International Journal of Pharmaceutics (v.492, #1-2)

Hot-melt extrusion of polyvinyl alcohol for oral immediate release applications by W. De Jaeghere; T. De Beer; J. Van Bocxlaer; J.P. Remon; C. Vervaet (1-9).
Display OmittedThe primary purpose of this study was to process partially hydrolyzed PVOH grades (degree of hydroxylation (DH): 33–88%) via HME and to evaluate them as carrier for oral immediate release dosage forms in order to improve the release rate of poorly water soluble drugs (i.e., HCT and CEL) via the formulation of solid dispersions. PVOH grades (DH >70%) were able to solubilize HCT and CEL up to 15%, but required higher extrusion temperature, due to the crystalline nature of PVOH. The highest drug release rate was observed from hot-melt extruded PVOH samples with a high DH. While drug release from extrudates consisting of PVOH with a low DH was affected by ionic strength, there was no influence of pH and ionic strength on HCT release from PVOH samples with a higher DH. However, PVOH (DH >70%) required higher extrusion temperatures, which could hamper its application for thermosensitive drugs. Therefore, the secondary purpose was to investigate the effect of sorbitol, a water-soluble plasticizer, on the thermal properties of hot-melt extruded PVOH (DH >70%). The melting of PVOH/sorbitol mixture was required to establish molecular interactions between PVOH and sorbitol. These molecular interactions were reflected in the HME behavior: whereas an extrusion temperature of 180 °C was necessary to process physical mixtures of PVOH (DH >70%) and sorbitol, only 140 °C was necessary during re-extrusion (after quench cooling and cryomilling) of the PVOH/sorbitol mixture. In addition, the in vitro and in vivo dug release of plasticized PVOH was examined; whereas the CEL/PVO/sorbitol system was able to maintain supersaturation during in vitro dissolution (0.1 N HCl) compared to Celebrex®, the in vivo pharmacokinetic parameters (AUC0–24 h, C max and T max) were highly comparable.
Keywords: Oral drug delivery; Hot-melt extrusion; Polyvinyl alcohol; Immediate release; Supersaturation;

Improved antitumor effect of paclitaxel administered in vivo as pH and glutathione-sensitive nanohydrogels by Elena Pérez; Ana Martínez; César Teijón; Rosa Olmo; José María Teijón; María Dolores Blanco (10-19).
Scheme of the antitumor activity and nanotoxicology experiments done in the present work. Stimuli- responsive nanohydrogels were evaluated as PTX nanocarriers in female athymic nude mice bearing HeLa human tumor xenografts at a single dose of 30 mg/kg.Display OmittedMost antitumor drugs usually affect not only rapidly dividing cells, such as those in tumors, but also highly proliferative cells in normal tissues. This nonspecific drawback could be successfully solved by using nanocarriers as controlled drug delivery systems. In this work, pH and redox-responsive nanohydrogels (NG) based on N-isopropylacrilamide (NIPA), N-hydroxyethyl acrylamide (HEEA) 2-acrylamidoethyl carbamate (2AAECM) and N,N′-cystaminebisacrylamide (CBA) as crosslinker were evaluated as bioreducible paclitaxel (PTX) nanocarriers for improving the accumulation of the drug within the tumor tissue and avoiding its conventional side effects. A single dose of PTX solution, unloaded-NHA 80/15/5CBA NG and PTX-loaded NHA 80/15/5-CBA NG (30 mg/kg PTX equivalent) were subcutaneously injected in female athymic nude mice bearing HeLa human tumor xenografts. PTX-loaded nanohydrogels showed higher antitumor activity than free PTX, as tumor evolution and Ki67 detection demonstrated. Histological tumor images revealed a higher content of defective mitotic figures and apoptotic bodies in PTX- treated tumors than in control or unloaded NG treated tumor samples. Nanohydrogels injection did not change any biochemical blood parameters, which means no liver or kidney damage after NG injection. However, differences in antioxidant defenses in MPS systems (liver, kidney and spleen) were observed among treatments, which may indicate an oxidative stress response after PTX injection.
Keywords: Anti-tumor efficacy; In vivo toxicity; Paclitaxel; Stimuli responsive nanohydrogel; Histopathology; Oxidative stress;

Release mechanisms of tacrolimus-loaded PLGA and PLA microspheres and immunosuppressive effects of the microspheres in a rat heart transplantation model by Ryo Kojima; Takatsune Yoshida; Hiroaki Tasaki; Hiroyuki Umejima; Masashi Maeda; Yasuyuki Higashi; Shunsuke Watanabe; Naoto Oku (20-27).
Display OmittedThe objective of this study was to elucidate the release and absorption mechanisms of tacrolimus loaded into microspheres composed of poly(lactic-co-glycolic acid) (PLGA) and/or polylactic acid (PLA). Tacrolimus-loaded microspheres were prepared by the o/w emulsion solvent evaporation method. The entrapment efficiency correlated with the molecular weight of PLGA, and the glass transition temperature of PLGA microspheres was not decreased by the addition of tacrolimus. These results indicate that intermolecular interaction between tacrolimus and the polymer would affect the entrapment of tacrolimus in the microspheres. Tacrolimus was released with weight loss of the microspheres, and the dominant release mechanism of tacrolimus was considered to be erosion of the polymer rather than diffusion of the drug. The whole-blood concentration of tacrolimus in rats was maintained for at least 2 weeks after a single subcutaneous administration of the microspheres. The pharmacokinetic profile of tacrolimus following subcutaneous administration was similar to that following intramuscular administration, suggesting that the release and dissolution of tacrolimus, rather than the absorption of the dissolved tacrolimus, were rate-limiting steps. Graft-survival time in a heart transplantation rat model was prolonged by the administration of tacrolimus-loaded microspheres. The microsphere formulation of tacrolimus would be expected to precisely control the blood concentration while maintaining the immunosuppressive effect of the drug.
Keywords: Tacrolimus; PLGA; PLA; Microsphere; Controlled release; Heart transplantation;

Display OmittedPolymeric micelles that can deliver drug to intended sites of the eye have attracted much scientific attention recently. The aim of this study was to evaluate the aqueous-based formulation of drug-loaded polymeric micelles that hold significant promise for ophthalmic drug delivery. This study investigated the synergistic performance of mixed polymeric micelles made of linear and branched poly(ethylene oxide)-poly(propylene oxide) for the more effective encapsulation of lornoxicam (LX) as a hydrophobic model drug. The co-micellization process of 10% binary systems combining different weight ratios of the highly hydrophilic poloxamers; Synperonic® PE/P84, and Synperonic® PE/F127 and the hydrophobic poloxamine counterpart (Tetronic® T701) was investigated by means of photon correlation spectroscopy and cloud point. The drug-loaded micelles were tested for their solubilizing capacity towards LX. Results showed a sharp solubility increase from 0.0318 mg/mL up to more than 2.34 mg/mL, representing about 73-fold increase. Optimized formulation was selected to achieve maximum drug solubilizing power and clarity with lowest possible particle size, and was characterized by 1HNMR analysis which revealed complete encapsulation of the drug within the micelles. Further investigations by histopathological and confocal laser studies revealed the non-irritant nature and good corneal penetrating power of the proposed nano-formulation.
Keywords: Ocular drug delivery; Single and mixed nanomicellar systems; Lornoxicam; Confocal laser scanning microscopy; Histopathological studies;

