Synthesis, cytotoxic and anti-microbial screening of novel thiobarbituric acid incorporated pyrazole derivatives were performed. Vilsmeier-Haack reaction of different phenyl hydrazones 1(a-e) afforded pyrazole -4- carbaldehydes 2(a-e) in good yields. Knoevenagel condensation of compounds 2(a-e) with thiobarbituric acid gave a series of 5-ylidene derivatives 3(a-e) in reasonable yields. The synthesized compounds were characterized with the help of IR,1HNMR and mass spectral data. The compounds were tested for cytotoxic activity against Vero, MCF-7 and HCT-116 cell lines. Among the tested compounds, 5- ((3 -(4-hydroxyphenyl)-1-phenyl-1H-pyrazol-4- yl)methylene)-2-thioxodihydropyrimidine-4,6-(1H,5H)-dione (3d) was found to be most active molecule with the activity against both MCF-7 and HCT-116 cell lines with IC50 values of 14.0 μM and 18.12 μM. In anti-microbial screening, none of the compounds exhibited anti-bacterial activity.