During some trail running races, athletes have to carry hydration support, food, and technical and safety equipment, which generates an additional load that must be mobilized during the race. The aim of the present study was to determine the physiological responses to overload running and the effect they may have on metabolic zones. Seventeen well-trained male trail runners (n = 17) completed three maximal treadmill tests with weighted vests at 0%, 5%, and 10% of their body mass (L0, L5, and L10). Their gas exchange was monitored to assess their ventilatory thresholds 1 (VT1) and 2 (VT2), maximal fat oxidation zone (FatMax), and peak oxygen consumption (VO2peak). Their heart rate (HR), power, and velocity (V) were tracked to compare their behavior. One-way ANOVA showed significant differences in the V (p < 0.001; ηp2 = 0.4620) as a limitation for reaching the peak velocity (Vpeak), with a significant decrease in the Vpeak with the L10 compared to the L0 (p = 0.002) and L5 (p = 0.004). In addition, one-way ANOVA showed significant differences in the peak absolute power (p < 0.001; ηp2 = 0.468) among the groups, detecting higher power production between the L10 and L0 (p < 0.001) and between the L10 and L5 (p = 0.015). Loads higher than L5 could generated important physiological and mechanical modifications, while a load of L5 managed to maintain the working conditions without overloading. These insights shed light on nuanced strategies for optimizing performance and endurance, offering valuable considerations for athletes seeking to enhance their training regimens during overload conditions.