Drug-in-cyclodextrin-in-liposomes: A novel drug delivery system for flurbiprofen by Lina Zhang; Qi Zhang; Xin Wang; Wenji Zhang; Congcong Lin; Fen Chen; Xinggang Yang; Weisan Pan (40-45).
Display OmittedA novel delivery system based on drug-cyclodextrin (CD) complexation and liposomes has been developed to improve therapeutic effect. Three different means, i.e., co-evaporation (COE), co-ground (GR) and co-lyophilization (COL) and three different CDs (β-CD, HP-β-CD and SBE-β-CD) were contrasted to investigate the characteristics of the end products. FP/FP-CD loaded liposomes were obtained by thin layer evaporation technique. Size, zeta potential and encapsulation efficiency were investigated by light scattering analysis and minicolumn centrifugation. Differential scanning calorimetry (DSC) and transmission electron microscopy (TEM) showed the amorphous form of complexes and spherical morphology of FP-HP-β-CD COE loaded liposomes. The pH 7.4 phosphate buffer solution (PBS) was selected as the medium for the in vitro release. Wistar rats were put into use to study the pharmacokinetic behavior in vivo. FP-HP-β-CD COE loaded liposomes showed the better physicochemical characters that followed the average particle size, polydispersity index, zeta potential and mean encapsulation efficiency 158 ± 10 nm, 0.19 ± 0.1, −12.4 ± 0.1 mW and 56.1 ± 0.5%, separately. The relative bioavailability of FP-HP-β-CD COE loaded liposomes was 420%, 201% and 402% compared with FP solution, FP-HP-β-CD and FP-liposomes, respectively. In conclusion, the novel delivery system improved the relative bioavailability of FP significantly and provided a perspective way for delivery of insoluble drugs.
Keywords: Flurbiprofen; Cyclodextrins; Liposomes; In vitro; In vivo; Bioavailability;

Size characterization of commercial micelles and microemulsions by Taylor dispersion analysis by Joseph Chamieh; Florian Davanier; Vincent Jannin; Frédéric Demarne; Hervé Cottet (46-54).
Display OmittedIn this work, Taylor dispersion analysis was applied to the measurement of micelles (or microdroplets) molecular diffusion coefficient in micellar (or microemulsion) systems based on neutral/anionic/cationic or zwitterionic surfactants. The choice of the micellar marker and the influence the surfactant/marker concentrations on this determination are studied. Experimental results are compared to those derived from the literature using other experimental techniques. Taylor dispersion analysis, experienced in narrow capillaries, was found to be an efficient and suitable method for micelle (or microdroplet) size measurement due to: the low sample consumption, the absence of filtration requirement of the sample, the broad range of size determination (with no lower limit down to angstroms), the simplicity of the protocol, the possibility to measure the viscosity of surfactant solutions in given conditions and the determination of the weight-average micelle hydrodynamic radius. Application to the size-characterization of commercial microemulsions (Gelucire® 44/14), used as an excipient in the pharmaceutical formulation, is provided with a comparison to DLS measurements. It was found that the polydispersity in size of the micelle did not influence the Gaussian peak shape of the taylorgram due to rapid surfactant exchange compared to the time-scale of the experiments (a few minutes).
Keywords: Taylor dispersion analysis; Hydrodynamic radius; Microemulsions; Micelles; Diffusion coefficient; Pharmaceutical excipient;

In vitro study of the specific interaction between poly(2-dimethylamino ethylmethacrylate) based polymers with platelets and red blood cells by Luca Flebus; François Lombart; Lucía Martinez-Jothar; Chantal Sevrin; Céline Delierneux; Cécile Oury; Christian Grandfils (55-64).
Display OmittedPoly(2-dimethylamino)ethyl methacrylate (PDMAEMA) is an attractive polycation frequently proposed as a non-viral vector for gene therapy. As expected for other cationic carriers, intravenous administration of PDMAEMA can result in its ionic complexation with various negatively charged domains found within the blood. To gain more insight into this polycation hemoreactivity, we followed the binding kinetics of a free form (FF) of fluorescein labelled PDMAEMA (Mn below 15 kDa) in normal human blood using flow cytometry. This in vitro study highlighted that platelets display higher affinity for this polycation compared to red blood cells (RBCs), with an adsorption isotherm characteristics of a specific saturable binding site. PDMAEMA (1–20 μg/mL) exerted a concentration dependent proaggregant effect with a biphasic aggregation of washed platelets. Activation of platelets was also noticed in whole blood with the expression of P-selectin and fibrinogen on platelet surface. Although additional studies would be needed in order to elucidate the mechanism of PDMAEMA mediated activation of platelets, our manuscript provides important information on the hemoreactivity of FF PDMAEMA.
Keywords: Poly(2-dimethylamino-ethylmethacrylate); Blood cells; Polycation; Hemocompatibility;

Optimization of nanostructured lipid carriers loaded with methotrexate: A tool for inflammatory and cancer therapy by Mara Ferreira; Luíse L. Chaves; Sofia A. Costa Lima; Salette Reis (65-72).
Display OmittedThe aim of this study was to optimize and assess the potential of nanostructured lipid carriers (NLC), prepared by the hot ultrasonication method, as carrier for methotrexate (MTX), highlighting the application of factorial design. Preliminary screening drug/lipid solubility, allowed us to select Witepsol® E85 as the solid lipid and Mygliol® 812 as liquid lipid for the NLC loaded with MTX. Then, a 3-level, 3-factor Box-Behnken design and validated by ANOVA analysis; the correspondence between the predicted values and those measured experimentally confirmed the robustness of the design. Properties of optimized MTX-loaded NLCs such as morphology, size, zeta potential, entrapment efficiency, storage stability, in vitro drug release and cytotoxicity were investigated. NLCs loaded with MTX exhibited spherical shape with 252-nm, a polydispersity of 0.06 ± 0.02, zeta potential of −14 mV and an entrapment efficiency of 87%. In vitro release studies revealed a fast initial release followed by a prolonged release of MTX from the NLC up to 24-h. The release kinetics of the optimized NLC best fitted the Peppas–Korsmeyer model for physiological and inflammatory environments and the Hixson–Crowell model skin simulation conditions. No toxicity was observed in fibroblasts. Thus, the optimized MTX-loaded NLC have the potential to be exploited as delivery system.
Keywords: Lipid colloidal carriers; Methotrexate; Witepsol® E85; Mygliol® 812; Box–Behnken design; Hot ultrasonication; In vitro drug release; Storage stability; L929 fibroblasts;

Design of controlled release systems for THEDES—Therapeutic deep eutectic solvents, using supercritical fluid technology by Ivo M. Aroso; Rita Craveiro; Ângelo Rocha; Madalena Dionísio; Susana Barreiros; Rui L. Reis; Alexandre Paiva; Ana Rita C. Duarte (73-79).
Display OmittedDeep eutectic solvents (DES) can be formed by bioactive compounds or pharmaceutical ingredients. A therapeutic DES (THEDES) based on ibuprofen, a non-steroidal anti-inflammatory drug (NSAID), and menthol was synthesized and its thermal behavior was analyzed by differential scanning calorimetry (DSC). A controlled drug delivery system was developed by impregnating a starch:poly-ϵ-caprolactone polymeric blend (SPCL 30:70) with the menthol:ibuprofen THEDES in different ratios (10 and 20 wt%), after supercritical fluid sintering at 20 MPa and 50 °C. The morphological characterization of SPCL matrices impregnated with THEDES was performed by scanning electron microscopy (SEM) and micro-computed tomography (micro-CT). Drug release studies were carried out in a phosphate buffered saline. The results obtained provide important clues for the development of carriers for the sustainable delivery of bioactive compounds.
Keywords: Therapeutic deep eutectic solvents; Supercritical carbon dioxide; Drug delivery systems; Biodegradable polymers; Ibuprofen;

