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.

Pharmaceutics doi: 10.3390/pharmaceutics18060678

Authors: Juste Baranauskaite Ipek Ceken Asta Kubiliene Rima Jurate Gerbutaviciene Ebru Türköz Acar Cetin Tas

Objectives: this study aimed to develop and optimize an intranasal delivery system for Eplerenone (EPL) by incorporating Eplerenone-loaded liposomes (Elip) into an in situ gel system (Elip-GG). The goal was to prolong the residence time of the drug in the nasal cavity and ensure sustained release. Methods: Elip and unloaded liposomes were prepared using the thin-film hydration method. Key formulation variables such as encapsulation efficiency (EE%), mean particle size (MPS), polydispersity index (PDI), and zeta potential (ZP) were optimized. The Elip was then incorporated into a gellan gum (GG) in situ gel to form Elip-GG. The Elip-GG formulation was evaluated based on parameters such as pH, viscosity, rheological behavior, mechanical properties, and in vitro release. Results: the optimal Elip formulation exhibited an EE of 86.3%, a mean particle size of 86.56 nm, a PDI of 0.29, and a ZP of −29.86 mV. The cumulative drug release from the Elip-GG formulation exceeded 93% after 2.5 h. The Elip-GG formulation significantly increased the sustained release of Eplerenone when administered intranasally, offering a promising alternative to oral and parenteral delivery methods for hydrophilic antihypertensive drugs.

Pharmaceutics doi: 10.3390/pharmaceutics18060676

Authors: Cecilia Espíndola Ira Wirth Victoria Isabel Martín Eva Bernal José Antonio Lebrón María Luisa Moyá Rafael R. de la Haba Cristina Sánchez-Porro Antonio Ventosa Carmen M. Granados-Carrera Sara Molina Alvaro Hidalgo Manuel López-López Pilar López-Cornejo Francisco José Ostos

Background/Objectives: Flavonoids are low-molecular-weight polyphenolic compounds that are universally distributed in plants. They are a chemically varied group of secondary metabolites with a broad range of biological activity. The use of flavonoids is known to decrease the risk of many chronic diseases due to their radical scavenging, antioxidant, anti-inflammatory, anticarcinogenic, and antimutagenic properties. Limitations in the use of flavonoids include their low water solubility and poor stability, and therefore their low bioavailability. The encapsulation of flavonoids in different nanocarriers has helped to overcome this limitation. Taking this into account, in this work, the encapsulation of four flavones with several therapeutic applications—7-hydroxyflavone, 7,8-dihydroxyflavone, baicalein, and luteolin—in poly(lactic-co-glycolic) acid (PLGA)-derived polymeric nanoparticles (NPs) has been investigated. Methods: A physicochemical characterization of the NPs has been carried out using different techniques, including the evaluation of antioxidant and antimicrobial activities. Results: In all cases, the encapsulation efficiency of the four flavones in the prepared NPs was high (>90%), the zeta potential was about −31 mV, and the size was nanometric (~450 nm). The drug release from the nanoparticles was also studied, showing first-order kinetics. Statistical tools were applied to the release rate constants. The antioxidant activity and the in vitro antimicrobial activity of the free and flavone-loaded NPs were investigated, in the case of the latter using Gram-positive and Gram-negative bacteria. Results show that when the flavones are encapsulated, they retain their therapeutic properties. Conclusions: In summary, PLGA-based NPs not only prevent flavone degradation but also significantly boost solubility, ultimately optimizing bioavailability. Our results underscore these NPs as a promising platform for efficient flavone delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18060677

Authors: Vineet Mahajan Awanit Kumar Jeena Jacob Maged M. Costantine Lauren S. Richardson Rheanna Urrabaz-Garza Emmanuel Amabebe Ourlad Alzeus G. Tantengco Ananth Kumar Kammala Ramkumar Menon

Background: Low-dose aspirin (LDA) reduces preeclampsia (PE) risk by up to 40%, yet its molecular effects on chorion trophoblast cells (CTCs), a fetal membrane lineage at the feto-maternal interface, remain obscure. CTCs form a structural and immunoregulatory barrier whose dysfunction drives inflammation-associated membrane pathology in PE. Extracellular vesicles (EVs) released by CTCs may encode cellular stress and adaptation states, offering a molecular window into aspirin’s timing-dependent effects on PE risk modification. Methods: Human CTCs were challenged with cigarette smoke extract (CSE) to model oxidative stress-driven PE pathology. Two paradigms were tested: (1) prophylactic aspirin (4 and 40 µg/mL) before and/or flanking the CSE, and (2) therapeutic aspirin after the CSE challenge. The EVs were isolated via ultracentrifugation and size-exclusion chromatography, characterized by nanoparticle tracking and immunoblotting, and profiled by quantitative mass spectrometry. A network pathway analysis and machine learning biomarker selection defined the EV-encoded molecular states. Results: The CTC-derived EVs from the CSE-exposed cells carried a PE-like proteomic signature marked by suppressed VEGF/ECM remodeling, activated TNF-p53 apoptotic signaling, and heightened inflammation. Prophylactic low-dose aspirin shifted the EV cargo toward an EV-encoded signature consistent with preserved angiogenic potential (enrichment of VEGFA, COL1A1, and MMP14) and predicted attenuation of apoptotic and NF-κB pathway activity by an Ingenuity Pathway Analysis. High-dose aspirin produced broad transcriptional suppression without an accompanying pro-angiogenic EV signature. Therapeutic (post-injury) aspirin partially attenuated the injury-associated EV cargo but did not restore the angiogenic EV signature. An exploratory machine learning analysis of EV proteomes identified a candidate prophylactic biomarker panel anchored by HSPA8, SERPINF2, COL4A1, and PLOD1, mapped to the predicted angiogenic recovery and redox-balance pathways. These EV cargo readouts represent the predicted molecular states and require functional validation before clinical interpretation. Conclusions: The CTC-derived EV proteomic signatures capture the dose- and timing-dependent aspirin effects in this in vitro CTC model, positioning the chorion as a candidate pharmacological “secondary responder” favoring cellular resilience over classical anti-inflammatory suppression. As an exploratory hypothesis-generating study, EV-based molecular profiling could provide a foundation for future investigations aimed at stratifying aspirin responders from non-responders, although clinical validation in maternal plasma cohorts will be required before any translational application.

