Donor Chamber Research Articles

Improving oral absorption of a rapidly crystallizing parent drug using prodrug strategy: Comparison of phosphate versus glycine based prodrugs

With an increasing number of Biopharmaceutical Classification System (BCS) II/IV pipeline compounds, solubilizing and supersaturating formulation strategies are becoming prevalent. Beyond formulation and solid form strategies, prodrugs are also employed to overcome solubility-limited absorption of poorly water-soluble compounds. Prodrugs can potentially yield supersaturated systems upon conversion to the parent drug intraluminally and thus enhance absorption. However, supersaturation also increases the driving force for crystallization, resulting in low solution concentrations, which can potentially negate the advantage of prodrugs. In this work, two unique solubility-enhancing prodrugs, phosphate and glycine esters, were investigated for a rapidly crystallizing parent drug. Ex vivo absorption studies using rat tissue and in vivo studies in dogs were performed. Conversion rate of the phosphate prodrug to the parent was dependent on the milieu and increased ∼24-fold in the presence of intestinal contents as medium and tissue relative to neat buffer. In contrast, conversion of the glycine prodrug was minimal under any conditions tested, suggesting that the conversion occurs after absorption into the enterocytes. Phosphate prodrug showed a non-linear increase in parent drug absorptive flux across rat intestinal tissue with concentration when intestinal contents were used as donor media. This was attributed to rapid conversion and high supersaturation of the parent drug which subsequently resulted in crystallization at high doses in the donor chamber. Glycine prodrug did not undergo complete conversion at high doses and was absorbed unchanged on the basolateral side, indicating saturation of the converting enzymes in the enterocytes. The combined flux (parent drug and glycine) showed a linear increase with dose and crystallization was not observed. Under physiological conditions, glycine prodrug that is absorbed unchanged from the intestine can potentially undergo complete conversion in hepatocytes after absorption and make the parent drug systemically available. Thus, glycine prodrug provided overall higher absorption compared to phosphate prodrug. The observed flux levels for both the prodrugs were higher compared to the parent drug alone, highlighting an advantage to use of a prodrug strategy to improve absorption of such compounds. Oral dosing in a dog PK study revealed that the bioavailability using the phosphate prodrug was ∼50% whereas, it was ∼100% with glycine prodrug, supporting the in vitro observations.

Benefits of combining supersaturating and solubilizing formulations – Is two better than one?

A variety of enabling formulations has been developed to address poor oral drug absorption caused by insufficient dissolution in the gastrointestinal tract. As the in vivo performance of these formulations is a result of a complex interplay between dissolution, digestion and permeation, development of suitable in vitro assays that captures these phenomena are called for. The enabling-absorption (ENA) device, consisting of a donor and receiver chamber separated by a semipermeable membrane, has successfully been used to study the performance of lipid-based formulations. In this work, the ENA device was prepared with two different setups (a Caco-2 cell monolayer and an artificial lipid membrane) to study the performance of a lipid-based formulation (LBF), an amorphous solid dispersion (ASD) and the potential benefit of combining the two formulation strategies. An in vivo pharmacokinetic study in rats was performed to evaluate the in vitro-in vivo correlation. In the ENA, high drug concentrations in the donor chamber did not translate to a high mass transfer, which was particularly evident for the ASD as compared to the LBF. The solubility of the polymer used in the ASD was strongly affected by pH-shifts in vitro, and the ph_dependence resulted in poor in vivo performance of the formulation. The dissolution was however increased in vitro when the ASD was combined with a blank lipid-based formulation. This beneficial effect was also observed in vivo, where the drug exposure of the ASD increased significantly when the ASD was co-administered with the blank LBF. To conclude, the in vitro model managed to capture solubility limitations and strategies to overcome these for one of the formulations studied. The correlation between the in vivo exposure of the drug exposure and AUC in the ENA was good for the non pH-sensitive formulations. The deconvoluted pharmacokinetic data indicated that the receiver chamber was a better predictor for the in vivo performance of the drug, however both chambers provided valuable insights to the observed outcome in vivo. This shows that the advanced in vitro setting used herein successfully could explain absorption differences of highly complex formulations.