Development and evaluation of folate functionalized albumin nanoparticles for targeted delivery of gemcitabine by Ravindra Dhar Dubey; Noor Alam; Ankit Saneja; Vaibhav Khare; Ashok Kumar; Shagun Vaidh; Girish Mahajan; Parduman R. Sharma; Shashank K. Singh; Dilip M. Mondhe; Prem N. Gupta (80-91).
Display OmittedGemcitabine is one of the most potent anticancer agents acting on a wide range of solid tumors, however, its use is limited by short half life and high dose leading to serious side effects. The present investigation describes the development and characterization of folate functionalized gemcitabine loaded bovine serum albumin nanoparticles (Fa-Gem-BSANPs). The nanoparticles were prepared by desolvation cross-linking technique and characterized for various parameters including morphology, particle size, zeta potential, drug loading and release profile. The particle size of Gem-BSANPs and Fa-Gem-BSANPs was found to be 159.1 ± 5.29 and 208.7 ± 1.80 nm, respectively. DSC and XRD analysis indicated amorphous nature of the drug within the particles. The encapsulated gemcitabine exhibited less hemolytic properties as compared to native drug. The anticancer activity of Fa-Gem-BSANPs was evaluated in folate receptor over expressing cell lines (Ovcar-5 and MCF-7) and folate receptor deficient cell line (MIAPaCa-2). The Fa-Gem-BSANPs showed superior anticancer activity as compared to Gem-BSANPs in Ovcar-5 and MCF-7 cells while no significant difference in cytotoxicity was found with MIAPaCa-2 cells. Confocal microscopy indicated facilitated intracellular uptake of Fa-Gem-BSANPs in MCF-7, which in turn result in a higher potential for apoptosis. Further, Fa-Gem-BSANPs exhibited improved anti-tumor activity in Ehrlich solid tumor model in mice. In conclusion, our study indicates that folate functionalized nanoparticles confer enhance cellular uptake and cytotoxicity for gemcitabine.
Keywords: Gemcitabine; Albumin; Chemotherapy; Nanoparticles; Active targeting;

Layer-by-layer engineered nanocapsules of curcumin with improved cell activity by Paveenuch Kittitheeranun; Warayuth Sajomsang; Sarunya Phanpee; Alongkot Treetong; Tuksadon Wutikhun; Kunat Suktham; Satit Puttipipatkhachorn; Uracha Rungsardthong Ruktanonchai (92-102).
Display OmittedNanocarriers based on electrostatic Layer-by-layer (LbL) assembly of CaCO3 nanoparticles (CaCO3 NPs) was investigated. These inorganic nanoparticles was used as templates to construct nanocapsules made from films based on two oppositely charged polyelectrolytes, poly(diallyldimethylammonium chloride), and poly (sodium 4-styrene-sulfonate sodium salt), followed by core dissolution. The naked CaCO3 NPs, CaCO3 NPs coated with the polyelectrolytes and hollow nanocapsules were found with hexagonal shape with average sizes of 350–400 nm. A reversal of the surface charge between positive to negative zeta potential values was found, confirming the adsorption of polyelectrolytes. The loading efficiency and release of curcumin were controlled by the hydrophobic interactions between the drug and the polyelectrolyte matrix of the hollow nanocapsules. The quantity of curcumin released from hollow nanocapsules was found to increase under acidic environments, which is a desirable for anti-cancer drug delivery. The hollow nanocapsules were found to localize in the cytoplasm and nucleus compartment of Hela cancer cells after 24 h of incubation. Hollow nanocapsules were non-toxic to human fibroblast cells. Furthermore, curcumin loaded hollow nanocapsules exhibited higher in vitro cell inhibition against Hela cells than that of free curcumin, suggesting that polyelectrolyte based-hollow nanocapsules can be utilized as new carriers for drug delivery.
Keywords: Layer-by-layer; Polyelectrolyte multilayer; Calcium carbonate; Curcumin; Hollow nanocapsules;

Display OmittedThe interaction of small biologically active molecules in the nanocavity of supramolecular host is very interesting and thriving research area. In the presence of supramolecular host the absorption and emission properties of small biologically active molecules were modulated several folds compared to bulk solution. In this study we have investigated the supramolecular interaction of a cancer cell photosensitizer molecule harmane in the presence of cucurbit[7]uril (CB7) as host in aqueous buffer solution (pH ∼ 7.2). We have used steady state absorption, emission and time resolved fluorescence spectroscopy techniques. The thermodynamics of the binding between harmane in the nanochannel of CB7 were studied by using isothermal titration calorimetry (ITC) method. The emission properties of harmane are modulated several fold in the presence of CB7. ITC study indicates that the complexation between harmane and CB7 are enthalpically favourable.
Keywords: Drug encapsulation; Cucurbit[7]uril; Cancer cell photosensitizer; Isothermal titration calorimetry; Fluorescence;

Male enhancement Nutraceuticals in the Middle East market: Claim, pharmaceutical quality and safety assessments by Ghada ElAgouri; Fatema ElAmrawy; Ahmed ElYazbi; Ahmed Eshra; Mohamed I. Nounou (109-119).
Display OmittedThe global market is invaded by male enhancement nutraceuticals claimed to be of natural origin sold with a major therapeutic claim. Most of these products have been reported by international systems like the Food and Drug Administration (FDA). We hypothesize that these products could represent a major threat to the health of the consumers. In this paper, pharmaceutical evaluation of some of these nutraceutical products sold in Egypt under the therapeutic claim of treating erectile dysfunction, are discussed along with pharmacological evaluation to investigate their safety and efficacy parameters. Samples were analyzed utterly using conventional methods, i.e.: HPLC, HPTLC, NIR, content uniformity and weight variation and friability. The SeDeM system was used for quality assessment. On the basis of the results of this research, the sampled products are adulterated and totally heterogeneous in their adulterant drug content and pharmaceutical quality. These products represent a major safety threat for the consumers in Egypt and the Middle East, especially; the target audience is mostly affected with heart and blood pressure problems seeking natural and safe alternatives to the well-established Phosphodiesterase 5 Inhibitors (PDE-5Is).
Keywords: Phosphodiestrase inhibitors; Sildenafil; Clinical Study; SeDeM; Evaluation; Adulteration; Middle East; Nutraceuticals; Tiger King;

Liposomes as a potential ocular delivery system of distamycin A by Patrizia Chetoni; Daniela Monti; Silvia Tampucci; Barbara Matteoli; Luca Ceccherini-Nelli; Alessando Subissi; Susi Burgalassi (120-126).
Display OmittedLiposomes containing Distamycin A (DA) may be clinically useful in the treatment of ocular HSV infections, especially in acyclovir-resistant HSV keratitis. This study evaluated the in vitro and in vivo performance of a topical controlled release liposomal formulation containing DA (DA-Lipo) aimed at reducing the toxicity of the encapsulated active agent and improving drug uptake by ocular tissues. The bioavailability of DA in the tear fluid and the DA uptake into the cornea were increased after instillation of DA-Lipo in rabbits, reaching the DA corneal concentration corresponding to IC50 values against HSV without any sign of transcorneal permeation of drug. DA-Lipo was definitely less cytotoxic then plain DA in rabbit corneal epithelial cells. These results provide new insights into the correlation between the in vitro data and the drug kinetics following ocular applications of liposomal vesicles.
Keywords: Liposome; Distamycin A; Antiviral; Pharmacokinetic; Cytotoxicity; Rabbit;

