The aim of this study was to examine acute physiological responses, physical fitness parameters and time-motion characteristics associated with a 4-a-side small-sided game in amateur level players. Sixteen male football players (age 23.9±4.2 years) completed one 4-a-side small-sided game with the aim of maintaining ball possession as long as possible. The participants were monitored for external load and physiological parameters, and tested before and after for physical fitness assessment. A Student’s paired t-test was conducted to determine the differences in physical fitness assessment. A repeated measures analysis of variance (ANOVA), with Bonferroni post-hoc test, was used to determine the dynamics of physiological parameters in players, their RPE and time-motion characteristics of SSG. A significant difference was found in 20 m sprint time between before and after the SSG (Δ=+1.3%). No significant differences were found between bouts or recovery periods for THb and SmO2, nor between HR-related variables (ES=.005-.383). Compared to RPE in bout 1, greater values were observed in bouts 2, 3, and 4 (Δ=+5.3%, p=.008, ES=.40; Δ=+9.6%, p=.002, ES=.98; Δ=+15.1%, p=.000, ES=1.29; respectively). No significant differences were found between bouts for time-motion characteristics. The results demonstrated that RPE responses increased throughout bout periods, whereas the rest of physiological parameters were maintained over the entire protocol. Due to possible fatigue accumulation, physical fitness performance was impaired (sprint) and the external load was reduced over the course of the protocol.