Design, synthesis, and <i>in silico</i> studies of new benzofuran–pyrazole hybrids as multi-kinase inhibitors with potential antiproliferative activity

A new series of benzofuran-pyrazole-based analogues, conjugated with different substituted aromatic and heterocyclic ring systems featuring the pharmacophoric fragments of protein kinase suppressors, 3a-d and 4a-d was synthesized as potential antiproliferative agents. All the new analogues were selected by the NCI to screen their antiproliferative activity against sixty human cancer cell lines (NCI60). The 1<i>H</i>-benzo[<i>d</i>]imidazole derivative 3d demonstrated the highest percentage inhibition for various cancer cell lines and advanced to the five-dose assay. It showed potent anti-proliferative activity against various types of cancer lines with GI₅₀ values ranging from 0.33 to 4.87 μM and LC₅₀ values exceeding 100 μM against the majority of the tested cell lines, confirming its non-lethal effects. Additionally, 3d exhibited multi-targeting PK-suppression activity against B-Raf (V600E), c-Met, Pim-1, EGFR (WT), and VEGFR-2, with IC₅₀ values of 0.078 ± 0.004, 0.405 ± 0.017, 1.053 ± 0.046, 0.177 ± 0.007 and 0.275 ± 0.011 μg mL^-1, respectively. Moreover, 3d caused cell cycle arrest at the G0-G1 phase besides early and late apoptosis in MCF-7 cancer cells. <i>In silico</i> molecular docking and ADMET studies were performed on 3d to determine its expected binding interactions with the key regions in the kinase domains, as well as to ascertain its risks of human toxicity, drug-likeness traits, and oral bioavailability.