Evaluation of etanercept degradation under oxidative stress and potential protective effects of various amino acids by Jun Yeul Lim; Nam Ah Kim; Dae Gon Lim; Ki Hyun Kim; Shavron Hada; Seong Hoon Jeong (127-136).
Display OmittedTo evaluate the oxidative stability of proteins, a model protein, etanercept, was exposed to oxidative stress conditions using hydrogen peroxide. Various amino acids were also evaluated on their antioxidant effect. Transition temperature (T m), secondary structural content, hydrodynamic size, and aggregation and fragmentation of etanercept in solution were assessed using dynamic light scattering (DLS), size exclusion chromatography (SEC), differential scanning calorimetry (DSC), and far-UV circular dichroism (CD). Sample solutions were stored at 4 °C, 20 °C, and 40 °C under oxidative stress. The DLS results exhibited a decrease in the Z-average and intensity peak size of etanercept during the storage, suggesting fragmentation issues rather than aggregation by oxidation. The SEC results exhibited an increase in fragmentation and a decrease in aggregation and monomer content. The monomer content remained higher in histidine than in other amino acids, followed by methionine. There were three T m of etanercept that were selected as key parameters of conformational stability. Oxidized samples exhibited a significant decrease in T m values, indicating decreased conformational stability. Methionine exhibited the highest values in T m1, followed by histidine. The CD spectrum exhibited one unique negative peak of etanercept without amino acids, and changed with oxidation. Only methionine exhibited an enhancement of the stability. All four biophysical analyses results suggest that the histidine and methionine provide a protective effect in the protein solution against oxidative stress. However, histidine was effective as an antioxidant but methionine showed highly enhanced conformational and secondary structural stability.
Keywords: Etanercept; Oxidation; Amino acids; Methionine; Histidine; Biophysical methods;

Micafungin is a costly treatment and packaging of 50 mg or 100 mg bottles only are available, while doses lower than 5 mg and 20 mg are often necessary in neonates and paediatrics patients, respectively. The stability of micafungin sodium in polypropylene syringes and glass bottles was studied at different concentrations. Solutions of micafungin diluted with NaCl 0.9% were prepared in glass bottles (20 and 10 mg/mL) or syringes (1 and 0.5 mg/mL) and stored at 25 °C, 60% humidity (RH), in the dark (ICH conditions). Solutions were also exposed to heat (70 °C) or alkaline solution (NaOH) in order to force degradation. Samples were analysed at days 1, 5, 8 (for bottles) and also 15 (for syringes) after the preparation and assayed in triplicate. Stability was studied using a stability-indicating high-performance liquid chromatographic method. Syringes stored at 25 °C retained over 90% of their initial concentration over the study period. Temperature and alkaline conditions had significant effect on the stability of micafungin, leading to apparition of degradation products. Moreover, sub visible particles were in the specification of the European Pharmacopeia along 15 days. To conclude, micafungin diluted in NaCl 0.9% and stored in polypropylene syringes was chemically stable for at least 15 days at 25 °C in the dark.
Keywords: Micafungin; Polypropylene syringe; HPLC; Stability; CIVAS;

Systemic delivery of micelles loading with paclitaxel using N-succinyl-palmitoyl-chitosan decorated with cRGDyK peptide to inhibit non-small-cell lung cancer by Zhi-qiang Yuan; Ji-zhao Li; Yang Liu; Wei-liang Chen; Shu-di Yang; Chun-ge Zhang; Wen-jing Zhu; Xiao-feng Zhou; Chun Liu; Xue-nong Zhang (141-151).
Display OmittedThis study aimed to prepare efficient cRGDyK peptide-decorated micelles for the targeted therapy of non-small-cell lung cancer (NSCLC). An amphiphilic copolymer N-succinyl-palmitoyl-chitosan (SPCS) was synthesized and characterized. cRGDyK peptide is a ligand that can target tumors via specific binding integrin receptor overexpressed on tumor neovascularization and cells. cRGDyK-functionalized SPCS micelles loaded with paclitaxel (PTX/cRGDyK-SPCS) were prepared by film dispersion method and then characterized according to morphology, size, and zeta potential. PTX/cRGDyK-SPCS micelles presented pH-triggered drug release behavior under acidic conditions. The accumulation of micelles detected by laser confocal fluorescence microscopy and flow cytometry showed that cRGDyK-SPCS micelles were easily taken up by A549 cells marked with the luciferase gene (luc-A549). Meanwhile, co-localization of the micelles and lysosomes was recorded dynamically using a live cell station. MTT assays and cell apoptosis studies revealed that cell viability was significantly inhibited by PTX/cRGDyK-SPCS micelles. More importantly, in vivo animal studies showed that cRGDyK-SPCS micelles mainly accumulated in the orthotopic tumor site. PTX/cRGDyK-SPCS micelles exhibited better anti-tumor activity in subcutaneous and orthotopic lung tumors compared with PTX/SPCS micelles and Taxol®. These results suggested that PTX/cRGDyK-SPCS micelles had better cancer targeting capacity and superior anti-tumor efficacy. Thus, these micelles have great potential as novel carriers in delivering anti-tumor drugs.
Keywords: Keywords: Chitosan; cRGDyK; Micelles; luc-A549; Orthotopic lung tumor;

Characteristic of core materials in polymeric micelles effect on their micellar properties studied by experimental and dpd simulation methods by Furong Cheng; Xuewa Guan; Huan Cao; Ting Su; Jun Cao; Yuanwei Chen; Mengtan Cai; Bin He; Zhongwei Gu; Xianglin Luo (152-160).
Display OmittedPolymeric micelles are one important class of nanoparticles for anticancer drug delivery, but the impact of hydrophobic segments on drug encapsulation and release is unclear, which deters the rationalization of drug encapsulation into polymeric micelles. This paper focused on studying the correlation between the characteristics of hydrophobic segments and encapsulation of structurally different drugs (DOX and β-carotene). Poly(ϵ-caprolactone) (PCL) or poly(l-lactide) (PLLA) were used as hydrophobic segments to synthesize micelle-forming amphiphilic block copolymers with the hydrophilic methoxy-poly(ethylene glycol) (mPEG). Both blank and drug loaded micelles were spherical in shape with sizes lower than 50 nm. PCL-based micelles exhibited higher drug loading capacity than their PLLA-based counterparts. Higher encapsulation efficiency of β-carotene was achieved compared with DOX. In addition, both doxorubicin and β-carotene were released much faster from PCL-based polymeric micelles. Dissipative particle dynamics (DPD) simulation revealed that the two drugs tended to aggregate in the core of the PCL-based micelles but disperse in the core of PLLA based micelles. In vitro cytotoxicity investigation of DOX loaded micelles demonstrated that a faster drug release warranted a more efficient cancer-killing effect. This research could serve as a guideline for the rational design of polymeric micelles for drug delivery.
Keywords: Dissipative particle dynamics simulation; Encapsulation; Polymeric micelles; Chain characteristic; Drug delivery;

