Increasing Cellular Uptake and Permeation of Curcumin Using a Novel Polymer-Surfactant Formulation.

Several therapeutically active molecules are poorly water-soluble, thereby creating a challenge for pharmaceutical scientists to develop an active solution for their oral drug delivery. This study aimed to investigate the potential for novel polymer-surfactant-based formulations (designated A and B) to improve the solubility and permeability of curcumin. A solubility study and characterization studies (FTIR, DSC and XRD) were conducted for the various formulations. The cytotoxicity of formulations and commercial comparators was tested via MTT and LDH assays, and their permeability by in vitro drug transport and cellular drug uptake was established using the Caco-2 cell model. The apparent permeability coefficients (Papp) are considered a good indicator of drug permeation. However, it can be argued that the magnitude of Papp, when used to reflect the permeability of the cells to the drug, can be influenced by the initial drug concentration (C0) in the donor chamber. Therefore, Papp (suspension) and Papp (solution) were calculated based on the different values of C0. It was clear that Papp (solution) can more accurately reflect drug permeation than Papp (suspension). Formulation A, containing Soluplus® and vitamin E TPGs, significantly increased the permeation and cellular uptake of curcumin compared to other samples, which is believed to be related to the increased aqueous solubility of the drug in this formulation.

Design considerations for future radionuclide aerosol monitoring systems.

Pacific Northwest National Laboratory (PNNL) staff developed the Radionuclide Aerosol Sampler Analyzer (RASA) for worldwide aerosol monitoring in the 1990s. Recently, researchers at PNNL and Creare, LLC, have investigated possibilities for how RASA could be improved, based on lessons learned from more than 15 years of continuous operation, including during the Fukushima Daiichi Nuclear Power Plant disaster. Key themes addressed in upgrade possibilities include having a modular approach to additional radionuclide measurements, optimizing the sampling/analyzing times to improve detection location capabilities, and reducing power consumption by using electrostatic collection versus classic filtration collection. These individual efforts have been made in a modular context that might constitute retrofits to the existing RASA, modular components that could improve a manual monitoring approach, or a completely new RASA. Substantial optimization of the detection and location capabilities of an aerosol network is possible and new missions could be addressed by including additional measurements.

Using Atmospheric Dispersion Theory to Inform the Design of a Short-lived Radioactive Particle Release Experiment.

Atmospheric dispersion theory can be used to predict ground deposition of particulates downwind of a radionuclide release. This paper uses standard formulations found in Gaussian plume models to inform the design of an experimental release of short-lived radioactive particles into the atmosphere. Specifically, a source depletion algorithm is used to determine the optimum particle size and release height that maximizes the near-field deposition while minimizing both the required source activity and the fraction of activity lost to long-distance transport. The purpose of the release is to provide a realistic deposition pattern that might be observed downwind of a small-scale vent from an underground nuclear explosion. The deposition field will be used, in part, to study several techniques of gamma radiation survey and spectrometry that could be used by an On-Site Inspection team investigating such an event.

PRex: An Experiment to Investigate Detection of Near-field Particulate Deposition from a Simulated Underground Nuclear Weapons Test Vent.

A radioactive particulate release experiment to produce a near-field ground deposition representative of small-scale venting from an underground nuclear test was conducted to gather data in support of treaty capability development activities. For this experiment, a CO2-driven "air cannon" was used to inject (140)La, a radioisotope of lanthanum with 1.7-d half-life and strong gamma-ray emissions, into the lowest levels of the atmosphere at ambient temperatures. Witness plates and air samplers were laid out in an irregular grid covering the area where the plume was anticipated to deposit based on climatological wind records. This experiment was performed at the Nevada National Security Site, where existing infrastructure, radiological procedures, and support personnel facilitated planning and execution of the work. A vehicle-mounted NaI(Tl) spectrometer and a polyvinyl toluene-based backpack instrument were used to survey the deposited plume. Hand-held instruments, including NaI(Tl) and lanthanum bromide scintillators and high purity germanium spectrometers, were used to take in situ measurements. Additionally, three soil sampling techniques were investigated and compared. The relative sensitivity and utility of sampling and survey methods are discussed in the context of on-site inspection.

An evaluation of the adhesion of solid oral dosage form coatings to the oesophagus.