Pharmaceutics doi: 10.3390/pharmaceutics18060675

Authors: Bryant Menke Charlene H. Choo Marc Ohlhausen Timothy Kaftan Nam Nguyen Lindsay Helget Alan Erickson Christopher D. Conrady Steven Yeh

Background/objective: Suprachoroidal triamcinolone acetonide (TA) was recently FDA-approved and is emerging as a new alternative to other local therapies for macular edema (ME) associated with noninfectious uveitis (NIU). The objective of this study is to evaluate the preliminary safety and efficacy of suprachoroidal TA in patients with refractory ME secondary to NIU. Methods: This was a retrospective review of a small cohort of patients with refractory ME secondary to NIU treated with suprachoroidal TA from November 2022 to October 2023. Results: Six eyes from five patients with refractory ME secondary to NIU were included in the study. The cohort included two females (40%), and the median age was 62 years (IQR = 8). Ophthalmic diagnoses included intermediate uveitis (n = 2; 40%), birdshot chorioretinopathy (n = 1; 20%), autoimmune retinopathy (n = 1; 20%), and panuveitis (n = 1; 20%). The median logMAR visual acuity was 0.7 (Snellen 20/100) at baseline and improved to 0.3 (Snellen 20/40) during follow-up visits at 1 month and 2–3 months. The median central subfield thickness (CST) was 690 μm at baseline and improved to 367.5 μm and 309 μm at the follow-up visits at 1 month and 2–3 months, respectively. The initial improvement in logMAR visual acuity and CST was less pronounced at follow-up visits at 6–7 months and 11–12 months. Conclusions: This study demonstrates the safety of suprachoroidal TA and efficacy signals, including improvement in visual acuity and ME at 3 months in patients with severe, refractory ME secondary to NIU.

Pharmaceutics doi: 10.3390/pharmaceutics18060674

Authors: Yonggyu Kang Jeongeun Kim Jisu Park Subin Lee Youngjin An Kwang Suk Lim Hyun-Ouk Kim

The tumor microenvironment (TME) acts as a major barrier to effective drug delivery and contributes to drug resistance in solid tumors. Hypoxia, acidosis, and elevated interstitial fluid pressure limit drug penetration, while cancer-associated fibroblasts and immunosuppressive cells promote survival signaling, drug efflux, and metabolic adaptation. Polymeric drug delivery systems offer a promising strategy to address these barriers because their structures can be precisely engineered and designed to respond to TME-specific stimuli. These properties enable controlled drug release at tumor sites and help improve therapeutic efficacy while reducing systemic limitations. This review discusses how physicochemical and cellular components of the TME contribute to drug resistance and how polymeric nanomedicines can be designed to overcome these barriers. In addition, it examines key challenges that limit clinical translation, including tumor heterogeneity, variable enhanced permeability and retention effects, manufacturing scalability, and regulatory requirements. Finally, this review highlights the future direction of polymer nanomedicine and focuses specifically on developing rational material design, enhancing preclinical models, and developing clinically appropriate strategies to combat TME-mediated drug resistance.

Pharmaceutics doi: 10.3390/pharmaceutics18060673

Authors: Yinqiu Wu Jun Zhang Liyong Zhang Wei Li Yanyan Xue Shengzhe Zhang Hua Dai

Post-traumatic osteoarthritis (PTOA) is a rapidly progressing joint disorder initiated by acute injury, characterized by persistent inflammation, chondrocyte dysfunction, and extracellular matrix (ECM) degradation. Despite its clinical burden, effective disease-modifying therapies are lacking. Increasing evidence suggests that the ubiquitin–proteasome system, particularly E3 ubiquitin ligases, plays a pivotal role in regulating key pathogenic pathways involved in PTOA and represents a potentially druggable regulatory axis. In this review, we provide a comprehensive overview of the emerging roles of E3 ubiquitin ligases in PTOA, highlighting their involvement in inflammatory signaling, chondrocyte fate regulation, and cartilage matrix remodeling. We further integrate the current findings into a unified framework, in which E3 ligases act as central regulatory nodes linking injury-induced molecular responses to chronic joint degeneration. Importantly, we emphasize the pharmacological and translational potential of targeting E3 ubiquitin ligases as a novel therapeutic strategy. Recent advances in small-molecule modulators, gene-based interventions, and proteolysis-targeting chimeras (PROTACs) highlight the druggability of this regulatory system and provide new opportunities for disease-modifying treatment in PTOA. We also discuss the current challenges, including context-dependent effects, limited PTOA-specific validation, and delivery barriers. Overall, this review provides a comprehensive and therapeutically oriented perspective on E3 ubiquitin ligases in PTOA and highlights their potential as promising targets for pharmacological intervention and disease-modifying therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18060672

