James M. Gallo

A Simple Rheological Method for the in Vitro Assessment of Mucin-Polymer Bioadhesive Bond Strength

A simple viscometric method was used to quantify mucin-polymer bioadhesive bond strength. Viscosities of 15% (w/v) porcine gastric mucin dispersions in 0.1 N HC1 (pH 1) or 0.1 N acetate buffer (pH 5.5) were measured with a Brookfield viscometer in the absence (ηm) or presence (ηt) of selected neutral, anionic, and cationic polymers (0.1–2.5%, w/v). Viscosity components of bioadhesion (1%) were calculated from the equation, ηt = ηm + ηp + ηb, where ηp is the viscosity of corresponding pure polymer solution as measured by an Ostwald viscometer. The forces of bioadhesion (F) were calculated from the equation, F = ηbσ, where σ is the rate of shear/sec. ηbs and Fs for polyelectrolytes, e.g., polyacrylic acid, cationic gelatin, and chitosan were always higher in acetate buffer than in HC1. Validity of the technique and the effect of ionic charge, polymer conformation, and rate of shear on ηb and F are discussed, as is a comparison of this method to other methods for evaluating bioadhesive materials.

Optimized Formulation of Magnetic Chitosan Microspheres Containing the Anticancer Agent, Oxantrazole

A combined emulsion/polymer cross-linking/solvent evaporation technique was used to prepare magnetic chitosan microspheres (MCM) containing the anticancer drug, oxantrazole. A central composite experimental design was used to simultaneously evaluate a variety of formulation factors on a number of response variables, such as the percentage of oxantrazole entrapped in the MCM. In association with the study design, statistical optimization procedures indicated the factors that significantly influence MCM preparation and what levels of the factors are needed to produce optimum MCM. Entrapment of anticancer agents into biodegradable microspheres is difficult because of low aqueous drug solubility and porosity of the particles. The latter effect was circumvented by a chitosan cross-linking step that resulted in ∼3% (w/w) oxantrazole entrapment in the MCM via the optimization procedures. The combined formulation and statistical optimization strategy provide a basis to develop other microparticulate systems and led to a dosage form that can be used for future in vivo investigations.

Targeting Anticancer Drugs to the Brain. I: Enhanced Brain Delivery of Oxantrazole following Administration in Magnetic Cationic Microspheres

A magnetic cationic microsphere delivery system, prepared from the polysaccharide chitosan and containing oxantrazole (OX), was examined for its ability to enhance brain delivery of OX compared to administration of OX in solution (OX-S). Magnetic chitosan microspheres containing OX (MCM-OX) and OX-S were administered intraarterially to male Fischer 344 rats at OX doses of 0.1 mg/kg and 0.5 mg/kg with a magnetic field of 6000 G applied to the brain for 30 min. Animals were sacrificed at 30 min and 120 min after MCM-OX and OX-S treatments, and multiple tissues were collected and analyzed for OX by HPLC. A statistical analysis of the effects of treatment, OX dose, and time on total OX in each sampled tissue was made. MCM-OX significantly increased OX brain concentrations compared to those achieved with OX-S treatments, concentrations after MCM-OX being a minimum of 100-fold greater. Within the MCM-OX treatment groups, ipsilateral OX concentrations were much greater, indicating target organ selectivity. A most interesting finding was that OX brain concentrations were similar at 120 min and 30 min after MCM-OX treatment. Thus, even in the absence of the magnetic field, MCM-OX were retained in the brain, possibly through cationic-anionic interactions with the blood-brain barrier.

Receptor-Mediated Magnetic Carriers: Basis for Targeting

A new magnetic microsphere carrier has been formulated that may localize drugs by both biochemical and physical means. The microspheres, prepared from the polysaccharide chitosan, are designed to bind to anionic glycosaminoglycan receptors on the surface of capillary endothelial cells. The microspheres were formulated to have a controlled cationic character and had a mean diameter of 0.70 µm and a magnetite content of 16% (w/w). Formation of complexes between chitosan and heparin and between the microspheres and heparin has been demonstrated. Heparin served as a model glycosaminoglycan. The chitosan:heparin complex ratio was found to be 1:1 based on charge and was formed between ammonium ions on the chitosan and SO3− groups on heparin. Neutralization of the charge on the microspheres prevented their complexation with heparin. The rationale for the use of the delivery system and its potential limitation are discussed.

