Pharmaceutics doi: 10.3390/pharmaceutics18070778

Authors: Liqin Huang Yao Fu Fang Li

Keratoconus, a progressive corneal ectasia, remains a major cause of irreversible visual impairment worldwide. Conventional corneal cross-linking (CXL) with riboflavin/ultraviolet A (UVA) has revolutionized clinical management, yet its efficacy is still constrained by epithelial barriers, oxygen dependence, and safety concerns in thin corneas. Emerging nanotechnology provides a transformative opportunity to overcome these bottlenecks. This review highlights the enhancement of riboflavin delivery efficiency by nanocarriers, the photodynamic optimization of nano-enhanced cross-linking agents, and the synergistic strengthening effect of nanocomposites on corneal mechanical strength. We emphasize not only their potential to enhance drug penetration, improve cross-linking efficiency, and extend clinical indications, but also their role in advancing toward a new generation of personalized, intelligent, and minimally invasive corneal therapy. Finally, we discuss translational challenges, including manufacturing, long-term biosafety, and regulatory frameworks, and present a theoretical roadmap that integrates nanotechnology, real-time imaging, and artificial intelligence (AI)-assisted decision-making to achieve a closed-loop “sense–decide–act” therapeutic system. By situating nanomaterial-enhanced CXL within precision ophthalmology, this review highlights its capacity to redefine the standard of care for keratoconus and related ectatic disorders.

Pharmaceutics doi: 10.3390/pharmaceutics18070777

Authors: Qian Zhang Yuhang Li Jiahui Zhang Xuewen Zhao Danlu Li Wenting Zhao Xin Hai Xin Chen Xinlei Yang Jingxin Gou Chunpeng Zhang Xing Tang Yilei Zhao

Background/Objectives: Glioblastoma multiforme (GBM) treatment is limited by tumor hypoxia and poor specificity of therapeutic agents. To address these challenges, we developed brain-targeted liposomes co-encapsulating 5-aminolevulinic acid (5-ALA) and catalase (CAT), termed brain-targeted 5-ALA-CAT liposomes (BACL), which were surface-modified with the Angiopep-2 ligand to enhance blood–brain barrier penetration and achieve multimodal therapy combining targeted delivery and oxygen generation. Methods: BACL was prepared and characterized. Tumor targeting was verified by flow cytometry and in vivo imaging. In vitro antitumor activity was evaluated by wound-healing assay, colony formation assay, live/dead staining, MTT assay, and Western blotting. In vivo efficacy, apoptosis, and safety were assessed in a subcutaneous xenograft model. Transcriptome sequencing and qRT-PCR were employed to identify molecular mechanisms and novel targets. Results: BACL exhibited favorable physicochemical properties (size: 122.4 nm, PDI: 0.189, zeta potential: −12.3 mV) and spherical morphology as observed by TEM, with encapsulation efficiencies of 51.2% for 5-ALA and 43.8% for CAT. Compared with unmodified 5-ALA, BACL increased the cellular uptake efficiency by 1.6-fold in glioma cells while maintaining catalytic stability for sustained oxygen generation. In vitro experiments demonstrated that BACL significantly inhibited glioma cell migration, colony formation, and cell viability, and induced apoptosis. In a subcutaneous xenograft tumor model, BACL-mediated photodynamic therapy (PDT) achieved a tumor growth inhibition rate of 52%, with apoptosis induction via regulation of Bcl-2, Bax, and p53 expression, and no obvious toxicity to major organs was observed. Transcriptomic analysis combined with qRT-PCR validation revealed that BACL activates multiple antitumor signaling pathways, including targeted inhibition of IL-10 and CXCL13 to disrupt cytokine–receptor interactions, as well as coordinated regulation of S100A3 and IGSF-9 expression to suppress glioma progression. Conclusions: These multimodal actions enhanced PDT efficacy while remodeling the tumor microenvironment. Our findings position BACL as a promising therapeutic platform integrating targeted delivery, hypoxia alleviation, and immunomodulation for GBM therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18070776

Authors: Yixuan Fang Jing Wu Shiyi Sun Yan Li Xingwu Ran

Diabetic foot ulcers (DFUs) are among the most severe complications affecting diabetic patients, and dressing therapy is one of the standard treatments for DFUs. However, traditional dressings are inadequate for addressing the complex microenvironment of DFUs. Consequently, advanced natural polymer-based dressings have attracted extensive research attention in diabetic foot care due to their biocompatibility, low immunogenicity, and biodegradability. These natural polymer materials include collagen, gelatin, chitosan (CS), hyaluronic acid (HA), alginate, and cellulose. This review systematically analyzes the pathophysiological mechanisms underlying the difficult healing of DFUs and the advantages of natural polymer-based dressings in diabetic wound healing, highlights preclinical studies, and synthesizes evidence from clinical research. Moreover, we pinpoint the challenges associated with these dressings and propose future directions for the improvement of diabetic wound care.

Pharmaceutics doi: 10.3390/pharmaceutics18070775

Authors: Rongrong Wang Yiying Zeng Zhaowu Zeng Tian Xie

Background/Objective: This study aimed to use multiple disk domain receptor 1 (DDR1), transforming growth factor β1 (TGFβ-1), tumor-associated calcium signal transduction protein 2 (TACSTD2), and polyligand proteoglycan 1 (SDC1) siRNA to treat pancreatic cancer with the goals of high specificity, significant therapeutic efficacy, and relatively low toxicity. Methods: (1) A microfluidic method was used to prepare siRNA-LNPs with different formulations. (2) Quantitative PCR (qPCR) and Western blot assays were used to detect the inhibitory effect of different-prescription siRNA-LNP formulations on mRNA and protein expression levels of related genes in PaTu 8988 pancreatic cells. (3) The anti-pancreatic cancer effect of multiple siRNAs combined with LNPs in vivo was evaluated using the BALB/c nude mouse model with subcutaneous pancreatic cancer xenografts. Results: (1) Three siRNA-LNP formulations, DMG, CE 1.5, and CE 0.75, were successfully prepared, exhibiting small particle sizes and uniform distribution. (2) qPCR and Western blot results indicated that DDR1, TGFβ-1, TACSTD2, and SDC1 siRNA-LNP significantly inhibited related genes’ mRNA and protein expression in pancreatic cancer PaTu 8988 cells. (3) Efficacy studies in animals indicated that multiple siRNA combined with LNPs in each group exhibited significant antitumor effects on pancreatic cancer tumor-bearing nude mice. The therapeutic efficacy of the combined siRNAs was superior to that of single siRNA treatments, indicating a clear combined effect, especially with three- and four-siRNA combinations. Conclusions: The prepared DDR1/TGFβ-1/TACSTD2/SDC1 siRNA-loaded LNP demonstrated a small particle size, high gene inhibition efficiency, and a significant therapeutic effect in treating pancreatic cancer. Its safety is generally acceptable, but attention should be paid to the toxicity caused by LNP excipients, especially cationic lipids.

Pharmaceutics doi: 10.3390/pharmaceutics18070774

Authors: Paola Franco Raffaella Belvedere Emanuela Pessolano Sara Liparoti Roberto Pantani Antonello Petrella Iolanda De Marco

In the original publication [...]

Pharmaceutics doi: 10.3390/pharmaceutics18070773

Authors: Federica Campana Emanuela De Bellis Floriana Porcaro Gaetano Di Guida Luigi Della Gravara Alberto Servetto Floriana Morgillo Federica Papaccio Amelia Filippelli Carminia Maria Della Della Corte Valeria Conti

For decades RAS mutations represented a chimera for drug developers, as all efforts put into attempting to pharmacologically inhibit them failed. Finally, in recent years, the advent of the KRAS G12C inhibitors sotorasib and adagrasib pushed the development of several new drugs; therefore, the landscape of RAS inhibitors evolved and now includes several compounds with different mechanisms of action. In this review we provide an updated summary of RAS inhibitor drugs, detailing their mechanism of action, pharmacokinetics and toxicity profile as well as efficacy studies.

Pharmaceutics doi: 10.3390/pharmaceutics18070772

Authors: Southamany Sisavengsouk Teeratas Kansom Boonnada Pamornpathomkul Porawan Aumklad Tanasait Ngawhirunpat Praneet Opanasopit Phuvamin Suriyaamporn

Background: Conventional transdermal drug delivery systems are often limited by poor skin permeability and low drug loading efficiency, necessitating the development of advanced delivery platforms. Objectives: This study aimed to develop and optimize photopolymerizable bioinks (PBs) for liquid crystal display (LCD)-based 3D printing of crosslinked hydrogel microneedles (cHMNs) to enhance transdermal delivery of estradiol valerate (E2V). Methods: A Box–Behnken design (BBD) was used to optimize the effects of Gantrez™ S-97, Jurymer™, and polyvinyl alcohol (PVA) on viscosity, exposure time, hardness, and elasticity, with strong predictive performance (R2 = 0.9702–0.9907). Results: Estradiol valerate-loaded nanoparticles (E2V-NPs) were prepared via ionotropic gelation, exhibiting a particle size of 698.33 (0.78) nm, PDI of 0.50 (0.06), zeta potential of −39.09 (7.32) mV, and high encapsulation efficiency (86.87 (0.78)%). The optimized PBs enabled fabrication of uniform cHMNs (~800 µm height) with adequate mechanical strength (hardness 20.45 (1.23) N; elasticity 2.97 (0.49) MPa) and effective insertion capability. The E2V-NPs-loaded cHMNs exhibited sustained drug release over 12 days (~56.92 (4.27)%). Skin permeation studies showed a significantly enhanced flux (10.81 (4.55) µg/cm2/h) and cumulative permeation (12.94 (2.06) µg/cm2) compared to topical E2V-NPs and suspension, along with increased skin accumulation (38.55 (0.10) µg). Cytotoxicity studies confirmed that E2V and E2V-NPs were biocompatible (>80% viability), while PBs showed concentration-dependent cytotoxicity. Conclusions: Overall, this integrated platform combining design of experiment, nanoparticles, microneedles, and LCD 3D printing offered a promising strategy for enhancing transdermal drug delivery efficiency and reproducibility.

Pharmaceutics doi: 10.3390/pharmaceutics18070771

Authors: Kinga Lis

Pantothenic acid (PA; vitamin B5) is an essential component of many key metabolic processes. Vitamin B5 deficiency causes dysfunction in various systems and organs. Humans do not produce vitamin B5, so it must be constantly replenished through diet or supplementation. Pantothenic acid is synthesized by plants, fungi, and bacteria, so a well-balanced diet is a good source of pantothenic acid. Pantothenates have beneficial effects on the skin (regenerative, healing, and anti-inflammatory effects). They are readily absorbed through the skin, making them a common active ingredient in cosmetics and medications with soothing, healing, regenerative, moisturizing, and protective properties for damaged, sensitive, or atopic skin, including topical products labeled hypoallergenic or specifically intended for people with sensitive skin. Although PA is considered non-allergenic and safe, paradoxically, frequent exposure, especially to damaged skin, appears to lead to hypersensitivity to this substance. The pathogenetic mechanisms of pantothenate hypersensitivity have not been clearly defined. The main sensitization pathway is likely a delayed cellular mechanism (type IV, contact). However, other types of hypersensitivity, including immediate (type I) and mixed mechanisms, cannot be ruled out. PA allergy is considered rare and therefore difficult to diagnose. This is due to the unexpected sensitizing factor and the lack of standard diagnostic tests. Due to the likely contact nature of the allergy, patch testing (PT) with a cosmetic (drug) provided by the patient (“as is”) and PA (5% in petrolatum; 5% PET) appears to be the best approach. It is also suggested that PA (5% PET>) be included in the standard series of allergens (haptens) used in routine PT diagnostics. It appears that PA allergy is more common than currently believed, particularly in people with atopic skin and polysensitized individuals, who are the primary users of products containing pantothenate. It is possible that in these individuals, pantothenic acid may be more harmful than beneficial.

Pharmaceutics doi: 10.3390/pharmaceutics18070770

Authors: Jong-Min Kim Kyoung-Ah Kim Na-Young Yu Dae-Duk Kim Jeong Yeon Kang Seung-Ki Baek Jin-Woo Park Ji-Young Park

Background/Objectives: Semaglutide, a long-acting glucagon-like peptide-1 (GLP-1) analog for type 2 diabetes and obesity, requires sensitive and high-throughput bioanalytical methods to support pharmacokinetic studies. However, previously reported liquid chromatography–tandem mass spectrometry (LC–MS/MS) assays have been limited by lengthy run times (~18 min) and suboptimal sensitivity. This study aimed to develop and validate a rapid, sensitive LC–MS/MS method for quantifying semaglutide in plasma. Methods: Plasma samples (50 μL) were prepared by acetone-mediated protein precipitation followed by solid-phase extraction. Chromatographic separation was performed on a Cadenza CD-C18 MF column within 9 min, using positive electrospray ionization in multiple reaction monitoring mode with the transitions m/z 1029.4 → 110.1 for semaglutide and m/z 938.9 → 109.9 for liraglutide (internal standard). Validation followed the U.S. Food and Drug Administration (FDA) bioanalytical guidelines. Results: The assay showed a lower limit of quantification of 1 ng/mL with linearity across 1–500 ng/mL (R2 = 0.9999), with sharp peak shape and no carryover. Intra- and inter-day accuracies were 95.69–103.76% and 94.93–100.08%, with precision ≤4.50% and ≤5.88%. Recovery (93.05–107.95%) and matrix effects (96.34–104.12%) were consistent across quality control levels, and the analyte was stable under all tested conditions. The method was successfully applied to a pharmacokinetic study in Sprague–Dawley rats following subcutaneous administration of 50 μg semaglutide. Conclusions: The validated method offers shorter analysis time, improved sensitivity, and reduced sample volume compared with previously reported assays, supporting its application in preclinical pharmacokinetic studies of semaglutide and related GLP-1 analogs.

Pharmaceutics doi: 10.3390/pharmaceutics18070769

Authors: Kang Shu Yating Lei Linjie Li Shike Wang Ting Du Ting Tong

Gold nanoparticles (AuNPs) have emerged as versatile antiviral nanoplatforms because their size, morphology, plasmonic properties, and surface chemistry can be precisely engineered. In this review, we summarize the core design principles of antiviral AuNPs from a structure–function–mechanism perspective. We first outline representative synthetic and interface-programming routes for AuNP preparation, including citrate reduction, Brust–Schiffrin synthesis, seed-mediated growth, green synthesis, direct thiol-conjugation, and mixed-ligand shell strategies, emphasizing how these approaches define particle size, morphology, surface accessibility, interfacial composition, and downstream biofunctionalization potential. We then discuss major surface engineering strategies, including polyethylene glycol, nucleic acids, antibodies and nanobodies, peptides, glycans, antiviral drugs, and biomimetic coatings, with particular attention to how ligand density, orientation, flexibility, and interfacial stability determine biological performance. Next, we examine how functionalized AuNPs inhibit different stages of the viral life cycle, including viral attachment and entry, intracellular replication, assembly and egress, photothermal inactivation, and immune modulation or vaccine delivery. Finally, we highlight current challenges, including incomplete structure–activity relationships, dynamic nano–bio interactions under physiological conditions, limited standardization across studies, and translational barriers related to safety, reproducibility, and scale-up. This review provides a conceptual framework for the rational development of next-generation AuNP-based antiviral nanotherapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18070768

Authors: Aolin Chen Ping Luo Jing Cao Taohong Su Xinxin Ding Xinzhi Guo Wenhao Zhou Yang Chen Fang Wang

Traditional mineral drugs represent an underexploited reservoir of natural antitumor agents; however, their clinical translation has historically been hindered by poor bioavailability, non-specific biodistribution, and dose-limiting toxicity. This review comprehensively examines the pharmacological mechanisms and modern formulation strategies driving the renaissance of mineral-based oncology therapeutics. We highlight how mineral drugs exert potent anticancer effects through interconnected pathways, including regulated cell death (e.g., apoptosis, ferroptosis), cell-cycle arrest, and immunomodulation. Crucially, we evaluate recent advances in drug delivery systems, such as liposomes, polymeric nanoparticles, inorganic frameworks, and stimuli-responsive (e.g., pH, redox, enzyme) release systems that successfully overcome traditional pharmacological barriers. These bioengineering strategies not only improve solubility and tumor targeting but also significantly widen the therapeutic window, as evidenced by enhanced tumor suppression and reduced systemic toxicity in preclinical models. Despite this progress, challenges regarding in vivo chemical transformations and tumor heterogeneity remain. Ultimately, we propose a closed-loop “Composition–Mechanism–Delivery” design paradigm to guide future research, facilitating the translation of ethnopharmacological heritage into precision mineral-based therapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18070767

Authors: Sara Guibunda Tajú Amanda Beatriz da Silva Soares Patrícia Aoki Miyasato Rafaela Paula de Freitas Lenita de Freitas Tallarico Erika Mattos Stein Pio Colepicolo Eliana Nakano

Background/Objectives: Schistosomiasis, a parasitic disease caused by Schistosoma worms with freshwater snails as intermediate hosts, affects over 250 million people. The current control relies solely on praziquantel, which raises concerns on drug resistance and highlights the need for new therapeutic alternatives. Our bioprospection studies have focused on marine macroalgae as an unexplored source of antischistosomal metabolites with promising results. Guided by WHO recommendations to target both the parasite and its transmission vectors, this study aimed to investigate Ochtodes secundiramea to: (i) isolate active metabolites; (ii) evaluate the isolated compounds against adult worms and oviposition to identify leads for drug development; and (iii) perform an independent screening of their effects against the environmental transmission stages on cercariae and B. glabrata embryos. Methods: A dichloromethane extract of O. secundiramea was submitted to an NMR–biomonitored guided fractionation against Schistosoma mansoni adult worms. Active fractions were further purified through HPLC and characterized by 1H and 13C NMR spectroscopy to identify the isolated compounds. Results: Three halogenated monoterpenes were isolated: ochtodene 1 (4-bromo-1,6,8-trichloro-2,3-ochtodene), ochtodene 2 (2-chloro-1,6,8-tribromo-3,8-ochtodene), and the novel natural product ochtodene 3 [2,6-dibromo-4-(2-chloroethylidene)-1,1dimethylcyclohexane]. Ochtodene 1 was the primary active metabolite against Schistosoma mansoni adult worms, with IC50/96 h values of 47.2 and 46.1 µM for male and female worms respectively, and totally suppressed egg laying with 60 µM, while showing no toxicity toward human fibroblasts. Notably, all metabolites, including the novel ochtodene 3, caused 100% mortality in cercariae and embryos at low concentrations. Conclusions: The discovery of the novel ochtodene 3 and the identification of distinct leads for host treatment and transmission elimination position O. secundiramea as a promising source for integrated schistosomiasis control.

Pharmaceutics doi: 10.3390/pharmaceutics18070766

Authors: Sonya Salamone Rosalia Pellitteri Ilaria Ottonelli Elide Zingale Cinzia Cimino Barbara Ruozi Teresa Musumeci Rosario Pignatello

Background: Effective strategies for delivering neuroprotective agents to the brain remain a major challenge due to the poor solubility, rapid metabolism, and low bioavailability of promising molecules, such as 7,8-dihydroxyflavone (7,8-DHF). This small-molecule TrkB receptor agonist exhibits significant antioxidant, neuroprotective properties, and additional effects on metabolic regulation, but its therapeutic potential is limited by unfavorable pharmacokinetic characteristics. Nanotechnology-based delivery systems are increasingly explored to improve drug stability, enhance bioavailability, and facilitate direct nose-to-brain transport following intranasal administration. In this study, lipid nanoparticles encapsulating 7,8-DHF were developed using a fish-oil-based lipid core enriched with ω-3 polyunsaturated fatty acids (DHA and EPA) and naturally derived excipients, including soybean lecithin and ε-polylysine. Methods: The formulation was optimized using a Design of Experiments (DoE) approach based on a 23 full factorial design, evaluating drug concentration, lecithin concentration, and surfactant type (Pluronic® F127 or Tween® 80). The main formulation responses considered were particle size, polydispersity index (PDI), zeta potential, and encapsulation efficiency. Results: The optimized nanoparticles exhibited nanometric dimensions (<250 nm); spherical morphology, confirmed by TEM; low polydispersity (PDI < 0.3); and adequate encapsulation efficiency. Stability studies in simulated biological fluids indicated good physicochemical stability for up to 48 h, while interaction studies with mucin suggested a good interaction within the mucus environment. ROS scavenging capacity was confirmed through the DPPH chemical assay, and in vitro experiments on olfactory ensheathing cells, selected as a biologically relevant model for their anatomical localization along the olfactory pathway, showed reduced cytotoxicity of the encapsulated drug compared with the free form. Conclusions: Collectively, these results support the potential application of the developed nanoformulation in the intranasal delivery of 7,8-DHF.

Pharmaceutics doi: 10.3390/pharmaceutics18060765

Authors: Lizeth Geraldine Muñoz Yhors Ciro Andrés Felipe Chamorro

The increasing prevalence of antimicrobial resistance (AMR) has promoted the development of advanced biomaterials capable of overcoming the limitations of conventional antibiotics. In this context, metal nanoparticle hybrid hydrogels (MNHHs) have emerged as multifunctional platforms that integrate the high water-retention capacity and biocompatibility of hydrogels with the antimicrobial properties of metallic nanoparticles (MNPs). This review critically analyzes recent advances in the design, physicochemical properties, antimicrobial mechanisms, and biomedical applications of these systems. Current evidence demonstrates that MNHHs can achieve antimicrobial efficiencies above 98–99%, with minimum inhibitory concentrations as low as 0.78 µg mL−1 and inhibition zones of up to 25 mm against clinically relevant pathogens. Furthermore, the incorporation of MNPs significantly improves the mechanical properties of hydrogels and enables controlled and sustained metal ion release for periods of up to 14 days. Despite these promising results, important challenges remain regarding cytotoxicity, release control, the lack of experimental standardization, and the limited understanding of long-term biological effects. Overall, MNHHs represent a promising strategy for infection control, regenerative medicine, and controlled drug delivery; however, their clinical translation still requires the development of reproducible, safe, scalable, and highly biocompatible systems.

