European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences

Publications in this journal

• Article: Statins modulate feedback regulation mechanisms between advanced glycation end-products and C-reactive protein: evidence in patients with acute myocardial infarction.

  Beatriz Paradela-Dobarro, Sergio Raposeiras-Roubín, Bruno K Rodiño-Janeiro, Lilian Grigorian-Shamagian, José M García-Acuña, Pablo Aguiar-Souto, Michel Jacquet-Hervet, María V Reino-Maceiras, José R González-Juanatey, Ezequiel Alvarez
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  ABSTRACT: OBJECTIVE: High sensitivity C-reactive protein (hsCRP) and advanced glycation end-products (AGE) have been proposed as mediators in inflammation and atherosclerosis. Therefore, we studied the relation between AGE and hsCRP in patients with acute myocardial infarction (AMI). METHODS: Patients with AMI diagnosis and satisfying our inclusion criteria were included during 2009-2011 in an unicentre registry of AMI patients for a cross-sectional study. The final cohort was composed of 156 patients (46.2% STEMI and 27.6% with type-2 diabetes). AGE and hsCRP were measured in plasma. RESULTS: Diabetic patients were older than non-diabetics (68.6 ± 10.6 vs. 60.4 ± 13.9 years; p < 0.05), presented more incidence of hypertension (62.8 vs. 36.3%; p < 0.05) and were in a higher Killip class (p < 0.05). The mean values of fluorescent AGE and hsCRP levels were 61.3 ± 49.8 AU and 2.4 ± 4.0 mg/L, respectively, and there were no differences in these parameters between diabetic and non-diabetic patients. A direct association between AGE and hsCRP levels was observed, mainly in diabetic patients (r = 0.258; p = 0.018). Importantly, this association disappeared in patients who had been treated with statins before their AMI (r = -0.055; p = 0.845), but it was maintained in non-diabetic patients naïve for statins treatment (r = 0.634; p < 0.001), independently of other treatments and confounding parameters. CONCLUSIONS: This is the first evidence in humans of a feedback regulation mechanism between CRP and the AGE-RAGE axis modulated by statins.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: A Novel Manganese Complex, Mn-(II) N-(2-Hydroxy Acetophenone) Glycinate overcomes Multidrug-Resistance in Cancer.

  Ruma Dey Ghosh, Kaushik Banerjee, Satyajit Das, Avishek Ganguly, Paramita Chakraborty, Avijit Sarkar, Mitali Chatterjee, Soumitra K Choudhuri
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  ABSTRACT: Multidrug resistance (MDR) remains a significant problem for effective cancer chemotherapy. In spite of considerable advances in drug discovery, most of the cancer cases still stay incurable because of resistance to chemotherapy. We synthesized a novel, Mn (II) complex (chelate), viz., Manganese N-(2-hydroxy acetophenone) glycinate (MnNG) that exhibits considerable efficacy to overcome drug resistant cancer. The anti-proliferative activity of MnNG was studied on doxorubicin resistant and sensitive human T lymphoblastic leukemia cells (CEM/ADR 5000 and CCRF/CEM). MnNG induced apoptosis significantly in CEM/ADR 5000 cells probably through generation of reactive oxygen species. Moreover, intraperitoneal (i.p.) application of MnNG at nontoxic doses caused significant increase in the life-span of Swiss albino mice bearing sensitive and doxorubicin resistant subline of Ehrlich ascites carcinoma cells.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Overcoming therapeutic obstacles in inflammatory bowel diseases; A comprehensive review on novel drug delivery strategies.

  F Talaie, F Atyabi, M Azjdarzadeh, R Dinarvand, A Saadatzadeh
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  ABSTRACT: Inflammatory bowel diseases (IBD) are a group of debilitating inflammatory complications specially inflicting colonic tissue in which full long term remission with current standardized treatments is yet intangible. Therapeutic side effects and efficacy considerations necessitate the development of more effective systems which lower the required drug doses, reduce systemic adverse effects and deliver the drug specifically to the desired site of action in colon. The large surface area in large intestine is suitable for drug absorption but the primary approaches to treatment depend on the gastrointestinal (GI) condition and its movements. Hereafter, novel GI-independent targeted drug delivery systems including micro- and nanoparticles have been developed. Specially, owing to its safety and efficacy, biological therapy with antibodies which has recently been known as an alternative approach in maintenance of remission in IBD patients will be discussed. Furthermore, nano- and micro-therapeutic approaches to IBD treatment which have the potential to overcome some of the current drawbacks of conventional IBD therapy will be analyzed. This review provides broad but concise information over the arena of the evolving systems aimed at different targets involved in IBD which are being studied in animal or in vitro models of this complication, while comparing these to conventional treatments. It further discusses important pros and cons of therapeutic approaches against IBD while helping to better understand and evaluate the future impact of novel drug delivery systems on human IBD and assisting in focusing the future research in this topic, on strategies which could provide maximum remission in IBD patients.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Equilibrium and Release Properties of Hyaluronic acid-Drug complexes.

