title: Production of solid lipid submicron particles for protein delivery using a novel supercritical gas-assisted melting atomization process.
abstract: A supercritical carbon dioxide micronization technique based on gas-assisted melting atomization has been designed to prepare protein-loaded solid lipid submicron particles. The supercritical process was applied to homogeneous dispersions of insulin in lipid mixtures: (1) tristearin, Tween-80, phosphatidylcholine and 5 kDa PEG (1:0.1:0.9:1 and 1:0.1:0.9:2 weight ratio); and (2) tristearin, dioctyl sulfosuccinate and phosphatidylcholine (1:1:0.5 weight ratio). Optimized process conditions yielded dry nonagglomerated powders with high product recovery (70%, w/w). Dynamic light scattering and transmission electron microscopy showed that two size fractions of particles, with 80-120 and 200-400 nm diameters, were produced. In all final products, dimethylsulfoxide used to prepare the insulin/lipid mixture was below 20 ppm. Protein encapsulation efficiency increased up to 80% as the DMSO content in the insulin/lipid mixture increased. Compared to the particles without PEG, the polymer-containing particles dispersed rapidly in water, and the dispersions were more stable under centrifugation as less than 20% of suspended particles precipitated after extensive centrifugation. In vitro, the protein was slowly released from the formulation without PEG, while a burst and faster release were obtained from the formulations containing PEG. Subcutaneous injection to diabetic mice of insulin extracted from the particles showed that the supercritical process did not impair the protein hypoglycemic activity.
authors.affiliation: Department of Pharmaceutical Sciences, University of Padova, Padova, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Salmaso
authors.forename: Nicola
authors.initials: N
authors.lastname: Elvassore
authors.forename: Alberto
authors.initials: A
authors.lastname: Bertucco
authors.forename: Paolo
authors.initials: P
authors.lastname: Caliceti
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000187:drug effects; D002245:Carbon Dioxide / Q000737:chemistry*; D002626:Chemistry, Pharmaceutical; D025924:Chromatography, Supercritical Fluid / Q000295:instrumentation; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000188:drug therapy; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D004143:Dioctyl Sulfosuccinic Acid / Q000737:chemistry; D004337:Drug Carriers; D004339:Drug Compounding; D004355:Drug Stability; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000737:chemistry*; D007279:Injections, Subcutaneous; D007328:Insulin / Q000008:administration & dosage / Q000737:chemistry*; D007700:Kinetics; D008055:Lipids / Q000737:chemistry*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009330:Nebulizers and Vaporizers; D010316:Particle Size; D010713:Phosphatidylcholines / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011136:Polysorbates / Q000737:chemistry; D011208:Powders; D011489:Protein Denaturation; D055550:Protein Stability; D012995:Solubility; D013678:Technology, Pharmaceutical / Q000295:instrumentation / Q000379:methods*; D014280:Triglycerides / Q000737:chemistry
chemical_list: D001786:Blood Glucose; D004337:Drug Carriers; D007004:Hypoglycemic Agents; D007328:Insulin; D008055:Lipids; D010713:Phosphatidylcholines; D011136:Polysorbates; D011208:Powders; D014280:Triglycerides; D004143:Dioctyl Sulfosuccinic Acid; D002245:Carbon Dioxide; D011092:Polyethylene Glycols; C022618:tristearin; D004121:Dimethyl Sulfoxide