The processing of nanoparticles containing protein for suspension in hydrofluoroalkane propellants.

Nanoparticles delivered from pressurized metered dose inhalers (pMDIs) potentially offer a means of efficiently delivering proteins to the lung. Nanoparticles containing the model protein lysozyme have been produced using microemulsion and nanoprecipitation methods. Freeze-drying water in oil emulsions, with chloroform as the organic solvent, followed by washing of excess surfactant (lecithin) led to the production of lactose nanoparticles having approximately 300 nm mean size. Substitution of lactose with lysozyme led to a significant increase in the mean size of nanoparticles (645-750 nm). This may have been due to the surface activity of lysozyme which altered the emulsification properties. The retained biological activity of lysozyme increased with increased lactose concentration in the formulation, and approximately 99% biological activity was retained when 20% (w/w) lactose was used. Ethanol used in the formulation in place of chloroform changed the production process from emulsification to nanoprecipitation. A monodisperse system (mean size approximately 275 nm, polydispersity index approximately 0.1) of spherical nanoparticles containing 80% (w/w) bioactive lysozyme (retained activity 99%) was generated. The nanoparticles washed with ethanol containing DPPC, oleic acid or Span 85 (2%, w/v) could readily be dispersed in HFA 134a without further processing, and a stable suspension was formed. Lysozyme remained stable (retained biological activity 98%) even after the nanoparticles were suspended in HFA 134a. This indicates the potential of nanoparticles for delivery of proteins from HFA-based pressurized metered dose inhaler formulations.

The anaerobic dechlorination of trichlorofluoromethane by rat liver preparations in vitro.

Incubation of trichlorofluoromethane with a liver microsomal fraction and an NADPH generating system under anaerobic conditions produced a metabolite dichlorofluoromethane, characterised by gas chromatography and mass spectrometry. The metabolic reaction was carried out by liver microsomes from the mouse, rabbit, hamster and rat and was increased by phenobarbitone pre-treatment. The formation of dichlorofluoromethane in vitro was enhanced by the addition of FMN, but partially inhibited by the presence of air, oxygen, SK&F 525-A, metyrapone and carbon tetrachloride and totally inhibited by carbon monoxide. The consumption of NADPH in the reaction was greater than could be accounted for by the production of dichlorofluoromethane indicating the possible formation of other metabolic products. It is suggested that trichlorofluoromethane interacts with the reduced form of cytochrome P-450 at the oxygen binding site and a possible mechanism for its subsequent reductive dechlorination is proposed.

Fluorocarbon aerosol propellants XI: Pharmacokinetics of dichlorodifluoromethane in dogs following single and multiple dosing.

A three-compartment open model was proposed for the disposition of dichlorodifluoromethane in dogs with average half-lives of 1.47, 7.95, and 58.5 min for the three disposition phases. This proposal is contrary to several studies that monitored blood levels for a shorter period. An analysis of the tissue compartment distribution following a single dose showed that about 1.5 hr was required to achieve pseudodistribution equilibration, following which more than 90% of the dose remaining in the body was retained in the tissue compartments. The pulmonary clearance and volumes of distribution were calculated considering the first-pass effect through the lungs. The volume of distribution after reaching pseudodistribution equilibrium was approximately 10 times the body weight in terms of the blood concentration, and about 68% of the propellant was cleared from the blood passing through the lungs in each cycle. Disposition of the propellant followed dose-independent kinetics after multiple dosing. No volatile metabolites were detected in the blood using GC.

Study on the binding of trichloromonofluoromethane by bovine serum albumin using a fluorescent probe technique.

Protein binding of the aerosol propellant, trichloromonofluoromethane, was studied in bovine serum albumin (BSA) solutions using the fluorescent probe technique. The propellant displaced the probe, 8-anilino-1-naphthalenesulfonate, from its binding sites and reduced the fluorescence intensity. The binding association constants, the number of binding sites, and the competitive nature of the binding interaction were investigated. The hydrophobic nature of the binding and the implication of binding displacement interactions between the fluorocarbon and plasma-protein-bound drugs were also discussed. The fatty acid impurities present in the commercial BSA were found to have no effect on the protein binding of the propellant.

Fluorocarbon aerosol propellants X: pharmacokinetics of dichlorotetrafluoroethane in dogs.

An intravenous dosage form of dichlorotetrafluoroethane, a common fluorocarbon aerosol propellant, was formulated in polyethylene glycol 400 for single dosing to unanesthetized dogs. A three-compartment open model was proposed for the disposition of this compound in dogs, with average half-lives of 1.3, 9.6, and 50.8 min for the three disposition phases. An analysis of tissue compartment distribution following a single dose showed that it took about 2 hr to achieve pseudo-distribution equilibration, following which more than 90% of the propellant remaining in the body was retained in the tissue compartments. Pulmonary clearance and volumes of distribution were calculated considering the first-pass effect through the lungs. The volume of distribution was approximately 10 times the body weight in terms of blood concentration, and about 84% of the propellant was cleared from the blood passing through the lungs in each cycle.

Fluorocarbon aerosol propellants IV: pharmacokinetics of trichloromonofluoromethane following single and multiple dosing in dogs.

An intravenous dosage form of trichloromonofluoromethane, a common aerosol propellant, was formulated in polyethylene glycol 400 for single and multiple dosing to unanesthetized dogs. A three-compartment open model was proposed for disposition of this compound in dogs with average half-lives of 3.2, 16, and 93 min for three disposition phases. This finding is contrary to several reports where blood levels were monitored for shorter periods. A computer analysis of tissue compartment distribution following a single dose showed that about 2 hr was required to achieve pseudodistribution equilibration, following which more than 90% of the dose remaining in the body was retained in tissue compartments. Pulmonary clearance and volumes of distribution were calculated considering first-pass effect through the lung. The volume of distribution was approximately six times the body weight in terms of blood concentrations, and about 30% of the propellant was cleared from blood passing through the lung in each cycle. Disposition of propellant followed dose-independent kinetics after multiple dosing, and accumulation in tissues continued for a much longer period, resulting in high tissue compartment levels.