We have demonstrated that supaglutide has long-lasting effects on improving glucose tolerance of CD1 mice (Wang et al., 2010). In current study, we showed that supaglutide exerted dose-responsive effects in reducing glucose excursion in the euglycemic CD1 mice (Supplementary Figure 1). Furthermore, supaglutide at a dosage of 300 μg/kg was effective in improving glucose tolerance 216 h after a single drug-injection (Supplementary Figure 2). In order to determine the glucoregulatory effect of supaglutide on the diet-induced obese mice, we performed IPGTT and ITT in the obese mice, our studies showed that after the treatment for 4 weeks, the glucose tolerance and insulin sensitivity in the supaglutide-treated mice were significantly improved as compared to the control mice. Specifically, the treated obese mice showed lower level of fasting blood glucose and improved glucose excursion (Figure 9A) and improved insulin sensitivities (Figure 9B). When the glycemic response was expressed as AUC the changes were statistically different (Figure 9A', IPGTT: SPG vs. Ctrl = 1334.25 ± 19.86 vs. 1896.3 ± 53.58, p < 0.01; Figure 9B', ITT: SPG vs. Ctrl = 381.5 ± 7.63 vs. 648.13 ± 24.54, p < 0.01). Moreover, supaglutide was effective in reducing fasting blood glucose in the hyperglycemic db/db mice. Specifically, the drug-treated db/db mice exhibited a lower fasting blood glucose levels during 7 days' observations after a single dose injection (Supplementary Figure 3A). Interestingly, among doses tested, while supaglutide treatment showed dose-dependent effects on blood glucose lowering in the db/db mice, which was not observed in the context of its weight losing (i.e., lower dose tested showed better effects) (Supplementary Figure 3B). These results indicate that supaglutide exerted regulatory effects in energy homeostasis in the mice with established obesity and hyperglycemia.