Authors: Ayesha Bibi Feroze-Bakht Lupe Carolina Espinoza Lilian Sosa Mireya Zelaya Dagmar Gualotuña Campoverde Jorge Morillo-Poma Marcelle Silva-Abreu Délia Chaves Moreira dos Santos Antonio J. Braza Ana Cristina Calpena

Background/Objectives: Carprofen (CP) is a potent non-steroidal anti-inflammatory drug whose clinical use is limited by systemic adverse effects associated with oral administration. The aim of this study was to develop and evaluate a CP-loaded nanoemulsion (CP-NE) as a topical formulation for the management of post-surgical inflammation in veterinary applications. Methods: CP-NE was physicochemically characterized in terms of droplet size, polydispersity index, morphology, pH, rheological behavior, spreadability, and stability. Biopharmaceutical performance was assessed through in vitro drug release and ex vivo permeation studies using porcine ear skin. Safety was evaluated using in vitro cytotoxicity assays in HaCaT keratinocytes, histological analysis of ex vivo porcine skin, and assessment of biomechanical skin parameters in mice. Finally, anti-inflammatory efficacy was investigated in a murine model. Results: CP-NE showed a mean droplet size of approximately 140 nm, low polydispersity, spherical morphology, and Newtonian flow behavior with good spreadability. Stability studies confirmed the absence of significant physical destabilization and acceptable chemical stability under refrigerated and room temperature conditions. Release studies demonstrated sustained drug release, while permeation assays revealed low systemic exposure and high drug retention within the skin. Safety evaluations indicated good biocompatibility with no cytotoxicity, no histological alterations in skin tissue, and no alteration of the skin’s biomechanical properties in volunteers. In vivo efficacy studies showed that CP-NE significantly reduced post-surgical inflammation, promoting faster restoration of skin architecture and improved wound appearance. Conclusions: These findings suggest that CP-NE represents a promising topical delivery system for localized anti-inflammatory therapy following surgical procedures, offering significant potential for veterinary applications.

Pharmaceutics doi: 10.3390/pharmaceutics18060671

Authors: Maria V. Erokhina Alexander G. Masyutin Georgii V. Lisichkin Pavel G. Mingalev Gennadii A. Badun Larisa N. Lepekha Irina V. Bocharova Ekaterina K. Tarasova Atadzhan E. Ergeshov

Background: Pulmonary tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis). Drug-resistant TB remains a major public health challenge and calls for new approaches to drug development. Targeted delivery of antibacterial agents using nanoscale carriers represents one such approach. A decisive factor for efficient targeting is the judicious selection of the carrier platform. Methods: In the present study, diamond nanoparticles were evaluated as a prospective vehicle for conveying anti-TB drugs to lung cells. Conventional and analytical transmission electron microscopy were used to analyze the localization of the nanodiamonds (NDs) in the lungs of M. tuberculosis-infected mice 30 days after nanoparticle administration and 44 days post-infection. Results: The study shows that the NDs co-localize with M. tuberculosis in foamy macrophages of the lung, residing in the same cellular compartments—phagosomes/phagolysosomes and lipid droplets. These in vivo results demonstrate a high degree of macrophage-specific accumulation of NDs relative to M. tuberculosis. Conclusions: Consequently, NDs can be considered a promising carrier for targeted delivery of anti-TB therapeutics to the lungs during TB-induced inflammation.

Pharmaceutics doi: 10.3390/pharmaceutics18060670

Authors: Saurabh Tiwari Katarzyna Mądra-Gackowska Marcin Gackowski Nokeun Park Łukasz Szeleszczuk

Background: The blood–brain barrier (BBB), which restricts the brain penetration of most small molecules and almost all biologics, continues to be a significant hurdle in the development of drugs for the central nervous system (CNS). During early-stage screening, a reliable computational prediction of BBB permeability, typically expressed as log BB, can help reduce the experimental load. Methods: We provide a well-validated machine learning system created solely using the B3DB experimental database, which includes 7807 chemicals with BBB+/BBB− annotations and 1058 compounds with in vivo log BB values. Using the Mordred library, a carefully selected set of 40 two-dimensional chemical descriptors was calculated from SMILES notation without the use of artificial data augmentation. Stratified five-fold cross-validation was used to comprehensively benchmark the nine methods used in this study. Results: On a held-out test set (n = 212), gradient boosting produced the greatest regression performance, with R2 = 0.6043, RMSE = 0.4740 log units, and MAE = 0.3326, which is in line with the upper range recorded for experimental BBB datasets. On an internal test set (n = 1562), the corresponding classifier obtained an AUC-ROC of 0.9476 and a balanced accuracy of 0.8568; on an independent external validation set (n = 175), it achieved an AUC-ROC of 0.9137. Topological polar surface area was found by SHAP analysis to be the primary factor influencing BBB permeability, with lipophilicity and ionization-related characteristics being the second and third most important factors, respectively. Nonlinear relationships in accordance with accepted pharmacokinetic principles were validated using partial dependence analysis. Conclusion: This study provides a reliable technique for predicting BBB permeability in CNS drug discovery.

