Novel benzenesulfonamides as dual VEGFR2/FGFR1 inhibitors targeting breast cancer: Design, synthesis, anticancer activity and in silico studies

<p>In the current study, a new series of benzenesulfonamides <strong>6a-r</strong> was designed and synthesized as dual VEGFR-2 and FGFR1 kinase inhibitors with anti-cancer activity. The 4-trifluoromethyl benzenesulfonamide <strong>6l</strong> exhibited the highest dual VEGFR-2/FGFR1 inhibitory activity with IC₅₀ values of 0.025 and 0.026 µM, respectively. It showed a higher activity than sorafenib and staurosporine by 1.8- and 1.3-fold, respectively. Furthermore, compound <strong>6l</strong> was further tested on EGFR and PDGFR-β kinases showing IC₅₀ values of 0.106 and 0.077 µM, respectively. The target compounds were tested for their anticancer activity against NCI-60 panel of cancer cell lines at 10 µM concentration, where compound <strong>6l</strong> displayed the highest mean growth inhibition percent % (GI%) of 60.38 %. Compounds <strong>6a</strong>, <strong>6b</strong>, <strong>6e</strong>, <strong>6f</strong>, <strong>6 h-l</strong>, and <strong>6n-r</strong> revealed promising GI % on breast cancer cell lines (MCF-7, T-47D, and MDA-MB-231), and were subjected to IC₅₀ determination on these cell lines. The tested compounds showed a higher activity on T-47D and MCF-7 cell lines over MDA-MB-231 cell line compared to the used reference standard; sorafenib. Compounds <strong>6e</strong>, <strong>6 h-j</strong>, <strong>6l</strong> and <strong>6o</strong> revealed IC₅₀ values ≤ 20 µM against T-47D cell line, furthermore, they were found to be non-cytotoxic on Vero normal cell line. Furthermore, the effect of the most active compounds <strong>6i</strong>, and <strong>6l</strong> in T-47D cells on cell cycle analysis progression, cell apoptosis, and apoptosis markers was investigated. Both compounds arrested cell cycle progression at G1 phase, furthermore, they enhanced early and late apoptosis, as well as necrosis. The capability of compounds <strong>6i</strong>, and <strong>6l</strong> to induce apoptosis was further confirmed by their ability to raise BAX/BCl-2 ratio and caspase-3 level in the treated cells. Cell migration assay revealed that both compounds <strong>6i</strong> and <strong>6 l</strong> have anti-migratory effects compared to control T-47D cells after 24, and 48 h. Molecular docking studies for compounds <strong>6a-r</strong> on VEGFR-2 and FGFR1 binding sites showed that they exhibit an analogous binding mode in both target kinases which agrees with that of type II kinase inhibitors. </p>