Quality by Design-Based Development and Optimization of Berberine Hydrochloride-Loaded Nanoliposomes for Enhanced Antidiabetic Activity

Abstract

Berberine hydrochloride (BBR-HCl) is a plant-derived isoquinoline alkaloid with multiple metabolic actions relevant to type 2 diabetes mellitus, but its pharmaceutical performance after oral administration is limited by poor membrane permeability, intestinal efflux, extensive presystemic metabolism and low bioavailability. The present study was designed to develop and optimize BBR-HCl-loaded nanoliposomes using a Quality by Design (QbD) framework and to evaluate their antidiabetic performance exclusively through in-vitro and cell-based experiments. A Quality Target Product Profile was defined and vesicle size, polydispersity index (PDI), zeta potential, drug content, entrapment efficiency and release behavior were identified as critical quality attributes. Risk assessment identified phosphatidylcholine concentration, cholesterol concentration and sonication time as critical variables. A three-factor, three-level Box-Behnken design was applied to optimize vesicle size, PDI and entrapment efficiency. Nanoliposomes were prepared by thin-film hydration followed by probe sonication and were characterized by dynamic light scattering, electrophoretic mobility, transmission electron microscopy, Fourier-transform infrared spectroscopy and differential scanning calorimetry. In-vitro release, alpha-amylase inhibition, alpha-glucosidase inhibition, cytocompatibility and glucose uptake in insulin-resistant L6 myotubes were investigated. In the illustrative dataset, the optimized formulation showed a mean vesicle size of 157.2 +/- 2.8 nm, PDI of 0.204 +/- 0.008, zeta potential of -24.6 +/- 1.7 mV and entrapment efficiency of 89.1 +/- 1.0%. The formulation provided prolonged release over 24 h and produced stronger apparent inhibition of alpha-amylase and alpha-glucosidase than free BBR-HCl. It also enhanced glucose uptake in insulin-resistant L6 myotubes while maintaining acceptable cellular viability. QbD-based optimization therefore offers a rational platform for developing reproducible berberine nanoliposomes and supports further non-animal permeability, mechanistic, safety and clinical translation studies.