  Battistini Franco David, Olivera María Eugenia, Manzo Rubén Hilario
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  ABSTRACT: Abstract With the aim to provide more rational basis about the potentiality of hyaluronic acid (or hyaluronan) as drug carrier a set of ionic complexes of its acid form (HA) and its sodium salt (NaHA) with three model drugs (D) (atenolol, propranolol and lidocaine) were prepared. Besides NaHA subjected to hyalurodinase depolimerization (NaHAd) was also used. Transparent dispersions were obtained. They exhibited negative electrokinetic potential and a high degree of counterionic condensation with affinity constants (log Kcc) in the range of 5.8 - 6.1 for propranolol complexes (pKa 9.45) and 4.0 - 4.6 for lidocaine ones (pKa 7.92). Delivery rates of D from the complexes were measured in a Franz-type bicompartimental device. Loaded D were slowly released from the three types of complexes, even when a neutral salt was added to the dispersion placed in the donor compartment, revealing the high affinity between the protonated drugs and the ionisable groups of the polymer. Complex dispersions based on HA or on NaHAd exhibited lower viscosity than those of NaHA but their complexing ability remained unaltered. The results reported on equilibrium and release properties of Hyaluronan-model D complexes contribute to expand the use of HA and NaHA as drug carriers for different routes of administration.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Amphiphilic glycopolymer nanoparticles as vehicles for nasal delivery of peptides and proteins.

  Chao Zheng, Qianqian Guo, Zhongming Wu, Lei Sun, Zhengpu Zhang, Chaoxing Li, Xinge Zhang
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  ABSTRACT: Nasal drug delivery system has been a very promising route for delivery of proteins and peptides for the reason that it can avoid degradation in gastrointestinal tract and metabolism by liver enzymes. However, the bioavailability of proteins and peptides is still low due to the rapid clearance of mucociliary. Here, to prolong the residence time of drugs and improve their absorption, we prepared amphiphilic glycopolymer poly(2-lactobionamidoethyl methacrylate-random-3-acrylamidophenylboronic acid) (p(LAMA-r-AAPBA), and the glycopolymer could assemble into the nanoparticles with narrow size distribution. Insulin, as a model drug, was efficiently encapsulated within the nanoparticles, and loading capacity was up to 12%. In vitro study revealed that the insulin release could be controlled by modifying the composition of glycopolymers. Cell viability showed that p(LAMA-r-AAPBA) nanoparticles had good cytocompatibility. Moreover, the mechanism of nanoparticle internalization into Calu-3 cells was a combination mechanism of clathrin-mediated endocytosis and lipid raft/caveolae-mediated endocytosis. Importantly, there was a significant decrease in the blood glucose levels after the nasal administration of p(LAMA-r-AAPBA) nanoparticles to diabetic rats. Therefore, p(LAMA-r-AAPBA) glycopolymers have a potential application as a nasal delivery systems for proteins and peptides.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: The role of hepatic transport and metabolism in the interactions between pravastatin or repaglinide and two rOatp inhibitors in rats.

