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Novel biodegradable and mucoadhesive PLGA/chitosan microparticles with the potential for use as a controlled release gastroretentive system were manufactured using supercritical CO(2) (scCO(2)) by the Particle Gas Saturated System (PGSS) technique (also called CriticalMix(TM)).Microparticles were produced from PLGA with the addition of mPEG and chitosan in the absence of organic solvents, surfactants and crosslinkers using the PGSS technique. Microparticle formulations were morphologically characterized by scanning electron microscope; particle size distribution was measured using laser diffraction. Microparticle surface was analyzed using X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) to evaluate the presence of chitosan on the surface. Mucoadhesiveness of the microparticles was evaluated in vitro using a mucin assay employing two different kinds of mucin (Mucin type III and I-S) with different degrees of sialic acid contents, 0.5-1.5% and 9-17%, respectively.The two analytical surface techniques (XPS and ToF-SIMS) demonstrated the presence of the chitosan on the surface of the particles (100 μm), dependent on the polymer composition of the microparticles. The interaction between the mucin solutions and the PLGA/chitosan microparticles increased significantly with an increasing concentration of mucin and chitosan.The strong interaction of mucin with the chitosan present on the surface of the particles suggests a potential use of the mucoadhesive carriers for gastroretentive and oral controlled drug release.

The surfaces of polystyrene microspheres (60 nm in diameter) and colloidal gold particles (17 nm in diameter) were coated with a polyoxyethylene (POE)/polyoxypropylene (POP) block co-polymer; poloxamine-908. The polymer adsorb strongly to the microspheres via its relatively hydrophobic POP segments. This leaves the POE chains in a mobile state as they extend outward from the surface and thereby provide stability to the particle suspension by suppressing aggregation. The blood clearance and biodistribution of uncoated vs. poloxamine-908-coated 125I-labelled polystyrene microspheres were compared 1 h after intravenous administration into rats. Poloxamine coating dramatically reduced liver accumulation of microspheres and kept them within the systemic circulation. These observations were further confirmed by electron microscopy, demonstrating that Kupffer cells were loaded with uncoated latex but had ingested few if any of the poloxamine-908-coated particles. The interaction of uncoated and poloxamine-coated gold particles with freshly isolated rat liver sinusoidal cells was examined by electron microscopy. The accumulation in Kupffer cells of gold particles after opsonization with autologous plasma was in accordance with previous observations where the dominant opsonizing activity had been identified as fibronectin. In contrast, coating of gold particles with poloxamine-908 prior to plasma opsonization prevented the adsorption of fibronectin onto their surface. Simultaneously, Kupffer cells failed to recognize poloxamine-908-coated gold particles before and after opsonization. Unlike Kupffer cells, liver endothelial cells endocytosed poloxamine-908-coated gold particles prior to opsonization but failed to recognize them after the opsonization process. This was taken as an indication of the presence of dysopsonic activity in plasma. This dysopsonic activity was studied using polystyrene latex microspheres, where the uptake of such particles by phagocytes is known to be independent of opsonization. The coating of 125I-labelled polystyrene microspheres with poloxamine-908 dramatically reduced their interaction with liver sinusoidal cells. This interaction was further reduced in the presence of either autologous plasma or serum. A heat-stable (60 degrees C for 15 min) serum component of molecular mass > 100 kDa was found to mediate this suppressive effect. Thus, we demonstrate that organ-specific receptors, opsonin activities and plasma dysopsonins regulate the in vivo clearance of particulate materials from the circulation. Poloxamine-908 coating modulates particle clearance by effectively blocking opsonization but still allowing for dysopsonization.

The biodistribution of injected colloidal carriers for targeted delivery will be highly dependent upon their size and surface properties. This paper describes investigations on the preparation of sub-200 nm poly(lactide-co-glycolide) (PLGA) microspheres by the variation of processing parameters using the solvent evaporation technique. Smaller particle sizes were found to be favoured by a two stage emulsification process, a low PLGA concentration and an increased surfactant concentration. In the latter case, it would appear that the viscosity of the continuous phase is a crucial factor. The process of particle size reduction could further be complimented by using a surfactant of lower molecular weight. Employing such procedures, microspheres as low as 90 nm in diameter of low polydispersity were prepared in a reproducible manner.

The molecular weights and gross aqueous solution conformation of two chitosans of different degrees of acetylation, 'Sea Cure +210' (11% acetylated) and KN50 (58% acetylated) were characterized by viscometry, analytical ultracentrifugation and dynamic light scattering. The hydrodynamic parameters obtained were used to determine both the molecular weights and the gross solution conformation of the two chitosans. Using the Wales-Van Holde ratio of sedimentation coefficient concentration regression coefficient (ks) to intrinsic viscosity [eta], the Sea Cure +210 chitosan, which is much less acetylated than the KN50, is highly asymmetric in conformation. This, in conjunction with the charge on the molecule, would suggest a rod-like conformation in solution. The two largest KN50 chitosans have widely differing values for the Wales-Van Holde ratio, suggesting different solution conformation. However, when the series is examined as a whole using the Mark-Houwink-Kuhn-Sakurada relationships relating molecular weight to both intrinsic viscosity and translational diffusion coefficient, then a more spheroidal structure, approximating to a random coil, is predicted.

