International Journal of Pharmaceutics (v.499, #1-2)

Impregnation of Fenofibrate on mesoporous silica using supercritical carbon dioxide by Abir Bouledjouidja; Yasmine Masmoudi; Michiel Van Speybroeck; Laurent Schueller; Elisabeth Badens (1-9).
Display OmittedLow oral bioavailability can be circumvented by the formulation of the poorly water soluble drug in ordered mesoporous silica (OMS-L-7). Fenofibrate is an orally administered, poorly water-soluble active pharmaceutical ingredient (API), used clinically to lower lipid levels. Fenofibrate was loaded into silica using two methods: incipient wetness and supercritical impregnation. This study investigates the impact of loading and the impact of varying supercritical carbon dioxide (scCO2) processing conditions. The objective is to enhance Fenofibrate loading into silica while reducing degree of the drug crystallinity, so as to increase the drug's dissolution rate and its bioavailability. The comparison of both impregnation processes was made in terms of impregnation yields and duration as well as physical characterization of the drug.While incipient wetness method led to a Fenofibrate loading up to 300 mgdrug/gsilica in 48 h of impregnation, the supercritical impregnation method yielded loading up to 485 mgdrug/gsilica in 120 min of impregnation duration, at 16 MPa and 308 K, with a low degree of crystallinity (about 1%) comparable to the crystallinity observed via the solvent method. In addition to the enhancement of impregnation efficiency, the supercritical route provides a solvent-free alternative for impregnation.
Keywords: Supercritical fluids; Soaking in liquid; Ordered mesoporous silica (OMS-L-7); Fenofibrate; Drug delivery system;

Display OmittedSolid lipid nanoparticles (SLNs) conjugated with tamoxifen (TX) and lactoferrin (Lf) were applied to carry anticancer carmustine (BCNU) across the blood–brain barrier (BBB) for enhanced antiproliferation against glioblastoma multiforme (GBM). BCNU-loaded SLNs with modified TX and Lf (TX-Lf-BCNU-SLNs) were used to penetrate a monolayer of human brain-microvascular endothelial cells (HBMECs) and human astrocytes and to target malignant U87MG cells. The surface TX and Lf on TX-Lf-BCNU-SLNs improved the characteristics of sustained release for BCNU. When compared with BCNU-loaded SLNs, TX-Lf-BCNU-SLNs increased the BBB permeability coefficient for BCNU about ten times. In addition, TX-BCNU-SLNs considerably promoted the fluorescent intensity of intracellular acetomethoxy derivative of calcein (calcein-AM) in HBMECs via endocytosis. However, the conjugated Lf could only slightly increase the fluorescence of calcein-AM. Moreover, the order of formulation in the inhibition to U87MG cells was TX-Lf-BCNU-SLNs > TX-BCNU-SLNs > Lf-BCNU-SLNs > BCNU-SLNs. TX-Lf-BCNU-SLNs can be effective in infiltrating the BBB and delivering BCNU to GBM for future chemotherapy application.
Keywords: Tamoxifen; Lactoferrin; Blood–brain barrier; Glioblastoma multiforme; Solid lipid nanoparticle; Carmustine;

Impact of formulation and process variables on solid-state stability of theophylline in controlled release formulations by Maxwell Korang-Yeboah; Ziyaur Rahman; Dhaval Shah; Adil Mohammad; Suyang Wu; Akhtar Siddiqui; Mansoor A. Khan (20-28).
Display OmittedUnderstanding the impact of pharmaceutical processing, formulation excipients and their interactions on the solid-state transitions of pharmaceutical solids during use and in storage is critical in ensuring consistent product performance. This study reports the effect of polymer viscosity, diluent type, granulation and granulating fluid (water and isopropanol) on the pseudopolymorphic transition of theophylline anhydrous (THA) in controlled release formulations as well as the implications of this transition on critical quality attributes of the tablets. Accordingly, 12 formulations were prepared using a full factorial screening design and monitored over a 3 month period at 40 °C and 75%. Physicochemical characterization revealed a drastic drop in tablet hardness accompanied by a very significant increase in moisture content and swelling of all formulations. Spectroscopic analysis (ssNMR, Raman, NIR and PXRD) indicated conversion of THA to theophylline monohydrate (TMO) in all formulations prepared by aqueous wet granulation in as early as two weeks. Although all freshly prepared formulations contained THA, the hydration–dehydration process induced during aqueous wet granulation hastened the pseudopolymorphic conversion of theophylline during storage through a cascade of events. On the other hand, no solid state transformation was observed in directly compressed formulations and formulations in which isopropanol was employed as a granulating fluid even after the twelve weeks study period. The transition of THA to TMO resulted in a decrease in dissolution while an increase in dissolution was observed in directly compressed and IPA granulated formulation. Consequently, the impact of pseudopolymorphic transition of theophylline on dissolution in controlled release formulations may be the net result of two opposing factors: swelling and softening of the tablets which tend to favor an increase in drug dissolution and hydration of theophylline which decreases the drug dissolution.
Keywords: Solid-state stability; Dissolution; Pseudopolymorphic transition; Theophylline;

Display OmittedCompaction of multiple-unit pellet system (MUPS) tablets has been extensively studied in the past few decades but with marginal success. This study aims to investigate the formulation and process strategies for minimizing pellet coat damage caused by compaction and elucidate the mechanism of damage sustained during the preparation of MUPS tablets in a rotary tablet press. Blends containing ethylcellulose-coated pellets and cushioning agent (spray dried aggregates of micronized lactose and mannitol), were compacted into MUPS tablets in a rotary tablet press. The effects of compaction pressure and dwell time on the physicomechanical properties of resultant MUPS tablets and extent of pellet coat damage were systematically examined. The coated pellets from various locations at the axial and radial peripheral surfaces and core of the MUPS tablets were excavated and assessed for their coat damage individually. Interestingly, for a MUPS tablet formulation which consolidates by plastic deformation, the tablet mechanical strength could be enhanced without exacerbating pellet coat damage by extending the dwell time in the compaction cycle during rotary tableting. However, the increase in compaction pressure led to faster drug release rate. The location of the coated pellets in the MUPS tablet also contributed to the extent of their coat damage, possibly due to uneven force distribution within the compact. To ensure viability of pellet coat integrity, the formation of a continuous percolating network of cushioning agent is critical and the applied compaction pressure should be less than the pellet crushing strength.
Keywords: Coated pellets; Ethylcellulose coat; Rotary tableting; Dwell time; Compaction pressure; Pellet volume fraction;

Display OmittedInhaled nano-antibiotics have recently emerged as the promising bronchiectasis treatment attributed to the higher and more localized antibiotic exposure generated compared to native antibiotics. Antibiotic nanoparticle complex (or nanoplex in short) prepared by self-assembly complexation with polysaccharides addresses the major drawbacks of existing nano-antibiotics by virtue of its high payload and cost-effective preparation. Herein we developed carrier-free dry powder inhaler (DPI) formulations of ciprofloxacin nanoplex by spray drying (SD) and spray freeze drying (SFD). d-Mannitol and l-leucine were used as the drying adjuvant and aerosol dispersion enhancer, respectively. The DPI formulations were evaluated in vitro in terms of the (1) aerosolization efficiency, (2) aqueous reconstitution, (3) antibiotic release, and (4) antimicrobial activity against respiratory pathogen Pseudomonas aeruginosa. The SFD powders exhibited superior aerosolization efficiency to their SD counterparts in terms of emitted dose (92% versus 66%), fine particle fraction (29% versus 23%), and mass median aerodynamic diameter (3 μm versus 6 μm). The superior aerosolization efficiency of the SFD powders was attributed to their large and porous morphology and higher l-leucine content. While the SFD powders exhibited poorer aqueous reconstitution that might jeopardize their mucus penetrating ability, their antibiotic release profile and antimicrobial activity were not adversely affected.
Keywords: Pulmonary drug delivery; Antibiotic nanoparticles; Dry powder inhaler; Chronic lung infection; Cystic fibrosis;