Continuous flow membrane microextraction as a clean method for detecting codeine and papaverine in biological samples using HPLC-UV

In this study, continuous-flow membrane microextraction (CFMME) was connected online with a high-performance liquid chromatography-UV detector (HPLC-UV) for the pre-concentration and high clean-up of papaverine and codeine in biological samples. The extraction cell was designed with donor and acceptor chambers separated by a sheet membrane. By adjusting the pH of the donor phase (10 mL, pH 11), the analyte molecules were extracted into the supported liquid membrane (SLM) (15 μL of 1-octanol). The donor solution was circulated through the donor chamber using a peristaltic pump and magnetically agitated by a bar stirrer placed near the membrane, enhancing the diffusion and convection flow of the targeted drugs from the donor solution to the SLM. Subsequently, by adjusting the acceptor solution to an acidic pH of 2 (100 μL), the drugs in ionic form were reversely extracted into the acceptor phase. The procedure exhibited desirable relative standard deviation (RSD%) of less than 3.70%, linear ranges of 7-600 ng mL-1 for codeine and 2.0-600 ng mL-1 for papaverine, and limits of detection of 2.0 ng mL-1 for codeine and 0.6 ng mL-1 for papaverine. The design of the extraction cell significantly improved the performance for determining targeted drugs in complex matrices such as plasma and urine.

Development of an In Vitro Methodology to Assess the Bioequivalence of Orally Disintegrating Tablets Taken without Water.

To assess the probability of bioequivalence (BE) between orally disintegrating tablets (ODTs) taken without water and conventional tablets (CTs) taken with water, an in vitro biorelevant methodology was developed using the BE Checker, which reproduces fluid shifts in the gastrointestinal tract and drug permeation. In addition to the fluid shift from the stomach to the small intestine, the process of ODT disintegration in a small amount of fluid in the oral cavity and the difference in gastric emptying caused by differences in water intake were incorporated into the evaluation protocol. Assuming a longer time to maximum plasma concentration after oral administration of ODTs taken without water than for CTs taken with water due to a delay in gastric emptying, the fluid shift in the donor chamber of the BE Checker without water was set longer than that taken with water. In the case of naftopidil ODTs and CTs, the values of the f2 function, representing the similarity of the permeation profiles, were 50 or higher when the fluid shift in ODTs taken without water was set at 1.5 or 2 times longer than that of the CTs taken with water. The values of the f2 function in permeation profiles of pitavastatin and memantine ODTs were both 62 when the optimized experimental settings for naftopidil formulations were applied. This methodology can be useful in formulation studies for estimating the BE probability between ODTs and CTs.

Method for Testing of Drugs Belonging to Substrates and Inhibitors of the Transporter Protein BCRP on Caco-2 Cells

Introduction. Breast cancer resistance protein (BCRP) is an efflux membrane transporter that controls the pharmacokinetics of a large number of drugs. Its activity may change when taking some endo- and exogenous substances, thus making it a link in drug interactions.Aim. The aim of the study was to develop a method for testing of drugs for belonging to BCRP substrates and inhibitors in vitro.Materials and methods. The work was performed on Caco-2 cells overexpressing BCRP, the cultivation was performed in a transwell-system consisting of the apical and basolateral chambers. Cells were seeded at the bottom of the apical chamber, which is a semipermeable membrane. Primarily, the transport of BCRP substrates: methotrexate, mitoxantrone and quercetin was evaluated in the concentration range of 1, 5, 10, and 50 μM in the direction from the basal chamber to the apical one (Papp b-a) and in the opposite direction (Papp a-b). The ratio Papp b-a / Papp a-b more than «2» characterizes the participation of transporter proteins in the transcellular transport of substances. To confirm the participation of BCRP in their transport the experiment was carried out with the addition of a transporter inhibitor, reserpine, to the transport medium at a concentration of 50 μM. The concentration of substrates in the chambers was analyzed by HPLC-MS/MS.Results and their discussion. The addition of methotrexate (1 μM), mitoxantrone (1 μM), and quercetin (1–10 μM) to both the apical or basolateral chambers of the transwell-system, their content in the recipient chamber was not detected. When methotrexate concentration became 5 μM the Papp b-a / Papp a-b ratio was 3.38 ± 0.08, which indicates the involvement of transporters in its transfer. The addition of methotrexate to the donor chamber at concentrations of 10 and 50 μM, Papp b-a / Papp a-b decreased to values below «2». At mitoxantrone concentration of 5 μM Papp b-a / Papp a-b was 2.72 ± 0.16. An increase in the concentration to 10 μM led to an increase in Papp b-a / Papp a-b to 6.18 ± 0.08. With a substance content of 50 μM the indicator decreased but remained above the value «2». In the quercetin concentration of 50 microns, Papp b-a / Papp was below "2". Reserpine reduced Papp b-a / Papp a-b of methotrexate by 3.31 times (p = 0.0002), which indicates the elimination of asymmetry in the transport of the substance. At a mitoxantrone concentration of 10 microns, reserpine reduced its Papp b-a / Papp a-b by 3.36 times (p < 0.0001). The results indicate the participation of BCRP in the control of the transfer of both substances through the cellular monolayer.Conclusion. A method of testing drugs belonging to BCRP substrates and inhibitors using methotrexate (5 μM) and mitoxantrone (10 μM) as marker substrates and reserpine (50 μM) as inhibitor was developed and tested on Caco-2 cells.