Pharmaceutics doi: 10.3390/pharmaceutics18060763

Authors: Junmin Li Longjie Li Fangbin Ding Meixia Chen Mengyue Hu Xiaoqiang Xiang Jing Tang

Background/Objectives: Cipepofol is a novel intravenous anesthetic whose pharmacokinetics (PK) may vary with dosing regimens, sampling sites, and physiological differences across populations. However, clinical PK data remain fragmented across study settings and are limited for special populations and individualized perioperative use, highlighting the need for a mechanistic modeling framework. This study aimed to develop and evaluate a physiologically based pharmacokinetic (PBPK) model for cipepofol across diverse populations. Methods: Clinical data from nine studies were included, comprising 371 subjects and 3521 plasma concentration measurements. The model was established in healthy adults using HSK3486-101, qualified using healthy-adult data from HSK3486-111 and anesthesia induction datasets, and extrapolated to hepatic impairment, renal impairment, and elderly populations using pathophysiology-informed adjustments. Individualized external validation was further performed in adult and pediatric surgical patients using actual clinical dosing histories. Model performance was evaluated using concentration–time profiles, goodness-of-fit plots, fold error, and geometric mean fold error (GMFE) for Cmax and AUC0–t. Results: The model adequately described both arterial and venous plasma concentration–time profiles across the establishment, qualification, extrapolation, and external validation datasets. Most predicted concentrations were within two-fold of the observed values, and the overall GMFE values were 1.22 for Cmax and 1.21 for AUC0–t. Simulated exposure differences in hepatic impairment, renal impairment, and elderly subjects were generally limited, suggesting no clinically meaningful PK changes from a PK exposure perspective in these populations. The model also reproduced arterial–venous concentration differences and supported the major contributions of UGT1A9 and CYP2B6 to cipepofol clearance. Conclusions: This PBPK model provides a mechanistic framework for characterizing cipepofol disposition and may inform future model-informed dosing studies.

Pharmaceutics doi: 10.3390/pharmaceutics18060764

Authors: Renato Sonchini Gonçalves

Artificial intelligence (AI) is increasingly influencing nanopharmaceutical development by supporting the transition from empirical formulation screening toward predictive, data-driven, and translationally oriented design. Nanocarrier-based therapeutics are governed by nonlinear relationships among material composition, physicochemical attributes, manufacturing parameters, biological identity, pharmacokinetics, toxicity, and therapeutic performance. In this review, we examine how AI can contribute to nanopharmaceutical development from predictive formulation design to clinical translation. We synthesize current applications of machine learning, deep learning, physics-informed modeling, hybrid mechanistic–AI approaches, and automated optimization workflows, with emphasis on critical quality attribute modeling, multi-objective optimization, design of experiments, quality-by-design, process analytical technology, digital twins, and continuous manufacturing. We also discuss applications involving nano–bio interactions, pharmacokinetics, toxicity, immunogenicity, and precision nanomedicine. AI-based approaches can support rational nanocarrier design, identify nonlinear formulation–property relationships, guide optimization, improve process understanding, and integrate heterogeneous experimental, biological, and manufacturing datasets across diverse nanopharmaceutical platforms. These methods are particularly relevant for modeling protein corona formation, cellular uptake, intracellular trafficking, biodistribution, pharmacokinetics, toxicity, immunogenicity, and patient-specific responses. However, translational implementation remains limited by fragmented datasets, inconsistent reporting standards, limited interpretability, insufficient external validation, uncertain predictions, poorly defined applicability domains, and evolving regulatory expectations for adaptive computational models. Overall, AI should be viewed not only as an optimization tool, but also as a translational framework connecting formulation science, biological prediction, manufacturing control, and clinical implementation. Future progress will depend on standardized data infrastructures, explainable and externally validated models, uncertainty quantification, applicability-domain definition, hybrid mechanistic–AI frameworks, regulatory-ready documentation, and clinically relevant case studies.

Pharmaceutics doi: 10.3390/pharmaceutics18060762

Authors: Zhuo Chen Huiwen Pang Youzhi Wu David M. Klyne Xuqiang Nie Pengfei Jiang Xinfei Wu Kong-Nan Zhao Felicity Y. Han

Background/Objectives: Chronic wounds remain a formidable clinical challenge due to the suboptimal efficacy of conventional delivery systems and therapeutics. Textilinin-1, a venom-derived Kunitz-type serine protease inhibitor, has previously established its profile as a potent hemostatic agent. However, its potential as a multifunctional biopharmaceutical for wound management remains largely untapped. This study evaluates the pharmacological effects of Textilinin-1 in preclinical models of cutaneous wound repair. Methods: We employed an integrated platform comprising bioinformatics, in vitro cellular assays, and in vivo murine excisional wounds and a pilot porcine proof-of-concept model to assess the wound healing-promoting effects of Textilinin-1 and explore associated cellular responses associated with key stages of the wound healing cascade. Results: Textilinin-1 was associated with multiple cellular responses relevant to tissue repair. It attenuated M1-like inflammatory activation and showed preliminary growth-inhibitory activity against Staphylococcus aureus under the tested conditions. Concurrently, it enhanced the proliferative and migratory capacity of fibroblasts, endothelial cells, and keratinocytes, which are key cellular targets for wound closure. In pre-clinical pilot porcine and rodent models, Textilinin-1 treatment was associated with accelerated wound contraction and improved structural tissue quality. Conclusions: Our findings provide preclinical evidence that Textilinin-1 may promote cutaneous wound repair and modulate cellular responses relevant to key stages of the wound healing cascade. These results support further investigation of Textilinin-1 as a candidate for wound repair applications. Future studies are required to define its precise molecular mechanisms, evaluate its efficacy in chronic or otherwise compromised wound models, and optimize its topical formulation or hydrogel-based delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18060761

Authors: Juan Liu Ziheng Wang Yuchang Yang Lisha Yi Shiman Li Jingyi Gao Jia Zhou Nannan Cheng Xingbin Yin Xiaoxv Dong Jian Ni Changhai Qu

Objectives: This study aimed to develop curcumin nanoparticles (Cur@PCL-PEG-MF/cRGDfc) with retinal-targeting capability and to evaluate their biological effects and pharmacological mechanisms in vitro. Methods: After synthesis of the carrier framework, metformin (MF) and cRGDfc were conjugated to the carrier material using the carbodiimide method and Michael addition reaction, respectively. Subsequently, self-assembled nanoparticles were formed from the carrier and curcumin under specific conditions. The materials were characterized by spectroscopy, chromatography, elemental analysis, energy-dispersive spectroscopy and X-ray diffraction. The efficacy of the formulation was evaluated in two cell lines, ARPE-19 and HUVEC-T1. In addition, the pharmacological mechanism was explored using transcriptome sequencing as a complementary approach. Key Findings: Self-assembled nanoparticles were successfully prepared by combining the two modified carrier materials, PCL-PEG-MF and PCL-PEG-cRGDfc, with curcumin. The nanoparticles exhibited an encapsulation efficiency of 78.09%, a particle size of 162.33 nm, and a zeta potential of −23.28 mV and displayed a spherical morphology. They showed sustained release in simulated physiological conditions and stronger affinity for ARPE-19 cells under oxidative stress. Nearly 100% of the nanoparticles were internalized by the cells, which was accompanied by reduced ROS and LDH release and decreased DNA fragmentation. In addition, the nanoparticles inhibited neovascularization by reducing VEGF-A release, thereby potentially protecting the retina in macular degeneration and reducing choroidal hemorrhage. Further analyses showed that curcumin and its nanoformulations significantly reduced the expression of inflammatory factors such as IL-1β and IL-18, lowered the protein levels of Caspase-1, GSDMD-N, and NLRP3, and increased AMPK levels. Conclusions: Using PCL-PEG as the carrier framework, MF and cRGDfc were conjugated to construct a curcumin-loaded nanoparticle with retinal-targeting capability. This nanoparticle, characterized by a small particle size, sustained release, and targeted delivery to retinal pigment epithelium (RPE) cells under oxidative stress, alleviated oxidative stress-induced damage. Its therapeutic effect may be mediated, at least in part, by interference with the AMPK/mTOR pathway and activation of the NLRP3/Caspase-1/GSDMD pathway.

Pharmaceutics doi: 10.3390/pharmaceutics18060760

Authors: Sizhen Wang Qiwei Tai Kehui Wang Jianyu Zheng Beibei Guo Feng Yang Chen Wang

Background/Objectives: Lipid nanoparticles (LNPs) have emerged as crucial vehicles for messenger RNA (mRNA) applications in antitumor therapy. Combining LNPs with stimulator of interferon genes (STING) activation holds promise for treating “cold” tumors such as pancreatic cancer. However, two major challenges remain: inefficient mRNA escape from endosomes and STING pathway suppression in immunosuppressive tumor microenvironments. Methods: To improve endosomal escape, we developed a novel pH-responsive PEGylated lipid (Ben-mPEG2000) for mRNA-LNP preparation while using commercial Man-mPEG2000 for dendritic cell (DC)-targeted delivery of LNPs; to alleviate suppression of the STING pathway in the tumor microenvironment and activate immune responses, STING-R283S mRNA was encapsulated into LNPs, ultimately resulting in DC-targeted/pH-responsive LNPs loaded with STING-R283S mRNA for pancreatic cancer immunotherapy research. Results: After pH-responsive cleavage, Ben-mPEG2000 not only enhanced the positive charge of LNPs through the exposed protonated amino groups but also eliminated the PEG-induced steric hindrance effect. The combination of these two effects promoted membrane fusion between LNPs and the endosome, thereby enhancing mRNA translation. As a payload, STING-R283S could further amplify STING signaling in DCs without cytotoxicity to counteract immunosuppression in pancreatic cancer. Conclusions: This engineered LNP platform enhanced mRNA expression and STING activation in DCs, improving immunotherapy outcomes in pancreatic cancer.

Pharmaceutics doi: 10.3390/pharmaceutics18060759

Authors: Aphelele Taliwe Siphamandla Q. N. Lamula Lisa V. Buwa-Komoreng Vuyolwethu Khwaza

The Asteraceae family represents one of the largest groups of medicinal plants, widely used in traditional medicine and increasingly investigated for its pharmacological potential. This review summarizes current evidence on the botanicals derived from Asteraceae plant species and their molecular mechanisms of action against inflammation and cancer. Major classes of bioactive compounds in extracts are discussed in relation to their modulation of key signaling pathways and therapeutic targets such as NF-κB, MAPK, PI3K/Akt, COX-2, iNOS, and apoptotic regulators (Bax/Bcl-2, caspases). A literature search covering studies published between 2022 and 2026 was conducted. Evidence from in vitro, in vivo, and in silico studies demonstrates that Asteraceae-derived botanicals exert therapeutic effects through antioxidant activity, cytokine suppression, enzyme inhibition, and regulation of gene expression. Overall, the mechanistic insights presented herein support the rational use of Asteraceae medicinal plants and identify promising lead compounds for drug discovery and development.

Pharmaceutics doi: 10.3390/pharmaceutics18060758

Authors: Hadi Shammout Béla Hopp Judit Kopniczky Tamás Smausz Bence Sipos Katalin Kristó János Bohus Orsolya Jójárt-Laczkovich Flórián Benkő Tamás Sovány Krisztina Ludasi

Background/Objectives: With the increasing incidence of drug counterfeiting and the emergence of personalized medicine, the need for unique marking of solid dosage forms, e.g., tablets, has attracted considerable interest in the current research and development landscape. Besides traditional printing methods, laser marking offers several advantages, as it eliminates the need for organic solvents and enables the generation of precise patterns. However, laser exposure may raise safety concerns regarding the stability of photosensitive drugs in the irradiated dosage forms. Therefore, the aim of the present study was to test the photoprotective effect of titanium dioxide (TiO2) and its various alternatives, e.g., talc, calcium carbonate (CaCO3), zinc oxide (ZnO), and black iron oxide (Fe3O4), alongside a ready-to-use reference formulation, Opadry® Brown, which contains TiO2 (titanium-containing, TC) on nifedipine, a light-sensitive model drug. Methods: Laser marking or short-term laser ablation at different wavelengths (193 nm, 248 nm, 532 nm, and 781 nm) was applied to different coating formulations. As a positive control, prolonged exposure to daylight was applied. The properties and photostability of these formulations were evaluated using several analytical methods (i.e., surface profilometry, Raman spectroscopy, and high-performance liquid chromatography (HPLC)). Results: The TiO2, ZnO, Fe3O4, and Opadry® TC Brown coatings maintained their color during the long-term study under all conditions. Furthermore, the prepared formulations exhibited different ablation depths and morphological changes depending on the coating and laser type. HPLC measurements confirmed significant differences in the protective ability of various pigments against sunlight and different types of lasers. Nevertheless, the obtained Raman spectra were not in complete agreement with HPLC results, which can be attributed to spectral overlap between key nifedipine degradation markers and excipient signals in the tablet core. Conclusions: Overall, laser treatment of tablets containing photosensitive drugs may induce API decomposition; however, this effect can be minimized or avoided by careful selection of the appropriate combination of laser type and photoprotective pigment. Under the applied experimental conditions, Ti:Sa laser treatment was associated with the lowest degree of nifedipine degradation among all formulations, while ZnO-containing coatings demonstrated the most consistent photoprotective performance against the majority of the tested laser types, while Fe3O4-containing coatings provided superior protection during prolonged sunlight exposure and Nd:YAG laser irradiation.

Pharmaceutics doi: 10.3390/pharmaceutics18060757

Authors: Sosuke Yoneda Daisuke Uta Kana Yasufuku Takuya Yamane Saho Yoshioka Keiko Takasu Takaya Izumi Sayaka Fujita Daiki Nakamori Shiori Kawasaki Tatsuya Takahashi Mai Yoshikawa Koichi Ogawa Erika Kasai

Background: Neuropathic pain remains difficult to treat because current analgesics often provide insufficient efficacy or dose-limiting adverse effects. Nav1.7 is genetically validated as a key regulator of human pain sensation, but the development of selective small-molecule Nav1.7 inhibitors has been limited by the high similarity among voltage-gated sodium channel subtypes. Methods: We generated monoclonal antibodies selectively targeting Nav1.7, humanized them for therapeutic development, and evaluated their binding, selectivity, functional channel inhibition, systemic analgesic efficacy, and effects on neuronal activity in a rat model of partial sciatic nerve ligation-induced neuropathic pain. Results: The humanized antibodies showed high-affinity and selective binding to Nav1.7 and functionally inhibited the channel in cellular assays. After systemic administration to neuropathic pain model rats, the lead antibody produced robust analgesia lasting at least 96 h. Electrophysiological analyses demonstrated reduced mechanically evoked and spontaneous neuronal activity, and immunohistochemistry showed decreased mechanical stimulus-induced phosphorylation of extracellular signal-regulated kinase in dorsal root ganglion neurons. The antibodies did not impair physiological nociception or motor function under the tested conditions. Conclusions: These findings provide preclinical proof of concept that humanized anti-Nav1.7 antibodies can act as systemically administered, long-acting biologic analgesics for neuropathic pain while preserving normal nociceptive and motor functions. The clinical advancement of S-151128 further supports the translational potential of this modality.

Pharmaceutics doi: 10.3390/pharmaceutics18060755

Authors: Faisal Khaled Aldawood

Additive manufacturing has emerged as a transformative technology for fabricating complex drug-eluting medical devices, offering unprecedented design freedom and functional integration capabilities. This comprehensive review systematically analyzes 3D printing technologies applied to pharmaceutical device manufacturing, focusing on drug-eluting vascular stents as a representative application. This review covers six primary additive manufacturing techniques, ranging from high-resolution vat photopolymerization (25 μm resolution) to direct energy deposition, with a focus on their capabilities for produce pharmaceutical devices with controlled drug release properties. Novel 4D/5D/6D printing technologies introduce stimuli-responsive behaviors enabling programmable drug release profiles and adaptive device functionality. Manufacturing process optimization reveals superior design flexibility compared to conventional methods, with 85–95% reduction in design iteration time and elimination of tooling costs for complex geometries. The material landscape encompasses traditional metals (316L stainless steel, cobalt–chromium), biodegradable polymers (polylactic acid, PLA; polycaprolactone, PCL; poly(lactic-co-glycolic acid), PLGA), shape-memory materials (i.e., polymers and alloys capable of recovering a pre-programmed shape upon exposure to a specific stimulus such as body temperature, moisture, or light), and advanced nanocomposites, each offering distinct drug-loading capacities (100–500 μg/cm2) and release kinetics. Critical challenges include standardization requirements (International Organization for Standardization (ISO) 5840 and American Society for Testing and Materials (ASTM) F2606), pharmaceutical-grade manufacturing protocols, and regulatory pathways for novel drug-device combinations. This review identifies key research priorities including development of biocompatible printing materials, accelerated drug release testing protocols, and scalable manufacturing processes suitable for medical device production. This analysis demonstrates that 3D printing enables integration of multiple pharmaceutical functions within single devices, controlled spatiotemporal drug delivery, and elimination of secondary manufacturing steps for drug coating processes, advancing the development of next-generation therapeutic medical devices.

Pharmaceutics doi: 10.3390/pharmaceutics18060756

Authors: Hao Ren Mengchen Han Yuchao Qiao Yu Cui Chongqi Hao Yiming Lou Gaomin Jing Qiankun Liu Lang Yang Li Zheng Lixia Qiu

Objectives: An integrated methodological framework was developed for modeling and multiobjective optimization of sustained-release drug delivery systems. Methods: The cumulative release percentage was fitted as a function curve, and functional principal component analysis was subsequently used to transform the function curves into functional principal component scores (FPCs). FPCs were then treated as dependent variables, while the proportions of the formulation factors were used as independent variables to construct Scheffé polynomial regression models. Finally, Non-dominated Sorting Genetic Algorithm III (NSGA-III) was applied to perform multi-objective optimization. Results: FPC1, FPC2, and FPC3 captured 95.18%, 4.39%, and 0.32% of the total variation, respectively. Corresponding Scheffé polynomial regression models were established, including quadratic models for FPC1 (adjusted R2 = 0.751, AIC = 168.557) and FPC2 (adjusted R2 = 0.592, AIC = 119.302), and a special cubic model for FPC3 (adjusted R2 = 0.597, AIC = 64.574). The NSGA-III algorithm generated a Pareto optimal set, yielding stable formulation compositions with mean (SD) values of X1 = 0.123 (0.015), X2 = 0.821 (0.032), X3 = 0.012 (0.017), and X4 = 0.045 (0.015). The corresponding FPCs were −41.787 (2.544), 10.009 (0.168), and 8.264 (0.010) for FPCs1–FPCs3, respectively. The reconstructed cumulative release percentages were 42.471 (1.661), 52.623 (2.868), 69.942 (1.200), 84.275 (1.010), and 93.330 (0.832), demonstrating good agreement with the target release profiles. Conclusions: The integrated FDA–Scheffé–NSGA-III framework provides a robust and effective approach for accurately modeling release behavior and optimizing sustained-release formulations.

Pharmaceutics doi: 10.3390/pharmaceutics18060754

Authors: Zlatina Vlahova Lazar Lazarov Maria Petrova Shazie Yusein-Myashkova Jordana Todorova Maria Schröder Monika Mutovska Stanimir Stoyanov Yulian Zagranyarski Iva Ugrinova

Background/Objectives: DNA-targeting small molecules that induce replication stress represent a promising strategy in anticancer drug development. 1,8-Naphthalimide (NI) derivatives are well-established DNA-intercalating agents, and heterocyclic annulation offers a rational approach to enhancing their potency and tumor selectivity. Here, we report the synthesis and biological evaluation of a novel series of benzofuran-containing naphthalimide derivatives, with particular focus on the lead dinitro-substituted compound 5d. Methods: Cytotoxic activity was assessed using the MTT assay in A549 (p53 wild-type), H1299 (p53-null), and MRC-5 cells. Long-term antiproliferative effects were evaluated by clonogenic survival assay. Cell cycle distribution was analyzed by propidium iodide staining and flow cytometry. Replication stress and DNA damage were quantified by EdU incorporation and γH2AX immunofluorescence, respectively. Apoptosis was assessed by Annexin V/PI staining and caspase-3/7 activation assay. p53 nuclear accumulation and autophagy induction were evaluated by immunofluorescence and Western blot, using LC3 as an autophagic marker. Results: All compounds exhibited cytotoxic activity in the nanomolar range, with 5d emerging as the most potent and selective. Clonogenic survival was significantly reduced, indicating durable suppression of proliferative capacity. Treatment with 5d induced G1 arrest in A549 cells and the accumulation of H1299 cells in G2/M, consistent with p53-dependent and p53-independent checkpoint activation, respectively. EdU incorporation was markedly reduced, while γH2AX intensity increased, collectively supporting a replication stress-driven mechanism of DNA damage. Apoptosis was confirmed by increased Annexin V-positive populations and caspase-3/7 activation. LC3 puncta formation and LC3-I/LC3-II conversion were increased, indicating LC3 processing and autophagosome accumulation consistent with the activation of autophagy-related processes. Conclusions: 5d induces a cellular phenotype consistent with replication stress, including reduced EdU incorporation, γH2AX accumulation, cell cycle arrest, and apoptotic cell death in a p53 status-dependent manner. These findings establish benzofuran-annulated naphthalimides as a promising scaffold for the development of anticancer agents that exploit replication stress vulnerabilities in tumor cells.

Pharmaceutics doi: 10.3390/pharmaceutics18060753

Authors: Mashan Almutairi Ahmed Adel Ali Youssef Yazed S. Alsowaida Ahmed Alobaida Samir A. Ross

Background/Objective: Topical therapy remains a cornerstone in managing fungal infections due to the deep-seated nature of the pathogens and the persistence of the disease. Ketoconazole (KTZ) is a broad-spectrum antifungal agent, but its highly lipophilic nature presents considerable challenges in developing effective topical formulations. Additionally, oral KTZ has been subject to labeling restrictions and market withdrawal due to its association with severe hepatic adverse effects. This study was conducted to design, optimize, and evaluate KTZ-loaded nanolipid carriers (NLCs; KTZ-NLCs) as a delivery platform that could improve cutaneous bioavailability and enhance antifungal activity. Methods: The optimized KTZ-NLCs were further incorporated into a mucoadhesive system (KTZ-NLCs-C) through the inclusion of Carbopol® 940 NF, aiming to improve the retention of the formulation on the skin surface. NLCs were characterized in terms of their physical appearance, particle size, polydispersity index, zeta potential, pH, viscosity, drug content, and entrapment efficiency. The optimized KTZ-NLC and KTZ-NLCs-C formulations were subsequently assessed for in vitro drug release, ex vivo skin permeation and deposition, as well as in vivo skin irritation. Results: In vitro release studies revealed that nanocarrier systems provided a sustained release of KTZ over 24 h. The ex vivo transdermal flux and permeability coefficient of KTZ from the lead KTZ-NLCs-C formulation were approximately 2.8-fold greater than those achieved with the marketed cream formulation. The in vivo skin irritation studies indicate that NLC-based formulations are suitable for topical applications. The lead formulation was stable for 90 days (the final time point evaluated) under refrigerated and room-temperature storage conditions. Conclusions: These findings suggest that the NLC-based system is a promising platform for the topical delivery of KTZ and has the potential to enhance the therapeutic outcomes for patients with superficial fungal infections.