Microstructural investigation using synchrotron radiation X-ray microtomography reveals taste-masking mechanism of acetaminophen microspheres by Zhen Guo; Xianzhen Yin; Congbiao Liu; Li Wu; Weifeng Zhu; Qun Shao; Peter York; Laurence Patterson; Jiwen Zhang (47-57).
Display OmittedThe structure of solid drug delivery systems has considerable influence on drug release behaviors from particles and granules and also impacts other properties relevant to release characteristics such as taste. In this study, lipid-based microspheres of acetaminophen were prepared to mask the undesirable taste of drug and therefore to identify the optimal formulation for drug release. Synchrotron radiation X-ray computed microtomography (SR-μCT) was used to investigate the fine structural architectures of microspheres non-destructively at different sampling times during drug release test, which were simultaneously determined to quantitatively correlate the structural data with drug release behaviors. The results demonstrated that the polymeric formulation component, namely, cationic polymethacrylate (Eudragit E100), was the key factor to mask the bitter taste of acetaminophen by inhibiting immediate drug release thereby reducing the interaction intensity of the bitter material with the oral cavity taste buds. The structure and morphology of the microspheres were found to be influenced by the shape and particle size of the drug, which was also an important factor for taste-masking performance. The quantitative analysis generated detailed structural information which was correlated well with drug release behaviors. Thus, SR-μCT has been proved as a powerful tool to investigate the fine microstructure of particles and provides a new approach in the design of particles for taste masking.
Keywords: Taste masking; Lipid microspheres; Dissolution test; Microstructure; Synchrotron radiation X-ray computed microtomography; Three dimensional reconstruction;

Display OmittedIn general, the intestinal absorption of small hydrophilic molecules and macromolecules like peptides, after oral administration is very poor. Absorption enhancers are considered to be one of the most promising agents to enhance the intestinal absorption of drugs. In this research, we focused on a gemini surfactant, a new type of absorption enhancer. The intestinal absorption of drugs, with or without sodium dilauramidoglutamide lysine (SLG-30), a gemini surfactant, was examined by an in situ closed-loop method in rats. The intestinal absorption of 5(6)-carboxyfluorescein (CF) and fluorescein isothiocyanate-dextrans (FDs) was significantly enhanced in the presence of SLG-30, such effect being reversible. Furthermore, the calcium levels in the plasma significantly decreased when calcitonin was co-administered with SLG-30, suggestive of the increased intestinal absorption of calcitonin. In addition, no significant increase in the of lactate dehydrogenase (LDH) activity or in protein release from the intestinal epithelium was observed in the presence of SLG-30, suggestive of the safety of this compound. These findings indicate that SLG-30 is an effective absorption-enhancer for improving the intestinal absorption of poorly absorbed drugs, without causing serious damage to the intestinal epithelium.
Keywords: Intestinal absorption; Absorption enhancer; Gemini surfactant; Poorly absorbed drug; Peptide drug; Calcitonin;

Transformation of an active pharmaceutical ingredient upon high-energy milling: A process-induced disorder in Biclotymol by Benjamin Schammé; Nicolas Couvrat; Pascal Malpeli; Emeline Dudognon; Laurent Delbreilh; Valérie Dupray; Éric Dargent; Gérard Coquerel (67-73).
Display OmittedThis study investigates for the first time the thermodynamic changes of Biclotymol upon high-energy milling at various levels of temperature above and below its glass transition temperature (T g). Investigations have been carried out by temperature modulated differential scanning calorimetry (TM-DSC) and X-ray powder diffraction (XRPD). Results indicate that Biclotymol undergoes a solid-state amorphization upon milling at T g  – 45 °C. It is shown that recrystallization of amorphous milled Biclotymol occurs below the glass transition temperature of Biclotymol (T g  = 20 °C). This displays molecular mobility differences between milled Biclotymol and quenched liquid. A systematic study at several milling temperatures is performed and the implication of T g in the solid-state transformations generally observed upon milling is discussed. Influence of analysis temperature with respect to interpretation of results was investigated. Finally, it is shown that co-milling Biclotymol with only 20 wt% of amorphous PVP allows a stable amorphous dispersion during at least 5 months of storage.
Keywords: Amorphous; Biclotymol; Glass transition; Milling; Pharmaceuticals; Polyvinylpyrrolidone; Stability;

Self-emulsifying excipient platform for improving technological properties of alginate–hydroxypropylcellulose pellets by Paolo Mannina; Lorena Segale; Lorella Giovannelli; Andrea Foglio Bonda; Franco Pattarino (74-80).
Display OmittedIn this work, alginate, alginate–pectin and alginate–hydroxypropylcellulose pellets were produced by ionotropic gelation and characterized. Ibuprofen was selected as model drug; it was suspended in the polymeric solution in crystalline form or dissolved in a self-emulsifying phase and then dispersed into the polymeric solution. The self-emulsifying excipient platform composed of Labrasol (PEG-8 caprylic/capric glycerides) and d-α-tocopherol polyethylene glycol 1000 succinate (TPGS), able to solubilize the drug was used to improve the technological and biopharmaceutical properties of the alginate pellets. The pellets had diameters between 1317 and 2026 μm and a high drug content (>51%). DSC analysis showed the amorphous state of drug in the pellets containing the self-emulsifying phase. All the systems restricted drug release in conditions simulating the gastric environment and made the drug completely available at a pH value typical for the intestine. Only alginate-HPC systems containing the drug solubilized into the self-emulsifying phase showed the ability to partially control the release of ibuprofen at neutral pH. The self-emulsifying excipient platform is a useful tool to improve technological and biopharmaceutical properties of alginate–HPC pellets.
Keywords: Alginates; Hydroxypropylcellulose; Pectin; Pellets; Modified release; Self-emulsifying;

Nanoparticle transport across in vitro olfactory cell monolayers by Oihane Gartziandia; Susana Patricia Egusquiaguirre; John Bianco; José Luis Pedraz; Manoli Igartua; Rosa Maria Hernandez; Véronique Préat; Ana Beloqui (81-89).
Display OmittedDrug access to the CNS is hindered by the presence of the blood–brain barrier (BBB), and the intranasal route has risen as a non-invasive route to transport drugs directly from nose-to-brain avoiding the BBB. In addition, nanoparticles (NPs) have been described as efficient shuttles for direct nose-to-brain delivery of drugs. Nevertheless, there are few studies describing NP nose-to-brain transport. Thus, the aim of this work was (i) to develop, characterize and validate in vitro olfactory cell monolayers and (ii) to study the transport of polymeric- and lipid-based NPs across these monolayers in order to estimate NP access into the brain using cell penetrating peptide (CPPs) moieties: Tat and Penetratin (Pen). All tested poly(d,l-lactide-co-glycolide) (PLGA) and nanostructured lipid carrier (NLC) formulations were stable in transport buffer and biocompatible with the olfactory mucosa cells. Nevertheless, 0.7% of PLGA NPs was able to cross the olfactory cell monolayers, whereas 8% and 22% of NLC and chitosan-coated NLC (CS-NLC) were transported across them, respectively. Moreover, the incorporation of CPPs to NLC surface significantly increased their transport, reaching 46% of transported NPs. We conclude that CPP-CS-NLC represent a promising brain shuttle via nose-to-brain for drug delivery.
Keywords: Nose-to-brain delivery; NLC; Nanoparticles; Lipid nanoparticles; CPP; Olfactory mucosa;

