Pyrazoles and diazepines are important heterocyclic compounds with significant biological and pharmacological potential. In this study, a series of newly synthesized pyrazole and diazepine derivatives was prepared using conventional organic methods from chalcone intermediates derived from 4-aminoacetophenone, which reacted with various substituted anilines. FT-IR and H NMR spectroscopy confirmed, and molecular docking studies revealed that both the pyrazole and diazepine derivatives exhibited favourable binding affinities for glucosamine-6-phosphate synthase by forming multiple stabilizing interactions within the enzyme's active site. These findings suggest a promising potential for these compounds to act as enzyme inhibitors. Biological evaluation showed that some derivatives had considerable antibacterial activity, with compound 5 being especially effective against E. coli and S. aureus; its efficacy increased with concentration. Additionally, derivative 5 demonstrated notable cytotoxicity against the MCF-7 breast cancer cell line, reducing cell viability in a clear dose-dependent manner, indicating its potential as an anticancer candidate. Thus, the combined approach of synthesis, spectral and spectroscopic characterization, molecular docking, and biological testing provides a deeper understanding of the structure-activity relationships in these new derivatives. The results highlight the potential of pyrazole and diazepine scaffolds as leads for developing promising antibacterial and anticancer agents, warranting further optimization and biological evaluation.