S-Klotho is perceived as a biomarker of healthy aging that has been shown to be inversely associated with cardiometabolic risk in elderly individuals. The aim of this study was to test if s-Klotho level is associated with cardiometabolic risk markers in younger healthy men in order to verify the possible role of s-Klotho level as an early marker of cardiometabolic risk. A cross-sectional study was conducted among 186 healthy men (Mage=35.33, SDage=3.47) from a Western urban population. Serum basal levels of s-Klotho, lipid profile, homocysteine, glycemia markers, C-reactive protein, liver transaminases and creatinine were evaluated. Also, blood pressure was measured and cardiometabolic risk score and homeostatic model assessment for insulin resistance (HOMA-IR) were calculated. Testosterone and cortisol levels, self-reported psychological stress, physical activity, smoking in the past, alcohol use and body adiposity were controlled for. We found no relationship between levels of s-Klotho and physiological markers of cardiometabolic risk in the studied population. The results were similar when controlled for adiposity, testosterone level, physical activity, alcohol use and smoking in the past. We suggest that s-Klotho level is not an early marker of cardiometabolic risk in younger middle-aged healthy men.