Display OmittedAmorphous solid dispersions have for many years been a focus in oral formulations, especially in combination with a hot-melt extrusion process. The present work targets a novel approach with a system based on a fatty acid, a polymer and an inorganic carrier. It was intended to adsorb the acidic lipid by specific molecular interactions onto the solid carrier to design disorder in the alkyl chains of the lipid. Such designed lipid microdomains (DLM) were created as a new microstructure to accommodate a compound in a solid dispersion. Vibrational spectroscopy, X-ray powder diffraction, atomic force microscopy as well as electron microscopic imaging were employed to study a system of stearic acid, hydroxypropylcellulose and aluminum magnesium silicate. β-carotene was used as a poorly water-soluble model substance that is difficult to formulate with conventional solid dispersion formulations. The results indicated that the targeted molecular excipient interactions indeed led to DLMs for specific compositions. The different methods provided complementary aspects and important insights into the created microstructure. The novel delivery system appeared to be especially promising for the formulation of oral compounds that exhibit both high crystal energy and lipophilicity.
Keywords: Hot-melt extrusion; Acidic lipid; Inorganic carrier; Designed lipid microdomains; Interactions;

Formulation and in vitro efficacy of liposomes containing the Hsp90 inhibitor 6BrCaQ in prostate cancer cells by Félix Sauvage; Silvia Franzè; Alexandre Bruneau; Mouad Alami; Stéphanie Denis; Valérie Nicolas; Sylviane Lesieur; François-Xavier Legrand; Gillian Barratt; Samir Messaoudi; Juliette Vergnaud-Gauduchon (101-109).
Display Omitted6BrCaQ is a promising anti-cancer agent derived from novobiocin, which has been shown to inhibit Hsp90. 6BrCaQ was loaded into nanometer-scaled phospholipid vesicles (liposomes) suitable for drug delivery to solid tumors. The effective incorporation of the drug within the phospholipid bilayer was investigated by differential scanning calorimetry. Liposomal 6BrCaQ showed good activity on PC-3 cell lines in vitro in terms of apoptosis induction and cell growth arrest in G2/M. Liposomes containing 6BrCaQ were also shown to slow down migration of PC-3 cells in presence of chemokine ligand 2 and to synergize with doxorubicin. Several Hsp90 targeting molecules like geldanamycin induce accumulation of Hsp70, leading to cytoprotection and often correlated with poor prognosis. In this study, we did not report any Hsp70 induction after treatment with liposomal 6BrCaQ but a decrease in Hsp90 and CDK-4 protein expression, indicating an effect on the chaperon machinery. Liposomal encapsulation of 6BrCaQ revealed promising anti-cancer effects and a better understanding of its mechanism of action.
Keywords: Novobiocin analog; Hsp90; Liposomes; Apoptosis; Cell cycle; Migration;

Fabrication of pluronic and methylcellulose for etidronate delivery and their application for osteogenesis by Aparna Sai Laxmi Rangabhatla; Vimon Tantishaiyakul; Kwunchit Oungbho; Onpreeya Boonrat (110-118).
Display OmittedNovel hydrogels were prepared by blending 4% (w/w) methylcellulose (MC) with various concentrations of 12, 14, 16, 18 and 20% (w/w) pluronic F127 (PF) to form injectable implant drug delivery systems. The blends formed gels using lower concentrations of PF compared to when using PF alone. Etidronate sodium (EDS) at a concentration of 4 × 10−3  M was loaded into these blends for producing an osteogenesis effect. The pure gels or EDS loaded gels exhibited cytocompatibility to both the osteoblast (MC3T3-E1) and myoblast (C2C12) cell lines whereas the gels of 16PF, 18PF and 20PF were very cytotoxic to the cells. The EDS loaded gels demonstrated significantly greater alkaline phosphatase (ALP) activities compared to the pure gels. The longer exposure time periods of the samples to the cells, the greater was the ALP activity. These EDS loaded gels significantly increased proliferation of both cell lines thus indicating a bone regeneration effect. The PF/MC blends prolonged the in vitro release of EDS for more than 28 days. Based on the in vitro degradation test, the MC extensively improved the gel strength of the PF and delayed the degradation of the gels thus making them more functional for a sustained drug delivery for osteogenesis.
Keywords: Methylcellulose; Pluronic F127; Etidronate sodium; Alkaline phosphatase activity; MC3T3-E1; C2C12;

Communication from the periphery to the hypothalamus through the blood–brain barrier: An in vitro platform by João Pedro Martins; Cecília Juliana Alves; Estrela Neto; Meriem Lamghari (119-130).
Display OmittedOne of the major routes of communication from the peripheral systems to the hypothalamus, the core structure of body homeostasis, is the humoral transmission through the blood–brain barrier (BBB). The BBB cultures are the in vitro model of choice to depict the mechanisms behind blood–brain interplay. Still, this strategy excludes the integration of the brain tissue response and, therefore, the resulting output might be limited. In this study, two in vitro assays were established: BBB coculture model and hypothalamic organotypic cultures. The combination of these two assays was used as a platform to address the two critical steps in the humoral transmission through the BBB to the brain: blood-BBB/BBB-brain.The in vitro model of the BBB was performed according to a coculture system using a brain microvascular endothelial cell line (bEnd.3) and primary astrocytes. The expression of junctional molecules as claudin-5, ZO-1, occludin and VE-cadherin was observed in the bEnd.3 cell–cell contact, confirming the BBB phenotype of these endothelial cells. Moreover, the transendothelial electrical resistance (TEER) values (71.1 ± 9.4 Ω ×  cm2) and the permeability coefficients (Pe) obtained in the transendothelial flux test (3.3 ± 0.11 × 10−6  cm/sec) support high integrity of the established barrier. The hypothalamic organotypic cultures were prepared from 8-days-old C57Bl/6 mice brains, based on the air-medium interface culture method. High cell viability (82 ± 9.6%) and a dense neuronal network were achieved. The stimulation with dexamethasone resulted in an increased neuropeptide (NPY) expression, confirming the responsiveness of the neuronal system of these organotypic cultures.After optimization and characterization of each assay, the functionality of the platform was validated through the evaluation of the hypothalamic response to deep wound encompassing skin and muscle in mice. Results allowed to identify increased NPY activity in hypothalamic slices in response to peripheral signals within the plasma from wounded animals when compared with non-injured animals after surpassing and/or interacting with the BBB. This differential NPY response between the different animal conditions validated the functionality of the in vitro platform. In conclusion, this approach can be greatly anticipated as a useful tool for studying biologic or pharmacological circulating molecules and their impact on the hypothalamic activity.
Keywords: In vitro tools; Hypothalamus; Periphery-to-brain communication; Blood–brain barrier; Organotypic cultures;

A novel, anisamide-targeted cyclodextrin nanoformulation for siRNA delivery to prostate cancer cells expressing the sigma-1 receptor by Kathleen A. Fitzgerald; Meenakshi Malhotra; Matt Gooding; Florence Sallas; James C. Evans; Raphael Darcy; Caitriona M. O’Driscoll (131-145).
Display OmittedProstate cancer is a leading cause of cancer-related death in men and RNA interference (RNAi) has emerged as a potential therapeutic option. However, the absence of a safe and specific delivery vector remains a major obstacle to the clinical application of RNAi. Cyclodextrin derivatives are known to be efficient delivery systems with low toxicity in a variety of cell types. In this study, a cationic cyclodextrin derivative was synthesized to complex siRNA. The nanoparticle was then further modified by exploiting the ability of the β-cyclodextrin cavity to form an inclusion complex with the hydrophobic molecule adamantane. PEGylated adamantane derivatives were synthesized with and without the anisamide-targeting ligand on the terminal end of the PEG chain. Anisamide is known to bind specifically to the sigma receptor which is overexpressed on the surface of prostate cancer cells. The resulting nanocomplexes were slightly cationic and less than 300 nm in size. They successfully protected siRNA from serum-induced nuclease degradation and were non-toxic to prostate cancer cells. In addition, the targeted nanoparticles mediated high levels of siRNA cellular uptake and corresponding PLK1 gene knockdown in prostate cancer cells in vitro. To our knowledge, this is the first time that the ability of cyclodextrins to form inclusion complexes with adamantane derivatives has been exploited for the targeted delivery of siRNA to prostate cancer cells via the sigma receptor.
Keywords: Gene delivery; Prostate cancer; Anisamide targeted cyclodextrin; Targeted nanoparticle; Sigma receptor; Inclusion complex formation;

