Pharmaceutics doi: 10.3390/pharmaceutics18091162

Authors: Blanca Vicente Hinojal Zazo José Germán Sánchez-Hernández Natalia Revilla Paulo Teixeira-da-Silva

Background: Subcutaneous infliximab (SC-IFX) represents a significant advance for patients with inflammatory bowel disease (IBD) due to the possibility of home self-administration. However, at this time, there are no clear recommendations for implementing model-informed precision dosing (MIPD) in accordance with real-world clinical knowledge. Objective: The aim of the present study is to develop a population pharmacokinetic (PopPK) model for SC-IFX. Methods: A multicenter retrospective observational study was performed in adult patients (both naïve patients and those switched from intravenous administration) diagnosed with IBD. PopPK analysis was performed using NONMEM with a nonlinear mixed-effects model approach. A stepwise covariate modeling approach was implemented to identify anthropometric and clinical factors influencing SC-IFX disposition. The bootstrap method was used for internal model validation. Results: A total of 250 serum concentration samples of IFX, corresponding to 45 patients, were analyzed. A one-compartment model with first-order absorption adequately described the data. The final model included total body weight and suspected immunogenicity as covariates of clearance (CL). Suspected immunogenicity was associated with a 70.0% increase in clearance, while each kg deviation from the median weight (73 kg) modified clearance by 0.888%/kg. Conclusions: A PopPK model for SC-IFX was successfully developed and validated using European real-world data. This exploratory model establishes a foundation for future MIPD applications, offering a structured approach to initiate or transition patients to SC-IFX therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091161

Authors: Mohammad Moein Sadeghi Souha Idoudi Helena M. Kelly Nashiru Billa

Biomaterials intended for wound healing are expected to reduce microbial load while supporting tissue repair in complex wound environments. Plant-mediated synthesis of silver nanoparticles (AgNPs) has attracted interest because plant extracts can provide phytochemicals that mediate nanoparticle reduction, capping, and stabilization. This review critically evaluates recent studies on plant-mediated AgNPs for wound-healing applications, focusing on botanical and extraction variables, synthesis conditions, physicochemical properties, formulation strategies, administered exposure, safety, and biological outcomes. Across the 34 included publications, 14 directly assessed a process-to-property relationship and only two assessed a process-to-performance relationship; none directly assessed a property-to-performance relationship or established a complete process-to-property-to-performance chain. The reviewed studies suggest that plant-mediated AgNPs may reduce microbial burden, modulate inflammation, mitigate oxidative stress, and promote cell migration, re-epithelialization, collagen remodeling, and angiogenesis. However, batch-to-batch variability, incomplete reporting of experimental details, over-reliance on ultraviolet–visible (UV–Vis) spectroscopy for optimization, inadequate evaluation of formulation stability, inconsistent dose reporting, and limited molecular validation of proposed mechanisms remain major factors limiting translation. By evaluating plant-mediated AgNP-based wound-healing systems through a process–property–performance framework, this review summarizes recent advances and highlights key considerations for improving reproducibility, comparability, and translational relevance.

Pharmaceutics doi: 10.3390/pharmaceutics18091160

Authors: Qilin Wang Tingyu Chen Hailong Dou Deng-Guang Yu

Background: Simultaneous controlled release of multiple therapeutics from a single dosage form is a major frontier in modern pharmaceutics and a key strategy for modernizing traditional Chinese medicine. Methods: Using finasteride (FIN) and a Jing-Zhu hybrid Tibetan (JZ) herbal medicine as model agents for prostatitis therapy, we developed a modified tri-fluid electrospinning process to fabricate drug-co-loaded Janus medicated nanofibers. Soluble polyvinylpyrrolidone (PVP) and insoluble ethylcellulose (EC) served as carrier matrices for the dual faces of the Janus architecture, encapsulating JZ herbal medicine and FIN, respectively. A custom-designed spinneret—comprising two parallel stainless steel tubes nested within a plastic sheath—was engineered to enable side-by-side fiber formation. Results: Scanning and transmission electron microscopy confirmed linear morphologies with a definitive side-by-side Janus structure. X-ray diffraction and Fourier Transform Infrared Spectroscopy revealed that all active ingredients were dispersed in an amorphous state, reflecting polymer–drug compatibility. Encapsulation efficiencies reached 96.34 ± 0.47% for FIN and 94.15 ± 0.48% for JZ herbal medicine. A newly devised water-droplet assay demonstrated that almost all the nanofibers exhibited the intended side-by-side configuration, as evidenced by the rapid dissolution of the PVP side. In vitro release studies showed an initial pulsatile burst of JZ herbal medicine followed by a sustained FIN release profile, as suggested by the single-drug-loaded Janus nanofibrous controls. Conclusions: The present Janus nanostructure system, fabricated via a facile co-shell solvent electrospinning process, has the potential to enable the concurrent yet asynchronous delivery of FIN and JZ herbal components within a single nano-dosage form. This conceptual advance expands the toolkit for designing combination nanomedicines, allowing independent modulation of release kinetics for individual drugs to maximize prospective joint efficacy.

Pharmaceutics doi: 10.3390/pharmaceutics18091159

Authors: Ahmed A. H. Abdellatif Hesham M. Tawfeek Mahmoud Zaki El-Readi Amani E. Alharbi Hamzah M. Maswadeh Mohammed A. Amin Safaa Y. Eid Abousree T. Ellethy Bassem Refaat Akhmed Aslam Waleed Mohammad Altowayan Mahmoud A. Younis

Background: Herein, we report on the development of a novel submicron-sized formulation that utilizes the somatostatin receptor agonist, octreotide (OCT), for the solubilization and targeted delivery of paclitaxel (PTX) and docetaxel (DTX) to the cancer cells. The formulation was prepared using a fast, single-step, and scalable method, and then, it was characterized in terms of physicochemical properties and pharmacological activities. Methods: Formulation analyses by Electrospray Ionization Mass Spectrometry (ESI-MS) and High-Performance Liquid Chromatography (HPLC) confirmed the successful formation of the formulation and the inclusion of all of its components. Results: Meanwhile, the developed formulation showed acceptable physicochemical properties with no incompatibilities among its components. In addition, the prepared formulation, OCT-PTX-DTX, demonstrated superior anticancer activities against colorectal carcinoma cell lines compared to the free drugs, with minimal effects on the normal cells. Furthermore, a synergistic anticancer activity of PTX and DTX was enabled by the novel formulation, leading to cell cycle arrest at low drug concentrations. Conclusions: Molecular-based mechanistic investigations verified that OCT-PTX-DTX is taken up by colorectal cancer cells via somatostatin receptors 1 and 2 (SSTR1 and SSTR2), followed by the upregulation of the tumor-suppressor p53/p21 cascade, and the induced cell cycle arrest via suppression of the cell cycle drivers CDK2 and Cyclin A. The formulation reported herein is promising for targeted and biotolerable anticancer chemotherapy compared to the commercially available taxane formulations based on surfactants or organic solvents.

Pharmaceutics doi: 10.3390/pharmaceutics18091158

Authors: Neha Tyagi Manish Rathi Susan Ensel Christopher C. Thornburg Tanja Grkovic Barry R. O’Keefe James C. Sacchettini Chendil Damodaran

Background/Objectives: The progression of prostate cancer to castration-resistant prostate cancer (CRPC) is often driven by constitutively active androgen receptor splice variants, such as AR-V7, which evade conventional androgen-deprivation therapies. This study aimed to develop a high-throughput, mechanism-based screening platform to identify natural-product-derived AR-V7 inhibitors from the NCI Program for Natural Product Discovery (NPNPD) library. Methods: A subset of prefractionated NPNPD samples was screened using a CRISPR-edited 22Rv1 cell line expressing endogenous AR-V7 fused to a HiBiT luminescent tag, enabling quantification of AR-V7 levels. Fractions that reduced the HiBiT signal were further evaluated in 22Rv1 and C4-2B CRPC cells and counter-screened in non-malignant RWPE-1 cells. Compounds demonstrating at least 90% inhibition in CRPC cells with no more than 10% toxicity in RWPE-1 cells underwent dose–response analysis, Western blotting, quantitative PCR, and subfractionation to isolate and characterize active constituents. Results: Of the 704 prefractionated samples, five fractions met stringent activity and selectivity criteria, with three consistently suppressing full-length androgen receptor (AR), AR-V7, and prostate-specific antigen (PSA). Primary screening results in HiBiT-22Rv1 cells were validated by secondary dose–response and Western blot assays, and hits were further prioritized to ensure toxicity remained at or below 10% in RWPE-1 cells. HiBiT-guided screening of 66 subfractions identified seven that robustly downregulated AR signaling, with reduced AR-V7 levels correlating with decreased CRPC cell viability. Chemical characterization revealed two active butanolides, isolitsealiicolide C and isolinderanolide B, which reduced AR and AR-V7 protein and mRNA levels, decreased PSA, downregulated Bcl-2, and induced cleaved PARP, consistent with apoptotic cell death in AR-positive CRPC models. Conclusions: This integrated high-throughput workflow efficiently identifies AR-V7–targeted natural products from complex libraries and highlights isolitsealiicolide C and isolinderanolide B as promising scaffolds for overcoming androgen receptor–driven resistance in CRPC.

Pharmaceutics doi: 10.3390/pharmaceutics18091157

Authors: Malamati Kourti Lefteris Zacharioudakis Annita Kolnagou Iliada Evripidou George J. Kontoghiorghes

Background: Artificial intelligence (AI) is transforming pharmacology and pharmacotherapy by enabling the integration of large, heterogeneous datasets to support precision and personalised medicine. However, the reliability of AI-assisted therapeutic decision-making depends fundamentally on the selection, quality, and interpretation of pharmacological and clinical input features or parameters. Current AI models frequently overlook the multidimensional complexity of drug- and patient-specific factors, limiting their clinical applicability and generalisability. Methods: This narrative review was prepared based on 45 years of experimental work and relevant published research in drug design, development, and clinical experience in iron chelation therapy and personalised treatment approaches. Additional literature was identified through targeted searches of PubMed and Scopus using terms related to AI, machine learning, pharmacology, pharmacotherapy, and personalised medicine. Key pharmacological and clinical input features relevant to AI-assisted personalised drug selection include physicochemical drug properties, absorption, distribution, metabolism, excretion and toxicity characteristics, route of administration, drug interactions, pharmacokinetics, pharmacodynamics, multi-omics data, therapeutic efficacy, diagnostic biomarkers, statistical validation, governance, and explainable AI. A conceptual framework for AI-assisted personalised drug selection is also proposed as an example. Results: It is suggested that reliable AI-assisted pharmacotherapy requires the integration of diverse, interdependent datasets and parameters describing drug characteristics, patient variability, clinical outcomes, and real-world evidence. The incorporation of pharmacogenomics, electronic health records, diagnostic imaging and profiling, and validated computational descriptors can improve prediction of drug efficacy, toxicity, interactions, and therapeutic response. Furthermore, robust model validation, data governance, cybersecurity, and continuous monitoring are identified as essential prerequisites for safe clinical implementation. The proposed conceptual framework illustrates how clinical admissibility filtering, model-based ranking, local attribution, and clinically supervised decision support may enhance personalised drug selection while maintaining human oversight. Conclusions: Artificial intelligence has considerable potential to improve drug selection and personalised pharmacotherapy. However, its success depends on comprehensive integration of pharmacological knowledge with high-quality clinical data, rigorous validation, and responsible governance. The multidimensional framework presented here provides a foundation for the potential development of clinically interpretable, reliable, and patient-centred AI systems capable of supporting safer and more effective precision medicine.

Pharmaceutics doi: 10.3390/pharmaceutics18091156

Authors: Weijian Zeng Duanyang Zhou Tianlong Wang Zhan-Lu Ma-Högemeier Song Cai Bingfeng Liu Chao Song Ling Guo Rihong Zhai Xun Song Zhendan He Yun Dong

Background: Oxidative stress-mediated neuronal injury is critically involved in the pathogenesis of neurodegenerative disorders, including Alzheimer’s disease and Parkinson’s disease. Manganese dioxide (MnO2), owing to its intrinsic reactive oxygen species (ROS)-scavenging capacity, has emerged as a promising neuroprotective candidate; however, the underlying molecular mechanisms remain insufficiently defined. Methods: Bovine serum albumin-templated MnO2 nanoparticles (BSA-MnO2 NPs) were synthesized, and their protective effects were evaluated in H2O2-treated PC12 cells. Results: BSA-MnO2 NPs significantly inhibited H2O2-induced reductions in cell viability, ROS overproduction, and mitochondrial membrane potential disruption. Mechanistically, H2O2 increased both LC3-II and p62 levels, indicating impaired autophagic flux. Activation of autophagy by serum starvation alleviated H2O2-induced injury, whereas chloroquine exacerbated cellular damage and abolished the protective effects of BSA-MnO2 NPs, suggesting that the restoration of autophagy contributes to BSA-MnO2 NPs-mediated neuroprotection. Further analysis showed that BSA-MnO2 NPs enhanced Akt phosphorylation, while LY294002, a PI3K inhibitor, suppressed Akt activation, disrupted autophagy regulation, and eliminated their neuroprotective effects. In contrast, chloroquine did not affect Akt phosphorylation, indicating that PI3K/Akt signaling acts upstream of autophagy regulation. Conclusions: Collectively, these findings demonstrate that BSA-MnO2 NPs protect PC12 cells against H2O2-induced oxidative injury by restoring autophagy through the PI3K/Akt signaling pathway, highlighting a potential role of BSA-MnO2 NPs in the treatment of oxidative-stress-related neurodegenerative disorders.

Pharmaceutics doi: 10.3390/pharmaceutics18091155

Authors: Li Wang Ning Yang Yina Sa Zhenghua Li Yanlong Qiao Yanxin Yang Lixue Chen Xiangyu Liu Lei Li

Background: combination chemotherapy employing agents with complementary mechanisms of action can enhance therapeutic efficacy and overcome resistance, but mismatched pharmacokinetics and systemic toxicity limit benefit. Methods: We designed reductant-responsive squalene prodrugs of doxorubicin (DOX) and exatecan (EXA) that co-assemble with DSPE-PEG2k into approximately 76 nm dual-prodrug nanoparticles (doxorubicin prodrug–exatecan prodrug nanoparticles, DP-EP NPs) at a predefined synergistic ratio 1:2. Results: The minimalist, drug-as-carrier formulation achieved high effective loadings with minimal leakage under normoxia, yet synchronously liberated both drugs in reductive, hypoxia-mimicking media (24 h release: DOX ~65% and EXA ~58% at 1 mM Na2S2O4 vs. ~8–11% at 0 mM Na2S2O4). In MCF-7 and 4T1 cells, the nanoparticles enhanced intracellular accumulation, intensified the disruption of DNA damage repair, as evidenced by increased colocalization of 53BP1 and enhanced γ-H2AX foci, and induced markers of immunogenic cell death, including increased CRT-associated cellular fluorescence and reduced intracellular HMGB1 staining. Furthermore, they partially reversed drug resistance in MCF-7/ADR cells. In rats, the formulation prolonged circulation and increased exposure (DOX t½ ~9×, AUC ~1.8×; EXA t½ ~5×, AUC ~14.9×) relative to the free-drug mixture. In 4T1 tumor-bearing mice, the nanoparticles produced the greatest tumor-growth inhibition with improved tolerability, supported by stable body weight and benign histopathology. Conclusions: These findings establish a scalable, excipient-lean platform that aligns pharmacokinetics with microenvironment-triggered pharmacodynamics to deliver synchronized Topo II/I inhibition for synergistic chemotherapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091154

Authors: Xiao-Ying Zhang Yue Li Jing Xu Yong-Jun Fang Xuan-Sheng Ding Feng Chen

Background/Objectives: Busulfan is a cornerstone conditioning agent for pediatric hematopoietic stem cell transplantation (HSCT), yet its narrow therapeutic index and substantial pharmacokinetic (PK) variability complicate dosing. This review synthesizes current evidence on pediatric busulfan PK, exposure-guided dosing, and the transition from conventional therapeutic drug monitoring (TDM) to model-informed precision dosing (MIPD). Methods: We conducted a focused narrative search of PubMed, Web of Science, and ScienceDirect from database inception to August 2026 and synthesized relevant literature on pediatric busulfan PK, exposure–response relationships, TDM, population PK modeling, Bayesian methods, and emerging quantitative approaches. Results: Body size is the primary determinant of busulfan clearance in children, while maturation, underlying disease, concomitant medications, and pharmacogenetic variation may contribute additional variability, although their effects are inconsistent across populations. Busulfan exposure is associated with HSCT outcomes and toxicity; however, reported targets differ by dosing regimen, conditioning intensity, disease category, and exposure metric. Reliable exposure estimation requires accurate sampling records and validated bioanalysis, while limited sampling strategies may improve clinical feasibility. Integrating population PK models with Bayesian estimation enables individualized dose adjustment, although routine implementation requires external validation and compatibility with local workflows. Emerging approaches, including physiologically based PK, PK/PD, and machine learning models, show promise but lack sufficient clinical validation for routine use. Conclusions: TDM remains the foundation of pediatric busulfan precision dosing, while MIPD provides a framework for integrating patient characteristics, measured concentrations, and validated models. Standardized workflows, population-appropriate exposure targets, prospective validation, and evidence of improved clinical outcomes are essential for broader implementation.

Pharmaceutics doi: 10.3390/pharmaceutics18091153

Authors: Ewoud Vaneeckhaute Julie Van Hooste Pasquinel Weckx Andrew Teasdale Ruud Cuyvers Jonathan Hammond Daniel Wood Ward D’Autry Laura Martin Eric Breynaert

Background/Objectives: Oral solid dosage (OSD) forms are historically categorized as low-risk vectors for leachables under the assumption that solid-state matrices impose severe kinetic barriers against migrant migration and reactivity. This study investigates an unexpected unknown impurity, detected as part of a routine ICH Q3B testing program to control drug product impurities in a blister-packed tablet. Since the impurity exceeded the ICH Q3B identification threshold applicable for this drug product (1.0% w/w relative to API or 5 µg total daily intake (TDI), whichever is lower), a forensic investigation into the origins and identity of the impurity was performed. Methods: Placebo and packaging line studies were performed to isolate the origin of the impurity. Systematic structure elucidation was carried out using high-resolution mass spectrometry (HRMS), isotopic fine structure analysis, and de novo chemical synthesis. To enable definitive characterization, preparative HPLC was utilized to isolate the compound for high-field 2D-NMR spectroscopy (801 MHz). Results: The impurity was proven to be independent of active pharmaceutical ingredient degradation, forming only when tablet excipients were stored in sealed PVC blister packaging. Co-incubation of the PVC heat stabilizer derivative dimethyltin bis(2-ethylhexyl mercaptoacetate) (DMTE) with pyrrolidin-2-one (a povidone excipient degradation product) generated an identical chromatographic and MS/MS spectral profile. NMR spectroscopy unambiguously identified the structure as 2-ethylhexyl 2-((5-oxopyrrolidin-2-yl)thio)acetate. Conclusions: This study provides the first direct evidence of an organotin-catalyzed solid-state reaction between a packaging leachable and a tablet excipient under 25 °C/60% RH storage conditions. While this implies that the impurity should not be evaluated under the ICH Q3B guidelines, the findings directly challenge the assumed “low-risk” status of OSD packaging regarding leachables and highlight the necessity for interaction-focused, chemistry-based risk assessments, as outlined in emerging ICH Q3E guidelines.

Pharmaceutics doi: 10.3390/pharmaceutics18091152

Authors: Filip Dugonik Urška Jeršič Lejla Kač Uroš Maver Tina Maver

Background: In vitro skin models are widely used in dermal and transdermal drug development, yet their pharmacokinetic interpretation often remains endpoint-dependent and does not fully capture cutaneous disposition, including partitioning, local metabolism, or clearance-like removal. This review evaluates the utility of these models through a comprehensive pharmacokinetic process framework. Methods: Using a structured literature search, we critically compared data from studies utilizing a range of in vitro skin models, including synthetic membranes, ex vivo human skin, reconstructed human epidermis, full-thickness equivalents, and advanced bioprinted or microfluidic platforms. The assessment focused on model performance across the key processes of absorption, distribution, metabolism, and elimination. Results: Our analysis reveals that alternative barriers exhibit no uniform direction of bias relative to human skin; model rank order varies with compound, formulation, and dose, so that the available evidence does not currently support a single universal scaling relationship, although calibration within a defined applicability domain may remain possible. Synthetic membranes provide reproducible platforms for formulation ranking but lack biological retention and metabolism. Ex vivo human skin remains the closest reference for permeation and mass balance, though donor variability and storage sensitivity limit standardization. Reconstructed epidermis supports standardized permeation and epidermal targeting, while full-thickness equivalents permit dermal retention analysis. Advanced perfused constructs extend experimental capabilities, but their predictive performance remains largely uncharacterized. Conclusions: No single model universally replicates human pharmacokinetics. Model selection must therefore be carefully aligned with the specific pharmacokinetic process of interest, the model’s biological capacity, and the intended context of use to ensure clinically relevant data interpretation.

Pharmaceutics doi: 10.3390/pharmaceutics18091151

Authors: Mahmoud M. Mokhtar Rahma Wael Soliman Jassy Salim Shrouk Khaled Mohamed Lobna Abdelsalam Rehab Abdelmonem

Nanoparticles have gained considerable attention as important platforms in pharmaceutical and biomedical research due to their size, morphology, surface chemistry, and colloidal behavior, which can affect drug delivery, antimicrobial activity, diagnostics, imaging, and therapeutic performance. However, conventional chemical and physical synthesis routes may require hazardous reagents, high energy demands, and procedures that raise environmental, safety, and scalability concerns. In this context, biologically mediated synthesis has emerged as an approach in which plants, microorganisms, including fungi and algae, and isolated biomolecules contribute to nanoparticle formation by acting as reducing, capping, and stabilizing agents. This narrative review evaluates these routes as potential green strategies, covering formation principles, biological sources, nanoparticle classes, characterization challenges, pharmaceutical applications, comparison with conventional synthesis, and limitations. The reviewed evidence indicates that the composition of biological sources and reaction conditions can shape nanoparticle size, morphology, surface chemistry, stability, and biological activity, highlighting green synthesis as a design-dependent process rather than a simple substitute for chemical reducing agents. Integrated characterization using optical, spectroscopic, diffraction, microscopic, and colloidal techniques is essential for interpreting nanoparticle identity and reproducibility. Biologically mediated nanoparticles show experimental and preclinical potential in drug delivery, antimicrobial therapy, anticancer research, biosensing, imaging, and diagnostics; however, most evidence remains preclinical and model-dependent. Translation remains limited by source variability, batch inconsistency, scale-up and purification challenges, stability, incomplete safety evidence, and regulatory uncertainty. Progress will require standardized process control, comparative life cycle and techno-economic assessment, scalable manufacturing, and rigorous pharmacokinetic, biodistribution, toxicological, and regulatory validation.

