Design, Synthesis and Biological Evaluation of Novel Substituted 4H-1,2,4-Triazole Derivatives as Anti-Cancer Agents

Abstract

Statistically, Colorectal carcinoma (CRC) has been found to be the third most deadly cancer with increasingly high rate of prevalence, incidences mortality, and morbidity as per the WHO reports. The tumor cells involve the aberrant Wnt-signalling pathway in more than 90% of CRC cases where tankyrase enzymes promote the proliferation of tumor through this Wnt signalling cascade. These undruggable targets were therefore selected for this work and extensive computational studies were carried out to design better tankyrase inhibitors. The use of 3D-QSAR studies, pharmacophore modeling and virtual screening, molecular docking and MD simulation studies using TNKS enzymes was employed to analyze, understand, and apply the knowledge of spatial atomic arrangements, key ligand-receptor interactions, their affinity, and their structural parameters to become druggable. Different softwares like Sybyl X1.2, Schrodinger’s glide module etc. were employed to carry out these extensive studies. The designed molecules were synthesized in reasonable quantities and structurally confirmed by their characterization using Mass spectroscopy, NMR spectroscopy (1H-NMR and 13C-NMR) and FTIR spectroscopy. Purity of each synthesized compound was assessed using HPLC analysis. All synthesized molecules were utilized to assess their potential as anticancer agents with the application of in vitro biological evaluation. The cytotoxicity and antiproliferative actions of each synthesized compounds at three different concentrations (i.e., 30 μM, 3 μM and 0.3 μM) were evaluated using Vero E6 cell lines, and other cancer cell lines including colorectal cancer cell lines (HT29 & SW480), lung cancer cell lines (A549) and breast cancer cell lines (MDA-MB231). Doxorubicin and XAV939 were used as reference standards to evaluate the comparative biological activity of these molecules. The best compounds found from this screening were subjected for apoptosis analysis using flow cytometry-based annexin-V-FITC/PI method at two different concentrations (i.e., 5 μM & 10 μM) to understand the nature of inhibition brought by the synthesized compounds against the internal standard XAV939. The overall conclusion from these extensive studies is that the currently designed compounds can be considered as the potential hits for the discovery of targeted therapy of CRC through inhibition of TNKS enzymes and Wnt-signalling cascade.