Display OmittedRecently there has been a growth of interest in the novel skin care formulations containing active ingredients such as low molecular weight peptides. In this paper we present new skincare formulations such as hydrogels, oil-in-water emulsions and water-in-oil emulsion containing a tetrapeptide (N-acetyl-Pro-Pro-Tyr-Leu). These formulations were characterized in terms of physicochemical parameters (pH, viscosity), stability and particle size distribution. Additionally, the diffusion parameters of the peptide in the obtained formulations were calculated based on the Einstein–Smoluchowski equation. Furthermore, in order to determine the penetration of the tetrapeptide through membranes its release kinetics were investigated. On the basis of release curves, the release rate constants were determined. The results proved that the properties of the formulations strongly determined the release rate of the tetrapeptide. The higher viscosity of the semisolid, the slower was the permeation through the membrane.
Keywords: Low molecular peptides; Diffusion test; Topical formulations; Release rate; Einstein–Smoluchowski equation;

The potential of adjuvants to improve immune responses against TdaP vaccines: A preclinical evaluation of MF59 and monophosphoryl lipid A by Valentina Agnolon; Cristina Bruno; Rosanna Leuzzi; Bruno Galletti; Ugo D’Oro; Mariagrazia Pizza; Anja Seubert; Derek T. O’Hagan; Barbara C. Baudner (169-176).
Display OmittedThe successful approach of combining diphtheria, tetanus and pertussis antigens into a single vaccine has become a cornerstone of immunization programs. Yet, even if vaccination coverage is high, a resurgence of pertussis has been reported in many countries suggesting current vaccines may not provide adequate protection. To induce better tailored and more durable immune responses against pertussis vaccines different approaches have been proposed, including the use of novel adjuvants. Licensed aP vaccines contain aluminum salts, which mainly stimulate humoral immune responses and might not be ideal for protecting against Bordetella pertussis infection. Adjuvants inducing more balanced T-helper profiles or even Th1-prone responses might be more adequate. In this study, two adjuvants already approved for human use have been tested: MF59 emulsion and the combination of aluminum hydroxide with the Toll-Like Receptor 4 agonist MPLA. Adjuvanticity was evaluated in a mouse model using a TdaP vaccine containing three B. pertussis antigens: genetically detoxified pertussis toxin (PT-9K/129G), filamentous hemagglutinin (FHA) and pertactin (PRN)The physico-chemical compatibility of TdaP antigens with the proposed adjuvants, together with a quicker onset and changed quality of the antibody responses, fully supports the replacement of aluminum salts with a new adjuvant to enhance aP vaccines immunogenicity.
Keywords: Adjuvants; Vaccine formulation; Pertussis; Mouse model;

UV-curable gel formulations: Potential drug carriers for the topical treatment of nail diseases by Laxmi Valji Kerai; Stephen Hilton; Sudaxshina Murdan (177-190).
Display OmittedNail diseases are common, cause significant distress and treatments are far from successful. Our aim was to investigate the potential of UV-curable gels – currently used as cosmetics – as topical drug carriers for their treatment. These formulations have a long residence on the nail, which is expected to increase patient compliance and the success of topical therapy. The gels are composed of the diurethane dimethacrylate, ethyl methacrylate, 2-hydroxy-2-methylpropiophenone, an antifungal drug (amorolfine HCl or terbinafine HCl) and an organic liquid (ethanol or NMP) as drug solvent. Following its application to a substrate and exposure to a UVA lamp for 2 min, the gel polymerises and forms a smooth, glossy and amorphous film, with negligible levels of residual monomers. No drug-polymer interactions were found and drug loading did not affect the film’s properties, such as thickness, crystallinity and transition temperatures. In contrast, the organic solvent did influence the film’s properties; NMP-containing films had lower glass transition temperatures, adhesion and water resistance than ethanol-based ones. Water-resistance being a desired property, ethanol-based formulations were investigated further for stability, drug release and ungual permeation. The films were stable under accelerated stability testing conditions. Compared to terbinafine, amorolfine was released to a greater extent, had a higher ungual flux, but a lower concentration in the nailplate. However, both drugs were present at considerably high levels in the nail when their MICs are taken into account. We thus conclude that UV-curable gels are promising candidates as topical nail medicines.
Keywords: UV gel; Acrylates; Ungual; Release; Permeation; Onychomycosis;

Facile synthesis of functionalized ionic surfactant templated mesoporous silica for incorporation of poorly water-soluble drug by Jing Li; Lu Xu; Baixue Yang; Hongyu Wang; Zhihong Bao; Weisan Pan; Sanming Li (191-198).
Display OmittedThe present paper reported amino group functionalized anionic surfactant templated mesoporous silica (Amino-AMS) for loading and release of poorly water-soluble drug indomethacin (IMC) and carboxyl group functionalized cationic surfactant templated mesoporous silica (Carboxyl-CMS) for loading and release of poorly water-soluble drug famotidine (FMT). Herein, Amino-AMS and Carboxyl-CMS were facilely synthesized using co-condensation method through two types of silane coupling agent. Amino-AMS was spherical nanoparticles, and Carboxyl-CMS was well-formed spherical nanosphere with a thin layer presented at the edge. Drug loading capacity was obviously enhanced when using Amino-AMS and Carboxyl-CMS as drug carriers due to the stronger hydrogen bonding force formed between surface modified carrier and drug. Amino-AMS and Carboxyl-CMS had the ability to transform crystalline state of loaded drug from crystalline phase to amorphous phase. Therefore, IMC loaded Amino-AMS presented obviously faster release than IMC because amorphous phase of IMC favored its dissolution. The application of asymmetric membrane capsule delayed FMT release significantly, and Carboxyl-CMS favored sustained release of FMT due to its long mesoporous channels and strong interaction formed between its carboxyl group and amino group of FMT.
Keywords: Mesoporous silica; Ionic surfactant template; Amine modification; Carboxylic modification; Drug incorporation;

Near-infrared spectroscopy for the detection and quantification of bacterial contaminations in pharmaceutical products by Cristina Quintelas; Daniela P. Mesquita; João A. Lopes; Eugénio C. Ferreira; Clara Sousa (199-206).
Display OmittedAccurate detection and quantification of microbiological contaminations remains an issue mainly due the lack of rapid and precise analytical techniques. Standard methods are expensive and time-consuming being associated to high economic losses and public health threats. In the context of pharmaceutical industry, the development of fast analytical techniques able to overcome these limitations is crucial and spectroscopic techniques might constitute a reliable alternative. In this work we proved the ability of Fourier transform near infrared spectroscopy (FT-NIRS) to detect and quantify bacteria (Bacillus subtilis, Escherichia coli, Pseudomonas fluorescens, Salmonella enterica, Staphylococcus epidermidis) from 10 to 108  CFUs/mL in sterile saline solutions (NaCl 0.9%). Partial least squares discriminant analysis (PLSDA) models showed that FT-NIRS was able to discriminate between sterile and contaminated solutions for all bacteria as well as to identify the contaminant bacteria. Partial least squares (PLS) models allowed bacterial quantification with limits of detection ranging from 5.1 to 9 CFU/mL for E. coli and B. subtilis, respectively. This methodology was successfully validated in three pharmaceutical preparations (contact lens solution, cough syrup and topic anti-inflammatory solution) proving that this technique possess a high potential to be routinely used for the detection and quantification of bacterial contaminations.
Keywords: FT-NIRS; Bacterial contaminations; Pharmaceutical products; Chemometrics; Process analytical technology;

