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Journal of Pharmaceutical Sciences
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  Hybrid Journal Hybrid journal (It can contain Open Access articles)
ISSN (Print) 0022-3549 - ISSN (Online) 1520-6017
Published by Elsevier Homepage  [3184 journals]
  • Co-processed Particles: An Approach to Transform Poor Tableting Properties
    • Abstract: Publication date: Available online 21 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Rahul Roopwani, Ira S. Buckner The role of co-processing in improving tablet mechanical properties was investigated in this work. Gabapentin was used as the model compound owing to its poor tableting properties such as low tensile strength, strain rate sensitivity, high ejection force and tablet capping. Gabapentin was blended with HPMC (Methocel®) in a high shear mixer to obtain an interactive mixture consisting of finer HPMC particles adsorbed onto the surface of larger gabapentin particles. Interactive mixture containing
  • An Assessment of the Ability of Submicron- and Micron-Size Silicone Oil
           Droplets in Dropped Prefillable Syringes to Invoke Early- and Late-Stage
           Immune Responses
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Elena Krayukhina, Masami Yokoyama, Kayoko Kakuhou Hayashihara, Takahiro Maruno, Masanori Noda, Hiroki Watanabe, Takayuki Uchihashi, Susumu Uchiyama A number of biopharmaceuticals are available as lyophilized formulations along with a prefilled syringe (PFS) containing water for injection (WFI). Submicron- and micron-size droplets of lubricating silicone oil (SO) applied to the inner surface of the PFS barrel might migrate into the WFI, to which protein pharmaceuticals can adsorb, potentially inducing an immune response. In the present study, we subjected siliconized cyclo-olefin polymer PFSs filled with WFI to dropping stress to simulate actual shipping conditions as well as evaluated the risk associated with the released SO droplets. The results confirmed the undesirable effects of SO on therapeutic proteins, including adsorption to SO droplets and increased secretion of several innate cytokines from human peripheral blood mononuclear cells of a small donor panel. Assessment of immunogenicity in vivo using BALB/c mice revealed a slight increase in the plasma concentrations of antidrug antibodies over 21 days in response to SO-containing antibody samples compared to the absence of SO. These results indicate that SO droplets form complexes with pharmaceutical proteins that can potentially invoke early- and late-stage immune responses. Therefore, the use of SO-free cyclo-olefin polymer PFSs as primary containers for WFI could contribute to the enhanced safety of reconstituted biopharmaceuticals.
  • In Vitro-In Vivo Extrapolation and Hepatic Clearance-Dependent
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Christine M. Bowman, Leslie Z. Benet Accurately predicting the hepatic clearance of compounds using in vitro to in vivo extrapolation (IVIVE) is crucial within the pharmaceutical industry. However, several groups have recently highlighted the serious error in the process. Although empirical or regression-based scaling factors may be used to mitigate the common underprediction, they provide unsatisfying solutions because the reasoning behind the underlying error has yet to be determined. One previously noted trend was intrinsic clearance-dependent underprediction, highlighting the limitations of current in vitro systems. When applying these generated in vitro intrinsic clearance values during drug development and making first-in-human dose predictions for new chemical entities though, hepatic clearance is the parameter that must be estimated using a model of hepatic disposition, such as the well-stirred model. Here, we examine error across hepatic clearance ranges and find a similar hepatic clearance-dependent trend, with high clearance compounds not predicted to be so, demonstrating another gap in the field.
  • Validation of Human MDR1-MDCK and BCRP-MDCK Cell Lines to Improve the
           Prediction of Brain Penetration
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Bo Feng, Mark West, Nandini C. Patel, Travis Wager, Xinjun Hou, Jillian Johnson, Larry Tremaine, Jennifer Liras It is of great challenge to predict human brain penetration for substrates of multidrug resistance protein 1 (MDR1) and breast cancer resistance protein (BCRP), 2 major efflux transporters at blood-brain barrier. Thus, a physiologically based pharmacokinetic (PBPK) model with the incorporation of in vitro MDR1 and BCRP transporter function data and transporter protein expression levels has been developed. As such, it is crucial to generate MDR1 and BCRP substrate data with a high fidelity. In this study, 2 widely used human MDR1 cell lines from Borst and National Institutes of Health laboratories were evaluated using rodent brain penetration data, and the study suggested that the MDR1 expressed in Madin-Darby canine kidney (MDCK) cell line from National Institutes of Health laboratory predicted brain penetration better, particularly for compounds with a high passive permeability. In addition, human BCRP-MDCK cell line with 1 μM PSC833, a specific MDR1 inhibitor, demonstrated the ability to identify BCRP substrates without the confounding of endogenous canine Mdr1. Comparison of human BCRP and mouse Bcrp transporter functions revealed that the functional differences of BCRP between the 2 species is minimal. The incorporation of both the validated MDR1 and BCRP assays into our brain PBPK model has significantly improved the prediction for the brain penetration of MDR1 and BCRP substrates across species.
  • A “Dual” Cell-Level Systems PK-PD Model to Characterize the
           Bystander Effect of ADC
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Aman P. Singh, Dhaval K. Shah Here, we have developed a cell-level systems PK-PD model to characterize the bystander effect of antibody-drug conjugates (ADCs). Cytotoxicity data generated following incubation of Trastuzumab-vc-MMAE in cocultures of high HER2-expressing N87 and low HER2-expressing GFP-MCF7 cells were used to build the model. Single-cell PK model for ADC was used to characterize the PK of trastuzumab-vc-MMAE and released MMAE in N87 and GFP-MCF7 cells. The 2 cell-level PK models were mechanistically integrated to mimic the coculture condition. MMAE-induced intracellular occupancy of tubulin was used to drive the efficacy of ADC, and improvement in the tubulin occupancy of GFP-MCF7 cells in the presence of N87 cells was used to drive the bystander effect of trastuzumab-vc-MMAE. The “dual” cell-level PK-PD model was able to capture the observed data reasonably well. It was found that similar and high occupancy of tubulin by MMAE was required to achieve the cytotoxic effect in each cell line. In addition, estimated model parameters suggested that ∼60% improvement in the tubulin occupancy was required to attain half of the maximum bystander killing effect by the ADC. The presented model provides foundation for in vivo systems PK-PD model to characterize and predict the bystander effect of ADCs.
  • Solid-State Characterization of Spironolactone 1/3 Hydrate
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Thúlio Wliandon Lemos Barbosa, Antônio Carlos Doriguetto, Magali Benjamim de Araújo, Rudy Bonfilio Spironolactone (SPR) is a poorly water-soluble drug widely used for the treatment of various diseases. The objective of this study was to carry out the preparation and solid-state characterization of SPR 1/3 hydrate. The solid form was generated by an unreported recrystallization process in acetone and characterized for the first time by a combination of X-ray powder diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), Fourier-transform infrared spectroscopy (FTIR), equilibrium solubility, and an accelerated stability study. XRD, DSC, and TGA studies revealed that SPR 1/3 hydrated converts completely to form II after heating to 180°C. Solubility studies at 37°C showed that SPR 1/3 hydrate was statistically less soluble than SPR form II in all tested media and that SPR form II partially converts to SPR 1/3 hydrate in aqueous media. Accelerated stability studies demonstrated that both forms were physically and chemically stable up to 6 months (40°C/75% RH). We concluded that contamination of SPR 1/3 hydrate in SPR raw materials is undesirable. Taking this into account we recommend its polymorphic monitoring either in active pharmaceutical ingredients or commercial tablets by solid-state identification/quantification methods (XRD, DSC, TGA, and FTIR). Of these, XRD proved to be the most conclusive and accurate.
  • Freeze-Dried Matrices Based on Polyanion Polymers for Chlorhexidine Local
           Release in the Buccal and Vaginal Cavities
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Barbara Giordani, Angela Abruzzo, Umberto Maria Musazzi, Francesco Cilurzo, Fiore Pasquale Nicoletta, Francesco Dalena, Carola Parolin, Beatrice Vitali, Teresa Cerchiara, Barbara Luppi, Federica Bigucci Chlorhexidine (CLX) is a wide spectrum cationic antimicrobial used for prevention and treatment of infections of buccal and vaginal cavities. To increase the residence time of CLX-based formulations at the application site and consequently reduce the daily dose frequency, new formulations composed of mucoadhesive polymers should be designed. The objective of this work was the development of matrices based on polyanionic polymers, such as sodium alginate, carboxymethylcellulose, xanthan gum and sodium hyaluronate, aimed to prolong the local release of CLX into the buccal or vaginal cavity. Matrices were prepared by freeze-drying and comply with 2 different preparative methods and characterized in terms of resistance to compression, water uptake ability, mucoadhesion, in vitro drug release behavior and antimicrobial activity toward representative pathogens of buccal and vaginal cavities. Results showed that the selection of suitable polymers associated to the adequate preparative method allowed to modulate matrix ability to hydrate, adhere to the mucosa and release the drug as well as to exert antimicrobial activity. In particular, matrix based on sodium hyaluronate was found to be the best performing formulation and could represent a versatile system for local release of CLX with potential application in both buccal and vaginal cavities.
  • Lactoferrin-Loaded Alginate Microparticles to Target Clostridioides
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Shwana Braim, Klaudyna Śpiewak, Malgorzata Brindell, Daniela Heeg, Cameron Alexander, Tanya Monaghan Some forms of bovine lactoferrin (bLf) are effective in delaying Clostridioides difficile growth and preventing toxin production. However, therapeutic use of bLf may be limited by protein stability issues. The objective of this study was to prepare and evaluate colon-targeted, pH-triggered alginate microparticles loaded with bioactive bLf and to evaluate their anti-C difficile defense properties in vitro. Different forms of metal-bound bLf were encapsulated in alginate microparticles using an emulsification or internal gelation method. The microparticles were coated with chitosan to control protein release. In vitro drug release studies were conducted in pH-simulated gastrointestinal conditions to investigate the release kinetics of encapsulated protein. No significant release of metal-bound bLf was observed at acidic pH; however, on reaching simulated colonic pH, most of the encapsulated lactoferrin was released. The application of bLf (5 mg/mL) delivered from alginate microparticles to human intestinal epithelial cells significantly reduced the cytotoxic effects of toxins A and B as well as bacterial supernatant on Caco-2 and Vero cells, respectively. These results are the first to suggest that alginate-bLf microparticles show protective effects against C difficile toxin-mediated epithelial damage and impairment of barrier function in human intestinal epithelial cells. The future potential of lactoferrin-loaded alginate microparticles against C difficile deserves further study.
  • Tuning Drug Release via Twin Screw Extrusion in Polyester Films
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Yubin Zhou, Devante Horne, Terry W.J. Steele Laboratory investigations of polyester biomaterials overly rely on additives and solvent-casting manufacturing techniques that do not allow translation into industrial processing owing to the volatile organic compounds present. These impediments of bench to bedside translation are addressed with a “green chemistry” twin screw extrusion approach, where the extrusion parameters control the release profile. Advantages include elimination of volatile organic compounds, removal of plastic plasticizers/additives, and a low temperature production that is applicable to both academia and industry. Poly (d, l-lactide-co-glycolide) (PLGA), a labile polyester drug depot is loaded with fluorescein diacetate (FDAc), a pH/heat sensitive drug that degrades to fluorescein. Processing of FDAc suggests no degradation at elevated temperatures or high shears, indicating hydrophobic small molecules can be homogeneously dispersed with no detrimental effects. In vitro release of FDAc in physiological environments varies release profile through ratios of ester/acid-terminated PLGA without secondary additives. Postprocessing stability investigations of PLGA resin with size exclusion chromatography and differential scanning calorimetry validate no significant polymer degradation. Through judicious choice of parameters, release can be tuned from 15-250 microgram per sq. cm per day. Twin screw extrusion production of PLGA drug depots reveals a promising method of tailor-made drug delivering thin films for various pharmaceutical applications.
  • Mannosylated Nanoparticles for Oral Immunotherapy in a Murine Model of
           Peanut Allergy
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Ana Brotons-Canto, Carlos Gamazo, Nekane Martín-Arbella, Muthanna Abdulkarim, Mark Gumbleton, Gemma Quincoces, Ivan Peñuelas, Juan M. Irache Peanut allergy is one of the most prevalent and severe of food allergies with no available cure. The aim of this work was to evaluate the potential of an oral immunotherapy based on the use of a roasted peanut extract encapsulated in nanoparticles with immunoadjuvant properties. For this, a polymer conjugate formed by the covalent binding of mannosamine to the copolymer of methyl vinyl ether and maleic anhydride was first synthetized and characterized. Then, the conjugate was used to prepare nanoparticles with an important capability to diffuse through the mucus layer and reach, in a large extent, the intestinal epithelium, including Peyer’s patches. Their immunotherapeutic potential was evaluated in a model of presensitized CD1 mice to peanut. After completing therapy, mice underwent an intraperitoneal challenge with peanut extract. Nanoparticle-treatment was associated with both less serious anaphylaxis symptoms and higher survival rates than control, confirming the protective effect of this formulation against the challenge.
  • Development of Pyro-Drive Jet Injector With Controllable Jet Pressure
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Hiroshi Miyazaki, Shingo Atobe, Takamasa Suzuki, Hiromitsu Iga, Kazuhiro Terai Various jet injectors have been developed and used for the effective and efficient administration of drugs. Jet injections overcome the limitations of other drug delivery methods, such as ablation, iontophoresis, electroporation, sonophoresis, and microneedles, because jet injection is not limited by the diffusion rates of different drugs. However, controlling the jet pressure during drug delivery is difficult with most conventional jet injectors. Efficacy evaluation of such devices on laboratory animals is strongly required before initiating human clinical trials, but minimal research has been performed for the device developments. Therefore, we developed jet injector devices based on pyrotechnics using 2 types of explosives with different burning rates; we call these pyro-drive jet injectors. The liquid jet pressure profile suggests that the penetration depth and injection volume for soft materials and skin tissue are controllable. Here, we propose the pyro-drive jet injectors as another candidate well-controlled jet injector for laboratory animals in drug discovery testing as well as human clinical use.
  • Contribution of Intravenous Administration Components to Subvisible and
           Submicron Particles Present in Administered Drug Product
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Mark Pollo, Anjali Mehta, Kate Torres, Daniel Thorne, Daniel Zimmermann, Parag Kolhe Particulate matter present in drug products intended for parenteral administration to patients is typically monitored and controlled in the finished drug product to minimize potential risks to patients. In contrast to particulates found in drug products, the current study evaluated particulates representative of materials and operations typically used in the dose preparation and administration of drug products. A comprehensive assessment of intrinsic and extrinsic sources of subvisible and submicron particulates arising from materials associated with subcutaneous and intravenous dose preparation and administration was conducted. In particular, particles arising from disposable syringes, commercial sterile diluents, and intravenous supplies were quantitated using established methods for subvisible (light obscuration, flow imaging) and submicron particles (resistive pulse sensing). Each of these sources contributed varying amounts of particulates; therefore, owing to sources from materials required for administration, it is inadequate to assume that the total particulate load delivered to patients arises solely from the drug product. Careful consideration of the administration method and supplies used can improve the predictability of particulate levels present in dose preparations or administration volumes.
  • Variance Between Different Light Obscuration and Flow Imaging Microscopy
           Instruments and the Impact of Instrument Calibration
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Anja Matter, Atanas Koulov, Satish Singh, Hanns-Christian Mahler, Helena Reinisch, Carolin Langer, Benjamin Zucol, Roman Mathaes Subvisible particles (SVPs) are an obligatory critical quality attribute of the product, and yet, they are found in all biopharmaceutical products intended for infusion or injection. Light obscuration (LO) is the primary pharmacopeial method used to quantify SVPs. However, the method may not be equally sensitive toward all particles that can possibly occur. Calibration of LO instruments is usually performed using polystyrene beads suspended in water. In this study, the dependence of the sizing accuracy of LO analysis was evaluated by using a calibration suspension of lower refractive index beads made of silica suspended in sucrose solution. It was demonstrated that the sizing accuracy was strongly dependent on the reference material’s properties used for calibration. It was also demonstrated that flow imaging microscopy suffered from the same artifact, albeit to a smaller extent. We further tested different LO sensors and instruments. Interestingly, our results show that the sizing accuracy varied from instrument to instrument, strongly depending on the properties of the sensor.To summarize, sizing and counting accuracies were dependent on the material used for calibration and its optical properties as well as the calibration curve, the sensor, and the instrument supplier. Closer match of optical properties between calibration system and test system seems to improve the sensitivity of the measurement. The results of this study raise the following major practical implications: (1) LO and flow imaging microscopy are not truly orthogonal analytical methods, (2) while matching optimal properties of material used for calibration and test items increased sensitivity, this is of poor practical applicability given that analytes contain multiple particles, and (3) setting product-specific limits for SVPs require special considerations with regard to the data sets used.