Pharmaceutics doi: 10.3390/pharmaceutics18091150

Authors: Miguel Ferreira Neelum Aziz Yousafzai Lital Ben Naim Bahar Ataeinia Umar Mahmood Pedram Heidari

Background: Prostate cancer (PCa) remains a significant health challenge, requiring innovative treatments. Here, we evaluated the efficacy of Talazoparib-loaded liposomes incorporating a targeting moiety, in combination with BI 2536, in PCa. This study develops lipid nanoparticles (LIPOs) targeting prostate-specific membrane antigen (PSMA) for better drug delivery. Methods: Two new PSMA-targeting agents, DUPA-DSPE and DUPA-PEG-DSPE, were created and incorporated into LIPOs via thin-film hydration. LIPOs’ size and morphology were characterized using Dynamic light scattering, and structural analysis was done via TEM imaging. In vitro cytotoxicity and cell viability were assessed using an MTT assay, while in vivo radioactivity was evaluated by PET imaging, and radioactive activity was quantified using a γ-counter. Results: These LIPOs demonstrated excellent stability and specific binding to PSMA-expressing prostate cancer cells in vitro. In vivo, mouse model studies showed that PSMA-targeted LIPOs accumulated preferentially at tumor sites. Talazoparib, a PARP inhibitor with low solubility and cytotoxicity, has limited use in PCa. Conclusions: These findings provide proof-of-concept evidence that PSMA-targeted DUPA-PEG LIPOs can facilitate Talazoparib delivery and, particularly in combination with BI 2536, may enhance antitumor activity in PCa models. Further studies with expanded control groups and larger cohorts are warranted to validate the therapeutic efficacy of this combination strategy and support its further development for PCa treatment.

Pharmaceutics doi: 10.3390/pharmaceutics18091149

Authors: Mengqi Zhao Philip Lazarus

Introduction: The use of Δ8-tetrahydrocannabinol (Δ8-THC) has become increasingly prevalent in the United States, but its metabolic pathways remain poorly characterized. This study aimed to identify the key enzymes responsible for the 11′-hydroxylation of Δ8-THC, its primary metabolic pathway, and assess the effect of metabolizing enzyme genotype on this activity. Results: The intrinsic clearance of recombinant (r) CYP2C9 was >6200−fold higher than that observed for rCYP2C19, the only other tested CYP450 with detectable activity against Δ8-THC. The Km,u of rCYP2C9 (0.013 ± 0.0032 µM) was comparable to that observed in human intestinal microsomes (0.0076 ± 0.00067 µM), but it was substantially lower than that observed for human liver microsomes (0.73 ± 0.065 µM). Inhibition of 11-OH-Δ8-THC formation in rCYP-overexpressing microsomes was observed when using CYP probe inhibitors (sulfaphenazole for CYP2C9 and tranylcypromine for CYP2C19). The intrinsic clearance was markedly reduced in CYP2C9 variant microsomes, with 20− to 60−fold decreases observed for CYP2C9*2 (59 ± 7.9 µL·min−1·mg−1 protein), CYP2C9*3 (34 ± 3.6 µL·min−1·mg−1 protein), CYP2C9*8 (96 ± 13 µL·min−1·mg−1 protein), and CYP2C9*9 (34 ± 11 µL·min−1·mg−1 protein) as compared to wild-type CYP2C9*1 (6028 ± 266 µL·min−1·mg−1 protein). Conclusions: Given that the expression of CYP2C9 is much higher than that of CYP2C19 in both human intestine and liver, these data suggest that CYP2C9 is the primary enzyme responsible for intestinal 11′-hydroxylation of Δ8-THC and that other untested CYPs may be important in hepatic Δ8-THC hydroxylation. In addition, the reduced 11-OH-Δ8-THC formation activity observed with CYP2C9 variants (*2, *3, *8, *9) suggests interindividual variability in Δ8-THC disposition.

Pharmaceutics doi: 10.3390/pharmaceutics18091148

Authors: Saima Jan Gulam Rabbani Arif Tasleem Jan Khurshid Ahmad

Plant-derived bioactives offer diverse pharmacological activities, but their development is often limited by poor aqueous solubility, instability, rapid metabolism, and low tissue exposure. This review examines delivery platforms used to address these constraints, including liposomes, solid lipid nanoparticles, nanostructured lipid carriers, phytosomes, niosomes, polymeric and protein-based carriers, extracellular vesicles, and self-assembled polyphenol systems. The platforms are compared according to payload compatibility, loading and release behavior, biological barriers, manufacturing requirements, and formulation-specific safety risks. Though carriers of natural origin offer a sustainable and often less toxic alternative, their ability to induce immunogenicity, organ accumulation, and repeated-dose toxicity require product-specific assessment. The review also discusses sustainable extraction and formulation, emphasizing that green performance depends on the entire process rather than on the feedstock alone. Preclinical studies frequently report improved stability, exposure, or therapeutic activity, whereas human evidence remains limited and is concentrated in early-phase studies of curcumin and silybin formulations and a small number of plant-derived extracellular-vesicle preparations. Translation will require standardized characterization, defined critical quality attributes, scalable Good Manufacturing Practice-compliant production, batch consistency, comparative pharmacokinetic and toxicological studies, and appropriately powered clinical trials. These considerations support rational carrier selection based on the physicochemical properties of the bioactive, intended route and target, release requirements, and strength of the available evidence.

Pharmaceutics doi: 10.3390/pharmaceutics18091147

Authors: Diell Aliu Thibault Vallet Jörg Huwyler Maxim Puchkov Lorenna Pereira de Souza Eline Freitas Medeiros Wuelton Monteiro Jacqueline de Almeida Gonçalves Sachett Fabrice Ruiz Michael Buettcher Marc Pfister Lorenz von Seidlein Joel Tarning Kevin C. Kobylinski

Background/Objectives: Scabies is a disease that disproportionately affects young children. Oral ivermectin is a mainstay of scabies therapy, yet children <15 kg are excluded due to insufficient safety data and the lack of age-appropriate formulations. To address this gap, we developed CHILD-IVITAB, a 1 mg ivermectin orodispersible tablet based on the Template Inverted Particle technology, which enables rapid disintegration and intrinsic taste masking in a mechanically robust tablet. Methods: We report the pharmaceutical characterization of CHILD-IVITAB tablets. The EPIC-15 trial (NCT06404333) was a randomized clinical trial administering CHILD-IVITAB to children weighing 5 to <15 kg with uncomplicated scabies in Brazil. Acceptability was assessed at two administrations (D0, D7) using the ClinSearch Acceptability Score Test (CAST®), wherein CHILD-IVITAB was compared to a reference dataset. Results: The 40 mg tablets had a tensile strength of 1.87 ± 0.16 MPa, and met Ph. Eur. requirements for uniformity of mass and dosage units. CHILD-IVITAB tablets disintegrated in vitro in 18.3 ± 1.1 s and released >50% of the dose within 1 min in artificial saliva. Thirteen children were enrolled in the trial. The full dose was taken in 100% of the 26 evaluations, 73% of administrations elicited a neutral or positive reaction, and no dosage form alteration, food/drink co-administration, or extra device was required. The barycenter of all evaluations fell within the positive zone of the CAST® framework. Conclusions: These results support CHILD-IVITAB as a child-friendly ivermectin formulation suitable for children 5 to <15 kg, a population currently excluded from ivermectin treatment and prevention programs.

Pharmaceutics doi: 10.3390/pharmaceutics18091146

Authors: Ana Homšek Ilić Marija Jovanović Srđan Marković Sandra Vezmar Kovačević Tamara Knežević Ivanovski Petar Svorcan Radojka Savić Katarina Vučićević

Background/Objectives: Mucosal healing (MH) is a key therapeutic target in inflammatory bowel disease (IBD) associated with sustained remission, reduced hospitalisation, and improved outcomes. Its assessment relies on endoscopic evaluation, which is invasive, costly, and not always feasible. Therapeutic drug monitoring (TDM) of infliximab (IFX) and evaluation of available biomarkers may provide non-invasive tools for predicting treatment response. This study investigated the association between IFX pharmacokinetics, routinely available blood biomarkers and MH to identify potential surrogates for predicting treatment outcomes. Methods: A monocentric, retrospective, real-world study was conducted, and pharmacokinetic analysis was performed in NONMEM version 7.5, applying a model with priors to estimate individual pharmacokinetic parameters. Endoscopic findings were collected, and patients were categorised according to achievement of MH. Univariate and multivariable logistic regression analyses were performed to evaluate the predictive value of tested variables, and receiver operating characteristic (ROC) curve analysis was performed to assess discriminative performance. Results: All evaluated variables demonstrated statistically significant associations with MH (p < 0.05). IFX clearance showed a strong inverse relationship with the probability of achieving MH. Leukocyte count exhibited particularly good discriminative ability, with an area under the ROC curve of 74.04% and an odds ratio of 0.78, highlighting its potential clinical utility. Multivariable logistic regression identified a model including IFX clearance and platelet count (PLT) as the most promising predictors of MH. Significant differences between patient subgroups indicated the association of higher PLT and modestly increased IFX clearance with a lower probability of MH. Conclusions: IFX clearance represents a promising surrogate marker for MH. TDM-derived parameters coupled with routinely available biomarkers may improve treatment response assessment. Combining pharmacokinetic and biomarker-based approaches could reduce reliance on repeated endoscopies while facilitating more effective IBD monitoring in clinical practice.

Pharmaceutics doi: 10.3390/pharmaceutics18091145

Authors: Lucas M. Favre Ezechiel Romone Pierre Cuq Nicolas Masurier Anne Aubert-Pouëssel

Background/Objectives: Hybrid microparticles (HMP) based on silylated vegetable oils have recently emerged as promising carriers for the sustained delivery of poorly water-soluble drugs, offering a controlled release without a burst effect and excellent biocompatibility. However, their limited degradability and the acidic conditions required for the sol–gel encapsulation process restrict their biomedical applicability, particularly for acid-sensitive active compounds. The aim of the present study was to develop new degradable vegetable oil-based HMP with improved drug-solubilization capacity while preserving compatibility with acid-sensitive molecules through an adapted mild sol–gel process, buffered with sodium citrate. Methods: Two new modified oils were synthesized from castor oil derivatives, including a monoglyceride (glyceryl monoricinoleate) and a polylactic acid (PLA) conjugate, to enhance both solubilization capacity and degradability. The acid-sensitive, lipophilic antitumor compound JMV5038 was used as a model drug. Modified oils were evaluated for drug-solubilization capacity, and the resulting HMPs were characterized for particle size, encapsulation efficiency, in vitro release kinetics, cytocompatibility, antiproliferative activity against melanoma cells, and in vitro enzymatic degradability. The sol–gel protocol was optimized to minimize drug degradation during encapsulation. Results: Microparticles (20–111 µm) exhibited a hybrid composition with efficient cross-linking. Encapsulation efficiencies of 85–100% were achieved, aided by up to 10-fold higher drug solubility in the new oils compared to original ICOe. Release kinetics were sustained and linear (11–53 days), fastest for IGRe- and slowest for ICOLAe-based HMP. Enzymatic degradation reached up to 46% after 30 days in the presence of fungal lipases, twofold higher than previously published ICOe microparticles, with ICOLAe degrading fastest due to PLA incorporation. Encapsulated JMV5038 retained its cytotoxic efficacy against A375 melanoma cells. Conclusions: These results highlight the promising use of such microparticles as versatile carriers for the prolonged release of lipophilic and acid-sensitive drugs.

Pharmaceutics doi: 10.3390/pharmaceutics18091144

Authors: Wei Sang Yijun Huang Jiahui Wen Weiqing Yue Ajing Wu Jie Su Ziliang Zheng Jie Li Ruiping Zhang

Background: Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer characterized by significant treatment challenges and poor prognosis. Traditional therapies such as surgical resection, radiotherapy, and chemotherapy often suffer from limitations including insufficient therapeutic specificity, potential damage to normal tissues, and limited imaging capability. The integrated diag{Pareja, 2018 #13}nosis-treatment platform combines diagnostic and therapeutic functions, offering a novel approach for real-time tumor monitoring and precision therapy. Methods: Accordingly, this study designed and developed an integrated diagnostic and therapeutic platform called A2S-TT NPs, utilizing self-assembled nanoparticles incorporating organic conjugated molecules for precise photodynamic therapy (PDT) guided by dual modalities of Near-infrared II (NIR-II) fluorescence imaging and photoacoustic imaging (PA). Results: Under 808 nm laser excitation, A2S-TT NPs efficiently generated reactive oxygen species (ROS) at the tumor site, inducing mitochondrial membrane potential disruption and DNA damage to activate apoptosis. In vitro experiments demonstrated that A2S-TT NPs exhibited intense NIR-II fluorescence and photoacoustic signals, showcasing excellent dual-modal imaging potential. Moreover, it demonstrated minimal cytotoxicity in the absence of light exposure but significantly enhanced tumor cell killing under laser irradiation, demonstrating superior biosafety and photodynamic efficacy. In vivo studies further confirmed that A2S-TT NPs effectively accumulated at tumor sites, enabling visualization-based monitoring through its mediated NIR-II and PA dual-modal imaging. Additionally, A2S-TT NPs-mediated PDT exhibited remarkable tumor-suppressive effects while avoiding significant damage to normal tissues. Discussion: This study constructs a novel PA/NIR-II dual-modal imaging-guided PDT platform based on donor–acceptor conjugated polymer A2S-TT NPs for integrated TNBC theranostics. PA provides high-resolution deep-tissue imaging, while NIR-II fluorescence offers high sensitivity and deep penetration; their complementary strengths support precise tumor localization and real-time therapeutic monitoring. Under dual-modal guidance, A2S-TT NPs enrich in tumors and generate abundant ROS upon 808 nm laser irradiation to boost PDT efficacy. In vitro and in vivo biosafety tests confirm its favorable biocompatibility with minimal systemic toxicity at therapeutic doses. Conclusions: In conclusion, A2S-TT NPs achieved precise, efficient, safe, and controllable PDT through dual-mode imaging using NIR-II and PA, providing a novel strategy with clinical translation potential for the integrated diagnosis and treatment of TNBC.

Pharmaceutics doi: 10.3390/pharmaceutics18091143

Authors: Myeong Kyun Yoo Su Jeong Kang Min Jeong Jo Jae Min Lee Moon Sup Yoon Seon Min Park Sinem Yaprak Karavana Dae Hwan Shin

Background: Ovarian cancer is frequently diagnosed at an advanced stage and remains one of the most lethal gynecologic malignancies. The development of multidrug resistance (MDR) during repeated chemotherapy is a major cause of treatment failure and is often associated with increased drug efflux mediated by transporters such as P-glycoprotein (P-gp). In this study, D-α-tocopheryl polyethylene glycol succinate (TPGS) and Soluplus® (SOL) mixed micelles, abbreviated as TS, were developed to co-deliver apatinib (APA), a VEGFR-2 inhibitor, and etoposide (ETP), a topoisomerase II inhibitor, for MDR ovarian cancer models. The formulation was designed to improve the aqueous dispersion, cellular accumulation, and antitumor activity of APA and ETP. Methods: APA/ETP-loaded TS micelles (APA/ETP-mTS) were prepared and characterized in terms of particle size, polydispersity index (PDI), zeta potential, encapsulation efficiency (EE), storage stability, and in vitro drug release. Anticancer efficacy was assessed using MTT assays, cellular uptake assays, and 3D tumor spheroid studies employing HeyA8-MDR cells, followed by in vivo toxicity and antitumor efficacy studies. Micellar formulations were denoted using the cargo–carrier format, where APA/ETP indicates co-loaded APA and ETP, C6 indicates coumarin-6 (C6) used as a fluorescent probe, mTS indicates TPGS/SOL mixed micelles, and mSOL indicates SOL-only micelles. Results: The selected APA/ETP-mTS formulation showed a particle size of 20.0 ± 5.1 nm, a PDI of 0.16 ± 0.05, near-neutral zeta potential, and encapsulation efficiencies exceeding 60% for both drugs. The micelles maintained colloidal stability at 4 °C for 4 weeks, although partial decreases in encapsulation efficiency were observed. Compared with APA/ETP solution, APA/ETP-mTS delayed the release of both drugs. C6-mTS showed higher intracellular fluorescence intensity than C6-mSOL, suggesting enhanced cellular accumulation associated with TPGS incorporation. APA/ETP-mTS showed greater cytotoxicity than free drugs in HeyA8-MDR monolayer cells and produced the strongest spheroid growth inhibition among the tested micellar formulations. In the HeyA8-MDR xenograft model, APA/ETP-mTS suppressed tumor growth and resulted in the lowest final tumor weight without apparent overt toxicity based on body weight and survival observations. Conclusions: These results suggest that APA/ETP-mTS is a promising micellar co-delivery platform for hydrophobic anticancer drugs in MDR ovarian cancer.

Pharmaceutics doi: 10.3390/pharmaceutics18091142

Authors: Fang Wang Xiang Deng Yuwen Zhu Xinyu Zong Yazhong Ma Hailong Yuan

Objectives: Herpetrione (HPE) is a bioactive lignan recognized for its hepatoprotective properties; however, it exhibits limited oral bioavailability. This study aimed to classify HPE within the framework of the biopharmaceutics classification system (BCS) and to develop a nanoparticle formulation. Methods: To this end, a comprehensive investigation was conducted, encompassing computer prediction analysis, equilibrium solubility measurements across the gastrointestinal pH range, Caco-2 bidirectional transportation, in situ single-pass intestinal perfusion (SPIP), and molecular docking with P-glycoprotein (P-gp). Results: In silico analyses suggested that HPE possesses low solubility and low permeability characteristics. Experimental assays revealed pH-dependent solubility and inherently low aqueous dissolution. Unlike the computer prediction results, Caco-2 studies revealed moderate permeability but a high efflux ratio, suggestive of possible P-gp substrate activity for HPE, a finding further supported by molecular docking simulations. Conversely, SPIP studies demonstrated that the effective permeability (Peff) of jejunal intestinal segments exceeded the high-permeability threshold, thereby classifying HPE as a high-permeability drug. Based on these findings, HPE was classified as a BCS class II compound. To overcome its solubility-limited absorption, a nanoparticle was developed, resulting in a marked enhancement of both solubility and dissolution rates. Conclusions: These findings underscore the importance of integrating experimental biopharmaceutical evaluations with computational tools when designing delivery systems for natural products.

Pharmaceutics doi: 10.3390/pharmaceutics18091141

Authors: Evan Gallagher Joseph W. Downey Chi-Hyeon Yoo Hongping Deng Jessie Fanglu Fu Qikai Qin Irena Bass Hsiao-Ying Wey Jacob M. Hooker Christin Y. Sander So Jeong Lee

Background/Objectives: Quantitative PET imaging of μ-opioid receptor (MOR) availability and pharmacologic engagement can advance in vivo studies of opioid receptor biology. We previously evaluated a series of 18F-labeled carfentanil analogs and prioritized the regioisomer [18F]fluorocarfentanil ([18F]FCFN) based on favorable brain kinetics, regional distribution, and naloxone-sensitive binding. Here, we provide proof-of-concept evaluation of [18F]FCFN in rhesus macaques to assess its potential for quantitative, pharmacologically responsive MOR PET. Methods: [18F]FCFN was produced by automated radiosynthesis. Dynamic [18F]FCFN PET scans were performed in two rhesus macaques under baseline conditions, after low- (0.005 mg/kg) and high-dose (0.2 mg/kg) naloxone pretreatment, and during within-scan intravenous or intranasal naloxone challenge. For arterial input scans, two-tissue compartment modeling with metabolite-corrected arterial input functions was used to estimate total distribution volume (VT). Nondisplaceable binding potential (BPND) was estimated using multilinear reference tissue model 2 with the cerebellum as the candidate reference region. Results: Automated radiosynthesis produced [18F]FCFN in high radiochemical yield and molar activity, enabling low-mass PET injections (0.09–0.32 μg). [18F]FCFN showed rapid brain uptake and MOR-consistent regional distribution, with lowest uptake in the cerebellum. Regional BPND correlated positively with VT at baseline (R2 = 0.983, p < 0.0001) and after high-dose blockade (R2 = 0.522, p < 0.004). Naloxone pretreatment reduced regional BPND in a dose-dependent manner. Within-scan intravenous and intranasal naloxone challenges also produced marked reductions in BPND. Conclusions: [18F]FCFN enables quantitative, pharmacologically sensitive MOR PET imaging in nonhuman primates at low injected masses. Concordant plasma-input and reference-tissue measures, dose-dependent blockade, and proof-of-concept within-scan displacement support its use for measuring MOR availability and dynamic antagonist engagement across administration routes.

Pharmaceutics doi: 10.3390/pharmaceutics18091140

Authors: Manru Zhang Bohan Chen Tiezheng Li Xiaolong Li Jingxian Sun Qiushuo Li Mingji Jin Zhonggao Gao Liqing Chen Wei Huang

Central nervous system (CNS) inflammation and brain tumor treatment are constrained by the heterogeneity of the blood–brain barrier (BBB) and blood–brain tumor barrier (BBTB), as well as by the sequential barriers to drug delivery across lesions, target cells and subcellular organelles. Simply increasing brain exposure does not ensure that drugs reach their actual sites of action. This review systematically examines the pathological roles and therapeutic rationales of mitochondrial, lysosomal, nuclear, endoplasmic reticulum and Golgi apparatus dysfunction in neuroinflammation and glioblastoma within a two-stage delivery framework encompassing barrier crossing, lesion accumulation, cellular uptake and subcellular organelle localization. It also summarizes key design considerations for liposomes, polymeric nanoparticles, biomimetic membrane-based carriers, exosome-like carriers and focused ultrasound-assisted delivery strategies. Furthermore, translational bottlenecks are discussed, including BBB/BBTB heterogeneity, endosomal/lysosomal escape, organelle off-targeting, long-term safety and the extrapolation of preclinical models. Finally, personalized delivery designs guided by cascade targeting, dynamic visualization-based validation and disease stratification are proposed to support precise treatment of CNS inflammation and brain tumors.

Pharmaceutics doi: 10.3390/pharmaceutics18091139

Authors: Gabriela E. Galarza-Arévalo Myriam P. Gonzalez Tania Valdiviezo-Abarca Mateo A. Salazar Alexis Debut Miryan R. Rivera María P. Romero

Background: Chronic ulcers are a major healthcare challenge due to their prolonged course, recurrence, persistent microbial colonization, and impaired tissue repair. This study developed sodium alginate–collagen hydrogel dressings reinforced with cotton fabric and functionalized with green-synthesized copper oxide nanoparticles (CuO NPs) to combine exudate absorption with localized antibacterial activity. Methods: CuO NPs were synthesized using gallic acid and orange peel extract and evaluated for cytotoxicity and antibacterial activity against Staphylococcus aureus and Escherichia coli. Sodium alginate–collagen dressings containing different glycerin concentrations were assessed for swelling capacity, morphology, structural stability, and tensile performance. The formulation showing the best overall physicochemical and mechanical performance was selected for CuO NP functionalization and antibacterial evaluation by agar diffusion. Results: The selected 50GS-NPs-CuO formulation showed an IC50 of 79.9 µg/mL. At 70 µg/mL, CuO NPs produced significant time-dependent reductions in bacterial viability, reaching 96.6% for S. aureus and 91.44% for E. coli after 72 h. The dressing containing 2% sodium alginate, 0.5% collagen, and 2.5% glycerin exhibited the highest swelling capacity (476.82 ± 3.80%) and tensile strength (148.7 ± 5.5 N). CuO-functionalized dressings showed concentration-dependent antibacterial activity, with consistently greater inhibition against S. aureus than E. coli. Conclusions: The developed dressing integrates high fluid absorption, mechanical stability, and localized antibacterial activity, providing a promising platform for advanced chronic-wound management.

