This work is based on the synthesis of new ether derivatives bearing benzothiazole scaffold through multi-step reaction process. Initially, benzothiazole was prepared by refluxing 4-hydroxybenzaldehyde with amino thiophenol having sodium metabisulfite in DMF; subsequently the product was further refluxed with different substituted benzyl and alkyl bromide in acetone to get ether hybrids of benzothiazole in good yields. Structurally, these compounds were confirmed by means of 1H, 13C-NMR and mass spectrometry and evaluated for in vitro thymidine phosphorylase inhibitory activity. In the series, seven compounds attributed excellent inhibition against thymidine phosphorylase enzyme better than the standard. Similarly, three compounds showed good activity while two compounds were found inactive. Moreover, all these compounds showed no toxicity to normal human fibroblast cell line (BJ Cell Line). In addition, Gaussian calculations were performed on the 6-31++g(d,p) basis set to examine the 13 synthesized compounds at the B3LYP, HF, and M062X levels. Additionally, molecular docking calculations were performed on thymidine phosphorylase enzyme proteins (PDB IDs: 4EAD, 2WK6, and 4LHM), and ADME/T calculations were performed to investigate the effects and responses of these compounds in human metabolism.