Integrated Synthetic, Pharmacological and In-Silico Evaluation of Novel Heterocyclic Compounds Targeting Cancer Pathways

Abstract

The present investigation was undertaken to design, synthesize, characterize and biologically evaluate novel heterocyclic derivatives with the objective of discovering new small-molecule leads possessing analgesic, anti-inflammatory and anticancer potential, supported by molecular docking-based mechanistic insight. Major heterocyclic systems were explored: 7-Azaindole / 7-azaisatin–derived semicarbazide hybrids (VIIa–l and selected analogues) for anticancer screening. All synthesized compounds were structurally confirmed through IR, ¹H-NMR, ¹³C-NMR, LC–MS and elemental analysis, with melting-point and TLC supporting purity. The synthetic strategies used versatile heterocyclic chemistry enabling substitution-dependent structure–activity evaluation. Acute oral toxicity studies demonstrated wide safety margins, with no mortality up to high dose levels in rodents, suggesting good tolerability. Pharmacological Findings For the azaindole / azaisatin semicarbazide derivatives: In vivo anticancer screening using Ehrlich Ascites Carcinoma demonstrated significant tumor-growth suppression, improved survival indices, and reduced viable cell counts for selected compounds. MTT cytotoxicity studies confirmed dose-dependent antiproliferative activity against HeLa and HCT-15 cell lines, with micromolar-level IC₅₀ values for the most active molecules (e.g., XXV and XXIIIb). Docking against DHFR and COX-2 provided mechanistic evidence of favorable receptor complementarity. Collectively, the findings validate the medicinal-chemistry hypothesis that heterocyclic hybridization and rational substitution drive potency, binding affinity, and biological response across multiple pharmacological models.