Pharmaceutics doi: 10.3390/pharmaceutics18091138

Authors: Dai Shi Wujian Mao Yutao Xie Pan Zhou Minqiang Hu Yuxia Liu Dengfeng Cheng

Malignant tumors are the second major disease threatening human health, and their early diagnosis can significantly improve patients’ prognosis. [18F]FDG PET/CT is currently the most important method in nuclear medicine for tumor diagnosis and staging. However, due to variations in the glucose uptake characteristics of tumors, some tumors with low FDG uptake (e.g., gastric mucinous adenocarcinoma and indolent lymphoma) are prone to false-negative results. Therefore, new tumor-specific imaging tracers are urgently needed. Due to the significant differences in target expression among various tumor cells, the method of focusing on a single target is unlikely to work for all tumors. Therefore, researchers are exploring the possibility of targeting tumor-associated macrophages (TAMs) in the tumor microenvironment for imaging because this offers several advantages. First, unlike most tumors, which originate from epithelial cells, TAMs arise from monocyte–macrophage polarization, leading to greater target specificity on their surfaces. Second, TAMs are the most abundant non-tumor cells in the tumor microenvironment, ensuring a certain level of expression abundance. Furthermore, numerous preclinical studies and clinical trials targeting TAMs with immunotherapy are currently underway, making the non-invasive assessment of certain TAM targets’ expression levels a critical issue that needs to be addressed. This article summarizes the potential targets and nuclear medicine tracers for targeting TAMs.

Pharmaceutics doi: 10.3390/pharmaceutics18091137

Authors: Tomona Yukimura Hirono Ito Takahiro Suzuki Toshinobu Seki Tomohiro Seki

Background: Although intracellular delivery via endocytosis is a promising strategy for drugs acting in the cytosol or nucleus, many macromolecular therapeutics remain trapped within endosomal/lysosomal compartments, limiting efficient cytosolic delivery. In this study, we designed a novel functional hyaluronic acid (HA)-based polymer, HA–octahistidine (His8), by conjugating His8 to CD44-targeting HA, and evaluated its physicochemical properties, cellular uptake, and endosomal escape capability. Methods: HA–His8 was synthesized via 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide (EDC/NHS)-mediated coupling and characterized using 1H NMR spectroscopy. Particle size and zeta potential were measured via dynamic light scattering, and buffering capacity was evaluated using acid–base titration. Cellular uptake and intracellular localization were investigated in CD44-high MDA-MB-231 and CD44-low MCF-7 cells via confocal laser scanning microscopy. Cytotoxicity was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results: Successful conjugation of His8 was confirmed with a degree of substitution of 4.66 mol% relative to HA carboxyl groups. HA–His8 exhibited a nanoscale hydrodynamic diameter under physiological conditions, which increased under acidic conditions together with changes in zeta potential. HA–His8 exhibited higher buffering capacity than free His8 and was preferentially internalized by CD44-high MDA-MB-231 cells compared with MCF-7 cells. Compared with unmodified HA, HA–His8 exhibited lower colocalization with LysoTracker that decreased over time, indicating reduced retention within acidic vesicles. HA–His8 also exhibited low cytotoxicity over the tested concentration range. Conclusions: His8 modification alters HA intracellular localization while preserving CD44 targeting, thereby facilitating endosomal escape. These findings highlight HA–His8 as a potential platform for the cytosolic delivery of macromolecular therapeutics, including proteins, peptides, and nucleic acids.

Pharmaceutics doi: 10.3390/pharmaceutics18091136

Authors: Payal Ganguly Elena A. Jones Peter V. Giannoudis Giuseppe Tronci

Antimicrobial resistance (AMR) presents a global health challenge that is projected to cause nearly 2 million deaths annually by 2050. With a growing ageing population and the rise of antibiotic-resistant strains globally, developing effective alternatives to combat infections while reducing AMR risks is critical. Over recent decades, antibiotic-free strategies have emerged as promising approaches, offering advantages over traditional antibiotics and reducing the likelihood of resistance. This review highlights antibiotic-free antimicrobial strategies for applications in chronic wound care, hard tissue repair, personal protective equipment, as well as hospital hygiene and infection control. We begin by outlining the broader AMR problem and then examine natural strategies—including the use of agents such as manuka honey—followed by synthetic approaches involving nanomaterials and metal–organic frameworks (MOFs), as well as biological and bioinspired antimicrobial strategies, involving antimicrobial peptides, antibodies, bacteriophages, extracellular vesicles, and macrocycles. Finally, we discuss innovations in artificial intelligence (AI)-assisted antimicrobial development and stimulus-responsive materials, before introducing a conceptual design framework for antibiotic-free antimicrobial materials. This review provides a comprehensive assessment of emerging antibiotic-free strategies to guide the development of next-generation antimicrobial solutions that reduce reliance on antibiotics and mitigate AMR in clinical and healthcare settings.

Pharmaceutics doi: 10.3390/pharmaceutics18091135

Authors: Louisa Bourel Sixtine Gilliot Zeina Boulos Yousra Alouadni Kévin Le Duc Mohamed Riadh Boukhris Laurent Storme Anthony Martin Mena Pascal Odou

Background/Objectives: Etilefrine is a sympathomimetic agent increasingly used off-label as a rescue therapy for refractory neonatal chylothorax. For continuous intravenous administration in neonates, the commercially available 10 mg/mL formulation requires substantial dilution to a concentration of 4 µg/mL. However, stability data for these highly diluted preparations are currently lacking. Hence, this study aimed to assess the short-term physicochemical stability of etilefrine diluted to 4 µg/mL in 0.9% sodium chloride or 5% dextrose. Methods: Etilefrine was quantified using a high-performance liquid chromatography method coupled with ultraviolet detection. Diluted solutions were prepared in 10 mL polypropylene syringes, maintained at room temperature, and analyzed daily for 96 h. Physicochemical stability assessment included visual inspection for physical changes, assays of drug concentration, pH, osmolality, and subvisible particle counts. Results: Throughout the 96 h, no visible changes, such as precipitation, discoloration, or haze, were observed. Etilefrine concentrations in both isotonic diluents remained within 10% of the initial value. There were no significant variations in pH and osmolality. Subvisible particle counts remained below the limits established by the European Pharmacopoeia. Conclusions: Etilefrine diluted to 4 µg/mL in 0.9% sodium chloride or 5% dextrose in syringes remained physicochemically stable for 96 h at room temperature. These findings provide practical stability data to support the preparation and administration of diluted etilefrine, and may facilitate the standardization of its use in future clinical studies of refractory neonatal chylothorax.

Pharmaceutics doi: 10.3390/pharmaceutics18091134

Authors: Xiaoya Li Lingling Li Yongfang Yuan Changxin Sun Yajie Wang Xuefei Wang Yixuan Feng Min Wu Longtao Liu

Background: Polydatin has shown anti-inflammatory, antioxidant, and cytoprotective effects in preclinical studies. Its development is limited by low oral bioavailability, extensive metabolism, and poorly defined tissue exposure. Objectives: This review summarizes the pharmacology and delivery systems of polydatin, distinguishes nano from non-nano formulations, and examines the evidence for improved exposure, tissue delivery, safety, and clinical translation. Methods: Relevant studies were identified through a structured PubMed search and assessed by formulation type, administration route, comparator, and reported endpoints. Results: Delivery strategies include liposomes, polymeric and polysaccharide nanoparticles, lipid-based formulations, inclusion complexes, solid-state systems, hydrogels, local matrices, and targeted systems. Only a few oral formulations have been compared directly with free polydatin using quantitative pharmacokinetic measurements. Many injectable and local studies report release, cellular uptake, fluorescence distribution, or pharmacodynamic effects, but these findings do not by themselves demonstrate improved bioavailability or drug-specific targeting. Evidence on repeat-dose toxicity, immunogenicity, carrier clearance, and manufacturing consistency remains limited. No clinical study of a polydatin nanocarrier was identified. Conclusions: Further development requires appropriate free-polydatin controls, measurement of polydatin and its metabolites, exposure–response analysis, fuller safety assessment, and reproducible product characterization.

Pharmaceutics doi: 10.3390/pharmaceutics18091133

Authors: Carla González de la Cruz Levin Thomas Carmen Mata-Martín Juan Antonio Villatoro-García Fernando de Andrés Idian González-Rodríguez Idilio González-Martínez Eva M. Peñas-Lledó Adrián LLerena

Background: Interindividual variability in sertraline pharmacokinetics remains substantial, but the relative contributions of pharmacogenetic (PGx), demographic, clinical, and prescribing determinants have not been comprehensively evaluated in routine clinical practice. The study aimed to evaluate PGx, demographic, clinical, and prescribing determinants of sertraline plasma concentrations, sertraline plasma concentration-to-dose ratio (C/D), and non-dose-normalized sertraline-to-norsertraline metabolic ratio (MR), with log10-transformed dose-normalized sertraline/norsertraline metabolic ratio (LogMRDN) and log10-transformed MR (LogMR) evaluated as exploratory derived pharmacokinetic outcomes in real-world clinical settings. Methods: This prospective real-world PGx study included 153 patients receiving sertraline therapy who participated in the MedeA clinical PGx implementation program. Associations of genotype-predicted phenotypes for CYP2C19, CYP2B6, CYP2D6, CYP2C9, and CYP3A4, CYP2C:TG haplotype status, demographic, clinical, and prescribing characteristics with sertraline concentration, C/D, MR, and LogMRDN were evaluated using univariable analyses. Separate multivariable linear regression models were fitted for these outcomes and LogMR using 140 complete cases. Results: CYP2C19 gPM status was associated with higher sertraline concentrations (β = 72.10, 95% CI: 21.36–122.84; p = 0.01). Although a higher sertraline C/D ratio was observed in the conventional multivariable model with CYP2C19 gPM (β = 1.46, 95% CI: 0.85–2.08; p < 0.001), this association was not retained after HC3 heteroscedasticity-robust standard errors were applied (β = 1.46, 95% CI: −0.09–3.02; p = 0.07). In a small exploratory subgroup analysis, patients with combined CYP2C19 and CYP2B6 poor-metabolizer status (n = 2) had sertraline concentrations exceeding 150 ng/mL. Conclusions: The study findings provide further insights into the relevance of CYP2C19 in sertraline pharmacokinetics.

Pharmaceutics doi: 10.3390/pharmaceutics18091132

Authors: Zofia Śledzikowska Karolina Żylińska Natalia Makaruk Dominika Kubicka Filip Adam Błotniak Napoleon Waszkiewicz

Oral nanocarrier development for inflammatory bowel disease (IBD) is complicated by inflammation-dependent changes in mucus, epithelial integrity, and immune-cell interactions. This critical narrative review examines comparative evidence from in vitro systems, animal models, and human intestinal tissue, alongside clinical studies, to assess how barrier state modifies formulation performance. Across the reviewed studies, particle size alone does not consistently predict lesion accumulation, while surface chemistry, coating, geometry, and stability in gastrointestinal fluids influence mucus interaction, cellular access, and payload release. Tissue-associated fluorescence, increased permeability, and cellular uptake do not independently establish intact-carrier transport or productive delivery. Clinical trials of selected oral nano-enabled formulations report improvements in some disease outcomes but do not establish inflammation-selective delivery as the underlying mechanism. We propose an evidence-informed benchmarking framework linking characterization in biological media to mucus transport or retention, epithelial–immune responses, spatial localization, target engagement, and safety. Matched healthy and inflamed comparators are central to testing selectivity, and preclinical findings must be distinguished from clinical validation. Artificial intelligence may support barrier-state-aware formulation optimization but requires standardized datasets and prospective experimental validation. Given the heterogeneous evidence base and limited direct human delivery studies, prospective validation should prioritize local payload delivery and therapeutic benefit without exacerbating intestinal barrier injury.

Pharmaceutics doi: 10.3390/pharmaceutics18091131

Authors: Kulzhan Berikkhanova Yernur Zakirov Askhat Zhilkaidarov Nurgul Daniyeva Erlan Taigulov Saken Kozhakhmetov Ardak Omarbekov Beibarys Amankeldin Gulyash Tanysheva Nurzhan Berikkhanov Yessenkhan Sultan Zhanas Baimagambet Isah Inuwa Saule Aliyeva

Background/Objectives: Diabetic foot ulcers are characterized by persistent inflammation, impaired angiogenesis, and delayed tissue regeneration. Epidermal growth factor (EGF) is an important regulator of wound repair; however, its therapeutic application may be limited by rapid degradation and poor retention within the wound environment. This study investigated the therapeutic efficacy of local administration of EGF and ceftriaxone using autologous red blood cell-derived carriers (RBCDCs) in an experimental model of diabetic wound healing in rats. Methods: Experiments were conducted on 30 albino Wistar rats (250 ± 20 g) with experimentally induced diabetic wounds. In the RBCDC treatment groups, separately prepared drug-loaded RBCDC formulations containing EGF or ceftriaxone were locally administered into the wound. The corresponding comparison groups received the same active agents in free form according to an identical treatment schedule, while an additional control group received topical Levomekol. Results: RBCDC-mediated administration of EGF and ceftriaxone significantly accelerated wound closure and was associated with earlier resolution of inflammation, granulation tissue maturation, re-epithelialization, collagen deposition, and angiogenesis compared with the corresponding free-drug formulations. On day 9, tissue EGF concentration was higher in the RBCDC (EGF + Ctx) group than in the corresponding free-drug group. Additional formulation characterization demonstrated ceftriaxone encapsulation and in vitro release from RBCDCs, as well as in vitro EGF release from EGF-loaded RBCDCs. Conclusions: Autologous RBCDCs represent a promising local drug-carrier platform for the administration of regenerative and antimicrobial agents in experimental diabetic wounds. Further studies are required to establish local pharmacokinetics, systemic exposure, carrier stability, and the mechanisms responsible for the observed therapeutic effects.

Pharmaceutics doi: 10.3390/pharmaceutics18091130

Authors: Juan Carlos Ruiz Ramirez Adrián Gómiz Sáez María Encarnación Martínez Madrid Alice Charlotte Viney José María Alonso Herreros Pilar Almela Rojo

Background/Objectives: Oseltamivir is a widely used antiviral agent indicated for the treatment of influenza and plays a central role in pandemic preparedness strategies. Although an extemporaneously prepared 15 mg/mL oral solution is recognized in formularies, preparing large volumes during a pandemic requires extended storage options. However, no stability studies are currently available for an oseltamivir 15 mg/mL oral solution stored under freezing and subsequent refrigerated conditions for the duration of a standard treatment. The aim of this study was to evaluate the physicochemical and microbiological stability of a 15 mg/mL oseltamivir oral solution prepared from the active pharmaceutical ingredient (API) and packaged in amber glass containers. Methods: The oral solution was formulated using oseltamivir phosphate API, sodium benzoate as a preservative, and purified water and then packaged in 125 mL Type II amber glass bottles, allowing for thermal expansion. The samples were stored at −20 ± 2 °C for up to 75 days, followed by refrigerated storage (5 ± 3 °C) after thawing for up to 10 days. Chemical stability was assessed using a validated HPLC method in accordance with ICH guidelines and was defined as 90–110% recovery of the initial concentration. Physical stability (color, pH, particulate matter, crystallization, and homogeneity) and microbiological stability were also evaluated. Results: The HPLC method demonstrated excellent linearity, precision, and accuracy. Oseltamivir concentrations remained within the predefined acceptance limits throughout the 75-day study period under freezing conditions, with no significant changes in pH, color, or particulate formation. After thawing, the drug concentration continued to remain fully stable and within the required limits for up to 10 days under refrigerated conditions. Despite the prolonged storage and phase changes, no significant changes in physical parameters were observed, and microbiological testing confirmed the absence of aerobic, anaerobic, and fungal microorganisms on the final day of the study. Conclusions: Oseltamivir 15 mg/mL oral solution in amber glass bottles is physicochemically and microbiologically stable for up to 85 days (75 days under frozen conditions plus 10 days under refrigeration after thawing).

Pharmaceutics doi: 10.3390/pharmaceutics18091129

Authors: Gergely Molnár Balázs Simon Arash Mirzahosseini Máté Dobó Gergely Dombi Béla Fiser Ali Mhammad Vivien Bárdos Rita Szolláth Zoltán-István Szabó András Marton Kamilla Varga Tamás Tábi Imre Boldizsár Andrea Horváth Orsolya Dobay Szilvia Bősze György Tibor Balogh Gergő Tóth

Background/Objectives: Stereochemistry can substantially influence the biological activity and pharmaceutical properties of small molecules. This study investigated the stereochemistry-dependent biological activity and chiral recognition of novel naphthylethyl thiourea enantiomers using an integrated biological, chromatographic, and computational approach. Methods: Ten pairs of naphthylethyl thiourea enantiomers were synthesized from enantiomerically pure precursors and characterized. Antiproliferative and antibacterial activity, chiral HPLC behavior on polysaccharide- and protein-based stationary phases, molecular docking, and multivariate relationships were evaluated. Results: Target-dependent enantioselectivity and substituent-dependent activity patterns were observed. All enantiomeric pairs were chromatographically distinguished under at least one condition, and selected compounds showed selector- and mobile-phase-dependent changes in elution order. AGP-based chromatography showed stereoselective recognition for a subset of compounds, while computational analyses provided complementary support for the experimental trends. Conclusions: Stereochemistry and substitution jointly influence biological and chromatographic behavior in this thiourea series. Integrated biological, chromatographic, and computational profiling provides a useful framework for early enantiomer-resolved developability assessment.

Pharmaceutics doi: 10.3390/pharmaceutics18091128

Authors: César Alas-Pineda Dennis J. Pavón-Varela Kristhel Gaitán-Zambrano Carlos Coto-Tejeda Jhacely Medina-Mejía Nelly Andrews Interiano Gustavo Ferrer

Intranasal drug delivery is noninvasive, rapid-acting, and well suited to self-administration, yet the real-world performance of nasal sprays depends on the interaction among formulation, device mechanics, nasal anatomy, and user technique rather than on the active ingredient alone. This narrative review examines how aerosol science, regional targeting, and human factors jointly shape intranasal performance, using GentleMist Technology™ as a representative example of emerging device-centered development. Computational fluid dynamics (CFD) and anatomically informed in vitro models indicate that device geometry, plume characteristics, and administration angle can significantly alter where a dose is deposited, while human factors research identifies priming, positioning, and angulation as key determinants of successful over-the-counter (OTC) use. We summarize early feasibility data suggesting that angle-optimized administration may increase posterior nasopharyngeal delivery and that essential tasks are feasible for lay users, with priming emerging as a specific usability vulnerability. We also review preliminary clinical evidence on a chlorpheniramine maleate (CPM)-based intranasal formulation in allergic rhinitis and viral upper respiratory illness. Although the strongest device-specific findings remain preliminary and require independent replication, they support an emerging shift from a formulation-centered model toward an integrated device–formulation–user paradigm.

Pharmaceutics doi: 10.3390/pharmaceutics18091127

Authors: Özge Esen Yigit Alf Lamprecht

Background/Objectives: Transdermal delivery of hydrophilic drugs remains limited by poor partitioning into the lipid-rich stratum corneum (SC). This study systematically investigated how ternary nanoemulsion composition and surfactant architecture jointly influence the transdermal delivery of salbutamol and whether the resulting composition–performance relationships are preserved across different skin models. Methods: Salbutamol-loaded nanoemulsions were prepared by the phase inversion temperature (PIT) method across a predefined ternary design space using two non-ionic surfactant systems: polyoxyl castor oil and polyoxyl hydroxystearate. Physicochemical characterization, ternary compositional mapping, in vitro permeation testing, generalized additive modeling (GAM), and attenuated total reflectance–Fourier transform infrared (ATR-FTIR) spectroscopy were combined to evaluate formulation-dependent transport across pig and mouse skin models, complemented by exploratory human-skin experiments. Results: Among the nanoemulsion formulations, pig skin showed the highest salbutamol permeation, with flux values reaching approximately 390 µg/cm2·h. Within the PHS-based system, mouse skin showed lower permeation and a stronger dependence on formulation composition than pig skin, while the simple aqueous vehicle also produced comparatively low permeation in the murine model. The aqueous-vehicle control produced substantially higher permeation than the nanoemulsions in pig skin but lower permeation in mouse skin, while receptor-phase salbutamol concentrations remained below the limit of quantification in human skin. Across both surfactant systems, the most favorable nanoemulsion-mediated permeation was generally associated with water-rich formulations containing comparatively low surfactant levels, whereas highly surfactant-rich regions showed reduced flux despite marked lipid- or protein-associated spectral changes in the descriptive ATR-FTIR analysis. Regression analyses suggested that droplet size and viscosity alone could not consistently explain permeation behavior, whereas compositional modeling revealed pronounced non-linear effects of the water–surfactant–oil balance. Conclusions: Overall, this study demonstrates that nanoemulsion-mediated delivery of hydrophilic drugs is governed primarily by ternary composition, with formulation effects varying across skin models. These findings highlight the importance of composition-based formulation design and appropriate skin-model selection during the development of transdermal systems for hydrophilic drugs.