Pharmaceutics doi: 10.3390/pharmaceutics18060669

Authors: Nadezhda B. Rudometova Ksenia I. Ivanova Vladislav V. Fomenko Andrey P. Rudometov Lyubov A. Kisakova Denis N. Kisakov Elena V. Yakovleva Vladimir A. Yakovlev Kristina P. Makarova Danil I. Vakhitov Mariya B. Borgoyakova Ekaterina V. Starostina Boris N. Zaitsev Victoria R. Litvinova Stepan A. Pyankov Tatiana N. Ilyicheva Alexander A. Ilyichev Andrei S. Gudymo Vasiliy Yu. Marchenko Nariman F. Salakhutdinov Aleksandr P. Agafonov Larisa I. Karpenko

Background: Avian influenza is a critical zoonotic infection threatening both the poultry industry and global public health. While traditional intramuscular vaccines elicit systemic immunity, they often fail to provide robust local protection at mucosal surfaces. There is thus significant interest in the development of mucosal avian influenza vaccines administered via the intranasal route. However, in humans, this approach is significantly hampered by the availability of safe and effective adjuvants. Methods: This study investigated the immunogenicity of a modified recombinant influenza A/H5 hemagglutinin (rHA/H5-modif) formulated with Novochizol, a novel chitosan-derived delivery system, administered intranasally to laboratory animals. Results: Our results demonstrate that mucosal immunization with the rHA/H5-modif/Novochizol complex induces potent humoral (IgG and IgA) and cell-mediated immune responses. Crucially, the formulation provided 100% survival in mice following a lethal challenge with highly pathogenic avian influenza A/H5. Conclusions: These findings position the rHA/H5-modif/Novochizol complex as a promising candidate for next-generation mucosal vaccines, in particular against highly pathogenic avian influenza A/H5 subtype.

Pharmaceutics doi: 10.3390/pharmaceutics18060668

Authors: Jeong Min Lee Kyung Tae Kim Chung-Sung Lee Hee Sook Hwang

Pulmonary fibrosis (PF) is a progressive and often fatal interstitial lung disease for which the currently available pharmacological therapies remain largely limited to slowing disease progression rather than reversing established fibrosis. This limitation has stimulated increasing interest in innovative therapeutic platforms capable of modulating complex fibrotic pathways. In this context, exosomes—nanoscale extracellular vesicles—have emerged as promising cell-free nanocarriers due to their intrinsic biocompatibility, low immunogenicity, and ability to be engineered for targeted drug delivery. In this review, we provide a comprehensive overview of both natural and engineered exosome-based strategies for the diagnosis and treatment of pulmonary fibrosis. We summarize recent advances in exosome engineering, including ligand functionalization, glycoengineering, and therapeutic cargo loading, highlighting how these approaches may support the development of more targeted and potentially personalized nanotherapeutic strategies. We further discuss emerging hybrid delivery platforms, such as exosome–liposome chimeras and hydrogel-based depots, which may enhance pulmonary retention, improve therapeutic durability, and enable controlled drug release. Finally, we outline key challenges and opportunities for clinical translation, including large-scale manufacturing, regulatory considerations, and clinically relevant delivery routes such as inhalation-based administration. Collectively, this review provides a translational perspective on engineered exosomes as emerging nanotherapeutic platforms for pulmonary fibrosis.

Pharmaceutics doi: 10.3390/pharmaceutics18060667

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

Background: Hot-melt extrusion (HME) is a promising technology for the manufacturing of drug products; however, its application is limited by elevated thermal and shear stresses that may induce degradation of thermolabile active pharmaceutical ingredients. One of the approaches to reducing processing temperatures is the use of polymeric systems with tailored thermal and rheological properties. The aim of the study was to develop an approach for the design of polymeric systems exhibiting a transient plasticization window, enabling a reduction in melt viscosity and improved processability under low-temperature extrusion conditions, followed by the formation of a structurally coherent matrix upon cooling. Methods: The compatibility of the initial polymers was assessed using laser microinterferometry. Based on the obtained data, three- and four-component polymeric compositions were designed and prepared by hot-melt extrusion. The resulting materials were characterized by differential scanning calorimetry, melt rheology analysis, and storage stability assessment. Thermal and rheological data were used to iteratively optimize the polymeric systems. Results: A four-component polymeric system based on PVP K-29/32, PEG 400, PEG 1500, and HPC EF was developed, suitable for processing by hot-melt extrusion at 70 °C. The final system enabled formation of a homogeneous extrudate, exhibited reproducible rheological behavior, and remained stable in the solid-state during storage, with no evidence of cold flow. Conclusions: It was established that, in the design of polymeric systems for hot-melt extrusion, the key factor is not achieving the lowest possible glass transition temperature, but rather the design of a system in which viscosity is transiently reduced under processing conditions and followed by structural stabilization upon cooling. The proposed approach may be applied in the development of polymeric premixes for the preparation of dosage forms by hot-melt extrusion, including those incorporating thermolabile active pharmaceutical ingredients.

