Development and characterisation of soluble polymeric particles for pulmonary peptide delivery.

Pulmonary administration of protein and peptide drugs using inhaled dry powder particles is an interesting alternative to parenteral delivery. The stabilisation of these molecules is essential to the maintenance of biological activity in such inhalation formulations. Here salmon calcitonin (sCT) was co-spray dried with linear or branched PEG (L-PEG and B-PEG) and PVP in order to formulate aerosolisable particles of the bioactive peptide. Co-spray drying L-PEG and PVP resulted in porous particles, with minimal D(50) (median volume diameter) and MMAD (mass median aerodynamic diameter) values obtained for a PEG/PVP w/w ratio of 1. For particles based on both L-PEG and B-PEG, an increase in acetone, a poor solvent for the PVP, up to 70wt% of the spray dried solution led to a decrease in D(50) and MMAD. Crystallinity of PEG in the particles ranged between 90 and 97% when the PVP content varied between 15 and 70wt%, indicating a low degree of interaction between PVP and PEG. Additionally, dynamic vapour sorption analysis showed that an increase in PVP content increased the particle surface hygroscopicity. Hence, particle properties were adjusted by altering the water/acetone and PEG/PVP ratio in the spray dried solutions. PVP present at the particles surface protects them from melting during the spray drying process but also increases their hygroscopicity, adversely affecting their aerodynamic properties. Targeting a 5wt% of sCT loading resulted in a loading efficiency of 77.9 and 83.6% with L-PEG and B-PEG-based particles, respectively. Loading of sCT in L-PEG or B-PEG-based particles modified particle roughness and D(50), leading to an increase in MMAD of the L-PEG-based particles. However, particles were still considered to be suitable for aerosolisation as their FPFs (fine particle fractions) were higher than 30%. These particles formulated with PVP and PEG allowed sCT biological activity to be maintained when evaluated by measuring cAMP production by T47D cells.

Spray drying of budesonide, formoterol fumarate and their composites--I. Physicochemical characterisation.

The objective of this work was to examine the physicochemical properties of spray dried budesonide, formoterol fumarate and their mixtures at two different weight ratios: 100:6 and 400:6 of budesonide and formoterol fumarate, respectively. A comparison of the thermal properties, crystalline/amorphous nature and particle size of the starting micronised as well as processed materials was carried out. The micronised drugs on their own and the physical mixtures were crystalline in contrast to the spray dried counterparts which were shown to be amorphous. The glass transition temperatures (T(g)s) of the processed actives were determined and appeared at 89.5 and 88 degrees C for budesonide and formoterol fumarate, respectively. As for the spray dried composites, an indication of miscibility and/or interactions between the components was indicated by differential scanning calorimetry and infrared analysis. The spray drying in all cases resulted in smooth, spherical microparticles of sizes suitable for inhalation.

Spray drying of budesonide, formoterol fumarate and their composites-II. Statistical factorial design and in vitro deposition properties.

The aim of this study was to investigate the effect of changing spray drying parameters on the production of a budesonide/formoterol fumarate 100:6 (w/w) composite. The systems were spray dried as solutions from 95% ethanol/5% water (v/v) using a Büchi 191-Mini Spray Dryer. A 2(5-1) factorial design study was undertaken to assess the consequence of altering spray drying processing variables on particle characteristics. The processing parameters that were studied were inlet temperature, spray drier airflow rate, pump rate, aspirator setting and feed concentration. Each batch of the resulting powder was characterised in terms of thermal and micromeritic properties as well as an in vitro deposition by twin impinger analysis. Overall, the parameter that had the greatest influence on each response investigated was production yield - airflow (higher airflow giving greater yields), median particle size - airflow (higher airflow giving smaller particle sizes) and Carr's compressibility index - feed concentration (lower feed concentration giving smaller Carr's indices). A six- to seven-fold difference in respirable fraction can be observed by changing the spray drying process parameters. The co-spray dried composite system which displayed best in vitro deposition characteristics, showed a 2.6-fold increase in respirable fraction in the twin impinger experiments and better dose uniformity compared with the physical mix of micronised powders.

Characterisation of excipient-free nanoporous microparticles (NPMPs) of bendroflumethiazide.

