Synthesis, cytotoxic evaluation, DFT analysis, molecular dynamics simulation and ADME studies of novel 1-(4-(substituted-1-(p-tolyl)-3-(trifluoromethyl)-1H-pyrazol-5-yl)-4-methylpiperazine derivatives

Abstract

This study presents the rational design, synthesis, and pharmacological evaluation of a novel series of fifteen 1- (4-(substituted)-1-(p-tolyl)-3-(trifluoromethyl)-1H-pyrazol-5-yl)-4-methylpiperazine derivatives (7a-7o), developed by integrating a trifluoromethylated pyrazole core with a bioreducible arylpiperazine moiety to target nonsmall cell lung cancer (NSCLC). All compounds were structurally confirmed using advanced spectroscopic techniques, and in vitro cytotoxic activity was assessed against the A-549 cell line via the 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay. Among them, 3-methoxy-N-((5-(4-methylpiperazin-1-yl)-1- (p-tolyl)-3-(trifluoromethyl)-1H-pyrazol-4-yl)methyl)aniline (7m) and 4-isopropyl-N-((5-(4-methylpiperazin-1- yl)-1-(p-tolyl)-3-(trifluoromethyl)-1H-pyrazol-4-yl)methyl)aniline (7o) exhibited the most potent antiproliferative effects (IC₅₀ ≈ 69 μg/mL). Molecular docking and molecular dynamics studies supported these findings, with compound 7m showing a strong binding affinity (-8.114 kcal/mol) toward the target protein (PDB ID: 1M17) through a key hydrogen bond with CYS773 and extensive hydrophobic interactions, and lower root mean square deviation (RMSD) values, respectively. Absorption, distribution, metabolism, and excretion (ADME) predictions confirmed favorable pharmacokinetic characteristics, including compliance with Lipinski’s criteria and 100% predicted oral absorption. Collectively, these results establish the trifluoromethylated pyrazolepiperazine scaffold as a promising lead framework for the development of next-generation NSCLC therapeutics. Density functional theory (DFT) computations were carried out to discover the electrical structure and molecular characteristics of the system. DFT achieves an efficient balance between computing cost and precision by representing the ground-state energy as a functional of electron density.