Design and development of 2,7-disubstituted 6-methoxyquinazolin-4 (3H)-ones as potential anti-cancer agents possibly through downregulation of Wnt/β-catenin/TCF4 signaling pathway.

Abstract

Aim: A novel series of 2,7-disubstituted 6-methoxyquinazolin-4(3H)-one derivatives were designed, synthesized, and evaluated for in-vitro anti-cancer potential.

Methods: Compounds were docked, synthesiazed and then evaluated for in vitro anti-cancer activity using the sulforhodamine B assay in HCT116 and HepG2 cells. Apoptosis induction was analyzed via morphological changes, Hoechst 33342, and Annexin V/PI staining. The effect on β-catenin/TCF‑mediated transcriptional activity was assessed by TOPFlash/FOPFlash assay, TCF4 and β-catenin protein expression by immunocytofluorescence, and Wnt target genes (like c-MYC and Cyclin D1) mRNA levels by RT-PCR against HCT116 cells. Furthermore, In vitro anti-cancer potential was evaluated against primary human gallbladder cancer cells.

Results: The derivatives showed favorable binding with the active site residues on β-catenin and have the potential to disrupt the β-catenin/TCF4 interaction. Most of them have comparable anti-cancer activity to imatinib mesylate. Compound 42D, one of the potent compounds (IC₅₀: 3.89 μM in HCT116; 8.28 μM in HepG2), induced apoptosis, and significantly downregulated β-catenin/TCF4 signaling and downstream targets (c-Myc, Cyclin D1) in HCT116 cells and this could be one of the mechanisms by which it exerts its anti-cancer activity. It showed anti-cancer activity in primary gallbladder cancer cells (IC₅₀: 7.26 μM).

Conclusion: Compound (42D) represents a promising molecule as an anti-cancer agent against colon, hepatocellular, and gallbladder cancers targeting the Wnt/β-catenin/TCF4 signaling pathway.