There is a requirement for the development of oral dosage forms that are adhesive and allow extended oesophageal residence time for localised therapies, or are non-adhesive for ease of swallowing. This study provides an initial assessment of the in vitro oesophageal retention characteristics of several widely utilised pharmaceutical coating materials. To this end, a previously described apparatus has been used to measure the force required to pull a coated disc-shaped model tablet across a section of excised oesophageal tissue. Of the materials tested, the well-studied mucoadhesive polymer sodium alginate was found to be associated with significant oesophageal adhesion properties that was capable of 'self-repairing'. Hydroxypropylmethylcellulose exhibited less pronounced bioadhesive behaviour and blending this with plasticiser or with low molecular weight polymers and surfactants did not significantly affect this. Low molecular weight water soluble polymers, were found to behave similarly to the uncoated glass control disc. Polysorbates exhibited bioadhesion behaviour that was majorly influenced by the nature of the surfactant. The insoluble polymer ethylcellulose, and the relatively lipophilic surfactant sorbitan monooleate were seen to move more readily than the uncoated disc, suggesting that these may have a role as 'easy-to-swallow' coatings.

Delivery of treatment for hepatitis C virus infection in the primary care setting.

OBJECTIVES: The aim of this study was to evaluate the feasibility, safety and efficacy of treatment for chronic hepatitis C virus (HCV) infection through a primary care-based model for the delivery of HCV services in New South Wales (NSW), Australia. PARTICIPANTS AND METHODS: This observational cohort study recruited participants through seven primary care clinics in NSW, Australia, between November 2010 and June 2013. Patients with HCV genotype 2/3 were treated without specialist review, whereas those with genotype 1 required an initial specialist review. Treatment consisted of pegylated interferon-α-2a/2b and ribavirin. Sustained virological response and adverse events were evaluated. RESULTS: Among 41 participants (mean age 44 years, 73% men) initiating treatment with pegylated interferon-α-2a/2b and ribavirin, 90% had injected drugs ever, 16% had injected drugs in the past 30 days and 56% had ever received opioid substitution treatment. HCV genotype 1 and genotype 2/3 occurred in 17% (n=7) and 83% (n=34). Treatment was completed in 83% (34 of 41), with seven discontinuations [adverse event (depression), n=1; patient decision, n=1; lost to follow-up, n=3; virological nonresponse, n=2]. In an intent-to-treat analysis, sustained virological response was 71% overall (29 of 41), 43% in genotype 1 (three of seven) and 76% in genotype 2/3 (26 of 34). CONCLUSION: Initiation of HCV treatment in the primary care setting is an effective alternative for selected patients and may contribute towards increasing access to HCV care.

An in vitro model for the evaluation of the adhesion of solid oral dosage forms to the oesophagus.

Adhesion of solid oral dosage forms to the oesophagus can be a disadvantage when delivering drugs that may cause oesophageal damage, or can be an advantage when developing localised therapies for this region. In this study, apparatus to investigate coatings that may influence oesophageal retention was developed and evaluated. The apparatus incorporated a section of porcine oesophageal mucosa held flat by the application of a gentle vacuum and kept moist by the application of a simulated saliva solution. The resistance to the application of more physiologically relevant shear stresses was evaluated. Using a range of materials it was found that differences in oesophageal adhesion could be identified. Materials like sodium alginate were highly adhesive and had a tendency to re-adhere while paraffin waxes showed no adhesion. The rapid loss of the polymer coat from the surface for water swellable materials was identified as an issue.

Nasal and buccal drug delivery: management forum conference.

The scope of the conference (Nasal and Buccal Drug Delivery Conference, Management Forum; Chairs Franz Merkus and Julie Suman) was to consider innovations in drug delivery via the nose and oral cavity, notably for the delivery of vaccines, antimalarials and rapidly acting sedatives. Presentations from experts from academia, government agencies and commercial organisations were made over the 2 days. The advantages of both routes were ease of application, patient acceptability and no requirement to produce sterile products. These routes worked best for drugs that are water soluble--but with some lipophilicity--only require low doses, are acceptable to the patient and have low irritancy (particulary for the nasal route). Challenges relate to the effectiveness of deposition from the delivery systems and the efficient clearance mechanisms. It was concluded that for many drugs, buccal and nasal delivery could become the route of choice for their application; vaccines, in particular, appear to show promise for nasal delivery.

Chitosan microparticles for the controlled delivery of fluoride.

