The present work reports the synthesis of 20 novel fluorine-containing quinoline and pyrimido[4,5 b]quinoline derivatives bearing a sulfonamide moiety. The new synthesized compounds were designed in compliance with the general pharmacophoric requirements for carbonic anhydrase (CA) inhibiting anticancer drugs, as this may play a role in their anticancer activity. All the newly synthesized compounds were evaluated for their in vitro anticancer activity against human breast cancer cell line (MCF7). Compounds 11 and 12 exhibited better activities than the reference drug doxorubicin (IC50 = 71.8 lM) with IC50 values of 52.6 lM and 67.3 lM, respectively. On the other hand, compounds 6, 10, and 13 showed IC50 values (71.8 lM, 69.8 lM, and 70.8 lM, respectively) comparable to that of the reference drug doxorubicin. In addition, docking of the synthesized compounds into human carbonic anhydrase isozyme II (hCA II) active site was performed in order to predict the affinity and the orientation of these compounds at the isozyme active site. Also, the most active compounds, 11 and 12, were selected and evaluated for their ability to enhance the cell killing effect of c-radiation.