Physiological pharmacokinetic modeling of inhaled trichloroethylene in rats

The pharmacokinetics of trichloroethylene (TCE) was characterized during and following inhalation exposures of male Sprague-Dawley rats. The blood and exhaled breath TCE time-course data were used to formulate and assess the accuracy of predictions of a physiologically based pharmacokinetic (PB-PK) model for TCE inhalation. Fifty or 500 ppm of TCE was inhaled by unanesthetized rats of 325-375 g for 2 hr through a miniaturized one-way breathing valve. Repetitive samples of the inhaled and exhaled breath streams, as well as arterial blood, were collected concurrently during and for 3 hr following the exposures and analyzed for TCE by headspace gas chromatography. Respiratory rates and volumes were continuously monitored and used in conjunction with the pharmacokinetic data to delineate uptake and elimination profiles. Levels of TCE in the exhaled breath attained near steady-state soon after the beginning of exposures, and were then directly proportional to the inhaled concentration. Exhaled breath levels of TCE in rats were similar in magnitude to values previously published for TCE inhalation exposures of humans. Levels of TCE in the blood of the 50 ppm-exposed animals also rapidly approached near steady-state, but blood levels in the 500 ppm-exposed animals rose progressively, reaching concentrations 25- to 30-fold higher than in the 50 ppm group during the second hour of exposure. The 10-fold increase in inhaled concentration resulted in an 8.7-fold increase in cumulative uptake, or total absorbed dose. These findings of nonlinearity indicate that metabolic saturation ensued during the 500 ppm exposure. The PB-PK model was characterized as blood flow-limited with TCE eliminated unchanged in the exhaled breath and by saturable liver metabolism. The uptake and elimination profiles were accurately simulated by the PB-PK model for both the 50 and 500 ppm TCE exposure levels. Such a model may be quite useful in risk assessments in predicting internal (i.e., systemically absorbed) doses of TCE and other volatile organics under a variety of exposure scenarios.

The uptake and elimination of 1,1,1-trichloroethane during and following inhalation exposures in rats☆☆☆

The pharmacokinetics of 1,1,1-trichloroethane (TRI) was studied in male Sprague-Dawley rats in order to characterize and quantify TRI uptake and elimination oby direct measurements of the inhaled and exhaled compound. Fifty or 500 ppm TRI was inhaled for 2 hr through a one-way breathing valve by unanesthetized rats of 325-375 g. Repetitive samples of the separate inhaled and exhaled breath streams, as well as arterial blood, were collected concurrently both during and following TRI inhalation and analyzed for TRI by gas chromatography. Respiratory rates and volumes were continuously monitored during and following exposure and were used in conjunction with the pharmacokinetic data to characterize profiles of uptake and elimination. TRI was very rapidly absorbed from the lung, in that substantial levels were present in arterial blood at the first sampling time (i.e., 2 min). Blood and exhaled breath concentrations of TRI increased rapidly after the initiation of exposure, approaching but not reaching steady state during the 2-hr exposures. The blood and exhaled breath concentrations were directly proportional to the exposure concentration during the exposures. Percentage uptake of TRI decreased 30-35% during the first hour of inhalation, diminishing to approximately 45-50% by the end of the exposure. Total cumulative uptake in the 50 and 500 ppm groups over the 2-hr inhalation exposures was determined to be 6 and 48 mg/kg body wt, respectively. By the end of the exposure period, 2.1 and 20.8 mg, respectively, of inhaled TRI was eliminated from rats inhaling 50 and 500 ppm TRI. A physiological pharmacokinetic model for TRI inhalation was utilized to predict blood and exhaled breath concentrations for comparison with observed experimental values. Overall, values predicted by the physiological pharmacokinetic model for TRI levels in the blood and exhaled breath were in close agreement with measured values both during and following TRI inhalation.

