Cancer is the disease with the highest mortality. Drug studies contribute to promising treatments; however there is an urgent need for selective drug candidates. Pancreatic cancer is difficult to treat and the cancer progresses rapidly. Unfortunately, current treatments are ineffective. In this study, ten new diarylthiophene-2-carbohydrazide derivatives were synthesized and evaluated for their pharmacological activity. The 2D and 3D anticancer activity studies suggested the compounds7a,7d, and7fwere promising. Among these,7f(4.86 µM) showed the best 2D inhibitory activity against PaCa-2 cells. Compounds7a,7dand7fwere also tested for their cytotoxic effects on healthy cell line but only compound7dshowed selectivity. Compounds7a,7d, and7fshowed the best 3D cell line inhibitory effect according to spheroid diameters. The compounds were screened for their COX-2 and 5-LOX inhibitory activity. For COX-2, the best IC50value was observed for7c(10.13 µM) and all compounds showed significantly lower inhibition compared to standard. In the 5-LOX inhibition study, compounds7a(3.78 µM),7c(2.60 µM),7e(3.3 µM), and7f(2.94 µM) demonstrated influential activity compared to standard. Regarding molecular docking studies, binding mode of compounds7c,7e, and7fto the 5-LOX enzyme were non-redox or redox types, but not the iron-binding type. As dual inhibitors of 5-LOX and pancreatic cancer cell line,7aand7fwere identified as the most promising compounds.