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

## TL;DR

A new benzofuran compound reduces aluminum chloride-induced brain damage and miRNA imbalance in rats, with a new method to measure its levels in blood.

## Contribution

A novel benzofuran-derived compound and a validated UHPLC/UV method for its quantification in rat plasma are introduced.

## Key findings

- AlCl3 caused neuroinflammation, oxidative stress, and miRNA dysregulation in rats.
- Compound IV reversed AlCl3-induced biomarker changes and miRNA levels.
- The UHPLC/UV method provided accurate quantification of compound IV in plasma.

## Abstract

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 compound IV) 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.

•Aluminium-chloride (AlCl3) enhances neuroinflammation and synaptic dysfunction.•AlCl3 stimulated miRNA dysregulation including miRNA-34a, miRNA-15a, and miRNA-132.•Compound (IV) demonstrated therapeutic potential against AlCl3-induced neurotoxicity.•The UHPLC/uv method offers a precise, low-cost technique to quantify Compound (IV)•The method may enable future estimation of primary and secondary PK parameters

Aluminium-chloride (AlCl3) enhances neuroinflammation and synaptic dysfunction.

AlCl3 stimulated miRNA dysregulation including miRNA-34a, miRNA-15a, and miRNA-132.

Compound (IV) demonstrated therapeutic potential against AlCl3-induced neurotoxicity.

The UHPLC/uv method offers a precise, low-cost technique to quantify Compound (IV)

The method may enable future estimation of primary and secondary PK parameters