The purpose of this study was to prepare excipient-free porous microparticles of bendroflumethiazide by spray drying and to characterise the physicochemical properties of the particles produced. Solutions of bendroflumethiazide in ethanol/water, ethanol/water/ammonium carbonate or methanol/water/ammonium carbonate were spray dried using a laboratory spray dryer. Spray dried products were characterised by scanning electron microscopy, X-ray powder diffraction, differential scanning calorimetry, FTIR, laser diffraction particle sizing and density measurement. Nanoporous microparticles (NPMPs) were prepared from the alcoholic solutions containing ammonium carbonate. NPMPs were amorphous in nature, had median particles sizes less than 3 microm and densities that were significantly reduced compared to non-porous spray dried bendroflumethiazide powder. The novel process may be used to produce excipient-free amorphous microparticles with desirable physical properties such as amorphous solid state, porosity and low bulk density. This new engineering technology has applications in the design of other therapeutic agents such as those used in pulmonary delivery.

Release kinetics of benzoic acid and its sodium salt from a series of poly(N-isopropylacrylamide) matrices with various percentage crosslinking.

Swelling and concomitant drug-release kinetics from a series of poly(N-isopropylacrylamide) (PNIPA) matrices were examined. Scanning electron microscopy indicated a decrease in polymer pore/mesh size above the lower critical solution temperature (LCST) with increasing percentage crosslinker. The release of sodium benzoate (NaB) or benzoic acid (BA) were investigated above and below the LCST of the gels and compared to the drug-loaded gel-swelling rates. The release rate of NaB increased with increasing percentage crosslinker above the LCST in contrast to a decrease in release rate with increasing crosslinker seen previously with non-thermoresponsive hydrogel systems. As the percentage crosslinker increased, there was therefore a decrease in the ability to thermally control the release of this small model drug. In contrast to the crosslinker-dependent pattern apparent with NaB, drug-PNIPA hydrophobic binding controlled the swelling rate of BA-loaded hydrogels. As a result, all the BA-loaded systems showed similar diffusion controlled swelling and release patterns, effectively independent of the inherent-swelling rates of the hydrogels. The crosslinking content of the hydrogel and the physicochemical nature of the loaded drug were therefore shown to be important in thermal control of drug release from PNIPA hydrogels.

Combined effects of iontophoretic and chemical enhancement on drug delivery. II. Transport across human and murine skin.

This paper reports measurements of the release characteristics of the model drug salbutamol from a liquid crystalline vehicle across both human and hairless murine skin in vitro. The use of oleic acid and iontophoresis as penetration enhancement techniques, used separately and simultaneously, was also investigated. Over a period of 12h, salbutamol base did not diffuse from the vehicle across excised human skin while, in contrast, over a period of 2h, the drug passively transported across hairless murine skin. The diffusion co-efficient for the drug in this tissue was estimated to be 4.54+/-0.60x10(-9)cm(2)s(-1) with a permeability co-efficient of 7.03+/-0.83x10(-7)cms(-1). A current of density of 0.39mAcm(-2) facilitated a significant transport of salbutamol from the liquid crystalline vehicle across excised human skin but with a small (<0.1) transport number. The quantity of salbutamol transported across excised hairless murine skin under the same conditions was significantly greater with a transport number of 0.68. The alteration of the permeability of the tissue was less than that of the human skin and a full recovery of the pre-iontophoretic permeability of murine skin was consistently observed. The incorporation of either oleic or lauric acid into the monoglyceride component of the vehicle at a concentration of 0.1M had a marked effect on the transport of salbutamol across both human and murine skin. The initial passive permeation of the drug across the skin was not affected but the rate of drug delivery during iontophoresis was typically observed to increase by a factor greater than two. The post-iontophoretic transport of salbutamol across either tissue was also substantially enhanced in the presence of the fatty acid. The analogous use of stearic acid did not significantly influence the iontophoretic or the post-iontophoretic transport of salbutamol across excised human skin. The investigation also revealed a synergistic combination of the fatty acid and anodal iontophoresis to enhance the in vitro transport of other drug substances, including nicotine and diltiazem hydrochloride across murine skin. Oleic acid increased both the iontophoretic and post-iontophoretic transport of nicotine, so that the enhancement of drug delivery was greater than that caused by the current alone. The investigation also indicated that the barrier properties of the skin recover following the constant current iontophoresis in the presence of oleic or lauric acids.