Pharmaceutics doi: 10.3390/pharmaceutics18091126

Authors: Na Zhao Xiaoqian Zhang Jing Luo Xiaoyue Zhang Yonghong Zhao Jiang Liu Le Li Chenglin Hong Shiguo Sun

Background/Objectives: Cervical cancer cells evade chemotherapy by upregulating antioxidant defenses (e.g., glutathione (GSH)). A platform that simultaneously amplifies ROS and suppresses antioxidant defense is a low toxicity strategy. This study presents a local nanoplatform that combines chemodynamic therapy (CDT) with chemotherapy for the localized treatment of cervical cancer. Methods: Diallyl trisulfide (DATS) was loaded into the echinacoside (ECH)-copper metal–phenolic networks (MPNs) nanoparticles (ECD NPs) through a one-step coordination assembly method. The nanoparticles were incorporated into a poloxamer/HPMC thermosensitive hydrogel for vaginal delivery. The formulation was characterized for size, drug loading, sol–gel transition, and pH-responsive release. Antitumor activity was evaluated in SiHa and HeLa cells via Cu2+ uptake, GSH depletion, ROS accumulation, apoptosis markers, and viability. In vivo efficacy and biosafety were assessed in an orthotopic cervical cancer model. Results: ECD NPs showed uniform size (~135 nm), high DATS loading (~27.6%). The ECD NPs-loaded hydrogel exhibited a sol–gel transition at 36.7 °C. The ECD NPs-loaded hydrogel released 16.4% of DATS at pH 7.4, 32.4% at pH 4.5, 64.8% at pH 6.5, and 80.9% at pH 5.6 over 24 h. Release was minimal at vaginal pH, clearly triggered at tumor pH, and fastest at lysosomal pH, confirming pH-responsive behavior. The ECD NPs-loaded hydrogel enhanced Cu2+ uptake, depleted GSH, elevated ROS, and reduced cell viability to <50% at 80 μg/mL. In the orthotopic model, the ECD NPs hydrogel achieved a tumor inhibition rate of 87.81%, significantly outperforming free DATS (65.3%) and blank MPN (55.62%) hydrogels, with no evident systemic toxicity. Conclusions: The ECD NPs hydrogel triggers a Cu2+-driven ROS/GSH cascade that combines copper-mediated oxidative stress with DATS-induced apoptosis for enhanced antitumor activity. Its vaginal localization, pH-responsive release, and biosafety profile support further evaluation for cervical cancer therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091125

Authors: Mona M. Mostafa Shaimaa Mosallam Mai M. Eltaweel Maha M. Amin Jawaher Abdullah Alamoudi Heba Mohammed Refat M. Selim Mira Magdy William Shady M. Abd El-Halim

Background/Objectives: Repurposing existing drugs may represent a promising strategy for effective cancer therapy. This study was the first to investigate the augmented antitumor therapeutic effect achieved by co-incorporating the NSAID Niflumic acid (NIF) with ceramides into PEGylated cerosomes (NIF-loaded PEG-CERs) in a novel platform that targets specifically the MAPK-ERK signaling pathway and miR-21-5p modulation. Methods: The prepared formulae were statistically optimized utilizing a full factorial design and the optimal formula (C5) was further incorporated into a topical gel and evaluated for ex vivo rat skin permeation, and tested in vivo in a subcutaneous solid Ehrlich carcinoma (SEC) mice model. Results: The optimal formula (C5) showed tubular elongated morphology with higher EE% (96.71 ± 0.0), lower vesicular size (VS) and PDI values, 292.95 ± 0.78 nm and 0.47 ± 0.0 respectively. A high ZP value (−37.5 ± 0.57 mV) was in accordance with stability results showing good stability of the optimal formula (C5). Permeability studies exhibited 2.02-fold higher skin permeation compared to pure NIF gel. A significant decrease in tumor volume and marked improvement in survival rate in SEC mice were confirmed by downregulation of EGFR, ERK1, ERK2, and miR-21-5p expression. Furthermore, an increase in total antioxidant capacity and caspase-3 levels was observed, accompanied by significant suppression in cyclin D1, MMP-2, COX-2, and MDA levels. Finally, histopathological analysis revealed the superior antitumor effect of C5 gel together with immunohistochemical assay showing the lowest BCL-2-positive staining, indicating the restoration of physiological apoptotic balance. Conclusions: Based on the previous findings, NIF-loaded PEG-CERs offer augmented therapeutic potential for efficient topical skin cancer management in an SEC mice model.

Pharmaceutics doi: 10.3390/pharmaceutics18091124

Authors: Lingxin Zeng Xuan Chen Ying Li Wei Xiong

Klotho is a longevity-associated and tissue-protective protein involved in mineral metabolism, oxidative stress, inflammation, fibrosis, cellular senescence, and neurovascular homeostasis. However, nearly three decades after its discovery, no Klotho-based therapy has been approved, highlighting a translational gap that extends beyond biological validation. This review reframes Klotho translation as a pharmaceutical sciences challenge, focusing on how to convert Klotho into a druggable, manufacturable, deliverable, and clinically controllable therapeutic product. We summarize the isoform-specific properties of membrane-bound α-Klotho, soluble α-Klotho, and β-Klotho that are relevant to product design, and review the current clinical and preclinical landscape dominated by gene-, mRNA-, and antibody-based approaches. We further distinguish confirmed developability barriers, including renal handling and limited systemic persistence, from plausible risks common to macromolecular biologics, such as aggregation, chemical degradation, immunogenicity, and poor tissue penetration. Finally, we evaluate emerging delivery and formulation strategies, including viral and non-viral gene delivery, extracellular vesicles, hydrogels, ultrasound-targeted microbubbles, osmotic pumps, and long-acting protein engineering. An integrated roadmap combining molecular engineering, disease-specific delivery, pharmacokinetic/pharmacodynamic biomarkers, manufacturability assessment, and repeated-dose safety evaluation may help transform Klotho from a promising anti-aging molecule into a clinically viable biologic platform.

Pharmaceutics doi: 10.3390/pharmaceutics18091123

Authors: Raiane V. Cardoso Patricia R. Pereira Cyntia S. Freitas Yuri P. Souza Dário E. Kalume Giovani Carlo Verissimo da Costa Carlos A. Conte-Junior Vania Margaret Flosi Paschoalin

Background/Objectives: Tarin exhibits immunomodulatory and antiproliferative properties against several tumor cell lines. Nano-encapsulation in liposomes enhances its therapeutic potential by improving protein stability, bioavailability, and sustained release. Previous studies demonstrated that nano-encapsulated tarin induces cell cycle arrest, migration inhibition, apoptosis, and autophagy in triple-negative breast cancer cells; however, the molecular mechanisms underlying these effects remain poorly understood. To investigate the proteomic response elicited by nano-encapsulated tarin, MDA-MB-231 cells were treated for 24 and 48 h. Methods: Intracellular proteins were extracted, digested with trypsin, and analyzed by label-free LC-2D-MS/MS using HDMSE acquisition. Differentially expressed proteins were identified and quantified using the Progenesis QI platform, and then functional classification and pathway enrichment analyses were performed. Results: A total of 2818 proteins were identified, of which 2150 displayed time-dependent modulation following treatment. After 24 h, cells exhibited an adaptive stress response profile characterized by increased DNA repair proteins (CHEK1, CDK12), migration/remodeling factors (LAMA4, CTTN, A2M), and immune/cell cycle regulators (PER2, HLA-B), while antioxidant proteins (SOD1, GPX1) and BRCA1 were reduced, indicating oxidative stress and DNA damage. After 48 h, the proteomic profile shifted toward cell death, with increased PARK7, OPA1, ATL3, and CASP8 expression, disruption of DNA repair and cell cycle regulators (CHEK1, CDK12, MSH6, KIF2C), and decreased migration-related proteins (LAMA4, CTTN, ITGB3, A2M). Conclusions: Nano-encapsulated tarin promotes a time-dependent transition from early adaptive stress responses to apoptosis, autophagy, cell cycle disruption, and loss of migratory capacity. These findings provide novel insights into the molecular mechanisms underlying tarin antitumoral activity and support its potential as a promising therapeutic strategy against triple-negative breast cancer.

Pharmaceutics doi: 10.3390/pharmaceutics18091122

Authors: Huiya Chen Xiaoyu Zhao Minyan Zhang Yu Zhang Yuhan Huang Hanqi Zhang Min Lin Lechun Lyu Dan Xu

Background/Objectives: Bacterial wound infections remain a significant clinical burden because of prolonged healing and the increasing prevalence of multidrug-resistant bacteria. This study aimed to develop a curcumin-loaded liposomal nanosystem (Lipo-Cur) for light-triggered photodynamic antibacterial therapy and enhanced wound repair. Methods: Lipo -Cur was prepared and characterized in terms of morphology, particle size, dispersibility, drug en-capsulation, curcumin solubility, and photostability. Reactive oxygen species generation under 430 nm blue-light irradiation was evaluated. The antibacterial activity of Lipo-Cur against Staphylococcus aureus and Escherichia coli was assessed in vitro. Its effects on the proliferation, migration, and colony formation of L929 and HaCaT cells were also examined. Therapeutic efficacy and biosafety were further evaluated in a murine S. aureus-infected wound model. Results: Lipo-Cur exhibited a near-spherical morphology with an average diameter of 134.8 nm, excellent dispersibility, high drug encapsulation, and improved curcumin solubility and photostability. Under 430 nm blue-light irradiation, Lipo-Cur efficiently generated reactive oxygen species. It exhibited minimum inhibitory concentrations of 100 and 125 μg/mL against S. aureus and E. coli, respectively, and achieved bactericidal rates exceeding 95%, outperforming free curcumin. Lipo-Cur also significantly enhanced L929 and HaCaT cell proliferation, migration, and colony formation, indicating favorable biocompatibility. In the murine infect-ed-wound model, Lipo-Cur combined with blue-light irradiation (Lipo-Cur-BL+) achieved 93.76 ± 1.56% wound closure by day 12, accelerated tissue regeneration, reduced inflammation, and caused no evident systemic toxicity or hemolysis. Conclusions: These findings demonstrate that Lipo-Cur is a safe and effective photodynamic antibacterial platform with considerable potential for treating bacterially infected wounds and promoting wound healing.

Pharmaceutics doi: 10.3390/pharmaceutics18091121

Authors: Jordane S. Rodrigues Sofia O. D. Duarte Micheli de Souza Bernardes Paola Pirela Beatriz Ruivinho Filipa Ramos Rafael Parada Savino Andressa Raianny Silva Soares Pedro Henrique Gomes Araújo Fernanda Guerra Lima Medeiros Borsagli Pedro Fonte

Background: The development of sustainable biomaterials that combine imaging capability with complementary biological functions without relying on incorporated drugs or exogenous fluorophores represents an attractive strategy for multifunctional biomedical platforms. Methods: Here, carboxymethyl cellulose (CMC) hydrogels reinforced with nanographene oxide (nGO) were developed by citric acid-mediated crosslinking using nGO contents of up to 3% (w/w) and evaluated for their physicochemical, electrochemical, and biological properties. Results: nGO exhibited a nanosheet morphology, a hydrodynamic diameter of 5.0 ± 0.2 nm, and a specific surface area of approximately 650 m2 g−1. Incorporation of nGO modified the hydrogel structure and reduced water uptake within the tested formulation window. The nanocomposite hydrogels maintained high cytocompatibility, with approximately 100% HeLa cell viability after 24 h. Confocal microscopy and quantitative fluorescence-intensity analysis demonstrated enhanced fluorescence relative to untreated cells, reaching approximately eightfold higher signal intensity and supporting proof-of-concept fluorescence-assisted visualization of HeLa cells without exogenous fluorescent probes. However, because only HeLa cells were evaluated and no cancer-specific targeting ligand was incorporated, these findings do not establish diagnostic specificity. The CMC–nGO hydrogels also exhibited radical-scavenging activity of up to 96.5% and antimicrobial activity against Escherichia coli and Candida albicans, with maximum inhibition zones of 12.9 and 9.2 mm, respectively. In addition, nGO incorporation modified the electrochemical response of graphite electrodes, supporting the multifunctional character of the platform. Conclusions: Overall, CMC–nGO hydrogels provide a sustainable, drug-free, and fluorophore-free biointerface combining fluorescence-assisted cell visualization with antioxidant, antimicrobial, and electrochemical functionalities. Further photophysical characterization, comparison with non-cancerous cervical cells, rheological and mechanical evaluation, and validation in advanced biological models are required to establish their potential for future bioimaging and biosensing applications.

Pharmaceutics doi: 10.3390/pharmaceutics18091120

Authors: Chatchaya Ponsuremas Takron Chantadee Pratchaya Tipduangta Siriporn Okonogi Pimpak Phumat

Background/Objectives: Dihydromyricetin (DHM), a poorly water-soluble flavonoid with potent antioxidant and skin-rejuvenating properties, exhibits limited skin permeation. This study aimed to develop and characterize DHM-loaded in situ film-forming emulsions (FFE) using Eudragit® RS 100 as the film-forming polymer to enhance skin permeation while minimizing systemic absorption. Methods: FFE formulations, comprising a DHM-loaded lipid mixture and a Eudragit® RS 100 polymer solution, were developed using a full-factorial design of experiments varying poloxamer 407 (P407) and octyl cyanoacrylate (O60/20)/N-methyl-2-pyrrolidone (NMP) concentrations to evaluate drying time, elongation at break, water vapor transmission rate (WVTR), and dermal permeation. The optimized formulation was further characterized for physicochemical properties, chemical integrity, drug loading, and release behaviour. Results: All formulations were homogeneous and physically stable over 24 h. Drying time was positively influenced by P407 but negatively by O60/20/NMP; P407 reduced dermal permeation, while O60/20/NMP had a biphasic effect without significantly affecting elongation at break and WVTR. The optimized FFE (0.079%w/w O60/20/NMP, without P407) showed rapid drying (2.68 min), elongation >10%, and self-formed nanoscale emulsion droplets from the lipid mixture. FTIR confirmed component compatibility; XRD revealed DHM conversion from a crystalline to an amorphous state. Compared with a system without lipid, FFE exhibited significantly higher moisture content (p < 0.001), a lower swelling index (p = 0.021), comparable occlusive factor (p = 0.68), and followed Higuchi kinetic release. Conclusions: The optimized FFE demonstrated desirable film-forming properties and physicochemical compatibility, offering an innovative formulation strategy with translational potential for topical industries. However, substitution of NMP is recommended for cosmetic formulations.

Pharmaceutics doi: 10.3390/pharmaceutics18091119

Authors: Katangur Vishruth Reddy Soumalya Chakraborty Sourav Chougule Amit Pariskar Rohit Y. Sathe Ashish Dangi Prasad V. Bharatam Arvind K. Bansal

Background: In this study, two cocrystals of diclofenac acid (DCA) with the coformers theophylline (THEO) and isonicotinamide (ISNT) were prepared. Subsequently, nanococrystals were generated from these cocrystals using a top-down wet media milling approach. Methods: Critical process parameters such as milling time, milling volume, drug loading percentage, bead volume, and dispersion media were optimized to achieve the desired particle size distribution. The nanococrystals were characterized using dynamic light scattering (DLS), polarized light microscopy (PLM), differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Results: In vitro dissolution studies revealed that nanococrystals of DCA-ISNT (DE0–120 = 22.5% at pH 1.2 and DE0–120 = 58.7% at pH 4.5) and DCA-THEO (DE0–120 = 18.5% at pH 1.2 and DE0–120 = 48.2% at pH 4.5) exhibited superior dissolution performance compared to DCA nanocrystals (DE0–120 = 12.5% at pH 1.2 and DE0–120 = 39.0% at pH 4.5), with the dissolution advantage decreasing as the pH of the medium increased. The improved dissolution behaviour was a complex interplay of factors including particle size distribution, surface wetting kinetics, exposure of hydrophilic/hydrophobic functional groups during dissolution, nanococrystal microenvironmental pH, DCA’s ionization behaviour, lattice energy, and intermolecular interaction strengths. Additionally, nanococrystals exhibited a significantly higher flux rate in simultaneous gastric transfer dissolution and flux studies compared with DCA, likely due to higher apparent solubility and superior diffusion through the unstirred water layer (UWL). Pharmacokinetic studies confirmed that nanococrystals DCA-ISNT NCC (AUC0–∞ = 3062.65 ± 526.91 ng/mL·h) outperformed DCA nanocrystals (AUC0–∞ = 2352.53 ± 537.78 ng/mL·h), DCA-THEO NCC (AUC0–∞ = 2222.96 ± 151.19 ng/mL·h) and the cocrystals in terms of pharmacokinetic performance. Conclusions: The findings indicate that DCA-ISNT NCC exhibited superior pharmacokinetic performance and, together with the enhanced dissolution and flux properties of the nanococrystals, demonstrates their potential for enhanced therapeutic efficacy.

Pharmaceutics doi: 10.3390/pharmaceutics18091118

Authors: Lolita Kuršvietienė Robertas Lažauskas Inga Stanevičienė

The development of effective delivery systems remains an important challenge in modern pharmaceutical and biomedical research, driving the development of biocompatible materials with versatile functional properties. This review provides an integrated overview of chitosan-based delivery systems, focusing on chitosan nanoparticles, chitosan derivatives, and hybrid systems. Particular emphasis is placed on the relationship between the structural and functional characteristics of chitosan-based systems and their potential applications in pharmaceutical and biomedical fields. In addition to established applications, emerging approaches involving nucleic acid delivery, gene therapy, theranostics, and other biomedical applications are discussed. A further focus of this review is the consideration of critical quality attributes relevant to the development of GMP-oriented chitosan-based systems, providing a perspective that links material characteristics with quality and translational requirements. By integrating these aspects, this review highlights the versatility of chitosan as a platform for the development of innovative pharmaceutical and biomedical delivery technologies.

Pharmaceutics doi: 10.3390/pharmaceutics18091117

Authors: Su Jeong Kang Sunghyun Bong Min Jeong Jo Jae Min Lee Moon Sup Yoon Seonmin Park Yeseung Lee Yuseon Shin Hye Jin Lee Chun-Woong Park Dae Hwan Shin

Conventional in vitro drug evaluation relies largely on static concentration–response assays that fail to reproduce the dynamic pharmacokinetic (PK) profiles observed in vivo, contributing to the gap between preclinical findings and clinical outcomes. Recent advances in microphysiological systems (MPSs), particularly microfluidic organ-on-chip platforms, enable programmable concentration–time profiles that more closely mimic physiological drug exposure. These PK-informed platforms allow systematic investigation of schedule dependency, time-dependent pharmacodynamics (PD), and exposure-driven efficacy under controlled flow conditions. Spatially resolved analytical approaches further reveal heterogeneous drug penetration and metabolic responses within tissues, emphasizing the importance of spatiotemporal PK–PD coupling. Integration of multi-organ and vascularized chip systems with physiologically based pharmacokinetic (PBPK) modeling increasingly supports quantitative in vitro–in vivo translation. This review outlines how PK-informed MPSs can generate dynamic in vitro exposure and response data that inform PBPK modeling, thereby supporting quantitative in vitro–in vivo translation of drug disposition and response.

Pharmaceutics doi: 10.3390/pharmaceutics18091115

Authors: Qian Chen Hui Yang Meili Pei Yanxia Sun Yubei Li Sen Yu Xiaofeng Yang

Background: The fibrous tumor extracellular matrix (ECM), driven by cancer-associated fibroblasts (CAFs), forms a physical barrier against drugs and immune cells, yet direct CAF elimination risks promoting metastasis. Methods: In this study, we developed a locally injectable hydrogel based on synergistic dynamic covalent crosslinking (imine and boronate ester bonds), enabling instant gelation, shear thinning, and dual-pH/ROS-responsive degradation. Two types of drug-loaded nanoparticles (NPs), coated with homotypic cell membranes, were incorporated into this hydrogel. In the acidic, reactive oxygen species (ROS)-rich tumor microenvironment (TME), the system responsively releases the antifibrotic drug SIS3 to reprogram CAFs while simultaneously delivering doxorubicin (DOX) specifically to tumor cells. Biological effects were evaluated in vitro using cell cultures and in vivo in mouse models. Results: This dynamic hydrogel-based co-delivery system effectively reprograms CAFs, reduces tumor mechanical stress, breaks the fibrotic barrier, and promotes the deep infiltration of chemotherapeutics and immune cells, thereby enhancing the efficacy of chemotherapy. Conclusions: This injectable pH/ROS-responsive dynamic covalent hydrogel, loaded with CAF- and cancer cell-targeting NPs, remodels the TME, enhances drug and immune cell penetration, and offers a promising biomaterial-based strategy for cervical cancer treatment.

Pharmaceutics doi: 10.3390/pharmaceutics18091116

Authors: Melisa Kafali Emilia Mora Pinos Katharina S. Berryman Kellen Chen Geoffrey C. Gurtner Kenneth W. Liechty Carlos Zgheib

Background: Diabetic wounds are characterized by chronic inflammation, oxidative stress, impaired angiogenesis, and delayed tissue repair. Ferroptosis has emerged as a potential contributor to diabetic wound pathology; however, its relationship with impaired tissue regeneration remains incompletely understood. Objectives: We investigated cellular and transcriptional responses associated with cerium oxide nanoparticle-conjugated microRNA-146a (CNP-miR146a) treatment and whether wound repair is associated with ferroptosis-protective programs and immune–vascular communication. Methods: Diabetic excisional wounds were treated with CNP-miR146a or phosphate-buffered saline controls. Single-cell RNA sequencing was performed on wound tissues, with primary analyses focused on postoperative Day 7. Cellular composition, ferroptosis-associated programs, pseudotime trajectories, and inferred ligand–receptor communication networks were analyzed. Results: CNP-miR146a treatment was associated with transcriptional remodeling of the diabetic wound microenvironment, with myeloid cells exhibiting a prominent response. Treatment was associated with higher NRF2-related antioxidant, ferroptosis-protective, and iron-homeostasis transcriptional programs and with a repair-associated myeloid state. Pseudotime analysis identified a trajectory from monocytes toward pro-regenerative macrophages accompanied by dynamic expression of antioxidant, iron-homeostasis, and repair-associated genes. CellChat predicted increased immune–vascular communication through angiogenic and extracellular matrix-associated pathways. Endothelial cells exhibited increased NRF2-associated transcriptional programs, angiogenesis-associated gene expression, and endothelial repair markers. Conclusions: CNP-miR146a-mediated wound repair is associated with coordinated ferroptosis-protective and NRF2-related transcriptional programs, pro-regenerative myeloid states, endothelial angiogenesis-associated programs, and predicted immune–vascular communication. These findings identify ferroptosis-associated and immune–vascular transcriptional networks as candidate mechanisms of CNP-miR146a-mediated diabetic wound repair requiring further functional validation.

Pharmaceutics doi: 10.3390/pharmaceutics18091114

Authors: Yuling Wang Shujing Ren Jun Deng Dan Luo Rui Liu Yu Zhou Siyuan Chen Wei Liu

Background: Effective topical management of sensitive skin remains challenging because inadequate cutaneous delivery limits the therapeutic performance of many anti-inflammatory agents. Glycyrrhetinic acid (GA) possesses well-recognized anti-inflammatory and barrier-protective activities, yet its clinical potential is constrained by poor aqueous solubility and inefficient skin delivery. This study aimed to develop a hyaluronic acid (HA)-engineered ethosomal system to enhance the local delivery and therapeutic efficacy of GA for sensitive skin. Methods: HA-coated GA-loaded ethosomes (HAGA-ETs) were prepared by electrostatic adsorption of HA onto a cationic ethosomal template. The physicochemical properties, release behavior, storage stability, skin retention, cellular uptake, and biological activities of HAGA-ETs were systematically evaluated using TNF-α/IFN-γ-stimulated HaCaT cells and an SLS-induced 3D reconstructed skin model. Results: HAGA-ETs exhibited a mean particle size of 140.1 nm, encapsulation efficiency exceeding 95%, sustained drug release, and good storage stability. Compared with Free-GA and unmodified ethosomes, HAGA-ETs showed improved cytocompatibility, enhanced skin retention, greater keratinocyte uptake, and stronger anti-inflammatory activity. HA pre-saturation attenuated the enhanced cellular uptake of HAGA-ETs, supporting the involvement of HA receptor-mediated cellular interaction. HAGA-ETs also more effectively restored barrier-related markers, suppressed hyper-reactivity- and allergy-associated mediators, and inhibited the activation of MAPK/NF-κB, JAK1/STAT1, and TRPV1-related signaling pathways in both cellular and 3D skin models. Conclusions: HA surface engineering effectively improved the topical delivery and local therapeutic efficacy of GA by enhancing skin retention and keratinocyte interaction. HAGA-ETs represent a promising nanoplatform for the local management of sensitive skin.