  Lassina Badolo, Christoffer Bundgaard, Mats Garmer, Bente Jensen
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  ABSTRACT: A change in the function or expression of hepatic drug transporters may have significant effect on the efficacy or safety of orally administered drugs. Although a number of clinical drug-drug interactions associated with hepatic transport proteins have been reported, in practice it is not always straightforward to discriminate other pathways (e.g. drug metabolism) from being involved in these interactions. The present study was designed to assess the interactions between organic anion transporting polypeptide (Oatp) substrates (pravastatin or repaglinide) and inhibitors (spironolactone or diphenhydramine) in vivo in rats. The mechanisms behind the interactions were then investigated using in vitro tools (isolated hepatocytes and rat liver microsomes). The results showed a significant increase in the systemic exposures of pravastatin (2.5 fold increase in AUC) and repaglinide (1.8 fold increase in AUC) after co-administration of spironolactone to rats. Diphenhydramine increased the AUC of repaglinide by 1.4 fold. The in vivo interactions observed in rats between Oatp substrates and inhibitors may a priori be classified as transport-mediated drug-drug interactions. However, mechanistic studies performed in vitro using both isolated rat hepatocytes and rat liver microsomes showed that the interaction between pravastatin and spironolactone may be solely linked to the inhibition of pravastatin uptake in liver. On the contrary, the inhibition of cytochrome P450 seemed to be the reason for the interactions observed between repaglinide and spironolactone. Although the function and structure of transport proteins may vary between rats and humans, the approach used in the present study can be applied to humans and help to understand the role of drug transport and drug metabolism in a given drug-drug interaction. This is important to predict and mitigate the risk of drug-drug interactions for a candidate drug in pre-clinical development, it is also important for the optimal design of drug-drug interactions studies in the clinic.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Glycerol and urea can be used to increase skin permeability in reduced hydration conditions.

  Sebastian Björklund, Johan Engblom, Krister Thuresson, Emma Sparr
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  ABSTRACT: The natural moisturizing factor (NMF) is a group of hygroscopic molecules that is naturally present in skin and protects from severe drying. Glycerol and urea are two examples of NMF components that are also used in skin care applications. In the present study, we investigate the influence of glycerol and urea on the permeability of a model drug (metronidazole, Mz) across excised pig skin membranes at different hydrating conditions. The degree of skin hydration is regulated by the gradient in water activity across the membrane, which in turn depends on the water activity of the formulation in contact with the skin membrane. Here, we determine the water activity of all formulations employed using an isothermal calorimetric method. Thus, the gradient in water activity is controlled by a novel experimental set-up with well-defined boundary conditions on both sides of the skin membrane. The results demonstrate that glycerol and urea can retain high steady state flux of Mz across skin membranes at dehydrating conditions, which otherwise would decrease the permeability due to dehydration. X-ray diffraction measurements are performed to give insight into the effects of glycerol and urea on SC molecular organization. The novel steady state flux results can be related to the observation that water, glycerol, and urea all affect the structural features of the SC molecular components in a similar manner.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Monitoring of imatinib targeted delivery in human leukocytes.

  Eva Kralj, Simon Zakelj, Jurij Trontelj, Tadej Pajič, Irena Preložnik Zupan, Peter Cernelč, Barbara Ostanek, Janja Marc, Albin Kristl
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  ABSTRACT: The success of imatinib therapy in chronic myeloid leukemia is highly influenced by its active transport into target cells. However, the methodology for analytical evaluation of intracellular drug concentration is rare and usually reliant upon the use of radioactively labeled drugs. More specifically, there is no published method available in the literature for the determination of imatinib concentration in granulocytes. To gain further insight into the intracellular drug uptake a very reliable two-stage sample concentration procedure was devised and coupled with a sensitive ultra-high performance liquid chromatography tandem mass spectrometry. The reliability of this sample preparation and sensitivity of the analysis was confirmed by a successful validation of all necessary method parameters to an impressive lower limit of quantification of 0.5 ng imatinib per 10(6) cells still at the signal to noise ratio of 670. The usefulness of the method is further improved with only 6 mL of blood being necessary for patient analysis. The method has been applied to blood samples of 13 CML patients treated with imatinib and all the measured intracellular drug concentrations were within the validated range. These and further measurements will enable the research of factors which may, besides blood plasma concentration, influence the individual's response to imatinib therapy. Furthermore, individualisation of dosing based on the directly measured targeted drug delivery could be possible.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Ion-pair strategy for enabling Amifostine oral absorption: rat in situ and in vivo experiments.

