Pharmaceutical Research[JOURNAL]
Sun
200 papers
Regional Nasal Drug Deposition in Pediatric vs. Adult Models: In vitro Insights into User Technique and Breathing Patterns Sensitivity.
Hejazi M, Owen X, Edwards DJ … +8 more , Hindle M, Longest W, Schuman T, Walenga R, Chopski S, Kaviratna A, Newman B, Golshahi L
Pharm Res · 2026 Apr · PMID 41840267 · Full text
PURPOSE: Although in vitro studies recommended for establishing bioequivalence of locally acting nasal suspension products provide useful information regarding product performance, currently they are not designed to full...
PURPOSE: Although in vitro studies recommended for establishing bioequivalence of locally acting nasal suspension products provide useful information regarding product performance, currently they are not designed to fully account for variability introduced by patient-specific factors such as administration technique and breathing patterns. This limitation, combined with ethical and practical challenges in conducting pediatric clinical trials, creates a knowledge gap in understanding age-related differences in nasal drug delivery. METHODS: Anatomically realistic in vitro nasal models representing an average child and adult, in terms of total drug delivery to the sites of intended action posterior to nasal valve, were used to investigate the sensitivity of regional nasal drug deposition to variations in administration parameters and breathing conditions. Three breathing conditions were evaluated: breath hold, gentle sniffing, and vigorous sniffing. Administration parameters were varied using a Box-Behnken experimental design. RESULTS: Results showed that for commercially available nasal sprays, breathing patterns did not significantly affect drug delivery in the adult model, but extreme conditions (no breathing vs. vigorous sniffing) caused significant differences in anterior deposition in the child model. Differences between gentle sniffing and the other conditions were not statistically significant. The sensitivity analysis revealed that in vitro drug deposition patterns differ between child and adult nasal cavities under identical user parameter variations, and variable importance differed across nasal cavity regions. CONCLUSIONS: This study demonstrates the potential for targeted drug delivery by enabling the identification of the optimal administration parameters to achieve desired outcomes, such as enhanced drug deposition in specific nasal regions.
Age-Appropriateness of Oral Formulations Authorized for Pediatrics.
Weiler HR, Le LM, Fadda HM
Pharm Res · 2026 Mar · PMID 41840266 · Publisher ↗
PURPOSE: Pediatric global regulatory initiatives have come into place to address gaps in evidence-based medicine for children. Objective of this study was to assess age-appropriateness of oral pediatric formulations auth...
PURPOSE: Pediatric global regulatory initiatives have come into place to address gaps in evidence-based medicine for children. Objective of this study was to assess age-appropriateness of oral pediatric formulations authorized through the U.S. Food and Drug Administration's Pediatric Rule, Best Pharmaceuticals for Children Act and Pediatric Research Equity Act. METHODS: Formulation age-appropriateness was assessed using an adapted version of the World Health Organization pediatric quality product profile assessment tool. Evaluations encompassed four product attributes across pediatric subgroups: dose and dose flexibility, patient acceptability, excipient safety and administration considerations. Each attribute was scored on a three-point scale: 1 indicating high risk, 2 moderate risk, and 3 low risk. RESULTS: 214 oral formulations were evaluated. Age-appropriateness remains a concern; with less than one third of authorized oral formulations for neonates meeting criteria for age-appropriateness, in comparison to 66% for older pediatric populations. Tablets and capsules constitute 58% of formulations indicated for pediatrics, despite tablet sizes often being unsuitable for younger children. Liquid formulations for neonates present a significant oversight, as they frequently contain excipients of potential concern. As children age, swallowability of the dosage form appears to be deprioritized, with only 37% of unswallowable large size tablets for school-aged children, available in alternative dosage forms such as granules, or solids for dispersions. CONCLUSIONS: Significant formulation gaps persist in pediatric medicine, with acceptability and excipient safety presenting highest risk. The potential of non-conventional solid dosage forms remains underutilized, underscoring the need for regulatory frameworks to support development of pediatric medicines.
Impact of Postnatal Changes in Creatinine Levels on Plasma Gentamicin Concentrations in Neonates.
Tsushita H, Tanaka R, Inoue M … +6 more , Tatsuta R, Kishimoto S, Maeda T, Yoshikawa N, Ihara K, Itoh H
Pharm Res · 2026 Apr · PMID 41840265 · Full text
OBJECTIVE: Gentamicin (GM), a renally eliminated drug, is widely used to treat infections in neonates. Neonatal dosing is typically based on body weight and postmenstrual age (PMA). Although serum creatinine (Cr) is the...
