Design, synthesis, biological evaluation, in silico ADME prediction and molecular docking of pyrazole-benzamides as multitargeting protien kinase inhibitors

Abstract

In the present study, N-(1-(2-bromobenzoyl)-4-cyano-1H-pyrazol-5-yl) halogenated benzamides (5a-h) were prepared and evaluated for their in vitro anticancer activity against three cancer cell lines, namely hepatic cancer (HepG-2), breast cancer (MCF-7) and colon cancer (HCT-116). The derivatives 5a-d and 5 h revealed the most promising growth inhibition effect, with IC50 values ranging from 0.005133 ± 0.000042 to 0.022249 ± 0.000069 μM, compared to doxorubicin, which had IC50 values of 0.004635 ± 0.000002, 0.004587 ± 0.000029, and 0.004064 ± 0.000002 µM respectively. The potential ADME properties of 5a-h were investigated using calculations. Compounds 5a-h have good oral bioavailability and gastrointestinal absorption but cannot penetrate the blood-brain barrier. According to ADME results, 5b has a tumorigenic and reproductive effect. Furthermore, the derivatives 5a, 5b, and 5d produced promising safety profiles on the WI-38 normal cell line with IC50 values of 0.045374 ± 0.002593, 0.115714 ± 0.006732, and 0.058241 ± 0.003411 μM, respectively, compared to doxorubicin, which had IC50 value of 0.038234 ± 0.00223 µM. Moreover, utilizing afatinib and sorafenib as positive controls, compounds 5a, 5b, and 5d were further assessed as in vitro multitargeting candidates towards EGFRWT, VEGFR-2, and B-RAFWT. For the purpose of figuring out potential ligand-protien interactions and stability, in silico molecular docking simulations against VEGFR-2, EGFRWT, and B-RAFWT were run. The results exhibited that compound 5b was the most potent compound, producing an inhibition effect on EGFRWT and B-RAF WT kinases that was approximately equipotent to that obtained by the reference drugs afatinib and sorafenib with IC50 values of 0.000140 ± 0.000009 and 0.000158 ± 0.000009 µM, respectively (IC50 Afatinib: 0.000086 ± 0.000004, IC50 Sorafenib: 0.000097 ± 0.000006) The molecular docking study confirmed the biological result. Moreover, compound 5b arrested the HCT-116 cell cycle at the G1/S phase and showed a good early and late apoptotic induction effect within HCT-116 cells.