SDS-PAGE in combination with densitometry has been used to evaluate the adsorption of plasma and serum proteins to polystyrene microspheres (PS) coated with block copolymers of the poloxamer and poloxamine series. The protein-resistant nature of coated PS was demonstrated for these systems when incubated in dilutions of plasma and serum. The total amount of protein and the type of proteins adsorbed were dependent on the plasma and serum incubation concentration used. At 0.3% serum concentrations the total amount of protein adsorbed was found to be related to the polyoxyethylene (PEO) chain length of the block copolymer, whilst at 0.3% or 50% plasma concentrations a relationship was shown between the polyoxypropylene (PPO) chain and the plasma protein adsorption for the range of block copolymers studied. Immunoblotting studies revealed the adsorption of immunoglobulin G, complement C3, transferrin and fibronectin to all microspheres previously incubated in 50% serum and plasma, whilst fibrinogen was also adsorbed after incubation in 50% plasma; with similar quantities of each protein adsorbed to PS and block copolymer-coated PS.

Human serum albumin (HSA) nanospheres of about 100 nm diameter were prepared using a pH-coacervation method whereby acetone was added to an HSA solution (pH 9.0). The particles obtained were cross-linked by glutaraldehyde. Increasing the pH of the HSA solution resulted in a gradual rise in the particle size of the resultant nanospheres. A higher cross-linking efficiency was obtained with increased glutaraldehyde concentration and cross-linking time. No significant differences in surface properties, as determined by zeta potential measurements, were recorded between particles prepared from HSA solutions with different pH. The nanospheres were quite stable over 4 days in both phosphate buffer saline (PBS) solution (pH 7.4) and rat serum, but degraded rapidly over 6 hours when incubated in PBS solution containing trypsin.

Present work investigates the potential of chitosan nanoparticles, formulated by the ionic gelation with tripolyphosphate (TPP), to open the cellular tight junctions and in doing so, improve the permeability of model macromolecules. A comparison is made with chitosan solution at equivalent concentrations. Initial work assessed cytotoxicity (through MTS and LDH assays) of chitosan nanoparticles and solutions on Calu-3 cells. Subsequently, a concentration of chitosan nanoparticles and solution exhibiting minimal toxicity was used to investigate the effect on TEER and macromolecular permeability across filter-cultured Calu-3 monolayer. Chitosan nanoparticles and solution were also tested for their effect on the distribution of the tight junction protein, zonnula occludens-1 (ZO-1). Chitosan nanoparticles produced a sharp and reversible decrease in TEER and increased the permeability of two FITC-dextrans (FDs), FD4 (MW 4 kDa) and FD10 (MW 10 kDa), with effects of a similar magnitude to chitosan solution. Chitosan nanoparticles produced changes in ZO-1 distribution similar to chitosan solution, indicating a tight junction effect. While there was no improvement in permeability with chitosan nanoparticles compared to solution, nanoparticles provide the potential for drug incorporation, and hence the possibility for providing controlled drug release and protection from enzymatic degradation.

Insulin was administered vaginally to sheep as an aqueous solution and as a lyophilised powder with bioadhesive starch microspheres. The effect of lysophosphatidylcholine (LPC) on the vaginal absorption of insulin from both formulations was studied. While the vaginal absorption of insulin from insulin solution was minimal, the addition of LPC resulted in a rapid rise in plasma insulin and a pronounced fall in plasma glucose levels. The absolute bioavailability of the peptide from the latter solution was 13%. The hypoglycaemic response to vaginally administered insulin was also improved using the microsphere delivery system, compared to insulin solution alone, and was further enhanced by LPC. Vaginal absorption of insulin from each formulation appeared to be influenced by the oestrous cycle and was thought to correlate with changes in vaginal histology.

A series of fractionated di-iodo-l-tyrosine-labelled dextrans (DIT dextrans), with a narrow range of number average molecular weights from 1260 to 45 500 Da, was administered intranasally and intravenously to anaesthetized rats. The nasal absorption of these compounds ranged from 0·6 to 52·7%. There was an inverse relationship between molecular size and the proportion of an intranasal dose absorbed. The study demonstrated the usefulness of DIT dextrans as molecular weight markers and confirmed the relationship between molecular size and nasal absorption for highly water soluble compounds. These results also supported the proposition that there is a continuous range of aqueous pores in the nasal mucosa.

Samples of poloxamer 407 obtained from different suppliers have been assessed for their ability to redirect model polystyrene colloids to the bone marrow in rabbits. Only poloxamer 407 obtained from one supplier was found to produce the targeting effect seen previously. The variation in the biodistribution patterns produced by the different samples of poloxamer 407 may by caused by differences in the molecular weight distribution of the polymer.

The effect of charge reversal on the uptake of fat emulsions by the reticuloendothelial (RE) system has been assessed by in vitro and in vivo methods. Emulsions stabilized by egg lecithin were charge reversed from negative to positive by added calcium ions that are adsorbed at the emulsion surface. The interaction of positively charged emulsion droplets with rat Kupffer cells was not significantly different to that found with negatively charged emulsion droplets. In all cases the measured uptake was low. Blockade of the reticuloendothelial system by infused fat emulsion was evaluated in vivo using a rabbit model. RE function was quantified by means of a probe in the form of radiolabelled albumin microspheres together with gamma scintigraphy. The kinetics of uptake of the probe into the liver/spleen regions and organ analysis showed that there were no differences between conventional and charge reversed systems. The extent of RE blockade was small in all cases.

Over the last twenty years, more attention has focused on the vagina as a favourable alternative site for the systemic delivery of therapeutic peptides and proteins. The present review aims to discuss the anatomy and physiology of the vagina and the factors which will affect drug transport across the vaginal epithelium. Emphasis is also placed on the characteristics and suitability of various animal models used in the study of vaginal absorption. The literature concerning the vaginal absorption of luteinizing hormone-releasing hormone and its analogues and insulin is reviewed in detail together with work performed using insulin in our laboratories. Due to the physicochemical characteristics of polypeptides and the potential for enzymatic degradation, vaginal absorption of these drugs is normally low. Hence, the need for absorption enhancers in the administration of polypeptides and their histological implications are discussed. Finally, the need for the development of novel delivery systems suitable for peptide administration is also stressed.