OBJECTIVE: Gentamicin (GM), a renally eliminated drug, is widely used to treat infections in neonates. Neonatal dosing is typically based on body weight and postmenstrual age (PMA). Although serum creatinine (Cr) is the gold standard for evaluating renal function, Cr at birth may reflect both neonatal and maternal levels. This study aimed to identify determinants of trough GM concentrations, focusing on renal function parameters. METHODS: This retrospective, single-center, observational study included 78 neonates who started intravenous GM on postnatal days 0-1. Dosing regimen was stratified by birth weight: < 1200 g (n = 22), 5 mg/kg every 48 h; 1200-1999 g (n = 10), 4 mg/kg every 36 h; ≥ 2000 g (n = 46), 4 mg/kg every 24 h. Dose-normalized trough concentration (C/D) was calculated by dividing trough GM concentration by 24-h equivalent GM dose. Cr level measured at birth (Cr) and the first Cr level measured after GM initiation (Cr) were analyzed. RESULTS: In neonates < 1200 g and 1200-1999 g, Cr was significantly higher than Cr. C/D correlated significantly with PMA, Cr, and urine output, but not with Cr. Multiple regression analysis incorporating variables measured at birth (Cr and PMA) as covariates identified PMA as the only significant predictor of C/D. When variables available following GM initiation (Cr, PMA, Cr, urine output, and ibuprofen co-administration) were included as covariates, Cr emerged as the sole independent factor predicting C/D. CONCLUSION: PMA-based dosing appears to be preferable for determining initial GM dosing on postnatal day 0, whereas Cr-based renal function parameters may be more effective for guiding maintenance dosing.
Advancing Sterilization of Medical Implant Polymers: Novel Low-Temperature Deep-vacuum Vaporized HO Technology Surpasses Current Methods.
Nallathambi A, Chiesa E, Rosalia M … +4 more , Bruni G, Tamborini A, Crotti S, Genta I
Pharm Res · 2026 Apr · PMID 41840264 · Full text
PURPOSE: To develop a deep vacuum vaporized hydrogen peroxide (VH₂O₂) sterilization method capable of preserving the structural and functional integrity of biocompatible polymeric electrospun mats usable as advanced bioc...
PURPOSE: To develop a deep vacuum vaporized hydrogen peroxide (VH₂O₂) sterilization method capable of preserving the structural and functional integrity of biocompatible polymeric electrospun mats usable as advanced biocompatible medical implants and devices, with particular focus on the influence of vacuum level during treatment. METHODOLOGY: Electrospun mats made of poly-L-lactide-co-glycolide (PLGA), poly-L-lactide-co-ε-caprolactone (PLC), and thermoplastic polyurethane (TPU) were prepared and sterilized with VH₂O₂ under low-vacuum (LV, 10 mbar) and high-vacuum (HV, 2 mbar) conditions at 20-50 °C. Scanning electron microscopy (SEM) assessed fiber morphology and pore area, contact angle measurements evaluated surface wettability, Fourier-transform infrared spectroscopy (FTIR) detected chemical modifications, and gel permeation chromatography (GPC) analyzed weight-average molecular weight (Mw) and polydispersity index (PI). RESULTS: HV sterilization caused fiber compression across all polymers, increasing fiber diameter and reducing pore area. LV sterilization conditions prevented morphological changes in TPU and PLC, while PLGA remained sensitive under both vacuum levels. PLGA also exhibited increased hydrophilicity due to surface reorganization and transition from Cassie-Baxter to Wenzel-type wetting. FTIR showed no chemical changes, and GPC confirmed stable Mw and PI for all tested materials (PLGA: 25-27 kDa; PLC: 16-18 kDa; TPU: 23-25 kDa). CONCLUSION: Deep vacuum VH₂O₂ sterilization at room temperature is feasible, with vacuum level critical for maintaining scaffold integrity. LV sterilization effectively preserves morphology in PLC and TPU, while HV is also reliable for less sensitive polymers. Chemical and molecular properties remain unaffected, supporting VH₂O₂ sterilization as a gentle and effective promising method for implantable textile-based biomaterials.
Advances and Further Comparison of Software Tools for Fisher Information Matrix-Based Design Evaluation in Pharmacometrics.
Fayette L, Brendel K, Mentré F
Pharm Res · 2026 Apr · PMID 41840263 · Publisher ↗
AIM: Before conducting a clinical study involving pharmacometrics analyses using Non-Linear Mixed Effects Models (NLMEM), study design can be evaluated by computing the Fisher Information Matrix (FIM) via first-order mod...
AIM: Before conducting a clinical study involving pharmacometrics analyses using Non-Linear Mixed Effects Models (NLMEM), study design can be evaluated by computing the Fisher Information Matrix (FIM) via first-order model linearisation. A 2015 study showed that several dedicated tools provided consistent results, with block-diagonal FIM approximations aligning more closely with clinical trial simulations (CTS) than full matrix approaches. Since, some tools have evolved with new features like covariate modelling and inter-occasion variability (IOV), while others have been newly developed within software initially created for parameter estimation in NLMEM. Our first aim is to compare predictions from both older (PFIM, PopED) and newer tools (NONMEM$Design, MlxDesignEval and Pumas OptimalDesign), and secondly to compare available estimation software to derived empirical predictions from CTS. METHODS: The 2015 examples, involving an analytical pharmacokinetic (PK) model and a pharmacokinetic/pharmacodynamic model described by ODE, and new cases including a PK model with covariates and a cross over design with IOV were implemented in the five software. Predicted relative standard error (RSE) and D-criterion were computed. Empirical RSE and D-criterion were estimated from CTS with the different estimation tools. RESULTS: For a given FIM approximation, both RSE and D-criterion were consistent across evaluation software. Results derived from block diagonal FIM were more aligned with CTS, for which results were comparable across estimation tools. Predictions were especially reliable for covariate effects and appropriately reflected the order of magnitude of IOV variances. CONCLUSION: Users can benefit from both reliable uncertainty prediction and parameter estimation regardless of their preferred software.