Pharmaceutics doi: 10.3390/pharmaceutics18091113

Authors: Zihan Lei Hao Liang Qin Cheng Weijie Kong Yipeng Du Xueting Yao Feifei Feng Yuyan Jin Wenting Wang Haiyan Li Ming Lu Dongyang Liu Ning Shen

Background/Objectives: Meropenem pharmacokinetic variability in sepsis often leads to suboptimal exposure and therapeutic failure. Existing covariates like creatinine clearance (CLcr) only partially explain this variability. This study evaluated pyridoxic acid (PDA), an endogenous biomarker of OAT1/3 transporters, as a novel covariate to quantify active tubular secretion and explore pharmacokinetic/pharmacodynamic (PK/PD) linkages with clinical outcomes. Methods: A population PK (PopPK) model was constructed using data from a prospective septic cohort (n = 28). Subsequent exposure-response analysis was conducted in an expanded cohort (n = 49), and Monte Carlo simulations were utilized to evaluate various dosing regimens. Results: The PopPK analysis suggested that PDA may complement CLcr in characterizing meropenem clearance variability. While CLcr explained 10.7% of inter-individual variability (IIV) in clearance, the inclusion of PDA explained an additional 13.7%, reducing total IIV from 50.8% to 26.4%. Achieving a stringent target of 100%fT > 4MIC was significantly associated with a rapid decline in procalcitonin (p = 0.027), establishing a key PD endpoint. Simulations demonstrated that standard dosing (1 g q8h, 1 h infusion) is insufficient for patients with normal or augmented renal function. Target attainment was highly dependent on PDA levels. Conclusions: PDA is a valuable translational biomarker for OAT-mediated clearance. To achieve 100%fT > 4MIC, we recommend (i) 1 g q8h with 3 h infusion for patients with low CLcr and high PDA levels (MIC = 0.5 mg/L), and (ii) an intensified regimen of 2 g q8h with 3 h infusion for patients with normal CLcr and low PDA levels or high resistance risk (MIC ≥ 2 mg/L).

Pharmaceutics doi: 10.3390/pharmaceutics18091112

Authors: Yohan Seo Chul Soon Park

Major depressive disorder remains a leading cause of disability worldwide, and the limited efficacy and delayed onset of conventional antidepressants have intensified interest in the microbiota–gut–brain axis as a source of therapeutic targets. Microbiota-associated candidates—short-chain fatty acids, bile acid and tryptophan metabolites, and neuroactive amines—show mood-relevant activity, yet almost none has reached the clinic. This review reframes that gap as a delivery problem. Rather than treating entry into the central nervous system as a universal requirement, we distinguish strategies intended for local intestinal, peripheral systemic, and direct central action, and we argue that delivery is a major but not exclusive translational bottleneck. We outline the barriers these agents face—upper gastrointestinal loss, poor colonic targeting, rapid metabolite turnover, first-pass exposure, and the blood–brain barrier—and synthesize delivery strategies across two fronts. Colon-targeted systems are technically established but have been validated for non-depression indications, whereas brain-directed approaches—bacterial extracellular vesicles, detoxified membrane-coated carriers, receptor-mediated transcytosis, and intranasal routes—reach the brain mainly in selected preclinical models. We foreground a paradox: microbial extracellular vesicles are at once one of the better-documented bio-derived routes for brain exposure in preclinical studies and prominent drivers of neuroinflammation, which defines a risk–opportunity continuum. We close with a route-specific validation roadmap encompassing quantitative exposure, target engagement, chronic efficacy, and safety.

Pharmaceutics doi: 10.3390/pharmaceutics18091111

Authors: Diana Constanța Pelea Laura Maria Endres Laura Maghiar Teodor-Andrei Maghiar Mădălin Florin Ganea Csaba Nagy Corina Moisa Gabriela Ciavoi Georgiana Ioana Potra Cicalau Olimpia-Daniela Frenț Mariana Ganea

Background/Objectives: Antimicrobial resistance has renewed interest in plant extracts, essential oils and honey as topical adjuvants, yet antimicrobial potency, host-tissue safety and formulation performance are usually reported in separate bodies of literature, and the material tested is often characterized only superficially. This structured narrative review aimed to integrate these dimensions into a single decision-oriented pathway. Methods: The Web of Science Core Collection was searched using seventeen predefined strings across fourteen thematic domains, restricted to English-language articles and reviews from 2021 to 2026, with supplementary PubMed/MEDLINE searches, hand-searching, and normative documents from the issuing organizations. Results: The framework comprises seven decision domains: pre-analytical standardization of each matrix class; antibacterial and antifungal testing with matrix-specific controls; biofilm endpoints named according to what they measure; strain selection tiered from reference to clinical and resistant isolates; host safety from monolayer cytotoxicity to reconstructed human epidermis; selectivity, expressed per microorganism and extended to the resident cutaneous microbiota; and formulation performance, with permeation interpreted against a retention rather than a maximization target. Resistance under sub-inhibitory exposure forms an advanced stage. Conclusions: Progression is governed by proposed Go, Conditional Go and No-Go criteria, supported by a matrix-specific control table and a minimum reporting checklist. The framework has not been prospectively validated, and no universal selectivity threshold is proposed.

Pharmaceutics doi: 10.3390/pharmaceutics18091110

Authors: Jinhwan Shin Eu-min Kim Jun-hong Jang Seok-won Jee Sang-hoon Kim Seonggwan Yu Minguen Yoon Daniel Craig Ryan Swoyer Sandip Patel Praveen Alamuri Albert Price Rashmi Ravichandran Lauren Carter Sammaiah Pallerla

Background/Objectives: The rapid emergence of SARS-CoV-2 variants that evade neutralizing antibodies underscores the need for new antiviral biologics that integrate precise molecular design with scalable, cost-effective manufacturing. Computationally designed miniproteins targeting the spike protein’s receptor-binding domain (RBD) offer a promising alternative to monoclonal antibodies because of their small size, high thermal stability, and compatibility with microbial expression systems. Here, we detail the full development and cGMP manufacturing of IPD-52520, a novel antiviral miniprotein produced using an optimized E. coli platform. Methods: Two candidate miniproteins, a homotrimeric construct (Trimer, IPD-52520, 17 kDa) and a tandem fusion construct (Daisy, IPD-52521, 25 kDa), were evaluated in parallel through systematic optimization of strain selection, media formulation, fed-batch fermentation, inclusion-body solubilization, refolding, and chromatographic purification. The leading candidate was scaled from 5 L to 50 L under cGMP conditions, and biophysical and stability analyses were conducted to support nonclinical and Phase 1 clinical development. Results: The Trimer was selected as the lead molecule based on superior preclinical efficacy, favorable pharmacokinetics, and higher volumetric yields. The optimized process yields about 2 g/L of purified protein at over 90% purity. Scale-up from 5 L to 50 L demonstrated excellent batch consistency across six independent runs. Biophysical studies confirmed a well-folded, predominantly alpha-helical trimer (Tm = 73.4 °C; polydispersity = 1.005) with an intact primary structure and strong target binding (KD < 1 pM). Stability studies show the drug remains stable at 2–8 °C for at least 12 months, with ongoing long-term studies. Conclusions: These findings demonstrate that computationally designed antiviral miniproteins can be translated into scalable biologics and establish a platform for rapid therapeutic development against current and future pandemics.

Pharmaceutics doi: 10.3390/pharmaceutics18091109

Authors: Aurane Lecouffe Thomas Bouillet Bryan Thiroux Amélie Degraeve Anaïs-Camille Vreulx Romain Magnez Angelo Scuotto Christophe Barnier-Quer Didier Betbeder

Background/Objectives: Toxoplasma gondii is a globally distributed parasite responsible for significant morbidity in both humans and animals. VXN-Toxo, is an intranasal vaccine based on maltodextrin nanoparticles formulated with inactivated T. gondii parasites. A vaccination campaign conducted worldwide in zoological parks demonstrated high efficacy across multiple species and geographical regions. These findings suggest that VXN-Toxo may induce broad cross-reactive immunity against T. gondii strains circulating in various regions. Methods: To further characterize the immune mechanisms, we evaluated the cellular immune response induced by VXN-Toxo in C57BL/6 mice. Following vaccination, splenocytes were stimulated with antigens derived from multiple T. gondii strains, representing the major haplogroups (types I, II, III, and atypical strains). Results: ELISPOT analysis demonstrated that VXN-Toxo induced strong antigen-specific T cell responses, characterized by robust IFN-γ and IL-17 production upon stimulation with both homologous and heterologous antigens. Flow cytometry analysis further revealed the activation of both CD4+ and CD8+ T cells, with the notable presence of IFN-γ–producing CD8+ central and effector memory cells and CD4+ effector memory T cells. Conclusions: Altogether, these results indicate that VXN-Toxo induces a broad, T cell–mediated immune response with cross-reactive properties and highlight its potential as a promising vaccine candidate for both human and veterinary applications.

Pharmaceutics doi: 10.3390/pharmaceutics18091108

Authors: Lu Lu Yige Qiu Jiayu Wu Wei Dou Jing Yang Bo Zhang

RNA therapeutics offer considerable potential for cancer treatment. Their therapeutic application, however, remains limited by rapid degradation, inefficient cellular uptake, and restricted intracellular release. Extracellular vesicles (EVs) are cell-derived membrane vesicles that have been exploited as promising vehicles for drug delivery due to their high biocompatibility and low immunogenicity, whereas liposomes and lipid nanoparticles provide tunable lipid composition and efficient loading of exogenous nucleic acids. Combining these carriers has led to EV–lipid hybrid systems designed to integrate their complementary properties. This review summarizes recent advances in EV–lipid hybrids for cancer therapy and organizes the reported systems according to their structural architecture and preparation. EV–liposome fusion hybrids, EV–lipid nanoparticle hybrids, and EV membrane-integrated lipid nanocarriers are discussed in relation to their RNA-loading strategies and representative therapeutic designs. The review also examines the key processes involved in functional RNA delivery and summarizes representative applications across different cancer types. Challenges associated with safety evaluation are also discussed, together with future directions for clinical translation. Overall, EV–lipid hybrids represent a promising strategy for RNA-based cancer therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091107

Authors: Jun Liu Shuang Liu Jianxiong Wei Zifang Ding Xiaodan Yan Jin Sun Shujun Wang Shanhu Li Yuanqing Li

Background: The oral application of adenovirus is hindered by its poor in vitro storage stability and rapid degradation by gastric acid. To address this, a biomimetic oral adenovirus delivery system (CSP-AdV@LYO) was constructed based on three key properties of natural chrysanthemum sporopollenin (CSP): chemical inertness, intelligent “acid-shrinking/alkali-swelling” responsiveness, and mucosal adhesion via its spike structures, aiming to enhance oral stability and delivery efficiency. Methods: First, low-allergenic chrysanthemum pollen was screened using proteomics and a zebrafish allergy model. High-purity sporopollenin (SPO) was then extracted via an acidolysis method, followed by systematic characterization of its morphology, particle size, zeta potential, contact angle, and reversible acid-shrinking/alkali-swelling behavior. Subsequently, a CSP-AdV@LYO formulation was prepared by optimizing a cryoprotectant formulation (sucrose:gelatin = 1:1) and a vacuum loading process. Its protective and release properties were evaluated in vitro using simulated gastric and intestinal fluids, and its long-term stability was assessed. Further in vivo studies in mice assessed its intestinal colonization efficiency. The adhesion mechanism of the sporopollenin spike structures was investigated through mucosal retention experiments. Results: Mucosal retention experiments confirmed that the spike structures on the sporopollenin surface enhanced retention by approximately 3-fold through mechanical interlocking compared to smooth particles. In long-term stability tests, the viral genome copy number retention rate was improved more than 10-fold compared to the virus stock solution. The system enabled a steady and controlled release of the virus in simulated intestinal fluid, with the released virus maintaining its infectivity. In vivo studies demonstrated that CSP-AdV@LYO promoted efficient intestinal colonization and reduced acute mortality from 75% (AdV@LYO group) to 25%. Conclusions: By leveraging the unique physicochemical properties of chrysanthemum sporopollenin, this study successfully developed a biomimetic oral delivery system for live adenovirus that provides gastric acid protection, intelligent pH-responsive release, and mucosal adhesion. This system significantly enhances the oral stability and intestinal delivery efficiency of adenovirus while reducing systemic exposure risks. It offers a novel biomimetic strategy for the oral delivery of adenovirus and other biological macromolecules.

Pharmaceutics doi: 10.3390/pharmaceutics18091105

Authors: Qingqing Xu Lisong Sheng Hui Zhao Lingyun Du Jingjing Wei Huijie Zhang Tianyu Zhang Huanhuan Zhang Chunhong Zhang Rong Sun

Background/Objectives: Psoriasis is a chronic inflammatory skin disease driven by Th17/Treg imbalance. Colquhounia Root Tablet (CRT), derived from Tripterygium hypoglaucum, has shown clinical potential for psoriasis, but its mechanisms remain unclear. This study aimed to evaluate the anti-psoriatic effects of CRT and elucidate its underlying mechanisms. Methods: Anti-psoriatic activity was evaluated in an IMQ-induced psoriasis-like mouse model. Mice received oral CRT at 0.085, 0.17, or 0.35 g/kg daily from days 2 to 8. Immune-cell populations were analyzed by flow cytometry. Bone marrow-derived dendritic cells (BMDCs) were used for in vitro studies. Network pharmacology, transcriptomics, molecular docking, and experimental validation were integrated to explore the mechanisms. Results: CRT dose-dependently ameliorated psoriasiform dermatitis and reduced Th17/Treg ratio while inhibiting CD11c+MHC II+ DC activation in vivo. In vitro, CRT suppressed R848-induced BMDC maturation and inhibited p65/IκBα phosphorylation. Transcriptomic analysis revealed modulation of TNF, NF-κB, IL-17, and JAK-STAT pathways. Molecular docking predicted the strong binding of multiple CRT compounds to RELA. Conclusions: CRT exerts anti-psoriatic effects in a murine model with concurrent modulation of NF-κB-related DC maturation and Th17/Treg correction, suggesting a potential immunomodulatory mechanism requiring further causal validation.

Pharmaceutics doi: 10.3390/pharmaceutics18091106

Authors: Grzegorz Szynkaruk Julia Kerner Ahata Nelipovich Kacper Osuch Maria Miotk Agata Bartkowiak Joanna Sobiak

Background/Objectives: Saliva may provide a non-invasive alternative to blood sampling for therapeutic drug monitoring (TDM) of mycophenolic acid (MPA) in children. This study assessed MPA and, secondarily, mycophenolic acid glucuronide (MPAG) in non-centrifuged Salivette® devices to determine an acceptable interval between saliva collection and laboratory processing. Methods: Phosphate-buffered saline (PBS), artificial saliva, and saliva from five healthy adult volunteers were externally spiked with MPA and MPAG at 5 and 500 ng/mL and applied to Salivette® cotton swabs. Samples were stored for 12, 24, and 48 h at 22 °C and 6 °C and analyzed by liquid chromatography-tandem mass spectrometry. Percentage deviations from nominal concentration within ±15% were considered acceptable. Results: In PBS, both analytes met the acceptance criterion for up to 24 h under both conditions. In artificial saliva, both met the criterion for at least 24 h and, under some conditions, for 48 h. In human saliva, all volunteer-level MPA results met the criterion after 12 h at both temperatures, whereas MPAG did not consistently meet it. Mean MPA deviations across volunteers ranged from −10.8 to 1.9% at 22 °C and from −12.3 to 2.4% at 6 °C. Conclusions: MPA met the acceptance criterion after 12-h in non-centrifuged Salivette® devices, supporting the feasibility of a 12-h pre-centrifugation interval under comparable conditions. Prompt centrifugation and processing are advisable when MPAG determination is required. Confirmation using incurred post-dose saliva from pediatric patients receiving mycophenolate mofetil is required.

Pharmaceutics doi: 10.3390/pharmaceutics18091104

Authors: Tünay Karan Çağrı Çağlar Sinmez Sevgi Durna Daştan Murat Çakir Mücahit Seçme René van den Hoven

A wound is defined as disruption or destruction of tissue integrity. In order to support healing in wound management, a good wound bed free of necrotic tissue and infection is desired, but intensive chemical antiseptics will cause cell destruction and delay healing. In order for wound healing to be rapid, the nature and contamination of the wound should be taken into consideration and appropriate methods should be utilized. Today, many types of algae are frequently preferred as an alternative to medicine and are the subject of research. Since the metabolites contained in algae display several notable biological activities such as antimicrobial, anti-inflammatory, and antioxidant, they are a good option in wound treatment. Algae contain pigments, peptides, fatty acids, and polysaccharides that are crucial for wound healing. These compounds play vital roles at all stages of the healing process by accelerating cell proliferation, promoting collagen deposition, scavenging reactive oxygen species (ROS), and regulating key inflammatory cytokines. Furthermore, their unique physical and functional properties enable the development of novel bio-inspired wound dressings, hydrogels, and drug-delivery scaffolds. This review discusses the bioactive metabolites found in algae that are effective in wound healing.

Pharmaceutics doi: 10.3390/pharmaceutics18091103

Authors: Lara Marques Nuno Vale

Background/Objectives: Interindividual pharmacokinetic (PK) variability remains a daunting challenge for effective and safe drug therapy. Despite the widespread use of direct oral anticoagulants (DOACs) and calcium channel blockers (CCBs), a substantial number of adverse drug reactions have been reported for both classes. Herein, this study aimed to assess and analyze the PK variability of DOACs and CCBs across diverse clinical and demographic profiles under both single- and multiple-dose conditions. Methods: A PubMed search identified clinical PK studies reporting maximum plasma concentration (Cmax) and/or area under the concentration-time curve (AUC). The coefficient of variation (CV%) was calculated and used as a measure of PK variability. A CV% < 40% indicated low-to-moderate variability, and a CV% > 40% was defined as high variability. Results: A total of 264 studies were included following systematic screening, and the dataset was further characterized according to population features and clinical context. Among DOACs, edoxaban exhibited the lowest PK variability, whereas dabigatran showed the highest. CCBs demonstrated a broad variability spectrum, ranging from predictable agents (amlodipine and felodipine) to highly variable compounds (nisoldipine, isradipine, nimodipine, diltiazem, and verapamil). Studies evaluating drug–drug interactions, ethnicity, and specific drug-related factors were associated with increased PK variability. Conclusions: These findings suggest that fixed-dose strategies may not be universally appropriate for DOACs and CCBs, particularly in high-risk subgroups where altered exposure may lead to sub- or supratherapeutic concentrations and compromise clinical outcomes. Therefore, clinicians should avoid evaluating individual risk factors in isolation and instead consider the patient’s complete profile when selecting and adjusting pharmacotherapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091102

Authors: Hiroki Kato Xuhao Huang Erina Hilmayanti Yuichiro Kadonaga Kazuhiro Ooe Masashi Murakami Kazuya Kabayama Kazuko Kaneda-Nakashima Atsushi Toyoshima Xiaojie Yin Hiromitsu Haba Koichi Fukase

Background/Objectives: Peritoneal dissemination of malignancies leads to poor prognoses, and no effective treatment currently exists. The difficulty of treating such malignancies is likely because systemically administered drugs cannot easily target malignant cells in the abdominal cavity. High intraperitoneal drug retention, non-toxicity towards normal tissues, and successful targeting of malignant cells are important for an effective therapy. The aim of this study was to evaluate an intraperitoneally administered astatine-labeled, integrin-targeted nanodrug, mPEG(Mn:350)-S-AuNP[211At]-c[RGDfK(C)] ([211At]AuNP@PEG/RGD), with respect to its kinetics, therapeutic efficacy, and safety. Methods: C6 rat glioma cells (107), and BxPC3 (107) and PANC-1 (107) human pancreatic cancer cells were seeded intraperitoneally into nude mice, and [211At]AuNP@PEG/RGD (0.979 ± 0.194 MBq for C6 models (n = 3), 1.139 ± 0.035 MBq for BxPC3 models (n = 10), and 1.308 ± 0.039 MBq for PANC-1 models (n = 10) per mouse) or saline were intraperitoneally administered 4–7 days later. Cytotoxicity against malignant cells, pharmacokinetics after administration, therapeutic efficacy, and safety in abdominal organs were evaluated. Results: Intraperitoneally administered [211At]AuNP@PEG/RGD accumulated exclusively in the peritoneal cavity for a long period of time and showed minimal systemic diffusion through the blood. In the C6 model, the intraperitoneal tumor mass was significantly lower in the treated group compared with that of the controls (p = 0.05). For the BxPC3 (median survival time: control/treated = 41/65 days, p < 0.001) and PANC-1 (median survival time: control/treated = 19/35 days, p < 0.001) peritoneal dissemination models, survival analysis revealed that [211At]AuNP@PEG/RGD significantly prolonged overall survival. Although transient weight loss, leukopenia, and thrombocytopenia were observed at one week post-administration, a short recovery trend was evident thereafter. One month after administration, no abnormalities were found in hematological tests or histological analyses of intra-abdominal organs. Conclusions: The intraperitoneal administration of astatine-labeled integrin-targeted [211At]AuNP@PEG/RGD nanoparticles showed promising findings in terms of safety and efficacy for treating peritoneally disseminated malignant tumors.

Pharmaceutics doi: 10.3390/pharmaceutics18091101

Authors: Kaiyan Su Yan Li Dongmei Zhang Yuxuan Zhou Jianping Zhang Hongyan Zhu Yun Zhao Cheng Guo Quanjun Yang

Background: Breast cancer remains the most common malignant tumor among women worldwide. Although conventional treatments including surgery, chemotherapy, and radiotherapy are effective, they are confronted with challenges such as tumor heterogeneity, systemic toxicity, and recurrence driven by drug resistance. To overcome these limitations, natural products have emerged as promising therapeutic alternatives owing to their multi-target efficacy and favorable biocompatibility. However, their clinical translation is still hindered by poor chemical stability, low aqueous solubility, and inadequate bioavailability. Nanodelivery systems offer a transformative solution by enhancing bioavailability and enabling precision targeting through the enhanced permeation and retention effect, thereby widening the therapeutic window and minimizing off-target toxicity. Purpose: This review evaluates diverse nanoparticle-based delivery systems and their targeting mechanisms in natural product-based breast cancer therapy. By examining inherent advantages and translational challenges, this analysis provides critical insights into the clinical development and application of these nanoformulations. Methods: A systematic literature search was performed in PubMed, ScienceDirect, Springer, Taylor & Francis, and Web of Science to identify relevant studies on natural product-based nanoformulations for breast cancer therapy. Results: Natural products exert anti-breast cancer effects through mechanisms such as inducing apoptosis, arresting the cell cycle, inhibiting invasion and metastasis, suppressing angiogenesis, and regulating autophagy. To overcome clinical hurdles, three complementary targeting strategies have been developed: passive, active, and stimuli-responsive targeting. These advances are shifting nanomedicines toward active precision therapy, markedly improving the therapeutic index. With multiple formulations already approved or in clinical pipelines, this field is rapidly progressing from laboratory research to clinical implementation. Conclusions: Natural products possess potent anti-breast cancer effects, and the application of nanodelivery technology effectively overcomes their inherent limitations of poor stability and low bioavailability. Although preliminary findings are promising, large-scale, randomized controlled clinical trials are urgently needed to systematically evaluate their safety, efficacy, and practical potential for clinical translation in breast cancer management.