  • Inhibiting Surface Crystallization and Improving Dissolution of Amorphous
           Loratadine by Dextran Sulfate Nanocoating
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Aiguo Zeng, Xin Yao, Yue Gui, Yuhui Li, Karen J. Jones, Lian Yu Amorphous formulations provide a solution to poor solubility and slow dissolution of many drugs, but fast surface crystallization can negate their advantages. As in the case of many amorphous drugs, loratadine (LTD) shows much faster crystal growth on the free surface than in the bulk, and its surface crystallization can be inhibited by a polymer nanocoating. LTD is a weak base with a pKa of 5.25. Dextran sulfate (DTS), a pharmaceutically acceptable polymer, is deposited on amorphous LTD from coating solution at pH 3.5 at which LTD is positively charged. Zeta potential measurements support the mechanism of nanocoating by electrostatic deposition. DTS nanocoating is as good as gold coating for inhibiting surface crystallization of amorphous LTD and significantly increases its rate of dissolution. The enhanced dissolution is likely a result of improved wetting of amorphous particles by an aqueous medium. These results indicate that fast surface crystallization of amorphous LTD is enabled by high mobility of surface molecules, and an ultrathin nanocoating can immobilize surface molecules and inhibit surface crystallization. This nanocoating technique can be used to stabilize amorphous drugs prone to surface crystallization and improve their dissolution, and DTS is an effective nanocoating material for basic drugs such as LTD.
  • A Novel Drug-Drug Cocrystal of Levofloxacin and Metacetamol: Reduced
           Hygroscopicity and Improved Photostability of Levofloxacin
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Taeko Shinozaki, Makoto Ono, Kenjirou Higashi, Kunikazu Moribe Levofloxacin (LVFX), a broad-spectrum antibacterial agent from the fluoroquinolone family, is universally prescribed with antipyretics, including paracetamol (APAP) analogs. In this study, a new drug-drug cocrystal of LVFX and an APAP analog was developed using a grinding and heating approach. Among 9 APAP analogs, only metacetamol (AMAP) was able to form a cocrystal with LVFX, with a stoichiometric ratio of 1:1. This cocrystal was obtained from a eutectic melt of anhydrous LVFX and AMAP after complete desorption of water from LVFX hemihydrate. The crystal structure of the cocrystal was determined by single-crystal X-ray structural analysis. Unlike LVFX hydrates, the LVFX-AMAP cocrystal did not form a channel structure where water molecules reside in LVFX hydrates. Thus, the LVFX-AMAP cocrystal did not undergo hydration under high relative humidity conditions during vapor sorption-desorption analysis and physical stability tests. LVFX photostability was improved by cocrystallization when compared with that of the hemihydrate because of hydrogen bond formation between the hydroxyl group of AMAP and the N-methylpiperazine group of LVFX, which is possibly involved in LVFX photodegradation. The LVFX-AMAP cocrystal, which is superior to LVFX hydrates in both pharmacological and physicochemical properties, is expected to be a useful solid form.
  • Solid and Solution State Thermodynamics of Polymorphs of Butamben (Butyl
           4-Aminobenzoate) in Pure Organic Solvents
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Michael Svärd, Lai Zeng, Masood Valavi, Gamid Rama Krishna, Åke C. Rasmuson The solubility of butamben has been measured gravimetrically in pure methanol, 1-propanol, 2-propanol, 1-butanol, and toluene over the temperature range 268-298 K. Polymorph transition and melting temperatures, associated enthalpy changes, and the heat capacity of the solid forms and the supercooled melt have been measured by differential scanning calorimetry. Based on extrapolated calorimetric data, the Gibbs energy, enthalpy and entropy of fusion, and the activity of solid butamben (the ideal solubility) have been calculated from below ambient temperature up to the melting point. Activity coefficients of butamben at equilibrium in the different solvents have been estimated from solubility data and the activity of the solid, revealing that all investigated systems exhibit positive deviation from Raoult's law. Solubility data are well correlated by a semiempirical regression model. On a mass basis, the solubility is clearly higher in methanol than in the other solvents, but mole fraction solubilities are very similar across all 5 solvents. The 2 known polymorphs are enantiotropically related, and the transition point is located at 283 K. Polymorph interconversions occur within 0.3 K of the transition point even in the solid state, and the 2 forms exhibit strong similarities in investigated properties.
  • Mechanistic Evaluation of the Opposite Effects on Initial Burst Induced by
           Two Similar Hydrophilic Additives From Octreotide Acetate–Loaded PLGA
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Jiwei Liu, Hao Ren, Yan Xu, Yonglu Wang, Kuntang Liu, Yaping Zhou, Yuqian Wang, Weilan Li, Jinlan Tang, He Huang, Xueming Li The purpose of the present study was to make a detailed comparison of 2 similar additives about their opposite effects on the initial burst of octreotide acetate from poly(lactic-co-glycolic acid) microspheres. We focused on identifying the key factor that influenced the initial burst of microspheres induced by small hydrophilic additives. The apparent reason resulting in such differences was different pore closing rates on the surface of these 2 batches. However, the potential mechanism was still unknown. To compare with the single-additive system, these 2 additives were coencapsulated together into the same formulation. Of surprise, the inhibition effect of glucose on burst release somehow disappeared and even turned out to be opposite. This phenomenon greatly reminds us that there must be some interactions between glucose and polymer, which are likely to be disturbed by coencapsulated CaCl2. However, small amount of additive can hardly make any detected difference. Therefore, additive-loaded microspheres without drug were prepared to further investigate the potential factors. Under this condition, differences were found. The key factor for glucose-induced accelerated pore closure and reduction in initial burst was the formation of hydrogen bonds between the glucose molecule and the polymer matrix.
  • Evaluation of a 3D Human Artificial Lymph Node as Test Model for the
           Assessment of Immunogenicity of Protein Aggregates
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Teresa Kraus, Annika Lubitz, Ulrike Schließer, Christoph Giese, Jana Reuschel, René Brecht, Julia Engert, Gerhard Winter The immunogenicity of protein aggregates has been investigated in numerous studies. Nevertheless, it is still unknown which kind of protein aggregates enhance immunogenicity the most. The ability of the currently used in vitro and in vivo systems regarding their predictability of immunogenicity in humans is often questionable, and results are partially contradictive. In this study, we used a 2D in vitro assay and a complex 3D human artificial lymph node model to predict the immunogenicity of protein aggregates of bevacizumab and adalimumab. The monoclonal antibodies were exposed to different stress conditions such as light, heat, and mechanical stress to trigger the formation of protein aggregates and particles, and samples were analyzed thoroughly. Cells and culture supernatants were harvested and analyzed for dendritic cell marker and cytokines. Our study in the artificial lymph node model revealed that bevacizumab after exposure to heat triggered a TH1- and proinflammatory immune response, whereas no trend of immune responses was seen for adalimumab after exposure to different stress conditions. The human artificial lymph node model represents a new test model for testing the immunogenicity of protein aggregates combining the relevance of a 3D human system with the rather easy handling of an in vitro setup.
  • Cocrystal and Salt Forms of an Imidazopyridazine Antimalarial Drug Lead
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Terence J. Noonan, Kelly Chibale, Peter M. Cheuka, Susan A. Bourne, Mino R. Caira Cocrystallization and salt formation were used to produce new multicomponent forms of a novel antimalarial imidazopyridazine drug lead (MMV652103) that displayed improved physicochemical properties. The drug lead had earlier shown good in vitro potency against multidrug resistant (K1) and sensitive (NF54) strains of the human malaria parasite Plasmodium falciparum, and high in vivo efficacy in both Plasmodium berghei and Plasmodium falciparum mouse models. A major drawback of MMV652103 is its limited aqueous solubility. Various new supramolecular products, including several multicomponent solid forms, are reported here, namely 3 cocrystal forms with the dicarboxylic acid coformers adipic acid, glutaric acid, and fumaric acid, and a salt form with malonic acid. These were characterized by thermal methods and their structures elucidated by single-crystal X-ray diffraction. A customized solubility experiment was performed in fasted-state simulated intestinal fluid for comparison of the solubility behavior of each new form of the drug lead with that of the untreated starting material. All of the multicomponent forms showed an improvement in the maximum concentrations (Cmax) attained by the drug lead and the rate at which it dissolved. The recorded Cmax values exceeded the concentration of the untreated compound by factors in the range 4.6-5.6.
  • d-Glucamine:+Crystal+Structures+and+Dissolution+Study&rft.title=Journal+of+Pharmaceutical+Sciences&rft.issn=0022-3549&">Multicomponent Crystal of Mefenamic Acid and N-Methyl-d-Glucamine: Crystal
           Structures and Dissolution Study
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Erizal Zaini, Lili Fitriani, Risda Yulia Sari, Henni Rosaini, Ayano Horikawa, Hidehiro Uekusa A novel multicomponent crystal (MC) of mefenamic acid (MA) and N-methyl-d-glucamine (MG) had been prepared to improve the physicochemical properties of poorly soluble drugs, and was characterized for its physicochemical properties by powder X-ray diffraction analysis, differential scanning calorimetry thermal analysis, FT-IR spectroscopy, in vitro dissolution rate, and physical stability. In addition, the crystal structure was determined by single-crystal X-ray diffraction analysis. The differential scanning calorimetry thermogram of the MA-MG binary system exhibits a single and sharp endothermic peak at 151.20°C, which was attributed to the melting point of a MC of MA-MG. FT-IR spectroscopy analysis showed the occurrence of solid-state interaction by involving proton transfer between MA and MG. The crystal structure analysis confirmed that MA-MG formed 1:1 ratio salt type MC. The formation of a MC of MA with MG significantly improved the dissolution rate of MA in compared to intact MA, and also the crystal demonstrated a good stability under a high relative humidity. These good properties would be attributed to the layer structure of MA and MG in the crystal.
  • Simultaneous Analysis of Dissolution and Permeation Profiles of Nanosized
           and Microsized Formulations of Indomethacin Using the In Vitro
           Dissolution Absorption System 2
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Jibin Li, Li-Bin Li, Nourdine Nessah, Yuehua Huang, Carlos Hidalgo, Albert Owen, Ismael J. Hidalgo The in vitro dissolution absorption system 2 (IDAS2), a recent invention comprised a conventional dissolution vessel containing 2 permeation chambers with Caco-2 cell monolayers mounted with their apical side facing the dissolution media, permits simultaneous measurement of dissolution and permeation of drugs from intact clinical dosage forms. The objectives of this study were (1) to assess the utility of IDAS2 in the determination of the effect of particle size on in vitro performance of indomethacin and (2) to find out whether the behavior in IDAS2 of 2 indomethacin products differing in particle size is correlated with their in vivo behavior. Indomethacin dissolution and permeation across Caco-2 cell monolayers were simultaneously measured in IDAS2; the dissolution and permeation profiles were simultaneously modeled using a simple two-compartment model. Compared to microsized indomethacin, the nanosized formulation increased the dissolution rate constant by fivefold, whereas moderately increasing the permeation rate constant and the kinetic solubility. As a result, the drug amount permeated across the Caco-2 cell monolayers doubled in the nanosized versus microsized formulation. The in vitro results showed a good correlation with in vivo human oral pharmacokinetic parameters, thus emphasizing the physiological relevance of IDAS2 data in predicting in vivo absorption.
  • Identification of IgG1 Aggregation Initiation Region by Hydrogen Deuterium
           Mass Spectrometry
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Masanori Noda, Kentaro Ishii, Mika Yamauchi, Hiroaki Oyama, Takashi Tadokoro, Katsumi Maenaka, Tetsuo Torisu, Susumu Uchiyama Antibody aggregates are a potential risk for immunogenicity; therefore, rational approaches to improve associated aggregation properties need to be developed. Here, we report the amino acid region responsible for aggregation initiation. Two types of therapeutic IgG1 antibody monomer samples were prepared: IgG1 mAb40-3M stored at 40°C for 3 months, which existed in monodisperse state, and the monomer mAb65-5m, which was dissociated from small soluble aggregates by heating at 65°C for 5 min. Hydrogen deuterium exchange mass spectrometry of mAb40-3M identified 2 sites in the Fc region (site 1, F239-M256; site 2, S428-G450) with increased exchange rates. Site 1 includes a region reported as being susceptible to structural change induced by stress. Exposure of site 1 was undetected after 2 months of storage at 40°C but was subsequently detectable after 3 months. As site 2 is spatially close to site 1, the structural change of site 1 could propagate site 2. Besides these 2 regions, hydrogen deuterium exchange mass spectrometry of mAb65-5m identified an exposure of I257-W281 in Fc (site 3), within which a peptide sequence with high aggregation tendency was discovered. We thus concluded that exposure of site 3 is a trigger for the association of a partially denatured antibody.
  • Development of a Stable Liquid Formulation for Live Attenuated Influenza
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Yao Sun, Zhenwei Shen, Chun Zhang, Yanming Yi, Kunying Zhu, Fei Xu, Wei Kong Live attenuated influenza vaccine (LAIV) is considered one of the most effective vaccines and can be manufactured quickly and inexpensively to counter seasonal or pandemic influenza. Lyophilization is widely used in vaccine production. However, it requires a longer production cycle and large-scale equipment, thus posing a considerable financial burden for developing countries. A potential solution is the development of liquid LAIV, which can increase the yield and reduce the cost of production. In this study, influential factors of LAIV, such as potential stabilizing excipients and pH, were optimized by an orthogonal design. We found that pH is the most critical factor for the stability of LAIV; salt concentration and initial virus titer are also important for LAIV stability. With these data, we developed a liquid formulation consisting of 2.5% sucrose, 0.1% monosodium glutamate, 1% arginine, and 0.5% human serum albumin, with pH ranging from 6.2 to 6.9 (optimum pH 6.5-6.7), for optimal production of monovalent or trivalent LAIVs. This liquid formulation has the potential to considerably improve vaccine production capacity to compensate for the immense shortfall in influenza vaccines globally.
  • Temperature Measurement by Sublimation Rate as a Process Analytical
           Technology Tool in Lyophilization
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Hidenori Kawasaki, Toshinori Shimanouchi, Hiroyuki Sawada, Hiroshi Hosomi, Yuta Hamabe, Yukitaka Kimura Product temperature (Tb) and drying time constitute critical material attributes and process parameters in the lyophilization process and especially during the primary drying stage. In the study, we performed a temperature measurement by the sublimation rate (TMbySR) to monitor the Tb value and determine the end point of primary drying. First, the water vapor transfer resistance coefficient through the main pipe from the chamber to the condenser (Cr) was estimated via the water sublimation test. The use of Cr value made it possible to obtain the time course of Tb from the measurement of pressure at the drying chamber and at the condenser. Second, a Flomoxef sodium bulk solution was lyophilized by using the TMbySR system. The outcome was satisfactory when compared with that obtained via conventional sensors. The same was applicable for the determination of the end point of primary drying. A laboratory-scale application of the TMbySR system was evidenced via the experiment using 220-, 440-, and 660-vial scales of lyophilization. The outcome was not dependent on the loading amount. Thus, the results confirmed that the TMbySR system is a promising tool in laboratory scale.
  • Preservative and Irritant Capacity of Biosurfactants From Different
           Sources: A Comparative Study
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Lorena Rodríguez-López, Myriam Rincón-Fontán, Xanel Vecino, José Manuel Cruz, Ana Belén Moldes One of the most important challenges for pharmaceutical and cosmetic industries is solubilization and preservation of their active ingredients. Therefore, most of these formulations contain irritant chemical additives to improve their shelf-life and the solubility of hydrophobic ingredients. An interesting alternative to chemical surfactants and preservatives is the use of biosurfactants; thus, their surfactant properties and composition make them more biocompatible than their chemical counterparts. Moreover, some biosurfactants have shown antimicrobial activity in addition to their detergent capacity. In this work, the antimicrobial and irritant effect of 2 biosurfactant extracts was studied: one produced in a controlled fermentation process with Lactobacillus pentosus and the other produced from corn stream by spontaneous fermentation. The results showed a strong antimicrobial activity of the biosurfactant extract obtained from corn stream on pathogenic bacteria, in comparison with the L. pentosus biosurfactant extract. Moreover, both biosurfactants did not produce any irritant effect on the chorioallantoic membrane of hen's egg assay contrary to sodium dodecyl sulfate. This is the first study dealing with the application of biosurfactant extracts on sensitive biological membranes, and this is the first time that the preservative capacity of a biosurfactant extract obtained in spontaneous fermentation is being evaluated, achieving promising results.