Use of anodized titanium alloy as drug carrier: Ibuprofen as model of drug releasing by Antonio L. Doadrio; A. Conde; M.A. Arenas; J.M. Hernández-López; J.J. de Damborenea; Concepción Pérez-Jorge; Jaime Esteban; Maria Vallet-Regí (207-212).
Display OmittedThe use of osteoarticular implants has improved the quality of life of millions of patients. In this work nanotubular structures tailored made on Ti6Al4 V substrates was used as drug delivery system of ibuprofen as a proof of concept. Three different nanotubular films with different sizes and forms (NT, NT+ and NTb) were analysed. Samples were soaked in a solution of 660 mg ibuprofen/20 mL n-pentane. The ibuprofen release in aqueous medium was evaluated by liquid chromatography reversed-phase (RP-HPLC). To calculate the observed constant k, the amount of ibuprofen released was plotted versus the time using linear regression according to the zero-order, first-order, second-order and Higuchi model. The release of ibuprofen was constant and independent of the concentration. The kinetic constant obtained was 0.021 (NT), 0.022 (NT+) and 0.013 (NTb) being the correlation factor of 0.98 (zero-order) where the maximum correlation factor was reached. These results indicate that the delivery process from NT and NT+ is similar and slower that NTb. In all the cases was inside the therapeutically range. These results showed the potential of these modifications in order to develop implants that can carry different molecules of medical importance.
Keywords: Anodization; Ti6Al4V; Ibuprofen; Releasing; Nanotubes;

Dry powder pulmonary delivery of cationic PGA-co-PDL nanoparticles with surface adsorbed model protein by Nitesh K. Kunda; Iman M. Alfagih; Sarah R. Dennison; Satyanarayana Somavarapu; Zahra Merchant; Gillian A. Hutcheon; Imran Y. Saleem (213-222).
Display OmittedPulmonary delivery of macromolecules has been the focus of attention as an alternate route of delivery with benefits such as; large surface area, thin alveolar epithelium, rapid absorption and extensive vasculature. In this study, a model protein, bovine serum albumin (BSA) was adsorbed onto cationic PGA-co-PDL polymeric nanoparticles (NPs) prepared by a single emulsion solvent evaporation method using a cationic surfactant didodecyldimethylammonium bromide (DMAB) at 2% w/w (particle size: 128.64 ± 06.01 nm and zeta-potential: +42.32 ± 02.70 mV). The optimum cationic NPs were then surface adsorbed with BSA, NP:BSA (100:4) ratio yielded 10.01 ± 1.19 μg of BSA per mg of NPs. The BSA adsorbed NPs (5 mg/ml) were then spray-dried in an aqueous suspension of L-leucine (7.5 mg/ml, corresponding to a ratio of 1:1.5/NP:l-leu) using a Büchi-290 mini-spray dryer to produce nanocomposite microparticles (NCMPs) containing cationic NPs. The aerosol properties showed a fine particle fraction (FPF, dae < 4.46 μm) of 70.67 ± 4.07% and mass median aerodynamic diameter (MMAD) of 2.80 ± 0.21 μm suggesting a deposition in the respiratory bronchiolar region of the lungs.The cell viability was 75.76 ± 03.55% (A549 cell line) at 156.25 μg/ml concentration after 24 h exposure. SDS-PAGE and circular dichroism (CD) confirmed that the primary and secondary structure of the released BSA was maintained. Moreover, the released BSA showed 78.76 ± 1.54% relative esterolytic activity compared to standard BSA.
Keywords: Dry powder inhalation; Nanoparticles; Pulmonary delivery; Spray drying; Proteins;

Development of antimigraine transdermal delivery systems of pizotifen malate by C.E. Serna-Jiménez; S. del Rio-Sancho; M.A. Calatayud-Pascual; C. Balaguer-Fernández; A. Femenía-Font; A. López-Castellano; V. Merino (223-232).
Display OmittedThe aim of this study was to develop and evaluate a transdermal delivery system of pizotifen malate. Pizotifen is frequently used in the preventive treatment of migraine, but is also indicated in eating disorders.In the course of the project, the effects of chemical enhancers such as ethanol, 1,8-cineole, limonene, azone and different fatty acids (decanoic, decenoic, dodecanoic, linoleic and oleic acids) were determined, first using a pizotifen solution. Steady state flux, diffusion and partition parameters were estimated by fitting the Scheuplein equation to the data obtained. Among the chemical enhancers studied, decenoic acid showed the highest enhancement activity, which seemed to be due to the length of its alkyl chain and unsaturation at the 9th carbon. The influence of iontophoresis and the involvement of electrotransport in said process was determined. The absorption profile obtained with iontophoresis was similar to that obtained with fatty acids and terpenes, though skin deposition of the drug was lower with the former. Transdermal delivery systems (TDS) of pizotifen were manufactured by including chemical enhancers, decenoic acid or oleic acid, and were subsequently characterized. When the results obtained with solutions were compared with those obtained with the TDS, a positive enhancement effect was observed with the latter with respect to the partitioning and diffusion of the drug across the skin. Our findings endorse the suitability of our TDS for delivering therapeutic amounts of pizotifen malate.
Keywords: Pizotifen; Transdermal; TDS; Chemical enhancers; Iontophoresis; Migraine;

Display OmittedRecently, non-viral vectors have become a popular research topic in the field of gene therapy. In this study, we conjugated short oligopeptides to polyamidoamine-generation 4 (PAMAM G4) to achieve higher transfection efficiency. Previous reports have shown that the PAMAM G4-histidine (H)-arginine (R) dendrimer enhances gene delivery by improving cell penetration and internalization mechanisms. Therefore, we synthesized PAMAM G4-H phenylalanine (F) R, PAMAM G4-FHR and PAMAM G4-FR derivatives to determine the best gene carrier with the lowest toxicity. Physicochemical studies were performed to determine mean diameters and surface charge of PAMAM derivatives/pDNA polyplexes. DNA condensation was confirmed using a gel retardation assay. Cytotoxicity and transfection efficiency were analyzed using human cervical carcinoma (HeLa) and human liver carcinoma (HepG2) cells. Similar levels of transfection were achieved in both cell lines by using gold standard transfection reagent PEI 25 kD. Therefore, our results show that these carriers are promising and may help achieve higher transfection with negligible cytotoxicity.
Keywords: Polyamidoamine dendrimer; Oligopeptides; Hydrophobic; Transfection; Polyplex; Cytotoxicity;

Display OmittedNovel drug delivery systems have previously never been used in the formulation of any crude unfractionated traditional Chinese medicine. In the present study, a multi-unit drug delivery system (MUDDS) for a Chinese medicine Niuhuang Xingxiao Wan (NXW) was prepared using micro/nanotechnologies to enhance the bioavailability and efficacy. NXW was formulated into four units, that is, realgar, frankincense and myrrh oil (FMO), musk, and bezoar. The four units were processed using the wet ball milling, high-pressure homogenization, liquid nitrogen freezing-ultracentrifugation trituration, and dry grinding methods, respectively. After formulation, the four independent units prepared were encapsulated together to obtain the final NXW–MUDDS. Pharmacokinetic studies showed that the area under the plasma concentration–time curve (AUC), terminal half-life (T 1/2), and time to reach the peak plasma concentration (T max) following administration of NXW–MUDDS were 5.21, 1.96, and 1.99 times higher, respectively, than that of NXW. The in vivo antitumor activity assay showed that the efficacy of NXW–MUDDS was significantly higher (P  < 0.05) than that of NXW. Collectively, these results reveal the feasibility of applying micro/nanotechnologies in formulating Chinese medicines.
Keywords: Niuhuang Xingxiao Wan; Micro/nanotechnology; Chinese medicinal formula; Multi-unit drug delivery system;