Self-assembled Micelle Loading Cabazitaxel for therapy of Lung Cancer by Bo Zhuang; Liang Du; Hongxia Xu; Xuelian Xu; Cheng Wang; Yingfang Fan; Mengyi Cong; Jiaqi Yin; Hongxia Li; Huashi Guan (146-155).
Display OmittedCBZ-PM can induce the Lewis lung carcinoma (LLC) cells death through G2/M arrest, which should be a potential anti-lung cancer agent.Lung cancer is a leading cause of cancer deaths worldwide, chemotherapy has improved overall survival but remains limited at <12 months median overall survival. Cabazitaxel is hopeful to do the same in advanced lung cancer as well as in metastatic prostate cancer. However, its clinical application was restricted due to its high hydrophobicity and severe side effects. To overcome these problems, we developed self-assembled micelle loading cabazitaxel (CBZ-PM) for therapy of lung cancer. The CBZ-PM has high drug loading (10.52%) and encapsulation efficiency (99.30%) with particle size of 28.77 ± 0.52 nm and polydisperse index of 0.114 ± 0.012. The transmission electron microscope image presented its spherical and homogeneous appearance. In vitro release profile showed CBZ-PM has a sustained-release behavior. Furthermore, the result of cell proliferation assays proved that CBZ-PM could induce the Lewis lung carcinoma (LLC) cells death through G2/M arrest more effectively than free CBZ. In vivo anti-tumor activity of CBZ-PM was further studied in mice model of LLC. The tumor inhibitory rate of CBZ-PM was more than 50% and the survival time of LLC bearing mice was efficiently prolonged following administration of CBZ-PM. In addition, the immunohistochemical study showed that more apoptosis cells were detected in the tumor tissue of CBZ-PM group than that of the positive control group. All these indicated that CBZ-PM served as a potential anti-lung cancer agent.
Keywords: Cabazitaxel; Polymeric micelle; Antitumor; Lewis lung carcinoma;

Display OmittedIn spite of intense efforts in the last 20 years, the current state of affairs regarding evaluation of adequacy of pharmaceutical mixing is at an impressive standstill, characterized by two draft guidances, one withdrawn, and the other never approved. We here analyze the regulatory, scientific and technological situation and suggest a radical, but logical approach calling for a paradigm shift regarding sampling of pharmaceutical blends. In synergy with QbD/PAT efforts, blend uniformity testing should only be performed with properly designed sampling that can guarantee representativity—in contrast to the current deficient thief sampling. This is necessary for suitable in-process specifications and dosage units meeting desired specifications. The present exposé shows how process sampling based on the Theory of Sampling (TOS) constitutes a new asset for regulatory compliance, providing procedures that suppress hitherto adverse sampling errors. We identify that the optimal sampling location is after emptying the blender, guaranteeing complete characterisation of the residual heterogeneity. TOS includes variographic analysis that decomposes the effective total sampling and analysis error (TSE + TAE) from the variability of the manufacturing process itself. This approach provides reliable in-process characterization allowing independent approval or rejection by the Quality Control unit. The science-based sampling principles presented here will facilitate full control of blending processes, including whether post-blending segregation influences the material stream that reaches the tabletting feed-frame.
Keywords: Theory of Sampling; TOS; Blend uniformity analysis (BUA); Variographic analysis; Measurement uncertainty; Sampling paradigm shift; Regulatory science; Quality control;

A novel hot-melt extrusion formulation of albendazole for increasing dissolution properties by Laura Martinez-Marcos; Dimitrios A. Lamprou; Roy T. McBurney; Gavin W. Halbert (175-185).
Display OmittedThe main aim of the research focused on the production of hot-melt extrusion (HME) formulations with increased dissolution properties of albendazole (ABZ). Therefore, HME was applied as a continuous manufacturing technique to produce amorphous solid dispersions of the poorly water soluble drug ABZ combined with the polymer matrix polyvinylpyrrolidone PVP K12. HME formulations of ABZ–PVP K12 comprised a drug content of 1%, 5% and 10% w/w. The main analytical characterisation techniques used were scanning electron microscopy (SEM), micro-computed tomography (μ-CT), X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC) and dissolution profile studies. The application of SEM, XRPD and DSC evidenced drug physical transformation from crystalline to amorphous state and therefore, the achievement of an amorphous solid dispersion. The introduction of a novel technique, μ-CT, to characterise the internal structure of these materials revealed key information regarding materials distribution and void content. Dissolution profile studies evidenced a high increase in drug release profile compared to pure ABZ. These promising results can lead to a great enhancement of the oral bioavailability of ABZ dosage forms. Therefore, HME is a potential continuous manufacturing technique to overcome ABZ poor solubility properties and lead to a significant increase in the therapeutic effect.
Keywords: Hot-melt extrusion; Amorphous solid dispersions; Albendazole; Continuous manufacturing; μ-CT;

Host-mediated Leishmania donovani treatment using AR-12 encapsulated in acetalated dextran microparticles by M.A. Collier; K.J. Peine; S. Gautam; S. Oghumu; S. Varikuti; H. Borteh; T.L. Papenfuss; A.R. Sataoskar; E.M. Bachelder; K.M. Ainslie (186-194).
Display OmittedLeishmaniasis is a disease caused by parasites of Leishmania sp., which effects nearly 12 million people worldwide and is associated with treatment complications due to widespread parasite resistance toward pathogen-directed therapeutics. The current treatments for visceral leishmaniasis (VL), the systemic form of the disease, involve pathogen-mediated drugs and have long treatment regimens, increasing the risk of forming resistant strains. One way to limit emergence of resistant pathogens is through the use of host-mediated therapeutics. The host-mediated therapeutic AR-12, which is FDA IND-approved for cancer treatment, has shown activity against a broad spectrum of intracellular pathogens; however, due to hydrophobicity and toxicity, it is difficult to reach therapeutic doses. We have formulated AR-12 into microparticles (AR-12/MPs) using the novel biodegradable polymer acetalated dextran (Ace-DEX) and used this formulation for the systemic treatment of VL. Treatment with AR-12/MPs significantly reduced liver, spleen, and bone marrow parasite loads in infected mice, while combinatorial therapies with amphotericin B had an even more significant effect. Overall, AR-12/MPs offer a unique, host-mediated therapy that could significantly reduce the emergence of drug resistance in the treatment of VL.
Keywords: AR-12; Visceral leishmaniasis; Dose sparing; Drug delivery; Microparticles;

Display OmittedThe elevated expression of bombesin receptors in many of the deadliest cancers has attracted special interest in developing bombesin-directed agents for tumor imaging and therapy. Previously, we constructed the chimeric peptide BB28 by fusing bombesin to a mitochondria-disrupting peptide. BB28 selectively induced the apoptosis of various tumor cells in vitro and showed promising in vivo antitumor effects. In general, a short circulating half-life limits the in vivo effect of peptides. To prolong the half-life of BB28, here, we generated the novel peptide ABB28 by fusing an albumin-binding domain (ABD) to the N-terminus of BB28. ABB28 exhibited much higher binding affinity for albumin than BB28, and this modification extended the peptide half-life from several minutes to 2 h. Optical imaging revealed that ABB28 accumulated in xenografted tumors within 1 h post-injection and persisted at an evident level for up to 24 h. ABB28 exerted stronger tumor-suppressive effects than BB28. Significant differences in the tumor volumes (P  < 0.001) and the tumor weights (P  = 0.002) were observed between ABB28- and BB28-treated mice. Moreover, ABB28 exhibited tumor suppression comparable to that of PEGylated 5K-BB28 in vivo. These results suggest that half-life extension via ABD fusion represents a useful strategy for optimizing bombesin-directed pharmaceuticals for cancer-targeted therapy.
Keywords: Tumor-homing peptide; Bombesin; Albumin; Albumin-binding domain; Drug carrier;