The absorption of insulin across the vaginal mucosa into the systemic circulation was studied in ovariectomized rats given subsequent estrogen treatment. Blood glucose levels were determined as an indirect measure of insulin absorption, and the effect of various enhancers on the hypoglycemic response was investigated. In the absence of any enhancer, no decrease in blood glucose levels was observed after vaginal administration of insulin. However, the coadministration of sodium taurodihydrofusidate, polyoxyethylene-9-lauryl ether, lysophosphatidylcholine, palmitoylcarnitine chloride, and lysophosphatidylglycerol significantly increased hypoglycemia, whereas citric acid had little effect. The histological changes in the vaginal epithelium after treatment with the enhancer systems were variable and often severe. While the efficacy of these compounds in promoting the vaginal absorption of insulin is encouraging, their mechanisms of action and long-term histological effects are yet to be defined.

Dextran is currently being considered as a carrier biopolymer system for drug targeting. Di-iodotyrosine (DIT), and its radioactive derivative are useful markers for identifying the in-vivo route of these substrates. The performance of these substances in vivo is closely linked to their physico-chemical properties in solution. These properties have been investigated on a series of gel permeation chromatography fractionated DIT-dextrans using a combination of sedimentation, viscometric and laser-light scattering techniques. Weight average molecular weights, Mw, from sedimentation equilibrium and light scattering range from 0·0195 to 0·054 × 106. Double logarithmic representations of Mw versus sedimentation coefficient, so20.w, and intrinsic viscosity [η] suggest that these labelled dextrans have a more compact conformation in solution than do their unlabelled analogues — which behave as random coils. The implications of this for drug delivery are indicated.

Novel sustained release formulations of hGH prepared by supercritical fluid processing of PLGA/PLA (the CriticalMix process) were produced in the form of microparticles for subcutaneous injection. The basis of the process is that PLGA/PLA polymers liquefy when exposed to supercritical CO(2), thereby allowing the hGH to be mixed efficiently into the polymers at an ambient temperature and in the absence of solvents. The CO(2) was removed from the mixture by depressurisation through a nozzle, resulting in the production of microparticles containing the hGH, which were collected in a cyclone. The best microparticle formulations showed an initial in vitro burst of around 35% and a sustained release over 14 days. When tested in the rat model, which displays a faster clearance rate of hGH than other animal models, two formulations showed prolonged release over 2-3 days with sustained plasma levels at 1-5 ng/ml whereas the soluble hGH formulation was cleared within 24h. Two selected sustained release formulations were tested in cynomolgus monkeys and compared to a single injection of soluble hGH. The burst release from the sustained release formulations was similar in magnitude to a daily dose of hGH and serum hGH levels were maintained for a seven day period. It is probable from the data that the sustained release would have continued for up to 14 days if sampling had been continued. The IGF-1 results showed there was no significant difference between the levels obtained for once daily injection of soluble hGH and the two sustained release formulations.

This review aims to evaluate the evidence for the existence of a direct nose-to-brain delivery route for nanoparticles administered to the nasal cavity and transported via the olfactory epithelium and/or via the trigeminal nerves directly to the CNS. This is relevant in the field of drug delivery as well as for new developments in nanotechnology. Experiments in animal models have shown that nano-sized drug delivery systems can enhance nose-to-brain delivery of drugs compared to equivalent drug solutions formulations. Protection of the drug from degradation and/or efflux back into the nasal cavity may partly be the reason for this effect of nanoparticles. It is uncertain, however, whether drug from the nanoparticles is being released in the nasal cavity or the nanoparticles carrying the drug are transported via the olfactory system or the trigeminal nerves into the CNS where the drug is released. Furthermore, toxicity of nanoparticulate drug delivery systems in the nasal cavity and/or in the CNS has not been extensively studied and needs to be considered carefully.

The effect of L-α-lysophosphatidylcholine (LPC) on the nasal absorption of human growth hormone (hGH) in anaesthetised rats. conscious rabbits and sheep, has been examined. LPC was shown to be an effective enhancer of nasal hGH absorption in three different animal models. In the rat. concentrations of LPC in the range 0–1.0% co-administered with hGH gave absorption values, relative to subcutaneous administration (S/C), from 2.3 to 16.3%. In the rabbit, concentrations of LPC of 0 and 0.2% co-administered with hGH gave absorption values, relative to S/C. of 1.4 and 72.8% respectively. In the sheep, concentrations of LPC in the range 0–0.59% co-administered with hGH gave absorption values, relative to S/C, from 0.2 to 16.0%. The amount of hGH absorbed was directly proportional to the concentration of LPC co-administered, and in sheep this relationship was linear, with a correlation coefficient of 0.99. The shapes of the plasma concentration vs time curves were similar in all three species.

A method is described which enables the controlled assessment of the effects of intranasal drug formulations on rat nasal epithelium. Damage to the nasal mucosa caused by the surfactant laureth-9 (1% w/v) was used as an indicator of the distribution of dose volumes (100, 50 and 20 μl) administered to one side of the nasal cavity. Only the 20 μl volume was retained exclusively in the dosed side. Using this dose volume therefore, the epithelium on the undosed side could be used as control tissue and a direct comparison made between experimental and control tissue in the same section. Errors due to inter-animal variablity and tissue sampling are thus minimised, and the total number of animals required for future studies reduced.

