Interaction of poly(butylcyanoacrylate) nanoparticles with the blood-brain barrier in vivo and in vitro.

Poly(butylcyanoacrylate) nanoparticles were produced by emulsion polymerisation and used either uncoated or overcoated with polysorbate 80 (Tween 80). [3H]-dalargin bound to nanoparticles overcoated with polysorbate 80 or in the form of saline solution was injected into mice and the brain concentrations of radioactivity determined. Statistically significant, three-fold higher brain concentrations with the nanoparticle preparations were obtained after 45 minutes, the time of greatest pharmacological response assessed as analgesia in previous experiments. In addition the brain inulin spaces in rats and the uptake of fluoresceine isothiocyanate labelled nanoparticles in immortalised rat cerebral endothelial cells, (RBE4) were measured. The inulin spaces after i.v. injection of polysorbate 80-coated nanoparticles were significantly increased by 1% compared to controls. This is interpreted as indicating that there is no large scale opening of the tight junctions of the brain endothelium by the polysorbate 80-coated nanoparticles. In in vitro experiments endocytic uptake of fluorescent nanoparticles by RBE4 cells was only observed after polysorbate 80-overcoating, not with uncoated particles. These results further support the hypothesis that the mechanism of blood-brain barrier transport of drugs by polysorbate 80-coated nanoparticles is one of endocytosis followed by possible transcytosis. The experiments were conducted in several laboratories as part of an EEC/INTAS collaborative program. For various procedural and regulatory reasons this necessitated the use of both rats and mice as experimental animals. The brain endothelial cell line used for the in vitro studies is the rat RBE4.

Dissolution testing as a prognostic tool for oral drug absorption: dissolution behavior of glibenclamide.

PURPOSE: The dissolution behavior of two commercially available glibenclamide formulations was tested in various media. The aim of the study was to investigate whether the use of biorelevant dissolution media (BDM) would be advantageous over the use of standard media for predicting the in vivo performance of the two formulations. METHODS: The dissolution tests were performed using USP 23 apparatus 2. Conventional buffers and USP media were compared with two BDM containing different amounts of lecithin and sodium taurocholate. RESULTS: The dissolution of two drug powders was highly dependent on wetting, particle size, pH, and the composition of the medium used. In addition, the dissolution behavior of the two glibenclamide formulations showed differences in all media tested. The dissolution results of the two formulations were compared with those from an in vivo bioequivalence study undertaken by the central quality control laboratory of the German pharmacists (ZL). The bioequivalence criterion set by the ZL requires more than 80% drug release within 10 minutes. Results in FaSSIF, one of the BDMs, met the ZL criterion and this medium was also able to discriminate between the two formulations. This was not the case for the other media tested. CONCLUSIONS: The study indicates that BDM are better able to discriminate between glibenclamide formulations than standard dissolution media.

Modern bioavailability, bioequivalence and biopharmaceutics classification system. New scientific approaches to international regulatory standards.

In the last decade, the regulatory bioequivalence (BE) requirements of drug products have undergone major changes. The introduction of the biopharmaceutics drug classification system (BCS) into the guidelines of the Food and Drug Administration (FDA) is a major step forward to classify the biopharmaceutical properties of drugs and drug products. Based on mechanistic approaches to the drug absorption and dissolution processes, the BCS enables the regulatory bodies to simplify and improve the drug approval process. The knowledge of the BCS characteristics of a drug in a formulation can also be utilized by the formulation scientist to develop a more optimized dosage form based on fundamental mechanistic, rather than empirical, information. This report gives a brief overview of the BCS and its implications.

Influence of the surfactant concentration on the body distribution of nanoparticles.

The rapid reticuloendothelial system (RES) uptake of nanoparticles after i.v. injection, especially by the liver, can be reduced and the body distribution can be altered by coating them with non-ionic surfactants. In the present work 2-14C-poly(methyl methacrylate) nanoparticles were coated with poloxamine 908 and polysorbate 80, and the influence of different surfactant concentrations on the body distribution was investigated. These surfactants were chosen because earlier studies showed that poloxamine 908 was very effective in decreasing the liver uptake and keeping the nanoparticles in circulation, whereas polysorbate 80 was the most effective surfactant to direct the particles to organs that do not belong to the RES. Above nanoparticles were injected i.v. to rats and the animals were sacrificed after 30 min. Below a surfactant concentration of 0.1% the nanoparticle preparations behaved like uncoated particles. At a 0.1% concentration a very sudden and significant change in the body distribution occurred with poloxamine 908. The liver concentration decreased from about 75% of the dose to 13% and stayed at this level at higher surfactant concentrations. This decrease was combined with a similar sudden complementary increase in blood and other organ and tissue concentrations. With polysorbate 80 the decrease in liver concentration and increase in the blood and the other organ levels was gradual and became important only above 0.5% surfactant concentration. The results indicate that the type of interaction and the strength of the adsorptive binding to the nanoparticles are different with different surfactants. This in turn leads to different body distribution patterns after i.v. injection of surfactant coated nanoparticles.

Body distribution of azidothymidine bound to hexyl-cyanoacrylate nanoparticles after i.v. injection to rats.