Lyophilized sponges loaded with curcumin solid lipid nanoparticles for buccal delivery: Development and characterization by Heba A. Hazzah; Ragwa M. Farid; Maha M.A. Nasra; Magda A. EL-Massik; Ossama Y. Abdallah (248-257).
Display OmittedThis study aimed to prepare and evaluate mucoadhesive sponges as dosage forms for delivering solid lipid nanoparticles. For this purpose curcumin (Cur) was formulated as solid nanoparticles (SLN) using Gelucire 50/13, and polaxomer 407. The prepared CurSLN dispersion was thickened with different mucoadhesive polymers. Different concentrations of glycerol, and mannitol of range (0.25–20%), and (0–1%), respectively were also examined. The formed gel was poured into oblong molds and freeze dried to form mucoadhesive sponge to be applied to the buccal mucosa. The prepared sponges were evaluated for their, in-vivo residence time, in-vitro and in-vivo drug release, and hydration capacity. Surface morphology for the different sponges were examined using SEM. TEM was also carried out for sponge fragments previously dispersed into water. Infrared spectroscopy was conducted to investigate interaction between used ingredients. The results showed that the CurSLN loaded HPMC, and Polycarbophil sponges showed 4, and 15 h in-vivo residence time, respectively, providing a considerable amount of curcumin into saliva. The incorporation of glycerol and mannitol at concentration of 1% provided elegant and flexible sponges. The SEM showed that the deposition of CurSLN differed according to the type of polymer used. TEM confirmed the integrity of liberated CurSLN from sponges. IR spectra showed an interaction between HPMC and poloxamer 407, which affected its behavior as a gelling agent. The obtained results provide an efficient approach for delivering solid lipid nanoparticles in a solid dosage form keeping the nanoparticle characters and integrity.
Keywords: Curcumin; Mucoadhesive; Sponge; Solid lipid nanoparticles;

Display OmittedHPMC capsules are made by a dipping process and a surface lubricant for the mould pins is an essential processing aid for removing dried capsules shells. For the purpose of this study, the level was determined by quantifying methyloleate (MO) a component found in the lubricant but not in the hypromellose capsules. Here we investigated the influence of the lubricant, low (10.81 μg/capsule = 60 mg/kg MO), medium (15.97 μg/capsule = 90 mg/kg MO) and high (23.23 μg/capsule = 127 mg/kg MO) content on powder (binary mixture of salbutamol: lactose, 1:50 w/w) aerosolization properties was investigated. Results indicated significantly lower emitted dose from capsules with 60 mg/kg MO. Furthermore, the 90 and 127 mg/kg MO level of lubricant capsules produced almost double the Fine Particle Dose & Fine Particle Fraction compared with the low level of lubricant. The data indicates that lubricant level within capsules has an influence on deposition profiles and amount of drug remaining in capsule and inhaler device after actuation. It is suggested lubricant levels greater than 60 mg/kg MO per capsule are required to minimise powder retention within capsules and maximise deposition profiles. AFM (atomic force microscopy) data suggest that internal surface roughness may be related with this phenomena.
Keywords: Hypromellose (HPMC) capsules; Dry powder inhalation; Lubricant; Inhaler; Atomic force microscopy (AFM);

Cited but not read and read but not cited by Alexander T. Florence (264-265).

Display OmittedDendrimers due to their unique architecture may play an important role in drug delivery systems including chemotherapy, gene therapy and recently, photodynamic therapy as well. We investigated two dendrimer-photosensitizer systems in context of potential use of these systems in photodynamic therapy. The mixtures of an anionic phosphorus dendrimer of the second generation and methylene blue were studied by UV–vis spectroscopy while that of a cationic phosphorus dendrimer (third generation) and rose bengal were investigated by spectrofluorimetric methods. Spectroscopic analysis of these two systems revealed the formation of dendrimer-photosensitizer complexes via electrostatic interactions as well as π stacking. The stoichiometry of the rose bengal-cationic dendrimer complex was estimated to be 7:1 and 9:1 for the methylene blue-anionic dendrimer complex. The results suggest that these polyanionic or polycationic phosphorus dendrimers can be promising candidates as carriers in photodynamic therapy.
Keywords: Phosphorus dendrimer; Photosensitizer; Rose bengal; Methylene blue; Photodynamic therapy; Drug delivery system;

Display OmittedThe review presents a full library of single-isomer primary rim per-carboxylate- and per-sulfate-α-, -β- and -γ-cyclodextrin (CD) derivatives and their potential for pharmaceutical nanotechnology. Recent advances in cyclodextrin chemistry have enabled robust methods for the synthesis of single-isomer anionic CDs. Numerous nanobio-applications have been already reported for these negatively charged derivatives, which alone or in combination with other biodegradable molecular platforms can become important carriers for targeted drug delivery and release. Specialized applications are also discussed, such as chiral separations, as well as the ability of per-6-carboxylated-cyclodextrins to coordinate with metal cations and especially with lanthanide cations that makes them candidates as contrast agents for Magnetic Resonance Imaging.
Keywords: Anionic cyclodextrins; Negatively charged; Carboxylate-; Sulfate-; Drug carriers; Cation binding;

Lipid-based nanoparticles containing cationic derivatives of PTA (1,3,5-triaza-7-phosphaadamantane) as innovative vehicle for Pt complexes: Production, characterization and in vitro studies by Rita Cortesi; Chiara Damiani; Laura Ravani; Lorenza Marvelli; Elisabetta Esposito; Markus Drechsler; Antonella Pagnoni; Paolo Mariani; Fabio Sforza; Paola Bergamini (291-300).
Display OmittedThe aliphatic phosphine PTA (1,3,5-triaza-7-phosphaadamantane) is a promising ligand for metal complexes designed and developed as innovative inorganic drugs.In the present paper, an N-alkylated derivative of PTA, (PTAC16H33)X (X = I, C1, or X = PF6, C2) and its platinum coordination complex cis-[PtCl2(PTAC16H33)2](PF6)2, C3, were considered as components of cationic lipid nanoparticles (CLNs). Particularly, CLN1, CLN2 and CLN3 were obtained by adding derivatives C1, C2 or C3 during nanoparticles preparation, while CLN2-Pt were obtained by treating preformed CLN2 with Pt(II). It was demonstrated that CLN1, CLN2 and CLN3 can be produced with technological conventional methods. However, among the two here proposed protocols, the one based on the treatment of preformed nanoparticles appears more advantageous as compared to the other since it allows a quantitative association yield of Pt. As determined by ICP-OES, a content of P and Pt 2.2-fold and 2.5-fold higher in CLN2-Pt than in CLN3 was evidenced. For the first time was demonstrated that properly functionalized preformed nanoparticles can be efficiently used to obtain a post production Pt(II) complex while maintaining a cytotoxic activity toward cultured cells. In fact, the antiproliferative activity shown by CLN3, CLN2-Pt on the three model cancer cell lines was substantially similar and comparable to that of complex C3 in dmso solution. Thus N-alkylated-PTA derivatives in CLNs could be proposed as innovative biocompatible and water dispersible nanoparticles carrying lipophilic Pt complexes becoming an interesting and improved system with respect to dmso solution.
Keywords: PTA; Cationic lipid nanoparticles; Cisplatin; Antiproliferative activity; Pt complexes;

Formulating better medicines for children—reflections by Hannah Batchelor; Smita Salunke; Catherine Tuleu (301-303).