Pharmaceutics doi: 10.3390/pharmaceutics18060752

Authors: Dan Li Jiao Lu Qianru Li Huan Deng Songwei Tan

Background/Objectives: Immune checkpoints are critical regulatory pathways that maintain peripheral tolerance and prevent autoimmunity. Among these, the programmed death-1/programmed death-ligand 1 (PD-1/PD-L1) axis serves as a major inhibitory pathway that terminates T cell responses. While protein-based checkpoint blockade (ICB) targeting this axis has revolutionized clinical cancer therapy, its clinical efficacy is frequently limited by low response rates, immune-related adverse events (irAEs), and the emergence of adaptive resistance. To break through these bottlenecks, genetic interruption has emerged as a high-precision alternative to modulate the PD-1/PD-L1 pathway at the nucleotide level. Methods: A comprehensive systematic review of literature was performed across major databases (PubMed, Web of Science), with a focus on high quality studies published up to 2026. Results: Direct genomic disruption via CRISPR/Cas9 and post-transcriptional silencing through RNA interference can effectively neutralize inhibitory signaling at its source. Recent advances demonstrate that targeting upstream regulatory nodes—including metabolic checkpoints (e.g., lactate metabolism) and biophysical mechanisms (e.g., liquid–liquid phase separation)—provides superior transcriptional control over PD-L1. Furthermore, engineering CAR-T cells with multiplex gene editing (e.g., TCR/B2M/PD-1 knockout) or localized scFv secretion significantly enhances antitumor potency while reducing systemic toxicity. Innovations in organ-targeted lipid nanoparticles and stimuli-responsive biomimetic carriers further address the delivery barriers in solid tumors. Conclusions: Gene therapy provides a high-precision platform for PD-1/PD-L1 modulation, offering a viable strategy to overcome adaptive resistance. Future clinical application depends on the refinement of safer editing tools, such as base editing, and the standardization of intelligent delivery systems to ensure controllable and scalable cancer immunotherapy.

Pharmaceutics doi: 10.3390/pharmaceutics18060751

Authors: Bac V. G. Nguyen Hoai Thu Le Tien-Trung Dao Quy-Nguyen Doan Duc-Huy Pham Nghi Bao Nguyen Minh-Tri Le Du-Thien Nguyen Phuoc-Vinh Nguyen

Background/Objectives: Otomycosis is a recurrent fungal infection of the external auditory canal. This disease is difficult to manage with current antifungal agents due to irritation, ototoxicity risk, and emerging resistance. Natural essential oils have been proposed as alternatives, yet their clinical application in otic formulations remains limited due to their poor solubility and stability. In this study, we report the first ear-drop formulation combining microemulsified Ocimum gratissimum essential oil and acetic acid for otomycosis treatment. Methods: The essential oil was quality-validated with eugenol content superior to 60%. A systematic formulation study was performed, and the Tween 20/isopropanol (4:1, w/w) mixture was selected as the optimal surfactant system, yielding a stable microemulsion with high encapsulation efficiency (~98%) and relevant physicochemical stability (up to 28 days). The final formulation containing 1% acetic acid and 0.3% micro-emulsified essential oil met pharmacopeial requirements in terms of appearance, pH, viscosity, and microbial limits. Results: Importantly, this micro-emulsified eardrop demonstrated significantly greater in vitro antifungal activity than 3% boric acid and 2% acetic acid eardrops in twelve clinical fungal isolates from Vietnamese swimmers, especially on Curvularia, Cunninghamella, Aspergillus terreus, and Bipolaris. Although less pronounced than 1% clotrimazole, the finalized formulation demonstrates better antifungal kinetics and a broader activity spectrum. Conclusions: This work provides relevant experimental evidence on the use of Ocimum gratissimum essential oil in a microemulsion delivery system and demonstrates its efficacy against clinically relevant otomycosis pathogens. The results establish a foundation for future in vivo and clinical studies.

Pharmaceutics doi: 10.3390/pharmaceutics18060750

Authors: Huan Deng Yi-Wen Zhang Qian-Fu Zhao Zhi-Jun Huang

Plant-derived exosome-like nanoparticles (PELNs), a subset of extracellular vesicle (EV) secreted by plant cells, have emerged as revolutionary biomaterial with broad applications in biomedicine, agriculture, and nanotechnology. Structurally, PELNs feature a phospholipid bilayer homologous to plant cell membranes, encapsulating bioactive components such as proteins, nucleic acids, lipids, and secondary metabolites. The native structure of PELNs endows them with enhanced bioavailability, reduced immunogenicity, and improved barrier penetration for precise tissue delivery. Recent studies highlight the cross-kingdom therapeutic potential of PELNs in mammals, including antitumor, anti-inflammatory, tissue repair, immunomodulation and so on. This review comprehensively summarized recent advancements in PELN research, including innovative isolation techniques, molecular characterization, their roles in drug delivery and disease therapy. We also discussed challenges in standardization, scalability, and regulatory frameworks which could provide future perspectives for translating PELNs into clinical and industrial applications.

Pharmaceutics doi: 10.3390/pharmaceutics18060749

Authors: Tomasz Todryk Monika Budnicka Lukasz Pajchel Hanna Kierońska Maciej Dawidowski Krzysztof Adam Stępień Joanna Giebułtowicz Sebastian Granica Joanna Kolmas Jakub P. Piwowarski

Background: 5-(3′,4′-Dihydroxyphenyl)-γ-valerolactone (DHPV) is a postbiotic gut microbiota-derived flavanol metabolite with reported anti-inflammatory activity. Despite growing interest in its potential dermatological applications, its pharmaceutical properties and suitability for topical delivery have not been systematically investigated. This study aimed to perform the first comprehensive preformulation and formulation-oriented evaluation of DHPV and to develop stable topical ointment formulations suitable for further dermatological research. Methods: The physicochemical properties of DHPV were characterized using powder X-ray diffraction (PXRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), quantitative solubility assessment, and excipient compatibility studies. Based on the obtained preformulation data, two anhydrous ointment formulations containing DHPV were developed. The formulations were evaluated for homogeneity, rheological behavior, chemical stability under accelerated storage conditions, and in vitro drug release performance. Results: DHPV was identified as a crystalline compound with heterogeneous particle morphology and limited aqueous solubility. Quantitative solubility studies demonstrated the highest solubility in PEG 300 and glycol-based solvents. Compatibility testing revealed increased impurity formation in hydrophilic environments, whereas lipophilic excipients provided improved chemical stability. Both ointment formulations exhibited acceptable physical characteristics and maintained DHPV stability throughout accelerated storage. However, marked differences in release behavior were observed. The lipid–wax formulation showed significantly higher release rates, lower variability, and more reproducible release profiles than the petrolatum-based reference formulation, indicating more efficient diffusion of DHPV from the semisolid matrix. Conclusions: The physicochemical characteristics of DHPV strongly influence formulation design and performance. Anhydrous lipid-based systems provide a favorable environment for maintaining DHPV stability, while formulation composition significantly affects drug release. The developed lipid–wax formulation represents a promising platform for future skin permeation, pharmacodynamic, and efficacy studies.

Pharmaceutics doi: 10.3390/pharmaceutics18060748

Authors: Adekunle Oduneye Odularu Anuoluwapo Temitope Adesegun Chukwuemeka Paul Azubuike Oluwadamilola Miriam Kolawole

Background/Objectives: Oral candidiasis is an infection of the oral cavity caused by Candida albicans. Mucoadhesive buccal films could adhere to the buccal mucosa for prolonged periods, improving the therapeutic outcomes of patients with oral candidiasis. This study aimed to develop and evaluate the properties of fluconazole containing sodium alginate/methylcellulose-based buccal films for potential treatment of oral candidiasis. Methods: Drug-polymer compatibility was investigated using FT-IR spectrophotometry. Three optimised fluconazole films (F1 to F3) containing 1–1.6% sodium alginate and methylcellulose (1.6%) were formulated using the solvent-casting method. Their physicomechanical properties were characterised using standard protocols. Drug content and in vitro drug release profiles were evaluated using UV-visible spectroscopy; in vitro/ex vivo mucoadhesion studies were conducted using the shaking water bath technique, and their antifungal activity against Candida albicans was evaluated using the agar ditch method. Results: FT-IR data analysis revealed that sodium alginate, methylcellulose and fluconazole were compatible in the films. The films were off-white, smooth, peelable, thin, with satisfactory pH values, folding endurance, drug content, excellent zones of inhibition against Candida albicans (40 mm), controlled drug release profile (3.6–4.1 mg/cm2 after 6 h), and they displayed Korsmeyer–Peppas drug release kinetics. Film F3 containing 1.6% sodium alginate and 1.6% of methylcellulose exhibited superior swelling index (70 ± 1%), tensile strength (0.68 ± 0.04 MPa) and in vitro/ex vivo mucoadhesion time (5.5 ± 0.3 h; 2.3 ± 0.3 h) relative to other studied films. Conclusions: The sodium alginate content of the films influenced their tensile and mucoadhesive properties. Film F3 was the most promising formulation for potential treatment of oral candidiasis.

Pharmaceutics doi: 10.3390/pharmaceutics18060747

Authors: Gabriela Adriany Lisboa Zilli Samara Cristina Mazon Patricia Viera de Oliveira Felipe Zaniol Eulália Lopes da Silva Barros Ângela Maria Lodi Chaiane Lunelli Saretto Hemyly Cardoso Ana Lúcia Anversa Segatto Sara Marchesan Oliveira J. Vladimir Oliveira Indiara Brusco

Background/Objectives: Paclitaxel is a chemotherapy drug used to treat various tumours, but its use is often limited by an acute and chronic pain syndrome that is poorly managed. Naringin and its aglycone, naringenin, exhibit antioxidant, antitumour, anti-inflammatory, and antinociceptive effects, making them potential alternative treatments. However, their low water solubility limits their oral bioavailability in humans. Micronisation in a supercritical medium reduces particle size and enhances the dissolution of compounds, offering a possible solution. In this study, we investigated whether micronising naringin and naringenin via supercritical technology could improve their dissolution and oral efficacy against paclitaxel-induced pain syndrome. Methods: Micronisation was performed using supercritical CO2. Molecular docking was used to analyse the binding of naringin and naringenin to TRPV1, a key target for pain relief. Swiss mice were used in capsaicin (TRPV1 agonist)-induced nociception and paclitaxel-caused acute and chronic pain models. We assessed mechanical, cold, and heat sensitivity, potential adverse effects, and TRPV1 mRNA expression. Results: Micronisation improved the apparent dissolution profile of molecules. Docking results showed that naringin and naringenin bind to TRPV1. Both micronised compounds reduced capsaicin-induced nociception without affecting locomotion or body temperature. Micronised naringin and naringenin alleviated mechanical and cold allodynia, as well as thermal hyperalgesia in both acute and chronic paclitaxel-induced pain, outperforming their conventional forms. They also downregulated TRPV1 mRNA expression in the mice’s sciatic nerve. Conclusions: Taken together, these results show that supercritical micronisation improved the apparent dissolution and oral antinociceptive efficacy of naringin and naringenin, emphasising their potential as promising alternatives for managing paclitaxel-induced pain, with TRPV1 being a probable contributor to the observed antinociceptive effects.

Pharmaceutics doi: 10.3390/pharmaceutics18060746

Authors: Magdalena Paczkowska-Walendowska Elwira Sieniawska Zbigniew Krasiński Judyta Cielecka-Piontek Krystyna Skalicka-Woźniak

Background: Thymoquinone (TQ), a bioactive compound derived from Nigella sativa L., exhibits promising antioxidant, anti-inflammatory, and wound-healing properties; however, its clinical application is limited by poor solubility and instability. Methods: In this study, three electrospun nanofiber systems based on different polymeric matrices, PVP (N1), PVP/HPβCD (N2), and PVP/PCL (N3), were developed as potential wound dressing materials for controlled TQ delivery. Results: All formulations produced uniform nanofibrous structures with TQ molecularly dispersed within the polymer matrix, as confirmed by SEM, XRPD, and FTIR analyses. The composition of the nanofibers significantly influenced their physicochemical and functional properties. The N2 system, containing hydroxypropyl-β-cyclodextrin (HPβCD), exhibited the smallest fiber diameter (~208 nm), the fastest drug release, and enhanced antioxidant and anti-inflammatory activity due to improved TQ solubility. In contrast, the N3 system, incorporating polycaprolactone (PCL), formed thicker fibers (~1089 nm) and demonstrated sustained release behavior, the highest mucoadhesion, and the most pronounced wound-healing effect (90% closure after 24 h). Stability studies revealed that HPβCD significantly improved TQ resistance to thermal, humidity, and photolytic degradation, whereas the PVP-based system without stabilizers showed the lowest stability. Principal component analysis (PCA) confirmed that nanofiber performance is governed by two key factors: drug availability and sustained release combined with bioadhesion. Importantly, wound-healing efficiency correlated more strongly with the latter. Conclusions: The results demonstrate that rational design of polymer composition enables modulation of TQ delivery and biological response. Among the tested systems, PVP/PCL nanofibers appear to be the most promising candidates for wound-dressing applications due to their ability to provide sustained drug release and enhance tissue regeneration.

Pharmaceutics doi: 10.3390/pharmaceutics18060745

Authors: Anna Shao Jingyan Zhang Zhaowei Jin Yao Li Jialin Tang Quanmin Chen Hongbing Wu Jeremy Guo

Nanoemulsion (NEM) is an effective adjuvant and delivery system for vaccines and nucleic acids, capable of inducing immune responses against diverse pathogens. Background/Objectives: Conventional NEM manufacture uses multi-step operations, typically high-shear homogenization and then microfluidization (HSHM), thereby increasing process complexity and contamination risk. As water-rich colloidal dispersions, NEM is prone to microbial proliferation and droplet coalescence; freezing further disrupts microstructure, causing phase fusion and separation, so NEM adjuvants are often stored separately from antigens in multi-vial formats. Lyophilization could reduce cold-chain dependence and enable single-vial products, but there is no systematic study on lyoprotectants comparation and process optimization of lyophilized NEM. Methods: An impingement jet mixing (IJM) process was evaluated as a simplified, scalable route for NEM production. Key IJM parameters, including flow ratio, total flow rate, preparation temperature, microchannel type, and shear mode—were examined to match attributes of conventional HSHM. Lyophilized and reconstituted NEM were characterized by dynamic light scattering, scanning electron microscopy, transmission electron microscopy, differential scanning calorimetry and/or in vitro potency to inform lyoprotectant selection, and Taguchi Design of Experiment (DOE) methodology guided lyophilization processes. Results: IJM yielded NEM with droplet size, polydispersity index (PDI) and morphology comparable to HSHM, with higher throughput and fewer unit operations. Optimized lyophilization technique with designed lyoprotectant and process formed closed structures to prevent the easy-to-flow monolayer of the emulsion from fusing, producing robust and stable NEM. Conclusions: Coupling IJM with targeted lyophilization establishes a scalable, lower-risk manufacturing paradigm for NEM that preserves critical quality attributes, reduces cold-chain reliance and enables single-vial adjuvanted vaccine formats with tangible industrial and clinical benefits.

Pharmaceutics doi: 10.3390/pharmaceutics18060744

Authors: Eungyeop Lee Jum Bum Kwon Hyuk Jun Cho Mi Ran Woo Dong Wuk Kim Jong Oh Kim Duhyeong Hwang

Oral administration remains the most convenient and favored route for systemic delivery of small-molecule drugs, primarily due to patient compliance and the absence of invasive procedures. Yet, poor aqueous solubility, chemical/enzymatic instability, and limited permeability in the gastrointestinal (GI) tract often result in low bioavailability (BA) of many therapeutic agents. Polymeric micelles formed from the self-assembly of amphiphilic block copolymers have gained considerable attention as a nanotechnology-driven solution to overcome these challenges. Their hydrophobic core–hydrophilic shell structure enables efficient encapsulation of poorly soluble small molecule drugs, providing protection from acidic or enzymatic degradation while potentially enhancing drug transport across the intestinal epithelium. This review examines the design principles, formulation strategies, and in vivo performance of polymeric micelles for oral delivery of small molecule drugs. We discuss strategies to improve micelle stability in the GI environment, including optimization of core hydrophobicity, kinetic stabilization, and corona engineering, and compare polymeric micelles with established alternatives such as self-micro emulsifying drug delivery system (SMEDDS) and amorphous solid dispersions (ASDs) across critical performance parameters. Despite decades of preclinical progress, no oral polymeric micelle formulation has reached regulatory approval, underscoring the persistent challenge of maintaining micellar structural integrity under the dynamic conditions of the GI environment. This review therefore examines not only the promise but also the structural vulnerabilities of oral micelles, proposing a stability-centered framework for interpreting micelle function under GI conditions. Finally, we discuss current translational challenges and suggest directions for future research toward clinical application of oral polymeric micelle systems.

Pharmaceutics doi: 10.3390/pharmaceutics18060743

Authors: Huan Zhang Yangyang Wang Jihan Wang Hui Li

Small-molecule drug development faces high attrition rates driven by complex pharmacokinetics and unforeseen toxicities. While deep learning offers high predictive accuracy, its opaque “black-box” nature hinders mechanistic transparency, clinical trust, and regulatory approval. This review synthesizes how Interpretable Machine Learning, synergized with systems pharmacology, advances this paradigm by enhancing mechanistic transparency in drug development. By providing insights into algorithmic decisions, Interpretable Machine Learning helps researchers identify molecular features that are statistically associated with absorption, distribution, metabolism, and excretion optimization and preemptively mitigate toxicophores, while noting that these associations require experimental validation to establish genuine causality. Furthermore, integrating multi-omics data via Interpretable Machine Learning guides rational polypharmacology, bridging in silico target identification with “dry-wet loop” validations. Crucially, Interpretable Machine Learning accelerates clinical translation by discovering causal biomarkers, refining patient stratification, and generating transparent “Model Cards” to satisfy U.S. Food and Drug Administration/European Medicines Agency regulations. We also discuss future challenges: data heterogeneity, out-of-distribution generalizability, and the evolution toward Causal Artificial Intelligence. Ultimately, the integration of Interpretable Machine Learning provides a framework for more transparent and evidence-based drug design, realizing the promise of precision medicine.

Pharmaceutics doi: 10.3390/pharmaceutics18060742

Authors: Sadettin Berkay Sarli Asiye Busra Boz Er

Magnetic fields (MFs) represent an emerging modality in cancer therapy, encompassing static, low-frequency, pulsed, and nanoparticle-mediated alternating fields. These interventions have demonstrated the capacity to modulate proliferation, apoptosis, ferroptosis, migration, and epithelial-to-mesenchymal transition (EMT) in tumor cells, often through reactive oxygen species (ROS) modulation, ion channel regulation, membrane receptor dynamics, and lysosomal membrane permeabilization. Magnetic nanoparticle hyperthermia (MHT) has reached clinical application, showing promising outcomes in glioblastoma and prostate cancer, while pulsed electromagnetic fields (PEMFs) and magneto-mechanical approaches are under preclinical investigation. The mechanistic diversity of MFs allows synergistic combination with chemotherapy, radiotherapy, and immunotherapy. However, parameter sensitivity, field standardization, and long-term safety remain challenges. Here, we review mechanistic insights, signaling pathways, and experimental and clinical evidence for MF-based cancer therapies, highlighting translational potential and the need for rigorous optimization to realize clinical efficacy.

Pharmaceutics doi: 10.3390/pharmaceutics18060741

Authors: Justyna Baranowska Anna Kiss Łukasz Szeleszczuk

Dipeptidyl peptidase-4 (DPP-4) is an established therapeutic target in the treatment of type 2 diabetes mellitus (T2DM), primarily due to its role in regulating incretin activity and glucose homeostasis. Although clinically approved DPP-4 inhibitors are widely used, their moderate efficacy has driven the search for novel compounds with improved properties. In this context, natural products have attracted considerable attention as a source of structurally diverse and biologically active molecules. At the same time, molecular docking has emerged as a key computational tool for the identification and evaluation of potential DPP-4 inhibitors. This review summarizes and critically analyzes current molecular docking studies of natural compounds targeting DPP-4. Over 150 studies were evaluated with respect to docking methodologies, selection of protein structures, and validation strategies. The results reveal substantial variability in computational protocols. Frequently used protein structures include ligand-bound DPP-4 models such as 1X70 and 6B1E. Among the investigated compounds, flavonoids represent the most extensively studied class, followed by alkaloids, phenolics, terpenoids, and peptides. Despite numerous reports of favorable binding interactions within the DPP-4 active site, many studies rely solely on docking results without further validation. The limited use of molecular dynamics simulations and experimental assays highlights a significant gap in the current literature. Overall, while molecular docking provides valuable preliminary insights, improved standardization and integration with complementary approaches are essential to enhance the reliability and translational relevance of in silico findings.

