Published November 18, 2025
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Solid lipid nanoparticles loaded with a novel indole-based multitarget ligand: preparation, characterization, and neuroprotective evaluation in an H₂O₂-induced SH-SY5Y cell model
Authors/Creators
- 1. Medical University of Sofia, Sofia, Bulgaria
Description
A newly synthesized indole-based N-benzylpiperidine derivative (3a), acting as a multitarget-directed ligand with acetylcholinesterase inhibitory activity, was encapsulated into solid lipid nanoparticles (SLNs) to improve neuroprotective potential and enable intranasal brain delivery in future in vivo studies. Lipid prescreening was performed, and a spectrophotometric method for the quantification of 3a was developed and validated. SLNs were prepared using the nano-template engineering technique and evaluated for particle size, zeta potential, drug entrapment efficiency, in vitro release, and stability. The optimized formulation (3a-SLNs) was further characterized by transmission electron microscopy (TEM), X-ray powder diffraction (XRD), and differential scanning calorimetry (DSC). The nanoparticles exhibited favorable physicochemical properties, releasing 52% of the drug over 24 h. Permeability studies indicated improved cellular uptake of 3a upon SLN encapsulation. The formulation remained stable for 6 months under refrigerated (4 ± 2 °C) and room temperature (25 ± 2 °C; 60 ± 5% RH) conditions. The neuroprotective potential of 3a-SLNs was evaluated in an H₂O₂-induced oxidative stress model in SH-SY5Y neuronal cells using the MTT assay. Cells were pretreated with free 3a (0.1–50 µM) for 90 min before H₂O₂ exposure (1 mM). Hydrogen peroxide induces oxidative damage via reactive hydroxyl radicals, affecting cellular lipids, proteins, and DNA. 3a-SLNs exhibited superior protective effects compared with free 3a, indicating that nanoparticle encapsulation enhances intracellular availability and antioxidant defense. These findings highlight SLN-encapsulated 3a as a promising intranasal nanocarrier system for Alzheimer's disease therapy, which will be further investigated in in vivo models. By improving brain delivery potential and reducing oxidative stress–induced neuronal injury, 3a-SLNs offer a strategy to overcome pharmacokinetic limitations of conventional compounds and enhance neuroprotective efficacy.
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