Devices for oral and respiratory paediatric medicines: What do healthcare professionals think? by Jennifer Walsh; Marie-Christine Math; Jörg Breitkreutz; Thomas Zerback; Herbert Wachtel (304-315).
Display OmittedMedical devices are crucial for the proper administration of paediatric medicines to children, but handling and dosing errors commonly appear in daily practice. As both the understanding and the usage of medical devices for oral and respiratory drug administration are heterogeneous among patients and caregivers, the European Paediatric Formulation Initiative (EuPFI) consortium performed a European survey among healthcare professional stakeholders in France, Germany, Hungary, Italy, Spain and UK. The results show country- and age-dependent usage of devices for oral administration of liquid formulations, with a clear preference for oral droppers and syringes in the neonatal phase and in early infancy. In older children, spoons and cups are more frequently used although it is recognized that they may fail in delivering correct doses. The percentage of medicinal products definitely requiring an oral administration device was estimated as 68.8% by the participants. The survey elaborated a similar usage pattern for medical devices for respiratory drug delivery: in young children drug solutions are nebulized, using face-masks and subsequently valved holding chambers or spacers, with increasing age metered-dose inhalers and later dry powder inhalers are preferably used. 56% of the responding healthcare professionals believed that providing an administration device helps to ensure that the patient receives the correct dose of medicine, and 41% agreed that patients must be given an administration device with each supply of medicine. Interestingly, 6.7% thought that patients tend not to use the device provided and remarkably 25.4% stated that patients already have a device. Although there is the highest count of treated children with device supply in Germany and Hungary, there are no observed significant differences in the six investigated European countries (p  = 0.057). Patient difficulties in correct oral and respiratory device use were identified by respondents and potential solutions discussed.
Keywords: Paediatric; Respiratory device; Oral administration; Dosing device; Survey;

The STEP database through the end-users eyes—USABILITY STUDY by Smita Salunke; Catherine Tuleu (316-331).
Display OmittedThe user-designed database of Safety and Toxicity of Excipients for Paediatrics (“STEP”) is created to address the shared need of drug development community to access the relevant information of excipients effortlessly. Usability testing was performed to validate if the database satisfies the need of the end-users.Evaluation framework was developed to assess the usability. The participants performed scenario based tasks and provided feedback and post-session usability ratings. Failure Mode Effect Analysis (FMEA) was performed to prioritize the problems and improvements to the STEP database design and functionalities.The study revealed several design vulnerabilities. Tasks such as limiting the results, running complex queries, location of data and registering to access the database were challenging. The three critical attributes identified to have impact on the usability of the STEP database included (1) content and presentation (2) the navigation and search features (3) potential end-users.Evaluation framework proved to be an effective method for evaluating database effectiveness and user satisfaction. This study provides strong initial support for the usability of the STEP database. Recommendations would be incorporated into the refinement of the database to improve its usability and increase user participation towards the advancement of the database.
Keywords: Excipients; STEP database; Safety; Toxicity; Adverse effects; Paediatrics; Children; Neonates; Additives; Information resource;

Developmental pharmacology: A moving target by Janko Samardzic; Karel Allegaert; Milica Bajcetic (335-337).
Display OmittedThe main characteristic of pediatric and neonatal pharmacotherapy still is the insufficient availability of drugs with confirmed efficacy and safety data in children. Children differ from adults in the physiological, psychological and developmental terms and this subsequently results in differences in anticipated drug potency, efficacy and toxicity. This paper is focused on the most prominent issues of the contemporary developmental pharmacology.Child’s age and development can significantly affect drug pharmacokinetics (PK) processes. The dosage of drugs for children must be based on the physiological characteristics, as well as PK parameters of the drug obtained from the clinical trials with children. While knowledge about the impact of developmental changes on drug PK is increasing, information regarding pharmacodynamics (PD) is still more limited. The examples from clinical and animal data on ontogeny of receptors resulted in strong evidence for changes in drug response during development, in addition to but independent from PK alterations.In order to improve the use of medicines in children, it is essentially to know the complex processes of growth and development into the pediatric drug development programs. This is because absence of PK/PD data leads to increased risk of over- or under-dosing, adverse reactions or inefficiency.
Keywords: Children; Drugs; Developmental pharmacology; GABA;

Display OmittedSince the advent of the EU Paediatric Regulation in 2007, 78 of the 1688 Paediatric Investigation Plans (PIPs) have been for monoclonal antibodies (Mabs). Of these, 22 have been assessed by the MHRA. The purpose of this mini-review is to aid those researching and developing this class of drugs to better understand regulatory concerns leading to improved medicinal products for children. Three principal quality issues were identified for PIPs under Article 7 and 8: i) the level of anti-aggregation stabilisers, ii) acceptability and tolerability of administration (i.e. multiple injections, infusion time and volume), and iii) the need to develop new presentational forms (e.g. pre-filled syringe). Overall, two types of concerns were ascertained – those which are potentially avoidable (e.g. through development of new presentational forms) and others which require the evolution of new technologies in the sector (e.g. production of concentrated, stabilised preparations).
Keywords: Paediatric investigation plan; Monoclonal antibodies; Regulation; Children; MHRA; Injectable;

Formulation factors affecting acceptability of oral medicines in children by Fang Liu; Sejal Ranmal; Hannah K. Batchelor; Mine Orlu-Gul; Terry B. Ernest; Iwan W. Thomas; Talia Flanagan; Richard Kendall; Catherine Tuleu (341-343).
Display OmittedAcceptability of medicines in children and caregivers affects safety and effectiveness of medicinal treatments. The pharmaceutical industry is required to demonstrate acceptability of new paediatric formulations in target age groups as an integrated part of the development of these products (Kozarewicz, 2014). Two questions arise when trying to tackle this task: “which dosage form to choose for each target age group?” and “how to formulate it once the dosage form is decided?”. Inevitably, both the regulator and the developer turn to scientific evidence for answers. Research has emerged in recent years to demonstrate age-appropriateness and patient acceptability of different dosage forms; however, such information is still fragmented and far from satisfactory to define efficient formulation development strategies for a diverse patient subset (Ranmal and Tuleu, 2013). This paper highlights how formulation factors affect the acceptability of different oral medicines in children (Table 1), and it is based on a more extensive review article by Liu et al. (Liu et al., 2014). Gaps in knowledge are highlighted in order to stimulate further research. In some areas, findings from studies conducted in adult populations may provide useful guidance for paediatric development and this is also discussed.

Display OmittedIn the case of pediatric medicinal products the selection of an appropriate and palatable liquid dosage form can make the difference between treatment success and failure. Since the recent adoption of Pediatric Regulations in the U.S. and E.U., there is a greater demand for age-appropriate medicines for children. Extended research on the use of milk on drug administration in pediatric population has shown the multiple benefits of its use. Milk exhibits great solubilizing, gastroprotective and taste masking properties, which are very important characteristics in the case of insoluble, irritating and bitter-tasting active compounds. Milk-based formulations rely on a novel, simple and user-friendly approach for the delivery of ionized and unionized lipophilic drugs. In parallel they can provide critical nutritive elements and a wide range of biologically active peptides, very important elements especially for pediatric patients.
Keywords: Milk; Pediatric; Milk-based formulations; Regulatory;