Discovery of new symmetrical and asymmetrical nitrile-containing 1,4-dihydropyridine derivatives as dual kinases and P-glycoprotein inhibitors: synthesis, in vitro assays, and in silico studies

Abstract

Two new series of symmetric (1a-h) and asymmetric (2a-l) 1,4-DHP derivatives were designed, synthesised, and evaluated as anticancer agents. In vitro anticancer screening of target compounds via National cancer institute "NCI" revealed that analogues 1g, 2e, and 2l demonstrated antiproliferative action with mean growth inhibition percentage "GI%" = 41, 28, and 64, respectively. The reversal doxorubicin (DOX) effects of compounds 1g, 2e, and 2l were examined and illustrated better cytotoxic activity with IC₅₀ =1.12, 3.64, and 3.57 µM, respectively. The most active anticancer analogues, 1g, 2e, and 2l, were inspected for their putative mechanism of action by estimating their epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (HER-2), and Bruton's tyrosine kinase (BTK) inhibitory activities. Furthermore, the antimicrobial activity of target compounds was assessed against six different pathogens, followed by determining the minimum inhibitory concentration "MIC" values for the most active analogues. Molecular docking study was achieved to understand mode of interactions between selected inhibitors and different biological targets.

Keywords: 1,4-DHPs; P-glycoprotein; anticancer; antimicrobial; apoptosis; multidrug resistance (MDR); receptor tyrosine kinases (RTKs).

Figure 1.

Structure of some reported biologically active 1,4-DHP derivatives.

Figure 2.

The rationale for the molecular design of the target compounds as kinases and P-gp dual inhibitors.

Scheme 1.

Synthesis of the first series of symmetric achiral 1,4-DHP compounds, 1a-h. Reagents and conditions: (i) MeOH, heat under reflux, 24 h.

Scheme 2.

Synthesis of the second series, asymmetric chiral 1,4-DHP analogues 2a-l. Reagents and conditions: (i) MeOH, heat under reflux, 24 h.

Figure 3.

Summary of SAR for anticancer activity of target compounds besides structures of the most active analogues.

Figure 4.

Cytotoxicity (µM) of compounds 1 g, 2e, and 2 l on human HCT-116, HCT-116/ADR (without DOX), and HCT-116/ADR (with DOX) cells.

Figure 5.

% Potency of compounds 1 g, 2e, and 2 l against EGFR, HER-2, and BTK.

Figure 6.

Apoptosis assay: Impact of compound 1 g (upper left), compound 2e (upper right), compound 2 l (lower left), and DMSO (lower right) on the % of annexin V-FITC-positive staining in HCT-116 cells.

Figure 7.

Effect of compounds 1 g, 2e, and 2 l and DMSO on the percentage of HCT-116 cells stained positive for annexin V-FITC in apoptosis assay.

Figure 8.

Cell cycle analysis of HCT-116 cells treated with compound 1 g (upper left panel), compound 2e (upper right panel), compound 2 l (lower left panel), and DMSO (lower right panel).

Figure 9.

Outcome of compounds 1 g, 2e, and 2 l and DMSO on the percentage of HCT-116 cells cell cycle analysis.

Figure 10.

The 2D and 3D docking poses of compounds 1g, 2e, 2l and DOX interactions with P-gp (PDB ID: 3G60).

Figure 11.

The 2D and 3D docking poses of compounds 1g, 2e, 2l and lapatinib interactions with EGFR (PDB ID: 1M17).

Figure 12.

The 2D and 3D docking poses of compounds 1g, 2e, 2l and lapatinib interactions with HER-2 (PDB ID: 3RCD).

Figure 13.

The 2D and 3D docking poses of compounds 1g, 2e, 2l and lapatinib interactions with BTK (PDB ID: 4Z3V).