The present study investigated whether the caffeine supplementation for four days would induce tolerance to the ergogenic effects promoted by acute intake on physiological, metabolic, and performance parameters of cyclists. A double-blind placebo-controlled cross-over design was employed, involving four experimental trials; placebo (4-day)-placebo (acute)/PP, placebo (4-day)-caffeine (acute)/PC, caffeine (4-day)-caffeine (acute)/CC and caffeine (4-day)-placebo (acute)/CP. Fourteen male recreationally-trained cyclists ingested capsules containing either placebo or caffeine (6 mg∙kg−1) for 4 days. On day 5 (acute), capsules containing placebo or caffeine (6 mg∙kg−1) were ingested 60 min before completing a 16 km time-trial (TT). CC and PC showed improvements in time (3.54%, ES = 0.72; 2.53%, ES = 0.51) and in output power (2.85%, ES = 0.25; 2.53%, ES = 0.20) (p < 0.05) compared to CP and PP conditions, respectively. These effects were accompanied by increased heart rate (2.63%, ES = 0.47; 1.99%, ES = 0.34), minute volume (13.11%, ES = 0.61; 16.32%, ES = 0.75), expired O2 fraction (3.29%, ES = 0.96; 2.87, ES = 0.72), lactate blood concentration (immediately after, 29.51% ES = 0.78; 28.21% ES = 0.73 recovery (10 min), 36.01% ES = 0.84; 31.22% ES = 0.81), and reduction in expired CO2 fraction (7.64%, ES = 0.64; 7.75%, ES = 0.56). In conclusion, these results indicate that caffeine, when ingested by cyclists in a dose of 6 mg∙kg−1 for 4 days, does not induce tolerance to the ergogenic effects promoted by acute intake on physiological, metabolic, and performance parameters.