  • Quality by Design-Driven Process Development of Cell Culture in Bioreactor
           for the Production of Foot-And-Mouth Veterinary Vaccine
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Xinran Li, Xuerong Liu, Rongbin Wang, Fanglan An, Jianqi Nie, Yunde Zhang, Hadji Ahamada, Xiuxia Liu, Chunli Liu, Yu Deng, Yankun Yang, Zhonghu Bai Quality by design (QbD) principle has been established as a guideline to emphasize the understanding of the relationship of product quality with process control. Vaccine product have characteristics of security and high efficiency, but it also has features such as complexity and rigorous regulatory for production. This case study describes an example of QbD-driven process development for manufacturing a veterinary vaccine produced with baby hamster kidney-21 cells. The study revealed that cell culture duration was the most significant factor affecting 50% tissue culture infectious doses (TCID50) and antigenic titer, and the factors of culture temperature and pH at infection phase exhibited less effect. Culture temperature at infection phase was the only significant factor for total protein. Through the Monte Carlo simulation, the design spaces of process parameters were determined. Meanwhile, the excellent and robust performance in manufacturing scale (4000-L) validated the effectiveness of this strategy. A reliable and robust multivariate process parameter range, that is, design space, was identified by this systematic approach. Our investigation presents a successful case of QbD principle, which encourages other researchers to combine the methodology into other biopharmaceutical manufacturing process.
  • Increased Expression of Renal Drug Transporters in a Mouse Model of
           Familial Alzheimer's Disease
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Yijun Pan, Kotaro Omori, Izna Ali, Masanori Tachikawa, Tetsuya Terasaki, Kim L.R. Brouwer, Joseph A. Nicolazzo It is well established that the expression and function of drug transporters at the blood-brain barrier are altered in Alzheimer's disease (AD). However, we recently demonstrated in a mouse model of AD that the expression of key drug transporters and metabolizing enzymes was modified in peripheral organs, such as the small intestine and liver, suggesting that systemic drug absorption may be altered in AD. The purpose of this study was to determine whether the expression of drug transporters in the kidneys differed between 8- to 9-month-old wild-type mice and APPswe/PSEN1dE9 (APP/PS1) transgenic mice, a mouse model of familial AD, using a quantitative targeted absolute proteomics approach. The protein expression of the drug transporters—multidrug resistance-associated protein 2, organic anion transporter 3, and organic cation transporter 2—was upregulated 1.6-, 1.3-, and 1.4-fold, respectively, in kidneys from APP/PS1 mice relative to wild-type mice. These results suggest that in addition to modified oral absorption of certain drugs, it is possible that the renal excretion of drugs that are multidrug resistance-associated protein 2, organic anion transporter 3, and organic cation transporter 2 substrates could be altered in AD. These changes could affect the interpretation of studies conducted during drug development using this mouse model of AD and potentially impact dosage regimens of such drugs prescribed in this patient population.
  • Developability Assessment of Physicochemical Properties and Stability
           Profiles of HIV-1 BG505 SOSIP.664 and BG505 SOSIP.v4.1-GT1.1 gp140
           Envelope Glycoprotein Trimers as Candidate Vaccine Antigens
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Neal Whitaker, John M. Hickey, Kawaljit Kaur, Jian Xiong, Nishant Sawant, Albert Cupo, Wen-Hsin Lee, Gabriel Ozorowski, Max Medina-Ramírez, Andrew B. Ward, Rogier W. Sanders, John P. Moore, Sangeeta B. Joshi, David B. Volkin, Antu K. Dey The induction of broadly neutralizing antibodies (bNAbs) is a major goal in the development of an effective vaccine against HIV-1. A soluble, trimeric, germline (gI) bNAb-targeting variant of the HIV-1 envelope glycoprotein (termed BG505 SOSIP.v4.1-GT1.1 gp140, abbreviated to GT1.1) has recently been developed. Here, we have compared this new immunogen with the parental trimer from which it was derived, BG505 SOSIP.664 gp140. We used a comprehensive suite of biochemical and biophysical methods to determine physicochemical similarities and differences between the 2 trimers, and thereby assessed whether additional formulation development efforts were needed for the GT1.1 vaccine candidate. The overall higher order structure and oligomeric states of the 2 vaccine antigens were quite similar, as were their thermal, chemical, and colloidal stability profiles, as evaluated during accelerated stability studies. Overall, we conclude that the primary sequence changes made to create the gl bNAb-targeting GT1.1 trimer did not detrimentally affect its physicochemical properties or stability profiles from a pharmaceutical perspective. This developability assessment of the BG505 GT1.1 vaccine antigen supports using the formulation and storage conditions previously identified for the parental SOSIP.664 trimer and enables the development of GT1.1 for phase I clinical studies.
  • An Integrated Analysis of Solid Form Change Impact on Solubility and
           Permeability: Case Study of Oral Exposure in Rats of an RAR Related Orphan
           Receptor C Inhibitor
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Po-Chang Chiang, Karthik Nagapudi, Jia Liu, James J. Crawford, Jason R. Zbieg, Emile Plise, Yuzhong Deng It is well acknowledged that the oral absorption of a drug can be influenced by its solubility, which is usually associated with its solid form properties. G1032 is a retinoic acid–related orphan receptor inverse agonist. Crystalline solid (form A) was identified with an aqueous solubility of 130 μg/mL. This form was used in an oral dose escalation study in rodents up to 300 mg/kg and achieved good exposures. Later on, a more stable crystalline hydrate (form B) was identified and the aqueous solubility was reduced to 55 μg/mL. A modeling exercise suggested that this solubility change would cause a 2-fold decrease in exposure at tested doses; however, the actual reduction was far larger than the model predicted. At high dose, exposure was found to be reduced by almost 10-fold. A parameter sensitivity analysis suggested that such a drop in exposure could be associated with permeability reduction as well. More in vitro permeability experiments were performed, indicating G1032 was an efflux transporter substrate. This finding was integrated into the modeling and the design for in vivo studies. Data obtained from those studies allowed us to better understand the causes of the higher-than-expected exposure change and enabled decision-making.
  • The Effects of Clioquinol on P-glycoprotein Expression and Biometal
           Distribution in the Mouse Brain Microvasculature
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Mitchell P. McInerney, Yijun Pan, Irene Volitakis, Ashley I. Bush, Jennifer L. Short, Joseph A. Nicolazzo Previous studies have demonstrated that the ionophore clioquinol (CQ), in conjunction with the biometals copper and zinc, increases the expression of P-glycoprotein (P-gp) in human cerebral microvascular endothelial (hCMEC/D3) cells. As P-gp expression and function at the blood-brain barrier (BBB) is of great interest regarding CNS drug access and endogenous toxin trafficking (e.g., amyloid beta), the present study assessed the in vivo translation of these previous in vitro findings. Swiss outbred mice received an 11-day treatment of CQ (30 mg/kg) by oral gavage, after which brain microvessel-enriched fractions (MEFs) and surrounding interfaces (subcortical brain tissue and plasma) were extracted. P-gp expression was quantified in the MEF, and biometal concentrations in all 3 compartments were assessed via inductively coupled plasma mass spectrometry. CQ treatment did not modify the expression of P-gp, nor copper or zinc concentrations in the brain MEF under this treatment regime. Metallomic analysis revealed, however, that CQ reduced potassium and magnesium levels in the brain MEF and also lowered brain iron levels. This study has shown that under this dosing regimen, CQ does not increase BBB P-gp expression in Swiss outbred mice, but that CQ facilitates redistribution of certain metal ions within the brain MEF, plasma, and brain parenchyma.
  • DNA Quadruplex–Based Inhibitor With Flexible Fragments at the
           3′ Terminal Shows Enhanced Anti–HIV-1 Fusion Activity
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Zeye Han, Yongjia Tang, Hongqian Ren, Keliang Liu, Liang Xu Chemically optimizing the molecular structure of aptamers may enhance properties such as biological activity or metabolic stability. DNA quadruplex–based HIV-1 fusion inhibitors were found to interact with HIV-1 surface glycoprotein in aptamer mode. In this work, a series of quadruplex-based HIV-1 fusion inhibitors with flexible oligodeoxynucleotide fragments at the 3′ terminal was discovered. The flexible extension did not greatly influence quadruplex formation at the 5′-end. Increasing the length of the flexible fragment may increase antifusion activity. Compared with a traditional inhibitor, d(5′TGGGAG3′)4, these novel inhibitors showed enhanced interaction with HIV-1 glycoproteins gp120 and gp41, which increased inhibition of 6-helical bundle formation during the course of virus fusion. These inhibitors also showed improved stability, compared with natural oligodeoxynucleotide. This work may inform the design of anti–HIV-1 DNA helix-based inhibitors with new structures or mechanisms.
  • Porous Polymeric Microparticles as an Oral Drug Platform for Effective
           Ulcerative Colitis Treatment
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Xin Zhou, Qiubing Chen, Ya Ma, Yamei Huang, Shuangquan Gou, Bo Xiao Porous microparticles (MPs) have been regarded as a promising vehicle for drug delivery. Herein, porous MPs and their counterparts (nonporous MPs) were produced by a conventional emulsion-solvent evaporation method in the presence and absence of ammonium bicarbonate, and curcumin was encapsulated into these MPs during the preparation process. The obtained MPs possessed desirable diameters of around 1.2 μm and negative zeta potentials of approximately −28 mV. It was found that the release rate of curcumin was remarkably increased with the introduction of pores in MPs. Furthermore, orally administered porous MPs achieved statistically significantly better therapeutic outcomes against ulcerative colitis mouse model induced by dextran sulfate sodium, in comparison to nonporous MPs. These findings confirmed that porous MPs could be served as a promising platform for the treatment of ulcerative colitis via oral route.
  • Innovation in Chemistry, Manufacturing, and Controls—A Regulatory
           Perspective From Industry
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Nina S. Cauchon, Shirley Oghamian, Soraya Hassanpour, Michael Abernathy This review describes the landscape of novel modalities such as cell and gene therapies, viruses, other novel biologics, oligomers, and emerging technologies, including modern analytics. We summarize the regulatory history and recent landmark developments in some major markets and examine specific chemistry, manufacturing, and controls (CMC) challenges, including suggestions for exploration of potential science-based approaches in support of regulatory strategy development from an industry perspective. In addition, we evaluate the economic factors contributing to patient access to innovation and discuss the impact of regulation. There is a desperate need for a consistent form of regulation where global approaches to regulatory strategies can be harmonized, and specific CMC challenges can be dealt with using the appropriate science and risk-based tools. Although these tools are well described in current guidance documents, the specifics of applicability to complex novel modalities can still result in differing regulatory advice and outcomes. The future goals for efficiently regulating innovative modalities and technologies could be aided by more regulatory harmonization, regulatory education, and industry cooperation through consortia, enabling industry to supply key information to regulators in a transparent yet well-defined manner, and utilizing mutually understood risk-benefit analyses to produce drugs with appropriate safety, efficacy, and quality characteristics.
  • Editorial Advisory Board
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s):
  • A Novel Pharmacokinetic Bridging Strategy to Support a Change in the Route
           of Administration for Biologics
    • Abstract: Publication date: July 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 7Author(s): Ping Ji, Nikolay Nikolov, Sally Seymour, Rachel Glaser, Yupeng Ren, Liang Li, Anshu Marathe, Yunzhao Ren, Jianmeng Chen, Lei He, Dipak Pisal, Shalini Wickramaratne Senarath Yapa, Yaning Wang, Chandrahas Sahajwalla Determination of appropriate pharmacokinetic end point to bridge different dosing regimens is often a challenge when developing a new route of administration. Trough concentrations (Ctrough) are often considered the most relevant PK end point to predict efficacy (ACR20/DAS28) in the treatment of rheumatoid arthritis for biologics. However, no systematic research has been conducted to evaluate this approach. We developed a novel strategy to predict the most relevant PK variables that may be used to support a change in the route of administration for biological products. Our analysis indicated that matching only Ctrough when switching from intravenous dosing to subcutaneous dosing with decreasing dosing interval may result in a lower treatment response. If only average concentration (Cave) is considered as the relevant variable, our analysis showed that treatment response may be worsened when switching from subcutaneous dosing to intravenous dosing with increasing dosing interval. The study results demonstrated that matching a single pharmacokinetic end point (Ctrough or Cave) may not be sufficient to ensure efficacy when switching between intravenous dosing and subcutaneous dosing. A practical novel pharmacokinetic bridging approach is provided to support a change in the route of administration for biological products.
  • DEHP Nanodroplets Leached from Polyvinyl Chloride IV Bags Promote
           Aggregation of IVIG and Activate Complement in Human Serum
    • Abstract: Publication date: Available online 21 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Jared R. Snell, Connor R. Monticello, Cheng Her, Emma L. Ross, Ashley A. Frazer-Abel, John F. Carpenter, Theodore W. Randolph Concerns regarding the impact of sub-visible particulate impurities on the safety and efficacy of therapeutic protein products have led manufacturers to implement strategies to minimize protein aggregation and particle formation during manufacturing, storage and shipping. However, once these products are released, manufacturers have limited control over product handling. In this work we investigated the effect of di(2-ethylhexyl) phthalate (DEHP) nanodroplets generated in polyvinyl chloride (PVC) bags of intravenous (IV) saline on the stability and immunogenicity of intravenous immunoglobulin (IVIG) formulations. We showed that PVC IV bags containing saline can release DEHP droplets into the solution when agitated or transported using a pneumatic tube transportation system in a clinical setting. We next investigated the effects of emulsified DEHP nanodroplets on IVIG stability and immunogenicity. IVIG adsorbed strongly to DEHP nanodroplets, forming a monolayer. In addition, DEHP nanodroplets accelerated IVIG aggregation in agitated samples. The immunogenicity of DEHP nanodroplets and IVIG aggregates generated in these formulations were evaluated using an in-vitro assay of complement activation in human serum. The results suggested DEHP nanodroplets shed from PVC IV bags could reduce protein stability and induce activation of the complement system, potentially contributing to adverse immune responses during the administration of therapeutic proteins.
  • Synergistic enhancement of cellular uptake with CD44-expressing malignant
           pleural mesothelioma by combining cationic liposome and hyaluronic
           acid-lipid conjugate
    • Abstract: Publication date: Available online 21 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Yu Sakurai, Akari Kato, Yasuhiro Hida, Junichi Hamada, Nako Maishi, Kyoko Hida, Hideyoshi Harashima Malignant pleural mesothelioma (MPM) is a highly aggressive form of cancer, with a median survival of less than one year. It is well known that the hyaluronan (HA) receptor CD44 is highly expressed by MPM cells and is reported to be correlated with a poor prognosis. We herein report on the development of a new type if drug delivery system against CD44 that involves the use of lipid nanoparticles (LNPs) equipped with a new type of HA derivative. In this study, we evaluated HA-lipid conjugation (HAL) via the end of the HA molecule through reductive amination, a process that allowed the carboxylate group to remain intact. As a result, the HAL-modified LNP appears to be a potent nanoparticle for dealing with MPM. Surprisingly, the use of a combination of a cationic lipid and HAL had a synergistic effect on cellular uptake in MPM and consequently permitted an anti-cancer drug such as cis-diamminedichloro-platinum(II) (CDDP). Intrapleural injection of CDDP-loaded HAL-LNP (1.5 mg/kg as CDDP) per week significantly suppressed the progression of this type of cancer in a MPM orthotopic model. These results suggest that HAL-modified LNP represents a potent delivery system for MPM cells that express high levels of CD44.
  • Solubilization of Cyclosporine in Topical Ophthalmic Formulations:
           Preformulation Risk Assessment on A New Solid Form
    • Abstract: Publication date: Available online 20 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Ke Wu, Anu Gore, Richard Graham, Richard Meller Due to the discovery of a less soluble crystalline form (Form 2) of cyclosporine (CsA), risks in solubility and physical stability of these formulations need to be revisited. This work focused on understanding the solubility behavior of various CsA forms in different media, including water, castor oil, and selected cosolvent micellar systems. In water, Form 2 was approximately 8-9 times less soluble than Form 1 (aka. tetragonal dihydrate). In neat non-aqueous solvent, e.g., castor oil, Form 3 (aka. orthorhombic hydrate) was found to have the lowest solubility and therefore the most stable form. Also, the solubility-temperature relationship of CsA is complex and solvent-dependent. In aqueous vehicles, retrograde temperature dependence of solubility was observed in aqueous vehicles, i.e., the solubility of CsA decreased with temperature, which was attributed to the effect of temperature on the strength of hydrogen bonding interactions; conversely, the solubility of CsA increased with temperature in non-aqueous solvents. In addition, the solubility of these CsA forms was very sensitive to temperature. Temperature-dependent form transformation was also observed in the media studied, with faster form conversion occurring at elevated temperatures. These studies provided key information to support the risk assessment for topical ophthalmic formulation development of CsA.