Effect of alkalinity on the diffusion of solvent-fractionated lignin through cellulose membranes

Mass transport of liberated lignin fragments from pits and fiber walls into black liquor is considered a determining step in the delignification process. However, our current understanding of the diffusion of lignin through cellulose and the influential parameter on this process is very limited. A comprehensive and detailed study of lignin mass transport through cellulosic materials is, therefore, of great importance. In this study, diffusion cell methodology is implemented to systematically investigate the transport of fractionated kraft lignin molecules through model cellulose membranes. Pulping is a complex process and lignin is very heterogenous material therefore to perform a more detailed study on lignin diffusion, we included an additional solvent fractionation step. One of the benefits of this method is that the setup can be adjusted to various experimental conditions allowing the complex chemical reactions occurring during pulping, which would affect the mass transfer of lignin, to be avoided. Here, the effects of the alkalinity of the aqueous solution and molecular weight of the kraft lignin molecules on their diffusion were investigated. Additionally, NMR spectroscopy, size exclusion chromatography, and UV/Vis spectroscopy were used to characterize the starting material and the molecules that passed through the membrane. Lignin molecules detected in the acceptor chamber of the diffusion cells had lower molecular weights, indicating a size fractionation between the donor and acceptor chamber. UV/Vis showed higher concentrations of ionized conjugated kraft lignin molecules in the acceptor chamber, which is a sign of chemical fractionation. This study suggests that the diffusion of lignin through small cellulose pores can be enhanced by decreasing the average molecular weight of the diffusing kraft lignin molecules and increasing alkalinity.

Abstract A022: Limonene as a vehicle in topical delivery of carvedilol for skin cancer chemoprevention

Abstract Skin cancer is currently one of the most common cancers and is defined as an abnormal growth of the skin cells. The goal of this study is to develop a topical delivery system of carvedilol with limonene as the vehicle. Limonene is a major component found in the rinds of citrus fruits and other herbs. It is one of many natural compounds that have demonstrated inhibitory activity in different types of cancers such as breast, lung, and stomach cancer. Furthermore, limonene has shown to increase skin permeation of drugs including carvedilol. The soft agar colony formation assay was used to evaluate anchorage-independent growth of JB6 mouse epidermal cells. This assay tested concentrations of carvedilol (0.1, 1.0 and 10 μM); limonene (1.0, 10, 100 and 1000 μM); and a combination of carvedilol (1.0 μM) and limonene (1.0, 10 or 100 μM). Results demonstrated that carvedilol and limonene as single treatment inhibited colony formation. The combination of carvedilol and limonene showed higher colony inhibition than single treatment, although not statistically significant. Franz diffusion cell was used to test for permeation of the drug. The device has two chambers separated by a membrane, which in this case is rat skin. The drugs, in vehicles of 40% polyethylene glycol (PEG) 400 in PBS or 40% PEG400 in PBS plus 5% limonene were applied into the donor chamber while the samples are collected through the receptor chamber as the drug permeates through the skin. The samples were collected at 16 hours, 20 hours, and 24 hours. Results showed that limonene significantly increased permeation. Therefore, our data indicate that limonene was able to increase skin permeation and the chemopreventive activity of carvedilol. This new formulation will be further examined in animal models for its efficacy against UV-induced skin cancer. Citation Format: Janell Wang, Lawrence Chao, Steven Yeung, Ying Huang. Limonene as a vehicle in topical delivery of carvedilol for skin cancer chemoprevention [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr A022.