Cells of the reticuloendothelial system (RES e.g. macrophages) play an important role in the immunopathogenesis of AIDS. The objective of the present study was to investigate the possibility of specifically targeting antiviral drugs such as azidothymidine (AZT) to macrophages using nanoparticles as colloidal drug carriers. In a first series of experiments the body distribution of 14C-labelled AZT bound to nanoparticles and a similarly prepared control solution with unbound AZT were studied in rats after intravenous injection. In a second series of experiments polysorbate 80-coated nanoparticles and a solution of AZT in saline were tested. 14C-labelled AZT was bound to nanoparticles using the surfactant bis(2-ethylhexyl) sulphosuccinate sodium (DOSS). The radioactivity in several organs, including those containing large numbers of macrophages, was measured after intravenous injection of the AZT-nanoparticles and the AZT-control solutions. AZT concentrations were up to 18 times higher in organs belonging to the RES if the drug was bound to nanoparticles compared with unbound AZT. These results demonstrate that nanoparticles are a potential drug targeting system for anti-AIDS drugs. The increase in drug concentration at the sites containing abundant macrophages may allow a reduction in dosage to reduce systemic toxicity.

Evaluation of various dissolution media for predicting in vivo performance of class I and II drugs.

PURPOSE: In this paper we seek to verify the differences in dissolution behavior between class I and class II drugs and to evaluate the suitability of two new physiologically based media, of Simulated Gastric Fluid (SGF) and of milk for their ability to forecast trends in the in vivo performance of class II compounds and their formulations. METHODS: Dissolution behavior of two class I drugs, i.e. acetaminophen and metoprolol, and of three class II drugs, i.e. danazol, mefenamic acid and ketoconazole, was studied with USP Apparatus 2 in water, SGF, milk, Simulated Intestinal Fluid without pancreatin (SIFsp) and in two media simulating the small intestinal contents in the fed (FeSSIF) and fasted (FaSSIF) states, respectively. RESULTS: Class I powders dissolved rapidly in all media tested. Acetaminophen dissolution in milk was slow from one tablet formulation, in all other cases dissolution was more than 85% complete in 15 minutes. The dissolution rate of metoprolol was shown to be dependent on formulation and manufacturing method, and one of the three tablet formulations did not meet compendial specifications (80%/30 minutes). Dissolution behavior of class II drugs was greatly affected by choice of medium. Dissolution from a capsule formulation of danazol proved to be dependent on the concentration of solubilizing agents, with a the 30-fold increase in percentage dissolved within 90 minutes upon changing from aqueous media without surfactants to FaSSIF. Use of FeSSIF or milk as the dissolution medium resulted in an even greater increase in percentage dissolved, 100 and 180-fold respectively. Dissolution of the weak acid mefenamic acid from a capsule formulation is dependent on both pH and bile salt concentration, which leads to an offset between increased bile salt concentration and lower pH in the fed state compared to the fasted state medium. The weak base ketoconazole showed complete dissolution from a tablet formulation in Simulated Gastric Fluid without pepsin (SGFsp) within 30 minutes, 70% dissolution in 2 hours under fed state simulated upper jejunal conditions but only 6% dissolution in 2 hours under fasted state conditions. CONCLUSIONS: As predicted, dissolution of class II drugs proved to be in general much more dependent on the medium than class I drugs. With the array of compendial and physiological media available, it should be possible to design a suitable set of tests to predict the in vivo dissolution of both class I and II drugs from immediate release formulations.

Improved body distribution of 14C-labelled AZT bound to nanoparticles in rats determined by radioluminography.

The objective of the present study is to visualize differences in the body distribution between radiolabelled AZT bound to nanoparticles and a control solution. Polyhexylcyanoacrylate nanoparticles were manufactured by emulsion polymerization in the presence of AZT and an ionic emulsifier, bis(2-ethylhexyl) sulfosuccinate sodium. The AZT-control solution was equally prepared, but contained no monomer. The two preparations were administered either by i.v. injection or perorally by gavage. After determined time points the animals were sacrificed using carbon dioxide. The cadavers were shock-frozen in cellulose gel and cut into slices using a cryomicrotome. The tissue cross sections were fixed on an adhesive tape and then were freeze dried. The quantification of the radioactive AZT in the different organs and tissues was performed by radioluminography, and the images were generated on a computer. After i.v. injection of AZT-nanoparticles, a high amount of the AZT label was found in the organs belonging to the reticuloendothelial system. In these organs the radioactivity was inhomogeneously distributed showing that the uptake of the particle-associated radioactivity depended on the type of the cells located in the organs and was consistent with uptake by macrophages. The highest radioactivities were found in the GI-tract and in the liver. A difference in the elimination pathway between AZT-control solution and AZT bound to nanoparticles also was visible on the images. Similar results were obtained after oral administration. Of course, with the latter route a larger portion of AZT remained in the GI-tract especially after administration of nanoparticle-bound drug. These results confirmed those obtained by a classically performed quantitative whole body distribution study using liquid scintillation. This demonstrates that radioluminography is a useful method to study the organ distribution of drugs bound to nanoparticles.

Macrophage targeting of azidothymidine: a promising strategy for AIDS therapy.

Macrophages play an important role in the immunopathogenesis of AIDS. The objective of this study was to investigate the possibility of specific targeting of antivirals such as azidothymidine (AZT) to macrophages, using nanoparticles as a colloidal drug carrier. The body distribution of AZT bound to nanoparticles and as a control solution was studied in rats after intravenous and peroral administration. 14C-Labeled AZT was bound to nanoparticles in the presence of bis(2-ethylhexyl)sulfosuccinate sodium. The radioactivity was measured in different organs including those containing large numbers of macrophages. After intravenous injection, the concentrations of AZT were up to 18 times higher in organs belonging to the reticuloendothelial system (RES) when the drug was bound to nanoparticles than after injection of an aqueous AZT solution. Likewise, after oral administration the nanoparticle formulation delivered AZT more efficiently to the RES than the aqueous solution. In addition, the blood concentration was significantly higher after oral administration of nanoparticles. These results demonstrate that nanoparticles are a promising drug-targeting system for AZT to the RES organs. The increase in drug availability at the sites containing abundant macrophages may allow a reduction in dosage to avoid systemic toxicity.