  N Samiei, V Mangas-Sanjuan, I González-Álvarez, M Foroutan, A Shafaati, A Zarghi, M Bermejo
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  ABSTRACT: This study shows the effect of ion pair formation on intestinal absorption and oral bioavailability of amifostine. Amifostine is a prodrug used as a highly potent and selective radiotherapy and chemotherapy protectant but due to its low lipophilicity and charge at physiological pH range, its trans epithelial transport and its potential for oral drug delivery is very low. Ion pair formation with negatively charged counter ions was evaluated by in situ rat perfusion studies as a possible strategy to enhance intestinal absorption of amifostine. Succinic acid, phthalic acid and benzoic acid were used as counter ions. Rat intestinal perfusion studies confirmed a statistically significant increase in amifostine permeability in the presence of the counter ions in the order of succinic > phthalic > benzoic. Rat pharmacokinetic studies in vivo were performed to calculate oral absolute bioavailability of amifostine alone and with ion pairs in order to confirm the in situ perfusion results and the applicability of the ion pair approach. Intravenous and intraduodenal administrations were done in rats using a permanent jugular vein cannulation technique and a duodenal cannulation method to avoid drug degradation in stomach. In vivo oral bioavailability studies demonstrated a 20-30 fold increase in amifostine bioavailability with succinic acid depending on counter ion ratio and 10 fold increase with phthalic acid as ion pair. In summary ion pair strategy with succinic acid could enable amifostine oral administration on enteric coated formulations.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Medicines Combinations Options and Regulatory Hurdles.

  Ole J Bjerrum, Yvonne Gautam, Esben J Bjerrum, Merete Schmiegelow, Harrie C M Boonen
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  ABSTRACT: The number of fixed dose combinations (FDC) on the market is increasing. However, both regulatory guidelines and the evidence for better effectiveness of fixed dose combinations are not established yet. These aspects were the topic for a recent EUFEPS workshop in Copenhagen. The discussion confirms the presence of hurdles and recommendations to further progress and alleviate the problems. The general conclusion was: Refinement of general systems pharmacology tools for identification of key molecular targets in disease development; Precompetitive methodologies and technologies designed for (co-) development of FDCs; Simulation and modeling algorithms applied on pharmacodynamics (synergy), pharmacokinetics and safety; New formulations of FDCs of solid, liquid or inhalation forms; Prospective clinical studies for measuring the efficacy and effectiveness of use of drug-drug combinations in elderly.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: 1,4-Substituted 4-(1H)-pyridylene-hydrazone-type inhibitors of AChE, BuChE, and amyloid-β aggregation crossing the blood-brain barrier.

  Michaela Prinz, Sülünay Parlar, Gül Ah Bayraktar, Vildan Alptüzün, Ercin Erciyas, Adyary Fallarero, Daniela Karlsson, Pia Vuorela, Malgorzata Burek, Carola Förster, Ezgi Turunc, Guliz Armagan, Ayfer Yalcin, Carola Schiller, Kristina Leuner, Manuel Krug, Christoph A Sotriffer, Ulrike Holzgrabe
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  ABSTRACT: Given the fundamentally multifactorial character of Alzheimer's disease (AD), addressing more than one target for disease modification or therapy is expected to be highly advantageous. Here, following the cholinergic hypothesis, we aimed to inhibit both acetyl- and butyrylcholinesterase (AChE and BuChE) in order to increase the concentration of acetylcholine in the synaptic cleft. In addition, the formation of the amyloid fibrils should be inhibited and already preformed fibrils should be destroyed. Based on a recently identified AChE inhibitor with a 1,4-substituted 4-(1H)-pyridylene-hydrazone skeleton, a substance library has been generated and tested for inhibition of AChE, BuChE, and fibril formation. Blood-brain barrier mobility was ensured by a transwell assay. Whereas the p-nitrosubstituted compound 18C shows an anti-AChE activity in the nanomolar range of concentration (IC50 = 90 nM), the bisnaphthyl substituted compound 20L was found to be the best overall inhibitor of AChE/BuChE and enhances the fibril destruction.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Lecithin based Lamellar liquid crystals as a physiologically acceptable dermal delivery system for Ascorbyl Palmitate.

  Mirjam Gosenca, Marija Bešter Rogač, Mirjana Gašperlin
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  ABSTRACT: Liquid crystalline systems with a lamellar structure have been extensively studied as dermal delivery systems. Ascorbyl palmitate (AP) is one of the most studied and used ascorbic acid derivatives and is employed as an antioxidant to prevent skin aging. The aim of this study was to develop and characterize skin-compliant dermal delivery systems with a liquid crystalline structure for AP. First, a pseudoternary phase diagram was constructed using Tween 80/lecithin/isopropyl myristate/water at a Tween 80/lecithin mass ratio of 1/1, and the region of lamellar liquid crystals was identified. Second, selected unloaded and AP-loaded lamellar liquid crystal systems were physicochemically characterized with polarizing optical microscopy, small-angle X-ray scattering, differential scanning calorimetry, and rheology techniques. The interlayer spacing and rheological parameters differ regarding quantitative composition, whereas the microstructure of the lamellar phase was affected by the AP incorporation, resulting either in additional micellar structures (at 25 and 32°C) or being completely destroyed at higher temperature (37°C). After this, the study was oriented towards in vitro cytotoxicity evaluation of lamellar liquid crystal systems on a keratinocyte cell line. The results suggest that the lamellar liquid crystals that were developed could be used as a physiologically acceptable dermal delivery system.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 05/2013;
• Article: Application of COSMO-RS as an Excipient Ranking Tool in Early Formulation Development.

