Seven new thiophene hydrazide derivatives 3a-e, 5, and 7 were prepared through the coupling of active methylene of compound 1 with diazonium salts of aromatic amines 2a-e, and heterocyclic amines 4 and 6 at 0-5 ^oC in pyridine. The studied compounds 1, 3a-e, and 5 could exist in two possible tautomeric forms, which are the enol and keto tautomer. The optimized molecular structures and calculation of the total energies of both tautomers revealed that the enol tautomer is energetically lower than its corresponding keto form. A prediction study for the biological activities of synthesized thiophene hydrazide derivatives 3a-e and 5 was performed via using PASS online software, which displayed promising activities in the treatment of Posttraumatic stress disorder, as Phosphodiesterase X inhibitor (3a, 3b), and as Sarcosine oxidase inhibitor (3d). In addition, DFT calculations showed that compounds 3a, 3b, and 3d have chemical activity among all the newly synthesized compounds due to their lower band gaps.