High throughput screening of synthetic compounds against vital enzymes is the way forward for the determination of potent enzyme inhibitors.In-vitrohigh throughput library screening of 258 synthetic compounds (comp.1–258), was performed against α-glucosidase. The active compounds out of this library were investigated for their mode of inhibition and binding affinities towards α-glucosidase through kinetics as well as molecular docking studies. Out of all the compounds selected for this study, 63 compounds were found active within theIC50range of 3.2 μM to 50.0 μM. The most potent inhibitor of α-glucosidase out of this library was the derivative of an oxadiazole (comp.25). It showed theIC50value of 3.23 ± 0.8 μM. Other highly active compounds were the derivatives of ethyl-thio benzimidazolyl acetohydrazide withIC50values of 6.1 ± 0.5 μM (comp.228), 6.84 ± 1.3 μM (comp.212), 7.34 ± 0.3 μM (comp.230) and 8.93 ± 1.0 μM (comp.210). For comparison, the standard (acarbose) showedIC50= 378.2 ± 0.12 μM. Kinetic studies of oxadiazole (comp.25) and ethylthio benzimidazolyl acetohydrazide (comp.228) derivatives indicated that Vmaxand Km, both change with changing concentrations of inhibitors which suggests an un-competitive mode of inhibition. Molecular docking studies of these derivatives with the active site of α-glucosidase (PDB ID:1XSK), revealed that these compounds mostly interact with acidic or basic amino acid residues through conventional hydrogen bonds along with other hydrophobic interactions. The binding energy values of compounds25,228, and212were -5.6, -8.7 and -5.4 kcal.mol-1whereas RMSD values were 0.6, 2.0, and 1.7 Å, respectively. For comparison, the co-crystallized ligand showed a binding energy value of -6.6 kcal.mol-1along with an RMSD value of 1.1 Å. Our study predicted several series of compounds as active inhibitors of α-glucosidase including some highly potent inhibitors.