New S- and N-alkyl functionalized bis-1,2,4-Triazolyl-based derivatives as potential dual EGFRWT and EGFRT790M inhibitors: Synthesis, anti-proliferative evaluation, molecular docking study and ADMET studies

Abstract

A new series of 2-(s-triazolylsulfanyl) moiety connected with N-arylacetamides 8a-e, 1-aryl ethanones 10a-c, acetate 12a, acetonitrile 12b, and 2-propane 12c were synthesized via the reaction of 5,5′-(butane-1,4-diyl)bis(4-phenyl-4H-1,2,4-triazole-3-thiol) (6) with 2‑chloro-N-arylacetamides 7a-e, phenacyl bromides 9a-c, ethyl chloroacetate 11a, chloroacetonitrile 11b, or 2-bromopropane 11c under PTC conditions. Mannich bases 14a,b were synthesized by reacting compound 6 with formaldehyde and morpholine 13a or piperidine 13b. The structure of the new products has been characterized by IR, NMR, and their elemental analyses.

These newly designed scaffolds were designed as efficient anti-proliferative EGFR^WT and EGFR^T790M kinase inhibitory agents. Scaffolds 8c, 10b, and 12c were established as the most potent anti-proliferative derivatives on various tested cancer cell panels with highly remarkable IC₅₀ values compared to the reference drugs erlotinib and doxorubicin. Moreover, compounds 8c, 10b, and 12c demonstrated a favorable safety profile when tested against the normal WI-38 line, besides their exceptional selectivity as a dual kinase inhibitor, particularly targeting EGFR^WT and EGFR^T790M, where they were more potent than erlotinib and gefitinib, with IC₅₀ values of (0.30 and 11.47 µM), (0.38 and 11.57 µM), (0.33 and 13.12 µM), and (0.37 and 21.56 µM), respectively. Furthermore, its apoptotic triggering capability was investigated by evaluating apoptotic markers: Caspases-3, Caspases-9, Bcl-2, Bax, and Bax/Bcl-2 ratio expression levels.

The docking outcomes of the designed scaffolds 8c, 10a, 12c, and 14b in the ATP-binding sites of both EGFR^WT and EGFR^T790M agreed with the in vitro results. Besides, Molecular dynamics simulations via iMODs server evaluated the stability of protein-8c complexes. Also, the in silico ADME properties’ examination of the most promising EGFR inhibitory compounds 8c, 10b, and 12c via the egg-boiled method clarified acceptable lipophilicity, GIT absorption, and blood-brain barrier penetration. So, our designed analogs, specifically 8c and 12c, own prospective anti-proliferative and dual EGFR^WT and EGFR^T790M kinases’ inhibitory properties, making them efficient candidates for further therapeutic amelioration in the future.