Pharmaceutics doi: 10.3390/pharmaceutics18091100

Authors: Abhishek Dadhich Vikas Sharma Shivika Sharma Sweta Bawari Iyyakkannu Sivanesan

Type 2 diabetes mellitus (T2DM) remains a global health crisis, with nearly 589 million adults currently affected and projections pointing toward 853 million by 2050. Despite an expanding pharmacological armamentarium, a significant proportion of patients fail to achieve adequate glycaemic control, and the limitations of existing therapies, including adverse effects, cost, and limited accessibility, underscore the compelling need for novel therapeutic approaches. Phytochemicals such as curcumin, berberine, quercetin, resveratrol, and epigallocatechin gallate possess well-documented antidiabetic activity, operating through the PI3K/Akt, AMPK (activated protein kinase), NF-κB/JNK (nuclear factor kappa-B), and GLP-1R (glucagon-like peptide-1) signalling axes to improve insulin sensitivity, suppress gluconeogenesis, protect pancreatic beta-cells, and attenuate chronic metabolic inflammation. However, their clinical utility has been fundamentally constrained by poor oral bioavailability arising from low aqueous solubility, gastrointestinal instability, extensive first-pass metabolism, and P-glycoprotein-mediated efflux. Advanced drug delivery systems, including liposomes, solid lipid nanoparticles (SLN), nanostructured lipid carriers, PLGA (Poly (lactic-co-glycolic acid)) and chitosan nanoparticles, nanoemulsions, self-nanoemulsifying drug delivery systems, and phytosomes have demonstrated the capacity to overcome these barriers, achieving five- to ten-fold improvements in systemic bioavailability and substantially enhanced antidiabetic efficacy in preclinical models. Emerging mechanistic evidence further positions gut microbiota modulation and epigenetic reprogramming as additional therapeutic axes through which nano-encapsulated phytochemicals may exert durable metabolic benefits. Nonetheless, critical translational challenges persist, encompassing nanotoxicological risks, herb–drug interactions, the absence of harmonised regulatory frameworks for nano-phytomedicine products, phytochemical raw material variability, and formidable technical and economic barriers to scalable nanoparticle manufacturing. This review synthesises the current mechanistic, formulation, and clinical evidence within a unified analytical framework and identifies the strategic priorities of rigorous clinical development, regulatory clarity, and manufacturing standardisation required to translate nano-phytomedicine science into evidence-based T2DM therapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18091099

Authors: Emily Cherny Tamara Minko

Background/Objectives: Intranasal delivery is a promising noninvasive approach to enhance central nervous system drug delivery in Parkinson’s disease (PD), potentially bypassing the blood–brain barrier and minimizing gastrointestinal side effects. This review summarizes clinical trials and translational evidence for intranasal PD therapies by treatment options and intent, highlighting key challenges in efficacy, pharmacokinetics, and safety. Methods: We reviewed human clinical trials, preclinical and pilot studies, and pharmacokinetic investigations of intranasal therapies for PD. Comparisons to established treatments were included in context. Only PD-specific clinical studies were analyzed. Therapies were grouped by rescue, antioxidant/metabolic/hormonal, and biologic or cell-based approaches, with continuous-delivery systems reviewed separately. Results: Intranasal rescue therapies, such as apomorphine, provide rapid improvement during OFF episodes (periods when the effects of PD medications fade and symptoms reappear), but tolerability issues and nasal irritation limit usage. Early-phase studies suggest intranasal delivery enables quick symptom relief and favorable pharmacokinetics, though most trials are small and focus on feasibility. New approaches include antioxidants, neurotrophic factors, gene therapy, and cell-based methods, with advanced formulations enhancing nasal retention and brain uptake. However, more translational evidence and long-term safety data are needed. Conclusions: Intranasal therapies for PD offer rapid rescue and expand options beyond standard drugs. Large, well-designed trials are needed to confirm efficacy, long-term safety, and optimal formulations. Intranasal delivery remains an emerging but potentially transformative strategy requiring further clinical research.

Pharmaceutics doi: 10.3390/pharmaceutics18091098

Authors: Shery Jacob Namitha Raichel Varkey Sai H. S. Boddu Jigar N. Shah Ibrahim Mustafa Abdi Rekha Rao

Extracellular vesicles (EVs) are naturally occurring nanoscale carriers that have gained attention as next-generation platforms for diagnostics, site-specific drug delivery, and tissue engineering owing to their high biocompatibility, minimal immunogenicity, and capacity to transport diverse bioactive cargo across biological barriers. This review discusses the classification, biogenesis, molecular constituents, and therapeutic properties of the major EV subtypes such as exosomes, microvesicles, and apoptotic bodies. It also highlights recent advances in EV engineering for cancer treatment, emphasizing immune modulation and targeted therapeutic delivery. Particular attention is given to plant-derived EVs, which have shown promise as scalable, low-toxicity nanotherapeutics with inherent bioactivity and effective drug delivery potential. Selected preclinical studies, recent patents, and ongoing clinical trials are also summarized, providing an up-to-date perspective on the clinical translation of EV-based technologies. Current challenges in EV isolation, characterization, scalable manufacturing, cargo loading, standardization, and regulatory approval, along with future directions for clinical translation, are summarized. Collectively, this review summarizes the growing applicability of EVs as next-generation platforms for precision medicine, targeted drug delivery, and regenerative therapies while identifying the major obstacles that must be addressed to facilitate their successful clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18091097

Authors: Adnan Amin Touseef Nawaz Oberdan Oliveira Ferreira Mozaniel Santana de Oliveira

Targeted protein degradation (TPD) represents a whole new paradigm in cell-level therapeutic design, with its ability to remove target proteins, normally through the endogenous proteasomal, lysosomal, or autophagic systems, rather than the traditional occupancy-driven inhibition approach. But the clinical efficacy of degraders is becoming more restricted based on delivery rather than efficacy only. Many proteolysis-targeting chimeras and new proximity-inducing systems have low solubility, are impermeable, are pharmacodynamically complicated, lack tissue selectivity, and cannot fully access the intracellular space. Nanomedicine and PD platforms could provide strategies not only to overcome these challenges, but also to provide other advantages, including enhancing exposure to degraders, biodistribution, controlled release, and context-dependent activation. This critical review is an outline of all lipid, polymeric, inorganic, biomimetic, targeted, activatable, and self-assembling delivery systems for TPD. We assess compositional considerations, in vitro and in vivo evidence, challenges for translation, and clinical endpoints required to support delivery-enabled degradation. Trusted TPD therapeutics need to relate different aspects of their design, such as degrader chemistry, carrier structure, disease biology, and pharmacodynamic biomarkers, to one another. Further investigations are needed to establish intact delivery of the degrader to the target, target depletion in relevant tissues, prolonged pharmacodynamics, favorable safety, and compelling therapeutic benefit relative to free degraders or traditional inhibitors. Thus, it is important to view delivery not simply as an additional step during formulation but as a design principle necessary for the reliable clinical outcome of degradation medicine.

Pharmaceutics doi: 10.3390/pharmaceutics18091096

Authors: Ludwika Piwowarczyk Jagoda Szkudlarek Dariusz T. Mlynarczyk Szymon Tomczak Violetta Krajka-Kuźniak Aleksandra Majchrzak-Celińska Robert Kleszcz Ewelina Musielak Mateusz de Mezer Emilia Cicha Gabriela Anglart Anna Jelińska

Background/Objectives: Psoriasis is an incurable chronic immune-mediated inflammatory skin disorder for which effective topical treatment remains essential. Given the reported anti-inflammatory and dermatological potential of 20-hydroxyecdysone (20HE), this study evaluated its biological activity in normal and psoriatic keratinocytes and developed stable 20HE-loaded niosomes intended for potential future dermal delivery. Methods: The biological activity of 20HE was assessed in human epidermal keratinocytes (HEKs) and psoriasis-patient-derived keratinocytes (PHEKs) after 48 h of treatment by quantifying selected cytokines. Niosomes were prepared by thin-film hydration, rehydrated with water or citrate buffer, and sonicated. The formulations were characterized for particle size, polydispersity index, zeta potential, pH, encapsulation efficiency (EE), and 21-day storage stability. Potential in vivo toxicity was assessed in Danio rerio embryos by monitoring mortality, sublethal developmental changes, and locomotor behavior after exposure to nanoformulations for up to 96 h post-fertilization. Results: 20HE showed selective anti-inflammatory activity in PHEK cells, reducing IL-4, IL-17A, and IL-22 levels by 20–34% under elevated inflammatory conditions, without affecting healthy HEK cells. The formulations had particle sizes below 300 nm and acceptable polydispersity and zeta potential values, and they remained stable during storage. In the Danio rerio embryo model, the 1A/water caused sublethal developmental abnormalities at 96 hpf, whereas 1C/water (with 20HE) was associated only with mild pericardial edema after prolonged exposure. No significant behavioral alterations were observed across the tested concentration range, and the Lowest Observed Effect Concentration for 20HE was 20 μM. Conclusions: Free 20HE reduced the levels of IL-4, IL-17A, and IL-22 in PHEK cells, indicating its modulatory effect on cytokine production under the investigated inflammatory conditions. The developed 20HE-loaded niosomes showed favorable physicochemical properties and short-term stability, supporting their potential as candidate nanocarrier systems for future dermal application studies. The findings of the present study provide a basis for further investigation, particularly studies on drug release from the niosomal formulation and skin permeation from the final pharmaceutical dosage form.

Pharmaceutics doi: 10.3390/pharmaceutics18091095

Authors: Sara Hasan Seyedeh Negin Kassaee Derek J. Richard Nazrul Islam Emad L. Izake

Background/Objectives: The recent advances in pulmonary delivery have shifted the paradigm to the development of inhalable drug-loaded polysaccharide particles that can be positioned at the respiratory interface while reducing the systemic exposure. Quercetin has broad anticancer activity but remains difficult to translate because of its poor aqueous solubility, limited bioavailability and rapid metabolic loss. Here, we report quercetin-loaded chitosan/Salvia hispanica polysaccharide nanoparticles as a natural polyelectrolyte nanocarrier for pulmonary delivery in lung cancer. Methods: Central composite design (CCD) and artificial neural network (ANN) models were used for mapping the factors to responses. Results: The models displayed a higher predictive accuracy and optimization reliability for identifying the optimized formulation. The optimized nanoparticles showed a quasi-spherical morphology (mean hydrodynamic diameter = 331 ± 14.34 nm) with cationic ζ-potential of +36.3 ± 2.56 mV and polydispersity index of 0.15. The optimized formulation showed acceptable powder-flow characteristics, an encapsulation efficiency of 73.8 ± 0.73%, drug loading of 14.20 ± 0.22%, and biphasic release with sustained quercetin release over 48 h. In A549 and H460 cell lines, nanoencapsulation increased the antiproliferative effect of quercetin relative to the free compound, yielding lower IC50 values after 48 h of exposure. The nanoparticles showed greater suppression of wound closure, increased reactive oxygen species fluorescence and clear cellular uptake. Blank nanoparticles produced only limited effects. Conclusions: These results indicate that CCD/ANN-guided chitosan/Salvia hispanica polysaccharide nanoparticles provide a promising nanodelivery tool for quercetin delivery and enhanced in vitro activity in lung cancer cells, while the observed aerosol performance advocates further investigation of their pulmonary delivery potential.

Pharmaceutics doi: 10.3390/pharmaceutics18091094

Authors: Yongfeng Yang Tingting Song Kaili Huang Hong Huang Maoyuan Zhao Yi Li

Gene therapy and genome editing increasingly depend on the safe, effective, and cell-selective delivery of nucleic acids and protein–nucleic acid complexes. Although viral vectors remain important for applications requiring durable gene expression, non-viral vectors offer advantages in cargo capacity, modularity, transient expression, potential repeat dosing, and avoidance of vector–genome integration. Lipid nanoparticles (LNPs), polymeric nanoparticles, inorganic nanomaterials, extracellular vesicles (EVs), and biomimetic hybrid systems have consequently become central platforms for delivery of siRNA, mRNA, plasmid DNA, antisense oligonucleotides, and CRISPR-based genome editors. Among these, ionizable LNPs are currently the most clinically mature non-viral technology, supported by the clinical success of siRNA therapeutics and mRNA vaccines, as well as the emergence of in vivo CRISPR therapies. Nevertheless, efficient endosomal escape, cell-type-selective targeting, extrahepatic delivery, and repeat-dose tolerability remain substantial barriers. Polymeric vectors provide broad chemical tunability, allowing adjustment of charge density, degradability, stimulus responsiveness, intracellular trafficking, and cargo release. However, toxicity and batch-to-batch reproducibility remain key concerns. EVs provide a biologically derived alternative with favorable membrane interfaces and potential advantages for protein and ribonucleoprotein delivery, but their clinical translation is constrained by heterogeneity, loading efficiency, product characterization, and scalable manufacturing. This review summarizes recent advances in non-viral gene-delivery platforms, compares their strengths and limitations, and discusses future directions in cell-selective delivery, endosomal escape, transient delivery of genome-editing machinery, engineered EVs, hybrid vectors, and manufacturing-oriented development. The field is transitioning from organ-level delivery toward delivery of the correct payload to the correct cell type at a clinically relevant exposure and safety margin.

Pharmaceutics doi: 10.3390/pharmaceutics18091093

Authors: Jahanvi Patel Michael Stolinski Anil Vangala

Insulin delivery via the buccal route has shown long-standing promise for diabetes therapy, but clinical translation remains limited. Most systems address only part of the delivery problem, whereas a viable product must simultaneously provide adequate insulin residence time, effective mucus and epithelial permeation, stability, dose consistency, safety, and manufacturability. This review examines the biological and formulation barriers limiting buccal insulin delivery and compares major platform types, including mucoadhesive films and patches, nanocarrier-based systems, deformable vesicles, chemistry-led permeation approaches, and device-enabled strategies. Stability, alongside limited permeability, is considered a key barrier, while aggregation and excipient-related instability remain insufficiently addressed. This review also discusses why promising preclinical findings often fail to translate, exhibiting low bioavailability, variability, safety concerns, and scale-up challenges recurring across platforms. Overall, further progress in buccal insulin delivery will depend on formulation strategies that better integrate permeation enhancement, stability preservation, and translational feasibility.

Pharmaceutics doi: 10.3390/pharmaceutics18091092

Authors: Anna Simonsson Nicklas Bunta Sundin Tobias Bramer Alex Wimbush Göran Alderborn

Background/Objectives: Packing, flow, and aerosolization properties of a series of binary adhesive mixtures containing micronized or spray-dried drugs were investigated, and the relationships between these blend properties and the blend structure were studied. Methods: Micronized or spray-dried terbutaline sulfate and salbutamol sulfate were used as model drugs, and an α-lactose powder was used as the carrier. Binary mixtures with drug loads ranging from 2 to 20% were prepared. The bulk density, compressibility, permeability, and shearing properties of the carrier powder and mixtures were determined, along with the in vitro aerosolization propensity of the mixtures using two types of inhalers. Imaging of the mixtures was used to assess the blend structure. Conclusions: The particle engineering method gave differences in particle crystallinity and morphology. The development of the adhesive layer with drug load was broadly consistent with the blend state concept. Spray-dried particles, however, exhibited a higher propensity to localize within surface cavities on the carrier and produced a more voluminous enveloped adhesive layer. The spray-dried particles gave a higher bulk density, a lower Hausner ratio, comparable shear strength, and a lower angle of internal friction. Aerosolization performance, including metrics such as fine particle fraction (FPF), depended on inhaler design; nevertheless, for both inhalers, aerosolization behavior was influenced by blend state and physicochemical properties of the drug. At low drug loads, spray-dried particles dispersed to a lower degree, while at high drug loads, the dispersion performance of the two particle types converged. For example, at an intermediate drug load of 7.4%, the FPF was about 20% for the spray-dried drugs and about 30% for the micronized drugs using the Screenhaler device, while the corresponding FPF:s were about 25% and 50% for spray-dried drugs and 40% and 55% for crystalline drugs using the Monodose inhaler. Overall, the physical characteristics of the drug particles were found to influence the structural evolution of the blends, as well as their mechanical and aerosolization properties.

Pharmaceutics doi: 10.3390/pharmaceutics18091091

Authors: Lania Ali Hussein Alexandra Nedic Andrejs Sitovs Valentyn Mohylyuk

Although oral peptide drug administration is the most desirable for both patients and healthcare systems, it is a major pharmaceutical challenge. Typically, less than 1% of orally administered peptide therapeutics reaches the systemic circulation. Modern research has focused on overcoming the physiological barriers responsible for the poor peptide drug bioavailability, while little attention has been given to understanding how measurable absorption occurs despite the barriers. This review aimed to explore the physiological factors that influence oral peptide absorption, such as enzymatic degradation and pH effects, motility, microbiota, transporters, tight junctions, and the effect of inflammation. The available data indicate that these factors function not only as barriers but may also create limited opportunities for peptide uptake. Enzymatic degradation and microbial metabolism reduce the availability of intact peptide drugs yet may generate biologically relevant fragments. Tight junctions are generally regarded as restrictive barriers but are shown to exhibit dynamic regulations that may permit transient paracellular transport. Similarly, gastrointestinal motility, microbiota, and inflammatory processes influence epithelial permeability and duration of peptide exposure both to absorptive surfaces and to degradative enzymes. In contrast, transporter-mediated uptake appears unlikely to account for the absorption of intact peptide drugs due to their substrate size limitations. Together, the findings suggest that measurable oral peptide absorption is unlikely to result from a single dominant pathway but rather from the cumulative contribution of the mechanisms. Improved understanding of these endogenous mechanisms may open opportunities to enhance oral peptide bioavailability and support the development of more effective therapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18091090

Authors: Chao Fang Lixin Xie Daihan Xie Jingting Yin Shiji Zhang Yiling Gan Xiaodong Hou Fanlei Kong Yu Huo Xiuli Liu

Introduction: Acute lung injury (ALI) is characterized by severe inflammation and oxidative stress. However, effective pharmacological interventions for ALI remain limited. Carnosine, an endogenous dipeptide known for its redox-regulating and immunomodulatory activities, has demonstrated promising protective effects. In the present study, inhalable carnosine-loaded liposomes (Cn-Ls) were developed to enhance pulmonary delivery and achieve localized treatment in a lipopolysaccharide (LPS)-induced ALI model in mice. Methods: Cn-Ls were prepared and systematically evaluated for their morphology, stability, drug-loading capacity, and release kinetics. In vitro assays were performed to evaluate their cytocompatibility, antioxidant activity, and effects on LPS-induced reactive oxygen species (ROS) generation. In an ALI model, inhaled Cn-Ls were administered to assess pulmonary retention and therapeutic efficacy, including lung inflammation, oxidative stress, circulating levels of C-reactive protein (CRP), tumor necrosis factor (TNF)-α, interleukin (IL)-6, lung architecture, and respiratory function. Results: Encapsulation of carnosine within liposomes markedly prolonged its pulmonary retention (t1/2 = 1.7 h vs. 1.0 h for free carnosine), providing a more sustained lung-retentive delivery profile for up to 12 h. In vitro assays showed that Cn-Ls had excellent cytocompatibility, reduced cell death, exhibited potent antioxidant activity, and effectively suppressed LPS-induced ROS generation. In an ALI model, inhaled Cn-Ls markedly mitigated lung inflammation and oxidative stress, reduced circulating levels of CRP, TNF-α, and IL-6, preserved lung architecture, and improved respiratory function. Compared with free carnosine, Cn-Ls exhibited enhanced pulmonary retention and superior therapeutic efficacy. Conclusions: These results identified inhalable Cn-Ls as a potential nanotherapeutic approach for targeted ALI management and provided a foundation for further translational development. However, additional investigations are required to assess the long-term safety of Cn-Ls, optimize formulation stability and scalability, and further elucidate the underlying therapeutic mechanisms before clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18091089

Authors: Smita Salunke Chi Kin Anthony Chan Sifan Hu

Background/Objectives: The administration of medicines via enteral feeding tubes presents unique challenges, including dose loss, tube blockage, and user-dependent variability. Although Quality by Design (QbD) frameworks provide a structured approach to product quality, administration-related quality attributes are rarely assessed in a systematic manner during pharmaceutical development. A conceptual framework, termed the Quality Target Administration Profile (QTAP), was developed to extend QbD principles to the administration stage. Methods: The framework comprises three phases: administration context characterization, administration attribute identification, and risk-based determination of Critical Administration Attributes (CAAs). QTAP was applied to three representative EFT administration scenarios: a ready-to-use oral suspension, a crushed tablet, and an oral solution. Results: Application of the framework identified 18–32 administration attributes across the three representative scenarios and prioritized 5–6 CAAs per scenario according to clinical impact, variability risk, and human-factor considerations. Administration quality requirements varied across scenarios, reflecting differences in administration context, including dosage form, patient population, clinical setting, and user characteristics. QTAP provides a structured approach for identifying and prioritizing administration-related quality considerations within pharmaceutical development. Conclusions: As a conceptual framework, QTAP is intended to complement rather than replace existing QbD approaches. Future prospective validation, including human-factor studies and real-world evaluations, is needed to establish its practical utility and predictive value.

Pharmaceutics doi: 10.3390/pharmaceutics18091088

Authors: Hao-Yuan Lee Yu-Chia Chang Chyi-Liang Chen Shu-Hua Ko Yu-Ling Huang Shang-Po Shen Yu-Lung Hsu Wen-Yuan Lee Hung-Chih Lin

Background/Objectives: Autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD) substantially affect quality of life. Previous studies have not evaluated whether the associations between early-life antibiotic exposure and the risks of ASD and ADHD differ according to the antibiotic agents administered during infancy. Methods: This nationwide case–control study used Taiwan’s National Health Insurance Research Database, Birth Reporting Database, and Maternal and Child Health Database. The cohort included 1,693,718 term neonates born between 2004 and 2015 and followed through to 2022. For ASD (n = 7265) and ADHD (n = 39,056) cases, antibiotic-exposed children were matched 1:1 with unexposed children based on sex, gestational age, birth weight, and birth year. Adjusted hazard ratios (aHRs) and 95% confidence intervals (CIs) were estimated using multivariable Cox proportional hazards models. Results: Oral antibiotic exposure was associated with lower ASD but higher ADHD risk, while parenteral exposure was associated with increased risks of both outcomes. Amoxicillin and erythromycin exposure during the first year of life were associated with lower ASD risk, whereas ampicillin exposure at 4–12 months was associated with higher ASD risk (all p ≤ 0.033). For ADHD, exposure to amoxicillin/clavulanic acid, cefixime, ampicillin, and ampicillin/sulbactam was associated with increased risk, whereas erythromycin, cephalexin, and sulfamethoxazole-trimethoprim were associated with reduced risk (all p < 0.047). Conclusions: Associations between early-life antibiotic exposure and the risks of ASD and ADHD varied according to antibiotic agents, route of administration, and timing of exposure. These findings may help explain inconsistencies among previous studies and inform antibiotic prescribing during infancy.