Micronization, characterization and in-vitro dissolution of shellac from PGSS supercritical CO2 technique by Philip W. Labuschagne; Brendon Naicker; Lonji Kalombo (205-216).
Display OmittedThe purpose of this investigation was to determine whether shellac, a naturally occurring material with enteric properties, could be processed in supercritical CO2 (sc-CO2) using the particles from gas saturated solution (PGSS) process and how process parameters affect the physico-chemical properties of shellac. In-situ attenuated total reflection fourier transform infra-red (ATR-FTIR) spectroscopy showed that CO2 dissolves in shellac with solubility reaching a maximum of 13% (w/w) at 300 bar pressure and 40 °C and maximum swelling of 28%. The solubility of sc-CO2 in shellac allowed for the formation of porous shellac structures of which the average pore diameter and pore density could be controlled by adjustment of operating pressure and temperature. In addition, it was possible to produce shellac microparticles ranging in average diameter from 180 to 300 μm. It was also shown that processing shellac in sc-CO2 resulted in accelerated esterification reactions, potentially limiting the extent of post-processing “ageing” and thus greater stability. Due to additional hydrolysis reactions enhanced by the presence of sc-CO2, the solubility of shellac at pH 7.5 was increased by between 4 and 7 times, while dissolution rates were also increased. It was also shown that the in-vitro dissolution profiles of shellac could be modified by slight adjustment in operating temperatures.
Keywords: Shellac; Supercritical carbon dioxide; Processing; In-situ foaming; Micronization; Dissolution;

Incorporation of acetaminophen as an active pharmaceutical ingredient into porous lactose by Amirali Ebrahimi; Morteza Saffari; Fariba Dehghani; Timothy Langrish (217-227).
Display OmittedA new formulation method for solid dosage forms with drug loadings from 0.65 ± 0.03% to 39 ± 1% (w/w) of acetaminophen (APAP) as a model drug has been presented. The proposed method involves the production of highly-porous lactose with a BET surface area of 20 ± 1 m2/g as an excipient using a templating method and the incorporation of drug into the porous structure by adsorption from a solution of the drug in ethanol. Drug deposition inside the carrier particles, rather than being physically distributed between them, eliminated the potential drug/carrier segregation, which resulted in excellent blend uniformities with relative standard deviations of less than 3.5% for all drug formulations. The results of DSC and XRD tests have shown deposition of nanocrystals of APAP inside the nanopores of lactose due the nanoconfinement phenomenon. FTIR spectroscopy has revealed no interaction between the adsorbed drug and the surface of lactose. The final loaded lactose particles had large BET surface areas and high porosities, which significantly increased the crushing strengths of the produced tablets. In vitro release studies in phosphate buffer (pH 5.8) have shown an acceptable delivery performance of 85% APAP release within 7 minutes for loaded powders filled in gelatin capsules.
Keywords: Content uniformity; Drug loading; Nanoconfinement; Acetaminophen; Spray drying; Porous lactose;

The influence of microemulsion structure on their skin irritation and phototoxicity potential by Branka Rozman; Mirjam Gosenca; Françoise Falson; Mirjana Gašperlin (228-235).
Display OmittedThe purpose of this study was to examine skin irritation and phototoxicity potentials of several microemulsions (ME), all comprising approximately the same percentage of surfactant mixture, but varying oil/water content and consequently inner structure being either droplet-like (o/w ME, o/w ME carbomer, w/o ME and w/o ME white wax) or lamellar (gel-like ME). Two different in vitro methods were used: MTT assay (performed either on reconstructed human epidermis (RHE) or NCTC 2544 cells) and pig ear test. Neither assay revealed the difference among ME with droplet-like structure. Then again, pig ear test and MTT assay performed on RHE indicated that gel-like ME is more irritant compared to other tested ME, whereas no difference among formulations were observed by MTT assay on NCTC 2544 cells. The reasonable explanation is destruction and consequently uniform structure of ME upon dilution that is inevitable for testing on cell cultures. The results of phototoxicity test again indicated the increased potential of gel-like ME to cause adverse effects on skin. It can be concluded that for ME consisting of the same amount of identical surfactants but having different structure the latter represent a crucial factor that determines their dermal toxicity.
Keywords: Microemulsion; Structure; Skin irritation; RHE; Keratinocyte cell cultures; Phototoxicity;

Preparation and in vivo evaluation of insulin-loaded biodegradable nanoparticles prepared from diblock copolymers of PLGA and PEG by Yusuf Haggag; Yasser Abdel-Wahab; Opeolu Ojo; Mohamed Osman; Sanaa El-Gizawy; Mohamed El-Tanani; Ahmed Faheem; Paul McCarron (236-246).
Display OmittedThe aim of this study was to design a controlled release vehicle for insulin to preserve its stability and biological activity during fabrication and release. A modified, double emulsion, solvent evaporation, technique using homogenisation force optimised entrapment efficiency of insulin into biodegradable nanoparticles (NP) prepared from poly (dl-lactic-co-glycolic acid) (PLGA) and its PEGylated diblock copolymers. Formulation parameters (type of polymer and its concentration, stabiliser concentration and volume of internal aqueous phase) and physicochemical characteristics (size, zeta potential, encapsulation efficiency, in vitro release profiles and in vitro stability) were investigated. In vivo insulin sensitivity was tested by diet-induced type II diabetic mice. Bioactivity of insulin was studied using Swiss TO mice with streptozotocin-induced type I diabetic profile. Insulin-loaded NP were spherical and negatively charged with an average diameter of 200–400 nm. Insulin encapsulation efficiency increased significantly with increasing ratio of co-polymeric PEG. The internal aqueous phase volume had a significant impact on encapsulation efficiency, initial burst release and NP size. Optimised insulin NP formulated from 10% PEG–PLGA retained insulin integrity in vitro, insulin sensitivity in vivo and induced a sustained hypoglycaemic effect from 3 h to 6 days in type I diabetic mice.
Keywords: Insulin; PEG-PLGA; Nanoparticles; Stability; Controlled delivery; Diabetes;

Effect of dendrimers on selected enzymes—Evaluation of nano carriers by Maksim Ionov; Aliaksei Ihnatsyeu-Kachan; Sylwia Michlewska; Natallia Shcharbina; Dzmitry Shcharbin; Jean-Pierre Majoral; Maria Bryszewska (247-254).
Display OmittedIn the field of nanotechnology, dendrimers represent a new class of highly branched macromolecules that is receiving a stimulating and rising interest. The structural organization of these synthetic macromolecules provides promising opportunities for using them as nano-carriers of drugs or gene material to be delivered to the target cells. For applications of dendrimers as drug carriers, analysis of their specific interactions with biological structures at molecular level is very important. This paper describes the molecular interactions between cationic phosphorus dendrimers of third and fourth generation (CPD G3 and CPD G4) and 3 plasma regulatory proteins, namely aspartate transaminase, alkaline phosphatase and l-lactic dehydrogenase. Dendrimer–protein interactions were studied using spectrofluorimetric, circular dichroism and dynamic light scattering techniques. Their morphology in the presence or absence of dendrimers was examined by transmission electron microscopy. The results suggest that both dendrimers form positively charged complexes with HIV-derived peptides. The circular dichroism spectra show that these dendrimers can significantly change the secondary structure of proteins, indicating formation of protein/dendrimer complexes.
Keywords: Phosphorus dendrimer; Plasma regulatory proteins; Complex formation; Surface charge;