OBJECTIVES: To manufacture and characterise chitosan/fluoride microparticles prepared by spray drying and assess their utility as controlled release vehicles for fluoride. METHODS: Microparticles were manufactured from dispersions containing 1.0% and 2.0% (w/v) chitosan and 0.20% or 0.40% (w/v) NaF in the absence/presence of glutaraldehyde. Particle size distributions were determined using laser diffraction; fluoride loading and release were determined by ion-selective electrode. Release profiles were studied in isotonic media (pH 5.5) over 360 min; microparticles exhibiting greatest cumulative fluoride release were further evaluated at pH 4.0 and 7.0. Particle morphology was investigated using environmental scanning electron microscopy. Bioadhesion parameters were determined with a texture-probe analyser. RESULTS: Microparticles exhibited low polydispersity and volume mean diameters (VMDs) <6 µm. VMDs increased on doubling the chitosan/fluoride concentrations but were largely independent of glutaraldehyde concentration. Recovered yields were inversely proportional to dispersion viscosity due to compromised fluid atomisation; adding NaF reduced viscosity and improved yields. Best-case entrapment efficiency and NaF loading were 84.1% and 14%, respectively. Release profiles were biphasic, releasing 40-60% of the total fluoride during the first 600 s, followed by a prolonged release phase extending out to 6h. Incorporation of 0.40% NaF to the 2.0% chitosan dispersion yielded microparticles with reduced bioadhesive parameters (F(max) and WOA) versus the chitosan-only control whilst retaining significant bioadhesive potential. CONCLUSIONS: Bioadhesive chitosan/fluoride microparticles manufactured using a spray-drying protocol have been extensively characterised and further opportunity for optimisation identified. These microparticles may provide a means of increasing fluoride uptake from oral care products to provide increased protection against caries, however further work is required to demonstrate this principle in vivo. CLINICAL SIGNIFICANCE: Spray-drying is a low-cost route for the manufacture of bioadhesive chitosan/fluoride microparticles which can be exploited as controlled fluoride release agents to aid fluoride retention in the oral cavity. The potential exists to optimise release profiles to suit the delivery format thereby maximising the cariostatic benefits.

Metabolism of captopril carboxyl ester derivatives for percutaneous absorption.

OBJECTIVES: To determine the metabolism of captopril n-carboxyl derivatives and how this may impact on their use as transdermal prodrugs. The pharmacological activity of the ester derivatives was also characterised in order to compare the angiotensin converting enzyme inhibitory potency of the derivatives compared with the parent drug, captopril. METHODS: The metabolism rates of the ester derivatives were determined in vitro (using porcine liver esterase and porcine ear skin) and in silico (using molecular modelling to investigate the potential to predict metabolism). KEY FINDINGS: Relatively slow pseudo first-order metabolism of the prodrugs was observed, with the ethyl ester displaying the highest rate of metabolism. A strong relationship was established between in-vitro methods, while in-silico methods support the use of in-vitro methods and highlight the potential of in-silico techniques to predict metabolism. All the prodrugs behaved as angiotensin converting enzyme inhibitors, with the methyl ester displaying optimum inhibition. CONCLUSIONS: In-vitro porcine liver esterase metabolism rates inform in-vitro skin rates well, and in-silico interaction energies relate well to both. Thus, in-silico methods may be developed that include interaction energies to predict metabolism rates.

Clinical relevance of dissolution testing in quality by design.

Quality by design (QbD) has recently been introduced in pharmaceutical product development in a regulatory context and the process of implementing such concepts in the drug approval process is presently on-going. This has the potential to allow for a more flexible regulatory approach based on understanding and optimisation of how design of a product and its manufacturing process may affect product quality. Thus, adding restrictions to manufacturing beyond what can be motivated by clinical quality brings no benefits but only additional costs. This leads to a challenge for biopharmaceutical scientists to link clinical product performance to critical manufacturing attributes. In vitro dissolution testing is clearly a key tool for this purpose and the present bioequivalence guidelines and biopharmaceutical classification system (BCS) provides a platform for regulatory applications of in vitro dissolution as a marker for consistency in clinical outcomes. However, the application of these concepts might need to be further developed in the context of QbD to take advantage of the higher level of understanding that is implied and displayed in regulatory documentation utilising QbD concepts. Aspects that should be considered include identification of rate limiting steps in the absorption process that can be linked to pharmacokinetic variables and used for prediction of bioavailability variables, in vivo relevance of in vitro dissolution test conditions and performance/interpretation of specific bioavailability studies on critical formulation/process variables. This article will give some examples and suggestions how clinical relevance of dissolution testing can be achieved in the context of QbD derived from a specific case study for a BCS II compound.

Azo compounds in colon-specific drug delivery.

Azo compounds have the potential to act as drug carriers that facilitate the selective release of therapeutic agents to the colon, and also to effect the oral administration of those macromolecular drugs that require colon-specific drug delivery. With some further research-driven refinements, these materials may lead to more efficient treatments for local conditions, such as colonic cancer or inflammatory bowel disease. This article provides an overview of the azo-based systems developed to date, identifies the requirements for an ideal carrier, and highlights the directions for further developments in the field of azo group-facilitated colonic delivery.