Nanoparticles: pharmacological and toxicological significance.

Nanoparticles are tiny materials (<1000 nm in size) that have specific physicochemical properties different to bulk materials of the same composition and such properties make them very attractive for commercial and medical development. However, nanoparticles can act on living cells at the nanolevel resulting not only in biologically desirable, but also in undesirable effects. In contrast to many efforts aimed at exploiting desirable properties of nanoparticles for medicine, there are limited attempts to evaluate potentially undesirable effects of these particles when administered intentionally for medical purposes. Therefore, there is a pressing need for careful consideration of benefits and side effects of the use of nanoparticles in medicine. This review article aims at providing a balanced update of these exciting pharmacological and potentially toxicological developments. The classes of nanoparticles, the current status of nanoparticle use in pharmacology and therapeutics, the demonstrated and potential toxicity of nanoparticles will be discussed.

Preparation and release of model drugs from thermally sensitive poly(N-isopropylacrylamide) based macrospheres.

Emulsion polymerization was employed to prepare poly(N-isopropylacrylamide) hydrogel spheres, which exhibited an LCST of 32 degrees C. The hydrogels were loaded with model drugs (benzoic acid (BA), sodium benzoate and diltiazem HCl (DHCl)) and release investigated at 25 degrees C and 37 degrees C. The temperature at which gel formation occurred was vital for successful hydrogel preparation, macrosphere formation not occurring when the temperature was close to the LCST. Sphere size increased on decreasing the stirring rate and on slowing the rate of addition of the aqueous phase. Pulsatile delivery was investigated using BA and DHCl. For both compounds a pulse was observed with a change in temperature. Pulsed release of the smaller model drug of lowest solubility, BA, was more successful. Drug release from hydrogel spheres was, therefore, found to be dependent on the physicochemical properties of the drugs, with pulsatile release of low molecular weight compounds, by temperature cycling, difficult to control.

Drug-polymer interactions and their effect on thermoresponsive poly(N-isopropylacrylamide) drug delivery systems.

Potential interactions between model drugs (benzoates, diltiazem, cyanocobalamin, dextrans) and a thermoresponsive poly(N-isopropylacrylamide) (PNIPA) hydrogel and corresponding linear polymer were investigated. The influence of the drugs on the equilibrium swelling level of the hydrogel was examined and drug-hydrogel binding isotherms were established where appropriate. Differential scanning calorimetry (DSC) was used to investigate the influence of the drugs on the lower critical solution temperature (LCST) of the linear polymer solution. Phase solubility studies were preformed to investigate binding. Drug-polymer co-precipitated blends were also prepared and analysed by X-ray diffraction (XRD), thermal analysis and Fourier transform infrared (FT-IR) spectroscopy. Hydrophobic binding was apparent between PNIPA and the aromatic ring/ester side chain of the unionised benzoate. The effect of this binding on hydrogel swelling was clarified in terms of the influence of the binding on the LCST of the system. The drug release rates of the benzoates from the hydrogel were shown to be dependent on drug binding properties. Ionisation of the benzoate prevented such hydrophobic binding, with a weaker salting out effect apparent with sodium benzoate. Significant interactions between diltiazem, cyanocobalamin (Vitamin B12) or the dextrans and PNIPA were not apparent. High concentrations of the hydrophilic drugs did, however, interfere with the magnitude of hydrogel equilibrium swelling. Hydrophobic binding, the salting out effect and the influence of the drugs on hydrogel swelling under non-sink conditions were therefore shown to be important effects which depended on the chemical nature of the drug present.

Hydrodynamic simulation (computational fluid dynamics) of asymmetrically positioned tablets in the paddle dissolution apparatus: impact on dissolution rate and variability.