  • Identification of an adduct impurity of an active pharmaceutical
           ingredient and a leachable in an ophthalmic drug product using LC-QTOF
    • Abstract: Publication date: Available online 18 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Ramarao Gollapalli, Gagandeep Singh, Alejandro Blinder, Jeremiah Brittin, Arijit Sengupta, Bikash Mondal, Milan Patel, Biswajit Pati, James Lee, Amit Ghode, Mahesh Kote Impurity investigations are important in pharmaceutical development to ensure drug purity and safety for the patient. The impurities typically found in drug products are degradants or reaction products of the active pharmaceutical ingredient (API), or leachable compounds from the container closure system. However, secondary reactions may also occur between API degradants, excipient impurities, residual solvents and leachables to form adduct impurities. We hereby report an adduct-forming interaction of API (Moxifloxacin) with a leachable compound (ethylene glycol monoformate) in Moxifloxacin ophthalmic solution. The leachable compound originated from a low density polyethylene (LDPE) bottle employed in the packaging of drug products. The adduct impurity was tentatively identified as 1-cyclopropyl-6-fluoro-7-(1-(2-(formyloxy)ethyl)octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl)-8-methoxy-4-oxo-1,4-dihydroquinoline-3-carboxylic acid (C24H28FN3O6, MW= 473.19621) using accurate mass LC-QTOF analysis. The mass accuracy error between the theoretical mass and the experimental mass of an impurity was found to be 0.2 ppm. An MS/MS analysis was utilized to provide mass spectrometry fragments to support verification of the proposed structure.
  • De-risking early stage drug development with a bespoke lattice energy
           predictive model: A materials science informatics approach to address
           challenges associated with a diverse chemical space.
    • Abstract: Publication date: Available online 18 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Ernest Chow, Robert Docherty, Tiffany Lai, Klimentina Pencheva The solid-state properties of new chemical entities are critical to the stability and bioavailability of pharmaceutical drug products. The stability of the solid-state packing is described by the packing energy and an accurate prediction of this property for drug molecules would therefore be desirable. However, this has been difficult to achieve because of the lack of fundamental thermodynamic data on drug molecules. A potential solution would be to use calculated lattice energies to build a model and design molecules with desired physicochemical properties from an early stage, aligning with a ‘design by first intent’ strategy for physicochemical properties. We first demonstrate the high correlation and interchangeability between QSPR models built using calculated lattice energies and experimental sublimation enthalpies for small organic molecules. We then present a QSPR model trained on in-house molecules using lattice energies calculated from crystal structures. The result is a model that enables fast prediction of the lattice energies of in-house molecules from 2-D molecular structure with reasonable accuracy (R2 = 0.92, RMSE = 3.58 kcal/mol). We explore the model elements to improve our understanding of the molecular properties that contribute to lattice energy, then suggest potential cross-industry aspects that may enhance the application of the concept.
  • In vitro and in vivo evaluation of core–shell mesoporous silica as a
           promising water-insoluble drug delivery system: Improving the dissolution
           rate and bioavailability of celecoxib with needle-like crystallinity
    • Abstract: Publication date: Available online 18 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Cuiyan Han, Shanqiang Zhang, Haitao Huang, Yan Dong, Xiaoyu Sui, Baiyu Jian, Wenquan Zhu The objective of our study was to prepare mesoporous silica nanoparticles with a core–shell structure (CSMSNs) and improve the dissolution and bioavailability of Celecoxib (Cxb), a water-insoluble drug, by changing its needlelike crystal form. CSMSNs are prepared by a core-shell segmentation self-assembly method. The SBET and Vt of CSMSNs were 890.65 m2/g and 1.23 cm3/g, respectively. Cxb was incorporated into CSMSNs by the solvent evaporation method. The gastrointestinal irritancy of the CSMSNs was evaluated by a gastric mucosa irritation test. In vitro dissolution and in vivo pharmacokinetic tests were carried out to study the improvement in the dissolution behavior and oral bioavailability of Cxb. In conclusion, Gastric mucosa irritation study indicated the good biocompatibility of CSMSNs. The cumulative dissolution of CSMSNs-Cxb is 86.2% within 60 min in SIF solution, which may be ascribed to the crystal form change caused by control of the nano-channel for CSMSNs. Moreover, CSMSNs could enhance the 9.9-fold AUC of Cxb. The cumulative dissolution and bioavailability of Cxb were both significantly enhanced by CSMSNs. CSMSNs with a core–shell structure are suitable as a carrier for a poorly water-soluble drug (Cxb).
  • Aggregate forms of recombinant human erythropoietin with different charge
           profile substantially impact biological activities
    • Abstract: Publication date: Available online 16 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Mansoureh Ghezlou, Farzad Mokhtari, Alireza Kalbasi, Gholamhossein Riazi, Hooman Kaghazian, Reza Emadi, Amir Reza Aref Recombinant human erythropoietin (EPO) as a glycoprotein growth factor has been considered a biological drug for treatment of anemic patients with chronic renal failure or who receive cancer chemotherapy. Biological activity and circulation time are two parameters are important to achieve EPO’s efficacy. Previous efforts for increasing EPO’s efficacy have focused on glycosylation modification via adding more sialic acid antenna and generate more negative charged protein. Evidences cleared that EPO’s activity increased by numbers of N-glycan moieties with presence of sialic acids at their terminus. Correlation between bioactivity and glycosylation with terminal sialylation is theoretically achieved using the calculation of the amount of charge profile of the EPO variants called “I-number”.Here we studied and compared the relationship between bioactivities of different EPOs that contained various I-numbers and the effect of their secondary and tertiary protein structures on measured in vivo efficacy. Eight recombinant EPOs batches were produced under the same condition. I-numbers found out by EPO’s charge profiles determination using capillary electrophoresis and activities were studied upon erythroid precursor cell stimulation in mice. Analyzing the bioactivity, I-number and structural studies revealed that in spite of I-number, conformational changes in protein structure and presence of aggregated species impact bioactivity substantially.
  • Developing cream formulations: renewed interest in an old problem
    • Abstract: Publication date: Available online 16 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Ana Simões, Francisco Veiga, Carla Vitorino This work aimed at establishing a framework to screen and understand the product variability deeming from factors that affect the quality features of cream formulations.As per Quality by Design (QbD)-based approach, cream quality target profile (QTPP) and critical quality attributes (CQAs) were identified, and a risk assessment analysis was conducted to qualitatively detect the most critical variables for cream design and development. A Plackett-Burman design was used to screen out unimportant factors, avoiding collecting large amounts of data. Accordingly, two designs of experiments (DoE-1 and DoE-2) were performed and the effects of independent variables on the cream formulations responses estimated. At different factor combinations, significant variability was observed in droplet size, consistency, hardness, compressibility and adhesiveness with values ranging from 2.60±0.88 to 9.91±5.63 μm, 7.93±0.05 to 13.53±0.14 mm, 27.60±0.30 to 58.39±1.10 g, 37.98±6.20 to 447.18±37.20 g.s and 25.66±2.12 to 286.30±32.86 g.s, respectively. The statistical analysis allowed determining the most influent factors. This study revealed the potential of QbD methodology in understanding product variability, recognizing the most critical independent variables for the final product quality. This systematic approach in the pharmaceutical field will yield more robust products and processes, provisioning time and cost effective developments.
  • Effect of iron oxide nanoparticles on the oxidation and secondary
           structure of growth hormone
    • Abstract: Publication date: Available online 16 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Ninad Varkhede, Bjo¨rn-Hendrik Peters, Yangjie Wei, C. Russell Middaugh, Christian Schöneich, M. Laird Forrest Oxidation of therapeutic proteins (TPs) can lead to changes in their pharmacokinetics, biological activity and immunogenicity. Metal impurities such as iron are known to increase oxidation of TPs, but nanoparticulate metals have unique physical and chemical properties compared to the bulk material or free metal ions. Iron oxide nanoparticles (IONPs) may originate from equipment used in the manufacturing of TPs or from needles during injection. In this study, the impact of IONPs on oxidation of a model protein, rat growth hormone (rGH), was investigated under chemical stress. Hydrogen peroxide (H2O2)- and 2,2′-azobis (2-methylpropionamidine) dihydrochloride (AAPH) oxidized methionine residues of rGH, but unexpectedly, oxidation was suppressed in the presence of IONPs compared to a phosphate buffer control. Fourier transform infrared (FTIR) spectroscopy indicated splitting of the α-helical absorbance band in the presence of IONPs, while CD spectra showed a reduced α-helical contribution with increasing temperature for both rGH and rGH-IONP mixtures. The results collectively indicate that IONPs can increase the chemical stability of rGH by altering the kinetics and preference of amino acid residues that are oxidized, although the changes in protein secondary structure by IONPs may lead to alterations of physical stability.
  • Impact of Surfactant and Surfactant-Polymer Interaction on
           Desupersaturation of Clotrimazole
    • Abstract: Publication date: Available online 14 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Wei Zhang, Siddhi S. Hate, David J. Russell, Hao Helen Hou, Karthik Nagapudi The impact of surfactants on supersaturation of clotrimazole solutions was systematically evaluated. Four clinically-relevant surfactants, sodium dodecyl sulfate (SDS), vitamin E TPGS, Tween 80 and docusate sodium were studied. The induction time for nucleation and rate of desupersaturation were determined at a supersaturation ratio of 90% amorphous solubility. Measurement was also performed in the presence of pre-dissolved hydroxypropyl methylcellulose acetate succinate (HPMCAS) to study the effect of surfactant-polymer interaction on desupersaturation. The four surfactants showed varied effects on desupersaturation. From supersaturation maintenance perspective, in the presence of HPMCAS, the rank order for the four surfactants was found to be: docusate sodium> vitamin E TPGS> SDS> Tween 80. Given the importance of maintaining supersaturation and varied effect of surfactants on nucleation kinetics and desupersaturation rate, a careful examination of API, polymer and surfactant interaction on an individual basis is recommended for selecting an appropriate surfactant for use in amorphous solid dispersion (ASD) formulation.
  • Characterization of Excipient Effects on Reversible Self-association,
           Backbone Flexibility, and Solution Properties of an IgG1 Monoclonal
           Antibody at High Concentrations: Part 1
    • Abstract: Publication date: Available online 13 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Yue Hu, Jayant Arora, Sangeeta B. Joshi, Reza Esfandiary, C. Russell Middaugh, David D. Weis, David B. Volkin Many challenges limit the formulation of antibodies as high-concentration liquid dosage forms including elevated solution viscosity, decreased physical stability, and in some cases, liquid-liquid phase separation. In this work, an IgG1 monoclonal antibody (mAb-J), which undergoes concentration dependent reversible self-association (RSA), is characterized in the presence of four amino acids (Arg, Lys, Asp, Glu) and NaCl using biophysical techniques and hydrogen exchange-mass spectrometry (HX-MS). The five additives disrupt RSA, prevent phase separation, and reduce solution viscosity to varying extents. These excipients also cause decreased turbidity, reduced average hydrodynamic diameter, and increased relative solubility of mAb-J in solution. The RSA disrupting efficacy of the positively-charged amino acids is greater than either negatively-charged amino acids or NaCl. As measured by HX-MS, anionic excipients induced more alterations of mAb-J backbone dynamics at pH 6.0, and weak Fab-Fab interactions likely remained with the addition of either cationic or anionic excipients at high protein concentrations. Along with a companion paper examining a different mAb with a different molecular mechanism of RSA, these results are discussed in the context of various excipient strategies to disrupt protein-protein interactions to formulate mAbs at high protein concentrations with good stability profiles and favorable pharmaceutical properties for subcutaneous administration.
  • Forced degradation of monoclonal antibodies after compounding: impact on
           routine hospital quality control
    • Abstract: Publication date: Available online 13 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): E. Jaccoulet, T. Daniel, P. Prognon, E. Caudron Compounded therapeutic monoclonal antibodies (mabs) used in a hospital require quality control (QC). In our hospital, analytical QC process intended to mabs identification and quantification is based on flow injection analysis (FIA) associated with second derivative UV spectroscopy and matching method algorithm. We studied the influence of degraded mabs after compounding on this validated QC. Three forced stress conditions including mechanical, thermal and freeze-thawing stresses were studied to yield degraded mabs from two model compounds, i.e., bevacizumab (IgG1) and nivolumab (IgG4). Different degraded mabs were generated and were analyzed in terms of turbidity, the percentage of aggregation, size distribution, and changes in tertiary structure. Stresses showed to be mab-dependent in terms of aggregation. Tertiary structural changes were observed in most of the stressed samples by principal component analysis (PCA) of the UV second derivative data. The structural and physicochemical modifications conducted to mismatch depending on the nature of the stress. The mismatch ranged from 17 % to 72 % for the mabs, except for freeze-thawed bevacizumab for which a perfect match (100 %) was reached. The quantification with an unfulfilled relative error of the concentration (i.e.,> ± 15 %) was detected only for mechanically stressed mabs. In conclusion, the study revealed that the influence of the mabs and the type of stress impact on the QC of compounded mabs.
  • Nanostructured Lipid Carriers for oral bioavailability enhancement of
           Exemestane: Formulation design, in vitro, ex vivo and in vivo studies
    • Abstract: Publication date: Available online 13 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Archu Singh, Yub Raj Neupane, Bharti Mangla, Kanchan Kohli Exemestane (EXE), a novel oral steroidal aromatase inhibitor, approved for the treatment of breast cancer. However, its oral clinical application is limited because of low aqueous solubility and low oral bioavailability. Here, we aim to design and fabricate Nanostructured Lipid Carriers (NLCs) using Precirol® ATO5 and Flaxseed oil as the solid lipid and liquid lipid, respectively. EXE loaded NLCs were spherical in shape and with the hydrodynamic diameter of 131.3±2.43 nm, polydispersity index (PDI) 0.205 ± 0.06 and % Entrapment efficiency (% EE) 85.6 ± 1.20%. In vitro release study demonstrated a sustained release pattern for 24 h with relative burst release at the initial time point. Differential scanning calorimetry (DSC) and Powder X-ray diffraction (PXRD) studies showed reduced crystallinity and complete encapsulation of drug within lipid matrix. Ex vivo gut permeation study and Confocal Laser Scanning Microscopy (CLSM) revealed that NLCs comprising of lipid blend and surfactant enhanced intestinal permeability of EXE. Moreover, in vivo pharmacokinetic study on female Wistar rats found to 3.9-fold augment in oral bioavailability of EXE through NLCs compared with EXE suspension. Herein, we depict that loading of EXE into NLCs hold promising approach for the oral delivery of EXE in cancer therapy.
  • Hybrid nanostructured films for topical administration of simvastatin as
           coadjuvant treatment of melanoma
    • Abstract: Publication date: Available online 12 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Antonella Barone, Maria Mendes, Célia Cabral, Rosario Mare, Donatella Paolino, Carla Vitorino This work aims at (i) assessing the potential of repurposing simvastatin (SV) to support the most common therapies against melanoma, and (ii) developing an innovative topical adhesive film, composed by chitosan coated nanostructured lipid carriers (Ch-NLC) used as drug vehicle. A factorial design approach was employed as the basis for the formulation development. Optimized Ch-NLC displayed a particle size of 108±1 nm, a polydispersity index of 0.226, a zeta potential of 17.0±0.6 mV, as well as an entrapment efficiency of 99.86±0.08%, and SV loading of 14.99±0.01%. The performance of SV-Ch-NLC films was assessed in terms of release, permeation and adhesion, as critical quality attributes. Cutaneous tolerability and in vitro cytotoxicity studies were performed to warrant film safety and drug effectiveness, respectively.The topical films provided a sustained release kinetic profile of SV and were classified as non-irritant systems. The encapsulation of SV increased cytotoxicity in melanoma cells. The key role of squalene as nanostructuring agent of the lipid nanoparticle matrix, and as permeation enhancer was highlighted, suggesting its key action for potentiating skin permeation and uptake into melanoma cells. Topical SV-Ch-NLC films are thus, able to provide an in situ extended drug delivery, useful as coadjuvant treatment of melanoma skin lesions.
