S-Alkylated quinazolin-4(3 H)-ones as dual EGFR/VEGFR-2 kinases inhibitors: design, synthesis, anticancer evaluation and docking study

Abstract

Dual targeting by a single molecule has emerged as a promising strategy for fighting cancer. In this study, a new set of 2-thioquinazolin-4(3H)-ones as potential anti-cancer surrogates endowed with dual EGFR/VEGFR-2 kinases inhibitory activities were synthesized. The anti-tumor potency of the newly synthesized candidates 4-27 was evaluated against a panel of four cancer cell lines. The prepared candidates 4-27 showed comparable activity to that of the standard drug sorafenib. For instance, compound 4 (IC₅₀ = 1.50-5.86 μM) and compound 20 (IC₅₀ = 4.42-6.39 μM) displayed superior potencies against all cell lines compared to sorafenib (IC₅₀ = 5.47-7.26 μM). Dual EGFR/VEGFR-2 inhibitory activities of the most active analogues (4, 11, and 20) were investigated. Compound 4 showed comparable EGFR/VEGFR-2 inhibitory activity to the used control drugs. Flow cytometric analysis indicates that the most potent analogue 4 stopped the cell cycle at the G1 phase and induced 46.53% total apoptosis in HCT-116 cells that was much more powerful than the untreated cells with 2.15% apoptosis. Molecular docking and dynamic simulations of 4, 11, and 20 with EGFR and VEGFR-2 were performed to examine the binding mode and interaction within the enzyme binding pockets.

Fig. 1. The design strategy of our target compounds is guided by previously reported antitumour drugs/agents.

Scheme 1. Synthesis of quinazoline analogues 4–27 with various S-linked fragments.

Fig. 2. Summary of structure–activity correlation of target s-alkylated quinazoline-4(3H)-ones 4–27.

Fig. 3. Effect of 4 (left panel) and DMSO as a control (right panel) on DNA-ploidy flow cytometric analysis of HCT-116 cells after treatment for 24 h.

Fig. 4. Compound 4 affects the percentage of annexin V-FITC-positive staining in HCT-116 cells. The cells were treated with DMSO as control and 4 for 24 hours. Dead (necrotic) cells exist in the Q1 quadrant; late apoptosis is represented in the Q2 quadrant; live cells are shown in the Q3 quadrant; early apoptosis is depicted in the Q4 quadrant.

Fig. 5. Total, late and early apoptosis induced by 4 compared to DMSO control.

Fig. 6. Visual representation of molecular interactions within the EGFR active site. This 3D model showcases the comparative binding patterns of investigated 4 (in pink), 11 (in yellow), 20 (in cyan), and reference compound LPB (in metallic hue). Each compound exhibits distinct conformational orientations and interactions within the kinase domain, highlighting their potential differential impacts on EGFR's (PDB: 1xkk) enzymatic activity and signaling pathways.

Fig. 7. Visual representation of molecular interactions within the VEGFR-2 active site. The 3D model showcases the binding patterns of the investigated 4 (in pink), 11 (in yellow), 20 (in cyan), and the reference compound SRB (in green). Each compound exhibits unique conformational orientations and interactions within the kinase domain, highlighting their potential differential impacts on VEGFR-2's enzymatic activity and signaling pathways (PDB: 3wze). This visual representation offers valuable insights into the compounds' molecular mechanisms and potential effects on VEGFR-2 and its associated signaling pathways.

Fig. 8. Root Mean Square Deviation (RMSD) trends for EGFR and VEGFR2 in its apo form (depicted in red and black, respectively) and when bound to 4 with EGFR and VEGFR2 (illustrated in blue and green, respectively) over the simulation time. The plot highlights the dynamic conformational changes of EGFR and VEGFR2 in both states.