Pharmaceutics doi: 10.3390/pharmaceutics18091084

Authors: Luigi Ruccolo Aleksandra Evangelista Francesco Andresini Rossella Dorati Ida Genta Marco Benazzo Pietro Canzi Elena Carlotto Bice Conti Silvia Pisani

Background/Objectives: Shape-memory biodegradable scaffolds (4D scaffolds) represent promising platforms for minimally invasive tissue engineering and localized drug delivery. This study investigated how two different fabrication techniques, electrospinning (ES) and extrusion-based direct ink writing (DIW), influence the structural, thermal, mechanical, shape-memory, and drug-release properties of poly(L-lactide-co-caprolactone) (PLA/PCL 70:30) scaffolds loaded with dexamethasone (DXM). Methods: DXM-loaded PLA/PCL 70:30 scaffolds were fabricated by ES and DIW. The resulting matrices were characterized in terms of morphology, mass, thickness, drug-loading efficiency, thermal properties by differential scanning calorimetry, shape-memory performance, tensile mechanical properties, and in vitro DXM release. Results: Both fabrication techniques produced DXM-loaded matrices with comparable mass and thickness and high loading efficiencies (>82%). Glass transition temperatures ranged between 33 and 39 °C, supporting thermally induced shape recovery under physiologically relevant conditions, while ES processing was associated with higher polymer crystallinity. All scaffolds exhibited shape-memory behavior, with recovery ratios exceeding 90%. ES scaffolds displayed a microporous nanofibrous architecture, whereas DIW scaffolds showed a more open and highly porous structure. These morphological differences were reflected in their mechanical behavior: ES scaffolds exhibited higher tensile strength (up to 16.5 MPa vs. 1.9 MPa) and elongation at break (up to 320% vs. 243%). Drug-release profiles were also fabrication-dependent, with ES scaffolds reaching a plateau at approximately 80% DXM release, whereas DIW scaffolds showed near-complete release within 48 h. Conclusions: Both fabrication approaches preserved the thermoresponsive shape-memory behavior of PLA/PCL 70:30 but generated distinct scaffold architectures that strongly influenced mechanical performance and DXM-release kinetics.

Pharmaceutics doi: 10.3390/pharmaceutics18091087

Authors: Pierpaolo Palermo Davide De Angelis Elisa Sgarbi Irene Bassanetti Michael M. Tunney Dimitrios A. Lamprou

Backgorund/Objectives: Respiratory diseases represent a substantial global health burden and require effective localised pulmonary delivery strategies, particularly for poorly water-soluble therapeutic molecules. Nanoparticle-based drug delivery systems, especially those manufactured using microfluidics, have emerged as promising approaches to overcome pulmonary barriers, enhance local drug retention, and reduce systemic side effects. Among these nanocarriers, solid lipid nanoparticles (SLNs) and solid hybrid nanoparticles (SHNs) combine biocompatibility with controlled release and improved formulation stability. Methods: In this study, SLNs and lipid–chitosan SHNs were developed using microfluidic technology as candidate platforms for pulmonary drug delivery, with Cyclosporine A (CyA) used as a model hydrophobic cyclic peptide. Nanocarriers were produced using 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol as lipids, with low-molecular-weight chitosan incorporated to obtain hybrid systems. Physicochemical properties were evaluated using dynamic light scattering (DLS) and ζ potential measurements, while morphology and structural organisation were investigated using transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). Results: The microfluidic approach enabled the production of nanoparticles with controlled sizes below 200 nm, narrow size distributions, and good reproducibility. In addition, the SHNs exhibited a positive surface charge, high encapsulation efficiency (~80%), and good colloidal and thermal stability. In vitro release studies showed an initial burst release followed by sustained CyA release, reaching approximately 94% cumulative release within 6 h. The Korsmeyer–Peppas model was used as the standard kinetic model. No blank nanoparticles were used as controls in the EE and release assay. Conclusions: Overall, these findings support further investigation of microfluidic-produced lipid and hybrid nanoparticles as candidate platforms for pulmonary drug delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18091086

Authors: Jessica R. Boulos Karen E. Russi Jordan Kinnitt Daphne Gomez Escudero Tyler Willis Jorrian Abadeer Emalee Mann Aaron Cristina Anderson Angela Alexander-Bryant

Background/Objectives: RNA interference (RNAi) represents a promising therapeutic approach for silencing oncogenes involved in cancer progression by utilizing small interfering RNA (siRNA). However, siRNA requires an efficient delivery system to overcome cellular uptake and endosomal escape barriers. This study aimed to evaluate a multifunctional tandem peptide, GE11-599, designed to enhance the targeted delivery of siRNA and maintain its bioactivity in glioblastoma (GBM) cells. Methods: The GE11-599 peptide, consisting of an EGFR-targeting GE11 motif and a 599 fusogenic domain, was complexed with siRNA via electrostatic interactions to form nanoparticles. We assessed nanoparticle physicochemical properties, protection of siRNA from serum and RNase degradation, and cellular uptake in two GBM cell lines (U118MG and U87MG). Mechanistic studies evaluated receptor-mediated endocytosis and the subsequent escape from endosomes. Functional assays quantified STAT3 gene silencing and downstream effects on cell migration following treatment with GE11-599–siSTAT3 complexes. Results: GE11-599 formed positively charged, monodisperse nanoparticles capable of protecting siRNA from degradation. The tandem peptide significantly enhanced cellular internalization through EGFR-mediated endocytosis and facilitated endosomal escape of siRNA. Treatment with GE11-599–siSTAT3 resulted in robust gene silencing, achieving up to an 80% reduction in STAT3 mRNA expression. Downstream functional assessment showed a 40% decrease in migration in GBM cells treated with GE11-599–siSTAT3 complexes. Conclusions: The GE11-599 tandem peptide effectively enhances cell-specific internalization and endosomal escape of siRNA in GBM cells, resulting in increased siRNA bioactivity and functional gene silencing. These findings support GE11-599 as a promising siRNA delivery platform for targeting EGFR-expressing cancers.

Pharmaceutics doi: 10.3390/pharmaceutics18091085

Authors: Yitong Yang Dandan Guo Wei Chen Binbin Sun Mengyu Qiu Kai Wang Wenbo Dou Kang Wang Zhanjiang Zhang Shuying Feng

Background: Methotrexate (MTX) is a well-established therapeutic agent for psoriasis owing to its anti-inflammatory and immunomodulatory effects. However, its clinical use is limited by insufficient local accumulation in skin lesions and the potential risk of systemic exposure. Dunaliella salina-derived exosome (DsEXO) has favorable biocompatibility, low immunogenicity and potential skin delivery capacity, making it a promising natural nanocarrier for topical MTX delivery. This study aimed to construct Dunaliella salina-derived exosome loaded with methotrexate (DsEXO@MTX) and evaluate its therapeutic efficacy and potential mechanisms in psoriasis-like skin inflammation. Methods: DsEXO@MTX was prepared and characterized in terms of morphology, particle size, surface charge and drug-loading capacity. Cellular uptake, skin retention and tissue distribution were evaluated using fluorescence imaging and skin section analysis. Therapeutic efficacy was evaluated in an imiquimod-induced psoriasis-like mouse model by clinical scoring, histopathological examination, spleen index measurement and Ki-67 immunofluorescence staining. STAT3 phosphorylation and Th17/Treg differentiation were further examined to explore the potential immunomodulatory mechanism. Results: DsEXO@MTX exhibited a relatively uniform particle size distribution and drug-loading capacity. In vivo fluorescence imaging and skin section analysis showed that DsEXO@MTX enhanced local skin retention and promoted fluorescence distribution in epidermal and dermal regions. It significantly alleviated IMQ-induced erythema, scaling, epidermal thickening, inflammatory infiltration, splenomegaly and abnormal keratinocyte proliferation in psoriasis-like mice. Mechanistically, DsEXO@MTX reduced STAT3 phosphorylation and modulated Th17/Treg differentiation, suggesting restoration of immune balance in psoriatic inflammation. Conclusions: DsEXO@MTX represents a natural exosome-like nanovesicle-mediated topical MTX delivery system that improves local drug delivery and enhances therapeutic efficacy in IMQ-induced psoriasis-like skin inflammation, particularly when administered topically. These findings provide a potential strategy for safer and more efficient local treatment of psoriasis.

Pharmaceutics doi: 10.3390/pharmaceutics18091083

Authors: Adnan Murad Bhayo Ying Li Alaa R. Aboushanab Junyi Lin Ashkan Hassankhanirad Wei Li Jingjing Sun

Triple-negative breast cancer (TNBC) is an aggressive and heterogeneous subtype lacking effective targeted therapies. Its tumor microenvironment (TME) exhibits distinct features, including acidity, redox imbalance, elevated reactive oxygen species (ROS), and hypoxia, which provide exploitable triggers for targeted drug delivery. Stimuli-responsive polymeric nanocarriers have emerged as promising platforms that enable spatiotemporally controlled and site-specific therapeutic release in response to these endogenous cues, as well as exogenous stimuli such as temperature and light. These systems improve drug accumulation, penetration, and therapeutic efficacy while reducing systemic toxicity. Unlike previous reviews that broadly discuss nanocarriers in cancer therapy, this review focuses on the structure–function relationships of TME-responsive polymeric systems in TNBC and their translational limitations. We summarize recent advances in pH-, redox-, ROS-, hypoxia-, photo- and temperature-responsive polymers, highlighting their design strategies and therapeutic applications. Key challenges, including stimulus heterogeneity, limited in vivo validation, and clinical translation barriers, are also discussed. This review provides a concise framework for the rational design of programmable, multi-responsive polymeric nanomedicines for TNBC therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18091082

Authors: Abdullah A. Assiri

Oral targeted therapies now constitute a substantial and growing proportion of anticancer drug therapy, shifting administration from the controlled intravenous setting to patient-managed oral therapy in the outpatient setting, where systemic exposure depends on factors that parenteral therapy largely bypasses. Food effects, gastric pH, first-pass metabolism, transporter activity, organ function, concomitant medications and adherence contribute to variability in exposure, and many oral anticancer agents have narrow therapeutic indices in which modest exposure changes carry clinical consequence. This review synthesizes the pharmacokinetic determinants of oral anticancer drug exposure across twenty-seven exemplar agents spanning the main mechanistic classes and translates them into actionable pharmacy practice. Its contribution is a cross-class agent-level comparison in which within-class divergences are made explicit, the integration of determinants usually reviewed separately, and an explicit statement of the evidence level behind every entry. We examine food effects, the interaction between acid-suppressive therapy and pH-dependent agents, the dominant role of cytochrome P450 3A4 and of the efflux transporters P-glycoprotein and breast cancer resistance protein, and the exposure–response relationships that motivate therapeutic drug monitoring, for which the evidence remains uneven and does not yet support routine use. We propose a structured framework for operationalizing these principles in daily practice.

Pharmaceutics doi: 10.3390/pharmaceutics18091081

Authors: Na-Young Yu Kyu-Jin Cho Saeeun Ryu Gyulim Kim Jae-Woo Shin Donghee Park Jongho Won Young-Kee Shin Eun-Ah Kim Sang-Goo Cho Nae-Won Kang Byung-Hoon Lee Nam-Young Kim Eun-Seong Kim Dae-Duk Kim

Background/Objectives: Androgenetic alopecia (AGA) is the most prevalent form of hair loss. Although topical minoxidil (MNX) is widely used to treat AGA, its efficacy is limited by poor penetration across the stratum corneum. This study evaluated radiofrequency (RF) microporation as a means of enhancing cutaneous MNX delivery and hair-regrowth efficacy in a dihydrotestosterone (DHT)-induced AGA mouse model. Methods: RF-induced skin permeabilization and barrier recovery were assessed in rats using methylene blue and rhodamine B staining. In vivo skin deposition and pharmacokinetic studies were conducted to quantify cutaneous MNX accumulation and systemic exposure. Hair-regrowth efficacy was evaluated in DHT-treated mice. Results: RF microporation generated transient microchannels in the stratum corneum, increased rhodamine B penetration into deeper skin layers, and allowed substantial barrier recovery within 24 h. RF pretreatment significantly increased MNX deposition in the epidermis/dermis by 2.40-fold at 1 h and 1.94-fold at 3 h compared with topical MNX alone. RF-assisted topical administration resulted in a relative bioavailability of 11.17%, compared with 4.74% for topical administration without RF, while dose-normalized systemic exposure remained substantially lower than that following oral administration. In the AGA model, RF-assisted MNX treatment significantly increased hair coverage, length, and shaft thickness. Histological analysis further showed more prominent follicular structures in the RF-assisted MNX groups. Conclusions: RF microporation creates transient microchannels that enhance cutaneous MNX delivery and improve hair-regrowth efficacy. Importantly, RF-assisted topical administration maintained substantially lower systemic exposure than oral administration, supporting its potential as a needle-free strategy for topical AGA therapy and further translational evaluation.

Pharmaceutics doi: 10.3390/pharmaceutics18091080

Authors: Justyna Srebro Witold Brniak Paulina Poloczek Marian Paluch Aleksander Mendyk

Background/Objectives: Bisoprolol fumarate (BF) is a cardioselective beta-blocker used to treat pediatric heart failure and certain tachyarrhythmias. Currently, it is administered mainly as extemporaneously prepared suspensions from crushed tablets, which may cause issues with dose accuracy, stability, and palatability. Orodispersible minitablets (MODTs) may facilitate administration and individualized dosing, while prolonged release could potentially reduce peak-to-trough fluctuations and, thus, minimize adverse effects such as hypotension or bradycardia. This proof-of-concept study aimed to develop spray-dried prolonged-release microparticles containing BF, incorporate them into MODTs, and evaluate the effect of compression on drug release. Methods: Microparticles were prepared by spray drying ethanolic solutions of BF and ethylcellulose at 40 °C, 50 °C, and 60 °C. Their characterization included scanning electron microscopy, X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis and dissolution studies. Selected microparticles were compressed into 3 mm MODTs, which were evaluated for mechanical properties, disintegration, and BF release. Results: Formulations containing 10% BF and 90% ethylcellulose released from 52.6% to 73% of BF after 2 h, increasing to 84.8–90.0% after 8 h and reaching 96% after 24 h. Solid-state analyses indicated complete amorphization of the drug in these microparticles. The spray-drying temperature affected process efficiency but did not significantly impacted morphology or dissolution behavior. The optimized MODTs disintegrated in less than 30 s and had a tensile strength of up to 2.62 MPa. Importantly, compression increased the initial BF release from 26.4% for the microparticles to 43.9% for the MODTs at 0.5 h, resulting in f2 values of 45.04–48.46. Despite the higher initial BF release, the moderately prolonged-release profile was maintained in the case of MODTs. Conclusions: These findings demonstrate the feasibility of combining prolonged-release microparticles with orodispersible minitablets as a proof-of-concept for a pediatric drug delivery approach.

Pharmaceutics doi: 10.3390/pharmaceutics18091079

Authors: Mark Mandrik Veronika Makarova Ludmila Korol Ivan Sadkovskii Ivan Krasnyuk Sergey Antonov

Background: Hot-melt extrusion (HME) is a scalable pharmaceutical technology increasingly relevant to flexible manufacturing, including small-batch production, personalized dosage-form development, and potential use in pharmacy compounding. When translated into compounding practice, however, HME introduces a risk that routine quality-control methods available in pharmacies may be insufficient to reliably assess the stability of extrusion-based preparations. Methods: Granules containing 50% (w/w) darunavir were prepared by HME at 70 and 90 °C using a previously developed polymeric premix. Samples were stored for 24 months under ambient conditions. During storage, routine quality attributes were evaluated, including appearance, particle size distribution, loss on drying, disintegration time, content uniformity, and assay. Solid-state changes were investigated using differential scanning calorimetry (DSC) and X-ray diffraction (XRD), with a reference PEG-associated darunavir sample prepared and characterized for comparative analysis. Changes in drug release and darunavir content were assessed by dissolution testing and HPLC analysis, respectively. Results: Granules produced at both extrusion temperatures retained acceptable routine quality attributes throughout the 24-month storage period. No substantial changes were detected by visual inspection, pharmacopoeial tests, or UV assay. However, DSC revealed a new thermal event after storage, while XRD showed the formation of a new crystalline phase. Comparison with the reference PEG-associated sample supported the assignment of this phase as a PEG-associated crystalline phase of darunavir. Importantly, this transformation occurred even though the routine quality attributes evaluated in pharmacy compounding practice remained unchanged. Dissolution profiles differed between samples tested immediately after preparation and after long-term storage, with a more pronounced overall difference for granules produced at 90 °C, whereas HPLC confirmed comparable darunavir content in all investigated samples. Discussion: Our results show that routine compounding quality control can meet conventional acceptance criteria while failing to detect API solid-state changes in the investigated HME-derived system. In the PEG-containing matrix, amorphous darunavir undergoes storage-induced crystallization, forming a PEG-associated crystalline phase consistent with its known affinity for polyol-containing media. Conclusions: Acceptable routine quality attributes do not necessarily reflect the solid-state stability of APIs in HME-based formulations. These results highlight the need for solid-state risk assessment when developing extrusion-based systems intended for pharmacy compounding and other personalized manufacturing models in which routine quality control may not include advanced solid-state characterization.

Pharmaceutics doi: 10.3390/pharmaceutics18091078

Authors: Mantosh Kumar Satapathy Ting-Lin Yen Jing-Shiun Jan Ruei-Dun Tang Jia-Yi Wang Rajeev Taliyan Chih-Hao Yang

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Pharmaceutics doi: 10.3390/pharmaceutics18091077

Authors: Diana-Maria Mateescu Dragos-Mihai Gavrilescu Ruxandra-Ioana Mincioaga Ana-Maria Pah Ana-Olivia Toma Daniela-Vasilica Serban Cristiana Adina Avram Maria-Laura Craciun Bogdan Enache Camelia-Oana Muresan

Background/Objectives: Glycyl-L-histidyl-L-lysine (GHK) and its copper(II) complex GHK-Cu have been studied for matrix remodeling, inflammation, redox regulation, angiogenesis, and tissue repair, yet the literature frequently treats GHK-Cu as a single active ingredient despite formulation-dependent variation in coordination state, speciation, stability, pharmacokinetics, and toxicity. We critically evaluate GHK-Cu simultaneously as a bioactive metallopeptide and as a drug-delivery cargo, with explicit separation of apo-GHK, canonical GHK-Cu, GHK-derived copper peptides, and non-GHK copper-peptide systems. Methods: We conducted a structured systematic evidence-mapping review of PubMed/MEDLINE, Europe PMC, major publisher platforms, ClinicalTrials.gov, backward citation chains, and official European Union, U.S., and ICH regulatory sources from database inception through 12 August 2026. Biological evidence level and chemical/formulation quality were graded independently using an author-defined two-axis framework. Delivery studies were extracted against a fixed matrix comprising formulation, claimed loading, molar copper occupancy, labile copper, species-resolved release, factorial controls, stability/manufacturability, and objective outcome. Quantitative pooling was not performed because active-entity definitions, formulations, doses, models, comparators, and endpoints were not quantitatively commensurable. Results: Preclinical data consistently support effects on matrix remodeling, epithelial repair, inflammatory/redox regulation, and angiogenesis, but the clinical evidence remains sparse and does not meet contemporary active-entity quality standards. Historical cosmetic reports are small or incompletely characterized; a 13-participant post-CO2-laser study was negative on objective endpoints, whereas a 2026 18-participant split-face eyebrow study reported positive cosmetic hair outcomes but did not define GHK-Cu speciation or local exposure. The ongoing phase 2 acute-wound study NCT07437586 remains recruiting and has no efficacy results; its registration cannot be used as evidence of clinical translation. Across delivery studies, particle size, polydispersity, encapsulation efficiency, total peptide/copper content, and bulk release are often reported, whereas molar occupancy, labile copper, and release of intact GHK-Cu versus apo-GHK/free copper are usually not resolved. Conclusions: Translation is limited less by biological plausibility than by pharmaceutical definition and evidence attribution. We define the “active pharmaceutical entity” operationally as the reproducible chemical state intended to mediate pharmacology at administration, not as an established regulatory designation or a claim that one immutable molecular species persists in biological fluids. We further propose, explicitly as an author-derived development framework rather than a consensus standard, a control strategy based on molar occupancy, route-specific labile copper specifications, orthogonal speciation, mechanism-linked potency, species-resolved release, factorial controls, route-specific safety decision thresholds, ICH-aligned stability, and GMP-scalable manufacture. Until these requirements are met and controlled clinical efficacy is demonstrated, GHK-Cu should be regarded as a promising but unproven therapeutic cargo rather than a clinically validated regenerative drug.

Pharmaceutics doi: 10.3390/pharmaceutics18091076

Authors: Agata Olszewska Maria Brycka Anita Umerska Lidia Tajber Marek Pyda Marcin Skotnicki

Objective: In this study, the solid-state properties of two amorphous forms of rosuvastatin calcium (ROS) and co-amorphous systems of rosuvastatin with carvedilol (CAR) were investigated. Methods: The studied systems were investigated using TGA, DSC, TMDSC, PXRD, SAXS, SSNMR, and DVS. Results: DSC, TMDSC and SAXS revealed subtle differences between as-received ROS (ROS AR) and freshly prepared amorphous ROS (ROS AM), providing evidence of relaxation polyamorphism. Analysis of the scanning-rate dependence of the glass transition temperature classified ROS as a fragile glass former. Water sorption studies of ROS AR using Brunauer–Emmett–Teller analysis, the Guggenheim–Anderson–de Boer equation, and Young–Nelson models indicated a moderate specific surface area and multilayer adsorption, with water binding both at the surface and within the bulk. Co-amorphous CAR–ROS systems in various molar ratios exhibited single glass transitions, confirming the formation of homogenous amorphous phases. Negative deviations from the Couchman–Karasz equation indicated non-ideal mixing behaviour. Physical ageing studies showed high stability of the ROS AM and CAR-ROS 1:1 co-amorphous system, which remained amorphous after three years of storage. Conclusions: Overall, ROS forms stable amorphous and co-amorphous systems with CAR, exhibiting favourable miscibility, structural stability, and potential for application in fixed-dose combination formulations.

Pharmaceutics doi: 10.3390/pharmaceutics18091075

Authors: Nader Amanatchi Ilyesse Bihi Matthieu Briet Wim De Malsche Karine H. Hellemans

Background/Objectives: Poly(lactic-co-glycolic acid) (PLGA) microparticles are used for sustained drug delivery, yet their final quality depends on both initial droplet formation and the subsequent solvent-extraction-driven droplet-to-particle transition. Microfluidics provides excellent control over precursor droplets, but this control may be partially lost during particle formation. We previously developed dynamic solvent extraction (DSE), in which the droplet-to-particle transition occurs progressively during continuous transport through an extended microfluidic channel. Here, we investigated whether this control improves quality attributes and release of drug-loaded PLGA microparticles. Methods: Dexamethasone (DEX)-loaded PLGA microparticles were produced using DSE or static solvent extraction (SSE), in which microfluidic droplets were transferred to an external aqueous medium for particle formation. Three PLGA concentrations (2.5, 5, and 10% (w/v)) were investigated, with processing conditions selected to obtain comparable final particle sizes and reduce size as a confounding variable. Results: DSE significantly reduced particle-size coefficient of variation compared with SSE, indicating better preservation of size uniformity. Drug loading and encapsulation efficiency were governed mainly by PLGA concentration, with no significant effect of extraction strategy. Morphological effects were formulation-dependent, with the most pronounced defects in 5% (w/v) SSE particles. All formulations provided sustained DEX release over 70 days. Although the overall effect of extraction strategy on 24 h burst release was not statistically significant, DSE consistently produced numerically lower burst release, with the largest and most variable burst observed for the morphologically heterogeneous 5% (w/v) SSE formulation. Conclusions: These findings extend DSE to drug-loaded sustained-release microparticles and identify the solvent-extraction environment as an important determinant of final particle quality.