In accordance with our previous studies in the rabbit (Ilium L. and Davis S.S., Life Sci., 40 (1987) 1553–1560), coating of model polystyrene microspheres (70 nm in size) with poloxamer-407 was found to prevent their hepatic sequestration and to redirect a significant proportion of intravenously injected particles to the bone marrow in rats. In contrast, preincubation of a stable and ‘solid’ small unilamellar liposome preparation (DSPC/Chol/DCP in a molar ratio of 7: 7:1) with poloxamer-407 provided no evidence of poloxamer penetration or adsorption onto the vesicles, as determined by photon correlation spectroscopy and laser doppler anemometry. Further, no significant difference on biodistribution of poloxamer-preincubated liposomes was observed in comparison to control vesicles following intravenous administration into rats.

Many drugs of the future will be therapeutically active peptides and proteins developed through recombinant-DNA technology. A major factor limiting their exploitation is the current lack of appropriate non-parenteral delivery systems. Nasal systems incorporating absorption enhancers may provide a convenient, efficient means of administering protein and peptide therapeutics.

This paper describes an assessment of the potential of using bioadhesive microspheres as a nasal delivery system for biosynthetic human growth hormone (hGH) in sheep. The microsphere system was used alone and in combination with a biological surfactant, lysophosphatidylcholine (LPC). For comparison, hGH was also administered nasally as a solution and subcutaneously as an injection. The levels of hGH in the blood samples obtained were determined by an ELISA technique. The hGH was absorbed to only a very low extent when administered as a nasal solution. However, the microsphere delivery system without added enhancer was capable of considerably enhancing the nasal absorption of hGH. A delay in absorption was observed with the microspheres alone, which may be partially due to low aqueous solubility of hGH. Rapid and much higher absorption was observed when hGH was administered in combination with the microspheres and LPC as an enhancer.

The use of the bioadhesive microsphere system for the intranasal delivery of insulin was investigated in sheep. The effect of combining the above system with lysophosphatidylcholine, as a biological absorption enhancer, was also assessed. For comparison, insulin was also administered by the intravenous and subcutaneous routes. The plasma glucose and insulin levels were determined by a glucose oxidase method and a radioimmunoassay, respectively. The bioadhesive system produced large and significant changes in the plasma insulin and glucose levels. The relative bioavailability (± standard error of measurement) of insulin administered with the microsphere system alone was found to be 10.7% (±2.6) and that of insulin administered with the microsphere /enhancer system to be 31.5% (±6.5).

The effects of several prospective absorption enhancers were assessed on the nasal absorption of biosynthetic human growth hormone (hGH) in the rat. These enhancers function by alternative mechanisms that include enzyme inhibition, reduction in mucus viscosity, and enhancement of membrane fluidity. The levels of plasma hGH achieved were determined by an enzyme-linked immunosorbent assay. The increase in peak height was calculated relative to nasal administration of hGH alone without any enhancers and the relative bioavailability was calculated with reference to subcutaneous injection data. A lysophospholipid, lysophosphatidylcholine, gave the highest peak concentration, with an increase in peak height of 450% and a relative bioavailability of 25.8%. However, the greatest increase in AUC (291%) was achieved with the aminopeptidase inhibitor, amastatin, which gave a relative bioavailability of 28.9%. A mucolytic agent, N-acetyl-L-cysteine, and a transmembrane fatty acid transporter, palmitoyl-DL-carnitine, were also found to promote the nasal absorption of hGH in this model, with relative bioavailabilities of 12.2 and 22.1%, respectively. Bestatin, an enzyme inhibitor, was not an effective absorption enhancer for hGH in this model.

Different egg albumin microsphere systems for oral administration of paracetamol were prepared by the emulsion and capillary extrusion methods. The size of the microspheres depended on the method used to produce the microcapsules and also the size of the crystals of paracetamol. The effect of the following factors on in vitro dissolution were studied: different denaturation processes, variation in the ratio of paracetamol to albumin, size of microspheres, remnant oil in the microspheres, and the coating of the microspheres with membranes of polymeth-acrylates (Eudragit). The most important factors to control the release of paracetamol from the microspheres were the denaturation process and the use of waxes and membranes to delay the release of paracetamol from the microspheres. The egg albumin microspheres were very porous and permeable to water and thus the release of the paracetamol from the matrix was usually fast unless the microspheres were suitably coated, with, for example, Eudragit RL or RS.

Egg albumin microspheres containing paracetamol for oral administration were prepared and their in vivo characteristics evaluated. The egg albumin microspheres were able to improve the organoleptic characteristics of paracetamol formulations. The pharmacokinetic characteristics of three different formulations of paracetamol were evaluated in six volunteers. The formulations administered orally were: (1) Paracetamol granulated with lactose (reference), (2) Egg albumin microspheres and (3) Egg albumin microspheres coated with polymethacrylate. The pharmacokinetic characteristics for formulations 1 and 2 were similar but formulation 3 gave significant differences (p less than 0.05) in Ka, Cmax and tmax. No significant differences in relative bioavailability were observed.

