A series of 1-substituted-1H-tetrazole-5-thiol building blocks were synthesized (6a–h) and coupled by S-alkylation with 2-bromo-1-(4-(substituted phenylsulfonyl)piperazin-1-yl)ethanone (4a–c) using triethylamine in ethanol under reflux conditions. The structures of newly synthesized compounds were characterised by nuclear magnetic resonance and mass spectral data. Further, the hybrid compounds (7a–x) were screened for in vitro inhibitory effect on human cervical carcinoma (SiHA), breast adenocarcinoma (MDA-MB-235) and human pancreatic carcinoma (PANC-1) cell lines using sulforhodamine B assay. Antiproliferative assay revealed that most of the target compounds exhibited significant growth inhibitory activity (GI50≤0.1 µM) against all tested cancer cell lines compared to the reference drug. The most promising active compounds in this series were 7e and 7n, which displayed antiproliferative activity with GI50≤0.2 µM against the SiHa and MIDA-MB-231 cancer cell lines, whereas compounds 7g, 7l, 7p, 7s and 7t exhibited antiproliferative activity with GI50≤0.1 µM against the PANC-1 cell line. Thus the tetrazole-piperazinesulfonamide hybrid compounds could be potential leads for the development of new antiproliferative agents.