The discovery of novel alpha-glucosidase inhibitors and anti-diabetic candidates from natural or natural-derived products represents an attractive therapeutic option. Here, a collection of acetylphenol analogues derived from paeonol and acetophenone were synthesized and evaluated for their alpha-glucosidase inhibitory activity. Most of derivatives, such as 9a-9e, 9i, 9m-9n and 11d-1e, (IC50 = 0.57 +/- 0.01 mu M to 8.45 +/- 0.57 mu M), exhibited higher inhibitory activity than the parent natural products and were by far more potent than the antidiabetic drug acarbose (IC50 = 57.01 +/- 0.03 mu M). Among these, 9e and 11d showed the most potent activity in a noncompetitive manner. The binding processes between the two most potent compounds and alpha-glucosidase were spontaneous. Hydrophobic interactions were the main forces for the formation and stabilization of the enzyme acetylphenol scaffold inhibitor complex, and induced the topography image changes and aggregation of alpha glucosidase. In addition, everted intestinal sleeves in vitro and the maltose loading test in vivo further demonstrated the alpha-glucosidase inhibition of the two compounds, and our findings proved that they have significant postprandial hypoglycemic effects.