The aim of this study was to investigate and quantify drug movement to the brain via the neuro-olfactory system after intranasal dosing of four model drugs; three glycine receptor antagonists and one angiotensin antagonist. The drugs were dosed to rats via intranasal or intravenous administration, after which a quantitative method for tissue distribution was utilised to determine drug distribution to the olfactory lobes, brain sections and the blood over 30 min. Autoradiography was used to visualise and quantify drug distribution throughout the brain and in the CSF. Micro-autoradiography was used to examine drug distribution throughout the olfactory nerve apparatus. The three glycine receptor antagonist compounds were transported to the CNS to differing degrees although they had similar molecular structures and similar physicochemical characteristics. All three compounds were shown to exploit a direct route of transport from nose to brain with Direct Transport Percentages (DTP) of 99.99%, 96.71% and 51.95%, respectively, although for the last molecule a major part of the brain content originated from systemic transport across the BBB. Intranasal administration of GR138950 resulted in over 3.5 times more drug in the olfactory lobes at 1 min post-dose compared to intravenous administration; and 5 times more drug was delivered to the olfactory lobes over 30 min. Micro-autoradiography showed that GR138950 could be found throughout the olfactory nerve apparatus. Autoradiography illustrated drug distribution throughout the brain and CSF, with drug concentrations in the CSF being equal or higher than in the brain tissue. It was determined that approximately 0.8% of the administered dose moved into the brain and CSF via the olfactory pathway over 30 min. Intranasal administration resulted in greater delivery of the model drugs to the olfactory lobes and brain as compared to intravenous dosing. It is proposed that the drug moved through the neuro-olfactory system, primarily via paracellular pathways.

It has been shown that vasoconstrictive drugs such as ephedrine derivatives are able to decrease systemic absorption of drugs administered by mucosal surfaces. The present paper set out to evaluate in the rat model the effect of co-administered nasal ephedrine on the absorption of GR138950 in a simple and in a pectin self-gelling formulation. It was hypothetised that a decrease in nasal systemic absorption would lead to an increase in direct nose-to-brain transport as demonstrated by the drug concentration in the olfactory lobes of the brain. It was found that ephedrine administered nasally with the drug in a simple aqueous solution resulted in a significant increase in nasal systemic absorption and also an increase in brain delivery; however, this trend was not observed with the pectin formulations. The pectin formulation with ephedrine resulted in lower systemic absorption of GR138950 and lower brain uptake compared to the simple solution formulation containing ephedrine.

Interest in intranasal (IN) administration as a non-invasive route for drug delivery continues to grow rapidly. The nasal mucosa offers numerous benefits as a target issue for drug delivery, such as a large surface area for delivery, rapid drug onset, potential for central nervous system delivery, and no first-pass metabolism. A wide variety of therapeutic compounds can be delivered IN, including relatively large molecules such as peptides and proteins, particularly in the presence of permeation enhancers. The current review provides an in-depth discussion of therapeutic aspects of IN delivery including consideration of the intended indication, regimen, and patient population, as well as physicochemical properties of the drug itself. Case examples are provided to illustrate the utility of IN dosing. It is anticipated that the present review will prove useful for formulation scientists considering IN delivery as a delivery route.

This review discusses the possible benefits of using nanoparticles for nasal delivery of drugs and vaccines. It considers the various factors affecting particle transport across the nasal tissue. The evidence for the improved transport of drugs, such as peptides and proteins, across the nasal epithelium when formulated in a nanoparticulate system, as compared to an optimal solution formulation, is not convincing. For instance it has been shown that a chitosan solution and especially a chitosan powder formulation was superior in enhancing the nasal absorption of insulin as compared to chitosan nanoparticles. On the other hand, the use of nanoparticles for vaccine delivery seems beneficial in that good immune responses are achieved. This could be due to the fact that small particles can be transported preferentially by the lymphoid tissue of the nasal cavity (NALT). However, apparently no studies have been published comparing directly other adjuvant nasal systems with nanoparticulate systems.

There is an increasing need to identify novel approaches by which to improve the efficiency of drug transport from the nasal cavity (olfactory region) to the CNS, especially for treatment of central nervous system disorders. It is suggested, that one approach is the combination of active targeting of a bioadhesive formulation, that will retain the drug at the absorption site, potentially in combination with, an absorption enhancer. Two low methylated pectins, LM-5 and LM-12 were selected for evaluation as drug delivery systems, due to their ability to gel in the nasal cavity and their bioadhesive characteristics, together with chitosan G210, which acts both as a bioadhesive material and as an efficient absorption enhancer. It was found that all of the bioadhesive formulations were able to reach the olfactory region in the nasal cavity of human volunteers when delivered using a simple nasal drop device. Furthermore, the formulations displayed a significantly increased residence time on the epithelial surface. This was in contrast to a non-bioadhesive control delivered with the same device. In contrast, a pectin formulation administered with a nasal spray system did not show an increase in residence time in the olfactory region. It was further shown that the reproducibility of olfactory delivery of a polymer formulation was significantly better intra-subject than inter-subject.

There is an increasing need for nasal drug delivery systems that could improve the efficiency of the direct nose to brain pathway especially for drugs for treatment of central nervous system disorders. Novel approaches that are able to combine active targeting of a formulation to the olfactory region with controlled release bioadhesive characteristics, for maintaining the drug on the absorption site are suggested. If necessary an absorption enhancer could be incorporated. Low methylated pectins have been shown to gel and be retained in the nasal cavity after deposition. Chitosan is known to be bioadhesive and also to work as an absorption enhancer. Consequently, two types of pectins, LM-5 and LM-12, together with chitosan G210, were selected for characterisation in terms of molecular weight, gelling ability and viscosity. Furthermore, studies on the in vitro release of model drugs from candidate formulations and the transport of drugs across MDCK1 cell monolayers in the presence of pectin and chitosan were also performed. Bioadhesive formulations providing controlled release with increased or decreased epithelial transport were developed. Due to their promising characteristics 3% LM-5, 1% LM-12 pectin and 1% chitosan G210 formulations were selected for further biological evaluation in animal models.

