Modulatory potential of A novel benzofuran-derived compound against aluminium chloride-induced neurotoxicity and miRNA dysregulation: Biochemical and bio-analytical study

The aim of the current work was to investigate the neuro-modulatory activities of a novel synthesized compound, namely, 3-((3-Acetylphenyl) amino)-1-(benzofuran-2-yl) prop-2-en-1-one (designated as <b>compound IV</b>) in aluminium-chloride (AlCl3)-intoxicated rats. Moreover, for the first time, quantitative analysis of compound-IV in rat plasma was developed using a novel, properly validated Ultra-High-Performance-Liquid Chromatography / Ultra-Violet (UHPLC/UV) method. For the biochemical studies; four-groups were included: negative-control, AlCl₃-intoxicated-rats, intoxicated-rats treated with compound-IV, and reference-donepezil, respectively. Biochemical/molecular assays were conducted; levels of interleukin-6 (IL-6), total antioxidant-capacity (TAC), brain-derived-neurotrophic-factor (BDNF), and total protein content (TP). Differential expressions of miRNA-34a, miRNA-15a, and miRNA-132, were assessed. For the bio-analytical studies; several chromatographic-conditions and extraction-procedures were meticulously optimized. Biochemical results revealed that AlCl₃ (a neurotoxic-agent), enhanced neuroinflammation, oxidative-stress and synaptic-dysfunction; as indicated by increased IL-6 levels, declined both TAC levels and BDNF contents. Moreover, significant dysregulation in miR-34a, -15a, and -132 levels were observed. In contrast, treatment of neuro-intoxicated rats with compound-IV ameliorated all the investigated biomarkers. This novel-benzofuran-derivative exerts its neurotherapeutic-activity by reducing AlCl₃-induced neurotoxicity and mitigating oxidative-stress, neuroinflammation, and synaptic-dysfunction through regulating all miRNA levels. The developed and validated analytical-method ensured that best quantitative separation of compound-IV was achieved using Symmetry-C18-column, with mobile phase consisting of acetonitrile: H₂O (50: 50), UV detection at λmax 390-nm, 1 mL/min flow-rate and 3.4 min retention time. The proposed method provides excellent specificity and linearity over concentration range of 1-100 μg/mL; hence, it could serve as a perquisite-step for further investigation of bioavailability (BA) and pharmacokinetics (PKs) of this compound.