  Agnieszka Pozarska, Claudia da Costa Mathews, Mei Wong, Klimentina Pencheva
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  ABSTRACT: The low amounts of drug available in early discovery often results in limited information on the physico-chemical (solubility etc.) properties of a compound being obtained. As a result, predictive tools and miniaturised screens have been investigated to aid formulation development in early discovery. This study looks at the potential application of the quantum chemistry program, Conductor Screening Model for Real Solvents (COSMO-RS) to help with the selection of excipients for formulation development in early discovery. The excipient solubility predictions obtained from COSMO-RS were compared to experimentally obtained solubilities. The results showed that in general, COSMO-RS was able to help formulators with the selection of the most appropriate excipients to solubilise the model compound.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;
• Article: Intracellular trafficking of solid lipid nanoparticles and their distribution between cells through tunneling nanotubes.

  Julijana Kristl, Karmen Teskač Plajnšek, Mateja Erdani Kreft, Biljana Janković, Petra Kocbek
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  ABSTRACT: The intracellular fate of nanosized drug delivery systems is still not well understood. Various internalization pathways have been discovered, but knowledge of their intracellular trafficking is still incomplete. The aim of this study was to examine the internalization, pathways, and positioning taken by solid lipid nanoparticles (SLNs) in cells. SLNs were fluorescence labelled with a newly synthesized fluorescent probe, 14-DACA. The probe was strongly incorporated into the nanoparticle core under the influence of its long lipophilic chain, enabling superior visualization of SLNs under complex and dynamic intracellular conditions. The intracellular distribution of SLNs was studied qualitatively using a co-localization technique and quantitatively using fluorescence intensity profiles. SLNs were seen inside the cells as distinct bright blue dots that underwent dynamic movement and were finally positioned in the proximity of the nucleus. A few SLNs were shown to be present in mitochondria and between actin filaments, but none in the cell nucleus or lysosomes. SLNs are here reported to be present in tunneling nanotubes (TNTs), which could be a new route of SLN transfer between cells. More TNTs were observed in cells treated with SLNs. The presence of TNTs was additionally confirmed by atomic force microscopy analysis, which indicated that treated cells were more rough than control cells. Detailed investigation of the subcellular localization of SLNs and the evidence for their transfer and distribution via TNTs to the cells, which are not in direct contact with the source of SLNs, are important for understanding the mechanism of targeted drug delivery. Understanding the possible intercellular distribution of SLNs via TNTs can significantly influence approaches to treating organelle-specific diseases.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;
• Article: Preparation of isoliquiritigenin-loaded nanostructured lipid carrier and the in vivo evaluation in tumor-bearing mice.

  Xiao Yun Zhang, Hua Qiao, Jing Man Ni, Yan Bin Shi, Yin Qiang
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  ABSTRACT: Isoliquiritigenin-loaded nanostructured lipid carrier (ISL-NLC) was constructed and characterized. In vivo antitumor efficacy and immuno-modulation effects of ISL-NLC were evaluated in sarcoma 180 (S180)-bearing and murine hepatoma 22 (H22)-bearing mice model through intraperitoneal (i.p.) administration. The ISL-NLC biodistribution was also investigated in H22-bearing mice. Results demonstrated that the ISL-NLC had a spherical shape with a mean size of (160.73 ± 6.08) nm and encapsulation efficiency of (96.7 4 ± 1.81) %. ISL released from the nanoparticles was in a sustained manner with an initial burst release. ISL-NLC significantly inhibit tumor growth at 10, 20 and 40 mg/kg levels, and inhibition rates were 75.70, 82.27 and 83.90% in the S180-bearing mice and 71.49, 81.11 and 85.62% in the H22-bearing mice, respectively. The biodistribution study showed that ISL concentration of ISL-NLC in tumor is higher 2.5-fold than ISL suspension. The elimination half-life (t1/2), area under the curve (AUC) and the mean residence times (MRT) of the ISL-NLC was much longer than that of the ISL suspension. As a whole, anticancer effect of ISL encapsulated in NLC was superior to ISL in suspension on H22-bearing and S180-bearing mice at the same dose and was a dose-dependent way, and ISL-NLC improved immunity of ISL. It can be inferred that nanostructured lipid carriers are a promising carrier for cancer therapy using ISL.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;
• Article: Viscosity of high concentration protein formulations of monoclonal antibodies of the IgG1 and IgG4 subclass - prediction of viscosity through protein-protein interaction measurements.