  • Characterization of Excipient Effects on Reversible Self-association,
           Backbone Flexibility, and Solution Properties of an IgG1 Monoclonal
           Antibody at High Concentrations: Part 2
    • Abstract: Publication date: Available online 11 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Yue Hu, Ronald T. Toth, Sangeeta B. Joshi, Reza Esfandiary, C. Russell Middaugh, David B. Volkin, David D. Weis In this work, we continue to examine excipient effects on the reversible self-association (RSA) of two different IgG1 monoclonal antibodies (mAb-J and mAb-C). We characterize the RSA behavior of mAb-C which, similar to mAb-J (see companion paper), undergoes concentration-dependent RSA, but by a different molecular mechanism. Five additives that affect protein hydrophobic interactions to varying extents including a chaotropic salt (guanidine hydrochloride, GdnHCl), a hydrophobic salt (trimethylphenylammonium iodide, TMPAI), an aromatic amino acid derivative (tryptophan amide hydrochloride, TrpNH2HCl), a kosmotropic salt (sodium sulfate, Na2SO4), and a less polar solvent (ethanol) were evaluated to determine their effects on the solution properties, molecular properties, and RSA of mAb-C at various protein concentrations. Four of the five additives examined demonstrated favorable effects on the pharmaceutical properties of high concentration mAb-C solutions (i.e., lower viscosity and weakened protein-protein interactions, PPIs) with a ranking order of GdnHCl> TMPAI> TrpNH2HCl> ethanol as measured by various biophysical techniques. Conversely, addition of Na2SO4 resulted in less desirable solution properties and enhanced PPIs. The effect of these five additives on mAb-C backbone dynamics were evaluated by HX-MS (at high vs. low protein concentrations) to better understand their effects on the molecular sites of RSA in mAb-C.
  • Biowaiver Monograph for Immediate-Release Solid Oral Dosage Forms:
    • Abstract: Publication date: Available online 8 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Gopal Singh Rajawat, Tejashree Belubbi, Mangal S. Nagarsenker, Bertil Abrahamsson, Rodrigo Cristofoletti, Dirk W. Groot, Peter Langguth, Alan Parr, James E. Polli, Mehul Mehta, Vinod P. Shah, Tomokazu Tajiri, Jennifer Dressman Literature data pertaining to the physicochemical, pharmaceutical and pharmacokinetic properties of ondansetron hydrochloride dihydrate are reviewed to arrive at a decision on whether a marketing authorization of an immediate release solid oral dosage form can be approved based on a BCS-based biowaiver. Ondansetron, a 5HT3 receptor antagonist, is used at doses ranging from 4mg to 24 mg in the management of nausea and vomiting associated with chemotherapy, radiotherapy and postoperative treatment. It is a weak base and thus exhibits pH dependent solubility. However, it is able to meet the criteria of ‘high solubility’ as well as ‘high permeability’ and can therefore be classified as a BCS class I drug. Further, ondansetron hydrochloride 8mg IR tablets (Zofran 8 mg) and multiples thereof (16 mg =Zofran 8mg × 2 tablets and 24 mg =Zofran 8mg × 3 tablets) meet the criteria of ‘rapidly dissolving’ in dissolution testing. Ondansetron hydrochloride has a wide therapeutic window and is well-tolerated after oral administration. Based on its favorable physicochemical properties, pharmacokinetic data and the minimal risks associated with an incorrect bioequivalence decision, the BCS-based biowaiver procedure can be recommended for ondansetron hydrochloride dihydrate immediate release tablets.
  • Design of virus-mimicking polyelectrolyte complexes for enhanced oral
           insulin delivery
    • Abstract: Publication date: Available online 6 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Chang Liu, Huan Xu, Ying Sun, Xin Zhang, Hongbo Cheng, Shirui Mao The viscous and elastic mucus layer is still an undesirable barrier for oral insulin delivery. To solve the problem, virus-mimicking nano-sized polyelectrolyte complex (PEC) was designed and their capacity in enhancing peroral insulin absorption in combination with bifunctional material SDS coating was investigated. Inspired by nature, virus-mimicking chitosan (CS)-modified L-Phe derivatives were synthesized to simulate the components of viral envelopes and then PECs between CS-g-N-Phe copolymers and insulin were prepared to achieve both structure and composition simulation of virus envelop. Based on the results from both in vitro and in vivo studies, it was concluded that in vitro mucodiffusion and in vivo hypoglycemic effect were dependent on L-Phe graft ratio, with CS-g-N-Phe20.2%/Insulin PECs presenting 2.0-2.2 fold higher relative pharmacological bioavailability than non-modified CS/Insulin PECs. Thereafter, SDS solution was applied as outer layer coating on the surface of virus-mimicking PECs. The coated PECs showed improved enzymatic stability, enhanced transport across mucus layer as well as intestinal epithelium in a SDS concentration dependent manner, with 0.6% SDS coating presenting the best effect, with further enhanced relative pharmacological bioavailability in healthy rats and prolonged therapeutic effect up to 9h.
  • Considerations for the Design of Antibody-Based Therapeutics
    • Abstract: Publication date: Available online 4 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Dennis R. Goulet, William M. Atkins Antibody-based proteins have become an important class of biologic therapeutics, due in large part to the stability, specificity, and adaptability of the antibody framework. Indeed, antibodies not only have the inherent ability to bind both antigens and endogenous immune receptors; they have also proven extremely amenable to protein engineering. Thus, several derivatives of the monoclonal antibody format, including bispecific antibodies, antibody-drug conjugates, and antibody fragments, have demonstrated efficacy for treating human disease, particularly in the fields of immunology and oncology. Reviewed here are considerations for the design of antibody-based therapeutics, including immunological context, therapeutic mechanisms, and engineering strategies. First, characteristics of antibodies are introduced, with emphasis on structural domains, functionally important receptors, isotypic and allotypic differences, and modifications such as glycosylation. Next, aspects of therapeutic antibody design are discussed, including identification of antigen-specific variable regions, choice of expression system, utility of multispecific formats, and design of antibody derivatives based on fragmentation, oligomerization, or conjugation to other functional moieties. Finally, strategies to enhance antibody function through protein engineering are reviewed while highlighting the impact of fundamental biophysical properties on protein developability.
  • Solid state reactivity of mechano-activated simvastatin: Atypical relation
           to powder crystallinity
    • Abstract: Publication date: Available online 4 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Dattatray Modhave, Peter Laggner, Michael Brunsteiner, Amrit Paudel The present study investigated the impact of solid state disorders generated during milling on the chemical reactivity of simvastatin. An amorphous and a partially crystalline simvastatin powders were generated via cryo-milling simvastatin crystals for either 90 or 10 min, respectively. The thoroughly characterized milled powders were stored at 40 °C/75% RH, in open and closed containers. The effect of milling and storage conditions on physical stability was investigated using simultaneous small and wide angle X-ray scattering and differential scanning calorimetry. The chemical degradation was evaluated using liquid chromatography-mass spectrometry. Compared to the fully amorphous state, the partially crystalline simvastatin crystallized to a lower extent in the expense of higher chemical degradation upon open storage. The closely stored samples degraded to a lower extent and crystallized to a higher extent than the openly stored ones. However, the trends of the total crystallinity and degradation between amorphous and partially crystalline powders were similar. Small angle X-ray scattering revealed that the partially crystalline simvastatin comprised a higher extent of nanoscale density heterogeneity than the fully amorphous powder. The overall results pointed towards the role of the remaining amorphous content and the nanoscale/ mesoscale density heterogeneity on the chemical reactivity in the disordered simvastatin.
  • Some Preformulation Studies of Pyruvic Acid and Other α-Keto Carboxylic
           Acids in Aqueous Solution: Pharmaceutical Formulation Implications for
           These Peroxide Scavengers
    • Abstract: Publication date: Available online 1 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Antonio Lopalco, Rodney Deeken, Justin Douglas, Nunzio Denora, Valentino J. Stella The purpose of this study is to assess some of the variables determining the aldol-like condensation of pyruvic acid (1), a peroxide scavenger, in aqueous solution to parapyruvic acid and higher oligomers. Its stability is compared to three other α-keto carboxylic acids, two with sterically hindered methylene groups alpha to the keto functionality (2-3) and phenylglyoxylic acid (4) with no methylene group. High performance liquid chromatography, nuclear magnetic resonance and liquid chromatography mass spectroscopy techniques are used to follow the kinetics and product analyses. 1 condensation is concentration dependent and base catalyzed above pH 7 consistent with the reaction mechanism proceeding through the attack of the fraction of the methylene group, alpha to the keto group, in its anionic form, at the keto group of a second molecule of 1. The major product is confirmed to be parapyruvic acid but higher order oligomers are also observed. All three of the other α-keto carboxylic acids 2-4 are considerably less reactive with 4 being completely stable. Stable solutions of 1 can be prepared by the use of relatively dilute solutions maintained at slightly acidic pH values. 1 prevents the oxidation of methionine on addition of hydrogen peroxide.
  • Nilotinib alters the efflux transporter-mediated pharmacokinetics of
           afatinib in mice
    • Abstract: Publication date: Available online 1 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Ronilda R. D’Cunha, Daryl J. Murry, Guohua An Small molecule tyrosine kinase inhibitors (TKIs) are novel anticancer agents with enhanced selectivity and superior safety profiles than conventional chemotherapeutics. A major shortcoming in TKI therapy is the development of acquired resistance. An important resistance mechanism is reduced intracellular drug accumulation due to an overexpression of efflux transporters such as P-glycoprotein (Pgp) and Breast Cancer Resistance Protein (BCRP) in cancer cells. TKIs have dual roles as substrates and inhibitors of Pgp and BCRP, thus combination TKI therapy could potentially reverse efflux transporter-mediated TKI resistance. In the current study, the effect of 14 TKIs on Pgp-, Bcrp1- and BCRP- mediated afatinib efflux was investigated in-vitro. Nilotinib was a potent inhibitor of Pgp, Bcrp1 and BCRP, with EC50 values of 2.22, 2.47 and 0.692μM, respectively. Consequently, the pharmacokinetics of afatinib with and without the co-administration of nilotinib was determined in mice plasma and various tissues. Nilotinib increased afatinib AUC by 188% in plasma and this altered tissue AUC by -38.8 to +221%. Nilotinib also decreased the clearance of afatinib by 65.3%, from 609 to 211mL/hr. Further studies are warranted to assess nilotinib’s chemosensitizing effect in tumor xenograft models.
  • Understanding the influence of nanocarrier-mediated brain delivery on
           therapeutic performance through pharmacokinetic-pharmacodynamic modeling
    • Abstract: Publication date: Available online 1 June 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Yang Hu, Margareta Hammarlund-Udenaes, Markus Fridén This study aimed at evaluating how encapsulation in a regular nanocarrier (NC) (providing extended circulation time), or in a brain-targeting NC (providing prolonged circulation time and increased brain uptake) may influence the therapeutic index compared to unformulated drug, and to explore the key parameters affecting therapeutic performance using a model-based approach. Pharmacokinetic (PK) models were built with chosen PK parameters. For a scenario where central effect depends on area under the unbound brain concentration curve and peripheral toxicity relates to peak unbound plasma concentration, dose-effect/side effect curves were constructed and the therapeutic index was evaluated. Regular NC improved the therapeutic index compared to unformulated drug due to reduced peripheral toxicity, while brain-targeting NC enhanced the therapeutic index by lowering peripheral toxicity and increasing central effect. Decreasing the drug release rate or systemic clearance of NC with drug still encapsulated could increase the therapeutic index. Also, a drug with shorter half-life would therapeutically benefit more from a NC encapsulation. This work provides insights into how a NC for brain delivery should be optimized to maximize the therapeutic performance, and is helpful to predict if and to what extent a drug with certain PK properties would obtain therapeutic benefit from nanoencapsulation.
  • Detection and Sizing of Submicron Particles in Biologics With
           Interferometric Scattering Microscopy
    • Abstract: Publication date: Available online 31 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Nathan A. Wong, Nina V. Uchida, Thilini U. Dissanayake, Mehulkumar Patel, Maira Iqbal, Taylor J. Woehl We demonstrate the application of interferometric scattering microscopy (IFS) for characterizing submicron particles in stir-stressed monoclonal antibody. IFS uses a layered silicon sensor and modified optical microscope to rapidly visualize and determine the particle size distribution (PSD) of submicron particles based on their scattering intensity, which is directly proportional to particle mass. Limits for particle size and optimal solution concentration were established for IFS characterization of submicron particles. We critically compare IFS data with dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA) and find IFS is superior to NTA and DLS for determining the realistic shape of the number-based PSD, whereas NTA and DLS provide superior information about absolute particle size. Together, IFS, NTA, and DLS provide complementary information on submicron particles and enable quantitative characterization of the PSD of submicron aggregates. Finally, we explore quantifying particle size with IFS by developing a calibration curve for particle scattering intensity based on correlative scanning electron microscopy imaging. We found that only a subset of isotropic-shaped particles followed the expected proportionality between IFS intensity and particle mass. Overall, this study demonstrates IFS is a simple approach for detecting and quantifying submicron aggregate PSD in protein-based therapeutics.
  • A Spray-dried combination of capreomycin and cpzen-45 for inhaled
           tuberculosis therapy
    • Abstract: Publication date: Available online 29 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Ragan A. Pitner, Phillip G. Durham, Ian E. Stewart, Steven G. Reed, Gail H. Cassell, Anthony J. Hickey, Darrick Carter Tuberculosis (TB) remains the single most serious infectious disease attributable to a single-causative organism. A variety of drugs have been evaluated for pulmonary delivery as dry powders: capreomycin sulfate has shown efficacy and was safely delivered by inhalation at high doses to human volunteers, while CPZEN-45 is a new drug that has also been shown to kill resistant TB. The studies here combine these drugs — acting by different mechanisms — as components of single particles by spray drying, yielding a new combination drug therapy. The spray dried combination powder was prepared in an aerodynamic particle size range suitable for pulmonary delivery. Physicochemical storage stability was demonstrated for a period of 6 months. The spray-dried combination powders of capreomycin and CPZEN-45 have only moderate affinity for mucin, indicating that delivered drug will not be bound by these mucins in the lung and available for microbicidal effects. The pharmacokinetics of disposition in guinea pigs demonstrated high local concentrations of drug following direct administration to the lungs and subsequent systemic bioavailability. Further studies are required to demonstrate the in vivo efficacy of the combination to confirm the therapeutic potential of this novel combination.
  • Translating Cell and Gene biopharmaceutical products for health and market
           impact Product scaling from clinical to marketplace: Lessons learned and
           future outlook
    • Abstract: Publication date: Available online 29 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Annette Bak, Kristina Pagh Friis, Yan Wu, Rodney JY. Ho Cell and gene therapies have the potential to be curative for severe disease states such as cancer or incurable orphan genetic diseases. Despite the promise there are only few such therapies available, although more are appearing in pharmaceutical pipelines. A major culprit limiting a fast translation from preclinical research to the clinic and the market is chemistry manufacturing and control (CMC). The root-course is that most cell and gene therapies currently are personalized in form of ex vivo manipulated cells. This approach stands in sharp contrast to the population-based approach seen for small molecules and protein therapeutics. Therefore, it warrants a different approach to product manufacturing, testing, release, regulatory submissions, and product distribution. In this commentary, we highlight opportunities to solve these issues already in progress in industry and at academic institutions, but in addition call for expert contributions to a future cluster of articles in Journal of Pharmaceutical Sciences to illuminate additional solutions. Finally, we are also providing a perspective on future directions including expanding from current approaches of gene modification via viral vectors to for example gene editing, approaches that may lend themselves better toward allogenic and in vivo therapies and more typical CMC approaches.
  • Optimization and application of in vitro and ex vivo models for vaginal
           semisolids safety evaluation
    • Abstract: Publication date: Available online 29 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Rita Monteiro Machado, Ana Palmeira-de-Oliveira, Luiza Breitenfeld, José Martinez-de-Oliveira, Rita Palmeira-de-Oliveira Preclinical safety assessment of vaginal products includes cytotoxicity assays upon cell lines. Furthermore, tissue explants have been considered for application on ex vivo models. In this study, traditional and renewed methods were studied for toxicity assessment of vaginal semisolids upon products currently used in clinical practice as antimicrobials (Gino-Canesten®, Sertopic®, Dermofix®, Gyno-pevaryl®, Lomexin®, Gino Travogen®, Dalacin V®), containing oestrogens (Ovestin®, Blissel®, Colpotrophine®), and reference formulations (Replens®, Universal Placebo). Two in vitro cytotoxicity tests were performed: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red uptake (NRU) upon uterine (HEC-1A), cervical (HeLa) and vaginal (VK2 E6/E7) cells, according to ISO/EN 10993-5 (in vitro evaluation of medical devices). Likewise, a strategy to determine tissue viability on ex vivo porcine vaginal model (through MTT reduction assay and histological analysis) was developed and optimized. The vaginal cell line VK2 E6/E7 conducted to the most accurate calculation of half-maximal Toxic Concentration among all cellson the MTT assay. However it was shown not be sensitive to the NRU assay. Tissues from the porcine model were collected with approx. 15% variability in thickness and variation coefficients lower than 25% when testing negative and positive controls were achieved. These models can improve cost-efficiency in early steps of product development.