Evaluation of sNfL as a Biomarker for Paclitaxel-Induced Peripheral Neurotoxicity Through an Integrated PKPD Model.
Ibrahim EIK, Girdenyté M, Hu Y … +11 more , Di Cesare Mannelli L, Balayssac D, Busserolles J, Theil D, Roussignol G, Perrault O, Le Berre N, Chanut F, Slaoui M, Loryan I, Friberg LE
Pharm Res · 2026 Apr · PMID 41826588 · Full text
BACKGROUND: Serum neurofilament light chain (sNfL), a biomarker of axonal damage, has shown promise in clinical studies for monitoring paclitaxel-induced peripheral neurotoxicity (PIPN). The latter involves pathological...
BACKGROUND: Serum neurofilament light chain (sNfL), a biomarker of axonal damage, has shown promise in clinical studies for monitoring paclitaxel-induced peripheral neurotoxicity (PIPN). The latter involves pathological changes in PIPN sites such as the dorsal root ganglia, peripheral nerves, and brain. However, the mechanistic link between paclitaxel and NfL concentrations in these tissues remains poorly understood, necessitating preclinical investigation. METHODS: We developed a semi-mechanistic pharmacokinetic-pharmacodynamic model to characterize: (i) total and unbound paclitaxel concentrations in plasma, as well as in extracellular and intracellular compartments of PIPN sites; (ii) paclitaxel-tubulin complex formation; and (iii) NfL kinetics. The model was built using de novo-generated and previously reported data from rodents. RESULTS: Plasma pharmacokinetics of paclitaxel was captured using a two-compartment model, including Cremophor EL trapping and nonlinear tissue distribution. Paclitaxel pharmacokinetics in PIPN sites incorporated paclitaxel transport across the blood-to-PIPN sites barriers and paclitaxel-tubulin binding, described by capacity-limited kinetics with increased tubulin binding upon repeated plasma exposure. NfL kinetics in serum and cerebrospinal fluid were described using turnover models, with NfL leakage driven by paclitaxel-tubulin complex formation in PIPN sites. The model robustly predicted paclitaxel exposure across multiple doses and studies. While NfL predictions aligned with single-dose data, the model slightly underpredicted sNfL levels in an external validation dataset after repeated dosing of paclitaxel at 15 mg/kg, suggesting additional mechanisms may be involved. CONCLUSIONS: Overall, the model successfully described the relationship between paclitaxel exposure and sNfL kinetics, offering a model-based framework for translational studies.
Population Pharmacokinetics of Voriconazole in Patients Receiving Extracorporeal Membrane Oxygenation.
Meng M, Yao Y, Huang Z … +5 more , Tan C, Chen Y, Xiao Y, Xiang S, Chen X
Pharm Res · 2026 Mar · PMID 41826587 · Publisher ↗
PURPOSE: To characterize the population pharmacokinetics (PPK) of voriconazole (VRC) in critically ill patients receiving extracorporeal membrane oxygenation (ECMO) and to identify factors contributing to pharmacokinetic...
PURPOSE: To characterize the population pharmacokinetics (PPK) of voriconazole (VRC) in critically ill patients receiving extracorporeal membrane oxygenation (ECMO) and to identify factors contributing to pharmacokinetic (PK) variability to providing a scientific basis for the individualized and rational use of VRC. METHODS: Blood samples were collected 30 min before dosing and at 0.5, 1, 2, 4, 6 and 8 h after dosing. A PPK model was developed using nonlinear mixed effects modeling (NONMEM) and evaluated for stability and predictive performance. Based on the final model, Monte Carlo Simulations (MCS) were conducted to estimate the probability of target attainment (PTA) of therapeutic concentrations under various dosing regimens across different aspartate aminotransferase (AST) stratifications, with the aim of identifying optimal dosing strategies. RESULTS: This prospective study enrolled 30 critically ill adult patients receiving ECMO support, from whom a total of 183 VRC plasma concentration were obtained. VRC PK was adequately described by a two-compartment model. AST was identified as a significant covariate influencing VRC clearance. Model evaluation demonstrated that the final model exhibited robust stability and predictive performance. MCS results indicated that patients with AST levels > 65 U/L could achieve the therapeutic target range of VRC trough concentrations with lower loading and maintenance doses. CONCLUSIONS: This study provides quantitative evidence to support the optimization of VRC dosing in critically ill adult patients receiving ECMO support. The findings may improve the precision of individualized antifungal therapy and serve as a reference for the development of future dosing strategies in this population.
The Effect of Pharmaceutical Excipients on Protein Chemical Degradation Through Deamidation and Isomerization.
Ramm I, Diehl C, Västberg A … +4 more , Hjalte J, Schagerlöf H, Wahlgren M, Nilsson L
Pharm Res · 2026 Apr · PMID 41826586 · Full text
INTRODUCTION: Therapeutic proteins are crucial in the treatment of a wide range of diseases. However, the proteins are sensitive to chemical degradation reactions, particularly deamidation and isomerization, which can co...