Pharmaceutics doi: 10.3390/pharmaceutics18060666

Authors: Badmaarag-Altai Chuluunbaatar Yujin Jeong Jieun Ok Yujin Song Jae Woon Son Ji-Hyun Kang Wonwoong Lee Kyung Hyun Min

Background/Objectives: Cannabis sativa is the source of cannabidiol (CBD), a non-intoxicating phytocannabinoid with analgesic and anti-inflammatory qualities that has demonstrated therapeutic potential in inflammatory skin conditions like dermatitis. However, low bioavailability and poor water solubility restrict its topical application. This study attempted to improve CBD solubility and transdermal delivery using an amorphous solid dispersion (ASD)-based hydrogel system. Methods: CBD was stabilized in its amorphous form using an ASD strategy and incorporated into a hydrogel matrix. The CBD-ASD hydrogel was characterized by particle size analysis, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), rheological assessment, swelling studies, and diffusion experiments using Franz cells. Biological evaluations included cytotoxicity testing in human dermal fibroblast (HDF) cells, wound-healing assays, RT-qPCR-based anti-inflammatory analysis, antioxidant activity (DPPH assay), and antibacterial testing against Staphylococcus aureus. Results: Physicochemical analyses confirmed successful amorphous dispersion of CBD within a stable hydrogel network. The formulation exhibited sustained drug release over 144 h, achieving 86.32% cumulative release with diffusion-controlled kinetics. Rheological and swelling properties demonstrated mechanical stability and hydration suitability for long-term topical application, while Franz diffusion studies confirmed effective transdermal permeation. The CBD-ASD hydrogel showed no cytotoxicity in HDF cells and significantly enhanced wound closure. It also downregulated pro-inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α). Additionally, the formulation demonstrated 65.63 ± 10.00% DPPH radical scavenging activity and over 99% antibacterial inhibition. Conclusions: The CBD-ASD hydrogel represents a stable, multifunctional delivery platform that overcomes CBD solubility limitations and enhances therapeutic efficacy for inflammatory skin diseases.

Pharmaceutics doi: 10.3390/pharmaceutics18060665

Authors: Junqiao Mi Annabelle Schoder Aili Sun Patrick Meybohm Malgorzata Burek

Background: Brain microvascular endothelial cells (BMECs) form the blood–brain barrier (BBB), a highly selective interface that restricts paracellular diffusion and regulates the transport of nutrients and drugs into the central nervous system via specialized transporters and receptors. Tight junction-associated protein 1 (Tjap1), also termed protein incorporated later into tight junctions (Pilt), has been localized to tight junctions (TJs) in epithelial cells and to the trans-Golgi network in fibroblasts; however, its expression, subcellular localization, and functional significance in BMECs are still unknown. Methods: We characterized Tjap1 subcellular localization in mouse and human BMEC cell lines as well as primary mouse BMECs by immunofluorescence with and without pharmacological Golgi disruption by treatment with Brefeldin A, Golgicide A or Pitstop 2. CRISPR/Cas9-mediated Tjap1 knockout cells were generated and examined with regard to their Golgi morphology using immunostaining. Tjap1 mRNA localization was examined by RNAscope in situ hybridization. Quantitative real-time PCR and Western blot was performed to assess the expression of BBB-associated efflux transporters, solute carrier transporters, and cellular receptors in control and Tjap1 knockout cells. Results: Tjap1 predominantly localized to the cis-Golgi compartment, co-localizing with Gm130 rather than Tgn38, and was absent from TJs in BMECs. Tjap1 knockout induced pronounced Golgi fragmentation BMECs. Importantly, Tjap1 knockout significantly downregulated mRNA-expression of Abcb1a, Abcb1b, Abcc4, Slc2a1, Slc7a1, Slc7a5 and Tfrc, while Abcg2 was upregulated. At the protein level, a decrease in the protein levels of Abcb1, Abcc4, Slc2a1, Slc7a1, and Tfrc was observed in Tjap1 knockout cEND cells. Conclusions: In BMECs, Tjap1 is a cis-Golgi-associated protein required for the structural integrity of the Golgi apparatus. Its deletion is associated with Golgi fragmentation and significant alterations in the mRNA and protein expression of drug transporters and receptors at the BBB. These findings identify Tjap1 as a candidate regulator of both Golgi architecture and the BBB transporter profile in vitro, with potential implications for modulating drug transport across the BBB.

Pharmaceutics doi: 10.3390/pharmaceutics18060664

Authors: Arielly R. R. Barreto Ana Paula C. Valente Edgar Schaeffer Vitor M. de Almeida Michelle F. Muzitano Thiago Barth Alessandra M. T. de Souza Bárbara de A. A. Vieira Alcides Monteiro da Silva Osvaldo A. Santos-Filho Patrick G. Steel Bartira Rossi-Bergmann

