Cancer is a terrible disease and second leading cause of death, behind cardio-vascular disease in the world. At present, there are three main methods of cancer treatment: surgery, radiation therapy and chemotherapy. Pyrimidineis a six-membered heterocyclic aromatic organic compound containing two nitrogen atoms at positions 1 and 3. Pyrimidine derivatives occupy a distinct and unique place in chemotherapy. The chemotherapeutic efficacy of pyrimidine derivatives is related to their ability to inhibit vital enzymes responsible for DNA biosynthesis as dihydrofolatereductase (DHFR), thymidylatesynthetase (TSase), thymidine phosphorylase (TPase) and reverse transcriptase (RTase). In the present study involves synthesis of 6-Substituted pyrimidine 2,4-diones derivatives. The synthesized compounds were subjected to antimicrobial activity against Gram negative E. coli (MTCC 40) and S. aureus (MTCC 87). The synthesized compounds possessed good to moderate antibacterial activity. Compounds 1, 2, 3a possessed good antibacterial activity when compared with standard however the compounds 2a, 2b, 2d, 1b, 1d, 1e possessed moderate activity. 1c, 1d, 1a, 2e, 2c were observed to be totally inactive compounds. The derivatives with electron withdrawing substituent on the phenyl ring at para position had poor activity in comparison to derivatives possessing no or electron donating substituents. The structures of the synthesized compounds were established by IR and NMR spectral studies.