Pharmaceutics doi: 10.3390/pharmaceutics18060740

Authors: Otman Saud Dallal Blidi Emily Hayes Celine Souilhol Rawan Maani Alice Johnson Keith Chapple Nick Peake

Background/Objectives: Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, with poor outcomes in advanced stages and significant limitations in current chemotherapy regimens due to systemic toxicity. Extracellular vesicles (EVs) have emerged as promising natural drug delivery vehicles, offering the potential for targeted, less toxic therapies. This study investigates the feasibility of using autologous, patient-derived EVs as a delivery system for the chemotherapeutic agent doxorubicin, focusing on how disease stage and the EV protein corona influence loading and delivery efficiency. Methods: EVs were isolated from plasma and tissue samples of CRC patients at different disease stages, as well as from healthy controls, demonstrating successful isolation and characterisation of EVs, with distinct profiles across different sources. Results: Doxorubicin loading into EVs was significantly higher in CRC patient-derived EVs compared to healthy controls, and tissue-derived EVs yielded higher quantities of drug-loaded particles. Delivery of doxorubicin-loaded EVs to recipient CRC cell lines (SW480 and SW620) revealed that disease stage impacts both EV uptake and drug delivery, with late-stage EVs showing reduced uptake and delivery efficiency. The protein corona, known to coat circulating EVs, was found to influence drug loading and delivery. Pre-treatment of cell line-derived EVs with plasma proteins enhanced EV uptake but reduced doxorubicin loading and subsequent delivery, particularly when using plasma from healthy volunteers. Conclusions: These findings underscore the importance of EV source and protein corona composition in optimising drug delivery strategies. Our results suggest that autologous, patient-derived EVs hold potential as a targeted drug delivery system for CRC, but highlight the need for further optimisation of EV isolation, loading methods, and understanding of how disease progression affects EV functionality. This approach could ultimately reduce systemic toxicity and improve therapeutic outcomes for CRC patients.

Pharmaceutics doi: 10.3390/pharmaceutics18060739

Authors: Olga Ozhereleva Alina Mustafaeva Anastasia Pulkina Marina Plotnikova Marina Shuklina Anna-Polina Shurygina Marina Stukova Andrej Egorov

Background/Objectives: Although influenza A viruses with partially truncated NS1 proteins are substantially attenuated and immunogenic due to enhanced innate immune activation; residual NS1-mediated antagonism of antiviral innate responses may support viral replication in the lower respiratory tract and constrain optimal immune responses. Strategies to further improve their immunogenicity and protective efficacy by incorporating immunomodulatory cytokines, such as IL-2, have been successfully explored. Methods: Here, we extended this approach to chemokine expression by engineering an NS1-truncated PR8-based virus (PR8/NS124) to express the immunomodulatory chemokine CXCL10 from the NS segment and compared it with the parental vector. Results: The recombinant NS124_SS_CXCL10 virus replicated to high titers in embryonated chicken eggs and MDCK cells. In vivo, however, CXCL10 expression reduced viral replication in mouse lungs by ~104-fold, resulting in a near-non-replicating phenotype. In contrast to the parental virus, the vector did not induce weight loss and exhibited a strongly attenuated phenotype. This effect was associated with altered innate immune signaling, including increased IRF7 expression and early induction of IFN-α responses in the lungs, together with modulation of TLR-dependent sensing pathways in the upper respiratory tract. Despite severely impaired replication, intranasal immunization induced antigen-specific T-cell responses comparable to those elicited by the parental vector. Following intraperitoneal immunization, when replication of both vectors was minimal, the CXCL10-expressing vector induced significantly higher frequencies of antigen-specific CD8+ and CD4+ effector-memory T cells. This was accompanied by enhanced antigen-specific T-cell recall responses in the lungs following intranasal challenge. Importantly, the CXCL10-expressing vector demonstrated protective efficacy comparable to that of the parental NS124 vector against heterologous H3N2 challenge while exhibiting an improved safety profile. Conclusions: These findings support the incorporation of CXCL10 as a strategy to improve the safety and T-cell immunogenicity of NS1-truncated influenza vectors.

Pharmaceutics doi: 10.3390/pharmaceutics18060738

Authors: Julia Jankowska Łukasz Szeleszczuk Dariusz Maciej Pisklak

Cancer progression is closely associated with dysregulation of apoptosis, enabling malignant cells to evade programmed cell death and develop resistance to therapy. Among the key regulators of this process, the tumor suppressor protein p53 and the Bcl-2 family of proteins play central and interconnected roles in controlling cell survival and mitochondrial integrity. In recent years, naturally occurring flavonoids have attracted considerable attention as potential modulators of these pathways due to their diverse biological activities and relatively low toxicity. This review provides a focused and integrative overview of how different subclasses of flavonoids modulate the p53–Bcl-2 signaling axis to regulate apoptosis in cancer cells. Particular emphasis is placed on the mechanistic interplay between p53 stabilization, transcriptional regulation of apoptotic targets, mitochondrial outer membrane permeabilization, and caspase activation. In contrast to previous general reviews on flavonoids and cancer, this work provides an integrated overview of evidence across multiple flavonoid subclasses and experimental cancer models, highlighting both shared and pathway-specific apoptotic responses. Experimental findings from in vitro and in vivo studies are discussed, including the effects of quercetin, kaempferol, myricetin, epigallocatechin gallate, and related compounds on cell-cycle arrest, oxidative stress, mitochondrial dysfunction, and intrinsic apoptotic signaling. Furthermore, the review examines the relationship between flavonoid chemical structure and biological activity, with particular attention to bioavailability, metabolic transformation, and strategies aimed at improving therapeutic efficacy, including structural modification and nanocarrier-based delivery systems. Despite promising preclinical findings, significant translational challenges remain, including poor pharmacokinetic properties, variability among experimental models, and limited clinical validation. Overall, flavonoids represent a promising class of bioactive compounds capable of targeting apoptosis through modulation of the p53–Bcl-2 network, and a deeper mechanistic understanding of their activity may support the development of novel targeted and combination anticancer therapies.

Pharmaceutics doi: 10.3390/pharmaceutics18060737

Authors: Syed Arman Rabbani Shrestha Sharma Mohamed El-Tanani Suman Khurana Manita Saini Monu Yadav Rakesh Kumar Yahia El-Tanani

Cancer is still one of the world’s major causes of morbidity and mortality; thus, safer and more efficient treatment approaches are required. The structural variety, multitargeted mechanisms, and generally good safety profiles of plant-derived polyphenols have made them attractive anticancer medicines. Flavonoids (like quercetin), stilbenes (like resveratrol), phenolic acids and curcuminoids (like curcumin) are major classes that have shown strong anticancer action against a variety of cancers, including prostate, colorectal and breast cancers. Through targets including PI3K/Akt, MAPK, NF-κB, and p53 signaling networks, these substances influence important molecular pathways involved in tumor initiation and development, including oxidative stress, inflammation, apoptosis, cell cycle control, angiogenesis and metastasis. The clinical translation of polyphenols is still constrained by poor bioavailability, fast metabolism, low aqueous solubility and inefficient pharmacokinetic characteristics, which lead to insufficient systemic exposure and therapeutic efficacy despite strong preclinical data. Their therapeutic applicability is further complicated by variations in absorption and possible dose-related restrictions. To overcome these limitations, the anticancer efficacy of polyphenols has been enhanced via delivery technologies like polymeric nanoparticles, lipid-based carriers, nanoemulsions and phytosome complexes, which have shown improved stability, increased bioavailability and targeted delivery to tumor tissues. This review provides a comprehensive and integrative analysis of plant-derived polyphenols by linking molecular mechanisms, pharmacokinetic limitations and emerging delivery strategies within a translational framework. By bridging these interconnected domains, this review highlights the potential of polyphenols as viable candidates in next-generation cancer therapeutics and underscores the need for well-designed clinical studies to facilitate their successful integration into oncology practice.

Pharmaceutics doi: 10.3390/pharmaceutics18060736

Authors: Sergey G. Poroshin Arkady S. Abdurashitov Gleb B. Sukhorukov Pavel I. Proshin

Background: Polymeric drug-eluting films are promising platforms for local antibacterial delivery, but their release profiles depend strongly on the permeability and morphology of the barrier layer. Here, the previously proposed concept of additively manufactured PLACE (Printed Layered Adjustable Cargo Encapsulation) coatings was extended from "single orifice"-defined release toward porosity-assisted barrier control. Two conventional water-soluble porogens, polyethylene glycol (PEG) and polyvinylpyrrolidone (PVP), were compared with poly(2-ethyl-2-oxazoline) (PEOx), a hydrophilic polymer proposed as an alternative to PEG in biomedical formulations, but whose use as a leachable porogen has received little attention. Methods: Each porogen was introduced into the upper PLGA barrier of multilayer PLACE films. The resulting films were characterized for film formation, post-hydration morphology by SEM, release of methylene blue and vancomycin, and antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). Results/Conclusions: PEG was poorly compatible with PLGA and mainly produced surface-localized defects rather than a barrier with controlled permeability suitable for prolonged delivery. PVP K17 provided sustained release at 10 wt.%, whereas 20 wt.% PVP caused burst-dominated release and stronger morphological disruption. PEOx formed developed porosity at lower loading and produced release regimes ranging from several days to approximately two weeks. Vancomycin-loaded films containing 5 wt.% PEOx enabled near-complete release over two weeks while preserving film integrity and showed pronounced early anti-MRSA activity. These results identify porogen selection as a key formulation step and support PEOx as a useful porogen for early high-output antibacterial PLACE coatings.

Pharmaceutics doi: 10.3390/pharmaceutics18060735

Authors: Alessandra Buscarini Saliha Moutaharrik Gabriele Meroni Matteo Cerea Martina Edith Coldani Anastasia Foppoli Luca Palugan Andrea Gazzaniga Piera Anna Martino Alessandra Maroni

Objectives: A customizable 3D-bioprinted core-in-shell platform was developed for time-dependent oral colon delivery of live microorganisms. The system conveyed Lacticaseibacillus paracasei as a model bacterial species within a monolithic core, which was surrounded by a swellable hydroxypropyl cellulose barrier, imparting a lag phase of programmable duration, and by an enteric outer layer, protecting the dosage form during unpredictable gastric residence. Methods: Pastes of different compositions were investigated to shape the core. Core and core-in-shell units were fabricated from digital models using a bioprinter equipped with a high-precision plunger dispenser and pressure-based thermoplastic printhead. The printed units were characterized in terms of mass, dimensions, mechanical properties and release performance using paracetamol as a reference tracer. Bacterial viability was evaluated during screening of the formulation components and after each processing step by manual counting of colony-forming units. Results: A mannitol-based formulation was selected for fabrication of the core, offering a favorable balance of printability, physico-technological properties, release behavior and ability to preserve bacterial viability. Two-layer core-in-shell systems were manufactured via a dual-printing operating mode. The desired in vitro performance was attained, with no release under acidic conditions, a lag phase in pH 6.8 fluid and a subsequent release profile comparable with that generated by the core as such. Viability studies demonstrated that compounding, core printing, shell deposition and drying did not adversely affect L. paracasei survival. Conclusions: 3D bioprinting was proved to be a versatile technique for the manufacturing of oral colon delivery systems containing probiotics or live biotherapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18060733

Authors: Miguel Marín Raúl Fuentes-Martín Baldomero Sánchez Laura Gallego-Yerga Rafael Peláez

Background/Objectives: Microtubule-targeting agents represent a pillar of cancer chemotherapy; however, their clinical utility is constrained by significant toxicity, pharmacokinetic instability, and susceptibility to multidrug resistance transporters. This study aimed to explore the impact of replacing substituted phenyl rings with a naphthalene moiety in sulfonamide-based colchicine-site ligands, with the goal of identifying new antiproliferative candidates with improved profiles. Methods: We designed, synthesized, and evaluated a library of 35 naphthalene sulfonamides bearing varied aryl groups and sulfonamide nitrogen substituents. We assessed the antiproliferative activity against multiple cancer cell lines. Mechanistic studies, including fluorescence microscopy, cell cycle analysis, and cell death assays, were performed to evaluate the effect of these compounds on microtubule polymerization dynamics and cell fate. Molecular docking and in silico pharmacokinetic profiling were carried out to support the proposed binding mode at the colchicine site and to assess drug-likeness. Results: Exclusively, compounds bearing a trimethoxyphenyl group showed antiproliferative activity in the submicromolar range, thus identifying it as a structural requirement. The most potent compound (2) reached double-digit nanomolar IC50 values (67–104 nM) across multiple cancer cell lines. Microscopy confirmed intracellular disruption of microtubule polymerization. Unlike colchicine, these compounds did not induce canonical mitotic arrest but instead triggered apoptotic cell death. In silico analyses supported binding at the colchicine site and revealed favorable predicted pharmacokinetic properties. Conclusions: The naphthalene sulfonamides described herein demonstrate potent antiproliferative activity through a distinct mechanism compared to colchicine, and their favorable in silico profiles position them as promising candidates for further development as antitumor agents.

Pharmaceutics doi: 10.3390/pharmaceutics18060734

Authors: Antonio Lopalco Oriana Boscolo Annalisa Cutrignelli Francesco Pio Cicinato Sergio Fontana Silvia Lucangioli Nunzio Denora

Background/Objectives: Ursodeoxycholic acid (UDCA) is a bile acid widely used for the treatment of cholestatic liver diseases; however, its poor aqueous solubility represents a major limitation for the development of oral liquid formulations, particularly in pediatric patients requiring accurate and flexible dosing. This study aimed to develop and characterize a fully solubilized extemporaneous UDCA oral formulation using the ready-to-use vehicle Wagner, with particular emphasis on the role of hydroxypropyl-β-cyclodextrin (HP-β-CD) as a solubilizing excipient. Methods: Phase-solubility studies, Job’s plot analysis, and 1H NMR spectroscopy were performed to investigate the host–guest interaction between UDCA and HP-β-CD, confirming the formation of a stable 1:1 inclusion complex responsible for a marked increase in drug solubility. The aqueous solubility of UDCA increased from approximately 0.02 mg/mL in water to 31 ± 1 mg/mL in the Wagner base containing HP-β-CD, compared to ~10 mg/mL in the corresponding cyclodextrin-free vehicle. Chemical stability was evaluated using an HPLC method adapted from the European Pharmacopoeia, employing dual detection (refractive index and photodiode array detector) to ensure specificity and stability-indicating capability. Results: The UDCA solution (20 mg/mL) remained chemically stable for at least 4 months under refrigerated (4–8 °C) and room temperature (25 °C) conditions, with only moderate degradation observed at 40 °C. Physical stability studies confirmed the absence of precipitation, phase separation, or significant pH variations under all storage conditions. Conclusions: Wagner-based formulation enabled the development of a stable and homogeneous UDCA oral solution, providing a complementary formulation strategy to conventional suspension-based preparations. This approach represents a robust and patient-oriented strategy for extemporaneous compounding, particularly suitable for pediatric use.

Pharmaceutics doi: 10.3390/pharmaceutics18060732

Authors: Oindrila Palit Ankita Das Supriya Bharti Eirinaios I. Vrettos Sankarprasad Bhuniya

Efficient delivery systems are essential for transporting chemotherapeutic agents to target sites, enhancing cellular uptake and reducing off-target side effects. Peptides, owing to their intrinsic biocompatibility and structural tunability, have emerged as promising carriers for delivering labile chemotherapeutics and improving pharmacokinetics and therapeutic outcomes. Along these lines, a wide variety of peptide-based delivery strategies have been developed to achieve desirable pharmaceutical properties for anticancer agents. Particularly, stimuli-responsive peptide-based nanocarriers have attracted high levels of attention due to their ability to exploit overexpressed or tumor-specific stimuli, enabling selective disassembly and controlled drug release within cancer cells. In this review, we highlight recent advances in the development of stimuli-responsive peptide nanocarriers and their applications in anticancer therapy, and discuss key challenges and future directions toward their clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18060731

Authors: Hyung-Ju Seo Zhuoning Liang Eui-Hyeon Kim Kwang-Hyeon Liu

Background/Objectives: Atrasentan is a selective endothelin A receptor antagonist (SERA) developed as a potential therapy for chronic renal diseases, including diabetic nephropathy and immunoglobulin A nephropathy. Despite this potential, understanding its metabolic bioactivation is essential for assessing the risks of drug-induced liver injury (DILI). However, the metabolic profile of atrasentan remains poorly characterized, and the mechanisms underlying its potential hepatotoxicity remain underexplored. Therefore, this study aims to investigate the metabolic pathways of atrasentan in human liver microsomes (HLMs) in the presence of nicotinamide adenine dinucleotide phosphate (NADP+), uridine diphosphate glucuronic acid (UDPGA), or glutathione (GSH). Methods: A liquid chromatography–high resolution mass spectrometry (LC-HRMS) coupled with a feature-based molecular networking approach was used to characterize metabolites. Characterization of the major metabolites was achieved through cytochrome P450 (P450) phenotyping with human recombinant P450 isoforms. Results: A total of eighteen metabolites were characterized through phase I and II metabolic reactions, including demethylenation, N-dealkylation, O-demethylation, hydroxylation, dehydrogenation, and glucuronidation. Atrasentan acyl glucuronide (M8) was confirmed as the predominant metabolite, and we also putatively annotated a catechol intermediate (M5) and its corresponding GSH conjugate (M15). Characterizing the GSH conjugate (M15) indicates that catechol intermediate (M5) can be further oxidized to a reactive ortho-quinone intermediate, which is subsequently trapped by GSH, suggesting the potential for a bioactivation mechanism. Reaction phenotyping demonstrated that the formation of M5 is catalyzed almost exclusively by the CYP3A subfamily. However, its direct translation to in vivo oxidative stress or covalent protein binding requires further studies. Conclusions: These findings demonstrate that feature-based molecular networking is a valuable strategy for metabolite characterization, underscoring the urgent need for further in vivo metabolism studies to definitively assess hepatotoxic risks associated with these reactive metabolites.

Pharmaceutics doi: 10.3390/pharmaceutics18060730

Authors: Eva Costanzi Luca Fontana Francesca Giroldo Silvia Coco

Neuroinflammation plays a central role in the onset and progression of neurodegenerative disorders. Several disease-modifying therapies have been developed to target neuroinflammatory pathways in specific disorders. However, their ability to stop disease progression or restore neuronal and mitochondrial homeostasis remains limited. This is still a major unmet clinical need. In this context, mesenchymal stromal cell (MSC)-derived Extracellular Vesicles (EVs) have emerged as a promising cell-free therapeutic strategy due to their ability to modulate immune responses and promote neuroprotection through the delivery of bioactive cargo. Recent evidence has identified a distinct subset of EVs, known as mitochondrial EVs (mito-EVs), which carry mitochondrial DNA, proteins, and functional components. These vesicles may uniquely influence cellular bioenergetics, redox balance, and neuroinflammatory signaling, offering additional therapeutic potential compared to conventional MSC-EVs. This review summarizes the role of MSC-derived EVs in neuroinflammatory disorders, with a particular focus on mito-EVs. It also discusses preconditioning strategies to enhance EV efficacy, including hypoxic, inflammatory, pharmacological priming and genetic engineering approaches. Finally, we critically evaluate current preclinical evidence regarding the treatment of major neurodegenerative disorders, including Alzheimer’s disease, Parkinson’s disease, Multiple Sclerosis, and Amyotrophic Lateral Sclerosis, as well as Traumatic Injury, highlighting the key challenges for clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18060729

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

Cutaneous infections and chronic inflammatory wounds remain difficult to treat because antimicrobial resistance, polymicrobial biofilms, excessive protease activity, oxidative stress, and impaired barrier repair collectively reduce the effectiveness of conventional topical therapies. Plant-derived antimicrobial peptides (AMPs) and peptide-associated bioactives offer antimicrobial, antibiofilm, immunomodulatory, and tissue reparative potential; however, their clinical translation is limited by proteolytic instability, poor stratum corneum penetration, short cutaneous residence time, formulation variability, cytotoxicity risks and limited human evidence. The key research gap is the lack of an integrated translational framework linking plant-derived peptide bioactivity with polymer engineering, advanced delivery systems, skin microenvironment biology, manufacturability, and regulatory feasibility. This review aims to critically evaluate the design principles, therapeutic mechanisms, delivery platforms, and translational barriers of plant-based peptide–polymer therapeutics for cutaneous infection and inflammation. We summarize major classes of plant-derived antimicrobial peptides, including defensins, cyclotides, thionins, hevein-like peptides, snakins, lipid transfer proteins, and knottin-type scaffolds, and examine engineering strategies such as self-assembly, aromatic N-capping, PEGylation, lipidation, dendritic architectures, and stimuli-responsive conjugation. We further discuss topical matrices, nanocarriers, liposomes, electrospun fibers, and surface-tethered biomaterials as delivery platforms for improving peptide stability, local retention, and controlled release. Finally, we identify key translational bottlenecks, including selectivity, toxicity, scalability, batch reproducibility, regulatory classification, and insufficient clinical validation. Mechanism-driven peptide optimization, quality-by-design manufacturing, standardized preclinical models, and controlled clinical trials will be essential for advancing these systems toward safe and effective dermatological therapies.

Pharmaceutics doi: 10.3390/pharmaceutics18060728

Authors: Fatin Jannus Hilario Ramírez-Rodrigo

Background/Objectives: Traditional protein contact analysis often fails to distinguish between local, sequence-driven motifs and global, tertiary scaffolding, which ensures structural determinism. While deep-learning models do not fully elucidate the ‘why’, they do reveal the underlying directional rules of the stability landscape. In this study, we analyzed 475 non-redundant Protein Data Bank (PDB) structures categorized into SCOP classes (all-α, all-β, α/β, α+β) of the hydrolase superfamily. Methods: To isolate the structural anchors of the global fold, we applied a sequence separation filter of ∣i − j∣ ≥ 6 and a precise spatial cutoff of 3–5 Å between Cα-only to construct asymmetric 20 × 20 frequency matrices, both raw and normalized, then present the former using a violin diagram. We developed a Pearson Correlation (PC) framework to analyze these matrices, providing high correlation when considered as vectors and giving the directionality (N-to-C vs. C-to-N) in protein folding when considered as matrices. Results: Our results reveal a hierarchical organization of tertiary determinism. Initial visualization of Residue–Residue Contact Frequency Matrices (RRCFMs), Z-score normalization (NRRCFM), and violin plots reveal the Universal Structural Grammar (USG) of interaction. Furthermore, a near-perfect PC (r = 0.99) as determined via inter-class Z-score correlation and inter-class PC demonstrates shared statistical interaction laws. In addition, PC Stage 1 (intra-class) analysis identified high symmetry, with around 80% of contacts exhibiting a very strong to strong positive correlations, while PC Stage 2 (inter-class) analysis demonstrated that around 50% of contacts exhibited very strong to strong positive correlations. Finally, we identified universal druggable pockets for drug discovery. Conclusions: This powerful mathematical framework provides a robust analytical tool for structure-based drug design.