INTRODUCTION: Therapeutic proteins are crucial in the treatment of a wide range of diseases. However, the proteins are sensitive to chemical degradation reactions, particularly deamidation and isomerization, which can compromise efficacy and safety. Formulation excipients, such as sugars and non-ionic surfactants, are commonly used to enhance stability, yet their effects on chemical degradation remain insufficiently understood. METHODS: This study investigates how fructose, sucrose, melezitose, and the non-ionic surfactants polysorbate 80 and DDM (n-Dodecyl-β-D-maltoside) affect the structure and chemical stability of the affibody GA-Z, a protein prone to deamidation and isomerization. Chemical degradation and conformational changes were characterized using peptide fingerprinting, Liquid Chromatography-Mass Spectrometry, Titration fluorescence spectroscopy, two-dimensional Nuclear Magnetic Resonance spectroscopy, and Differential Scanning Calorimetry. RESULTS: All three sugars lowered chemical degradation by stabilizing the folded state of the z-domain and inducing minor structural changes in the albumin-binding domain, thereby lowering the propensity for deamidation and isomerization. Polysorbate 80 showed minimal impact on both degradation and protein structure. In contrast, DDM increased deamidation and isomerization due to surfactant-protein interactions, resulting in structural changes. CONCLUSION: These results demonstrate how excipient-induced structural changes affect chemical degradation of proteins in liquid formulations. This study contributes to the understanding and design of more effective formulations for therapeutic proteins, enhancing their stability and safety.
Investigating the Potential of High Permeation Vesicle-Based Methotrexate Gel for Localized Therapy in Rheumatoid Arthritis.
Kumar K, Akhtar J, Dighe S … +3 more , Yadav V, Ansari MM, Jain S
Pharm Res · 2026 May · PMID 41826585 · Publisher ↗
OBJECTIVE: Methotrexate (MTX) is a clinically approved, potent disease-modifying antirheumatic drug used to manage rheumatoid arthritis. However, its systemic administration is associated with severe toxicity and bioavai...
OBJECTIVE: Methotrexate (MTX) is a clinically approved, potent disease-modifying antirheumatic drug used to manage rheumatoid arthritis. However, its systemic administration is associated with severe toxicity and bioavailability challenges. Localized MTX therapy holds significant potential by enabling site-specific action while diminishing the risk of systemic toxicity. METHODS: In this study, we investigated the therapeutic potential of systematically designed methotrexate-loaded high permeation vesicles (MTX-HPVs) in rheumatoid arthritis. HPVs comprise biocompatible phospholipids and a synergistic combination of permeation enhancers optimized using a quality-by-design approach. The thin-film hydration technique was used to formulate MTX-HPVs. Furthermore, MTX-HPVs were integrated into the Carbopol® 934P NF gelling matrix for ease of application and prolonged retention. RESULTS: The optimized MTX-HPVs exhibited optimal quality attributes with a sustained-release pattern for up to 48 h. Morphological assessment revealed a spherical shape of MTX-HPVs. The prepared MTX-HPVs loaded gel displayed enhanced flux (~ 6.9-fold) and permeation (~ 3.5-fold) compared to the free MTX gel. Furthermore, increased cellular internalization and reduced IC (0.57 ± 0.03 µg/mL) of the formulation in macrophage cells indicated the improved therapeutic potential of HPV-based therapy. In vivo studies revealed that MTX-HPVs gel significantly reduced inflammation and exhibited superior safety compared to free MTX gel. Radiographic imaging corroborated the enhanced efficacy and joint-restorative action of this formulation. CONCLUSION: The MTX-HPVs gel substantially enhanced the therapeutic efficacy of MTX, demonstrating HPVs as a promising carrier system for localized methotrexate therapy.
VEGF and Neurotrophins Interaction in the Retinal Neurovascular Unit Homeostasis: A Target for Ocular Disease Treatment and Management.
Di Criscio A, Rosso P, Fico E … +4 more , Iannetta D, Marenco M, Lambiase A, Tirassa P
Pharm Res · 2026 Mar · PMID 41814099 · Full text
Emerging evidence underscores the central role of the retinal neurovascular unit (RNVU) in the pathogenesis of major retinal disorders, including diabetic retinopathy, age-related macular degeneration, and glaucoma. Trad...
Emerging evidence underscores the central role of the retinal neurovascular unit (RNVU) in the pathogenesis of major retinal disorders, including diabetic retinopathy, age-related macular degeneration, and glaucoma. Traditionally considered as primarily vascular diseases, these conditions are now increasingly recognized to involve early neurodegenerative processes that may precede vascular dysfunction. Although anti-VEGF therapies have revolutionized the treatment of neovascular retinal diseases, long-term VEGF inhibition has been associated with adverse effects, including retinal atrophy and diminished neuroprotection, underscoring the need for more targeted strategies. Recent studies have highlighted the differential roles of VEGF-A splice isoforms, particularly the pro-angiogenic VEGF-Axxxa and the anti-angiogenic VEGF-Axxxb, in maintaining RNVU homeostasis and contributing to disease progression. In parallel, neurotrophins such as nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) have demonstrated the ability to exert neuroprotective, anti-inflammatory, and vasomodulatory effects, partly through modulation of VEGF-A signaling. Notably, we have recently demonstrated that NGF modulates VEGF-A isoform expression and VEGFR-2 levels in diabetic retinas, further supporting the hypothesis of a functional cross-talk between neurotrophins and angiogenic pathways. Based on this evidence, a new model is proposed, in which NGF and BDNF interact bidirectionally with VEGF-A to preserve RNVU integrity. This integrated therapeutic perspective, combining neurotrophic support with selective modulation of VEGF-A isoforms, may enhance treatment efficacy, reduce long-term side effects, and minimize the burden of care in chronic retinal neurodegenerative diseases.
