was no significant main effect for the three training groups, whereas there was a significant effect for the repeated measure time existed. A significant improvement in V̇O<inf>2max</inf>, strength, flexibility has been shown between post-test and pre-test for the HIIT-SR, HIIT-WTC, and HIIT-BWE group. Whereas A significant decrease has been shown between post and pretest for all the groups in body fat percentage. This research findings suggested that different forms of HIIT can elicit differential adaptation in middle-aged males. Incorporating different form of HIIT as HITT-SR, HITT-WTC, and HITT-BWE into exercise program may be an effective way to systematically improve V̇O<inf>2max</inf>, strength, flexibility, and body composition in middle-age male population. These findings provide guidance on HIIT prescription according to fitness goals.