A series of hesperidin derivatives were prepared and identified by IR, 1H NMR, and MS spectra. All the synthetic compounds had been evaluated in vitro and in vivo based on α-glucosidase inhibition, glucose consumption of HepG2 cells and blood glucose level in streptozotocin-induced diabetic mice. The results revealed that all the compounds exhibited the anti-hyperglycemic activities. Compounds 3 and 7a had the excellent inhibition on α-glucosidase as compared to acarbose. Compound 3 could improve the glucose consumption of HepG2 cell and apparently reduce the blood glucose level of the streptozotocin-induced diabetic mice. Compound 3 exhibited the promising anti-hyperglycemic activity. A series of hesperidin derivatives were prepared and identified by IR, 1H NMR, and MS spectra. These compounds were evaluated in vitro and in vivo based on α-glucosidase inhibition, glucose consumption of HepG2 cells, and blood glucose level in streptozotocin-induced diabetic mice. The results revealed that all the compounds exhibited anti-hyperglycemic activities. The inhibition at 10−3M of compounds 3 and 7a on α-glucosidase were 55.02% and 53.34%, respectively, as compared to 54.80% by acarbose. Treated by compound 3 and the reference drug metformin, glucose consumption of HepG2 cell were 1.78 and 2.11mM, respectively. After the streptozotocin-induced diabetic mice were oral administrated with compound 3 at 100mgkg−1d−1 for 10days, the blood glucose level of 3 treated mice (13.23mM, P<0.05) showed significant difference when compared to model control (23.03mM). Thus, compound 3 exhibited promising anti-hyperglycemic activity.