Sorghum is used to provide good quality gluten-free products due to phytochemicals and low glycemic index (GI). This study aimed to determine the chemical composition, the antioxidant activity and capacity, and the glycemic and insulinemic responses of gluten-free (GF) sorghum bread. GF bread samples were produced with three different sorghum genotypes. The samples were analyzed for chemical composition, resistant starch and dietary fiber content; antioxidant activity by ORAC; antioxidant capacity by FRAP; GI; and insulinemic responses. This double-blind, crossover, randomized clinical trial was conducted with 10 healthy men aged 28.0 ± 4.9 years (77.6 ± 11.7 kg and 24.2 ± 2.3 kg/m2). All sorghum bread showed significantly more fiber than rice bread (control). Brown sorghum bread was classified as low GI, bronze and white as medium GI, and control as high GI. Brown sorghum bread presented a low carbohydrate content, a significant amount of fiber, and a significantly lower 3 h AUC glucose response than those of the control, aside from the highest antioxidant activity value (p ≤ 0.001). Therefore, brown sorghum was superior to other genotypes analyzed in this study, and its production should be encouraged to provide gluten-free products with a better nutritional profile. More research is required to explore the effects of different sorghum genotypes in food products on human health.