Prediction of Large-Scale Adsorption Process Parameters Using Two-Mechanism Langmuir Constants.
Assaf Z, Wurster DE
Pharm Res · 2026 Mar · PMID 41807763 · Publisher ↗
Fitting the Langmuir-Like Equation to adsorption isotherms provides capacity and affinity constants which allow fundamental understanding of adsorption but are not helpful for large-scale processes where the relation to...
Fitting the Langmuir-Like Equation to adsorption isotherms provides capacity and affinity constants which allow fundamental understanding of adsorption but are not helpful for large-scale processes where the relation to the initial concentrations is most needed. In this paper, a novel equation is derived that allows the achievable final concentrations to be predicted based on any user-defined starting concentration, adsorbent loading, or solution volume. The equation accounts for both site-specific bonding and hydrophobic bonding and provides versatile plots while being simple to use in a spreadsheet format. The adsorption of barbituric acid, fluoxetine, phenobarbital, and procaine to 4 types of commercial activated carbons at different pH values, temperatures, buffers, and cosolvents is studied as an example. For instance, it was shown that for a 0.8 mg/mL initial solution of phenobarbital, a 20% increase of carbon mass will only result in a 5% decrease in final concentration. At 0.2 mg/mL initial phenobarbital concentration, no considerable improvement in adsorption can be observed for a loading mass to solution ratio > 0.75 mg adsorbent /mL solution. For adsorbates with a high extent of hydrophobic bonding, great reductions in concentrations, approaching zero, can be achieved even at high initial concentrations. The derived equation is free of assumptions, and is not specific to the systems studied.
Continuous Electrospraying of Inhalable Biological Drug Products: Novel Formulation and Manufacturing Technology.
Najafian S, Lei Y, Yang DH … +4 more , Mehta T, Luo Y, Stuber M, Chaudhuri B
Pharm Res · 2026 Mar · PMID 41807762 · Publisher ↗
PURPOSE: Traditional pharmaceutical manufacturing largely depends on batch processes that contribute to equipment downtime, higher carbon footprint, contamination risks, and elevated operational costs. Additionally, the...
PURPOSE: Traditional pharmaceutical manufacturing largely depends on batch processes that contribute to equipment downtime, higher carbon footprint, contamination risks, and elevated operational costs. Additionally, the oral delivery of biologics remains challenging due to degradation and poor gastrointestinal absorption. This study aims to develop and evaluate a continuous manufacturing platform based on electrospraying to produce a dry powder inhalation formulation of biologics. METHOD: A continuous electrospraying system was designed to generate core-shell structured microparticles composed of biodegradable PLGA encapsulating bovine serum albumin (BSA) and insulin. The microparticles were subsequently adhered to lactose carriers using a custom-built conveyor-belt-assisted collection setup. Particle morphology was examined via scanning electron microscopy, while protein distribution was assessed using fluorescence microscopy. Protein secondary structure and conformational stability were analyzed using FTIR and circular dichroism (CD), respectively. Encapsulation efficiency and in vitro release were quantified using UV-Vis spectroscopy. RESULTS: The electrospraying process successfully produced PLGA core-shell microparticles approximately 4-20 µm in diameter, uniformly attached to lactose carrier particles. Fluorescence analysis confirmed relatively uniform protein distribution within the particles. FTIR and CD measurements indicated that the protein structure remained nearly identical to the native form, demonstrating minimal structural alteration during processing. The formulation achieved an encapsulation efficiency of 78.16%, with nearly 72% cumulative protein release within 12 h. CONCLUSION: The developed electrospraying-based continuous manufacturing platform demonstrates strong potential as a scalable and versatile approach for producing stable, biologic-loaded DPI formulations. Its ability to encapsulate sensitive proteins while preserving structural stability highlights its promise for future inhalable biologic delivery applications.
Development and Validation of an Efficient In vitro Bioassay for Potency Estimation of Recombinant Human Erythropoietin Formulations.
Garg N, Gupta P, Kamal CM
Pharm Res · 2026 Apr · PMID 41792507 · Publisher ↗
PURPOSE: Recombinant human erythropoietin (rHuEPO) is regarded as a life-saving biotherapeutic in case of acute disease associated anemia. Several commercially available rHuEPO drug products are present in the market for...