High-performance liquid chromatographic determination of the calcium channel blocker nimodipine in monkey plasma

A new high-performance liquid chromatographic (HPLC) assay was developed for the determination of nimodipine in monkey plasma. An ethyl acetate extraction procedure was employed with a reversed-phase HPLC separation for the analysis. Absolute recovery of nimodipine from plasma was over 95% with a lower limit of quantitation of 10 ng/ml. This method was applied to a preliminary pharmacokinetic study in which 0.25 mg/kg nimodipine was administered intravenously to three monkeys. Protein binding and stability of nimodipine in monkey plasma were also examined. The pharmacokinetic parameters of nimodipine in monkeys were similar to those obtained in humans and indicate that monkeys are an appropriate animal model for further pharmacokinetic investigations.

Hybrid pharmacokinetic models to describe anti-HIV nucleoside brain disposition following parent and prodrug administration in mice.

Brain delivery of active anti-HIV compounds is important for successful treatment of the AIDS patient. As an initial step in predicting human brain drug concentrations, hybrid pharmacokinetic models were developed to characterize the disposition of anti-HIV nucleosides following parent and prodrug administrations in mice. Mouse data were obtained following intravenous administration of 3′-azido-2′,3′-dideoxyuridine (AZddU or AZDU), 3′-azido-3′-deoxythymidine (AZT), and their dihydropyridine prodrugs (AZddU-DHP and AZT-DHP). Exponential equations were fitted to the serum concentration–time data for each species, including the pyridinium ion moieties, and subsequently used in differential mass balance equations describing the brain dynamics of each compound. Model parameters for the mass balance equations were estimated by various techniques, including the utilization of in vitro data. In general, model-predicted brain concentrations agreed with the observed data. Similar data in larger animals will permit scale-up of the current model to predict human brain drug concentrations.

Targeting anticancer drugs to the brain. II: Physiological pharmacokinetic model of oxantrazole following intraarterial administration to rat glioma-2 (RG-2) bearing rats

The disposition of the anticancer drug oxantrazole (OX) was characterized in rats bearing the rat glioma-2 (RG-2) brain tumor. Following intraarterial administration of 3 mg/kg of OX, serial sacrifices were completed from 5 min to 5 hr after administration. Blood and tissue samples collected at the time of sacrifice were processed and measured for OX concentrations by HPLC. The kidney had the greatest affinity for OX with the Cmaxbeing 40.6 μg/mlat 15 min after administration. OX concentrations in brain tumor were higher than in normal right and left brain hemispheres, and consistent with enhanced drug blood-tumor barrier (BTB) permeability seen in experimental models for brain tumors. Observed heart, liver, lung, and spleen OX concentrations were similar, ranging from approximately 2 μg/mlto 20 μg/ml. A unique technique was used to develop a global physiological pharmacokinetic model for OX. A hybrid or forcing function method was used to estimate individual tissue compartment biochemical parameters (i.e., partition and mass transfer coefficients). A log likelihood optimization scheme was used to determine the best model structure and parameter sets for each tissue. Most tissues required a 3-subcompartment structure to adequately describe the observed data. The global model was then reconstructed with an arterial blood and rest of body compartments that provided predicted OX concentrations in agreement with the data. The model development strategy provides a systematic approach to physiological pharmacokinetic model development.

Physiological pharmacokinetie model of adriamycin delivered via magnetic albumin microspheres in the rat

The influence of magnetic albumin microspheres on the disposition of adriamycin was evaluated. Adriamycin concentrations were monitored in multiple rat tissues for 48 hr after its intra-arterial administration (2 mg/kg) as a solution and associated with magnetic albumin microspheres. The magnetic dosage form was targeted to a predefined tail segment with a magnetic field strength of 8000 G applied for 30 min after dosing. A physiological pharmacokinetic model was used to describe the disposition of adriamycin following its administration from either dosage fcrm. The model developed for the data resulting from administration of adriamycin as a solution served as a foundation for the model developed for adriamycin resulting from the administration of adriamycin associated with the magnetic dosage form. The model for adriamycin following administration of the magnetic microspheres required additional relationships to describe the transport of adriamycin associated with the microspheres. For both models, the predicted adriamycin concentrations were in adequate agreement with the observed values. The present investigation demonstrates the use of a physiological pharmacokinetic modeling method to represent drug kinetics following its administration via a targeted drug delivery system.