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Polymyxins are a group of antibiotics with a common structure of a cyclic peptide with a long hydrophobic tail. Polymyxin B sulphate (PLX) has cationic charge, which is an obstacle for the efficient loading into Solid Lipid Nanoparticles (SLN). In the present paper, we describe an innovative method to load PLX into SLN to achieve the sustained release of the drug. PLX was firstly cross-linked with sodium alginate (SA) at different ratios (1:1, 1:2 and 1:3 SA/PLX), and loaded into SLN produced by high pressure homogenization (HPH). Optimized SLN were produced applying 500 bar pressure and 5 homogenization cycles. The best results were obtained with SA/PLX (1:1), recording 99.08 ± 1.2% for the association efficiency of the drug with SA, 0.99 ± 10 g for the loading capacity and 212.07 ± 5.84% degree of swelling. The rheological profile of aqueous SA solution followed the typical behaviour of concentrated …