Alkaline phosphatases (APs) are a class of homodimeric enzymes which physiologically possess the dephosphorylation ability. APs catalyzes the hydrolysis of monoesters into phosphoric acid which in turn catalyze a transphosphorylation reaction. Thiazoles are Nitrogen and sulfur containing aromatic heterocycles considered as effective APs inhibitors. Consequently, the current research presents the successful synthesis, spectroscopic characterization and in vitro alkaline phosphatase inhibitory potential of new thiazoles. Compound 5e was found to be the potent inhibitor of h-TNAP with IC50 value of 0.17 ± 0.01 µM. Additionally, compounds 5a and 5i were found to be highly selective toward h-TNAP with IC50 values of 0.25±0.01 µM and 0.21±0.02 µM respectively. Similarly in case of h-IAP, compound 5p served as the most potent inhibitor with IC50 value of 0.71 ± 0.02 µM. Structure activity relationship and molecular docking studies were performed to find out the binding modes of the screened compounds with the target site of tissue non-specific alkaline phosphatase (h-TNAP) and intestinal alkaline phosphatase (h-IAP). Accordingly, the most active inhibitors showed important interactions within the binding pockets of h-IAP and hTNAP, may be responsible for the inhibitory activity of the compound towards the enzymes. Therefore, the screened thiazole derivatives provided an outstanding platform for further development of alkaline phosphatase inhibitors.