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.

Passive absorption across gastrointestinal tissues in vitro and postharvest distribution of loline alkaloid in lambs

Loline alkaloid has suggested antimicrobial and anthelmintic properties with very low mammalian toxicity. There are several known derivatives of loline; N-formyl loline (NFL), N-acetyl loline (NAL), N-acetyl norloline (NANL), N-methyl loline (NML), and loline base. However, these must reach the abomasum or intestine and then be absorbed to have any potential effect. Therefore, passive absorption in isolated gastrointestinal tissues and distribution in the tissue of lambs were determined. Experiment 1: Lamb (n = 6) isolated gastrointestinal tissues were removed and mounted in an Ussing chamber. Approximately, 1 034 µg/g of loline and 22.1 µg/mL of caffeine were added to the donor chamber to measure loline flux at 0, 0.5, 1, and 2 hours. Experiment 2: Two, 12-week-old lambs were dosed with 52.5 mg/kg BW loline twice and were slaughtered to determine the distribution in gastrointestinal, organs, and blood. Passive absorption was 5% in ileum, <2% in ruminal or abomasal tissues. Loline base and NFL were passively absorbed across all tissues, with NAL and NANL only crossing small intestine tissues. Surprisingly, no caffeine crossed any tissues. Loline base and small amounts of NFL were in blood plasma, and loline base was also found in liver and kidneys. Results indicate either the majority of loline is not passively absorbed or membrane integrity was affected as suggested by lack of caffeine absorption.

Mass Transport of Lignin in Confined Pores.

A crucial step in the chemical delignification of wood is the transport of lignin fragments into free liquor; this step is believed to be the rate-limiting step. This study has investigated the diffusion of kraft lignin molecules through model cellulose membranes of various pore sizes (1–200 nm) by diffusion cells, where the lignin molecules diffuse from donor to acceptor cells through a membrane, where diffusion rate increases by pore size. UV–vis spectra of the donor solutions showed greater absorbance at higher wavelengths (~450 nm), which was probably induced by scattering due to presence of large molecules/clusters, while acceptor samples passed through small pore membranes did not. The UV–vis spectra of acceptor solutions show a characteristic peak at around 350 nm, which corresponds to ionized conjugated molecules: indicating that a chemical fractionation has occurred. Size exclusion chromatography (SEC) showed a difference in the molecular weight (Mw) distribution between lignin from the donor and acceptor chambers. The results show that small pore sizes enable the diffusion of small individual molecules and hinder the transport of large lignin molecules or possible lignin clusters. This study provides more detail in understanding the mass transfer events of pulping processes.

The Interactions and Release Kinetics of Sodium Hyaluronate Implemented in Nonionic and Anionic Polymeric Hydrogels, Studied by Immunoenzymatic ELISA Test.

Hyaluronan is a natural polymer that was introduced to wound therapy. Formulations based on synthetic polymers such as methylcellulose (MC) and polyacrylic acid (PA) containing hyaluronan (HA) were proposed for the development of prospective wound-healing preparations. The formulations of different carrier concentrations containing a fixed amount of HA were prepared, and their viscosity was measured. The HA release was evaluated by employing the apparatus paddle over a disc. The hydrogels were introduced to the donor chamber, and HA was released to the pH = 7.4 buffer. The amount of HA released was obtained using the ELISA test. The release was analyzed on the basis of different kinetic models: zero-, first-, and second-order kinetics, as well as Higuchi and Korsmeyer–Peppas equations. The release rate constants and the half release time were calculated from these equations. According to the value of the coefficient of the determination, the best model describing the observed process was selected. The comparison between the dissolution profiles was carried out by calculating the difference factor f1 and the similarity factor f2. The interaction between the hydrogel components was investigated by Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) measurements. The study revealed that the zero-order equation best described the release of HA from the formulations studied. The FTIR research and the DSC study showed the intermolecular interaction between HA chains in MC-based compositions, as well as between HA and the synthetic polymer in the PA-based formulations. The study revealed that the formulation with a higher concentration of synthetic polymer may prolong the release of HA. The obtained results indicated that the proposed hydrogels have potential for wound healing and may accelerate skin regeneration.