Background/Objectives: Human and canine leishmaniases are neglected diseases with limited therapeutic options. The nitrochalcone NAT22, a high-affinity inhibitor of the essential parasite enzyme tryparedoxin peroxidase (cTXNPx), has emerged as a promising antileishmanial candidate. Interestingly, NAT22 demonstrated superior efficacy when administered orally rather than intralesionally, suggesting a metabolism-driven activity enhancement. Since in vivo studies with chalcones have been conducted exclusively in mice, this study aimed to determine whether mice are suitable models for oral chalcone therapies for human and canine leishmaniases and to identify metabolites with potential antileishmanial activity. Methods: NAT22 hepatic metabolism was investigated using in silico prediction and in vitro liver microsomal assays from rats, mice, humans, and dogs. Metabolites were identified by LC-MS/MS and NMR, and docking studies were performed against cTXNPx. Results: In silico analysis predicted metabolism mainly by CYP1A2, CYP2A6, CYP2C8, and CYP3A4. Seven metabolites (M1–M7) were identified by LC-MS/MS and NMR in all species except mice, whose microsomes did not generate M6. Structural analyses indicated preservation of the α,β-enone system and nitro-substituted B ring in all metabolites. Docking studies showed that metabolites M2 and M4 displayed stronger predicted binding energies than NAT22. Conclusions: NAT22 undergoes hepatic phase I metabolism generating two metabolites with enhanced predicted interaction with cTXNPx. The similarity between human and canine metabolic profiles supports the translational relevance of oral NAT22 therapy in leishmaniasis, while metabolites M2 and M4 emerge as candidates for validation in local treatment of cutaneous leishmaniasis.

Pharmaceutics doi: 10.3390/pharmaceutics18060663

Authors: Antonio Shegani

Cancer remains one of the most demanding health challenges worldwide, not only because of its high incidence and mortality, but also because of its biological heterogeneity, capacity for metastatic dissemination and frequent evolution under therapeutic pressure [...]

Pharmaceutics doi: 10.3390/pharmaceutics18060657

Authors: Qingshi Wen Ke Zhang Li Huang Shuyang Zhou Hanjie Ying Pengpeng Yang

Background: Daptomycin, a lipopeptide antibiotic with critical clinical applications, presents a formidable crystallization challenge due to its high conformational flexibility and complex ionization equilibrium. Current literature lacks reports on single crystals or highly crystalline powders of this molecule. This study aims to elucidate the thermodynamic and kinetic mechanisms governing daptomycin solubility and crystallization to establish a rational screening pathway. Methods: In this study, the solubility of daptomycin was systematically measured across eight pure solvents using a static gravimetric method. Molecular-level insights were obtained by integrating experimental data with the Conductor-like Screening Model for Real Solvents (COSMO-RS) and molecular dynamics (MD) simulations. Results: Solubility trends correlated strongly with solvent electrostatic and hydrogen-bonding capabilities. MD simulations revealed that the solvent’s ability to modulate conformational diversity—quantified by the number of dominant conformational clusters—is the decisive factor governing crystallization. For instance, aqueous systems exhibited strong Coulombic stabilization (−1126.61 kJ/mol). Crucially, solvents like acetone restricted daptomycin to a limited number of conformers (12 clusters), significantly lowering the nucleation barrier and yielding crystalline powders with distinct PXRD peaks. Conversely, solvents like methanol induced high conformational diversity (53 clusters), resulting exclusively in amorphous precipitates. Conclusions: The “number of conformational clusters” serves as a robust descriptor for rapidly screening crystallization solvents, effectively bridging thermodynamics and kinetics. This strategy provides a rational, reduced-trial-and-error framework for crystallizing complex, flexible macromolecules with multiple dissociation sites, moving beyond traditional trial-and-error approaches.

Pharmaceutics doi: 10.3390/pharmaceutics18060662

Authors: Milana Bosanac Bojana Andrejić Višnjić Aleksandra Popović Marko Ljubković Nikola Martić Dejan Miljković Alena Stupar Biljana Cvetković

Background/Objectives: Doxorubicin-induced cardiotoxicity (DIC) is driven by oxidative stress and impaired oxidative phosphorylation (OXPHOS). Antioxidant properties of carotenoids in vivo depend on extraction, while their direct role in mitigating DIC remains undetermined. This study evaluated the cardioprotective potential of natural deep eutectic solvents (NADES)-derived pumpkin carotenoid extract in a rat model of DIC. Methods: NADES-derived pumpkin pulp extract was characterized by spectrophotometry and HPLC-DAD. Wistar rats (n = 30) were assigned to six groups: C (control), N (NADES, 1 mL, p.o.), P (extract, 900 µg/kg body weight/day of total carotenoids, p.o.), D (doxorubicin, four i.p. doses, 2 mg/kg), ND (NADES/doxorubicin) and PD (900 µg/kg body weight/day of total carotenoids/doxorubicin). The activity of antioxidant enzymes and mitochondrial respiration in saponin-permeabilized left ventricular fibers using a Clark-type oxygen electrode) was measured. Results: Compared to control, cardiac antioxidant enzyme activities, mitochondrial respiration in complex I–linked respiration, ADP-supported OXPHOS, maximal respiratory capacity, and complex II and IV-linked respiration were significantly impaired by doxorubicin and unaltered by NADES. Co-administration of the extract significantly improved antioxidant enzyme activities (GSH-Px: D group 45 vs. PD group 95 nmol/mg proteins; GR: D group 95 vs. PD group 145 nmol/mg proteins; GST: D group 15 vs. PD group 22 nmol/mg proteins; SOD: D group 9 vs. PD group 17 U/mg proteins) and attenuated mitochondrial respiratory dysfunction compared with doxorubicin-treated group, indicating partial preservation of electron transport system capacity. Conclusions: Despite limitations of the study (single sex, single dose), results suggest NADES-based carotenoid extracts have cardioprotective properties in DIC by enhancing antioxidant defenses and supporting mitochondrial respiration.

