In our current study thirteen new 2,4-dinitrophenyl hydrazone derivatives 1–13 have been evaluated for alpha amylase activity. The molecular docking results indicate that compounds potentially bind in the catalytic site of the enzyme with excellent result. Molecular Operating Environment (MOE) software was used for docking study. 2,4-Dinitrophenyl hydrazone 1-13 have been obtained under reflux conditions by reacting dinitrophenyl hydrazine in methanol with different aromatic as well as aliphatic aldehydes in the presence of acetic acid act as a catalyst. The current results have shown that compounds 5 (IC50 =12.16µg/mL), 6 (IC50 =15.03µg/mL), and 12 (IC50 =16.42 µg/mL) have been found to be the more potent alpha amylase inhibitors as compared to the standard acarbose (IC50 = 42.47µg/mL). These compounds may provide better leads for alpha amylase inhibitor and further assessment of these compounds can be of great help in the discovery of new antidiabetic drugs.