P62 accumulates through neuroanatomical circuits in response to tauopathy propagation

In Alzheimer’s disease and related tauopathies, trans-synaptic transfer and accumulation of pathological tau from donor to recipient neurons is thought to contribute to disease progression, but the underlying mechanisms are poorly understood. Using complementary in vivo and in vitro models, we examined the relationship between these two processes and neuronal clearance. Accumulation of p62 (a marker of defective protein clearance) correlated with pathological tau accumulation in two mouse models of tauopathy spread; Entorhinal Cortex-tau (EC-Tau) mice where tau pathology progresses in time from EC to other brain regions, and PS19 mice injected with tau seeds. In both models and in several brain regions, p62 colocalized with human tau in a pathological conformation (MC1 antibody). In EC-Tau mice, p62 accumulated before overt tau pathology had developed and was associated with the presence of aggregation-competent tau seeds identified using a FRET-based assay. Furthermore, p62 accumulated in the cytoplasm of neurons in the dentate gyrus of EC-Tau mice prior to the appearance of MC1 positive tauopathy. However, MC1 positive tau was shown to be present at the synapse and to colocalize with p62 as shown by immuno electron microscopy. In vitro, p62 colocalized with tau inclusions in two primary cortical neuron models of tau pathology. In a three-chamber microfluidic device containing neurons overexpressing fluorescent tau, seeding of tau in the donor chamber led to tau pathology spread and p62 accumulation in both the donor and the recipient chamber. Overall, these data are in accordance with the hypothesis that the accumulation and trans-synaptic spread of pathological tau disrupts clearance mechanisms, preceding the appearance of obvious tau aggregation. A vicious cycle of tau accumulation and clearance deficit would be expected to feed-forward and exacerbate disease progression across neuronal circuits in human tauopathies.

Use of single particle ICP-MS to estimate silver nanoparticle penetration through baby porcine mucosa

Children are potentially exposed to products that contain nanoparticles (NPs). In particular, silver NPs are commonly present both in products used by and around children, primarily due to their antibacterial properties. However, very few data are available regarding the ability of silver NPs to penetrate through the oral mucosa in children. In the present work, we used baby porcine buccal mucosa mounted on vertical Franz diffusion cells, as an in vitro model to investigate penetration of silver NPs (19 ± 5 nm). Permeability experiments were performed using pristine physiologically-relevant saline solution in the receiver chamber and known concentrations of NPs or ions in the donor chamber; conditions mimicked the in vivo physiological pH conditions. After physicochemical characterization of silver nanoparticles dispersed in physiological solution, we evaluated the passage of ions and NPs through the mucosa, using single particle inductively coupled plasma mass spectrometry. A flux of 4.1 ± 1.7 ng cm−2 min−1 and a lag time of 159 ± 17 min were observed through mucosa exposed to silver nanoparticles. The latter suggests nanoparticle penetration through the baby porcine mucosa and release Ag+ ions in the receptor fluid, as confirmed by computational model. Due to physiological similarity between human and pig membranes it is reasonable to assume that a trans-oral mucosa penetration could occur in children upon contact with silver nanoparticles.

