Hydrazide-hydrazones as Small Molecule Tropomyosin Receptor Kina se A (TRKA) Inhibitors: Synthesis, Anticancer Activities, In silico ADME and Molecular Docking Studies

Abstract

Aims: The aim of the study was to search for new anticancer agents as TRKA inhibitors. Background: A series of new salicylic acid hydrazide hydrazones were synthesized and evaluated for their in vitro anticancer activities against lung (A549), ovarian (SK-OV-3), skin (SK-MEL-2), and colon (HCT15) cancer cell lines, and tropomyosin receptor kinase A (TRKA) inhibitory activities. Objective: In this study, we focused on the synthesis and anticancer properties evaluation of salicylic acid hydrazide hydrazones as TRKA inhibitors. The in vitro anticancer activities of hydrazone analogs were measured against four cancer cell lines, and the TRKA inhibitory properties were investigated using an enzyme assay to determine their modes of action. In silico molecular docking was conducted using the crystal structure of the TRKA receptor to study the interactions and modes of binding at its active site, and ligand-based target predictions were used to identify putative secondary enzymatic targets of the synthesized compounds. Additionally, pharmacokinetic properties, toxicity effects, and drug scores of the studied molecules were also assessed. Methods: A series of hydrazide hydrazones were prepared by means of a facile and straight-forward two-step reaction under soft reflux conditions from a methyl ester of substituted aromatic acids and hydrazine hydrate followed by the condensation with substituted aldehydes. In vitro cytotoxic properties of the synthesized compounds were screened against four human cancer cells using the SRB (sulforhodamine-B) colorimetric method. The TRKA inhibitory activity was measured by enzymatic assay. In silico ADME, drug score properties, docking studies, and ligand-based target prediction analyses were performed using Osiris Cheminformatics and AutoDock Vina, and SwissTargetPrediction bioinformatics software. Results: In vitro bioassays revealed that compound 6 exhibited the most potent broad-spectrum anticancer activities with IC50 values of 0.144, <0.001, 0.019, and 0.022 μM against A549, SK-OV-3, SK-MEL-2, and HCT15 cancer cells, respectively, followed by compounds 11, 3a, and 9. In TRKA inhibitory assays, compounds 3e and 11 demonstrated the highest potency with IC50 values of 111 and 614 nM, respectively. The results of docking studies on 3e and 11 with the active site of the TRKA receptor revealed that both compounds interacted as previously reported TRKA inhibitors with high docking scores. Conclusion: New salicylic acid hydrazide hydrazones were synthesized, and the most active compounds exhibited significant anticancer properties against A549, SK-OV-3, SK-MEL-2, and HCT15 cancer cells, suggesting to be good candidates for in vivo studies. The results obtained in the present study would help in the design and preparation of new hydrazidehydrazone analogs as potential TRKA inhibitors for cancer treatment.