This paper reviews the anatomical and physiological factors of importance for nasal drug delivery and discusses in particular the influence of the nasal mucociliary clearance mechanism on the nasal absorption of drugs. The effect of nasal pathological conditions on the mucociliary clearance mechanism and the possible effect of such disease states on nasal drug transport are also discussed. Strategies for the exploitation of bioadhesive drug delivery systems and especially nasal absorption enhancers for the improvement of nasal drug delivery are evaluated to include considerations of the mechanism of action and correlation between the degree of bioadhesion and absorption enhancement and transport of drugs across the nasal membrane. A range of studies involving bioadhesive/absorption enhancer systems are detailed. A selected bioadhesive material, chitosan, which has been shown to have excellent absorption enhancer properties for a variety of drugs is discussed in some detail.

The adsorption behaviour of a tetrafunctional copolymer of poly (ethylene oxide)-poly (propylene oxide) ethylene diamine (commercially available as Poloxamine 908) and a diblock copolymer of poly (lactic acid)-poly (ethylene oxide) (PLA/PEG 2:5) onto a model colloidal drug carrier (156 nm sized polystyrene latex) is described. The adsorption isotherm, hydrodynamic thickness of the adsorbed layers and enthalpy of the adsorption were assessed. The close similarity in the conformation of the poly (ethylene oxide) (PEO) chains (molecular weight 5,000 Da) in the adsorbed layers of these two copolymers was demonstrated by combining the adsorption data with the adsorbed layer thickness data. In contrast, the results from isothermal titration microcalorimetry indicated a distinct difference in the interaction of the copolymers with the polystyrene colloid surface. Poloxamine 908 adsorption to polystyrene nanoparticles is dominated by an endothermic heat effect, whereas, PLA/PEG 2:5 adsorption is entirely an exothermic process. This difference in adsorption behaviour could provide an explanation for differences in the biodistribution of Poloxamine 908 and PLA/PEG 2:5 coated polystyrene nanoparticles observed in previous studies. A comparison with the interaction enthalpy for several other PEO-containing copolymers onto the same polystyrene colloid was made. The results demonstrate the importance of the nature of the anchoring moiety on the interaction of the adsorbing copolymer with the colloid surface. An endothermic contribution is found when an adsorbing molecule contains a poly (propylene oxide) (PPO) moiety (e.g. Poloxamine 908), whilst the adsorption is exothermic (i.e. enthalpy driven) for PEO copolymers with polylactide (PLA/PEG 2:5) or alkyl moieties.

Nasal influenza vaccination may prove to be a good alternative to parenteral injection because of the enhancement of the mucosal immune response and the ease of vaccine administration. This study investigated the use of chitosan, a bioadhesive polymer, as a nasal delivery system with inactivated, subunit influenza vaccine. Subjects received nasally 15 or 7.5 microg of the standard inactivated trivalent influenza vaccine with chitosan or 15 microg of the same vaccine intramuscularly. Serum haemagglutination inhibition (HI) titres for all three vaccine components were measured prior to, and at time points up to 14 weeks after dosing. Serum HI titres following intranasal vaccination with the nasal chitosan-influenza vaccine met the criteria set by the Committee for Proprietary Medicinal Products in terms of seroprotection rate, seroconversion rate and mean fold increase of HI titre for at least one of the three antigens in the vaccination schedules used. These data show that nasal immunisation with chitosan plus trivalent inactivated influenza is a potentially effective, easily-administered form of vaccination.

Respiratory syncytial virus (RSV), an important pathogen of the lower respiratory tract, is responsible for severe illness both in new born and young children and in elderly people. Due to complications associated with the use of the early developed vaccines, there is still a need for an effective vaccine against RSV. Most pathogens enter the body via mucosal surfaces and therefore vaccine delivery via routes such as the nasal, may well prove to be superior in inducing protective immune responses against respiratory viruses, since both local and systemic immunity can be induced by nasal immunisation. Previously we have shown that intradermal immunisation of a plasmid DNA encoding the CTL epitope from the M2 protein of RSV induced protective CTL responses. In the present study, the mucosal delivery of plasmid DNA formulated with chitosan has been investigated. Chitosan is a polysachharide consisting of copolymers of N-acetylglucosamine and glucosamine that is derived from chitin, a material found in the shells of crustacea. Intranasal immunisation with plasmid DNA formulated with chitosan induced peptide- and virus-specific CTL responses in BALB/c mice that were comparable to those induced via intradermal immunisation. Following RSV challenge of chitosan/DNA immunised mice, a significant reduction (P

Subunit intranasal vaccines offer the prospect of inducing combined systemic-mucosal immunity against mucosally transmitted infections such as human immunodeficiency virus. However, although human studies have demonstrated the induction of active immunity, secretory immunoglobulin A (sIgA) responses are variable, and no study has demonstrated protection by accepted vaccine-licensing criteria as measured by direct toxin-neutralizing activity. Using the genetically inactivated mutant diphtheria toxoid CRM 197 in a bioadhesive polycationic polysaccharide chitosan delivery system, we found that a single nasal immunization was well tolerated and boosted antitoxin neutralizing activity in healthy volunteers, which could be further boosted by a second immunization. The neutralizing activity far exceeded accepted protective levels and was equivalent to that induced by standard intramuscular vaccine and significantly greater than intranasal immunization with CRM 197 in the absence of chitosan. A striking but unexpected observation was that although unilateral intranasal immunization induced circulating antitoxin antibody-secreting cells, a nasal antitoxin sIgA response was seen only after the second immunization and only in the vaccinated nostril. If these data are reproduced in larger studies, an intranasal diphtheria vaccine based on CRM 197 -chitosan could be rapidly licensed for human use. However, a restricted sIgA response suggests that care must be taken in the priming-boosting strategy and clinical sampling techniques when evaluating such vaccines for the induction of local mucosal immunity.