Strategic application of self-micellizing solid dispersion technology to respirable powder formulation of tranilast for improved therapeutic potential by Hiroki Suzuki; Yoshiki Kojo; Keisuke Yakushiji; Kayo Yuminoki; Naofumi Hashimoto; Satomi Onoue (255-262).
Display OmittedThe present study aimed to develop an inhalable self-micellizing solid dispersion of tranilast (SMSD/TL) using poly[MPC-co-BMA] to improve the therapeutic potential and safety.The safety of poly[MPC-co-BMA] in lungs was assessed using rat lung epithelium-derived L2 cells. SMSD/TL and respirable powder of SMSD/TL (SMSD/TL-RP) were prepared using a wet milling system and jet mill, respectively. The physicochemical properties of TL formulations were characterized in terms of dissolution, morphology, and particle size. Pharmacological and pharmacokinetic studies were also conducted on inhaled SMSD/TL-RP.The lactate dehydrogenase level from L2 cells treated with poly[MPC-co-BMA] was lower than that with polysorbate 80, a positive control. SMSD/TL showed enhanced dissolution behavior of TL. The jet milled SMSD/TL particles easily separated from the lactose carrier, and the particle size was suitable for inhalation. Compared with RP of TL, inhaled SMSD/TL-RP (100 μg-TL/rat) could more strongly suppress the inflammatory responses in antigen-sensitized rats. The TL level in plasma after intratracheal administration of SMSD/TL-RP at a pharmacological effective dose (100 μg-TL/rat) was ca. 4.2-fold lower than that after oral administration of TL solution at a clinical dose (1.67 mg/kg).SMSD/TL-RP might be an attractive dosage form to improve the anti-inflammatory effects and safety of TL.
Keywords: Tranilast; Poly[MPC-co-BMA]; Respirable powder; Anti-inflammatory effect; Self-micellizing solid dispersion;

Multimodal approach to characterization of hydrophilic matrices manufactured by wet and dry granulation or direct compression methods by Piotr Kulinowski; Krzysztof Woyna-Orlewicz; Jadwiga Obrał; Gerd-Martin Rappen; Dorota Haznar-Garbacz; Władysław P. Węglarz; Renata Jachowicz; Gabriela Wyszogrodzka; Jolanta Klaja; Przemysław P. Dorożyński (263-270).
Display OmittedThe purpose of the research was to investigate the effect of the manufacturing process of the controlled release hydrophilic matrix tablets on their hydration behavior, internal structure and drug release. Direct compression (DC) quetiapine hemifumarate matrices and matrices made of powders obtained by dry granulation (DG) and high shear wet granulation (HS) were prepared. They had the same quantitative composition and they were evaluated using X-ray microtomography, magnetic resonance imaging and biorelevant stress test dissolution.Principal results concerned matrices after 2 h of hydration: (i) layered structure of the DC and DG hydrated tablets with magnetic resonance image intensity decreasing towards the center of the matrix was observed, while in HS matrices layer of lower intensity appeared in the middle of hydrated part; (ii) the DC and DG tablets retained their core and consequently exhibited higher resistance to the physiological stresses during simulation of small intestinal passage than HS formulation.Comparing to DC, HS granulation changed properties of the matrix in terms of hydration pattern and resistance to stress in biorelevant dissolution apparatus. Dry granulation did not change these properties—similar hydration pattern and dissolution in biorelevant conditions were observed for DC and DG matrices.
Keywords: Magnetic Resonance Imaging (MRI); X-ray microtomography (μCT, Micro-CT); Biorelevant stress test dissolution; Quetiapine controlled release tablets; Dry granulation; High shear granulation; Direct compression;

Micropellet-loaded rods with dose-independent sustained release properties for individual dosing via the Solid Dosage Pen by Eva Julia Laukamp; Klaus Knop; Markus Thommes; Joerg Breitkreutz (271-279).
Display OmittedIndividual dosing of medicines is relevant for paediatrics, geriatrics and personalised medicine. The Solid Dosage Pen (SDP) allows for individual dosing by cutting monolithic, tablet-like drug carriers of pre-defined heights.The aim of the present study was to develop micropellet-loaded rods (MPLRs) with dose-independent sustained release properties for individual dosing via the Solid Dosage Pen. Therefore, micropellets were successfully layered with carbamazepine and coated with polyvinyl acetate (PVAc) and PVAc/polyvinyl alcohol-polyethylene glycol (PVA-PEG). The tensile strength of the sustained release micropellets (300–450 μm) was more than two times higher (12.6–17.1 MPa) than pressures occurring during ram-extrusion of the MPLRs (below 5.8 MPa). Due to relative crystallinities above 93% for PVAc and PVA-PEG a low solubility of the coating films within the PEG-matrix was observed. The sustained release micropellets were successfully incorporated into MPLRs. Drug release properties of the pellets maintained after embedding into the matrix. Hence, the MPLRs provided dose-independent prolonged drug liberation which was not achieved for drug-loaded rods before. The MPLRs permitted the application of the SDP with a precise drug delivery from individually cut single doses. Storage stability was proven for MPLRs containing PVAc/PVA-PEG coated pellets. In conclusion, the MPLRs combined the advantages of multiparticulate dosage forms with the SDP as a device for individual dosing.
Keywords: Individual dosing; Solid Dosage Pen; Multiparticulate dosage form; Micropellets; Sustained release;

Display OmittedThe objective of this study was to develop solid solution (SSL) using hot-melt extrusion as a continuous manufacturing method. Powder blends of artesunate (ARS) a water insoluble drug with either Soluplus (SOL) or Kollidon VA64 (VA64) and additives in the form of surfactants or plasticizers were extruded to manufacture extrudes. The incorporation of surfactant or plasticizers facilitates smooth extrusion processing of the drug-excipient blend which directly reduced the residence time to form extrudes and works as parameter to control flow of the drug-excipients melt inside the extruder barrel. Differential scanning calorimetry (DSC) and X-ray diffraction (TXRD) analysis revealed the existence of the drug within the extrudes in amorphous state. Scanning electron microscopy (SEM), Raman spectroscopy (RS), Raman imaging (RI) and Atomic force microscopy (AFM) analytical characterization were carry out on the SSL formulations showed a homogeneous drug distribution within the extrudes. 2D NMR and 1H NMR studies were undertaken to reveal the possible drug–excipient interactions. The SSL produced via continuous HME processing showed significantly faster release of ARS compared to the pure drug substance. It is observed that F1 SSL (soluplus based) have 66.44 times higher AUC(0–72) and 16.60 times higher C max than pure ARS. Also K1 SSL (Kollidon VA64 based) have 62.20 times higher AUC(0–72) and 13.40 times higher C max than pure ARS.
Keywords: Artesunate; Atomic force microscopy; Bioavailability; Hot melt extrusion; solid solution; 2D NMR; Dissolution;

Evaluation of the efficiency and safety in cosmetic products by Meryem Uckaya; Fatih Uckaya; Nazan Demir; Yasar Demir (295-300).
Chemicals used in cosmetics have to interact with enzymes for beneficial or destroy purpose after they enter in our body. Active sections of enzymes that catalyze reactions have three dimensions and they are active optically. When these limitations of catalytic sections are considered, it may be considered that defining geometric specifications of chemical materials and functional groups they contain may contribute on safety evaluations of cosmetic products.In this study, defining similarities and differences of geometric structures of chemicals that are prohibited to be used in cosmetic products and chemical that are allowed to be used by using group theory and analyze of functional groups that are often encountered in these chemicals are aimed.Molecule formulas related to chemical material of, 276 pieces chemicals that are prohibited to be used in cosmetic products and 65 pieces chemicals that are allowed, are used as the material. Two and three-dimension structures of these formulas are drawn and types and quantity of functional groups they contain are defined.And as a method, freeware (Free Trial) version of “Chem-BioOffice Ultra 13.0 Suite” chemical drawing program to draw two and three-dimension of formulas, “Campus-Licensed” version that are provided for use by our university of “Autodesk 3DS Max” for three-dimension drawings are used. In order to analyze geometric specifications of drawn molecules according to Group Theory and define type and quantity of available functional groups, Excel applications developed by Prof. Dr. Yaşar Demir are used.
Keywords: Cosmetic; Three-dimensional structure; Enzyme; Group theory; Safety;

