Exploring the therapeutic potential of benzodioxane carboxylic acid-based hydrazones: Structural, computational, and biological insights

The present study investigates a series of benzodioxane carboxylic acid-based hydrazones for their structural, physicochemical, pharmacokinetic, and biological properties. A multi-step synthesis approach was employed to prepare novel derivatives (Compounds 4–7), which were characterized using UV-Visible spectroscopy, Fourier transform infrared (FTIR), 1H nuclear magnetic resonance (NMR), 13C NMR, and electron ionization-mass spectroscopy (EI-MS) to confirm their molecular structures. The compounds were evaluated through molecular docking, molecular dynamics (MD) simulations, and various in vitro assays. The synthesized hydrazones demonstrated diverse biological properties, including antibacterial, antifungal, anticancer, antioxidant, and enzyme inhibition activities. Computational studies revealed strong binding interactions and stability for Compounds 5 and 7, which contain methoxy and sulfur-containing thiophene groups. These substituents facilitated hydrogen bonding, π-π stacking, and hydrophobic interactions within receptor active sites. The physicochemical evaluation confirmed the drug-like properties of all compounds, which adhere to Lipinski's Rule of Five, suggesting good oral bioavailability. Additionally, electronic characteristics, including dipole moments, energy gaps, and molecular electrostatic potential (MEP), provided insights into the structural influences on bioactivity. The in vitro biological activities aligned with in silico predictions, with compound 5 exhibiting potent inhibition of acetylcholinesterase (IC₅₀ = 1.228 ± 1.63 µg/mL), β-glucosidase (IC₅₀ = 0.37 ± 3.06 µg/mL), and peroxidase (IC₅₀ = 2.009 ± 3.19 µg/mL), along with significant anticancer activity (50.17% inhibition against HeLa cells). Compound 7, featuring a sulfur thiophene ring, exhibited strong antioxidant (IC₅₀ = 17.16 ± 2.641 µg/mL) and anticancer (37.11% inhibition against PC3 cells) activities, as well as enzyme inhibition against α-amylase (IC₅₀ = 2.81 ± 8.69 µg/mL), tyrosinase (IC₅₀ = 0.70 ± 2.30 µg/mL), and lipase. The antimicrobial activity of Compound 5 was particularly notable against Escherichia coli and Bacillus subtilis, whereas compound 6 exhibited limited antimicrobial effects. SAR analysis highlighted the critical role of functional groups, such as methoxy and sulfur-containing thiophene rings, in enhancing biological activities. These findings underscore the importance of specific substituents, such as methoxy, thiophene, and trifluoromethyl groups, in modulating bioactivity. Compounds 5 and 7 emerged as promising candidates for therapeutic applications, particularly in anticancer, neurodegenerative, and metabolic disorders, while compounds 4 and 6 exhibited moderate biological activity. Furthermore, the physicochemical properties confirmed good oral bioavailability, supporting further development for drug design.