Pharmaceutics doi: 10.3390/pharmaceutics18060727

Authors: Eva Carolina Arrua Cintia Briones Nieva Santiago Nicolás Campos Andrea Paola Rivas Marquina Giselle R. Bedogni Claudia Llanos Alicia Graciela Cid Mercedes Villegas Elio Emilio Gonzo Claudio Javier Salomon José María Bermúdez

Background/Objectives: Lipid-based nanocarriers have emerged as promising systems for improving the delivery of poorly soluble drugs by enhancing stability, bioavailability, and controlled release. This work aimed to formulate solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) containing ciprofloxacin (CIP) using solvent-free procedures. Methods: The systems were extensively characterized using dynamic light scattering (DLS), transmission electron microscopy (TEM), and atomic force microscopy (AFM) to study the nanoparticles in the solid state. Furthermore, in vitro drug release was evaluated, and mathematical modeling was applied to analyze the resulting release kinetics. Additionally, storage stability was assessed at 4 °C and 25 °C over a period of 8 months. Results: The results indicated that SLN with an average size of ~50 nm (SLN 50) and NLC with mean diameters of ~25, 50, and 100 nm (NLC 25, NLC 50 and NLC 100 respectively) were successfully prepared. DLS measurements showed narrow particle size distributions (PdI ≤ 0.2) and negative zeta potentials ranging from −3.7 to −7.7 mV. Encapsulation efficiencies were remarkably high for most systems, reaching ~98% for SLN 50, NLC 50, and NLC 100, while the smallest formulation (NLC 25) showed a lower efficiency (~80%). Both TEM and AFM confirmed the formation of spherical nanoscale structures consistent with the sizes determined by DLS. Release studies revealed a strong influence of particle size on kinetics: NLC 25 exhibited rapid release (~95% within 30 min), whereas NLC 100 showed a sustained profile (<20% after 6 h). Dissolution profiles were accurately described by the Lumped-Gonzo kinetic model (R2 > 0.98), enabling estimation of dissolution efficiency. Conclusions: These findings confirm that lipid-based nanocarriers can be engineered to precisely control CIP release.

Pharmaceutics doi: 10.3390/pharmaceutics18060726

Authors: Xingjian Liu Wei Zhang Guanyong Deng Haozhe Yu Shilin Tian Jiahui Liu Wenzeng Hu Tianyu Pan Lihong Fan

Background: Conventional and refractory wounds frequently remain in a prolonged inflammatory phase associated with excessive reactive oxygen species (ROS) accumulation and disruption of endogenous electrical cues. Methods: A multifunctional nanocomposite hydrogel was fabricated via an amidation condensation reaction, utilizing 3-amino-4-methoxybenzoic acid (AMB)-modified carboxymethyl chitosan (PAMB-CMCS) and decellularized extracellular matrix (dECM) as macromolecular networks, integrated with Astragaloside IV-Loaded Polydopamine/Zeolitic Imidazolate Framework-8 (AS@PDA/ZIF-8) nanoparticles. Results: The hydrogel provided a biomechanically supportive scaffold with compressive strength of 27.24 ± 1.9 kPa and breaking strength of 28.2 ± 2.8 kPa and exhibited electrical conductivity of 29.84 mS/cm, ROS-scavenging activity, and near-infrared (NIR)-responsive photothermal behavior reaching 62.55 °C. The integrated PDA@ZIF-8 nanoplatform further contributed to antibacterial performance and localized AS release, thereby improving the wound microenvironment and accelerating full-thickness cutaneous defect repair. Conclusions: This macromolecule-based composite hydrogel offers a promising therapeutic strategy for complex wound management.

Pharmaceutics doi: 10.3390/pharmaceutics18060725

Authors: Nan Wu Chaocheng Wu Yating Du Zhuhua Chan Runying Zeng

Background/Objectives: Neoagarotetraose (NA4), a marine-derived tetrasaccharide, holds promise as an anti-inflammatory and antioxidant agent; however, its oral bioavailability and systemic exposure mechanisms require elucidation. Methods: This study characterizes the biopharmaceutical profile of NA4 after oral and intravenous administration using a validated near-infrared fluorescence method based on covalent conjugation with Cy7. Results: Following oral gavage (200 mg/kg), NA4-Cy7 was rapidly absorbed (Tmax: 1.0 h; Cmax: 35.6 mg/L), with prolonged systemic exposure (mean residence time: 13.1 h) and an elimination half-life of 8.9 h. Intravenous administration (25 mg/kg) revealed a low volume of distribution at steady state (Vss: 0.0132 L/kg) and a shorter MRT (4.3 h). Tissue distribution at 24 h showed preferential accumulation in the kidney, liver, and lung, with direct visualization of intact NA4 crossing the intestinal epithelium. Conclusions: These findings demonstrate that fluorescently labeled NA4-Cy7 can cross the intestinal epithelial barrier and reach systemic circulation, supporting its potential as an orally active agent with organ-specific targeting properties.

Pharmaceutics doi: 10.3390/pharmaceutics18060724

Authors: Natália Yukari Kashiwaqui Helena Tiemi Suzukawa Briani Gisele Bigotto Maria Luiza Francisconi Lubanco Thomé Danielle Lazarin Bidoia Sueli Fumie Yamada-Ogatta Ricardo Sérgio Couto de Almeida Audrey Alesandra Stinghen Garcia Lonni Mirian Sumini Renata Katsuko Takayama Kobayashi Gerson Nakazato

Background/Objectives: Burn injuries represent a global public health concern, accounting for approximately 265,000 deaths annually and often leading to severe infections. With the increasing prevalence of multidrug-resistant (MDR) bacteria, innovative therapeutic strategies such as nanoparticle-based topical formulations have gained attention. This study proposed the development of a hydrogel for burn treatment containing biogenic silver nanoparticles (BioAgNPs), hyaluronic acid (HA), and allantoin (AL). Methods: BioAgNPs were previously characterized by transmission electron microscopy (TEM) and incorporated into a hydrogel containing HA and AL, which was physicochemically characterized by pH, spreadability, and energy-dispersive X-ray spectroscopy (EDX). Antibacterial activity was evaluated by broth microdilution, agar diffusion, and time–kill assays against standard and MDR bacterial strains. Cytotoxicity was assessed using the MTT assay in L929 cells, and wound-healing potential was investigated through an in vitro scratch assay to evaluate cell migration and proliferation. Results: BioAgNPs exhibited antibacterial activity against reference strains and MDR isolates, determining the minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC). HA and AL were non-toxic, while BioAgNPs demonstrated low cytotoxic activity. Although HA and AL did not exhibit antibacterial properties, they promoted cell migration and proliferation. The formulation exhibited physicochemical and pharmaceutical stability, showing suitable properties for topical use, and presented significant antimicrobial action, with bacterial elimination occurring within 2 h of contact, except for S. aureus. Conclusions: Thus, the hydrogel presents a promising alternative for the topical treatment of infected burns, with potential application in combating multidrug-resistant bacteria, being able to eliminate MDR Acinetobacter baumannii.

Pharmaceutics doi: 10.3390/pharmaceutics18060723

Authors: Yasmim Vilarim Barbosa Maria Elaine Cristina Araruna Maria Lorenna Pessoa Fonsêca Francisco José Batista de Lima Júnior Cassiano Francisco Weege Nonaka Paulo César Dantas da Silva José Elizandro Batista de Oliveira Bruna Larissa Barbosa de Lira Thássia Borges Costa Vanda Lucia dos Santos

Background/Objectives: Inflammatory bowel diseases (IBD) are conditions of the gastrointestinal tract with treatments linked to side effects and relapses. Spondias mombin L. is a species with anti-inflammatory action, but there is little information in the literature about its application in the treatment of IBD. The aim of this study was to evaluate the intestinal anti-inflammatory activity of hydroalcoholic extract of Spondias mombin L. (HESm) in a rat model of ulcerative colitis (UC). Methods: Hydroalcoholic extract was obtained using the turboextraction technique, followed by identification of its major components using ultra-high performance liquid chromatography (UFLC). Intestinal anti-inflammatory activity was evaluated in an acute model of UC induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). Groups received a vehicle, prednisolone (2 mg/kg), or HESm (125, 250 or 500 mg/kg) before and after UC induction. Ulcerated area, score, and intestinal weight/length ratio were analyzed. Histopathological analysis and the extract’s effect on the contractility of the intestinal segment were carried out. Results: UFLC identified the presence of quercetin, a flavonoid widely cited for the species. At doses of 125, 250, and 500 mg/kg, the extract reduced areas of injury by 86.82, 92.67, and 85.06%, respectively, compared to the control, in addition to reducing scores and weight/length ratio of the colons. Histopathological analysis confirmed the results. In contractility, the extract at the highest concentration tested reduced the response of the muscarinic agonist carbamylcholine to 53.7 ± 4.2% of control contraction. Conclusions: Results demonstrate the species’ ability to reduce injuries caused by colitis, suggesting its potential to contribute to the clinical management of IBD in the future.

Pharmaceutics doi: 10.3390/pharmaceutics18060722

Authors: Bárbara Pinto Joaquim Teixeira de Avelar Júnior Edleusa Marques Lima Lívia Ramos Santiago Rosy Iara Maciel de Azambuja Ribeiro Renata Toscano Simões Cristina Moraes Junta Rachel Carolina Souza Fagundes Ana Clara Costa Velozo Hassan Bousbaa Miriam Teresa Paz Lopes Juliana Carvalho-Tavares Elaine Maria de Souza-Fagundes Maria Elena de Lima

Cancer, a longstanding global challenge, remains a leading cause of death, prompting an urgent search for effective treatments. Conventional therapies, while prevalent, often cause adverse effects due to their lack of specificity. This review explores an innovative approach, focusing on animal toxins as a rich source of bioactive compounds which have demonstrated efficacy against cancer cells. Peptides from various species, including scorpions, snakes, bees, spiders, and frogs, show promising antiproliferative and cytotoxic effects. Emphasizing the most prevalent types of cancer, this review outlines the discovery and development stages of potential anticancer drugs derived from toxin peptides. The comprehensive overview includes in vitro and in vivo assessments of their anticancer activity and toxicity. This pioneering exploration extends to different tumors, offering valuable insights into mechanisms of action and potential therapeutic applications. The findings highlight a paradigm shift in cancer research, showcasing the potential of toxin-derived compounds for developing targeted and efficient cancer therapies with reduced side effects.

Pharmaceutics doi: 10.3390/pharmaceutics18060721

Authors: Daniel Zakowiecki Peter Edinger Michael Wagner Tobias Hess Dariusz Lipiak Krzysztof Cal

Background: An increasing number of commercially available drug substances and bioactive ingredients are characterized by poor flowability. Inadequate flow properties may lead to material blockage during transport within production lines, as well as the formation of air voids within the bulk. Such phenomena can disrupt the technological process and may even result in batches that fail to meet quality requirements. Therefore, ensuring adequate powder flow is of utmost importance in the manufacture of health-related products. Methods: Binary mixtures were prepared using one of four model substances (ibuprofen, metamizole sodium, mefenamic acid, or sunflower lecithin) combined with a glidant (colloidal silica, precipitated silica, or tricalcium phosphate). The glidant content ranged from 0.5 to 10.0% w/w depending on the model substance, and mixing was carried out for 5–30 min. The resulting binary mixtures were evaluated for flow properties using the angle of repose method, and in selected cases, bulk density was also determined. Results/Conclusions: The study demonstrated that powder flow improvement depended not only on the glidant but primarily on the properties of the host material (particle size, shape, and bulk density). Coarser powders such as ibuprofen responded well to low glidant levels, although excessive silicon dioxide caused oversilication. Metamizole sodium showed progressively better flow with increasing particle size and density, and tribasic calcium phosphate further improved performance, particularly with longer mixing times. Very fine or cohesive powders, such as mefenamic acid and sunflower lecithin, showed limited response to silica-based glidants, whereas tribasic calcium phosphate proved more effective and additionally increased bulk density. Overall, no universal glidant strategy was identified; effective flow enhancement requires a tailored approach based on specific powder characteristics.

Pharmaceutics doi: 10.3390/pharmaceutics18060720

Authors: Antonio Giuseppucci Jianing Chen George Hussey Héctor Capella-Monsonís

Background: Matrix-bound nanovesicles (MBV) are extracellular vesicles (EVs) that are embedded within the extracellular matrix (ECM), and they have shown immunomodulatory effects in various cell types. The THP-1 cell line is often used to study monocyte and macrophage functions due to its easy culture potential and relatively simple conditioning into different macrophage phenotypes, but the optimal culturing conditions that allow MBV immunomodulation have not been established. Methods: In this study, we evaluated different culturing and differentiation conditions of THP-1 cells in which MBVs showed immunomodulatory effects. We also studied the effect of MBVs on relevant inflammation pathways (NF-κB and ERK 1/2). Results: Quantification of inflammatory cytokine IL-6 indicated modulation effects by MBVs in the majority of the conditions, but TNF-α showed very limited modulation. ERK1/p44 phosphorylation was significantly increased in MBV groups, but NF-κB protein p65 expression was unaffected. When compared to serum EVs, vesicle uptake by THP-1 cells remained low after 24 h. Multispectral flow cytometry analysis of THP-1 cells exposed to MBV and serum EVs showed internalization of lipids, proteins and RNA within the cells in higher cell proportions, but colocalization of the different vesicle components was not observed. Conclusions: Overall, this study provided insights into MBV immunomodulatory effects on THP-1 cells and compared the effects of MBV and serum EVs. Slight differences in modulation were observed between both EV sources, pointing to cargo differences that need further investigation.

Pharmaceutics doi: 10.3390/pharmaceutics18060719

Authors: Eslam Ramadan Nooh Mdrmah Martin Deák Norbert Varga Edit Csapó Tamás Sovány Katalin Kristó

Background: Oral protein delivery is a major challenge in the field of pharmaceutical technology due to poor stability and limited permeability through intestinal barriers. Buccal delivery is a promising alternative with less restricting physiological conditions; however, low protein permeability is still a limiting factor. Multiple nanocarriers have been proposed to improve buccal protein delivery with lipid–polymer hybrid nanoparticles (LPHNs) combining the advantages of both polymeric and lipid-based systems. However, these conventional carriers rely on passive protein protection and lack adaptive release mechanisms. Objectives: This work aimed to develop and systematically optimize an ionic strength-responsive LPHN system that can minimize protein release in buccal ionic conditions while offering a triggered release in plasma after absorption. Methods: LPHNs were prepared by a two-step approach where polymeric cores of Eudragit-L100 were prepared by electrostatic complexation with Lysozyme (LYZ) followed by lipid shell formation by the ethanol injection method. Systematic optimization was performed using two-level factorial and central composite designs. Moreover, the ionic strength responsiveness and in vitro LYZ release were investigated in different ionic strength media. Results: The final optimized formulations, LPHNs and sodium deoxycholate-containing LPHNs (NaDC-LPHNs), exhibited a particle size of 257.2 ± 1.5 nm and 246 ± 5.7 nm, encapsulation efficiency of 69.89 ± 0.22% and 68.14 ± 0.16%, and high drug loading efficiency of 24.11 ± 0.06% and 23.65 ± 0.04%, respectively. Moreover, both formulations showed minimal protein release at low ionic strength (buccal-like) conditions while demonstrating a triggered release at higher ionic strength (plasma-like) conditions. Conclusions: The developed system may provide a promising smart strategy to improve buccal protein delivery by enhancing buccal protection and improving systemic delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18060718

Authors: Kai-Juan Cao Long Fu Yu-Chen Sun Jian Meng Qin Huang De-Cheng Deng Hai-Tang Hu Zhi-Hui Han Gang Guo Xue Zhou Xiao-Yan Chen

Background/Objectives: Zastaprazan (JP-1366) is a novel potassium-competitive acid blocker (P-CAB) used for the treatment of gastroesophageal reflux disease (GERD). To date, its metabolic pathways and metabolism-related drug–drug interactions (DDIs) in humans remain incompletely elucidated. This study aimed to determine the relative contributions (fm) of cytochrome P450 isoforms to JP-1366 elimination and assess its DDI potential. Methods/Results: In vitro metabolic studies using human liver microsomes (HLMs) revealed that JP-1366 was first metabolized to M1, which subsequently underwent further oxidation, glucuronidation, and N-dealkylation. Mono-oxidation was estimated to contribute more than 46% to the overall metabolic clearance of JP-1366. Reaction phenotyping identified CYP3A as the major enzyme (fm = 96.1%), followed by CYP1A2 (1.49%) and CYP2C9 (2.41%). By integrating in vitro data, clinical pharmacokinetic data and clarithromycin coadministration DDI data, a physiologically based pharmacokinetic (PBPK) model was developed and validated. Simulations predicted significant DDIs with strong CYP3A inhibitor (ketoconazole), with AUC ratios of 3.80. Moderate inhibitors (fluconazole and fluvoxamine) caused mild increases (AUC ratios: 1.14–1.74). Conversely, strong and moderate CYP3A inducers, rifampicin and efavirenz, produced pronounced DDIs, with AUC ratios of 0.22 and 0.50, respectively. Furthermore, simulations predicted that although JP-1366 functions as a CYP enzyme inhibitor, it would not cause clinically meaningful changes in the plasma exposure of corresponding CYP substrate drugs; however, potential interactions with CYP3A substrates still warranted consideration. Conclusions: JP-1366 is predominantly cleared via a CYP3A-dominated metabolic pathway. The PBPK simulations suggest that JP-1366 may be a moderately sensitive CYP3A substrate and a moderate inhibitor of sensitive CYP3A substrates, while its perpetrator DDI risk toward other major CYP pathways appears limited. These findings support caution or monitoring when JP-1366 is co-administered with strong CYP3A modulators or sensitive CYP3A substrates.

Pharmaceutics doi: 10.3390/pharmaceutics18060717

Authors: Eman Marzouk Ayman Elbehiry

Carbapenem-resistant Enterobacterales (CRE) are a major therapeutic challenge because of limited treatment options and high mortality. Despite advances in resistance-targeted therapies and pharmacokinetic (PK) optimization, treatment failure remains common. This review examines how resistance mechanisms and antibiotic exposure at the infection site jointly influence therapeutic outcomes in CRE infections. A mechanistic synthesis of evidence on carbapenem PKs, bacterial resistance, and nanoparticle (NP)-based delivery systems was performed. Based on this analysis, we propose the Pre-Target Interception Model (PTIM), which describes treatment failure as the progressive loss of active antibiotic before reaching penicillin-binding proteins. Unlike conventional approaches that focus primarily on resistance determinants or drug delivery platforms, PTIM emphasizes the factors that limit effective antibiotic exposure within infected tissues. Within this framework, nanocarrier systems are assessed according to their ability to protect antibiotics, enhance tissue penetration, and improve retention under conditions of enzymatic degradation, membrane restriction, efflux activity, and biofilm-associated diffusion barriers. However, clinical translation remains limited by manufacturing challenges, variability in NP performance, and the lack of validation in CRE-specific settings. Future progress will require quantitative measurement of antibiotic exposure at infection sites, standardized evaluation of nanocarrier performance, and validation in clinically relevant models. PTIM provides a framework for the rational development of nanomedicines designed to improve antibiotic delivery in CRE infections.

Pharmaceutics doi: 10.3390/pharmaceutics18060716

Authors: Ayman Elbehiry Adil Abalkhail Fahad A. Alhumaydhi Eman Marzouk

Carbapenem-resistant Acinetobacter baumannii (CRAB) is considered a persistent clinical problem characterized by high mortality and restricted therapeutic options. The current antimicrobial regimen focuses on active bacteria without taking into account physiological states that influence the treatment response. Biofilm formation, metabolic changes, efflux activity, and membrane remodeling reduce antibiotic activity at infection sites and help bacteria survive despite in vitro susceptibility. Clinical performance is also compromised by inadequate tissue penetration, toxicity, and inconsistent pharmacokinetics, which reduce the ability to maintain effective antimicrobial activity at the target site. Therefore, a new strategy is needed that considers how bacteria behave during infection. Nanotherapeutic systems can optimize antimicrobial delivery by changing drug distribution and enabling sustained antimicrobial release within infected tissues. These properties can improve antimicrobial distribution within biofilms and structurally restricted infection sites. This review proposes a phenotype-guided approach linking dominant bacterial phenotypes with targeted nanotherapeutic intervention. Advances in nanoscale diagnostics and computational analysis allow earlier identification and more precise characterization of resistance features, so treatment decisions reflect the current state of infection. When integrated with nanotechnology, this information supports treatment approaches that adapt to changes in bacterial behavior over time. Extending this concept to host-directed and microbiome-informed interventions provides additional control by addressing factors that sustain infection beyond the pathogen. These elements create an integrated system that connects detection, analysis, and treatment, allowing therapy to match the biological conditions of infection for more precise CRAB management.

Pharmaceutics doi: 10.3390/pharmaceutics18060715

Authors: Ting Zou Lanlan Ni Keqiang Xu Yi Chang

Background/Objectives: The rapid emergence of multidrug-resistant (MDR) bacteria has increased the demand for non-antibiotic antibacterial strategies. Although photothermal therapy (PTT) and photodynamic therapy (PDT) are promising alternatives, each modality alone may show limited antibacterial efficacy. This study aimed to construct a flexible dual-light-responsive nanofibrous membrane integrating PTT and PDT for improved in vitro antibacterial activity against MDR bacteria. Methods: A silica nanofibrous membrane (SNF) was prepared by electrospinning followed by calcination. An Fe-doped sulfonated TpPa covalent organic framework (SCOF-Fe) was then grown in situ on the SNF surface via an interfacial diffusion strategy to obtain SNF@SCOF-Fe. The membrane was characterized in terms of morphology, structure, optical absorption, photothermal performance, Fe loading, Fe leaching, and reactive oxygen species (ROS) generation. In vitro antibacterial activity against supplier-reported MDR Escherichia coli (E. coli) and methicillin-resistant Staphylococcus aureus (MRSA) was evaluated under 420 nm, 808 nm, and dual-light (420 + 808 nm) irradiation. Results: Fe doping broadened the optical absorption of the COF-functionalized membrane into the near-infrared region and improved its photothermal response. Under dual-light irradiation, SNF@SCOF-Fe generated singlet oxygen, superoxide radicals, and hydroxyl radicals, together with a greater temperature increase than the undoped membrane. Within 15 min, SNF@SCOF-Fe achieved antibacterial rates of 99.29% against E. coli and 99.62% against MRSA. In addition, controlled dual-light cytocompatibility testing yielded 78.76% viability in L929 fibroblasts and 82.86% viability in MC38 murine colon carcinoma cells after SNF@SCOF-Fe treatment. Conclusions: SNF@SCOF-Fe combines dual-light-triggered photothermal heating and ROS generation within a flexible nanofibrous membrane and demonstrated effective in vitro antibacterial activity against two representative resistant bacteria. These findings support further investigation of SNF@SCOF-Fe as a light-responsive antibacterial membrane in relevant in vitro and in vivo models.