A novel versatile flow-donor chamber as biorelevant ex-vivo test assessing oral mucoadhesive formulations

Oral transmucosal drug delivery is a non-invasive administration route for rapid therapeutic onset and greater bioavailability avoiding the first-pass metabolism. Mucoadhesive formulations are advantageous as they may retain the drug at the administration site. Proper equipment to assess mucoadhesive properties and corresponding drug absorption is fundamental for the development of novel drug delivery systems. Here we developed a new flow-through donor chamber for well-established diffusion cells, and we tested the effects on drug and formulation retention in situ of adding mucoadhesive polymers or mesoporous silica particles to a reference formulation. Mesoporous silica particles are of particular interest as they may be used to encapsulate and retain drug molecules. Compared to other ex-vivo methods described in literature for assessing mucoadhesive performance and transmucosal drug delivery, this new donor chamber provides several advantages: i) it reflects physiological conditions better as a realistic saliva flow can be provided over the administration site, ii) it is versatile since it can be mounted on any kind of vertical diffusion cell allowing simultaneous detection of drug retention at the administration site and drug permeation through the tissue, and iii) it enables optical quantification of formulations residence time aided by image processing. This new flow-through donor diffusion cell set-up proved sensitive to differentiate a reference formulation from one where 20 %(w/w) Carbomer was added (to further improve the mucoadhesive properties), with respect to both drug and formulation residence times. We also found that mesoporous silica particles, investigated as particles only and mixed together with the reference formulation, gave very similar drug and formulation retention to what we observed with the mucoadhesive Carbomer. However, after some time (>30 min) it became obvious that the tablet excipients in the reference formulation promote particle retention on the mucosa. This work provides a new simple and versatile biorelevant test for the evaluation of oral mucoadhesive formulations and paves the way for further studies on mesoporous silica particles as valuable excipients for enhancing oral mucoadhesion.

Bioequivalence of Oral Drug Products in the Healthy and Special Populations: Assessment and Prediction Using a Newly Developed In Vitro System "BE Checker".

In order to assess and predict the bioequivalence (BE) of oral drug products, a new in vitro system “BE checker” was developed, which reproduced the environmental changes in the gastrointestinal (GI) tract by changing the pH, composition, and volume of the medium in a single chamber. The dissolution and membrane permeation profiles of drugs from marketed products were observed in the BE checker under various conditions reflecting the inter-patient variations of the GI physiology. As variable factors, initial gastric pH, gastric emptying time, and GI agitation strength were varied in vitro. Dipyridamole, a basic drug, showed rapid and supersaturated dissolution when the paddle speed in the donor chamber was 200 rpm, which corresponds to the high agitation strength in the stomach. In contrast, supersaturated dissolution disappeared, and the permeated amount decreased under the conditions with a slow paddle speed (100 and 50 rpm) and short gastric emptying time (10 min). In those conditions, disintegration of the formulation was delayed, and the subsequent dissolution of dipyridamole was not completed before the fluid pH was changed to neutral. Similar results were obtained when the initial gastric pH was increased to 3.0, 5.0, and 6.5. To investigate that those factors also affect the BE of oral drug products, dissolution and permeation of naftopidil from its ordinary and orally disintegrating (OD) tablets were observed in the BE checker. Both products showed the similar dissolution profiles when the paddle speed and gastric emptying time were set to 100 rpm and 10 or 20 min, respectively. However, at a low paddle speed (50 rpm), the dissolution of naftopidil from ordinary tablets was slower than that from the OD tablets, and the permeation profiles became dissimilar. These results indicated the possibility of the bioinequivalence of some oral formulations in special patients whose GI physiologies are different from those in the healthy subjects. The BE checker can be a highly capable in vitro tool to assess the BE of oral drug products in various populations.

Dose-Dependent Solubility-Permeability Interplay for Poorly Soluble Drugs under Non-Sink Conditions.

We investigated the solubility–permeability interplay using a solubilizer additive under non-sink conditions. Sodium lauryl sulfate (SLS) was used as a solubilizer additive. The solubility and permeability of two poorly soluble drugs at various doses, with or without SLS, were evaluated by flux measurements. The total permeated amount of griseofulvin, which has high permeability, increased by the addition of SLS. On the other hand, triamcinolone, which has low permeability, showed an almost constant rate of permeation regardless of the SLS addition. The total permeated amount of griseofulvin increased by about 20–30% when the dose amount exceeded its solubility, whereas its concentration in the donor chamber remained almost constant. However, the total permeated amount of triamcinolone was almost constant regardless of dose amount. These results suggest that the permeability of the unstirred water layer (UWL) may be affected by SLS and solid drugs for high-permeable drugs. The effect of solid drugs could be explained by a reduction in the apparent UWL thickness. For the appropriate evaluation of absorption, it would be essential to consider these effects.