A novel nasal formulation, in the form of a nicotine-Amberlite resin complex powder has been developed that provided an optimal combined pulsatile and sustained plasma nicotine profile for smoking cessation. The adsorption isotherms of nicotine hydrogen tartrate salt on two types of Amberlite resins (IRP69 and IR120) were evaluated and the subsequent in vitro release properties of nicotine from the nicotine-Amberlite complex powders were tested using a Franz diffusion cell. Amberlite IRP69 and Amberlite IR120 are similar cationic exchange materials with the same ion-exchange capacity but due to a smaller particle size range (10-150 microm) Amberlite IRP69 had a better flow property and a better adsorptive capacity than Amberlite IR120. The material is used as an excipient in marketed pharmaceutical formulations. The highly water soluble salt, nicotine hydrogen tartrate, displayed good adsorption onto both types of Amberlite resin. The maximum adsorption of nicotine onto Amberlite IRP69 was 1.071 mg drug per mg resin. The cumulative release of drug from nicotine hydrogen tartrate-Amberlite complex powders showed that the higher the drug loading, the faster was the rate of release of the drug. Based on these results, various nicotine hydrogen tartrate-Amberlite IRP69 powder formulations containing different ratios of free to bound drug (50% to 100% bound) and a control solution were prepared and evaluated in a sheep model by nasal administration. The nicotine plasma profiles demonstrated that an initial rapid peak plasma level of nicotine followed by a sustained elevated level could be achieved by adjusting the ratio of free to bound nicotine in the Amberlite powder formulation. The curves obtained from some of the formulations were comparable to those predicted from a computer-generated pharmacokinetic model.

In recent years, several strategies have been under investigation to achieve safe and effective immunisation, in terms of new antigens, adjuvants and routes of vaccination. The latter include mucosal sites such as oral, rectal, vaginal and nasal. Biodegradable microparticles produced from polymers such as poly(D,L-lactide) (PLA) and poly(D,L-lactide-co-glycolide) (PLGA) containing encapsulated vaccine antigens have been extensively studied for immunisation. These microparticles allow controlled release of vaccines with the aim to develop as single dose vaccines. However there are concerns regarding the integrity and immunogenicity of the antigen during the encapsulation process when the antigen is exposed to organic solvents, high shear stresses and the exposure of antigen to low pH which is caused by polymer degradation. Polymeric lamellar substrate particles (PLSP) produced by simple precipitation of PLA, form a novel polymeric system for the adsorption of antigens. This procedure avoids pH changes, exposure to organic solvents and hence allows the integrity of the antigen to be retained. The aim of this article is to discuss the factors affecting the characteristics of PLSP and adsorption of antigens onto PLSP and consider their potential as adjuvants for the nasal delivery of protein, peptide or viral vaccines.

While covalent attachment of small drug molecules to AB copolymers for the formation of polymeric micelles for drug delivery has been investigated, few studies have focused on non-covalent interactions. The aim of this study was therefore to explore the potential of non-covalent interactions between an AB copolymer, Poly(aspartic acid)–poly(ethylene glycol) (Pasp–PEG), with anionic pendant groups and diminazene aceturate, a small molecular weight cationic drug. Micelles were prepared by mixing solutions of Pasp–PEG and diminazene in 25 mM Tris–HCl buffer. At all Pasp–PEG concentrations studied, the micelles appeared to be water soluble with a unimodal size distribution and ranged in size from approximately 22 to 60 nm. The polyionic micelles also displayed similar and small absolute zeta potential values at various drug:monomer molar ratios which confirmed stabilisation by the PEG corona. The scattering intensity was maximal and remained unchanged, while particle size increased slightly at pH range from 3.4 to 7.2. At this pH range both the polymer and drug would be ionised and ionic interactions possible to drive micellar formation. An increase in size and scattering intensity with addition of NaCl to the micelles was attributed to dehydration of the PEG corona which may have led to aggregation of the micelles. The absence of micellar dissociation upon addition of salt was attributed to the dominance of hydrogen bonding between Pasp and diminazene aceturate, as assessed by isothermal titration microcalorimetry. Morphological evaluation of these constructs showed them to be discrete and fairly uniform in size and shape. This study was therefore successful in confirming the potential of non-covalent interactions using an AB copolymer to form polyionic micelles for drug delivery.

This paper investigates the effect of starch microspheres on the absorption enhancing efficiency of various enhancer systems in formulations with insulin after application in the nasal cavity of sheep. The enhancers studied were lysophosphatidylcholine, glycodeoxycholate and sodium taurodihydroxyfusidate, a bile salt derivative. The enhancers were selected on the basis of their perceived or proven mechanism of action and worked predominantly by interacting with the lipid membrane. The bioadhesive starch microspheres were shown to increase synergistically the effect of the absorption enhancers on the transport of the insulin across the nasal membrane. Dependent on the potency of the enhancer system the increment in absorption enhancement was shown to be from 1.4 times to 5 times that obtained for the absorption enhancer in solution.