  • Effects of buffer composition on site specific glycation of lysine
           residues in monoclonal antibodies
    • Abstract: Publication date: Available online 29 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Alex W. Jacobitz, Andrew B. Dykstra, Chris Spahr, Neeraj J. Agrawal Candidate antibodies under consideration for development as pharmaceuticals must be screened for potential liabilities. Glycation of lysine side chains is one liability which can significantly alter the efficacy of a therapeutic antibody. Antibody candidates are often subjected to stress-testing after purification to assess liabilities that may arise from variability in the manufacturing process to gauge the manufacturability of the molecule. Since previous publications have shown significant site-specific effects of certain buffer components on the glycation rate of individual lysines, we sought to understand the effects of common buffering agents to find suitable buffers for glycation stress-testing (forced glycation). Therapeutic antibodies are typically only exposed to reducing sugars in cell culture media during production, so we sought to identify buffers that could be used as surrogates for media in forced glycation reactions. Our results indicate that common buffering agents can drastically alter the rate of glycation for specific lysines in an antibody. Forced glycation reactions performed in HEPES and citrate buffers both produce site-specific lysine glycation rates that correlate well with cell culture media, whereas bicarbonate buffer has a highly stimulatory effect on most lysines leading to higher total glycation levels and a poor correlation to glycation rates in media.
  • X-ray Diffraction and Theoretical Calculation-Supported Formation of
           Polymorphic Cocrystals Discovered through Thermal Methods: A Case Study
    • Abstract: Publication date: Available online 28 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Zhengzheng Zhou, Mónica Calatayud, Julia Contreras-García, Liang Li, Henry H.Y. Tong, Ying Zheng Polymorphism commonly exists in the preparation of cocrystals and has attracted widespread attention from both the pharmaceutical industry and academia. However, few studies have examined how to discover polymorphic cocrystals and their potential formation mechanism. In this study, we report the novel discovery of salicylic acid: 3-nitrobenzamide (SA-3NBZ) polymorphic cocrystals by thermal methods. The formation mechanism is elucidated based on theoretical calculations. SA-3NBZ polymorphic cocrystals with molar ratio of 1:1 and 2:2 were discovered using the combination of differential scanning calorimetry (DSC) and hot stage microscopy (HSM). Single crystal X-ray diffraction analysis confirmed this discovery. Density functional theory (DFT) calculations corrected with dispersion were conducted to illustrate the energetic stabilization of SA polymorphic cocrystals. Compared with the starting materials, formation of the cocrystals at 1:1 and 2:2 present a weak stabilization with overall energy reduction of -0.01 and -0.05 eV/molecule, respectively. The calculated non-covalent interactions index (NCI) further suggests that intralayer hydrogen bonds and van der Waals forces contribute to these weak interactions. The DFT calculations are in good agreement with the X-ray diffraction data. Hence, thermal analysis is a simple and reliable method to discover polymorphic cocrystals.
  • Cyclization of N-Terminal Glutamic Acid to pyro-Glutamic Acid Impacts
           Monoclonal Antibody Charge Heterogeneity despite Its Appearance as a
           Neutral Transformation
    • Abstract: Publication date: Available online 27 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Zhihua Liu, Joseph Valente, Shujie Lin, Naresh Chennamsetty, Difei Qiu, Mark Bolgar Pyroglutamic acid (pyroGlu) is commonly observed at the N-terminus of therapeutic monoclonal antibodies. Notably, the term “pyroGlu” refers to a single product that could originate from the cyclization of either an N-terminal glutamine or an N-terminal glutamic acid. This is an important and easily overlooked distinction that has major implications on the charge variant nature of a pyroGlu relative to its un-cyclized form. Cyclization of an N-terminal glutamine for instance clearly produces an acidic variant with a lower isoelectric point due to the loss of the positively charged N-terminal amine. In this report, we demonstrate that cyclization of an N-terminal glutamic acid on the other hand produces a basic variant with a higher isoelectric point contrary to the typical assumption that the simultaneous loss of the N-terminal amine and the carboxylic acid side chain would negate the formation of a charge variant. The results of our investigation demonstrate the need to consider the relative strengths of the acidic and basic functional groups which are altered when assessing whether the product will be a charge variant. This study also adds new knowledge and experimental evidence to understand charge heterogeneity in monoclonal antibodies.
  • Development of Coprocessed Chitin-Calcium Carbonate as Multifunctional
           Tablet Excipient for Direct Compression, Part 2: Tableting Properties
    • Abstract: Publication date: Available online 27 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Mohammad Chaheen, Bernard Bataille, Ahmad Yassine, Emmanuel Belamie, Tahmer Sharkawi The use of multifunctional excipients is gaining interest as it simplifies formulations by replacing the need of multiple monofunctional excipients. In previous work, coprocessed Chitin-Calcium carbonate (CC) showed to have good potential as a multifunctional excipient for fast disintegrating tablets produced by direct compression. It allowed for good tablet strength, enhanced powder flowability, higher true and bulk densities with fast disintegrating properties. The objective of this work is to gain insight on CC tableting properties under different tablet manufacturing conditions (different lubrication levels, compression speeds and dwell times) and in formulations with drug models: ibuprofen and paracetamol. Results showed that CC exhibited good tabletability, compressibility and compactibility profiles. CC does not require the addition of lubricant and can be used at high compression speeds and different dwell times. When included in formulations with ibuprofen and paracetamol at different percentages, CC enhanced tablets strength and promoted fast disintegration and drug dissolution. In conclusion, this study shows that CC can be used as a multifunctional excipient (filler-disintegrant-binder) for fast disintegrating tablets produced by direct compression.
  • Forced Oxidative Degradation Pathways of the Imidazole Moiety of
    • Abstract: Publication date: Available online 27 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Yande Huang, Bao-Ning Su, Jonathan Marshall, Scott A. Miller Daclatasvir hydrochloride (DCV) is the active pharmaceutical ingredient of Daklinza, a marketed product for the treatment of hepatitis C viral infection. The intrinsic stability of daclatasvir was evaluated via a forced degradation study. DCV was found to be stable in the solid state. In solution its carbamate moiety is susceptible to basic hydrolysis, while its imidazole is liable to base mediated autoxidation to form degradants 1 and 3, 7-8, respectively. The imidazole moiety can also be oxidized to form degradants 6-7 in the presence of hydrogen peroxide or azobisisobutyronitrile (AIBN). The chloro-adduct degradant 9 was also observed in hydrogen peroxide solution. Furthermore, the imidazole moiety is sensitive to photodegradation in solution. Degradants 2-8 were observed in a solution of DCV exposed to high intensity light/UV light; the formation of degradants 2 and 5-8 were postulated through four degradation pathways. The degradants 3 and 4 were deemed to be secondary degradants of 7 and 5, respectively.
  • Bile duct obstruction leads to increased intestinal expression of breast
           cancer resistance protein with reduced gastrointestinal absorption of
    • Abstract: Publication date: Available online 25 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Takumi Kawanishi, Hiroshi Arakawa, Yusuke Masuo, Noritaka Nakamichi, Yukio Kato Liver dysfunction reduces systemic clearance of drugs that are primarily eliminated by the liver. However, liver dysfunction can cause a reduction in the plasma concentration profiles of certain drugs, including several tyrosine kinase inhibitors (TKIs), after oral administration. The aim of the present study was to clarify the reduction in oral absorption of a TKI, imatinib, and the mechanisms of action involved under conditions of hepatic dysfunction, focusing on intestinal transporters. The maximum plasma concentration (Cmax) of imatinib after oral administration in mice subjected to bile duct ligation (BDL) was lower than that in sham-operated mice, whereas the plasma concentration profile after intravenous administration was essentially unaffected by BDL. The change in Cmax was compatible with a reduction in small intestinal permeability of imatinib observed in the in situ closed loop. Gene expression of abcg2 was increased in BDL mice compared to that in sham-operated mice. Expression of breast cancer resistance protein and P-glycoprotein in the small intestinal brush border membrane fraction from BDL mice was also increased compared to that in sham-operated mice. In summary, the intestinal absorption and permeability of imatinib was decreased in BDL mice, and this may be attributed to the up-regulation of the efflux transporter(s).
  • Improved intestinal mucus permeation of vancomycin via incorporation into
           nanocarrier containing papain-palmitate
    • Abstract: Publication date: Available online 25 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Nuri Ari Efiana, Aida Dizdarević, Christian W. Huck, Andreas Bernkop-Schnürch The aim of this study was to improve intestinal mucus permeation of a peptide antibiotic via incorporation into papain-palmitate modified self-emulsifying drug delivery systems (SEDDS) as nanocarrier. Vancomycin as a peptide antibiotic was lipidized by hydrophobic ion pair (HIP) formation using sodium bis-2-ethylhexyl-sulphosuccinate (SBS) prior to incorporation in SEDDS comprising Capmul MCM, propylenglycol and Kolliphor EL (2:1:2). As mucolytic agent, 0.5% papain-palmitate was introduced in SEDDS formulation containing the vancomycin-SBS ion pair. The formulation was evaluated regarding droplet size, zeta potential and cytotoxicity using Caco-2 cells previous to intestinal mucus permeation studies using Transwell diffusion and rotating tube method. The HIP product yielded from surfactant to drug ratio of 3:1 provided a 25-fold increase in lipophilicity, drug payload in SEDDS of 5% and log DSEDDS/release medium of 2.2. The formulation exhibited a droplet size and zeta potential of 221.5±14.8 nm and -4.2±0.8 mV, respectively. Cytotoxicity study showed that SEDDS formulations were not toxic. Introducing 0.5% papain-palmitate increased the mucus permeability of SEDDS 2.8-fold and 3.3-fold in Transwell diffusion and rotating tube studies, respectively. According to these results, papain decorated SEDDS might be a potential strategy to improve the mucus permeating properties of peptide antibiotics.
  • Characterization of Protein Aggregates, Silicone Oil Droplets, and
           Protein-Silicone Interactions using Imaging Flow Cytometry
    • Abstract: Publication date: Available online 25 May 2019Source: Journal of Pharmaceutical SciencesAuthor(s): Christine Probst Protein therapeutic exposure to siliconized surfaces during manufacturing and storage has potential to induce protein aggregation or generate protein-silicone complexes that are potentially immunogenic. Consequently, assessing the stability and safety of protein therapeutics requires discrimination of protein and silicone oil particles and evaluation of protein-silicone oil interactions. Industry established methods are challenged to distinguish protein aggregates from silicone oil droplets for particles smaller than 5 μm. In addition, emerging techniques for accessing particles in the sub-5 μm range are limited by low sampling volumes, narrow size ranges, complications such as clogging, and an inability to evaluate protein-silicone interactions. In this study, imaging flow cytometry was evaluated as a new method for discrimination of protein aggregates and silicone oil droplets, as well as for quantification of protein-silicone interactions. A simple and fast fluorescence labeling protocol using low concentrations of extrinsic dyes was developed. The results demonstrate that imaging flow cytometry can be used to discriminate fluorescently labeled protein aggregates and silicone oil droplets with greater than 95% accuracy, regardless of size, and protein-silicone oil interactions can be assessed qualitatively through inspection of image data or quantitatively using features extracted from the image data.
  • Physiologically Based Pharmacokinetic Modeling to Supplement Nilotinib
           Pharmacokinetics and Confirm Dose Selection in Pediatric Patients
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Tycho Heimbach, Wen Lin, Florence Hourcade-Potelleret, Xianbin Tian, Francois Pierre Combes, Nicholas Horvath, Handan He In adult patients, nilotinib is indicated for chronic myeloid leukemia at an approved oral dose of 300 or 400 mg BID. Physiologically based pharmacokinetic (PBPK) model was developed to describe and supplement limited PK data in the pediatric population ranging from 2 to less than 6 years of age and ultimately inform dosing regimen. An adult Simcyp PBPK model was established and verified with clinical pharmacokinetic data after a single or multiple oral doses of 400 mg nilotinib (230 mg/m2). The model was then applied to a pediatric PBPK model, taking account of ontogeny profiles of metabolizing enzymes and pediatric physiological parameters. The model was further verified using observed pediatric PK data in 12- to
  • Influx and Efflux Transporters Contribute to the Increased Dermal Exposure
           to Active Metabolite of Regorafenib After Repeated Oral Administration in
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Aya Hasan Al-Shammari, Yusuke Masuo, Ken-ichi Fujita, Yuka Yoshikawa, Noritaka Nakamichi, Yutaro Kubota, Yasutsuna Sasaki, Yukio Kato The multikinase inhibitor regorafenib, which is a standard treatment for certain cancer patients after disease progression following other approved therapies, exhibits delayed-onset dermal toxicity. Here, we aimed to clarify the mechanisms that contribute to the increased dermal exposure to active metabolite M-5 of regorafenib after repeated oral administration. The dermal concentration of M-5 at 24 h after the last 5 oral administrations of regorafenib in mdr1a/1b/bcrp-/- mice was more than 190 times that in wild-type mice. The skin-to-plasma concentration ratio of M-5 in mdr1a/1b/bcrp-/- was also higher than in wild-type mice, suggesting possible involvement of P-glycoprotein and breast cancer resistance protein in regulating the dermal distribution. The area under the plasma concentration-time curve values of M-5 and its precursor M-2 in plasma of mdr1a/1b/bcrp-/- were at most 26 and 3 times those in wild-type mice, respectively. Interestingly, repeated administration of regorafenib markedly increased the area under the plasma concentration-time curve of M-5 in plasma, but not liver, compared with a single dose. Intravenous administration of M-5 dose-dependently reduced the liver-to-plasma concentration ratio. Our results indicate that hepatic uptake of M-5 may partially explain the accumulation of M-5 in the systemic circulation, but multiple factors, including influx and efflux transporters, are involved in determining dermal exposure to M-5.
  • Hyperhydration Effect on Pharmacokinetic Parameters and Detection
           Sensitivity of Recombinant Human Erythropoietin in Urine and Serum Doping
           Control Analysis of Males
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Ioanna Athanasiadou, Aristeidis Dokoumetzidis, Sven Christian Voss, Wesal El Saftawy, Mohammed Al-Maadheed, Georgia Valsami, Costas Georgakopoulos Excessive fluid intake, that is, hyperhydration, may be adopted by athletes as a masking method during antidoping sample collection to influence the excretion patterns of doping agents and, therefore, manipulate their detection. The aim of this exploratory study was to assess the hyperhydration effect on the detection sensitivity of recombinant human erythropoietin (rHuEPO) by sodium N-lauroyl sarcosinate (“sarkosyl”) polyacrylamide gel electrophoresis analysis. The influence of hyperhydration on the serum and urinary pharmacokinetic (PK) profiles of rHuEPO was also investigated. Seven healthy physically active nonsmoking Caucasian males participated in a 31-day clinical study comprising a baseline (days 0, 1-3, and 8-10) and a drug phase (days 15-17, 22-24, and 29-31). Epoetin beta was administered subcutaneously at a single dose of 3000 IU on days 15, 22, and 29. Hyperhydration was applied in the morning on 3 consecutive days (days 1-3, 8-10, 22-24, and 29-31), that is, 0, 24, and 48 h after first fluid ingestion. Water and a commercial sports drink were used as hyperhydration agents (20 mL/kg body weight). Serum and urinary concentration-time profiles were best described by a one-compartment PK model with zero-order absorption. Delayed absorption was observed after hyperhydration and, therefore, lag time was introduced in the PK model. Results showed no significant difference (p> 0.05) on serum or urinary erythropoietin concentrations under hyperhydration conditions. A trend for decreasing volume of distribution and increasing clearance after hyperhydration was observed, mainly after sports drink consumption. However, no significant differences (p> 0.05) due to hyperhydration for any of the serum PK parameters calculated by noncompartmental PK analysis were observed. Renal excretion of endogenous erythropoietin and rHuEPO, as reflected on the urinary cumulative amount, was increased approximately twice after hyperhydration and this supports the nonsignificant difference on the urinary concentrations. Analysis of serum and urine samples was able to detect rHuEPO up to 72 h after drug administration. The detection window of rHuEPO remained unaffected after water or sports drink ingestion. Hyperhydration had no effect on the detection sensitivity of EPO either in serum or urine samples.