Pharmaceutics doi: 10.3390/pharmaceutics18091074

Authors: Maria Medina de Azevedo Maria Luiza Fidelis da Silva Gabriela Pereira da Silva Joyner David Anaya Miranda Annye Vitória Moraes Luana Ale Bertoncello Pael Telma Lélia Gonçalves Schultz de Carvalho Giselle Nathaly Calaça Thainá Aparecida Rafael Silva Bianca Viana Silva Thaylla Bianca de Almeida Vilela João Pedro Vilella Neto Ariany Carvalho dos Santos Ana Paula de Carlos Sela Monique Assis de Vasconcelos Barros Francislaine Aparecida dos Reis Lívero Otávio Akira Sakai Érica Marusa Pergo Coelho Arquimedes Gasparotto Junior

Background: Hypertensive disorders of pregnancy present a major global health challenge. Green nanotechnology offers a strategy to enhance the stability and efficacy of bioactive natural products. This study evaluated gold nanoparticles green-synthesized with Pfaffia glomerata extract (AuNPs-PG) against gestational hypertension and fetal complications in spontaneously hypertensive rats (SHRs). Methods: P. glomerata root extract was characterized by UHPLC-MS/MS. AuNPs-PG were green-synthesized with 2 mmol/L HAuCl4 (1:5 v/v) at 40 °C and characterized by UV-Vis and DLS. Pregnant Wistar-Kyoto and SHRs were divided into naive, negative control (NC), amlodipine (5 mg/kg), and AuNPs-PG groups (0.03, 0.1, and 0.3 mg/kg) treated daily on gestational days 1–18 to evaluate maternal cardiovascular, renal, biochemical, and reproductive outcomes. Results: UHPLC-MS/MS identified 38 compounds. AuNPs-PG showed a plasmon band at 520–550 nm and 88.1 nm hydrodynamic diameter. In SHRs, 0.3 mg/kg AuNPs-PG reduced systolic blood pressure by 18.5% and mean arterial pressure by 15.4%, while mitigating vascular dysfunction by lowering phenylephrine vasoconstriction by 52.7% and boosting acetylcholine vasodilation by 239.3%. Treatment shortened QTc interval by 26.4%, normalized T-wave inversion, and restored renal excretion, increasing urinary volume by 75.0% and sodium by 94.2%. Furthermore, AuNPs-PG elevated placental growth factor by 180.9%, lowered malondialdehyde by 63.5%, and preserved placental histology. Consequently, mean fetal weight increased by 21.6%, implantation index by 50.5%, and offspring-to-mother ratio by 129.6%. Conclusions: AuNPs-PG (0.3 mg/kg) effectively mitigate maternal and fetal complications in gestational hypertension. Gold nanoparticles serve as nanocarriers, whereas surface-adsorbed phytochemicals drive the biological effects.

Pharmaceutics doi: 10.3390/pharmaceutics18091073

Authors: Sophia Leslie Stella Rios Hana Elnahas Megan Keniry

Clinical outcomes for brain cancer are often poor because the blood–brain/tumor barrier hinders effective drug delivery to malignant tissue. Glioblastoma, the most common primary brain malignancy in adults, has an average survival of approximately fourteen months. Here, we discuss novel strategies that our research group and others are developing to deliver chemotherapy to the brain via the nasal cavity. Although significant hurdles remain, intranasal delivery holds substantial promise for improving outcomes for patients with brain cancer. Intranasal delivery is noninvasive, permits repeated dosing, and has been shown to enable direct nose-to-brain transport that bypasses the blood–brain barrier. Challenges such as accurately targeting drugs to the appropriate region of the nasal cavity at therapeutically relevant doses, while maintaining reproducibility, make this cutting-edge approach a regulatory challenge. The prolonged path to clinical translation discourages many researchers from pursuing this potentially life-saving strategy. Nevertheless, preclinical studies demonstrate that intranasal delivery can achieve up to ten-fold higher concentrations of select drugs in the brain. Cancer chemotherapeutics span a wide range of molecular formats, from small molecules to 150-kilodalton antibodies. Accordingly, delivery strategies must be carefully matched to the molecular properties of each therapeutic. Here, we focus on the intranasal delivery of small molecule inhibitors using nanomaterial-based platforms, including aerosols, lipids, gold nanoparticles, gels, emulsions, fibers, and their combinations. Ultimately, we hope that intranasal delivery approaches will be translated to provide patients with better therapeutic outcomes.

Pharmaceutics doi: 10.3390/pharmaceutics18091072

Authors: Nur Aisyah Ikra Nurohman Sriwidodo Sriwidodo Patihul Husni Cecep Suhandi Gofarana Wilar Ahmad Choibar Tridakusumah Sabreena Safuan

Background/Objectives: Mangosteen pericarp extract (Garcinia mangostana L.) exhibits robust antioxidant properties. However, its pharmaceutical application is hindered by poor water solubility and low physicochemical stability. This study aimed to develop a lipid-based nanoemulsion to overcome these limitations and assess its physicochemical characteristics, radical scavenging capacity, and dissolution profiles. Methods: Nanoemulsions were prepared by high-shear homogenization followed by ultrasonication method using sacha inchi oil (Plukenetia volubilis L.) across varying hydrophilic–lipophilic balance (HLB) values. Formulations were characterized by emulsion type, pH, rheology, droplet size, surface charge, encapsulation efficiency, and morphology via transmission electron microscopy. Antioxidant capacity was quantified using the DPPH assay, while in vitro dissolution experiments measured solubility enhancement relative to unformulated extract. Results: All formulations formed stable oil-in-water systems without phase separation. The HLB 10 formulation exhibited optimal performance, yielding a mean droplet diameter of 490.03 ± 32.12 nm, zeta potential of −52.73 ± 0.09 mV, and encapsulation efficiency of 89.14 ± 0.19%. Micrographs revealed well-structured spherical droplets. The raw extract showed strong antioxidant activity (IC50 = 21.68 ppm), which remained functional in nanoemulsion (IC50 = 84.48 ppm). Dissolution testing indicated a 13.4-fold improvement over raw powder. However, storage evaluation indicated chemical loss of bioactive constituents over one month. Conclusions: Sacha inchi oil-based nanoemulsions significantly enhance the dissolution rate of mangosteen pericarp extract while maintaining functional antioxidant potential. Although chemical stability during storage remains a limitation requiring further optimization, this nanocarrier platform offers strong potential to overcome solubility barriers for oral bioactive delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18091070

Authors: Huaying Pan Juan Yang Jiaqi Fu Shenao Yan Yue Yan Yining Xu Minglu Zhou Lian Li Yucheng Xiang

Nanodrug delivery systems have shown great promise against tumors, inflammatory diseases and central nervous system disorders. However, several limitations still exist before further clinical application. The unsatisfactory circulation time in vivo, rapid clearance by the mononuclear phagocyte system, insufficient tissue penetration and low targeting efficiency all hinder the clinical translation of nanomedicines. Cells are the fundamental functional units of the body and their intrinsic biological properties make them promising drug delivery vehicles for targeted therapy. Conjugating cells with nanomedicines combines the carrier functions of living cells with the therapeutic effects of nanodrugs, offering an effective strategy to improve targeted drug delivery. Compared with the existing literature, this review specifically summarizes the biological properties and major applications of six types of living cell carriers. The recent strategies for constructing cell–nanomedicine conjugates are also discussed. Finally, we highlight the current challenges for clinical translation and discuss future directions for the development of cell–nanomedicine delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18091071

Authors: Sandra Miočić Andrea Rašić Jelena Torić Michela Abrami Kristina Ferderber Biserka Cetina-Čižmek Mario Grassi Jelena Filipović-Grčić

Objectives: The physical stability of concentrated azelaic acid nanosuspensions cannot be predicted solely from interfacial stabilisation but reflects the interplay between particle size-dependent thermodynamic driving forces, polymer-mediated rheological structuring and the resulting bulk microstructural organisation. This study employed an A-optimal design of experiments (18 runs) to investigate formulation–process relationships in concentrated azelaic acid nanosuspensions (10–20% w/w) stabilised with a dual HPMC–chitosan system. Methods: Particle size, ζ-potential, rheological behaviour, solid-state properties and in vitro permeation across Strat-M® membranes were evaluated, and formulation–process relationships were analysed using multivariate modelling and logistic regression. Results: Particle size (342–1118 nm; R2 = 0.97) was primarily governed by the applied milling regime. Logistic regression demonstrated a size-dependent probability of crystal growth, with the estimated particle-size transition point (predicted probability = 0.5) shifting from approximately 424 nm after preparation to 594 nm following accelerated storage at 40 °C. CHI concentration controlled ζ-potential (+18.5 to +47.9 mV), although ζ-potential alone did not adequately explain the observed storage stability. Where measurable, zero-shear viscosity (68–45,462 mPa·s) reflected substantial differences in low-shear rheological structuring, while formulations containing higher HPMC concentrations generally exhibited improved stability, consistent with polymer-mediated kinetic constraints on crystal growth. In vitro permeation studies using Strat-M® membranes demonstrated permeation behaviour consistent with structured diffusion-controlled systems, exhibiting lower flux but more uniform permeation profiles than the reference formulation under the applied experimental conditions. Conclusions: Integration of multivariate modelling with rheological, solid-state and permeation characterisation provided an integrated understanding of the formulation–process relationships governing the short-term physical stability and comparative in vitro transport behaviour of concentrated dermal AZA nanosuspensions.

Pharmaceutics doi: 10.3390/pharmaceutics18091069

Authors: Binaya Sapkota Susmita Phuyal Arjun Dhwoj Bamjan Seung-Sik Cho Jung-Hyun Shim Jin Woo Park Laxman Subedi

Topical and transdermal dosage forms can localize therapy or provide controlled systemic exposure, but translation is limited by the selective stratum corneum (SC) barrier and post-application changes in formulation. Existing reviews often address individual enhancers or carriers without integrating drug properties, vehicle microstructure, post-application transformation, quality of evidence, and product development constraints. This review fills that gap by comparing passive formulations, chemical permeation enhancers, vesicular and lipid-based carriers, supersaturating systems, and physical barrier bypass technologies within a skin barrier-informed framework. Increased permeation alone does not establish translational value. Passive delivery remains most feasible for potent, moderately lipophilic small molecules. Chemical enhancers are scalable but limited by irritation and drug-dependent compatibility; nanocarriers may improve solubilization and cutaneous deposition but often lack human confirmation; and physical methods broaden delivery to macromolecules while adding device, manufacturing, usability, and regulatory burdens. Solvent evaporation, residual film composition, supersaturation, precipitation, and drug–vehicle affinity further determine the effective post-application driving force. Accordingly, we propose an evidence-ranked framework linking payload properties and target compartment to mechanism, safety, clinical readiness, and regulatory complexity. It distinguishes mechanistic promise from clinically demonstrated delivery and identifies the evidence needed to advance reproducible topical and transdermal products.

Pharmaceutics doi: 10.3390/pharmaceutics18091068

Authors: Rumeysa Ceylan Eren Aytekin Heybet Kerem Polat Ozan Kaplan Mustafa Çelebier Sibel Bozdağ Pehlivan

Background/Objectives: The aim of this study was to develop and validate a simple, accurate, reproducible, and sensitive reverse-phase high-performance liquid chromatography method for the quantification of reproxalap (RP) in cyclodextrin (CD) inclusion complexes and poloxamer 407 hydrogel formulations. Methods: Chromatographic separation was achieved using a mobile phase of deionized water and an organic phase (methanol:acetonitrile, 55:45 v/v) in a 40:60 (v/v) ratio, at a flow rate of 0.9 mL/min and a run time of 10 min. Validation assessed linearity, specificity, accuracy, and sensitivity over a concentration range of 1–60 μg/mL. Phase-solubility studies were conducted to determine the apparent stability constant (K1:1), and inclusion complexes were characterized by Fourier transform infrared spectroscopy and differential scanning calorimetry. Results: The method demonstrated specificity, linearity, sensitivity, and accuracy within the tested concentration range of 1–60 μg/mL. The apparent stability constants (K1:1), calculated from the phase-solubility plot slopes, were 2666.66 M−1 for Sulfobutyl Ether β-Cyclodextrin (SBE-β-CD) and 526.3 M−1 for Hydroxypropyl-β-Cyclodextrin (HP-β-CD). The solubility of RP in deionized water (0.25 mM) increased 8.6-fold to 2.15 mM with the HP-β-CD inclusion complex and 22.4-fold to 5.6 mM with the SBE-β-CD inclusion complex. The method was successfully applied to determine RP in inclusion complexes and poloxamer 407 hydrogel formulations, with no interference from formulation excipients. Conclusions: The validated RP-HPLC method is suitable for quality control analysis of RP in CD inclusion complexes and poloxamer 407 hydrogel formulations, supporting its application in the future development of these drug delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18091067

Authors: Lin Zhang Wenbo Yang Shipu Jia Jing Shang Jincheng Wang Xin Zhao Haotian Bai Chenyu Wang

Objectives: Critical-sized bone defects continue to represent a substantial challenge in orthopedic clinical practice. Decellularized bone matrix (DBM) possesses favorable osteoconductive properties due to its retention of native extracellular matrix architecture and collagen components. However, its limited osteogenic bioactivity restricts its application in complex bone defect repair. This study aimed to construct a dendritic mesoporous silica (DMSN)-modified DBM composite scaffold loaded with teriparatide (DBM-DMSN@TPTD) and to systematically evaluate its physicochemical properties, drug release behavior, biocompatibility, and osteogenic differentiation-promoting capacity. Methods: A DBM scaffold was prepared from bovine femoral cancellous bone via a combined freeze–thaw and chemical detergent decellularization method. DMSNs were synthesized through a sol–gel method, amine-functionalized with APTES, and covalently grafted onto the DBM surface via EDC/NHS crosslinking. Teriparatide was loaded onto the composite scaffolds at three concentrations (1, 10, and 100 nmol/L). The scaffolds were characterized via SEM, TEM, BET, EDS and XPS. Decellularization efficacy was assessed by DAPI staining and nucleic acid quantification. Drug release behavior was evaluated through in vitro release studies, while biocompatibility and osteogenic differentiation of rat BMSCs were examined using Live/Dead staining, phalloidin/DAPI cytoskeletal staining, CCK-8 assays, ALP staining, and RUNX2/OCN immunofluorescence. Results: DMSNs demonstrated a dendritic mesoporous architecture, featuring a specific surface area of 390.44 ± 1.78 m2/g and pore diameters within the range of 15–20 nm. DBM showed effective removal of immunogenicity, with well-preserved collagen architecture. Drug release displayed a biphasic pattern, with 56.03% released within the first 72 h and 83.23% by day 16. None of the tested scaffolds showed obvious cytotoxicity under the experimental conditions. The DBM-DMSN@TPTD-M group (10 nmol/L) produced the strongest effects on BMSC proliferation and osteogenic differentiation, as indicated by the highest ALP activity and elevated RUNX2 and OCN expression (p < 0.05). Conclusions: The DBM-DMSN@TPTD scaffold offers a native bone microenvironment, sustained drug release, and osteogenic activity in vitro. These features may support BMSC proliferation and osteogenic differentiation. Accordingly, this scaffold warrants further investigation as a potential strategy for bone defect repair.

Pharmaceutics doi: 10.3390/pharmaceutics18091066

Authors: In Gyu Yang Jun-Young Han Min Young Jeong Dong-Wan Seo Myung Joo Kang Sun Ho Kim

Objectives: Fixed-dose combination tablets containing sitagliptin hydrochloride (SG) and metformin hydrochloride (MF) are a mainstay in the clinical management of type 2 diabetes. However, SG is highly susceptible to chemical degradation during storage, particularly in the presence of MF. Herein, a bilayer tablet in which SG and MF are physically separated into distinct layers was designed to enhance the chemical stability of SG while ensuring pharmacokinetic equivalence to the marketed reference product Janumet®. Methods: The compositions of individual SG and MF compartments were selected based on evaluations of their physical properties and dissolution profiles. Critical process parameters, including the pre- and main compression forces and coating levels of bilayer tablets, were fine-tuned to achieve dissolution characteristics comparable to those of the reference product. Results: Under accelerated storage conditions (40 °C, 75% relative humidity), the optimized bilayer tablet exhibited superior stability, with total SG-related impurity levels of 0.18% compared with 0.86% in the reference product after six months. Furthermore, in a randomized bioequivalence study in healthy volunteers (n = 30), the SG/MF bilayer tablet was pharmacokinetically equivalent to the reference product, with all parameters falling within the Food and Drug Administration-mandated regulatory criteria. Conclusions: In conclusion, this SG/MF bilayer tablet has better storage stability than the reference product and may be an alternative to conventional SG/MF combination tablets.

Pharmaceutics doi: 10.3390/pharmaceutics18091065

Authors: Moataz M. Rashad Shaimaa M. Kasem Khaled E. El-Kelany Sara A. Abdel Gaber

Purpose: Cryptosporidiosis is a parasitic inflammatory disease that causes the death of around 1.6 million people annually worldwide and is associated with Cryptosporidium parvum (C. parvum) infection. This study aimed to evaluate the therapeutic efficacy of carvacrol (CV) loaded into mesoporous silicate nanoparticles (MSNs) against C. parvum using in vitro, in vivo, histopathological, immunohistochemical, biochemical, and computational approaches. Methods: In vitro assays were conducted to evaluate the oocysticidal activity of CV-loaded MSNs (CV-MSNs) for 96 h. In silico assays were carried out to investigate target inflammatory proteins involved in cryptosporidiosis. In vivo tests were performed on male Swiss albino mice (22 ± 5 g, 4–6 weeks) to assess the anti-inflammatory and antioxidant effects of CV after loading onto MSNs. Results: CV showed its highest oocysticidal efficacy after loading onto spherical 72 nm MSNs (MSN2), exhibiting a 0.07 mg/mL LC50. At the in vivo level, CV-MSN2 markedly restored ileal, pulmonary, and hepatic histoarchitecture and normalized biochemical indices in infected mice. CV-MSN2 also showed the strongest inhibitory action against computationally selected target proteins, TLR-4, NF-κb-P65, STAT-3, NOS2, and JAK-1, with accompanying downregulation of IL-6, IL-1β, and TNF-α. Similarly, antioxidant markers were significantly decreased following CV-MSN2 treatment compared to the positive controls. Conclusions: This study shows that MSNs enhanced CV efficacy against ileal inflammation induced by C. parvum, preserving CV’s molecular targets.

Pharmaceutics doi: 10.3390/pharmaceutics18091064

Authors: Noelia Fernández-Ceballos Laura Gallego-Yerga Rafael Peláez

Background: Antimitotic agents are very successful antitumor therapies, but lack tumor selectivity, causing toxicity. Antitumor alkylphospholipids (APLs) selectively accumulate in tumor cells but display low potencies. Hypothesis: Incorporating APL moieties onto antimitotic N-trimethoxyphenyl naphthalene sulfonamides (TMNS) might afford SMDCs with the potency of antimitotics and the tumor selectivity of APLs. Methods: 24 new TMNSs with spacers of different lengths (4 to 9 atoms) and nature (alkanes or ethers) on the sulfonamide nitrogen and capped with phosphorus-containing groups such as diethylphosphonates, phosphonic acids, and hydrogenophosphonate esters of aminoalcohols (diethylaminopropanol, choline, or dimethylaminoethanol) were designed and synthesized. Their antiproliferative effects against several cancer cell lines and their cotreatment with verapamil to assess whether they are substrates of MDR pumps were evaluated. The mechanism of action was studied: cell cycle effects, apoptosis induction, and immunofluorescence microscopy. Computational studies considered binding to tubulin and pharmacokinetics. Results: Diethyl phosphonates and phosphonic acids are antiproliferative in the micromolar to submicromolar range. P-gp inhibitor verapamil renders inactive compounds active, suggesting that efflux, not binding, removes activity. Mechanistic studies agree with an antimitotic action. Proposed binding to tubulin is similar to TMNS, with the phospholipid-like substituent projecting towards the interdimer space. Conclusions: Hybridization of TMNSs with potentially tumor–targeting APLs yields microtubule-disrupting antitumor compounds. However, the modifications assayed turn the compounds into substrates of MDR. These compounds are a proof of concept of the strategy that might succeed if future modifications avoid MDR and might target the compounds towards cancer cells.

Pharmaceutics doi: 10.3390/pharmaceutics18091063

Authors: Saif-alnasr H. Mohammed Ayman B. Mousa Mohammed Taj-Eldin Abdalla Anita Sztojkov-Ivanov Kálmán F. Szűcs Róbert Gáspár

Background: Sex hormones play crucial functions in the body via the genomic and non-genomic pathways. 5α- and 5β-dihydrotestosterone (5α- and 5β-DHT) are 5-reduced testosterone metabolites. We aimed to investigate the non-genomic effect of 5α- and 5β-DHT on uterine muscle contractility in vitro and in vivo for non-pregnant and 22-day-pregnant rats. Methods: The rapid in vitro action of 5α-DHT and 5β-DHT (10−9–10−3 M) on KCl (25 mM)-stimulated contractions was examined in an organ bath in the presence of several blockers and after endometrium removal. The actions of DHTs (10−4 M) and nifedipine (10−7 M) were also examined in contractions stimulated by KCl (40 mM) with a cumulative addition of CaCl2 (3–120 mM). Plasma DHT levels were measured by ELISA after a single intraperitoneal (i.p.) administration of DHT (10 mg/kg), and kinetic curves were obtained. The in vivo relaxing action of DHTs was detected by strain-gauge sensors. The animals received 5α- or 5β-DHT alone (3/10/30/100/300 mg/kg i.p.) or with flutamide (100 mg/kg i.p.). Results: DHT showed concentration-dependent relaxation of uterine muscle in vitro, with a higher potency observed for 5β-DHT. Among the blockers used, G15 and L-NAME reduced the potency of 5α-DHT in pregnant rats only. Both DHTs inhibited the contraction-increasing effect of CaCl2, proving their Ca2+ inhibiting effects. DHTs had similar cmax and tmax values in both non-pregnant and pregnant rats. DHT plasma levels before and 30 min after administration were proportional to the administered doses. Their single doses (30/100/300 mg/kg) elicited a flutamide-resistant uterine relaxing effect in vivo. Conclusions: DHTs or their analogs are candidates for further studies in the human uterus to establish their potential to treat conditions associated with uterine hyperactivity.