The nasal cycle is a well-recognised physiological phenomenon where each side of the nose alternates through phases of congestion and decongestion. Although many physiological properties of the nose alternate with the nasal cycle whether this has any effect on the nasal mucociliary clearance is less clear. As the nose is a potential site for the administration of pharmaceuticals, it is essential that any factors that could affect clearance (and hence absorption) are identified. This study set out to investigate if mucociliary clearance rates differed between the clear and obstructed airway at a morning peak of the nasal cycle in five healthy volunteers with normal nasal anatomy using a dual-radioisotope labelling procedure that allows both sides of the nose to be assessed simultaneously. The clearance of the radiopharmaceutical formulations from the nasal cavity was monitored using gamma scintigraphy and decay-adjusted 50%-clearance times were calculated for each nostril. The ratios of clearance times from the patent nostril when compared to the obstructed nostril were statistically significant (two-tailed t-test; P = 0.039), the mean ratio being 2.5 : 1 (SEM +/- 0.5). It can be concluded that the nasal cycle has a marked effect on the mucociliary clearance patterns of the nose. This may have both theoretical and practical implications for the nasal delivery of drugs.

Surface-modified albumin nanoparticles were prepared from two poly(ethylene glycol)-human serum albumin conjugates: poly(thioetheramido acid)-poly(ethylene glycol) copolymer-grafted HSA (HSA-PTAAC-PEG) and methoxy poly(ethylene glycol)-grafted HSA (HSA-mPEG). Rose bengal (RB) was used as a model drug for encapsulation into the nanoparticles either during the particle production or by adsorption post particle preparation. The drug incorporation and release was affected by the different production methods and the different polymer compositions. When RB was loaded in HSA and HSA/HSA-PTAAC-PEG nanoparticles, up to 5% (w/w) drug content was achieved. The drug loading in HSA-mPEG nanoparticles was much lower and the results from the microcalorimetry study indicated that the low loading efficiency was due to less drug-protein binding sites available in the HSA-mPEG molecule as compared to the HSA molecule. The release of RB from the albumin nanoparticles was very slow in PBS and dramatically accelerated in the presence of trypsin. Compared with unmodified nanoparticles, the slower release of RB from the surface-modified HSA nanoparticles in the presence of the enzyme suggested that the existence of a steric hydrophilic barrier on the surface of the nanoparticles made digestion of the nanoparticles more difficult.

Self-consistent field (SCF) modelling studies on a series of varying molecular weight poly(lactic acid):poly(ethylene glycol) (PLA:PEG) particles (2:5–15:5 kDa) that form micelles in aqueous solutions have been performed using the formalism of Scheutjens and Fleer. The parameters were chosen to represent a central core made from collapsed hydrophobic PLA blocks, and PEG blocks, which form a corona layer extended into the solvent. These different phases have been confirmed in 1 H and 13 C NMR experiments. From the analysis of the relevant thermodynamic data of the series, particle radii and aggregation numbers varying from 12.6 to 24.9 nm and from 17 to 280, respectively, have been predicted, which agree well with those obtained experimentally. The actual thickness of the PEG corona is fairly constant at approximately 11 nm for the whole particle series indicating that the size of the PLA core has little effect on the thickness of the PEG corona layer. Another important consideration of these particles is the PEG density at the surface, which is related to the in vivo properties of these particles. The surface area available to each PEG block at the central PLA core has been obtained in order to compare the members of the particle series. It has been shown that the surface area available to each PEG block decreases and then increases throughout the PLA:PEG series.

We have evaluated the ability of two carbohydrate biopolymers, chitosan and gellan, to enhance antibody responses to subunit influenza virus vaccines delivered to the respiratory tracts of mice. Groups of mice were vaccinated three times intranasally (i.n.) with 10 μg of purified influenza B/Panama virus surface antigens (PSAs), which consist of hemagglutinin (HA) and neuraminidase (NA), either alone or admixed with chitosan or gellan solutions. Separate groups were vaccinated subcutaneously (s.c.) with PSAs adsorbed to Alhydrogel or chitosan or gellan alone i.n. Serum antibody responses were determined by enzyme-linked immunosorbent assay (ELISA) for influenza virus-specific immunoglobulin G (IgG) and by HA inhibition (HAI) and NA inhibition (NAI) assays. The local respiratory immune response was measured by assaying for influenza virus-specific IgA antibody in nasal secretions and by enumerating nasal and pulmonary lymphocytes secreting IgA, IgG, and IgM anti-influenza virus-specific antibodies by enzyme-linked immunospotting (ELISPOT). When administered alone i.n., B/Panama PSA was poorly immunogenic. Parenteral immunization with B/Panama PSA with Alhydrogel elicited high titers of anti-B/Panama antibodies in serum but a very poor respiratory anti-B/Panama IgA response. In contrast, i.n. immunization with PSA plus chitosan stimulated very strong local and systemic anti-B/Panama responses. Gellan also enhanced the local and serum antibody responses to i.n. PSA but not to the same extent as chitosan. The ability of chitosan to augment the immunogenicity of influenza vaccines given i.n. was confirmed using PSA prepared from an influenza A virus (A/Texas H1N1).

Schizophrenia is a severe non-curable illness of the brain with serious consequences if not properly treated and kept under control. Antipsychotic drugs have revolutionised the therapy and management of schizophrenia. However, patient compliance rates are notoriously poor due to the nature of the disease and troublesome side-effects, and are major causes of symptom recurrence. Although some new antipsychotic agents have been marketed to offer broader efficacy with much reduced side-effect profiles, the drug delivery systems for antipsychotics are still in the stage of conventional dosage forms, such as tablets, capsules and solutions, and need to be dosed at the frequency of 2-4 times daily. Doubtless. novel drug delivery systems, such as sustained and controlled release systems, will be useful for antipsychotics. They should reduce the frequency of dosing. enhance drug bioavailability and improve patient compliance. In this article, the specificity and characterisation of schizophrenia and pathophysiology. drug therapy. and the development and future prospects of neuroleptic drug delivery systems are reviewed.