The aim of this work was to investigate the dissolution rate from both the curved and planar surfaces of cylindrical compacts of benzoic acid, which were placed centrally and non-centrally at the base of the vessel of the paddle dissolution apparatus. The effect of fixing the compacts to a particular position on the variability of dissolution results was also examined. In addition, computational fluid dynamics (CFD) was used to simulate fluid flow around compacts in the different positions in the vessel, and the relationship between the local hydrodynamics in the region of the compacts and the dissolution rate determined. The dissolution rate was found to increase from the centre position to the off-centre positions for each surface examined. There was a corresponding increase in maximum fluid velocities calculated from the CFD fluid flow simulations at a fixed distance from the compact. There was less variability in dissolution from compacts fixed to any of the positions compared with those that were not fixed. Fluid flow around compacts in different positions could be successfully modelled, and hydrodynamic variability examined, using CFD. The effect of asymmetric fluid flow was evident visually from the change in shape of the eroded compacts.

Effect of drug physicochemical properties on swelling/deswelling kinetics and pulsatile drug release from thermoresponsive poly(N-isopropylacrylamide) hydrogels.

The effect of drug physicochemical properties on swelling/deswelling kinetics and pulsatile drug release from a thermoresponsive hydrogel was examined. Hydrogels were loaded with drug and thermally triggered swelling/deswelling and release experiments were performed. Two series of drugs of contrasting hydrophilicity and varying physicochemical properties were examined. Benzoic acid (BA), its methyl and propyl esters, and diltiazem base were used as model hydrophobic drugs. Sodium benzoate (NaB), diltiazem HCl (DHCl), vitamin B12 (VB12) and various dextrans (MW 4300, 10,200, 42,000, 68,800) were used as model hydrophilic agents of increasing size. The hydrogel swelling rate was slowed by the presence of the hydrophobic drugs and this decreased rate was solubility dependant for the benzoates. The hydrophilic series increased the rate of swelling compared to the unloaded system. In all cases, the magnitude and rate of hydrogel contraction were proportional to the extent of swelling prior to temperature switch. Drug release was by diffusion below the lower critical solution temperature (LCST), while a solubility-dependent drug pulse release on temperature switch was observed for the hydrophobic series. Effectiveness of thermal control of hydrophobic drug release increased with increasing solubility. The hydrophilic series produced a molecular size-dependent drug pulse on temperature switch above the LCST. Pulsatile on-off drug release was shown with DHCl, VB12 and the various dextrans. Drug solubility, size and chemical nature were shown to be of particular importance in the control of hydrogel swelling and drug release from thermosensitive hydrogels.

Iontophoretic and chemical enhancement of drug delivery. Part I: across artificial membranes.

This paper reports on measurements of the release characteristics of the model drug salbutamol base from a liquid crystalline vehicle across a non-rate limiting synthetic membrane. The measured passive release rates were compared with analogous behaviour: (i) when a penetration enhancer such as oleic acid was incorporated into the vehicle; (ii) when the release was iontophoretically assisted; and (iii) when the penetration enhancer and iontophoretic assistance were used simultaneously. The effects of using isotonic phosphate buffer solution as the aqueous domain of the vehicle and in the receptor were also separately assessed. The passive release from the standard system was consistent with matrix diffusion control. The addition of oleic acid indicated association of the drug with the fatty acid so that its release into an aqueous medium was significantly retarded. With buffer ions present in the vehicle the release rate increased consistent with reduced association, and when phosphate buffer was used as a receptor medium the release rate exceeded that of the standard vehicle due to an ion exchange process. The delivery of salbutamol from the fatty acid containing systems was substantially enhanced by iontophoresis and the rates were shown to be approximately proportional to the assisting currents. The data clearly indicate the iontophoretic process to be significantly less efficient in the presence of buffer ions but with the iontophoretic delivery rates being enhanced by the presence of a fatty acid.

Sensitivity of dissolution rate to location in the paddle dissolution apparatus.

The aim of the present study was to determine the apparent diffusion boundary layer and dissolution rate constant for various surfaces of compacts and at various locations in the USP paddle dissolution apparatus. Benzoic acid compacts were coated with paraffin wax leaving only the surface under investigation free for dissolution. The dissolution rates for various surfaces at varying locations in the paddle dissolution vessel were determined from the slope of the dissolution profile (amount dissolved (mg) versus time (min)). The apparent diffusion boundary layer and dissolution rate constant were calculated and were found to vary depending on the surface of the compact from which dissolution took place and also on the location and size of the compact. It may be concluded that, in developing models to describe the dissolution from solid dosage forms, it is not accurate to assume constant hydrodynamics and mass transfer rates at all surfaces of the system, or in different locations within the test device. A more exact description of the hydrodynamics would be necessary in order to precisely model drug dissolution in the paddle dissolution apparatus.