  • Physical Characterization of Halofantrine-Encapsulated Fat Nanoemulsions
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Iman Haidar, Ian H. Harding, Ian C. Bowater, Alasdair W. McDowall We report the colloidal characterization of halofantrine (Hf)-laden soybean oil fat emulsions. Hf increased the zeta potential, at all pH values, of the fat emulsions. Concomitant with this, the isoelectric point (i.e.p.) of the emulsion increased to higher pH values. The emulsion was destabilized by a small amount of Hf; interestingly, however, this was ameliorated by increasing the amount of Hf. The particle size and polydispersity of the fat emulsion reflected this with a small Hf concentration resulting in a significant increase in both particle size and polydispersity, but less so as the Hf concentration was increased. Emulsions lost stability as the pH approached the i.e.p. and this effect was greatest for the small Hf concentration emulsions. Cryogenic transmission electron microscopy showed the presence of beading or string-like behavior leading to gross distortions of the spherical shape for highly unstable emulsions. We conclude that to maintain good stability for Hf-laden soybean oil emulsions, the pH of the emulsion should be kept away from its i.e.p, and also that the drug concentration should be maintained at a relatively high value.
  • A Delivery System for Oral Administration of Proteins/Peptides Through
           Bile Acid Transport Channels
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Siwen Wu, Wen Bin, Biyun Tu, Xifeng Li, Wei Wang, Suling Liao, Changshan Sun Proteins and peptides are poorly absorbed via oral administration because of the gastrointestinal tract environment and lysosomal digestion after apical endocytosis. A delivery system, consisting of a deoxycholic acid–conjugated nanometer-sized carrier, may enhance the absorption of proteins in the intestine via the bile acid pathway. Deoxycholic acid is first conjugated to chitosan. Liposomes are then prepared and loaded with the model drug insulin. Finally, the conjugates are bound to the liposome surface to form deoxycholic acid and chitosan conjugate–modified liposomes (DC-LIPs). This study demonstrates that DC-LIPs can promote the intestinal absorption of insulin via the apical sodium-dependent bile acid transporter, based on observing fluorescently stained tissue slices of the rat small intestine and a Caco-2 cell uptake experiment. Images of intestinal slices revealed that excellent absorption of DC-LIPs is achieved via apical sodium-dependent bile acid transporter, and a flow cytometry experiment proved that DC-LIPs are a highly efficient delivery carrier. Caco-2 cells were also used to study the lysosome escape ability of DC-LIPs. We learned from confocal microscopy photographs that DC-LIPs can protect their contents from being destroyed by the lysosome. Finally, according to pharmacokinetic analyses, insulin-loaded DC-LIPs show a significant hypoglycemic effect with an oral bioavailability of 16.1% in rats with type I diabetes.
  • Liquid Oil Marbles: Increasing the Bioavailability of Poorly Water-Soluble
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Petra Janská, Ondřej Rychecký, Aleš Zadražil, František Štěpánek, Jitka Čejková Many new therapeutic candidates and active pharmaceutical ingredients (APIs) are poorly soluble in an aqueous environment, resulting in their reduced bioavailability. A promising way of enhancing the release of an API and, thus, its bioavailability seems to be the use of liquid oil marbles (LOMs). An LOM system behaves as a solid form but consists of an oil droplet in which an already dissolved API is encapsulated by a powder. This study aims to optimize the oil/powder combination for the development of such systems. LOMs were successfully prepared for 15 oil/powder combinations, and the following properties were investigated: particle mass fraction, dissolution time, and mechanical stability. Furthermore, the release of API from both LOMs and LOMs encapsulated into gelatine capsules was studied.
  • Overcoming Poor Tabletability of Bulky Absorption Enhancers by Spray
           Drying Technology
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Weiwei Fan, Aohua Wang, Yue Wu, Jorrit J. Water, Stephen T. Buckley, Lars Hovgaard, Mingshi Yang, Yong Gan Absorption enhancers are often a major component of solid oral peptide formulations as compared to the active pharmaceutical ingredient and excipients. This commonly results in poor tabletability that is hard to mitigate in direct compaction by addition of small amounts of excipients. To improve the tabletability of bulky absorption enhancers, the model absorption enhancers, sodium cholate and deoxycholic acid, were co–spray-dried with hydroxypropyl methylcellulose E5, where the percentage of absorption enhancers was not lower than 90% (w/w). The physicochemical properties of the resulting powders were assessed by laser diffraction, scanning electron microscopy, X-ray powder diffraction, thermogravimetric analysis, and differential scanning calorimetry. The powders were compressed into tablets, and the tabletability was evaluated. Co–spray drying with 10% of hydroxypropyl methylcellulose significantly improved the tabletability of the both absorption enhancers. Moreover, it was demonstrated that small particle size and amorphous state rather than high moisture content contributed to the improved tabletability of the spray-dried powders. The study suggests that spray drying technology can be promising to overcome the poor tabletability of oral peptide formulation consisting of large amounts of absorption enhancers.
  • Prediction of Dissolution Profiles From Process Parameters, Formulation,
           and Spectroscopic Measurements
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Yuxiang Zhao, Wenlong Li, Zhenqi Shi, James K. Drennen, Carl A. Anderson This study utilized multiple modeling approaches to predict immediate release tablet dissolution profiles of 2 model drugs: theophylline and carbamazepine. Two sets of designs of experiments were applied based on individual drug characteristics to build in adequate dissolution variability. The tablets were scanned using a near-infrared (NIR) spectrometer and then subjected to in vitro dissolution test at critical time points. Because of the inherent difference in dissolution profiles, a hierarchical modeling approach was applied for theophylline data, whereas global models were constructed from carbamazepine data. The partial least squares models were trained using 3 predictor sets including (1) formulation, material, and process variables, (2) NIR spectra, and (3) a combination of both. The dependent variables of the models were the dissolution profiles, which were presented either as parameters of Weibull fitting curves or raw data. The comparison among the predictive models revealed that the incorporation of NIR spectral information in calibration reduced prediction error in the carbamazepine case but undermined the performance of theophylline models. It suggests that the modeling strategy for dissolution prediction of pharmaceutical tablets should not be universal but on a case-by-case basis.
  • Development and Validation of a Discriminatory Dissolution Method for
           Rifaximin Products
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Sathish Dharani, Sogra F. Barakh Ali, Hamideh Afrooz, Mansoor A. Khan, Ziyaur Rahman The commercial product of rifaximin (RFX) contains α form. The α form can change to β form on exposure to high humidity that can occur during manufacturing, stability, and in-use period. It is critical to maintain α form of the drug in a drug product to avoid variability in clinical response. U.S. Food and Drug Administration dissolution method was found to be nondiscriminatory for RFX formulations containing either 100% α or β form. The objective of this study was to develop a discriminatory dissolution method that can detect low levels of α to β transformation in RFX products. Formulations containing a variable fraction of α and β forms were prepared by using direct compression method. Dissolution parameters investigated were type of dissolution medium (water and phosphate buffer), volume (500, 900, and 1000 mL), and paddle speed (50, 75, and 150 rpm). Dissolution in water with 0.2% sodium lauryl sulfate was less than 80% and nondiscriminatory. However, dissolution tested in a phosphate buffer pH 7.4 with 0.2% sodium lauryl sulfate at 50 rpm was discriminatory with more than 17.5% difference in dissolution profile between formulations containing α and β forms. The developed method can detect polymorphic transformation if there is 25% or more β form conversion.
  • Folate-Targeted Polyethylene Glycol–Modified Photosensitizers
           for Photodynamic Therapy
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Qian Liu, Jin Wang, Shuang Li, Guojie Li, Qingle Chen, Zhangyong Hong Pyropheophorbide a (Pyro) is a promising photosensitizer; however, it has no tumor selectivity and enrichment capability. In our former work, the prepared folic acid (FA)-Pyro conjugates showed considerably improved tumor accumulation and photodynamic therapy (PDT) activity in cell- and animal-based studies. However, the targeting capability, selectivity and water solubility of the conjugate remain problematic. Here, we evaluated the installation of hydrophilic polyethylene glycol chains as the linker between Pyro and FA, by avoiding direct conjugation of Pyro with FA, aiming to improve tumor selectivity and accumulation. However, PEGylation may have negative effects on the PDT activity and cutaneous phototoxicity. Therefore, we chose various lengths of PEGs as linkers to optimize the molecular weight, hydrophilicity, and PDT activity and, thus, to balance the tumor selectivity and biological function of the conjugate. One optimized conjugate, Pyro-PEG1K-FA, exhibited excellent tumor enrichment and was able to eradicate subcutaneous tumors at a considerably reduced dose. We report the synthesis and characterization of these conjugates as well as the evaluation of their tumor accumulation ability and the corresponding PDT efficiency through in vitro and in vivo experiments.
  • Modeling of Semicontinuous Fluid Bed Drying of Pharmaceutical Granules
           With Respect to Granule Size
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Michael Ghijs, Elisabeth Schäfer, Ashish Kumar, Philippe Cappuyns, Ivo Van Assche, Fien De Leersnyder, Vaĺerie Vanhoorne, Thomas De Beer, Ingmar Nopens In the transition of the pharmaceutical industry from batchwise to continuous drug product manufacturing, the drying process has proven challenging to control and understand. In a semicontinuous fluid bed dryer, part of the ConsiGma™ wet granulation line, the aforementioned production methods converge. Previous research has shown that the evolution of moisture content of the material in this system shows strong variation in function of the granule size, making the accurate prediction of this pharmaceutical critical quality attribute a complex case. In this work, the evolution of moisture content of the material in the system is modeled by a bottom-up approach. A single granule drying kinetics model is used to predict the moisture content evolution of a batch of material of a heterogeneous particle size, where it is the first time that the single granule drying mechanism is validated for different granule sizes. The batch approach was validated when the continuous material inflow rate and filling time of the dryer cell are constant. The original single granule drying kinetics model has been extended to capture the granules’ equilibrium moisture content. Finally, the influence of drying air temperature is captured well with a droplet energy balance for the granules.
  • Mechanism of Drug Release From Temperature-Sensitive Formulations Composed
           of Low-Melting-Point Microcrystalline Wax
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Kohei Matsumoto, Shin-ichiro Kimura, Shuji Noguchi, Shigeru Itai, Hiromu Kondo, Yasunori Iwao It was reported that wax matrix (WM) particles composed of low-melting-point microcrystalline wax showed unique release behaviors; the particles released only a small amount of the entrapped drug (non–diffusion-controlled release) at 37°C, whereas it showed comparatively fast drug release in a diffusion-controlled manner at 25°C. However, the mechanism of the drug release is still unclear. The objective of this study was to determine the mechanism of drug release from the WM particles using X-ray computed tomography. In the WM particles collected during dissolution tests at 25°C, the void space derived from drug release increased with increasing time, and there was no change in the structure, indicating that the WM particles released drug while maintaining the particle shape at 25°C. In the WM particles collected during dissolution tests at 37°C, the void space was confirmed at initial time point; however, at subsequent time points, the void space was disappeared, and the roughness of the surface was evident. This structural change may have blocked the conveyance pathway of the outer medium, which would inhibit the drug release. The difference between the drug-release mechanisms of the WM particles at the 2 temperatures will be valuable for developing cooling-triggered, temperature-sensitive formulations.
  • Bulk Dynamic Spray Freeze-Drying Part 2: Model-Based Parametric Study for
           Spray-Freezing Process Characterization
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Israel B. Sebastião, Bakul Bhatnagar, Serguei Tchessalov, Satoshi Ohtake, Matthias Plitzko, Bernhard Luy, Alina Alexeenko Spray freeze-drying is an evolving technology that combines the benefits of spray-drying and conventional lyophilization techniques to produce drug substance and drug product as free-flowing powders. The high surface-to-volume ratio associated to the submillimeter spray-frozen particles contributes to shorter drying and reconstitution times. The formation of frozen particles is the most critical part of this dehydration technique because it defines the properties of final product. Based on a previously proposed and validated model, the current goal is to understand the role of various controllable parameters in the spray-freezing process. More specifically, given a set of spraying conditions, the model is used to predict the minimum distance required to cool and freeze the droplets below a temperature that prevents coalescence and product agglomeration. A parametric study is carried out to map the operational limit conditions of the actual spray-freezing column apparatus under consideration. For the spray freeze-drying conditions of interest, model simulations indicate that convection contributes to at least 80% of the total droplet heat transfer and, consequently, that freezing column gas temperature and droplet diameter are the most important process parameters affecting the freezing distance.
  • Bulk Dynamic Spray Freeze-Drying Part 1: Modeling of Droplet Cooling and
           Phase Change
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Israel B. Sebastião, Bakul Bhatnagar, Serguei Tchessalov, Satoshi Ohtake, Matthias Plitzko, Bernhard Luy, Alina Alexeenko In spray freeze-drying (SFD), the solution is typically dispersed into a gaseous cold environment producing frozen microparticles that are subsequently dried via sublimation. This technology can potentially manufacture bulk lyophilized drugs at higher rates compared with conventional freeze-drying in trays and vials because small frozen particles provide larger surface area available for sublimation. Although drying in SFD still has to meet the material collapse temperature requirements, the final characteristics of the respective products are mainly controlled by the spray-freezing dynamics. In this context, the main goal of this work is to present a single droplet spray-freezing model and validate it with previously published simulations and experimental data. For the investigated conditions, the droplet temperature evolutions predicted by the model agree with experiments within an error of ±10%. The proposed engineering-level modeling framework is intended to assist future development of efficient SFD processes and support scale up from laboratory to commercial scale equipment.
  • Physical Stability of an Amorphous Sugar Matrix Dried From Methanol as an
           Amorphous Solid Dispersion Carrier and the Influence of Heat Treatment
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Koji Takeda, Takanari Sekitoh, Akiho Fujioka, Kayoko Yamamoto, Takashi Okamoto, Tsutashi Matsuura, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura An amorphous sugar matrix, after drying from an organic solvent, was investigated for use as a method for dispersing hydrophobic drugs (solid dispersion). However, the amorphous sugar, originally contained in the organic solvent, had a significantly low glass transition temperature (Tg), thus rendering it physically unstable. In this study, we examined the physicochemical properties of a sugar in a dried matrix and in an organic solvent, using α-maltose and methanol as a representative sugar and organic solvent. The apparent molar volume of α-maltose was ∼30% smaller in methanol than in water. The methanol-originated amorphous α-maltose exhibited a much greater degree of hydrogen bonding than the water-originated one. Considering these findings, we conclude that the α-maltose maintained its compact conformation in the dried state and consequently caused the markedly low Tg. Second, it was found that heating under appropriate conditions resulted in an increase in the Tg of the methanol-originated amorphous α-maltose as well as a decrease in the level of hydrogen bonding. The aqueous dissolution of 2 model hydrophobic drugs (indomethacin and ibuprofen) from the solid dispersion was also improved as the result of the heat treatment, whereas, to the contrary, the dissolution of another model drug (curcumin) was lowered.
  • Orthogonal Redundant Monitoring of a New Continuous Fluid-Bed Dryer for
           Pharmaceutical Processing by Means of Mass and Energy Balance Calculations
           and Spectroscopic Techniques
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Victoria Pauli, Frantz Elbaz, Peter Kleinebudde, Markus Krumme In line with the ongoing shift from batch to continuous pharmaceutical production of solid oral dosage forms, a novel continuous fluid-bed dryer was developed. The forced feed nature of the Glatt GPCG2 CM fluid-bed dryer allows continuous, first-in-first-out drying of wet granulate materials based on its compartmentalized, rotating fluidizing chamber. The presented work aims to introduce the dryer’s functionalities in detail, and to demonstrate that the rotating fluid-bed chambers facilitates a stable drying behavior, which ensures robust and repeatable residual moisture contents (loss-on-drying [LOD]) of the discharged granules. Furthermore, a mass and energy balance (MEB) is derived, based on the logged process values of the granulating and drying units. Two independent test experiments demonstrate that precise LOD prediction in real time is achievable by MEB to serve as an orthogonal process analytical technology method to common near-infrared spectroscopy. On average, MEB results differed by 0.36% LOD (absolute) from offline reference analyses, and by 0.61% LOD from predictions made with an in-house available near-infrared spectroscopy method. Furthermore, good correlation between the observed and expected thermal energy loss was found. The derived MEB is solely based on physical principles; hence it is product independent and transferable to other materials that are processed on the described equipment.
