Structure–Activity Relationships of Ferrocenyl Acetylaniline Derivatives as Dual Inhibitors of Carbohydrate-Hydrolysing Enzymes and Inflammatory Processes: Combined Experimental and Computational Insights

Abstract

This thesis reports the design, synthesis, physicochemical characterization, and biological evaluation of three ferrocenyl acetylaniline derivatives, namely FcMe2Ac, FcMe3Ac, and FcMe4Ac, with the aim of exploring their potential as dual antidiabetic and anti-inflammatory agents. The compounds were synthesized through a nucleophilic substitution approach, affording moderate to high yields (48–80%). Their structures were confirmed by FT-IR, UV–Visible, and one- and two-dimensional NMR spectroscopy, which verified the successful incorporation of both ferrocenyl and acetylaniline moieties. Electrochemical studies using cyclic voltammetry revealed quasi-reversible Fe(II)/Fe(III) redox behavior for all derivatives, highlighting the influence of substitution patterns on electron-transfer properties. Biological assays demonstrated significant inhibition of the carbohydrate-hydrolyzing enzymes α-amylase and α-glucosidase, with IC₅₀ values ranging from 8.06 to 10.39 µg·mL⁻¹. Among the tested compounds, FcMe4Ac exhibited the strongest antidiabetic activity, outperforming the reference drug acarbose. The derivatives also displayed notable anti-inflammatory activity through inhibition of bovine serum albumin denaturation, with IC₅₀ values between 4.22 and 7.29 µg·mL⁻¹, where FcMe4Ac again showed superior efficacy compared with diclofenac. Density functional theory (DFT) calculations provided insight into the electronic properties of the molecules, indicating moderate stability and favorable reactivity. Molecular docking studies revealed strong binding affinities toward α-amylase and α-glucosidase, supported by hydrophobic interactions, hydrogen bonding, and π–π contacts within the active sites. Molecular dynamics simulations confirmed the stability of the ligand–enzyme complexes over time. Furthermore, ADMET predictions suggested favorable pharmacokinetic and safety profiles, including high gastrointestinal absorption, balanced lipophilicity, and low predicted toxicity. Overall, the results identify FcMe4Ac as a promising lead compound for the development of multifunctional agents targeting diabetes and inflammation.