An in vitro assessment of bioadhesive zinc/carbomer complexes for antimicrobial therapy within the oral cavity.

The effectiveness of antimicrobial agents in the oral cavity is limited by their retention at the site of action. In this work the antimicrobial cation zinc was complexed with the bioadhesive agent Carbopol 971P, in order to allow an extended antimicrobial effect. Zinc ions were shown to form stable complexes with the polymer, and were not released into distilled water. However, in the presence of other cations, it was possible to displace zinc over an extended period. A low pH was seen to enhance zinc release. The complexes were found to have similar bioadhesive properties to the polymer alone when tested using a buccal cell adsorption model and texture probe analysis. It was concluded that this complex shows promise as a means of allowing the extended delivery of zinc ions locally within the oral cavity.

Orally administered, colon-specific mucoadhesive azopolymer particles for the treatment of inflammatory bowel disease: An in vivo study.

Radiolabeled congeners of a series of azopolymers have been synthesized and characterized. The in vivo (rat) gastrointestinal transit profile of millimeter-sized particles of these azopolymers has been determined and used to facilitate the selection of a candidate material for therapeutic applications. The efficacy of the selected material as a protective coating for the colonic mucosa has been tested in a hapten-reactivated, in vivo model of inflammatory bowel disease: 7 days after reactivation of the condition, the myeloperoxidase activity of animals that had received doses of the selected azopolymer was determined to be at the same level as that of healthy animals or that of the negative control group, highlighting the therapeutic promise of this material.

Design, synthesis and characterization of captopril prodrugs for enhanced percutaneous absorption.

Most drugs are designed primarily for oral administration, but the activity and stability profiles desirable for this route often make them unsuitable for transdermal delivery. We were therefore interested in designing analogues of captopril, a model drug with poor percutaneous penetration, for which the sustained steady-state blood plasma level associated with transdermal delivery (and which is unattainable orally) would be particularly beneficial. Quantitative structure-permeability relationships (QSPRs) predicted that ester and thiol prodrug derivatives of captopril would have lower maximal transdermal flux (J(m)) than the parent drug, since the increases in permeability coefficient (k(p)) of prodrugs would be outweighed by the reductions in aqueous solubility. Therefore, the aim of this study was to synthesize a series of prodrugs of captopril and to determine if a QSPR model could be used to design therapeutically viable prodrugs. Molecules with the highest predicted k(p) values were synthesized and characterized, and J(m) measured in Franz diffusion cells from saturated aqueous donor across porcine skin (fresh and frozen). In-vitro metabolism was also measured. Captopril and the prodrugs crossed the skin relatively freely, with J(m) being highest for ethyl to butyl esters. Substantial first-order metabolism of the prodrugs was observed, suggesting that their enhanced percutaneous absorption was complemented by their metabolic performance. The results suggested that QSPR models provided excellent enhancements in drug delivery. This was not seen at higher lipophilicities, suggesting that issues of solubility need to be considered in conjunction with any such use of a QSPR model.

Buccal drug delivery.

Buccal formulations have been developed to allow prolonged localised therapy and enhanced systemic delivery. The buccal mucosa, however, while avoiding first-pass effects, is a formidable barrier to drug absorption, especially for biopharmaceutical products (proteins and oligonucleotides) arising from the recent advances in genomics and proteomics. The buccal route is typically used for extended drug delivery, so formulations that can be attached to the buccal mucosa are favoured. The bioadhesive polymers used in buccal drug delivery to retain a formulation are typically hydrophilic macro-molecules containing numerous hydrogen bonding groups. Newer second-generation bioadhesives have been developed and these include modified or new polymers that allow enhanced adhesion and/or drug delivery, in addition to site-specific ligands such as lectins. Over the last 20 years a wide range of formulations has been developed for buccal drug delivery (tablet, patch, liquids and semisolids) but comparatively few have found their way onto the market. Currently, this route is restricted to the delivery of a limited number of small lipophilic molecules that readily cross the buccal mucosa. However, this route could become a significant means for the delivery of a range of active agents in the coming years, if the barriers to buccal drug delivery are overcome. In particular, patient acceptability and the successful systemic delivery of large molecules (proteins, oligonucleotides and polysaccharides) via this route remains both a significant opportunity and challenge, and new/improved technologies may be required to address these.

The basics and underlying mechanisms of mucoadhesion.