Evaluation of Electroosmotic Flow Promoted By a Porous Microneedle Array

A transdermal drug delivery (TDD) system has been an attractive method to delivery needed drug into our bodies directly with few side effect, and electroosmotic flow (EOF) is known as the method to enhance TDD and even the extraction of interstitial fluid (ISF) which contains biomarkers such as glucose. The limitation of these transdermal penetration / extraction arises from stratum corneum (SC), an outermost layer of skin. SC is highly resistive and functions as a barrier for harmful molecules, which makes it difficult to apply direct currents to the skin by mild and safe voltage and to deliver larger molecular drugs (>600 Da) into the skin1. The microneedle array (MNA) is one of the promising approaches to break through SC without pain, and solve problems of the electrical and the drug pass. Recently, we have realized organic polymer-based porous microneedles (PMN) made of poly(glycidyl methacrylate) (PGMA) which were prepared by the combination of molding process and the porogen method 2. In this study, we developed a PMN inducing the EOF for efficient TDD and extraction of ISF (Figure 1a). Furthermore, in order to enhance efficiency in the induction of the EOF, negatively-charged hydrogel was embedded in the porous channels of the PMN.The modified PMN was prepared by the following steps. Briefly, naked PMN was prepared by the step described in the report2. After that, the naked PMN chips were immersed in an aqueous solution containing 0.05 M of AMPS, 0.10 M of MBAAm, ammonium persulfate and tetramethylenediamine at 4 ℃ for more than 8 h, followed by thermal copolymerization at 70 ℃ for more than 8 h.First, we evaluated the property of water transport by EOF. A chip of PAMPS-modified PGMA was bound with side-by-side Franz cells with a horizontal capillary, and constant DC currents were applied by a source meter. McIlvaine buffer (pH 7.0) was used as the electrolyte. Flow velocity was calculated from the moving of the water surface in the capillary. The flow velocity of the EOF increased by the modification with AMPS (Figure 1b). The reason was that the negative charge was increased by the AMPS modification and that promoted the EOF.In order to evaluate the property of large molecule transport, the same system as water transport was employed as well. The Franz cells were poured with McIlvaine buffer (pH 6.0) and 0.75 mg/mL FITC-dextran (10,000 Da, pKa 6.4) was added to the donor chamber. We applied a constant DC current in direction from the donor side to the receptor side, and a 100 µL portion was collected from the receptor chamber every hour for analysis by a fluorescence spectrophotometer. From the result, we found that PAMPS significantly contributes to transport for the large molecule transport (Figure 1c). This might be due to dense polymer chains of PMAPS capturing large molecules such as dextran.Finally, the transdermal injection was demonstrated by enzymatic fructose / O2 battery into pig’s skin (Figure 1d). The injection of FITC-dextran at an anode was evaluated the PMN modified with 0.05 M AMPS and the cotton with 0.3 mL McIlvaine buffer (pH 6.0) containing 0.2 M D-fructose and 0.75 mg/mL FITC-dextran. The cross-sectional fluorescence image of the skin was observed after the application of the current around 0.2 mA/cm2 for 1 hour. These results showed that this novel PMN with negatively charged hydrogel can be a desirable tool for enhancing the injection of large molecules such as drugs and nutrients.Reference[1] Brown, M. B, Martin, G. P., Jones, S. A. & Akomeah, F. K. Dermal and transdermal drug delivery system: Current and future prospects. Drug Deliv. J. Deliv. Target. Ther. Agents 13, 175-187 (2006).[2] Liu, L., Kai, H., Nagamine, K., Ogawa, Y. & Nishizawa, M. Porous polymer microneedles with interconnecting microchannels for rapid fluid transport. RSC Adv. 6, 48630-48635 (2016). Figure 1

