Insilico and in vitro Approach for Design, Synthesis, and Anti-proliferative Activity of Novel Derivatives of 5-(4-Aminophenyl)-4-Substituted Phenyl-2, 4-Dihydro-3H-1, 2, 4-Triazole-3-Thione
*Corresponding Author E-mail: Kubbaammar1963@gmail.com
A series of three 5-(4-aminophenyl)-4-substituted phenyl-2,4 dihydro-3H-1,2,4-triazole-3-thione analogs were designed, synthesized and investigated for anti-proliferative activity using in-silico and in vitro studies. The synthesis started by reaction of ethyl-4-aminobenzoate with hydrazine hydrate in absolute ethanol, to yield intermediate 1, then cyclization of intermediate 1 was performed by addition of carbon disulfide (CS2) in the presence of anhydrous potassium hydroxide (KOH) in absolute ethanol to afford the parent nucleus, 5-(4aminophenyl)-1,3,4-oxadiazole-2-thione (2), which underwent nucleophilic substitution reaction with three different aromatic amines, using pyridinium trifluoroacetate (PTFA) as a catalyst, to yield three moieties of 1,2,4-triazole derivatives 3, 4 and 5, respectively. They were analyzed utilizing FTIR and 1HNMR spectroscopy, and their physical properties were measured. The target compounds (3, 4, and 5) were investigated for their plausible anti-proliferative effect against Epidermal Growth Factor Receptor (EGFR) drug target using molecular modeling, docking and simulations studies. As per docking results, compound 5 has shown the best inhibitory potential by showing binding energy of -6.38Kcal/mol. Compounds 3 and 4 were shown to be the second and third best with estimated binding free energies of -6.12kcal/mol, and, followed by -6.02kcal/mol, respectively. Several interesting molecular level interactions have been revealed during the molecular dynamic (MD) simulation of compound 5 with EGFR drug target. To validate these insilico predictions, the three compounds were tested on two human cell lines, HeLa, and MCF-7 cancer cell lines, to check their antiproliferative activity. The cytotoxicity for compound 5 in this series was higher with IC50 value measured as 22.83μg/ml and 20.14 μg/ml for HeLa and MCF-7 cell lines, respectively in comparison with compound 3 (IC50 value was 36.71 μg/ml, 35.44 μg/ml), and compound 4, which demonstrated the lowest activity against both cell lines with IC50 value of 47.19 μg/ml, 45.82 μg/ml, HeLa and MCF-7 cell lines respectively. Invitro studies output was in agreement with insilico predictions that compound 5 among the tested compounds has the best potential to act as an anti-proliferative agent.
Synthesis, cytotoxicity, in vitro antiproliferative activity, docking, MD simulations.