Mucoadhesion is where two surfaces, one of which is a mucous membrane, adhere to each other. This has been of interest in the pharmaceutical sciences in order to enhance localised drug delivery, or to deliver 'difficult' molecules (proteins and oligonucleotides) into the systemic circulation. Mucoadhesive materials are hydrophilic macromolecules containing numerous hydrogen bond forming groups, the carbomers and chitosans being two well-known examples. The mechanism by which mucoadhesion takes place has been said to have two stages, the contact (wetting) stage followed by the consolidation stage (the establishment of the adhesive interactions). The relative importance of each stage will depend on the individual application. For example, adsorption is a key stage if the dosage form cannot be applied directly to the mucosa of interest, while consolidation is important if the formulation is exposed to significant dislodging stresses. Adhesive joint failure will inevitably occur as a result of overhydration of a dosage form, or as a result of epithelia or mucus turnover. New mucoadhesive materials with optimal adhesive properties are now being developed, and these should enhance the potential applications of this technology.

Mucoadhesive, triclosan-loaded polymer microspheres for application to the oral cavity: preparation and controlled release characteristics.

The aim of this study was to develop mucoadhesive microspheres that can be utilised for the controlled release of triclosan in oral-care formulations, specifically dental pastes. Using a double-emulsion solvent evaporation technique, triclosan was incorporated into microspheres that were prepared from Gantreztrade mark MS-955, Carbopol 974P, polycarbophil or chitosan and the profiles for its release were established under simulated 'in use' conditions. Triclosan was rapidly released into a sodium lauryl sulphate containing buffer from all but the chitosan microspheres. The release of triclosan from microspheres suspended in a non-aqueous paste, was found to be sustained over considerable time-periods, which were influenced strongly by the nature of the polymeric carrier. For microspheres that were fabricated from Gantrez, Carbopol or polycarbophil, the release appeared to obey zero-order kinetics whereas in the case of chitosan-derived vehicles, the release profile fitted the Baker and Lonsdale model. The work has demonstrated that these polymeric microspheres, particularly those of chitosan, are promising candidates for the sustained release of triclosan in the oral cavity.

Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor.

The cytokine erythropoietin (Epo) is tissue-protective in preclinical models of ischemic, traumatic, toxic, and inflammatory injuries. We have recently characterized Epo derivatives that do not bind to the Epo receptor (EpoR) yet are tissue-protective. For example, carbamylated Epo (CEpo) does not stimulate erythropoiesis, yet it prevents tissue injury in a wide variety of in vivo and in vitro models. These observations suggest that another receptor is responsible for the tissue-protective actions of Epo. Notably, prior investigation suggests that EpoR physically interacts with the common beta receptor (betacR), the signal-transducing subunit shared by the granulocyte-macrophage colony stimulating factor, and the IL-3 and IL-5 receptors. However, because betacR knockout mice exhibit normal erythrocyte maturation, betacR is not required for erythropoiesis. We hypothesized that betacR in combination with the EpoR expressed by nonhematopoietic cells constitutes a tissue-protective receptor. In support of this hypothesis, membrane proteins prepared from rat brain, heart, liver, or kidney were greatly enriched in EpoR after passage over either Epo or CEpo columns but covalently bound in a complex with betacR. Further, antibodies against EpoR coimmunoprecipitated betacR from membranes prepared from neuronal-like P-19 cells that respond to Epo-induced tissue protection. Immunocytochemical studies of spinal cord neurons and cardiomyocytes protected by Epo demonstrated cellular colocalization of Epo betacR and EpoR. Finally, as predicted by the hypothesis, neither Epo nor CEpo was active in cardiomyocyte or spinal cord injury models performed in the betacR knockout mouse. These data support the concept that EpoR and betacR comprise a tissue-protective heteroreceptor.

In situ evaluation of drug-loaded microspheres on a mucosal surface under dynamic test conditions.

The ability of polymeric microspheres fabricated from Carbopol, polycarbophil, chitosan or Gantrez to retain a model hydrophilic drug (sodium fluorescein) was evaluated in situ, using a dynamic test system and image analysis. This technique used oesophageal tissues and simulated the physiological conditions within the oral cavity in terms of temperature, humidity and saliva flow. The point of sample application was observed over a 2h period by means of a digital camera. No significant differences in fluorescein colour intensity was obtained for the Gantrez and chitosan particles over 100min, indicate that these two polymers provide the possibility of prolonged action. Carbopol and polycarbophil particles became rapidly swollen and released the sodium fluorescein completely within 20min. It was concluded that the test system allowed the evaluation of the in situ behaviour of test particles, in terms of their ability to retain a water-soluble, coloured marker in 'dynamic' test conditions, and that chitosan and Gantrez were promising candidates for the production of mucoadhesive, sustained-release microspheres for water-soluble materials.