  Martin S Neergaard, Devendra S Kalonia, Henrik Parshad, Anders D Nielsen, Eva H Møller, Marco van de Weert
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  ABSTRACT: The purpose of this work was to explore the relation between protein-protein interactions (PPI) and solution viscosity at high protein concentration using three monoclonal antibodies (mAbs), two of the IgG4 subclass and one of the IgG1 subclass. A range of methods was used to quantify the PPI either at low concentration (interaction parameter (kD) obtained from dynamic light scattering, DLS) or at high concentration (solution storage modulus (G') from ultrasonic shear rheology). We also developed a novel method for the determination of PPI using the apparent radius of the protein at either low or high protein concentration determined using DLS. The PPI measurements were correlated with solution viscosity (measured by DLS using polystyrene nanospheres and ultrasonic shear rheology) as a function of pH (4 to 9) and ionic strength (10, 50 and 150 mM). Our measurements showed that the highest solution viscosity was observed under conditions with the most negative kD, the highest apparent radius and the lowest net charge. An increase in ionic strength resulted in a change in the nature of the PPI at low pH from repulsive to attractive. In the neutral to alkaline pH region the mAbs behaved differently with respect to increase in ionic strength. Two mAbs (A and B) showed little or no effect of increasing ionic strength, whereas mAb-C showed a remarkable decrease in attractive PPI and viscosity. Previous studies have mainly investigated mAbs of the IgG1 and IgG2 subclass. We show here, for the first time, that mAbs of the IgG4 subclass behave similar as the other subclasses. By comparison of the three tested mAbs with mAbs investigated in other studies a clear linear trend emerges between the pH of strongest attractive PPI and highest solution viscosity. The determination of PPI using either kD or apparent radius is thus a useful prediction tool in the determination of solution conditions that favors low solution viscosity at high protein concentration of therapeutically used mAb molecules. The novel methodology using apparent radius is a simple and rapid alternative to determine relative PPI directly under formulation conditions. The method can potentially serve as a high-throughput screening tool in formulation development.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;
• Article: Inhibition of antigen-induced airway inflammation and hyperresponsiveness in guinea pigs by a selective antagonist of "chemoattractant receptor homologous molecule expressed on Th2 cells" (CRTH2).

  M Tasaki, M Kobayashi, Y Tenda, S Tsujimoto, S Nakazato, M Numazaki, Y Hirano, H Matsuda, T Terasaka, Y Miyao, Y Shimizu, Y Hirayama
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  ABSTRACT: Chemoattractant receptor homologous molecule expressed on T helper type 2 cells (CRTH2) is a PGD2 receptor found on eosinophils, basophils, and Th2 type T cells which exhibits chemotaxis and functions in activation cascades. However, while a number of CRTH2 antagonists, including ramatroban, are known to exert activity in certain animal models, activity in a guinea pig model of EA-induced airway hyperresponsiveness has not been demonstrated. The newly developed CRTH2 antagonist ASP5642 has shown antagonistic activity against human and guinea pig CRTH2 in previous studies and has also been found effective in treating guinea pig models of airway inflammation and airway hyperresponsiveness. While previous studies have used animals such as rats and mice to evaluate CRTH2 antagonist effects, ours is the first attempt to evaluate CRTH2 function in a guinea pig asthma model, which may prove useful in evaluating the compound's effects in humans, given the comparable airway function between the two species Taken together, these data from the present study strongly suggest the utility of ASP5642 in investigating the role of CRTH2 in inflammatory responses and as a drug treatment for human asthma.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;
• Article: Pullulan-based nanoparticles as carriers for transmucosal protein delivery.