Nanostructured lipid carrier mediates effective delivery of methotrexate to induce apoptosis of rheumatoid arthritis via NF-κB and FOXO1 by Neeraj K. Garg; Rajeev K. Tyagi; Bhupinder Singh; Gajanand Sharma; Pradip Nirbhavane; Varun Kushwah; Sanyog Jain; Om Prakash Katare (301-320).
Display OmittedPresent study was designed to develop novel nano-structured lipid carriers (NLCs) formulated by lipid mixture and chemical permeation enhancer-based hydrogel for an effective transdermal delivery of methotrexate (MTX). The prepared NLCs were optimized with different preparative variables such as particle size <200 nm, poly-dispersity index (PDI) <0.2, and entrapment efficiency ∼85%. The drug incorporated into NLCs-gel base showed excellent spread ability without any grittiness during rheological behavior and texture profile analysis. The in vitro release showed biphasic release pattern with initial fast release of drug (>50%) in 8 h followed by sustained release (up to 85%) by the end of 48th h. NLCs showed greater uptake in human hyper-proliferative keratinocyte cell line (HaCaT). NLCs showed increased expression of inflammatory mediators as well asapoptosis in U937 monocytic cells. The greater expression of pro-apoptotic gene Bim regulated by NF-κB-IkB and FOXO1 is supported by fold regulations calculated for various apoptotic and pro-inflammatory biomarkers carried out by RT-PCR. The immunocytochemistry to detect IL-6 expression and immunofluorescence assay suggested that induced apoptosis occurs in experimentally induced in vitro arthritis model treated with NLCs-MTX. We saw reduced inflammation and triggered apoptosis through NF-κB & FOXO1 pathways induced by MTX loaded NLCs in rheumatoid arthritic cells. In addition, formulated NLCs exhibit better skin permeation with higher permeation flux & enhancement ratio as shown by confocal laser scanning microscopy (CLSM). Moreover, histopathological examinations of skin are suggestive of safety potential of NLCs.
Keywords: Nanostructured lipid carriers (NLCs); Methotrexate; Rheumatoid Arthritis; Apoptosis; NF-κB-IkB; FOXO1; RT-PCR; α-Terpineol; Phospholipids S-100; Gelucire 50/13; Transcutol P; Transdermal delivery;

Characterization of a food-based enteric coating for capsules and its compatibility with an alternative sealing method by Romain Caillard; Jean-Simon Blais; Gérard Akinocho; Wilfrid Jacques (321-329).
Display OmittedEfficiency of a new protein-based enteric coating for capsules was studied. Coating physical–chemical properties were compared to those obtained from a well-known methacrylate-based enteric coating (Eudragit®). Swelling in simulated gastric fluid (SGF) was 20 times higher than for Eudragit® films. Mechanical properties (elastic modulus, elongation and puncture strength at break) were comparable to those measured from a standard Eudragit® formulation.Pilot-scale coating trials were performed following three methods: using a standard spray-gun configuration, using a HPC-based seal-coat prior to enteric coating and using an “inverted” spray-gun configuration. The effect of these methods on capsules sealing and in vitro gastric performance was studied. In vitro tests were performed following the two USP official methods: disintegration and dissolution.Inverted gun configuration and HPC-sealing showed the highest sealing efficiency and the best in vitro performance. Capsules with a weight gain of 14–16% generally passed all USP tests (no disintegration evidence after 60 min in SGF; release below 10% after 2 h of experiments in SGF). However, in some cases, slight differences between results obtained from dissolution and disintegration tests were pointed out.This work demonstrates the potential of a protein-based enteric coating and underlines the importance of capsules sealing.
Keywords: Food proteins; Enteric coating; Sealing; Capsules; Gastric protection; Scale-up;

Display OmittedThe impact of hydrodynamics and media composition on nifedipine dissolution profile from IR (immediate release) soft capsules was investigated using dissolution apparatus USP1, USP2, USP3 and USP4 (United State Pharmacopoeia). Media composition was varied in terms of pH and content, to mimic the dosage form intake with water or non-alcoholic beverages (orange juice) and alcoholic beverages (orange juice/ethanol mixture (47% v/v)). Through construction of in vitro–in vivo correlations (IVIVC) with corresponding in vivo data from the literature, it was possible to evaluate the in vitro conditions that are likely to simulate the in vivo formulation behaviour. Both linear and nonlinear correlations were obtained depending on experimental set-ups. Testing of 20 mg nifedipine capsules in FaSSGFst (Fasted State Simulated Gastric Fluid pH 1.6; water administration) produced IVIVC with the USP3 (after time scaling) and USP4 apparatus. IVIVC were obtained for USP2, USP3 and USP4 in FaSSGFoj (Fasted State Simulated Gastric Fluid pH 3.4; orange juice administration). Linear and nonlinear correlations were obtained with the USP1, USP2 and USP3 apparatus when testing the capsules in FaSSGFoj/EtOH (orange juice/ethanol administration). This study highlighted that selection of physiologically relevant dissolution set-ups is critical for predicting the in vivo impact of formulations co-administration with water, non-alcoholic and alcoholic beverages.
Keywords: IVIVC; Biorelevant dissolution; Capsule rupture time; Nifedipine; Immediate release; Hydrodynamics; Special dissolution media; Non-alcoholic beverages; Alcoholic beverages;

A UV-Raman spectrometry method for quality control of anticancer preparations: Results after 18 months of implementation in hospital pharmacy by Flore Nardella; Morgane Beck; Pierre Collart-Dutilleul; Guillaume Becker; Coralie Boulanger; Laurent Perello; Anne Gairard-Dory; Bénédicte Gourieux; Geneviève Ubeaud-Séquier (343-350).
Display OmittedIn France, chemotherapy preparation units of hospital pharmacy compound cytotoxic infusion bags adapted to each patient. The narrow therapeutic index of these preparations led us to implement qualitative and quantitative control for patients’ safety. To this aim, we calibrated an equipment combining UV–vis spectrometry and Raman spectroscopy (QC Prep+®) and monitored 14 different molecule–solvent combinations over a 18 months period. This rapid and specific method allowed the qualitative and quantitative analysis of 1 mL sample tests in less than 2 min. On 5742 anticancer preparations, we obtained accepted results with more than 99.4% solvent identification, 99.6% drug identification and only 1.52% of preparations not matching quantitative specifications (±15% of theoretical concentration). This quantitative control enabled us to pinpoint some critical points of production for two of the most common preparations. We thus updated the procedures of reconstitution and preparation, increasing the quality of final product. UV-Raman spectrometry is thus an effective tool to control chemotherapy infusions and to improve good practices of preparation.
Keywords: UV-Raman; Anticancer preparation; Chemical control; Hospital pharmacy;