PURPOSE: Recombinant human erythropoietin (rHuEPO) is regarded as a life-saving biotherapeutic in case of acute disease associated anemia. Several commercially available rHuEPO drug products are present in the market for patient consumption. Since manufacturers use different technologies to produce recombinant drugs, their quality, safety and efficacy becomes imperative as compared with the innovator. Compendial guidelines worldwide have laid down regulations and testing methodologies to ascertain the potency of biomolecular drugs. The current method for potency estimation of rHuEPO samples is animal-based, lengthy, tedious, ethically restrained and shows high variability. The current study employs a rather simple cell-based method that can be used as an alternative to the in vivo methods. METHODS: A cell proliferation method, with absorbance-based end point, was developed on UT-7 cell line for estimating the potency of rHuEPO products. The developed UT-7 cell line method was validated according to USP and ICH guidelines. The utility of method was further confirmed on different batches of rHuEPO drug products to determine their potency values. These batches simultaneously were analyzed by the existing in vivo method as cross validation studies. RESULTS: The results showed high level of reproducibility, precision, accuracy and range. The comparative studies of in vitro with in vivo methods depicted the potency of the batches by both the methods were in the acceptable range of 80-125% CONCLUSION: Though, this method doesn't account for the additional effects of a closed in vivo system, yet it presents itself as a simple and sensitive method that can be used for regular quality testing of rHuEPO formulations.
REAL208, a Liposome-based Formulation of Phosphodiesterase Inhibitor, Protects Acute Kidney Deficiency in Mice.
Tsai JC, Lin YW, Huang CY … +6 more , Shih CM, Wu YT, Kurosaki T, Inoue Y, Du YC, Lin FY
Pharm Res · 2026 Apr · PMID 41792506 · Full text
OBJECTIVE: Acute kidney injury (AKI) is a life-threatening condition characterized by inflammation, oxidative stress, and tubular cell apoptosis for which no specific pharmacological therapy currently exists. Dipyridamol...
OBJECTIVE: Acute kidney injury (AKI) is a life-threatening condition characterized by inflammation, oxidative stress, and tubular cell apoptosis for which no specific pharmacological therapy currently exists. Dipyridamole, a phosphodiesterase inhibitor with antiplatelet, anti-inflammatory, and cytoprotective properties, has shown renoprotective potential; however, its clinical use in AKI is limited by pharmacokinetic constraints. METHODS AND RESULTS: To address this issue, we developed REAL208, a liposome-encapsulated dipyridamole formulation and evaluated its safety and therapeutic efficacy in experimental AKI models. Safety testing in C57BL/6 mice demonstrated that REAL208 was well tolerated at doses up to 50 mg/kg, without evidence of hepatic or renal toxicity. In lipopolysaccharide (LPS)- and ischemia/reperfusion (I/R)-induced AKI models, intravenous administration of REAL208 (2.5-10 mg/kg) significantly improved survival, reduced serum blood urea nitrogen and creatinine levels, and restored glomerular filtration rate compared to that observed in the untreated controls. Histological analyses revealed preserved tubular architecture, reduced neutrophil and macrophage infiltration, and modulation of the expression of injury markers, including KIM-1 and PPARγ. Immunohistochemistry further showed attenuation of Nrf2 and HO-1 upregulation, indicating suppression of oxidative stress. In vitro, REAL208 protected HK-2 proximal tubular cells against LPS- or hypoxia/reperfusion-induced injury, maintained mitochondrial membrane potential, enhanced oxygen consumption rates, and reduced inflammatory signaling (nuclear p65 and Bcl-2). CONCLUSIONS: These findings demonstrated that liposomal dipyridamole exerts robust renoprotective effects by preserving mitochondrial function, suppressing oxidative stress, and attenuating inflammation. REAL208 represents a promising therapeutic strategy for AKI and warrants further translational and clinical investigations.
Fresh 3D Printing of Spanlastics Hydrogel for Drug Delivery Applications In Vitro.
Doe E, Alabi A, Chakka LRJ … +1 more , Maniruzzaman M
Pharm Res · 2026 Mar · PMID 41792505 · Publisher ↗
PURPOSE: To develop a localized delivery implant by integrating doxorubicin loaded spanlastic vesicles within Freeform Reversible Embedding of Suspended Hydrogels (FRESH) printed alginate constructs. METHODS: Spanlastics...
PURPOSE: To develop a localized delivery implant by integrating doxorubicin loaded spanlastic vesicles within Freeform Reversible Embedding of Suspended Hydrogels (FRESH) printed alginate constructs. METHODS: Spanlastics composed of Sorbitan Monostearate (Span60) and an edge activator, Polyethylene sorbitol ester (Tween 80), were prepared by ethanolic injection. Plain and drug-loaded spanlastics were characterized for their physicochemical properties. Vesicles were incorporated into 3D printed sodium alginate hydrogels in 'FRESH' bioprinting process to promote the sustained drug release of doxorubicin which was assessed using dialysis membrane for drug release. In vitro uptake and cytotoxicity were evaluated in MCF7 breast cancer cells. RESULTS: Optimized formulations produced vesicles of approximately 200 to 300 nm with moderate encapsulation efficiency (33 to 44%) and stability during hydrogel incorporation and printing. Printed depots provided sustained doxorubicin release relative to suspension and reduced MCF7 viability, with preferential intracellular and nuclear localization consistent with doxorubicin activity. CONCLUSION: Spanlastic-loaded FRESH printed alginate implants combine vesicle-mediated cellular delivery with matrix-governed sustained release, supporting their potential as a localized chemotherapy depot for further in vivo validation.
Integrated Experimental and Molecular Modeling Techniques to Investigate the Buffer Effects on Glucagon Stability.