  Marita Dionísio, Clara Cordeiro, Carmen Remuñán-López, Begoña Seijo, Ana M Rosa da Costa, Ana Grenha
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  ABSTRACT: Polymeric nanoparticles have revealed very effective in transmucosal delivery of proteins. Polysaccharides are among the most used materials for the production of these carriers, owing to their structural flexibility and propensity to evidence biocompatibility and biodegradability. In parallel, there is a preference for the use of mild methods for their production, in order to prevent protein degradation, ensure lower costs and easier procedures that enable scaling up. In this work we propose the production of pullulan-based nanoparticles by a mild method of polyelectrolyte complexation. As pullulan is a neutral polysaccharide, sulfated and aminated derivatives of the polymer were synthesized to provide pullulan with a charge. These derivatives were then complexed with chitosan and carrageenan, respectively, to produce the nanocarriers. Positively charged nanoparticles of 180-270 nm were obtained, evidencing ability to associate bovine serum albumin, which was selected as model protein. In PBS pH 7.4, pullulan-based nanoparticles were found to have a burst release of 30% of the protein, which maintained up to 24h. Nanoparticle size and zeta potential were preserved upon freeze-drying in the presence of appropriate cryoprotectants. A factorial design was approached to assess the cytotoxicity of raw materials and nanoparticles by the metabolic test MTT. Nanoparticles demonstrated to not cause overt toxicity in a respiratory cell model (Calu-3). Pullulan has, thus, demonstrated to hold potential for the production of nanoparticles with an application in protein delivery.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;
• Article: HQS-3, a newly synthesized flavonoid, possesses potent anti-tumor effect in vivo and in vitro.

  Yuxin Zhou, Na Lu, Haiwei Zhang, Libin Wei, Lei Tao, Qinsheng Dai, Li Zhao, Biqi Lin, Qilong Ding, Qinglong Guo
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  ABSTRACT: HQS-3 is a newly baicalein derivative with a benzene substitution. We investigated the anticancer effect of HQS-3 in vivo and in vitro. HQS-3 significantly decreased tumor growth in mice inoculated with Heps and HepG2 cells; and had little influence on the state and weight of animals. After treatment with 20 mg/kg HQS-3, the inhibitory rate of tumor weight in mice inoculated with Heps and HepG2 cells were 63.62% and 68.03%, respectively. Meanwhile, HQS-3 inhibited the viability of various kinds of tumor cells with IC50 values in the range of 22.98 to 54.32 μM after 48 h treatment measured by MTT-assay. HQS-3 remarkably inhibited viability of hepatoma cells in a concentration- and time-dependent manner and induced apoptosis in HepG2 cells by DAPI staining and Annexin V/PI double staining. The apoptosis-induction effect of HQS-3 was attributed to its ability to modulate the actvity of caspase-9, caspase-3 and PARP. Moreover, the expression of bax protein was increased while the bcl-2 protein was decreased, leading to an increase in Bax/Bcl-2 ratio. The accumulation of ROS induced by HQS-3 in HepG2 cells was also observed. The further results suggested that HQS-3 induced mitochondrial-mediated apoptosis by increasing ROS level and inhibiting the expression of anti-oxidative protein SOD2. HQS-3 exerted anti-tumor activity both in vitro and in vivo via inducing tumor cells apoptosis, and these results suggested that it deserves further investigation as a novel chemotherapy for human tumors.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;
• Article: Tandutinib (MLN518/CT53518) targeted to stem-like cells by inhibiting the function of ATP-binding cassette subfamily G member 2.

  Xiao-Qin Zhao, Chun-Ling Dai, Shinobu Ohnuma, Yong-Ju Liang, Wen Deng, Jun-Jiang Chen, Mu-Sheng Zeng, Suresh V Ambudkar, Zhe-Sheng Chen, Li-Wu Fu
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  ABSTRACT: Tandutinib is a novel inhibitor of tyrosine kinases FLT3, PDGFR and KIT. Our study was to explore the capability of tandutinib to reverse ABC transporter-mediated multidrug resistance. Tandutinib reversed ABCG2-mediated drug resistance in ABCG2-482-R2, ABCG2-482-G2, ABCG2-482-T7 and S1-M1-80 cells and increased the accumulation of doxorubicin, rhodamine 123 and [H(3)] mitoxantrone in ABCG2-overexpressing cells. Importantly, tandutinib selectively sensitized side population cells to mitoxantrone. Taken together, our results advocate the potency of tandutinib as an ABCG2 modulator and stem-like cells targeted agent to increase efficiency of anticancer drugs.
  European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences 04/2013;