Hydrophobic ion pairing of a minocycline/Ca2+/AOT complex for preparation of drug-loaded PLGA nanoparticles with improved sustained release by Alexander Dontsios Holmkvist; Annika Friberg; Ulf J. Nilsson; Jens Schouenborg (351-357).
Display OmittedPolymeric nanoparticles is an established and efficient means to achieve controlled release of drugs. Incorporation of minocycline, an antibiotic with anti-inflammatory and neuroprotective properties, into biodegradable nanoparticles may therefore provide an efficient means to combat foreign body reactions to implanted electrodes in the brain. However, minocycline is commonly associated with poor encapsulation efficiencies and/or fast release rates due to its high solubility in water. Moreover, minocycline is unstable under conditions of low and high pH, heat and exposure to light, which exacerbate the challenges of encapsulation. In this work drug loaded PLGA nanoparticles were prepared by a modified emulsification-solvent-diffusion technique and characterized for size, drug encapsulation and in vitro drug release. A novel hydrophobic ion pair complex of minocycline, Ca2+ ions and the anionic surfactant AOT was developed to protect minocycline from degradation and prolong its release. The optimized formulation resulted in particle sizes around 220 nm with an entrapment efficiency of 43% and showed drug release over 30 days in artificial cerebrospinal fluid. The present results constitute a substantial increase in release time compared to what has hitherto been achieved for minocycline and indicate that such particles might provide useful for sustained drug delivery in the CNS.
Keywords: Minocycline; Poly(d,l-lactic-co-glycolic acid) (PLGA); Nanoparticles; Drug release; Hydrophobic ion paring; Emulsification-solvent-diffusion method;

Strategies for encapsulation of small hydrophilic and amphiphilic drugs in PLGA microspheres: State-of-the-art and challenges by Farshad Ramazani; Weiluan Chen; Cornelis F. van Nostrum; Gert Storm; Fabian Kiessling; Twan Lammers; Wim E. Hennink; Robbert J. Kok (358-367).
Display OmittedPoly(lactide-co-glycolide) (PLGA) microspheres are efficient delivery systems for controlled release of low molecular weight drugs as well as therapeutic macromolecules. The most common microencapsulation methods are based on emulsification procedures, in which emulsified droplets of polymer and drug solidify into microspheres when the solvent is extracted from the polymeric phase. Although high encapsulation efficiencies have been reported for hydrophobic small molecules, encapsulation of hydrophilic and/or amphiphilic small molecules is challenging due to the partitioning of drug from the polymeric phase into the external phase before solidification of the particles. This review addresses formulation-related aspects for efficient encapsulation of small hydrophilic/amphiphilic molecules into PLGA microspheres using conventional emulsification methods (e.g., oil/water, water/oil/water, solid/oil/water, water/oil/oil) and highlights novel emulsification technologies such as microfluidics, membrane emulsification and other techniques including spray drying and inkjet printing. Collectively, these novel microencapsulation technologies afford production of this type of drug loaded microspheres in a robust and well controlled manner.
Keywords: Polymeric microspheres; PLGA; Microencapsulation; Encapsulation efficiency; Sustained release;

Synthesis and characterisation of mucoadhesive thiolated polyallylamine by Sarah Duggan; Helen Hughes; Eleanor Owens; Elaine Duggan; Wayne Cummins; Orla O’ Donovan (368-375).
Display OmittedThe thiolation of polyallylamine (PAAm) for use in mucoadhesive drug delivery has been achieved. PAAm was reacted with different ratios of Traut’s reagent, yielding products with thiol contents ranging from 134–487 μmol/g. Full mucoadhesive characterisation of the thiolated PAAm samples was conducted using swelling studies, mucoadhesive testing on porcine intestinal tissue and rheology. Both swelling and cohesive properties of the thiolated PAAm products were vastly improved in comparison to an unmodified PAAm control. The swelling abilities of the thiolated samples were high and the degree of thiolation of the products affected the initial rate of swelling. High levels of mucoadhesion were demonstrated by the thiolated PAAm samples, with adhesion times of greater than 24 h measured for all three samples and, thus, thiol content did not appear to influence mucoadhesion. Rheological studies of the thiolated PAAm samples showed an increase in G′ and G″ values upon the addition of a mucin solution which was not observed in the unmodified control, again highlighting the mucoadhesive interactions between these thiolated polymers and mucin. The synthesis of thiolated PAAm by reaction with Traut’s reagent and resulting mucoadhesive properties demonstrates its potential for use a mucoadhesive drug delivery device.
Keywords: Thiolation; Polyallylamine; Traut’s reagent; Mucoadhesion;

Display OmittedThree-dimensional printing includes a wide variety of manufacturing techniques, which are all based on digitally-controlled depositing of materials (layer-by-layer) to create freeform geometries. Therefore, three-dimensional printing processes are commonly associated with freeform fabrication techniques. For years, these methods were extensively used in the field of biomanufacturing (especially for bone and tissue engineering) to produce sophisticated and tailor-made scaffolds from patient scans. This paper aims to review the processes that can be used in pharmaceutics, including the parameters to be controlled. In practice, it not straightforward for a formulator to be aware of the various technical advances made in this field, which is gaining more and more interest. Thus, a particular aim of this review is to give an overview on the pragmatic tools, which can be used for designing customized drug delivery systems using 3D printing.
Keywords: 3D printing; Pharmaceutics; Drug delivery systems; Solid freeform fabrication; Rapid prototyping;

Continuous preparation of polymer coated drug crystals by solid hollow fiber membrane-based cooling crystallization by Dengyue Chen; Dhananjay Singh; Kamalesh K. Sirkar; Robert Pfeffer (395-402).
Display OmittedA facile way to continuously coat drug crystals with a polymer is needed in controlled drug release. Conventional polymer coating methods have disadvantages: high energy consumption, low productivity, batch processing. A novel method for continuous polymer coating of drug crystals based on solid hollow fiber cooling crystallization (SHFCC) is introduced here. The drug acting as the host particle and the polymer for coating are Griseofulvin (GF) and Eudragit RL100, respectively. The polymer’s cloud point temperature in its acetone solution was determined by UV spectrophotometry. An acetone solution of the polymer containing the drug in solution as well as undissolved drug crystals in suspension were pumped through the tube side of the SHFCC device; a cold liquid was circulated in the shell side to rapidly cool down the feed solution-suspension in the hollow-fiber lumen. The polymer precipitated from the solution and coated the suspended crystals due to rapid temperature reduction and heterogeneous nucleation; crystals formed from the solution were also coated by the polymer. Characterizations by scanning electron microscopy, thermogravimetric analysis, laser diffraction spectroscopy, X-ray diffraction, Raman spectroscopy, and dissolution tests show that a uniformly coated, free-flowing drug/product can be obtained under appropriate operating conditions without losing the drug’s pharmaceutical properties and controlled release characteristics.
Keywords: Polymer coating; Drug crystals; Membrane-based cooling crystallization; Solid hollow fiber;

New insights into eutectic cream skin penetration enhancement by Sarah Fiala; Marie Roman; Ricardo Inacio; Sumaia Mashal; Marc B. Brown; Stuart A. Jones (403-411).
Molecular arrangement in ELMA.Display OmittedThe manner in which the eutectic cream EMLA® enhances the percutaneous penetration of lidocaine and prilocaine into human skin is still not fully understood. The purpose of this study was to investigate if the modification of drug aggregation played a role in the way EMLA® facilitates delivery. Light scattering analysis of lidocaine alone in water gave a critical aggregation concentration (CAC) of 572 μM and a mean aggregate size of 58.8 nm. The analysis of prilocaine in identical conditions gave a CAC of 1177 μM and a mean aggregate size of 105.7 ± 24.8 nm. When the two drugs were mixed at their eutectic 1:1 ratio in water the CAC reduced to 165.8 μM and the aggregate size was 43.82 nm. This lidocaine–prilocaine interaction in water was further modified upon addition of polyoxyethylene hydrogenated castor oil, the surfactant in the EMLA aqueous phase, to produce aggregates of <20 nm. The physical characterisation data suggested that it was the EMLA cream’s surfactant that modified the drug molecular interactions in the aqueous continuous phase and caused a 6 fold higher drug penetration through human epidermal tissue compared to the oil formulations tested in this study.
Keywords: EMLA®; Lidocaine; Prilocaine; Eutectic; Penetration enhancement; Skin;