Pharmaceutics doi: 10.3390/pharmaceutics18060714

Authors: María Elena Sánchez-Mendoza Jesús Arrieta Yaraset López-Lorenzo Gisela Gutiérrez-Iglesias Osmar Antonio Jaramillo-Morales Josué Vidal Espinosa-Juárez

Background/Objectives: Heliotropium indicum (H. indicum) is a medicinal plant traditionally used for conditions associated with inflammation, but its active antinociceptive constituents remain poorly defined. This study evaluated the antinociceptive activity of the aerial parts of H. indicum through a bioassay-guided approach and explored the mechanism of action of the active compound isolated in the formalin test. Methods: Three extracts of H. indicum (hexane, dichloromethane, and methanol) were evaluated in male Swiss albino CD-1 mice using the formalin test. The most active extract was fractionated, and its major fractions were screened for antinociceptive activity. Based on the active fraction and previous phytochemical data, (E)-ethyl-12-cyclohexyl-4,5-dihydroxydodec-2-enoate (ECDE) was selected for further pharmacological evaluation in the same model. Antagonist pretreatments were used to investigate the involvement of opioid, serotonergic, gamma-aminobutyric acid (GABAA), and Nitric Oxide (NO)–soluble Guanylyl Cyclase (sGC) pathways. Results: The three extracts reduced nociceptive behavior, mainly during phase II of the formalin test, whereas the dichloromethane extract showed the broadest activity profile and was selected for fractionation. The six fractions significantly reduced phase II nociception, and fraction F5 was selected for purification. ECDE produced a clear dose-dependent antinociceptive effect in phase II, with minimal effect on phase I, and its efficacy was compared with that of ketorolac, a standard antinociceptive drug. Dose–response analysis estimated a DE50 of 0.76 mg/kg for ECDE. Pretreatment with N-nitro-L-arginine methyl ester (L-NAME) and [1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one (ODQ) significantly attenuated the effect of ECDE, whereas naloxone, methiothepin, and bicuculline did not. Conclusions: ECDE was identified in H. indicum as one of the compounds contributing to this effect. Its activity appears to be directed mainly toward inflammatory nociception and to depend, at least in part, on the NO–sGC pathway.

Pharmaceutics doi: 10.3390/pharmaceutics18060713

Authors: Ágnes Rusznyák Péter Magyar Virág Dajka Alexandra Gyöngyösi István Lekli György Vámosi Milo Malanga Éva Fenyvesi Lajos Szente Judit Váradi Ildikó Bácskay Eszter Puhl Ferenc Fenyvesi

Background/Objectives: Intracellular delivery of RNA molecules is challenging. To solve this problem, many carrier systems are available, which are based on liposomes or polymers. Cyclodextrins are widely used excipients to increase the solubility of small molecules, but their polymer derivatives are able to deliver macromolecules. In the present study, we aimed to investigate and compare the siRNA and mRNA carrying capacity of a cationic quaternary ammonium β-cyclodextrin polymer (QABCDPS) and polyethylenimine (PEI). Methods: Cytotoxicity of the polymers was tested by the MTT method. Polyplexes were formulated with different nitrogen/phosphate ratios (NP), and their physicochemical properties were examined using dynamic light scattering and zeta potential measurements. Cellular internalization and intracellular effects of the polyplexes were investigated by confocal microscopy and flow cytometry. Results: QABCDPS exhibited lower toxicity compared to PEI, effectively binding both siRNA and mRNA and delivering them into vesicles in the cytoplasm, but showing different internalization patterns. Polyplexes formed with PEI showed stronger biological effect than those with QABCDPS, which can be attributed to the strength of interactions facilitated by the polymers. Conclusions: In summary, QABCDPS is a low-toxicity carrier that shows some promise for mRNA delivery but is ineffective for siRNA silencing under the tested conditions and requires further structural optimization.

Pharmaceutics doi: 10.3390/pharmaceutics18060712

Authors: Mingliang Su Zhiwei Liang Long Chen Taoyu Wang Shabatula Aisika Yanbin Peng Yonghui Wu Huahe Zhu Lixin Wang

Camptothecin-class compounds are anticancer active ingredients extracted from the Chinese unique medicinal plant, Camptotheca acuminata. They have been widely applied in the treatment of various cancers due to their high efficacy and broad-spectrum anti-cancer properties. This article summarizes the latest research progress over the past decade on various types of nanocarriers for camptothecin drugs. We discuss the nanodrug delivery systems for camptothecin compounds in four perspectives: passive targeting nanoparticles, active targeting nanoparticles, tumor microenvironment-responsive nanoparticles and exogenous stimulus-responsive nanoparticles. We also elaborate on the advantages of delivery nanoparticles, in vivo release characteristics and antitumor therapeutic effects. The purpose of this article is to provide a theoretical basis and innovative perspectives for the clinical application of the camptothecin drugs and the development of new pharmaceuticals.

Pharmaceutics doi: 10.3390/pharmaceutics18060711

Authors: Francisca Acevedo Manuel Azocar Eulàlia Sans-Serramitjana Jeyson Hermosilla Felipe Gálvez-Jirón Denisse Bravo Dayaimi Gonzalez Gabriela Guajardo Cristóbal Guajardo Rodrigo Navia

Background/Objectives: Wound infections represent a major clinical challenge due to their polymicrobial nature, biofilm formation, and increasing antimicrobial resistance, which compromise conventional treatments. This study aimed to develop and evaluate ligand-stabilized silver nanoparticles (AgNPs) with improved antimicrobial activity and cytocompatibility, and to investigate their incorporation into electrospun nanofibers for wound management. Methods: Four AgNP formulations stabilized with citrate, cysteine, ketorolac, and diclofenac were synthesized via chemical reduction. Physicochemical characterization included surface plasmon resonance and zeta potential measurements. Antimicrobial activity was assessed through minimum inhibitory concentration (MIC) and bactericidal assays against Gram-positive, Gram-negative, and fungal strains. Toxicity was evaluated using the HET-CAM assay, while cytocompatibility was determined in fibroblasts, MG-63 cells, and mesenchymal stem cells. Diclofenac-stabilized AgNPs were incorporated into electrospun PCL/PEO nanofibers to generate a functional nanocomposite system. Results: All AgNPs exhibited a characteristic SPR at ~400 nm and high colloidal stability. Diclofenac-stabilized AgNPs (dc-AgNPs) showed the highest antimicrobial activity, with MIC values of 18.8 mg/L against Staphylococcus aureus and Pseudomonas aeruginosa, and 4.7 mg/L against Candida albicans, along with strong bactericidal effects. HET-CAM assays indicated negligible irritation at concentrations up to 75 mg/L. Cytocompatibility results revealed a dose-dependent response, with fibroblasts being more sensitive. Electrospun nanofibers loaded with dc-AgNPs achieved a 2.6 log reduction against Streptococcus mutans and moderate reductions (0.4–0.7 log) against other pathogens. Conclusions: Ligand engineering critically influences the antimicrobial efficacy and biocompatibility of AgNPs. The incorporation of dc-AgNPs into electrospun nanofibers represents a promising approach for treating biofilm-associated wound infections.

Pharmaceutics doi: 10.3390/pharmaceutics18060710

Authors: Lashyn N. Kiyekbayeva Serzhan E. Mombekov Moldir K. Kudaibergenova Nursulu Z. Akhtayeva Assem T. Mamurova Ayala S. Mukhametzhan Yelzhas Nurlykhan Rizvangul B. Ayupova Galiya S. Ibadullayeva Yelena V. Sitdikova Gulnaz N. Musina

Background: Nanophyton iliense U.P. Pratov is a Central Asian halophytic plant whose phytochemical composition and suitability for pharmaceutical formulation remain insufficiently explored. This study evaluated the chemical profile, antioxidant activity, preliminary safety, and gel-forming potential of a hydroethanolic extract of N. iliense for topical application. Methods: The extract was characterized by GC–MS and HPLC. Total polyphenol content was determined, antioxidant activity was assessed using FRAP and DPPH assays, and preliminary cytotoxicity was evaluated using the Artemia salina lethality assay. Results: GC–MS and HPLC analyses showed that the extract contained both lipophilic constituents, including terpenoid and phytosterol-related compounds, and phenolic compounds such as catechin, epicatechin, and naringin. The total polyphenol content reached 485.05 mg GAE/L, exceeding the values obtained for the other plant extracts analyzed under the same conditions. The extract showed concentration-dependent antioxidant activity in both FRAP and DPPH assays. In the DPPH assay, radical scavenging activity increased up to 90.06% at 1.0 mg/mL, while FRAP results confirmed a strong reducing capacity. In the Artemia salina assay, no cytotoxic effect was observed at the tested concentrations. To assess pharmaceutical applicability, five gel formulations were prepared and compared. Gel No. 4, containing N. iliense extract, Lecigel, glycerin, Tween 80, benzyl alcohol, and purified water, showed the most suitable organoleptic and technological characteristics, including homogeneity, good spreadability, and absence of greasy residue. Conclusions: The obtained results indicate that N. iliense extract can be incorporated into a semi-solid formulation, while the extract itself demonstrates relevant in vitro antioxidant properties.

Pharmaceutics doi: 10.3390/pharmaceutics18060709

Authors: Belgin Sever Halilibrahim Ciftci

Background/Objectives: Chronic myeloid leukemia (CML) is primarily associated with the BCR:ABL1 fusion protein. Although tyrosine kinase inhibitors (TKIs) have markedly enhanced treatment outcomes, the development of agents with improved therapeutic characteristics remains necessary. The present work focused on the synthesis of a new series of thiazolone derivatives (F1-11) and the assessment of their anti-CML activity through inhibition of ABL tyrosine kinase (TK). Methods: The designed compounds were prepared through a multistep synthetic pathway involving the formation of a new chalcone intermediate (A), synthesis of a new pyrazoline carbothioamide intermediate (B), and cyclization with different aldehydes to produce the target new thiazolone derivatives (F1-11). Cytotoxic effects were investigated against K562 CML cells using the MTT assay. The lead compound was additionally evaluated in HL-60 AML cells and normal PBMCs. Apoptotic induction was analyzed using Annexin V/ethidium homodimer staining, whereas ABL TK inhibitory activity was measured through the ADP-Glo assay. Molecular docking studies were conducted to explore ligand interactions within the ATP-binding domain of ABL TK. Results: Among the synthesized molecules, F-4 demonstrated the strongest activity against K562 cells with an IC50 value of 6.85 µM, close to that observed for imatinib (IC50 = 5.20 µM). The compound showed reduced cytotoxicity toward HL-60 cells (IC50 = 33.44 µM) and exhibited favorable selectivity toward PBMCs (SI = 13). Apoptosis studies revealed 51% early apoptotic cells and 43% late apoptotic cells following treatment. In the kinase assay, F-4 inhibited ABL TK activity by 39% at 10 µM and by 70% at 100 µM. Docking simulations suggested interactions with residues His361 and Asp381 in addition to nearby hydrophobic amino acids, although the interaction network was less extensive than that of imatinib. Conclusions: The findings identify F-4 as a promising new thiazolone-derived scaffold with selective anti-CML activity and notable ABL TK inhibitory potential. Additional structural optimization may further enhance its binding characteristics and therapeutic efficacy.

Pharmaceutics doi: 10.3390/pharmaceutics18060708

Authors: David O. Oluwole Robert Lees Sneha Banerjee Will Buchanan Lian X. Liu

Background: The formation of wounds or scars often compromises skin anatomy and function, necessitating effective management to restore tissue integrity. Current interventions, including wound debridement, hyperbaric oxygen therapy, antibiotics, wound dressings, and surgical procedures, can be effective but are sometimes limited by high costs and the increasing prevalence of drug resistance. These challenges highlight the need for innovative, cost-effective, and therapeutic alternatives. Method. Our earlier studies assessed the wound closure of wounded human-equivalent epidermal full-thickness skin model (HEFT-SM) with a limited number of Phytoceutical® retinol micellar formulations for a six-day treatment and found that the retinol micellar formulation accelerated wound closure significantly. In this work, three different types of Phytoceutical® retinoid formulations, namely 0.3% retinol, 0.3% retinaldehyde, and 0.03% retinoic acid on the early-stage wound healing efficacy and its collagen structure were studied. Haematoxylin and eosin (H&E) staining analysis was used to assess the wound repair of the 3 mm punch wound after two days and the wound healing efficacy defined as the wound diameter contraction in percentage was assessed. The collagen matrix was examined through the use of Masson’s trichrome staining and confocal laser scanning microscopy (CLSM) for both qualitative and spatial assessment. Results. All formulations promoted wound contraction, with efficacy ranging from 15 ± 1% to 35 ± 2% after two days. The 0.3% retinol micelles showed the highest activity (35 ± 2%), followed by retinaldehyde (32 ± 3%) and retinoic acid (15 ± 1%). In addition, all treatments appeared to stimulate collagen architectural changes suggestive of remodelling activity. Conclusions. The enhanced wound healing observed may be attributed to increased cellular proliferation and migration within the wound microenvironment, supporting epidermal differentiation and tissue stratification. Furthermore, this work showed that combination of Masson’s trichrome staining and confocal laser scanning microscopy (CLSM) is a novel approach for qualitative and spatial assessment of collagen structure.

Pharmaceutics doi: 10.3390/pharmaceutics18060707

Authors: Peng Shao Qiyu Feng Fangfang Pan Juan Zhang Yalan Guan Xiaoxia Sheng Jinqi Zheng

Background: Lidocaine and prilocaine cream is a compounded topical anesthetic formulation comprising lidocaine and prilocaine. Upon application, the active ingredients are locally released and permeate into the subcutaneous tissue, exerting anesthetic effects by blocking ion channels involved in nerve impulse transmission. Variations in drug permeation may influence the onset time, anesthetic efficacy, and duration of action. Methods: This study investigates the in vitro properties of the innovator formulation Emla® and five generic formulations through in vitro bioequivalence studies and Q3 Characterization tests. Results: With the exception of TS, the generic formulations exhibited notable differences compared to the innovator product. Specifically, TB and TL demonstrated significantly higher in vitro release than Emla®, yet exhibited lower in vitro permeation rates. In contrast, TH and TT showed release rates comparable to Emla®, while their permeation rates were similarly reduced. Conclusions: These findings indicate that in vitro release rate does not directly predict in vitro permeation. Permeation behavior is significantly influenced by emulsion globule size, rheological characteristics (viscosity and elasticity), and pH, collectively underscoring the multifactorial nature of this phenomenon.

Pharmaceutics doi: 10.3390/pharmaceutics18060706

Authors: Exequiel O. J. Porta Dana F. AlKharboush Lauren Jackson Felix Pang Aylin Darin Joy Louka Xinyue Shi Geoffrey Wells Frank Kozielski

Although antiviral development against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been dominated by replication-directed strategies, structural and accessory proteins offer a complementary and increasingly important opportunity for small-molecule intervention. These proteins control key processes outside the core replication machinery, including viral entry, membrane remodelling, virion assembly, egress, and host immune modulation, thereby expanding the mechanistic scope of antiviral design. However, many of these targets are membrane-associated, oligomeric, conformationally dynamic, or function through protein–protein interactions, creating distinct challenges in target validation, assay design, and chemical optimisation. In this review, we comprehensively and critically evaluate the structural and accessory proteomes of SARS-CoV-2, with a strict focus on small-molecule tractability and translational relevance. We highlight the most credible direct-acting opportunities, focusing on the membrane (M), envelope (E), and nucleocapsid (N) structural proteins, together with the accessory protein open reading frame 3a (ORF3a), for which emerging chemical matter strengthens confidence in druggability. In contrast, Spike (S) and several host-interface accessory proteins, including ORF6, ORF8, ORF9b, and ORF10, are best viewed as more selective or earlier-stage opportunities that require stronger on-target chemical validation. Emphasis is placed on structural accessibility, mechanism-based assay systems, evidence quality, cellular and in vivo activity, and developability constraints relevant to exposure at the infection site. Rather than replacing replication-directed antivirals, these non-canonical targets are best considered adjunctive or complementary components of future combination strategies designed to broaden antiviral coverage, enhance robustness, and improve pandemic preparedness.

Pharmaceutics doi: 10.3390/pharmaceutics18060705

Authors: Ekaterina Sleptsova Alina Khuzina Daria Sachkova Diana Yuzhakova Yevgeniya Sannova Konstantin Yashin Nina Peskova Svetlana Lermontova Ilya Shchechkin Larisa Klapshina Irina Balalaeva Victoria Turubanova

Background: Photodynamic therapy (PDT) leading to immunogenic cell death (ICD) may serve as a promising basis for the development of antitumor therapeutic strategies. However, the mechanisms of action of photoinduced ICD in primary tumor cultures, including human glioma, remain unexplored. Methods: In the present study, the features of regulated cell death induced by photodynamic therapy using a previously described ICD inducer, porphyrazine III (pz III), were investigated. Cell death was studied in 7 primary cultures of high-grade human gliomas (astrocytomas, oligodendrogliomas, and glioblastomas). Results: Accumulation of porphyrazine III was observed in the endoplasmic reticulum (ER), Golgi apparatus, lysosomes, and mitochondria; however, the distribution of the photosensitizer varied across different cultures. A narrow concentration window of porphyrazine III was established to effectively reach IC85, primarily inducing ferroptosis with contributions from apoptosis and necroptosis accompanied by superoxide anion generation and mitochondrial dysfunction. Conclusions: Given the immunogenic potential of ferroptosis, apoptosis and necroptosis we hypothesize that the induction of PDT using porphyrazine III in glioma will trigger immunogenic cell death.

Pharmaceutics doi: 10.3390/pharmaceutics18060704

Authors: Kuo-Feng Huang Te-Sun Chou Jong-Kai Hsiao

Background/Objectives: Stroke remains the second leading cause of death worldwide, and cell therapy is among the most actively investigated strategies for its treatment. Recent transcriptomic evidence has revealed that 293T cells—the most widely used transient transfection model—possess a neural crest/neuronal lineage, making them a candidate for acute neural tissue engineering. Methods: We implanted iron oxide nanoparticle-labeled 293T cells (293T-ION) into an ischemic rat brain and monitored them longitudinally by 7T MRI, using ION-labeled bone marrow-derived mesenchymal stem cells (rMSC-ION) as a direct comparison. Functional recovery was assessed via mNSS and corner test scores, and infarct size was quantified by MRI. Results: 293T-ION cells showed no migration throughout the 40-day observation period, and functional recovery plateaued early compared with the progressive improvement seen with rMSC-ION. 293T cell implantation provoked pronounced, localized CD68-positive microglial hyperactivation at both implantation and ischemic sites, without migration toward the choroid plexus (CP). In contrast, rMSC-ION actively migrated to the CP and drove superior neuroplasticity marker expression (Ki67, Nestin, NeuN). Conclusions: 293T cells produce transient localized microglial activation and limited brain plasticity, whereas rMSCs drive sustained neurorestoration. Synergistic co-administration of these cell types may represent a future therapeutic strategy bridging hyper-acute and chronic recovery phases.

Pharmaceutics doi: 10.3390/pharmaceutics18060703

Authors: Yura Choi Mi-Young Jung Choul Yong Park

Corneal endothelial dysfunction is a major cause of corneal blindness worldwide. This is primarily due to the limited regenerative capacity of human corneal endothelial cells (CECs) and the global shortage of donor tissues. Corneal endothelial cell therapy (CECT), which involves injecting cultured CECs into the anterior chamber, has emerged as a promising alternative to conventional transplantation. However, its clinical efficacy remains limited by several factors, including rapid cell loss, non-uniform distribution, and insufficient long-term adhesion following injection. Recent advances in biomaterials and regenerative engineering have led to the development of emerging biomaterial-assisted strategies aimed at addressing these challenges. In this review, we provide a mechanistic and translational overview of next-generation CECT, highlighting a range of biomaterial-assisted strategies aimed at improving cell retention, spatial localization, and long-term adhesion following injection. These emerging approaches aim to mitigate key limitations of conventional cell injection therapy, including variability in cell distribution and retention. However, their effectiveness and translational feasibility remain under active investigation. In addition, we analyze recent global patent trends, regulatory frameworks, and market dynamics to highlight emerging opportunities for innovation and development in this field. Although many of these technologies remain at the preclinical or early translational stage, these approaches may provide a promising direction to improve engraftment efficiency, reduce surgical variability, and enable more scalable, minimally invasive treatment options. This review highlights the potential of biomaterial-assisted CECT as a next-generation regenerative strategy and outlines key challenges that must be overcome for successful clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18060702

Authors: Abdelrahman Y. Sherif Mohamed A. Ibrahim

Background/Objectives: Gliclazide’s limited water solubility restricts its absorption across the gastrointestinal tract and compromises its therapeutic performance. This study developed a thermoresponsive solid dispersion based on the inverted thermoresponsive behavior of poloxamer 188 in propylene glycol. Methods: A solubility study was conducted to select components for the thermoresponsive solid dispersion. An I-optimal mixture design was used to optimize the concentrations of the thermoresponsive solid dispersion components (poloxamer 188, propylene glycol, and labrasol). FTIR and XRD were used to investigate the mechanism underlying the inverted thermoresponsive behavior. Finally, the influence of the thermoresponsive solid dispersion on gliclazide dissolution was evaluated through in vitro dissolution testing. Results: Surfactant screening identified labrasol as the optimal surfactant owing to a superior increase in gliclazide solubility compared to propylene glycol alone (2.29-fold). The optimized thermoresponsive solid dispersion (poloxamer 188, propylene glycol, and labrasol at 13.89, 21.43, and 64.68% w/w, respectively) achieved a drug solubility of 10.68 mg/g and a phase transition temperature of 36 °C. XRD and FTIR confirmed that hydrogen bonding is responsible for the system’s conversion between the solid and liquid states. Compared with raw gliclazide, the optimized formulation demonstrated an 8.4-fold increase in the initial dissolution rate and significantly improved dissolution efficiency from 21.77 ± 4.74% to 74.85 ± 2.33%. Conclusions: The present thermoresponsive solid dispersion provides a green alternative to conventional solid dispersion techniques. It avoids reliance on organic solvents, processing that demands high energy input, and additional post-processing operations.