Xu ZT, Li B, Deng J … +3 more , Lv JY, Ouyang D, Fang WJ
Pharm Res · 2026 Mar · PMID 41792504 · Publisher ↗
PURPOSES: Peptide drugs are a vital category of biologics. However, unlike the well-studied effects of buffers on small-molecule or macromolecule drugs, the understanding of selecting appropriate buffers for peptide form...
PURPOSES: Peptide drugs are a vital category of biologics. However, unlike the well-studied effects of buffers on small-molecule or macromolecule drugs, the understanding of selecting appropriate buffers for peptide formulations and their specific impacts remains insufficient. This study aimed to systematically evaluate how different buffers affect the stability of the model peptide, glucagon, and to investigate the underlying mechanisms at the molecular level. METHODS: This work employed glucagon as a model peptide and integrated experimental techniques with computational approaches. Experimental techniques included reverse-phase high-performance liquid chromatography (RP-HPLC), Thioflavin T fluorescence assays, and circular dichroism (CD). Molecular dynamics (MD) simulations were further conducted, utilizing models with different buffer systems and varying peptide counts, to examine the interaction mechanisms between buffers and glucagon from multiple perspectives. RESULTS: Compared with other buffers, citrate exerted a unique and significant impact on glucagon stability. RP-HPLC demonstrated its destabilizing effect on the structure, while CD and MD simulations confirmed its role in preserving glucagon's α-helical structure, attributed to its highest binding energy. However, this strong binding reduced the ζ-potential, compromised colloidal stability, and ultimately promoted aggregation/precipitation. MD simulations further showed that citrate anions formed a denser solvation shell around glucagon, driving oligomerization and aggregation. CONCLUSIONS: This work uncovers the dual role of citrate buffer on glucagon stability: maintaining local secondary structure while disrupting overall colloidal stability. It provides molecular insights into peptide-excipient interactions and offers valuable guidance for optimizing formulations for other peptide drugs.
Impurity Characterization Across Drug Development Stages: Analytical Methodologies and Regulatory Perspectives.
Ramamoorthy S
Pharm Res · 2026 Mar · PMID 41776153 · Publisher ↗
The rigorous identification, quantification, and control of impurities are fundamental to ensuring the safety, efficacy, and quality of pharmaceutical products throughout the drug development lifecycle. As unavoidable by...
The rigorous identification, quantification, and control of impurities are fundamental to ensuring the safety, efficacy, and quality of pharmaceutical products throughout the drug development lifecycle. As unavoidable byproducts of complex synthesis and manufacturing processes, impurities span a diverse range of compounds-including organic, inorganic, residual solvent, and genotoxic species-all of which require meticulous characterization using a suite of analytical techniques. This review synthesizes best practices for impurity classification, source mapping, and alignment of control strategies with global regulations (ICH Q3A/B, Q3C, Q3D, and M7), distinguishing phase-appropriate profiling, stress testing, and fully validated, stability-indicating methods. Advanced structural-elucidation tools (LC-MS/MS, HRMS, NMR) are surveyed alongside key strategies for lifecycle change management. Special emphasis is placed on mutagenic impurities, especially nitrosamines, highlighting how recent industry experience and regulatory actions have reshaped risk management and analytical sensitivity requirements. The discussion extends to emerging trends-green analytical chemistry, AI-driven prediction and interpretation, and real-time monitoring via Process Analytical Technology (PAT) and Real-Time Release Testing (RTRT)-that are redefining impurity control from reactive detection to proactive prevention. Collectively, this review provides scientists, regulators, and quality professionals with actionable frameworks for robust impurity management, facilitating compliance and accelerating pharmaceutical development.
Small Molecules to Elevate Rab7-GTPase Activity and Lower Cholesterol Accumulation in Niemann-Pick Type C Disease.
Nguyen MKL, Nikenich MR, Seifert K … +24 more , Pinkenburg C, Lai F, Walther HL, Hartmann M, Szulc A, Sparkes E, Chen S, Wikramanayake R, Bernaus-Esqué M, Liu Y, Tebar F, Serwetnyk M, Wenninger A, Patzke C, Blagg BSJ, Ashfeld BL, Groundwater PW, Hibbs DE, Hoy AJ, Rentero C, Enrich C, Cho AN, Du JJ, Grewal T
Pharm Res · 2026 Apr · PMID 41776152 · Full text
PURPOSE: Niemann-Pick type C (NPC) disease caused by mutations in cholesterol transporters NPC1 or NPC2 is characterized by cholesterol accumulation in late endosomes/lysosomes (LE/Lys). The activation of alternative cho...
