We have reacted 2-dioxo-dihydropyrimidine-2,4(1H, 5H)-dione (thiobarbituric acid) with a different aromatic aldehyde in the presence of NaOH to give thiobarbituric acids analogs (1-29) which were characterized by EI-MS and 1H-NMR and subjected for α-glucosidase, thymidine phosphorylase and lipoxygenase inhibition studies. Analog 19 showed good α-glucosidase inhibitory potential with IC50 values of 37.61 ± 0.08 μM, while analogs 3, 6, 7, 9, 13, 15, 16, 18, 21-27 showed good to moderate α-glucosidase inhibitory potential when compared with standard acarbose having IC50 value 38.25±0.12 μM. Three analogs such as analog 20, 21 and 28 showed outstanding thymidine phosphorylase inhibitory potential with IC50 values 18.8, 40 and 6.9 μM respectively when compared with standard 7-deazaxanthine having IC50 value 41.0 ± 1.63 μM. In the case of lipoxygenase all the analogs were found inactive. Molecular docking studies were carried out to understand the binding interaction of the most active compounds.