Pharmaceutics doi: 10.3390/pharmaceutics18060701

Authors: Sanjida Ahmed Srishti Paromita Paul Pinky Diponkor Kumar Shill Vidya Surti Jelena M. Janjic

Background: Overexpression of immune cell populations leads to self-amplifying cytokine loops, contributing to chronic inflammation in both allograft rejection and autoimmune conditions. Tacrolimus (TAC), despite being a potent immunosuppressant, has limitations; its systemic adverse effects include nephrotoxicity, neurotoxicity, and high variability in tissue exposure in patients. Currently available therapeutic options are limited by the lack of targeted and localized drug delivery systems, resulting in ineffective control over drug-release behavior. Moreover, TAC being highly lipophilic poses challenges for formulation development. To address these gaps, this study focuses on developing a thermoresponsive hydrogel platform comprising distinct nanocarriers for localized delivery of TAC. The nanocarriers include nanoemulsion (NE) and micelles as TAC carriers, and their particle sizes are specifically engineered at the nanoscale for differential release behavior and to support immune cell targeting (macrophages and T-cells). Incorporation into a thermoresponsive hydrogel matrix enables it to act as a local depot at the injection site and deliver TAC with a slow, extended-release profile. Methods: TAC was loaded into a coconut-rich lipid-phase-based NE via high-pressure microfluidization. Simultaneously, TAC-loaded micelles were optimized using a full-factorial design of experiments (DoE) and manufactured via the thin-film hydration method. Both nanocarriers were evaluated for long-term colloidal stability assessments. Hydrogels were produced maintaining aseptic conditions for sterile batch production. Rheological characterization was performed to assess sol-gel transition, thermoreversibility, and injectability, and in vitro release studies were conducted to evaluate TAC diffusion from the developed nanoformulations. Results: Developed nanocarriers resulted in distinct particle sizes in NE (80–85 nm) and micelles (15–17 nm) with successful TAC loading maintaining long-term colloidal stability. The developed TAC-loaded dual-nanocarrier hydrogel (Dual-HG) showed thermoresponsive behavior and gelation at 37 °C, forming as a local depot. In vitro release studies showed slow and extended tacrolimus release from hydrogels and demonstrated particle size-dependent release behavior between the NE and micelle. Conclusions: Therefore, our study highlights a novel dual nanocarrier hydrogel platform combining TAC-NE and TAC-micelle for localized delivery. The findings support that nanocarriers can be engineered to modulate drug diffusion behavior. Notably, the dual nanocarrier within a thermoresponsive hydrogel platform can be used to deliver one or multiple drugs locally, minimizing systemic exposure when sustained local immunosuppression is required. The 25 mL scale sterile batch production of hydrogels emphasizes their suitability for future translational applications.

Pharmaceutics doi: 10.3390/pharmaceutics18060700

Authors: Yuliia Maslii Nataliia Herbina Olena Ruban Jurga Bernatoniene

Background: Existing therapies for xerostomia are primarily symptomatic, providing temporary mucosal hydration without addressing underlying pathological changes in the oral cavity. In this context, medicated chewing gums containing ascorbic acid and lysozyme hydrochloride offer a promising approach, combining antimicrobial, antioxidant, and trophic effects with physiological salivary stimulation and prolonged local delivery. Methods: For the development of compressed chewing gum formulation, the physicochemical (particle size distribution, moisture absorption capacity, and microscopic characteristics) and technological (flowability, angle of repose, bulk and tapped density, Carr’s index (CI), and Hausner ratio (HR)) properties of the active substances and their formulations with excipients were evaluated. Pharmacological activity was assessed in an atropine-induced xerostomia rat model. Results: The physical mixture of all components showed inferior flow properties compared with the formulation containing pre-granulated lysozyme hydrochloride, as evidenced by higher Carr’s index and Hausner ratio values (CI = 17, HR = 1.20 vs. CI = 13, HR = 1.14), indicating improved processability after pre-granulation. The effect of relative humidity during formulation was also assessed, with an optimal level of 40% required to ensure process stability due to the hygroscopic nature of the components. Based on these data, technological approaches ensuring processability were established, including wet pre-granulation of lysozyme hydrochloride and premixing of ascorbic acid to reduce oxidation risk. These approaches resulted in an optimized compression mass with excellent flowability (CI = 8, HR = 1.09), suitable for the preparation of medicated chewing gum. An optimal compression force (7 kN) ensured suitable rheological and textural properties, resulting in rapid and nearly complete release of the active ingredients from the medicated chewing gum, consistent with kinetic analysis. In vivo studies using an atropine-induced xerostomia rat model demonstrated that the combination of ascorbic acid and lysozyme hydrochloride significantly increased salivary secretion (2.17-fold vs. control pathology group) and reduced salivary gland mass coefficients (by 13–18% compared with the control pathology group and groups receiving individual active ingredients), alongside improvement of oxidative stress markers, including a reduction in TBA-reactants (by 51.6%) and an increase in catalase activity (by 51.0%). Conclusions: The developed medicated chewing gum showed favorable technological properties, efficient release of active ingredients, and anti-xerostomic activity in vivo, indicating its potential for xerostomia relief and oral health support.

Pharmaceutics doi: 10.3390/pharmaceutics18060699

Authors: Hiroko Otake Rie Tanaka Fumihiko Ogata Yosuke Nakazawa Manju Misra Kazutaka Kanai Masanobu Tsubaki Naoki Yamamoto Naohito Kawasaki Noriaki Nagai

Background/Objectives: Dry eye disease (DED) is a multifactorial ocular surface disorder characterized by tear film instability and decreased tear secretion, largely driven by chronic ocular surface inflammation. Although current therapies primarily target inflammation and tear film stabilization, their clinical efficacy is often limited by insufficient ocular surface retention. In this study, we explored a drug repositioning strategy for DED by developing a nanocrystalline formulation of troxipide (TRO), a gastric mucosal protective agent with cytoprotective properties. Methods and Results: A TRO nanosuspension (TRO-NPs) was successfully prepared by wet bead milling, yielding particles with a mean diameter of approximately 100 nm. Physicochemical characterization revealed that the crystalline structure, solubility, viscosity, pH, and osmolarity of the nanosuspension were comparable with those of the conventional TRO microsuspension (TRO-MPs). In contrast, the TRO-NPs exhibited markedly improved dispersion stability, maintaining particle suspension for at least 1 month after preparation. Repeated topical instillation of the TRO-NPs did not induce corneal toxicity or inflammation in rabbits, and resulted in significantly higher drug retention in the tear fluid than that observed for the TRO-MPs. Furthermore, in an N-acetylcysteine-induced rabbit dry eye model, repetitive instillation of the TRO-NPs significantly increased tear volume and mucin levels, leading to improved tear film stability. Conclusions: These findings demonstrate that nanosuspension-based formulations can enhance ocular surface retention and therapeutic efficacy of TRO. TRO-NPs therefore represent a promising nanomedicine-based repositioned therapy for the treatment of DED.

Pharmaceutics doi: 10.3390/pharmaceutics18060698

Authors: Katarina Sokač Pogrmilović Gordana Matijašić Krunoslav Žižek

Achieving precise control over anticancer drug release remains one of the key challenges in modern pharmaceutical development, as it directly determines therapeutic efficacy, systemic toxicity, and patient outcomes. This review critically evaluates recent advances in three major formulation strategies: polymeric solid dispersions, cyclodextrin-based inclusion complexes, and metal–organic frameworks (MOFs), with a particular focus on their capacity to tailor anticancer drug release. Over the past decade, polymeric solid dispersions and cyclodextrin-based carriers have played a central role in improving the dissolution and bioavailability of poorly water-soluble anticancer agents, while also enabling modified release profiles through rational formulation design. Increasing structural complexity, including ternary systems and supramolecular assemblies, reflects a shift toward more controllable delivery platforms. In recent years, MOFs have emerged as highly adaptable porous materials capable of supporting controlled and stimuli-responsive release. The integration of imaging agents, magnetic components, and photothermal functionalities has further enabled the design of multifunctional and theranostic platforms. Taken together, these technologies reflect a shift from conventional solubility enhancement toward structurally engineered systems designed to achieve predictable and controlled drug release. Continued advances in material design and formulation strategies are expected to further refine release kinetics and support the development of next-generation anticancer therapies aligned with the growing demand for precision medicine.

Pharmaceutics doi: 10.3390/pharmaceutics18060697

Authors: Junha Lee Suan Kwon Yoosoo Yang Jiwoong Choi

Extracellular vesicles (EVs) have attracted considerable attention as natural nanocarriers for immune modulation owing to their intrinsic biocompatibility, nanoscale size, and capacity to transport diverse bioactive cargos. In inflammatory diseases, EV-based therapeutics provide unique opportunities to regulate dysregulated immune responses; however, their clinical translation remains constrained by limited cell-specific targeting efficiency and uncontrolled biodistribution. Achieving precise and selective delivery to immune cells and other inflammation-associated cellular components within diseased tissues is therefore critical for maximizing therapeutic efficacy while minimizing off-target effects. This review comprehensively summarizes recent advances in cell-specific EV-targeting strategies for immune modulation in inflammatory diseases, with a particular focus on active targeting approaches enabled by EV surface engineering. A range of targeting ligands, including antibodies, peptides, aptamers, glycans, and membrane proteins, is discussed in the context of enhancing selective interactions between EVs and specific immune cell subsets. Special emphasis is placed on cell-directed targeting strategies toward diverse immune cell populations, including macrophages and T cells, highlighting how rational control of EV–cell interactions can be utilized to reprogram immune phenotypes, suppress pathological inflammation, and restore immune homeostasis. Accordingly, this review integrates recent progress in cell-specific EV targeting into a coherent conceptual framework, which may assist researchers in the rational design of EV-based immunomodulatory therapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18060696

Authors: Bruna Rojas Fróes Juliana Guanaes Pina Mariana da Mata Alves Alquiandra S. F. Mançano Fernanda C. Cardoso Juliana Mozer Sciani

Background/Objectives: Alzheimer’s disease (AD) is characterized by beta-amyloid (Aβ) plaque deposition, which impairs several cellular processes, including autophagy. Considering the multifactorial nature of AD, the development of therapies acting on alternative molecular targets is necessary. In this study, we evaluated the neuroprotective effect of a molecule from the hydrozoan Eudendrium carneum and investigated its impact on autophagy-related pathways. Methods: The secretion of E. carneum was fractionated by RP-HPLC according to its neuroprotective activity in SH-SY5Y cells exposed to oAβ42, evaluated using LDH and MTT assays. The purified molecule (named EC5), characterized by mass spectrometry, was evaluated regarding in silico toxicity and calcium dynamics. Neuronal lysosomal morphology was assessed using the LysoTracker probe, and cathepsin D activity was determined using a synthetic substrate. The expression of autophagy-related proteins (mTOR, LAMP-1, and LC3B) was evaluated by dot blotting, and amyloid plaque clearance was quantified using Thioflavin-T staining. Results: The steroid glycoside putatively identified as Sarmentoside B (EC5) exhibited neuroprotective effects and showed no toxicity or alterations in neuronal calcium or sodium channel dynamics. EC5 restored lysosomal morphology and cathepsin D activity, reversing the impairment induced by oAβ42. Furthermore, EC5 reduced mTOR expression, and this interaction was supported by molecular docking analysis. Lysosomal restoration promoted the clearance of oAβ42 aggregates, as evidenced by Thioflavin-T staining, resulting in reduced neuronal death. Conclusions: EC5, putatively identified as Sarmentoside B, exerts neuroprotective effects against oAβ42-induced toxicity by promoting autophagy-related amyloid clearance, highlighting its therapeutic potential for AD.

Pharmaceutics doi: 10.3390/pharmaceutics18060695

Authors: Boqin Ma Yan Li Jiahui Zhang Yuanlei Fu Haiqiang Cao

Objective: This study aimed to develop an oral celastrol-loaded self-microemulsifying drug delivery system (Cel-SMEDDS) to enhance the therapeutic efficacy against rheumatoid arthritis and reduce toxicity. Methods: The optimal Cel-SMEDDS formulation, identified through solubility screening, excipient compatibility assays, and pseudo-ternary phase diagram analysis, was characterized by particle size, PDI, zeta potential, in vitro release, and stability. In vitro anti-inflammatory activity was evaluated in LPS-induced RAW264.7 macrophages, while in vivo anti-RA efficacy was assessed in CIA mice via paw swelling, clinical scoring, serum TNF-α, and joint histopathology. Preliminary safety was examined by hematological, serum biochemical, and histopathological analyses in mice. Results: The optimal Cel-SMEDDS formulation consisted of LABRAFIL M 1944 CS-Kolliphor RH40-CAPRYOL 90 (0.2:0.48:0.32, w/w/w) with a drug loading of 1.5% (w/w). It spontaneously formed uniform microemulsions with a mean particle size of 26.70 nm, PDI of 0.067, and zeta potential of −2.87 mV. In vitro, Cel-SMEDDS showed enhanced cytotoxicity against M1-type macrophages (IC50 = 0.1753 μg/mL vs. 0.2684 μg/mL for free Cel), significantly suppressed pro-inflammatory TNF-α and IL-1β expression, and upregulated anti-inflammatory IL-10. In CIA mice, oral Cel-SMEDDS reduced paw swelling by 37.42% (vs. 22.79% for free Cel), markedly decreased serum and intra-articular TNF-α levels, and alleviated articular cartilage damage. Preliminary safety evaluation demonstrated no significant abnormalities in hematological parameters, liver/kidney function, or major organ histology. Conclusions: The optimized oral Cel-SMEDDS effectively inhibits the expression of pro-inflammatory cytokine TNF-α both in vitro and in vivo, exhibits superior anti-RA activity compared to free Cel, and possesses favorable safety. This formulation addresses the key limitations of celastrol and shows promising potential for clinical translation in RA treatment.

Pharmaceutics doi: 10.3390/pharmaceutics18060694

Authors: José Márcio Fernandes da Silva Dominique Mesquita e Silva Danilo de Souza Costa Monique C. Amaro Rayssa A. Cajas Josué de Moraes Guilherme Diniz Tavares Ademar Alves Da Silva Filho

Background: Schistosomiasis remains a major neglected tropical disease, with praziquantel (PZQ) as the only widely used treatment, despite its limitations. Parthenolide (PTL), a sesquiterpene lactone, exhibits potent in vitro antischistosomal activity; however, its poor aqueous solubility, low oral bioavailability, and chemical instability may limit its in vivo efficacy. Objective: This study investigated whether nanoencapsulation in nanostructured lipid carriers (NLC) could enable the in vivo antischistosomal activity of PTL. Methods: PTL was isolated from Tanacetum parthenium and incorporated into NLC using hot emulsification followed by ultrasonication. The resulting formulation (NLC-PTL) was physicochemically characterized, and its in vivo antischistosomal efficacy was evaluated in a murine model of Schistosoma mansoni infection. Results: NLC-PTL exhibited nanoscale size, low polydispersity, high encapsulation efficiency, and sustained drug release. In vivo, free PTL showed no significant effect on worm burden, whereas NLC-PTL achieved a marked reduction (77.9%) in adult worms and significantly decreased egg output compared to controls (p < 0.001). Blank NLC had no antiparasitic effect. Conclusions: Nanoencapsulation was associated with in vivo antischistosomal activity of PTL compared to the free compound. These findings suggest that formulation strategies may influence the in vivo performance of lipophilic natural products in schistosomiasis.

Pharmaceutics doi: 10.3390/pharmaceutics18060693

Authors: Exequiel O. J. Porta Dana F. AlKharboush Lauren Jackson Felix Pang Aylin Darin Joy Louka Mohammed Quamruzzaman Xinyue Shi Geoffrey Wells Frank Kozielski

The SARS-CoV-2 non-structural proteome remains the most clinically validated and strategically important landscape for direct-acting small-molecule antiviral drug discovery. The success of inhibitors targeting the main protease (Mpro, Nsp5) and RNA-dependent RNA polymerase (RdRp, Nsp12) has firmly established viral replication enzymes as tractable, druggable, and therapeutically relevant targets, while setting clear benchmarks for translational antiviral development. Building on this foundation, a second wave of non-structural protein (Nsp) targets has emerged with increasing translational promise, including the papain-like protease (PLpro), the bifunctional Nsp14 proofreading and capping machinery, Nsp16 2′-O-methyltransferase, Nsp13 helicase, and Nsp15 endoribonuclease. In parallel, additional components such as Nsp1 and the Mac1 domain of Nsp3 continue to expand the antiviral design space, although they remain at earlier stages of chemical validation. In this review, we comprehensively assess SARS-CoV-2 non-structural proteins through a medicinal chemistry and translational lens, with an emphasis on structural tractability, mechanism of action, quality of chemical matter, cellular and in vivo antiviral evidence, evolutionary conservation, resistance liabilities, and developability. Particular attention is given to the features that distinguish tool compounds from genuinely actionable leads and to the opportunities for rational combination regimens that extend beyond first-generation protease- and polymerase-centred therapy. Collectively, the non-structural proteome offers the strongest foundation for next-generation and potentially broader-spectrum coronavirus antivirals with improved resilience to viral evolution.

Pharmaceutics doi: 10.3390/pharmaceutics18060692

Authors: Chao-Yi Lu Lara Vaid Asha Adler Gulcin Arslan Azizoglu Andrey V. Romanyuk Mark R. Prausnitz

Background/Objectives: Drug administration by microneedle patch (MNP) offers advantages over conventional dosage forms as a painless, self-administered skin patch for parenteral delivery. Dissolvable MNPs are typically manufactured by casting an aqueous formulation containing dissolved active pharmaceutical ingredient (API) and excipients into a mold and allowing it to dry. This process can be detrimental to APIs that are sensitive to dissolution and drying during the casting process. Methods: This study presents a MNP fabrication process in which drug particles are suspended in an organic solvent carrier without being dissolved in the solvent. Results: We started with drug particles either as pure API or formulated with excipients to stabilize them. We then screened nine organic solvents, ranging from high (methanol) to low (toluene) polarity, to identify those that suspend the drug particles without dissolution or damage to the API. To guide formulation of stabilized drug particles, we generated a companion database of 16 common stabilizing excipients and measured their solubility in our panel of organic solvents to identify excipient–solvent combinations that did not lead to excipient dissolution. We generated a second database of 14 water-soluble polymers to serve as the microneedle matrix material and determined their solubility in our panel of solvents to identify solvents that enabled polymer dissolution. Using these data, we designed casting solutions that suspended particles of API (and excipients) in an organic solvent that dissolved a matrix polymer. Casting and drying these solutions on molds produced MNPs for delivery of three model compounds: lyophilized tetanus toxoid (i.e., a vaccine), methotrexate (i.e., a small molecule drug), and insulin (i.e., a biologic). Conclusions: We conclude that this fabrication method, guided by the excipient and polymer solubility databases, offers a novel method to produce MNPs by suspension casting of drugs in organic solvents.

Pharmaceutics doi: 10.3390/pharmaceutics18060691

Authors: Razvan Ghiarasim Alexandru Rotaru Cristian-Dragos Varganici Mariana Pinteala Narcisa-Laura Marangoci Ion Tiginyanu Natalia Simionescu

Background/Objectives: Superparamagnetic iron-oxide nanoparticles (SPIONs) bearing poly(methacrylate) brushes were synthesized via surface-initiated atom-transfer radical polymerization (SI-ATRP) as magnetically responsive nanoplatforms. Three brush architectures, poly(2-hydroxyethyl methacrylate) (PHEMA) and poly(poly(ethylene glycol) methacrylate) with six ethylene-oxide units (PPEGMA6) and ten units (PPEGMA10), were grown from a dopamine-anchored initiator and covalently loaded with methotrexate (MTX). Methods: Physicochemical characterization confirmed successful polymer grafting, tunable hydrodynamic size (185–1320 nm before MTX conjugation and 427–694 nm after), retained superparamagnetic properties (22–69 emu g−1), and high drug payloads, with PPEGMA6 achieving 131 µg mg−1. MTX conjugation induced partial compaction of the polymer shell yet maintained ζ-potentials conducive to colloidal stability. Results: In vitro assays showed negligible toxicity toward primary human fibroblasts, whereas MTX-decorated formulations induced a pronounced concentration-dependent cytotoxic effect in MCF-7 breast cancer cells, reaching 69% loss of viability—significantly higher than free MTX. Structure–activity analysis attributes the superior performance of PPEGMA6-MTX to its balanced brush density, high payload, and favorable surface charge. Conclusions: These findings demonstrate that precise modulation of polymer brush architecture via SI-ATRP yields SPION-based nanocarriers that integrate MRI visibility and the potential for magnetic guidance and targeted chemotherapy. The PPEGMA6-MTX construct is highlighted as a promising platform for future preclinical investigations.

Pharmaceutics doi: 10.3390/pharmaceutics18060690

Authors: Imani A. Kirven Patrice Penfornis Muhammad R. Siddiqui Kenneth R. Butler Richard J. Roman Clayton T. Larsen Candace M. Howard Pier Paolo Claudio

Background/Objectives: Immune checkpoint inhibitors targeting the PD-1/PD-L1 axis have transformed cancer treatment, yet therapeutic responses remain limited in many solid tumors due to poor and uneven drug distribution within the tumor microenvironment (TME). Here, we evaluated whether co-formulation of an anti-PD-1 antibody (RMP1-14, murine surrogate for pembrolizumab) with Imagent® microbubble/liposome (MBLP) complexes and ultrasound activation could enhance tumor-specific delivery while reducing systemic exposure. Methods: Immunocompetent MC-38 colorectal tumor-bearing mice (B6(Cg)-Tyrc-2J/J, 7-week-old females) received isotype control, isotype/MBLP/US, RMP1-14 alone, RMP1-14/MBLP, or RMP1-14/MBLP/US. Survival was analyzed by Kaplan–Meier curves, tumor necrosis by H&E staining, antibody biodistribution by immunohistochemistry, and tumor perfusion by laser speckle imaging. Results: No significant differences in tumor size or body weight were observed between groups. Survival analysis showed significant improvements in the RMP1-14 (p = 0.013) and RMP1-14/MBLP/US (p = 0.047) groups versus isotype controls, with the RMP1-14/MBLP/US group achieving the longest mean survival (57.8 days vs. 26.5 days for RMP1-14 alone) and complete tumor regression in 2/8 mice. The RMP1-14/MBLP/US group demonstrated significantly greater tumor necrosis than all other groups. Immunohistochemical analysis confirmed a 6.1-fold increase in intratumoral antibody accumulation with MBLP/US versus RMP1-14 alone (p = 0.0003), alongside significantly reduced off-target exposure in spleen, liver, kidney, and heart. Laser speckle imaging revealed a transient ~30% increase in tumor perfusion during MBLP/US treatment, consistent with cavitation-mediated hemodynamic effects. Conclusions: These findings demonstrate that MBLP/US co-formulation enhances intratumoral delivery of checkpoint inhibitors, improves survival, and reduces systemic organ exposure, representing a promising platform to improve the efficacy and safety profile of antibody-based immunotherapy.