PURPOSE: Niemann-Pick type C (NPC) disease caused by mutations in cholesterol transporters NPC1 or NPC2 is characterized by cholesterol accumulation in late endosomes/lysosomes (LE/Lys). The activation of alternative cholesterol export routes that can bypass NPC1/2 deficiency could provide therapeutic opportunities. We previously demonstrated that gene depletion of the Rab7-GTPase activating protein (GAP) TBC1D15, which hydrolyses active GTP-bound Rab7, led to elevated Rab7-GTP levels. This enabled cholesterol export from LE/Lys to reduce cholesterol accumulation in NPC1 mutant cells. Here we aimed to pharmacologically interfere with TBC1D15-mediated Rab7 inactivation to upregulate Rab7 activity and reduce cholesterol accumulation in NPC1 mutant models. METHODS: The protein structure of the GAP domain of human TBC1D15 in complex with human Rab7-GTP served to perform in silico drug screening and identify small molecules with potentially high TBC1D15 binding affinity. Rab-GTP pulldown assays and fluorescence microscopy analyzed the ability of drug candidates to elevate Rab7-GTP levels and reduce cholesterol accumulation. RESULTS: Four drug candidates reduced cholesterol accumulation in NPC1 mutant Chinese Hamster Ovary (CHO) M12 cells, NPC1 patient fibroblasts as well as differentiated SH-SY5Y neuronal cells and three-dimensional brain organoids treated with U18666A, a pharmacological NPC1 inhibitor. This was associated with elevated Rab7-GTP levels in drug-treated M12 and NPC1 patient fibroblasts. Moreover, drug candidates augmented 2-hydroxypropyl-β-cyclodextrin (HPβCD)-induced cholesterol removal from U18666A-treated SH-SY5Y cells. Notably, drug candidates did not negatively impact on cell viability or cause membrane damage. CONCLUSION: Advancing small molecules that can elevate Rab7-GTPase activity could provide opportunities to overcome cholesterol transport defects in NPC mutant cells and offer applications in other Rab7-related neurological diseases.
Solid-State Stability of Pentobarbital in Fast-Dissolving Suppositories: Implications for Dissolution Behavior and Pediatric Formulation Performance.
Freisz A, Dhifallah I, Chennell P … +8 more , Yessaad M, Wasiak M, Jouffret L, Petit E, Lambert C, Jouannet M, Sautou V, Bouattour Y
Pharm Res · 2026 Apr · PMID 41776151 · Publisher ↗
BACKGROUND: Safe and well-tolerated sedative formulations are essential in pediatric procedural care. Pentobarbital, a short-acting barbiturate, remains widely used for pediatric sedation. However, its oral administratio...
BACKGROUND: Safe and well-tolerated sedative formulations are essential in pediatric procedural care. Pentobarbital, a short-acting barbiturate, remains widely used for pediatric sedation. However, its oral administration is limited by poor palatability. Rectal administration therefore represents a relevant alternative, provided that formulation stability is ensured. This study evaluated the physicochemical and microbiological stability of fast-dissolving pentobarbital suppositories developed for pediatric use. METHODS: Suppositories containing 30 or 60 mg of pentobarbital sodium were formulated with two sodium hydroxide (NaOH) concentrations and stored under refrigerated (5°C) or frozen (-20°C) conditions for up to 180 days. Stability was assessed by macroscopic examination, pH measurement, dissolution testing, mechanical strength, softening time, and microbiological quality. Solid-state changes were analyzed using Raman spectroscopy and X-ray diffraction, and precipitation kinetics were modeled. RESULTS: Refrigerated storage led to progressive precipitation from day 90, associated with pH decrease and conversion of pentobarbital sodium to its free acid, resulting in incomplete dissolution. In contrast, frozen storage preserved pH, physical integrity, and complete dissolution, even after 30 days at 5°C following thawing. Spectroscopic and diffraction analyses confirmed recrystallization under refrigeration, whereas frozen formulations retained an amorphous structure. Increased NaOH content delayed, but did not prevent, instability at 5°C. All formulations complied with pharmacopeial mechanical and microbiological requirements. CONCLUSION: Storage temperature was the main determinant of stability, with freezing ensuring long-term preservation. NaOH buffering mitigated gelatin acidity and delayed drug conversion, thereby extending shelf life. These findings support further biopharmaceutical and clinical evaluation of pentobarbital suppositories for pediatric sedation.
Near UV and Visible Light-Induced Degradation of IgG2: Photo-Fenton Reaction in Citrate Buffer.
Subelzu N, Mozziconacci O, Schöneich C
Pharm Res · 2026 Apr · PMID 41776150 · Publisher ↗
PURPOSE: Evaluation of the stability of an IgG2 in citrate buffer, upon exposure to near-UV and visible light and in the presence of relevant trace amounts of iron. METHODS: We monitored the oxidation of amino acid resid...
PURPOSE: Evaluation of the stability of an IgG2 in citrate buffer, upon exposure to near-UV and visible light and in the presence of relevant trace amounts of iron. METHODS: We monitored the oxidation of amino acid residues, the formation of protein aggregates, fragments and charge variants by SDS-PAGE and two-dimensional gel electrophoresis (2-DIGE). Degradation products in individual gel spots were isolated and characterized by HPLC-MS/MS. RESULTS: We detected an increase in the formation of DOPA, protein aggregates, and fragments with increasing concentrations of added Fe. Light exposure resulted in the generation of more acidic products, evident from a shift of the protein pI detected by 2-DIGE. Oxidation products of Tyr, His, Cys, and Trp were detected. The addition of EDTA or DTPA showed a significant protection against degradation. CONCLUSIONS: IgG2 was significantly modified by the photo-Fenton reaction in citrate buffer. We demonstrated the oxidation of Tyr, His, Cys, and Trp residues, the formation of aggregation and degradation products, as well as the formation of different charge variants.
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