The problem of bias in training data in regression problems in medical decision support.

This paper describes a bias problem encountered in a machine learning approach to outcome prediction in anticoagulant drug therapy. The outcome to be predicted is a measure of the clotting time for the patient; this measure is continuous and so the prediction task is a regression problem. Artificial neural networks (ANNs) are a powerful mechanism for learning to predict such outcomes from training data. However, experiments have shown that an ANN is biased towards values more commonly occurring in the training data and is thus, less likely to be correct in predicting extreme values. This issue of bias in training data in regression problems is similar to the associated problem with minority classes in classification. However, this bias issue in classification is well documented and is an on-going area of research. In this paper, we consider stratified sampling and boosting as solutions to this bias problem and evaluate them on this outcome prediction problem and on two other datasets. Both approaches produce some improvements with boosting showing the most promise.

Preparation and characterisation of a range of diclofenac salts.

Physicochemical properties of diclofenac salts prepared using eight different counterions and including five novel salts, obtained with the bases 2-amino-2-methyl-1,3-propanediol, 2-amino-2-methylpropanol, tert-butylamine, benzylamine and deanol, were compared. Four of the bases used to prepare these salts were related in their chemical structure, differing only in the number of hydroxy groups. Characterisation techniques included X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis, thermomicroscopy, Karl Fischer titration, FT-IR spectroscopy and elemental analysis. In the case of salts prepared from 2-amino-2-methylpropanol and benzylamine, two polymorphic forms of each salt were identified. For the 2-amino-2-methyl-1,3-propanediol salt, a pseudopolymorphic form was identified. The aqueous solubilities of the salts studied ranged from 3.95 mM (tris(hydroxymethyl)aminomethane salt) to 446 mM (deanol salt), corresponding to a 113-fold difference in solubility. The solubility of diclofenac deanol was higher than the solubilities for diclofenac salts reported earlier. Correlation was found between the inverse of the salt melting point and the logarithm of salt solubility. A log-log relationship was observed between salt solubility and hydrogen ion concentration in the salt solution. Relationships between the properties of the salt-forming agents and those of the resulting diclofenac salts were explored. Reasonable correlation was found between the free base melting point and the salt melting point.

Factors influencing drug release from stearic acid based compacts.

Fatty acids are potentially suitable carriers for use in the design of drug delivery systems, being biocompatible, biodegradable inexpensive and of low toxicity. The release of the model compound benzoic acid from fatty acid compacts of stearic acid was evaluated using the USP Apparatus 2 dissolution assembly in phosphate buffer pH 7.4. Matrix controlled drug release was expected. Release profiles were approximated by square root of time kinetics. Release rate was independent of stirring speed in the rpm range 50-150, however, at 200 rpm a significant increase in release rate was observed particularly at later times, the amount released versus square root of time plots becoming non-linear. Release was independent of compression pressure in the range 1-7 tons. The particle size of the benzoic acid and stearic acid used had a significant influence on release. The use of particles in the range 250-500 microm gave release rate constants (k, g/cm(2) per min(0.5)) approximately 1.5 greater than those of smaller particle size (63-125 microm). The formation factor (F) tended to increase exponentially with drug loading, the increase being steeper for compacts prepared from the larger particle sizes. At 80% drug loading for large sized systems the matrix appeared to offer little resistance to drug release and F approached one.

Comparison of the physicochemical properties of the N-(2-hydroxyethyl) pyrrolidine, diethylamine and sodium salt forms of diclofenac.

