Design, synthesis and biological evaluation of novel N-(3-amino-4-methoxyphenyl)acrylamide derivatives as selective EGFRL858R/T790M kinase inhibitors

Abstract

On the basis of N-(3-amino-4-methoxyphenyl)acrylamide scaffold, a series of novel compounds containing 3-substitutional-1-methyl-1H-indole, 2-substitutional pyrrole or thiophene moieties were synthesized and their in vitro antiproliferation activities against A549 and H1975 cell lines were evaluated. The results indicated that most of the compounds showed moderate to excellent antitumor activities. Especially, compounds 9a (A549 IC₅₀ = 1.96 μM, H1975 IC₅₀ = 0.095 μM), 17i (A549 IC₅₀ = 4.17 μM, H1975 IC₅₀ = 0.052 μM), 17j (A549 IC₅₀ = 1.67 μM, H1975 IC₅₀ = 0.061 μM) exhibited comparable antitumor activities and selectivity ratios compared to the positive control osimertinib (A549 IC₅₀ = 2.91 μM, H1975 IC₅₀ = 0.064 μM). In vitro inhibitory activities against EGFR kinases containing different mutations were also tested. Compound 17i showed remarkable inhibitory activity (with IC₅₀ value of 1.7 nM) to EGFR^L858R/T790M kinase and selectivity (22-folds compared to EGFR^WT kinase). Furthermore, acridine orange/ethidium bromide (AO/EB) staining assay, cell apoptosis assay, cell cycle distribution assay and wound-healing assay of the compounds 9a and 17i were performed on H1975 cell line. The results showed dose-dependent activities of the induction of apoptosis, G0/G1-phase arrestation and inhibition of migration, which were similar to the positive control osimertinib. Additionally, molecular docking analysis was performed to seek the possible binding mode between the selected compounds (9a, 17i-17j) and EGFR^L858R/T790M kinase. The results demonstrated that compound 17i is a promising candidate and worth further study.

Keywords: Antitumor activity; Docking study; EGFR-TKIs; Kinase selectivity; N-(3-amino-4-methoxyphenyl)acrylamide derivatives.