In this work a series of hydrazone Schiff base compounds have been synthesized and characterized by FT-IR, 1H NMR, 13C NMR, and mass spectrometry. The structures of the target compounds designed to have 3,4,5-trimethoxy in acid hydrazide part and different substituents in the imine part. The antibacterial activity of the synthesized compounds has been studied against two gram positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis) and two gram negative bacteria (Escherichia coli and Klebsiella sp.), while the antifungal activity was studied against candida albicans fungi. The results revealed that most of the synthesized hydrazone derivatives exhibit a moderate antimicrobial activity when compared with the standard drug ampicillin. Molecular docking studies were carried out on the bacteria strain Staphylococcus aureus with target protein DHFR and its complex with trimethoprim (PDB ID: 2W9H).