Pharmaceutics doi: 10.3390/pharmaceutics18060689

Authors: Ali Al-Samydai Ali Olamat Arwa Al Khatib Jamal Al Nabulsi Hamdi Al Nsairat Walhan Alshaer Sara Al Mahamid Alaa Alsanabrah Ahmed S. A. Ali Agha Hamza AbuOwida

Objectives: Current liposomal drug delivery studies remain largely formulation-specific and descriptive, with limited predictive capability. This study aimed to develop co-loaded nanoliposomes and establish an integrated framework for predictive analysis of drug release. Methods: PEGylated nanoliposomes co-loaded with bendamustine and rutin were prepared using the thin-film hydration method. Physicochemical properties, encapsulation efficiency, and in vitro release were evaluated. An integrated analytical approach combining data augmentation, monotonicity-constrained denoising, Weibull kinetic modeling, and machine learning was applied to characterize and predict release behavior. Results: Co-loaded formulations exhibited higher encapsulation efficiency (up to 77.75%) and distinct release profiles compared to single-drug systems. Weibull modeling adequately described nonlinear release kinetics (R2 ≈ 0.90–0.94). Machine learning enabled within-formulation prediction of later-stage release from early time points (R2 > 0.98; MAE ≈ 0.83–1.00%), although leave-one-formulation-out cross-validation confirmed that cross-formulation generalization remains limited. Reconstructed release curves captured overall formulation-dependent trends, despite variable accuracy in individual kinetic parameters. Conclusions: The proposed hybrid framework enables early prediction of drug release and reveals that curve-level behavior may be approximated without precise parameter estimation, though this reflects parameter compensability rather than robust prediction. This work provides a proof-of-concept framework for analyzing nanoliposomal drug delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18060688

Authors: Maja Đanić Natalija Dedić Dragana Zaklan Slavica Lazarević Bojan Stanimirov Momir Mikov Nebojša Pavlović

Background: Caffeine, although widely used in dermatological and cosmetic products, exhibits limited permeability through the stratum corneum, highlighting the need for strategies for optimizing delivery. The aim of this study was in vitro investigation of the effects of probiotic bacterial lysates and submicellar concentrations of bile acids on caffeine permeation, with a particular focus on permeation kinetics. Methods: Caffeine permeability was evaluated using the Skin Parallel Artificial Membrane Permeability Assay (Skin-PAMPA). Donor and acceptor concentrations were quantified by HPLC at predefined time points (1, 2, 4, 6, and 12 h), followed by calculation of apparent permeability coefficients, cumulative permeation profiles, and interval permeation rates in systems containing probiotic lysates and submicellar concentrations of cholic acid (CA) or deoxycholic acid (DCA). Results: Probiotic lysates significantly reduced caffeine permeability (0.98 ± 0.02 × 10−6 vs. 1.57 ± 0.14 × 10−6 cm/s in the control group) and modified transport kinetics resulting in lower early-phase interval permeation rates and reduced cumulative permeation. Conversely, bile acids increased the apparent permeability of caffeine, with the highest value observed in the DCA group (2.30 ± 0.08 × 10−6 cm/s). Conclusions: Overall, probiotic lysates and bile acids modulated caffeine permeation across the Skin-PAMPA membrane primarily by reshaping permeation kinetics rather than simply changing overall permeability. Their combined effects may provide a basis for designing topical formulations with tailored permeation profiles.

Pharmaceutics doi: 10.3390/pharmaceutics18060687

Authors: Dipa K. Israni Mansi Shah Heena Chauhan Mumuxa Rathod Bhupendra G. Prajapati Supachoke Mangmool Sudarshan Singh Chuda Chittasupho

Inflammation plays a crucial role in defending the body against harmful stimuli and maintaining physiological balance; however, when it becomes chronic, it contributes to the pathogenesis of several long-term diseases, including autoimmune conditions, cardiovascular and neurodegenerative disorders, and various cancers. Although conventional anti-inflammatory drugs provide symptomatic relief, their long-term use is often associated with adverse side effects. This limitation has shifted scientific attention toward naturally occurring bioactive molecules with potent, safer anti-inflammatory activity. Dietary incorporation of phytopharmaceuticals, such as flavonoids, polyphenols, alkaloids, terpenoids, and fatty acids, has been shown to regulate immune and oxidative mechanisms and to modulate key inflammatory signaling cascades, including the NF-κB, mitogen-activated protein kinase (MAPK), and JAK/STAT pathways. These agents also influence cytokine secretion, NLRP3 inflammasome activation, and antioxidant defense mechanisms involving the Nrf2/HO-1 axis. The current review emphasizes the relevance of major natural plant products in therapy, like quercetin and rutin, resveratrol, glycyrrhizin, lycopene, and indole-3-carbinol. Moreover, recent progress in anti-inflammatory research has focused on novel resolution-based strategies that extend beyond inflammation and oxidative stress suppression. In addition, the review discusses innovations including nanoformulation-assisted targeted delivery, specialized pro-resolving lipid mediators such as resolvins and protectins, and microbiota-oriented therapeutic approaches. Additionally, the review highlights the integration of personalized medicine supported by multi-omics technologies to enhance treatment precision and clinical outcomes. By synthesizing findings from preclinical studies and clinical investigations, this work emphasizes the synergistic therapeutic potential of bioactive compounds from natural sources and resolution-enhancing techniques in restoring immune homeostasis and effectively mitigating chronic inflammation.

Pharmaceutics doi: 10.3390/pharmaceutics18060686

Authors: Miranda Piccioni Giuseppe Curcio Alessandro Graziani Donatella Pietrella

Background: Biofilms consist of complex microbial communities embedded in an extracellular matrix which confer resistance to the most used antimicrobial agents. Chronic wounds are often associated with burns, trauma, surgery, diabetes and peripheral vascular disease. They are characterized by a marked delay in wound healing favoring the development of microbial biofilms, which in turn further delay tissue regeneration. Staphylococcus aureus, Staphylococcus epidermidis, and methicillin-resistant staphylococci biofilms are found in chronic wounds, seriously hindering wound treatment. Vitreoscilla filiformis, a Gram-negative non-pathogenic filamentous bacterium, has been shown to improve atopic dermatitis by reducing S. aureus colonization and inducing antioxidant responses in the skin. Objectives: The aim of the present study was to evaluate the antimicrobial, anti-inflammatory, and regenerative activities of the V. filiformis supernatant (VFS). Methods: The effect of VFS on bacteria growth was assessed by microbial growth kinetics and biofilm formation and dispersal. Antioxidant potential was determined by DPPH-scavenging ability and reduction in intracellular reactive oxygen species (ROS). The regenerative properties were assessed by scratch assay. Results: V. filiformis VFS holds strong anti-biofilm activity against S. aureus, S. epidermidis and methicillin-resistant S. aureus (MRSA), acting during both biofilm formation and dispersion. The decrease in biofilm mass is accompanied by a significant increase in the planktonic form compared to the untreated cells. Moreover, VFS is characterized by an interesting antioxidant activity, as demonstrated by a cell-free DPPH assay and a neutrophil-based in vitro assay. In addition, VFS can stimulate tissue regeneration in human dermal fibroblasts and keratinocytes. Conclusions: The demonstration of anti-biofilm, antioxidant and regenerative properties of V. filiformis supernatant could be exploited for the treatment of biofilm-associated wound infections.

Pharmaceutics doi: 10.3390/pharmaceutics18060685

Authors: Wanzhen Li Chenyu Liao Yuzhen Li Zijun Lin Danni Xiao Gengsheng Ye Yanjuan Huang Chunshun Zhao Shengmiao Cui

Background: Intrauterine adhesions, a leading cause of female infertility, frequently recur in 30–62.5% of patients despite hysteroscopic adhesiolysis and adjuvant therapies. Current intrauterine barriers, including injectable hydrogels, often lack sufficient bioactivity and tissue retention, failing to address the underlying pathological inflammation and oxidative stress driving abnormal fibrosis. Methods: Herein, we tailored a reactive oxygen species (ROS)-responsive, mussel-inspired adhesive injectable hydrogel (OHA-CP@TA) to intelligently modulate the inflammatory niche and promote normal endometrial regeneration. OHA-CP@TA was fabricated through Schiff base bonds between oxidized hyaluronic acid (OHA) and phenylboronic acid-modified carboxymethyl chitosan (CMCS-PBA), and boronate ester bonds between CMCS-PBA and tannic acid (TA). Results: OHA-CP@TA exhibited good mechanical strength, injectability, self-healing, and shear-thinning properties, and importantly, robust and stable adhesion to uterine tissue, overcoming endometrial mucus clearance. It also showed favorable in vivo uterine cavity retention for at least 7 days that covered the critical endometrial repair period. Within the postoperative inflammatory milieu, OHA-CP@TA intelligently released TA in a ROS-dependent manner, which effectively scavenged various ROS and significantly alleviated inflammation, and promoted M1 macrophage polarization into M2 phenotype. This targeted ROS scavenging and immunoregulation inhibited endometrium fibrosis progression, evidenced by downregulation of α-SMA and Col-1, and actively promoted endometrial repair and regeneration, demonstrated by enhanced angiogenesis, increased endometrial thickness, and restoration of glandular numbers. Furthermore, OHA-CP@TA exhibited good biocompatibility, in vivo biodegradability and safety. Conclusions: Therefore, OHA-CP@TA represents a promising, clinically translatable strategy for overcoming the limitations of current IUA management.

Pharmaceutics doi: 10.3390/pharmaceutics18060684

Authors: Davide D’Angelo Stefania Glieca Lisa Flammini Simona Bertoni Annalisa Bianchera Eride Quarta Ben Forbes Fabio Sonvico Francesca Buttini

Background: Cyclosporine is widely used to prevent transplant rejection; however, its systemic administration is associated with low bioavailability and a risk of severe adverse side effects. In the context of lung transplantation, local pulmonary delivery represents a promising strategy to reduce the required dose while enhancing local anti-inflammatory efficacy and limiting systemic toxicity. Methods: In this study, cyclosporine was encapsulated in liposomes coated with calcium phosphate to improve cellular uptake. The liposomal formulation was subsequently converted into a dry powder for inhalation to enable pulmonary administration, combining cyclosporine-loaded liposomes with a calcium phosphate coating, extending prior work on inhaled liposomal cyclosporine and mineral-coated liposomes into a single platform. The cyclosporine loading was optimised to achieve an efficient drug content in the final formulation. Results: The presence of the calcium phosphate coating on the liposomal surface was confirmed by the shift in zeta potential and by cryo-transmission electron microscopy. The resulting dry powder exhibited suitable aerodynamic properties for pulmonary delivery with a fine particle fraction of 33.6 ± 1.6%. In vitro biocompatibility studies performed on A549 epithelial cells and THP-1 monocytic cells demonstrated that the formulation did not affect cell viability. Furthermore, the formulation containing calcium phosphate-coated liposomes showed a stronger anti-inflammatory effect compared with both uncoated liposomal formulations and the corresponding raw material, consisting of a physical mixture of phospholipids and cyclosporine. Conclusions: Overall, despite limitations on respirability and efficacy that will require further in vivo studies, this calcium phosphate-coated liposomal dry powder could represent a promising strategy for targeted pulmonary delivery of cyclosporine, with potential to improve the prevention of lung transplant rejection while minimising systemic side effects.

Pharmaceutics doi: 10.3390/pharmaceutics18060683

Authors: Emily Nghiem Ariel Tzamarot Terence Li Zimo Huang Mahshid Mohammadi Dior Dedushi Yvonne Saenger Fernand Bteich Chaoyuan Kuang

Background/Objectives: PI3K/AKT/mTOR is a key pathway in cell proliferation, metabolism, and survival. Activating PIK3CA mutations are seen in up to 20% of colorectal cancers and are associated with increased cyclo-oxygenase-2 (COX-2) expression. Recent studies demonstrated a significant survival benefit from taking low-dose aspirin, a nonselective COX inhibitor, supporting further exploration of the synergistic effects of combined PI3Kα inhibitor (inavolisib) and COX-2 inhibitor (celecoxib) therapy. Methods: The effects of celecoxib–inavolisib combination treatment were tested on human colorectal cancer cell lines and patient-derived organoid models. Experiments included cell viability and colony formation assays, immunoblotting, and immunofluorescence. Results: We found that celecoxib and inavolisib demonstrated synergy in suppressing the growth of colorectal cancer cell lines, grown in both 2D and 3D cell culture, regardless of PIK3CA mutation status. In patient-derived organoid models, while synergy was seen in both organoids, growth of the PIK3CA mutated organoid was more potently suppressed. Immunoblotting of cells after combination treatment showed decreased expression of mitogenic signaling marker p-AKT across all 2D cell lines and in both cell lines grown as 3D spheroids, as well as increased expression of apoptotic marker cPARP in four out of five 2D cell lines and in both cell lines grown as 3D spheroids. Immunofluorescence staining of organoids after combination treatment, however, showed no significant increase in expression of apoptotic marker Cas-3 nor in mitogenic marker Ki-67 in either organoid. Furthermore, an apoptosis assay performed on two cell lines showed no significant increase in Annexin V or phosphatidylserine staining. Conclusions: Celecoxib and inavolisib demonstrated synergy in suppressing the growth of both colorectal cancer cell lines and patient-derived organoids, though PIK3CA mutation status did not appear to affect drug efficacy in cell lines as it did in patient-derived organoids. Potential compensatory or resistance mechanisms might include oncogene drivers in the MAPK/ERK pathway. When compared to monotherapy, combination therapy was the only drug condition to significantly increase the percentage of apoptotic cells based on Annexin V and phosphatidylserine staining, and this effect was only seen in the PIK3CA mutated cell line. Ultimately, our findings provide preliminary support for celecoxib–inavolisib combination treatment as a rational therapeutic avenue warranting further preclinical investigation.

Pharmaceutics doi: 10.3390/pharmaceutics18060682

Authors: Shiyuan Lin Feixian Lu Qiao Li Kefeng Zhang Wei Zhang Hui Chen Jianxin Wang

Background: Hepatic fibrosis is characterized by the abnormal activation of hepatic stellate cells (HSCs) and excessive deposition of the extracellular matrix. Currently, effective clinical therapeutic strategies remain limited. Modulating ferroptosis-related pathways in activated HSCs has emerged as a promising therapeutic target for hepatic fibrosis treatment. Methods: An amphiphilic copolymer was synthesized by conjugating COS with ART, which spontaneously self-assembled into micelles; subsequent modification with retinoic acid (RA) yielded RA-functionalized ART–COS copolymer micelles. Curcumin was selected as a model drug to evaluate the potential of the micelles in enhancing intestinal epithelial transport, oral absorption and bioavailability. Meanwhile, in vitro targeting ability, capacity to modulate ferroptosis in HSCs and in vivo therapeutic efficacy were systematically investigated. Results: The RA-functionalized ART–COS micelles significantly enhanced intestinal epithelial drug transport, oral absorption, and bioavailability. In vitro experiments demonstrated that the micelles preferentially accumulate in activated HSCs, inhibit GPX4 expression, and induce excessive ROS production and ferroptosis, thereby effectively attenuating hepatic fibrosis. In vivo studies confirmed that the micelles regulated extracellular matrix metabolism, reduced collagen deposition, suppressed the activation and proliferation of HSCs, and ultimately helped attenuate hepatic fibrosis progression. Conclusions: This study successfully developed RA-functionalized ART–COS copolymer micelles. The micelles improve the accumulation of artesunate in liver tissue and yield favorable anti-fibrotic effects, thereby providing a promising translational strategy for anti-fibrotic therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18060681

Authors: Anjani Saxena Yashpal Singh Mumtesh Kumar Saxena Sachin Kumar Meena Mrigesh Aman Kamboj Manish Kumar Verma Manjul Kandpal Satya PalSingh

Background: Salmonella Typhimurium is a major pathogen causing non-typhoidal salmonellosis in humans. Poultry is a major reservoir of S. Typhimurium. Currently available vaccines against S. Typhimurium are not very effective. Therefore, the search for novel adjuvants to improve vaccine efficacy is a priority for developing effective and efficient vaccines. Method: In this study, next-generation adjuvants, such as calcium phosphate nanoparticles, are being evaluated. Our objective was to assess the potential of calcium phosphate nanoparticles, using outer membrane proteins of Salmonella Typhimurium as antigens, for immune-potential testing in poultry, with Montanide as a control. The toxicity of the prepared vaccine formulation was evaluated in rats. Results: CaP-Omp-Nps in the 30–45 nm size range showed a protein entrapment efficiency of 42.5% and a loading capacity of 50.3%. Both vaccinated groups, calcium phosphate outer membrane protein nanoparticles (CaP-Omp-Nps) and Montanide, induced an efficient humoral immune response, with mean titers of 3.48 + 0.0245 and 4.9 + 0.0142 on the 15th day, 3.5 + 0.0118 and 4.79 + 0.009 on the 30th day, and 4.48 + 0.427 and 5.31 + 0.154 on the 45th day post vaccination, respectively, indicating an improvement (CaP-Omp-Nps group) or stability (Montanide group) over the study period. Further, the CaP-Omp-Nps group revealed a better cell-mediated immune response than the Montanide-Omp group. The toxicity study in rats showed no significant differences in serum biomarkers and blood chemistry parameters, indicating that the nano-vaccine formulation is non-toxic and safe. Outer membrane proteins of Salmonella Typhimurium, when used with a few conventional adjuvants, could not produce a balanced Th1 and Th2 immune response against Salmonella Typhimurium. Conclusions: In this study, we developed a novel nano-vaccine formulation composed of outer membrane proteins of Salmonella Typhimurium and calcium phosphate nanoparticles. The vaccine formulation was found to be safe and could elicit the desired Th1 and Th2 immune responses, as evidenced by humoral, cell-mediated, and protective immunity produced by the nano vaccine in poultry. Therefore, the present findings suggest that the CaP-Omp-Nps vaccine may be an efficient, safe, and cost-effective vaccine against Salmonella Typhimurium.

Pharmaceutics doi: 10.3390/pharmaceutics18060679

Authors: Ronan Pinto Nobrega dos Santos Dana Celeste Betancourt Roldan Muslum Guven Lucas Campana Leite Francisco Jacomine Madrid Furlan Gabriel Rocha Mariano da Silva Vitória Almeida Pessoa de Oliveira Carolline da Silva Capriglione Josie Pereira da Silva José Carlos Pinto Ismail Eş Tiago Albertini Balbino

RNA-based therapeutics are becoming increasingly important in oncology, particularly following the rapid development of mRNA technologies during the COVID-19 pandemic, but their success strongly depends on how efficiently they can be delivered to target cells. Microfluidic technologies have redefined the design and manufacturing of RNA-based nanocomplexes, as they enable precise control over physicochemical features that are critical for clinical translation in oncology. This review examines recent developments in microfluidic-assisted synthesis of RNA nanocarriers, with a focus on cancer applications. Through a detailed analysis of material systems, device architectures, and formulation strategies, we explore how laminar flow environments enable reproducible encapsulation, tunable particle size, and improved payload stability. We examine the microfluidic assembly of lipid nanoparticles and polymeric carriers for RNA delivery, highlighting strategies to enhance durability, bioavailability, and cellular uptake. Advancements in process optimization, including flow parameter refinement and inline monitoring, are discussed alongside the influence of device geometries on mixing dynamics and nucleation. Beyond formulation, we explore the integration of microfluidics with tumor-on-chip platforms to evaluate transport, penetration, and therapeutic response in physiologically relevant cancer models. By connecting technological innovation with preclinical application, this work outlines the trajectory toward next-generation, personalized RNA nanomedicines enabled by microfluidic precision.

Pharmaceutics doi: 10.3390/pharmaceutics18060680

Authors: Do-Hyub Kim Sung-Kwan Hwang Ji-Hyeon Yoon Dong Hee Na Young-Joon Park Yoon-Jee Chae Ji-Eun Chang Joo-Eun Kim

Background/Objectives: Oral delivery of semaglutide (Rybelsus) relies on sodium N-(8-[2-hydroxybenzoyl]amino)caprylate (SNAC) to enhance peptide absorption. However, formulation constraints and SNAC’s localized gastric mechanism have prompted the exploration of alternative enhancers. This study evaluated whether sodium caprate (C10), a well-characterized medium-chain fatty acid (MCFA), could achieve systemic exposure comparable to SNAC-based formulations when co-formulated in an immediate-release (IR) tablet. Methods: Preformulation studies assessed the physicochemical properties and buffering capacity of C10. Mechanistic feasibility was evaluated through Caco-2 transport studies and rat pharmacokinetic (PK) trials using aqueous suspensions, comparing the concentration-dependent effects of C10 and SNAC. Based on these findings, three IR tablet architectures (monolayer, bilayer, and dry compression-coated) were developed. The optimized formulation was evaluated in beagle dogs (14 mg semaglutide) and compared with the SNAC-based reference product. Results: C10 exhibited sufficient buffering capacity to neutralize acidic environments. In Caco-2 and rat PK studies, C10 enhanced semaglutide absorption in a concentration-dependent manner, yielding exposure levels equivalent to SNAC at matched doses. Among the tablet designs, the monolayer tablet showed the highest dissolution similarity (f2 = 67.8) to Rybelsus. In beagle dogs, the optimized monolayer formulation produced pharmacokinetic parameters, including Cmax, AUClast, and t1/2, that overlapped with those of the SNAC-based reference drug product under matched dosing conditions. Conclusions: These results demonstrate that C10 can effectively support oral semaglutide delivery when incorporated into a rationally designed IR tablet. The findings support the feasibility of MCFA-based permeation enhancer platforms as formulation alternatives to SNAC for oral peptide therapeutics.