FORMULATION AND EVALUATION OF CELECOXIB CREAM AND ITS RELEASED STUDY

Objective: The purpose of this study was to formulate and evaluate of Celecoxib cream and it’s in vitro release study.&#x0D; Methods: The release study was conducted, using dialysis cellulose membrane, in Franz cells. The donor chamber was filled with phosphate buffer pH 7.4, released medium were analyzed by UV-Vis spectrophotometer at 250 nm. Kinetics model was used for calculations. The cream was followed by different evaluations like pH measurement, homogeneity, spreadability, stability study, drug content, SEM, XRD studies and skin irritation test was used for the reliability of physical conditions and chemical relation. DD solver and SPSS were used for statistical analyzation of the data.&#x0D; Results: The best in vitro drug release profile achieved with thyme oil in Celecoxib cream. Formulation F2 showed the highest (83%) released. The results of the Celecoxib (1%) were suitable in all constraints. The prepared Celecoxib cream was encouraging for the formulation of transdermal drug delivery.&#x0D; Conclusion: The Celecoxib cream was successfully prepared and could be beneficial for transdermal drug delivery.

Gelation of the internal core of liposomes as a strategy for stabilization and modified drug delivery II. Theoretical analysis and modelling of in-vitro release experiments.

PEG-DMA was incorporated in unilamellar liposomes. PEG-DMA crosslinking by photo-induced radical reaction transforms the liquid aqueous core of the liposome into a hydrogel. The molecular weight of PEG-DMA significantly influences both structural and release properties of these hybrid nanosystems, by affecting both membrane permeability and diffusional properties of the inner core. Release studies of 5-(6) carboxyfluorescein from Conventional Liposomes (CL) and Gel-in-Liposome (GiL) systems were carried out in a vertical Franz Diffusion Cell. A detailed transport model is proposed, aimed at describing the entire drug diffusive pathway from the vesicles' inner core, through the double-layer membrane, into the buffer solution in the donor chamber of the Franz Cell and from there to the receptor chamber, where withdrawals are performed to evaluate the released drug concentration. The model permits to give a quantitative estimate of the diffusional resistances offered by the inner core (liquid or gelled) and by the double-layer membrane for CLs and different GiLs systems. The theoretical analysis of experimental release data strongly supports the basic assumption that, by varying the molecular weight of PEG-DMA, a different arrangement of the polymer within the liposomal structure and a different interaction with the membrane occur. PEG750-DMA decreases the transport resistance of the double layer membrane with respect to CLs, while PEG4000-DMA plays the opposite role. After gelation of the internal core, the diffusional resistance to drug transport inside GiLs becomes controlling, thus significantly slowing down drug release from these systems. Therefore, the combination of PEG-DMA with phospholipid vesicles appears an interesting strategy to develop sustained drug delivery systems.

Towards direct analysis of solid and liquid samples exploiting a 3D printed dialysis unit and sequential injection: Application for automated derivatization and determination of gamma-aminobutyric acid in foodstuff and beverages

In this work, a new design of a dialysis unit for direct analysis of solid and liquid samples is presented. The homemade unit was constructed using a 3D printer due to its simple and fast fabrication ability. The dialysis unit is composed of cylindrical-shaped donor and acceptor chambers. A stainless steel sieve is installed inside the donor chamber. SEM images clearly showed that the sieve prevented membrane blockage by suspension particles in the sample. Multiple dialysis units were connected to a sequential injection (SI) system for serial determination of gamma-aminobutyric acid (GABA) in solid and liquid samples. The dialysate from each dialysis unit was consecutively aspirated into the SI flow line for on-line derivatization of GABA with 2-hydroxy-1-naphthaldehyde (3.0% w/v). The derivative was detected spectrophotometrically at 425 nm. The linear calibration range extended to 1000 mg L−1 GABA (r2 > 0.99) with high precision (1.2 %RSD). The developed system was applied to analysis of dietary supplements, grains of germinated brown rice and milk. The samples were directly introduced into the donor chamber either as powder or liquids. The measured GABA content using the developed method was compared using high performance liquid chromatography, with good agreement using Pearson’s correlation (r2 = 0.9999). The method has high accuracy based on recovery studies (99.8 ± 1.5%) and high sample throughput (64 samples h−1).