Non steroidal anti-inflammatory agents (NSAIDs) such as diclofenac have very low aqueous solubilities and consequently salt formation may be used to enhance solubility and dissolution rate. In this study, we examined the physicochemical properties of three diclofenac salts, diclofenac sodium (DNa), diclofenac N-(2-hydroxyethyl)pyrrolidine (DHEP) and diclofenac diethylamine (DDEA), and their different solid state forms to determine the influence of salt form on solubility, dissolution rate and membrane transport. The equilibrium solubility of DDEA at 25 degrees C was determined as 33 mM, lower than the solubilities of DHEP (273 mM) and DNa (66 mM) previously reported (Ledwidge and Corrigan, 1998). In addition to the dihydrate form of DHEP previously characterised, monohydrate forms of DHEP and DDEA were identified. Intrinsic dissolution rate studies were used to determine the solubility ratios of the hydrated and anhydrous forms. The monohydrate form of DHEP was found to be 1.8 times less soluble than the anhydrate, whereas DDEA anhydrate was approximately 1.7 times as soluble as the monohydrate form. On investigation of the pH-solubility profile (25 degrees C) of DDEA, appreciable supersaturation (76 mM) relative to the theoretical profile, was detected at the pH(max). This contrasts with values of >800 and 67 mM for DHEP and DNa, respectively. The transport of salt solutions through a porous membrane (Visking) was investigated. A linear relationship between concentration (mM) and rate of transport (mmol/h) was established for DNa and DHEP solutions. The mass transfer coefficient determined for DHEP was lower than that for the other two salts. Nevertheless, the maximum transport rate obtained for DHEP is almost six times higher than that obtained for DDEA.

Characterization of drug release from diltiazem-loaded polylactide microspheres prepared using sodium caseinate and whey protein as emulsifying agents.

The influence of milk protein emulsifying agents on the characteristics, particularly drug release, of polylactide microspheres was investigated. Diltiazem loaded polylactide (PL) microspheres were successfully prepared using the dairy proteins, sodium casinate (SC) and whey protein isolate (WPI) as the emulsifying agents. Microspheres were characerized in terms of microsphere yield, electron microscopy, particle size, drug loading, DSC and XRD analysis and drug release. The yields of microspheres obtained were 53-63% and were independent of the emulsifying agent used. SEM revealed that, regardless of the emulsifying agent employed, the microspheres were of good sphericity, but the surface appearance of the microspheres was not the same in all cases. The milk proteins resulted in microspheres approximately half the size of those obtained with methylcellulose (MC). Significant differences in drug loading were observed between the three emulgents, the MC systems giving the highest values. Release profiles were sigmoidal in shape and were well fitted to the equation ln (x/1 - x) = k x t - k x tmax, reflecting degradation controlled drug release. The parameter k increased with drug loading, while tmax decreased. The relationships between the release parameters [P(k and tmax)] and loading (L) could be quantified by equations of the form P = a x L(N), N being negative in the case of tmax. Apart from the effect on loading efficiency, neither SC nor WPI appeared to significantly alter drug release. The quantitative relationships observed in this study may have more general application in quantifying drug release from drug-polymer composites at low loadings where polymer degradation controls drug release.

Mechanistic aspects of the release of levamisole hydrochloride from biodegradable polymers.

The release of levamisole hydrochloride from poly-DL-lactide-co-glycolide compacts prepared at 5, 10 and 20% drug loading using two different particle size fractions of drug (90-125 and 125-250 microm) was investigated. Release profiles were significantly different from those previously reported for compacts prepared using the base form of the drug. Release was found to occur in a biphasic manner, with an initial fast release phase followed by a slower polymer degradation controlled release phase. The drug release profiles were successfully described by a model combining contributions from a first-order initial release phase and a polymer degradation controlled drug release phase. The fraction of drug released in the initial burst phase (F(B)) was attributed to the dissolution of drug domains situated at the surface of the polymer-drug compact and this fraction tended to increase with increasing drug particle size, as expected from the model. The increase in F(B) with increased loading was attributed to the clumping of dispersed drug particles which effectively increased the proportion of drug linked to the compact surface.

Preparation and evaluation of microspheres prepared from whey protein isolate.

Whey protein (WPI) microspheres were successfully produced containing hydrochlorothiazide, eosin, patent blue violet and sodium salicylate using a w/o emulsification method with glutaraldehyde cross linking. The release of these compounds from WPI microspheres occurred rapidly, with at least 70% of the incorporated material released for all systems within the first 20 min. Release of microsphere payload was essentially complete within 1 h. The degree of glutaraldehyde cross-linking was found to have no effect on the release profile for durations of cross-linking up to 24 h. Of a range of release equations examined, the experimental release data was best described by a biexponential equation and this agrees with the work of Tomlinson et al. (1984) for the release of drugs from albumin microspheres. Swelling of the microsphere systems was examined as this may contribute to the rapid release of drug from these systems.