Integration of Design of Experiments for the Development and Evaluation of Sulfasalazine Loaded Hybrid Albumin Chitosan Based Polymeric Nanocarriers.

Sulfasalazine is a sepiapterin reductase and System x - c inhibitor with anti-neuroblastoma activity. The objective was to develop cellular redox glutathione stimuli-responsive Sulfasalazine-loaded albumin-chitosan hybrid nanocarriers with a target profile of < 100 nm, polydispersity index (PDI) of < 0.3, charge of 3 to 10 mV, and drug load of > 8% w/w. Taguchi orthogonal array experimental design was utilized in the formulation of cellular redox glutathione stimuli-responsive albumin-chitosan-based hybrid nanocarriers using the nano-precipitation method. Chitosan was coated on the Albumin nanocarriers, followed by PEGylation. Nanocarriers were characterized in terms of size, zeta potential, polydispersity index (PDI), entrapment efficiency, and drug release. The uptake of chitosan-coated sulfasalazine-loaded albumin nanocarriers was investigated using SK-N-Be(2)c neuroblastoma cells. Taguchi orthogonal array results revealed that the acetone concentration and organic phase to aqueous phase ratio were the significant variables in formulating stable monodispersed nanocarriers with mean particle size of 80.55 ± 3.30 nm, polydispersity index (PDI) of 0.120 ± 0.037, zeta potential of 6.60 ± 1.95 mV, entrapment efficiency of 96.42 ± 1.38%, and 10% w/w drug load. The concentration of acetone had a significant impact on the size and PDI of Albumin nanocarriers. The cellular redox glutathione stimuli-responsive nanocarriers exhibited an extended drug release of 30 ± 1% at 10 mM, an intracellular concentration, and 25 ± 1% at 20 mM glutathione, an extracellular systemic concentration in PBS-based release media. PEGylation of nanocarriers confers stability in ionic environments. Sulfasalazine-loaded albumin-chitosan nanocarriers showed significantly higher SK-N-BE (2)c cellular uptake than Albumin nanocarriers (p < 0.001). The Taguchi orthogonal array design was successfully applied in the development of cellular redox glutathione stimuli-responsive sulfasalazine-loaded extended-release albumin-chitosan hybrid nanocarriers, which met the target profile of < 100 nm, a polydispersity index (PDI) of < 0.3, a charge of 3 to 10 mV, and a drug load of > 8% w/w against neuroblastoma.