In an effort to develop new antimicrobial agents, a novel series of 2-{[4-(substituted phenyl)amino]methyl)-1H-isoindole-1,3(2H)-dione derivatives 2-17a,b was synthesized starting from 2-({[4-(bromoacetyl)phenyl]amino}methyl)-1H-isoindole-1,3 (2H)-dione 1 by introducing different hetero-cyclic moieties, such as thiazole, thiazolidinone, azetidinone, furan and pyrazole. The structures of all the synthesized compounds have been elucidated by means of IR, 1H NMR, mass spectroscopic data and elemental analysis. Most of the synthesized compounds were screened for their antimicrobial activity by means of agar well diffusion assay while the minimal inhibitory concentrations of the active compounds were then assessed utilizing broth dilution method. Derivative 2-[(4-{[(1,3-Dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl] amino} phenyl)-2-oxoethyl]propane dinitrile 15 was the most potent compound.