Pharmaceutics doi: 10.3390/pharmaceutics18060660

Authors: Yuxiang Gantai Ziyan Yang Yinshuang Chen Mengxi Chen Yu Hu Tingwei Ye Jiayu Xu Shenyue Zhou Yuanyuan Yu Yan Chen Mengmeng Wang Weitao Zhang Jianqing Ruan Haiyang Zhang Weipeng Wang

Background: Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide, with more than 90% patients dying from metastasis due to limited treatment options. Although miRNA-based therapeutics represent a promising strategy, their clinical application has been hindered by poor stability in vivo and the lack of efficient organ-specific delivery systems. Methods: In this study, we developed a lung-targeted lipid nanoparticle (LuT-LNP) platform for the delivery of a chemically modified miRNA, AM22, which demonstrated enhanced tumor-suppressive activity. By replacing cholesterol and helper lipids with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), the most abundant lipid in pulmonary surfactant, and systematically optimizing the ratios of ionizable and cationic lipids, we obtained a LuT-LNP formulation with superior lung tropism. Results: The resulting LuT-LNPs exhibited excellent stability, biocompatibility, and efficient encapsulation and protection of AM22. Both in vitro and in vivo, AM22-loaded LuT-LNP (AM22@LuT-LNP) significantly inhibited the proliferation and migration of CRC cells and markedly suppressed lung metastasis in a mouse model. Mechanistic studies revealed that AM22 acts by targeting Poly (ADP-ribose) polymerase 1 (PARP1), inducing DNA damage, and inhibiting the epithelial-mesenchymal transition (EMT) process. Conclusions: These findings established a lung-targeted delivery platform for miRNA-based therapy, offering a promising strategy for the treatment of colorectal cancer pulmonary metastasis (CRPM).

Pharmaceutics doi: 10.3390/pharmaceutics18060661

Authors: Wenjie Ning Lingyu Hang Yuye Xue Wenting Zha Run Wang Kailin Xue Jiantao Ning Jiankang Zhao Liqiang Wang Hailong Yuan

Objective: Self-assembled nanoparticles (SANs) naturally occurring in Traditional Chinese Medicine (TCM) decoctions are promising drug carriers due to their biocompatibility, but uncontrolled assembly often leads to poor stability, limiting transdermal permeability and industrial application. This study aimed to fabricate stable and uniform SANs from licorice by precisely regulating the controlled nanoprecipitation of its water- and alcohol-extracted components. The transdermal delivery efficiency and therapeutic efficacy of the SANs in the treatment of atopic dermatitis (AD) were evaluated. Methods: Licorice self-assembled nanoparticles (LD-SANs) were prepared by mixing water and ethanol extracts of licorice, followed by ethanol evaporation under reduced pressure to trigger nanoprecipitation. In vitro transdermal tests compared the delivery efficiency of six major bioactive compounds between LD-SANs and traditional licorice decoction (LD). The penetration mechanism was investigated via passive diffusion and cellular uptake studies. In an AD mouse model, the therapeutic effects and expression of tight junction (TJ) proteins (Occludin and Claudin-1) were assessed. Results: The average particle size of LD-SANs is 200 nm, and it is uniform and stable. LD-SANs significantly enhanced the delivery efficiency of all six bioactive compounds compared to LD. Mechanistic studies revealed a unique “dual-channel” penetration mechanism: the nanoscale size enabled passive diffusion through hair follicles, intercorneocyte lipid gaps, and skin appendages, while perifollicular antigen-presenting cells (APCs) actively recognized and internalized the nanoparticles, creating a cell-mediated active targeting route that collectively boosted skin accumulation. In the AD model, LD-SANs promoted the expression of Occludin and Claudin-1 in the epidermal granular layer, reinforcing intercellular barrier integrity. Conclusions: By combining “efficient penetration” and “barrier repair”, LD-SANs demonstrated notable therapeutic efficacy in AD. This work transforms a traditional decoction into a well-characterized, high-performance nanomedicine and offers a novel strategy for developing TCM-based transdermal delivery systems.

Pharmaceutics doi: 10.3390/pharmaceutics18060659

Authors: Xiaoxiao Qiu Yilixiati Wusiman Nazhakaiti Yusufujiang Dilihuma Dilimulati Alhar Baishan Yipaerguli Paerhati Alifeiye Aikebaier Wenting Zhou

Plant-derived extracellular vesicles (PDEVs) are a novel category of natural nanocarriers with widespread availability, low immunogenicity, high biocompatibility, and inherent pharmacological activity. These features underscore their value as dual-function systems capable of serving as both carriers and bioactive agents. Unlike previous reviews that focused primarily on disease-specific applications or on individual engineering techniques, this review established a conceptual framework integrating three interconnected dimensions: (i) engineering strategies that address the inherent limitations of PDEVs (targeting, stability, loading efficiency); (ii) the carrier-performance-synergy paradigm linking PDEV composition to therapeutic outcomes; and (iii) gel-composite design principles that transform local retention into a controllable delivery platform. This review delves into various engineering methodologies, including targeted modification, enhanced stability, and optimized drug loading, while elucidating the performance characteristics of PDEVs as drug carriers, focusing on their protective, targeting, and controlled-release properties. It notably investigates the synergistic interactions between the intrinsic bioactivity of PDEVs and the drugs they deliver. Furthermore, this review highlights advanced applications of PDEV gel composites in localized drug delivery, specifically emphasizing their clinical potential for treating dermatological conditions. Finally, it highlights the current challenges faced by PDEVs and anticipates future research directions, such as synthetic biology, multi-omics analysis, and clinical translation. This review provides a theoretical framework for the rational design and clinical translation of PDEVs. It thereby promotes their innovative development in precision nanomedicine.