  • Solubility and Permeability Improvement of Quercetin by an Interaction
           Between α-Glucosyl Stevia Nanoaggregates and Hydrophilic Polymer
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Hiromasa Uchiyama, Yuhei Wada, Yuta Hatanaka, Yoshiyuki Hirata, Masahiko Taniguchi, Kazunori Kadota, Yuichi Tozuka The effect of composite formation between α-glucosyl stevia (Stevia-G) and hydrophilic polymers on solubility and permeability enhancement of quercetin hydrate (QUE) was evaluated. Polyvinylpyrrolidone K-30 (PVP), hydroxypropyl methylcellulose 2910-E (HPMC), and hydroxypropyl cellulose SSL (HPC) were selected as candidate hydrophilic polymers. Fluorescence studies with pyrene and curcumin suggested composite formation occurs between Stevia-G aggregate and polymers. Furthermore, the strength of interaction between Stevia-G aggregate and polymers was as follows: PVP> HPMC> HPC. Evaporated particles (EVPs) of QUE with Stevia-G and polymers showed synergic QUE solubility enhancement. Solubility of QUE from the EVPs was enhanced in the following order: Stevia-G/PVP> Stevia-G/HPMC> Stevia-G/HPC, in accordance with the degree of interaction. Enhanced membrane permeability of QUE from the EVPs of Stevia-G/PVP was confirmed using Caco-2 cells. The amount of QUE that permeated Caco-2 cells from the EVPs of Stevia-G/PVP was 13.7-, 4.7-, and 2.1-fold higher than that of the untreated QUE powder, EVPs of Stevia-G, and EVPs of PVP, respectively. These results indicated that the composite formed by Stevia-G and PVP can dramatically enhance the solubility and membrane permeability of QUE.
  • Drug Carrier-Oriented Polygeline for Preparing Novel Polygeline-Bound
           Paclitaxel Nanoparticles
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Kaibin Xiong, Jianyang Wu, Yang Liu, Na Wu, Jinlan Ruan Polygeline is a highly promising drug carrier-oriented material for important applications in pharmacy field due to its low-cost and unique properties similar to albumin. In this study, polygeline-bound paclitaxel nanoparticles (Npb-PTXS) were fabricated through a combination of low-pressure emulsification and high-pressure homogenization. The effects of a series of production parameters on mean particle size, particle size distribution and drug loading of Npb-PTXS were systematically evaluated. The characteristics of Npb-PTXS, such as surface morphology, physical status of paclitaxel (PTX) in Npb-PTXS, redispersibility of Npb-PTXS in purified water and bioavailability in vivo were also investigated. It is revealed that the optimal preparation conditions included an aqueous phase pH value of about 6.5, protein mass concentration of 0.33%, with mass ratio of PTX to protein of 30%, high pressure of 1200 bar, high-pressure passes of 25 times and low-pressure emulsifying passes of 20 times. Obtained Npb-PTXS shows good resolubility compared to commercially available Abraxane®, containing round or oval shaped particles with mean particle size of around 188.3 nm, polydispersity index of 0.163 and zeta potential of −31.1 mV. PTX in Npb-PTX is amorphous, and its content is approximately 12.04%. Encapsulation efficiency of Npb-PTXS reaches 81.2%. Moreover, in vivo pharmacokinetic studies showed that the intravenous relative bioavailability of Npb-PTXS to Abraxane was 83.89%.
  • A Kinetic Approach to Determining Drug Distribution in Complex Biphasic
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Yixuan Dong, Leanna Hengst, Deval Patel, Robert Hunt, Haiou Qu, Stephanie Choi, Muhammad Ashraf, Celia N. Cruz, Xiaoming Xu Pharmaceutical emulsions contain multiple components, such as micellar, aqueous, and oil phases, leading to complex drug transfer and equilibrium phenomena. These complex components present challenges for the bioequivalence assessment of the drug products. The objective of the study was to develop a method that can probe the underlying mechanism and process of drug distribution. The concept of drug partitioning into biphasic systems was used to simplify the complex transfer phenomenon. A kinetic method was developed taking into account the biphasic diffusion. Using this approach, both the rate (kinetics) and the extent (equilibrium) of distribution can be determined. For method development purpose, 3 model compounds (triamcinolone acetonide, difluprednate, and cyclosporine), with expected partition coefficient values ranging from 2 to 6, were tested using the kinetic method and the traditional shake-flask method. The values obtained by the 2 methods for all compounds correlated well (r2 = 0.825). Various organic and aqueous solvents which are commonly encountered in formulations were also tested to determine the impact of phase composition on drug distribution. The kinetic method was found to offer more flexibility in terms of solvent composition and can lead to better understanding for drug distribution and potential drug release in complex biphasic systems.
  • Modeling the Air Pressure Increase Within a Powder Bed During
           Compression—A Step Toward Understanding Tablet Defects
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Gerard R. Klinzing, Gregory M. Troup The cause of tablet defects, such as cracking, bubbling, and capping, during compression is currently not fully understood. Prior experimental work suggests that an increase in internal air pressure on powder compression can directly contribute to the formation of cracks within a tablet. The present study examines the air pressure increase on compression in a fully two-dimensional axisymmetric tablet geometry while being coupled to a plasticity model describing the evolution of tablet relative density on consolidation. It is shown numerically that increasing compression speed results in a large air pressure increase on the order of 1-1.5 MPa which approaches the diametrical tensile strength of tablets. In addition, it is shown experimentally through X-ray microcomputed tomography scans of tablets made at various dwell times that increasing dwell times equivalent to that on a tablet press has no effect on the degree of cracking within the tablet. Only when dwell times reach a time scale of 10 to 100 s does the air pressure diminish to a point at which cracking is eliminated. The reduction in air pressure during these extended dwells is captured by the current model. The experimental and numerical work presented here couples for the first time an air pressure model and plasticity model on compression. In addition, it provides a foundation for understanding how realistic tableting aspects such as precompression and tablet size impact the air pressure increase on consolidation.
  • Incorporation With Dendrimer-Like Biopolymer Leads to Improved Soluble
           Amount and In Vitro Anticancer Efficacy of Paclitaxel
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Ying Xie, Yuan Yao The aim of this study was to explore the solubilization efficacy of octenylsuccinate hydroxypropyl phytoglycogen (OHPP), an amphiphilic dendrimer-like biopolymer, in improving the soluble amount and efficacy of paclitaxel (PTX), a potent anticancer active pharmaceutical ingredient. PTX was incorporated with OHPP in the form of solid dispersion (PTX-OHPP SD), which was characterized for its PTX crystallinity, interactions between PTX and OHPP, PTX soluble amount and dissolution profile, and in vitro inhibitory efficacy against cancer cell lines. The incorporation with OHPP led to the amorphous state of PTX. FTIR spectra showed potential hydrogen bonding between PTX and OHPP, and hydrophobic interaction could play an important role in the association of PTX molecules with OHPP particulates. The soluble amount of PTX with PTX-OHPP SD was>14,000 times that of PTX alone. During dissolution, over 70% PTX dissolved in 5 min and this soluble amount was maintained for at least 180 min. With the 3 cancer cell lines tested, the PTX of PTX-OHPP SD showed a much lower IC50 than the dimethyl sulfoxide–assisted PTX solutions. Based on the result, the mechanism of using OHPP as a highly potent solubilizer for PTX was discussed.
  • Conjugation of Emtansine Onto Trastuzumab Promotes Aggregation of the
           Antibody–Drug Conjugate by Reducing Repulsive Electrostatic Interactions
           and Increasing Hydrophobic Interactions
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Aditya V. Gandhi, Theodore W. Randolph, John F. Carpenter The impact of drug conjugation on intra- and intermolecular interactions of trastuzumab (TmAb) was determined by comparing the conformational and colloidal stabilities of TmAb and trastuzumab emtansine (T-DM1). In low ionic strength formulations, drug conjugation to native lysine residues of TmAb significantly reduced the repulsive electrostatic interactions between T-DM1 molecules. When these electrostatic interactions were screened in solutions with high ionic strength, intermolecular interactions between T-DM1 molecules were found to be more attractive than those between TmAb molecules. Drug conjugation lowered the colloidal stability of T-DM1 compared to TmAb, making T-DM1 more susceptible to agitation-induced aggregation. The presence of polysorbate-20 in the formulations inhibited aggregation of TmAb and T-DM1 induced by the hydrophobic air-water interface. Furthermore, the effect of increased hydrophobic interactions between T-DM1 molecules was studied by monitoring aggregation in TmAb and T-DM1 solutions that were incubated at 4°C, 25°C, and 50°C. Conjugating DM1 to TmAb increased the hydrophobicity of the molecule, and faster aggregation of T-DM1 at 50°C could be attributed to a temperature-dependent increase in hydrophobic interactions between T-DM1 molecules.
  • Rapid Prediction of Deamidation Rates of Proteins to Assess Their
           Long-Term Stability Using Hydrogen Exchange–Mass Spectrometry
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Chamalee L.D. Gamage, Tyler S. Hageman, David D. Weis Deamidation is an important degradation pathway for proteins. Estimating deamidation propensities is essential for predicting their long-term stability. However, predicting deamidation rates in folded proteins is challenging because higher-order structure has a significant and unpredictable effect on deamidation. Here, we investigated the correlation between amide hydrogen exchange (HX) and deamidation to assess the potential of using hydrogen exchange–mass spectrometry (HX-MS) to rapidly predict deamidation propensity. Maltose-binding protein and a structurally less stable mutant, W169G, were stored in the dark at pH 7.0 at 23 ± 2°C for 1 year. Deamidation at each asparagine site was measured using liquid chromatography–mass spectrometry after trypsin digestion. Deamidation rates at each deamidation site were determined based on first-order kinetics. HX rates at the deamidation sites were determined before storage using the shortest peptic peptide containing each site using conventional bottom-up HX-MS at pD 7.0 at 25°C. We observed a power law correlation between deamidation half-life and HX half-life for the NG sites with measurable kinetics. For NA sites, slow deamidation was only observed at 2 sites located in rapidly exchanging regions. Our findings demonstrate that HX-MS can be used to reliably and rapidly rank deamidation propensity in folded proteins.
  • Impact of Precipitation of Antibody Therapeutics After Subcutaneous
           Injection on Pharmacokinetics and Immunogenicity
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Francis Kinderman, Brittany Yerby, Vibha Jawa, Marisa K. Joubert, Nathan H. Joh, Jennifer Malella, Johnathan Herskovitz, Jiansong Xie, John Ferbas, Helen J. McBride Antibody therapeutics with poor solubility in the subcutaneous matrix may carry unintended risks when administered to patients. The objective of this work was to estimate the risk of antibodies that precipitate in vitro at neutral pH by determining the impact of poor solubility on distribution of the drug from the injection site as well as immunogenicity in vivo. Using fluorescence imaging in a mouse model, we show that one such precipitation-prone antibody is retained at the injection site in the subcutaneous space longer than a control antibody. In addition, we demonstrate that retention at the injection site through aggregation is concentration-dependent and leads to macrophage association and germinal center localization. Although there was delayed disposition of the aggregated antibody to draining lymph nodes, no overall impact on the immune response in lymph nodes, systemic exposure of the antibody, or enhancement of the anti-drug antibody response was evident. Unexpectedly, retention of the precipitated antibody in the subcutaneous space delayed the onset of the immune response and led to an immune suppressive response. Thus, we conclude that precipitation due to poor solubility of high doses of antibody formulations delivered subcutaneously may not be of special concern in terms of exposure or immunogenicity.
  • Probing the Tryptophan Environment in Therapeutic Proteins: Implications
           for Higher Order Structure on Tryptophan Oxidation
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Gregory V. Barnett, Gurusamy Balakrishnan, Naresh Chennamsetty, Laurel Hoffman, Jacob Bongers, Li Tao, Yunping Huang, Thomas Slaney, Tapan K. Das, Anthony Leone, Sambit R. Kar Tryptophan (Trp) oxidation in proteins leads to a number of events, including changes in color, higher order structure (HOS), and biological activity. We describe here a number of new findings through a comprehensive characterization of 6 monoclonal antibodies (mAbs) following selective oxidation of Trp residues by 2,2'-azobis(2-amidinopropane) dihydrochloride. Fluorescence spectroscopy, in combination with second derivative analysis, demonstrates that the loss of Trp fluorescence intensity is a sensitive indicator of Trp oxidation in mAbs. Size-exclusion chromatography with UV and intrinsic Trp fluorescence detection was demonstrated to be a useful method to monitor Trp oxidation levels in mAbs. Furthermore, the Trp oxidation levels measured by size-exclusion chromatography with UV and intrinsic Trp fluorescence detection were found to be in agreement with the values obtained from tryptic peptide mapping by liquid chromatography with mass spectrometric detection and correlate with the total solvent accessible surface area of the exposed Trp residues from in silico modeling. Finally, near-UV circular dichroism and Raman spectroscopy were used to evaluate the impact of Trp oxidation on HOS and identify specific oxidation products, respectively. This work demonstrates that protein HOS is altered on Trp oxidation in mAbs and multiple spectroscopic markers can be used to monitor the molecule-dependent Trp oxidation behavior.
  • Cyclodextrin Reduces Intravenous Toxicity of a Model Compound
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Priscilla Mantik, Minli Xie, Harvey Wong, Hank La, Ronald W. Steigerwalt, Uday Devanaboyina, Geoffrey Ganem, Danny Shih, John A. Flygare, Wayne J. Fairbrother, Paroma Chakravarty, David Russell, Gilberto E. Fernandez, Ajit S. Narang Solubilization of new chemical entities for toxicity assessment must use excipients that do not negatively impact drug pharmacokinetics and toxicology. In this study, we investigated the tolerability of a model freebase compound, GDC-0152, solubilized by pH adjustment with succinic acid and complexation with hydroxypropyl-β-cyclodextrin (HP-β-CD) to enable intravenous use. Solubility, critical micelle concentration, and association constant with HP-β-CD were determined. Blood compatibility and potential for hemolysis were assessed in vitro. Local tolerability was assessed after intravenous and subcutaneous injections in rats. A pharmacokinetic study was conducted in rats after intravenous bolus administration.GDC-0152 exhibited pH-dependent solubility that was influenced by self-association. The presence of succinic acid increased solubility in a concentration-dependent manner. HP-β-CD alone also increased solubility, but the extent of solubility enhancement was significantly lower than succinic acid alone. Inclusion of HP-β-CD in the solution of GDC-0152 improved blood compatibility, reduced hemolytic potential by ∼20-fold in vitro, and increased the maximum tolerated dose to 80 mg/kg.
  • The Role of Surface Active Agents in Ophthalmic Drug Delivery:
           A Comprehensive Review
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Shaimaa S. Ibrahim With the significant advances made in nanotechnology, research efforts focused on developing novel drug delivery platforms that can overcome the multitude of challenges encountered in ophthalmic drug delivery. Surface active agents (SAAs) have been extensively used for the formulation of many of the dosage forms targeting ocular tissues. Novel ophthalmic carriers utilizing SAAs were broadly classified into particulate, vesicular, and controlled release drug delivery systems. Depending on their physicochemical properties, SAAs can perform a variety of roles ranging from wetting agents, emulsifiers, stabilizers, charge inducers, solubilizers, antimicrobial agents, corneal permeation enhancers, and gelling agents. Nevertheless, their use is limited by their potential toxicity and possible interactions with other formulation ingredients. This review provides a comprehensive analysis of the different functional roles of SAAs in novel ophthalmic drug delivery platforms, their mechanism of action, and limitations that need to be considered during formulation to maximize their potential benefit. Understanding the mechanisms by which they perform their different roles and the possible interactions between SAAs and other formulation ingredients can help orientate the choice of formulators toward the SAA most suitable for the intended ocular application at a concentration that is both safe and effective.
  • Editorial Advisory Board
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s):
  • Whole-Body Physiologically Based Pharmacokinetic Modeling of Trastuzumab
           and Prediction of Human Pharmacokinetics
    • Abstract: Publication date: June 2019Source: Journal of Pharmaceutical Sciences, Volume 108, Issue 6Author(s): Dong-Jun Bae, Sang-Yeob Kim, Sang Mun Bae, Ae-Kyung Hwang, Kwan Cheol Pak, SeokKyu Yoon, Hyeong-Seok Lim In the present study, we evaluated the pharmacokinetics (PK) of trastuzumab and sought to predict human PK based on animal studies, through the use of optical imaging and a whole-body physiologically based pharmacokinetic (WB-PBPK) modeling approach. The PK study was conducted in 24 mice, where serial blood samples were withdrawn and major organs were isolated after the last blood withdrawal. The drug concentrations in blood and major organs were measured via optical imaging. The WB-PBPK model was constructed using known physiological values including the volumes of major organs and blood/lymphatic flow. The NONMEM software (version 7.3) was used to determine tissue partition coefficients. Using the WB-PBPK model, a clinical trial simulation was performed with reference to human physiological values acquired from the literature. The simulated human PK was then compared with the actual PK observed in the previous study in which healthy male subjects received 6 mg/kg trastuzumab (Herceptin®) via intravenous route. The ratio of the simulated versus observed area under the concentration-time curve was 1.02 and that of maximal concentration was 0.72. The current study describes the potential synergistic applications of WB-PBPK and optical imaging in human PK prediction based on preclinical data obtained in early-stage drug development.
School of Mathematical and Computer Sciences
Heriot-Watt University
Edinburgh, EH14 4AS, UK
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