Pharmaceutics doi: 10.3390/pharmaceutics18060658

Authors: Zoltán Fekécs Krisztián Pajer Tamás Bellák Dénes Török Zsuzsanna Táncos Csilla Nemes László Gál Julianna Kobolák András Dinnyés Antal Nógrádi

Background/Objectives: Spinal cord injury (SCI) causes permanent tissue damage and severely limits functional recovery. We have previously shown that grafted neuroectodermal stem cells (NE-GFP-4C) promote regeneration and motor improvement of the injured cord through the release of specific factors, including Glial cell line-Derived Neurotrophic Factor, interleukin-6, interleukin-10 and Macrophage Inflammatory Protein-1α. Here, we examined whether various delivery routes for these factors could reproduce the effects of stem cell transplantation. Methods: Specifically, these factors were administered either in soluble form via osmotic pumps or via genetically modified fibroblasts. NE-GFP-4C cells served as a positive control, while saline and non-transfected fibroblasts were used as controls. Functional and morphological analyses were conducted to assess the therapeutic impact of the secretome treatment. Results: Both secretome-treated groups and stem-cell-grafted animals showed significantly improved motor outcomes, reduced lesion volumes, enhanced axonal regeneration and decreased astrocytic and microglial activation. Conclusions: These findings suggest that secretome-based delivery of selected factors is a minimally invasive and effective therapeutic strategy that can mimic the regenerative benefits of stem cell transplantation in SCI.

Pharmaceutics doi: 10.3390/pharmaceutics18060656

Authors: Zhewen Dong Xinxin Dong He Wang Yujie Pan Meiqi Yang Sihan Zhao Wanxian Deng Mengshan Han Tiantian Ye Shujun Wang

Background/Objectives: Insomnia severely impairs quality of life. Oral melatonin (MEL) suffers from poor brain delivery. Intranasal administration bypasses the blood–brain barrier, but rapid mucociliary clearance shortens drug retention, and MEL poor water solubility limits its nasal dissolution. Traditional in situ gels have “gelation-first, spreading-second” defects, causing uneven distribution. Herein, we developed a two-step sequential ion-triggered in situ gel combined with MEL liposomes (MEL-Lips-Gel) to enhance solubility, achieve instant uniform coating, and prolong retention for efficient nose-to-brain delivery. Methods: MEL-Lips were dispersed in alginate (first component) and calcium gluconate served as the second component. After sequential spray, the two components mix and form an ion-crosslinked gel. Rheology, in vivo fluorescence imaging, in vitro release, open-field/sucrose preference tests, and H&E staining were performed. Results: MEL-Lips showed uniform size and good encapsulation. The sequential system achieved instant widespread spreading and rapid gelation, significantly prolonged nasal retention, enabled sustained brain delivery, and reversed insomnia-induced hyperactivity and anxiety-like behaviors more effectively than oral MEL, intranasal MEL solution, liposomes alone, or non-liposomal gel, with good nasal safety. Conclusions: This sequential ion-triggered liposome-in-gel strategy synergistically overcomes rapid clearance (via gel) and poor solubility (via liposomes), enhancing nose-to-brain delivery of melatonin and providing a promising platform for insomnia therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18060655

Authors: Arunraj Tharamelveliyil Rajendran Ashwini Prabhu Ashwini Madhava Anoop Narayanan Vadakkepushpakath

Background: Glioblastoma (GBM) remains a lethal malignancy due to temozolomide (TMZ) resistance and limited drug penetration across the blood–brain barrier, largely driven by hyperactive DNA damage repair mechanisms such as poly (ADP-ribose) polymerase (PARP). To address these challenges, we developed folic acid-targeted PLGA–zein hybrid core–shell nanoparticles for the codelivery of the alkylating agent TMZ and the natural PARP inhibitor Ellagic acid (FA-TMZ/EA-PZ-CS NPs), thereby enabling simultaneous enhancement of drug delivery and suppression of chemoresistance pathways. Methods and Results: The dual-drug nanoplatform was fabricated using a double-emulsion solvent evaporation method and functionalized via EDC/NHS-mediated folic acid conjugation to promote receptor-mediated uptake. Physicochemical characterisation confirmed uniform spherical morphology, high colloidal stability, efficient drug encapsulation, and sustained biphasic drug release consistent with a core–shell diffusion mechanism. In LN229 glioblastoma cells, folic acid conjugation significantly enhanced cellular internalisation and cytotoxic efficacy compared to free drugs and non-targeted nanoparticles. Combination index analysis revealed strong synergism between TMZ and ellagic acid, resulting in markedly reduced IC50 values. Mechanistic studies demonstrated apoptosis induction, increased DNA damage, inhibition of cell migration at sub-cytotoxic concentrations, and downregulation of PARP gene expression. Conclusion: Overall, this study establishes a targeted core–shell nanotherapeutic strategy that integrates chemotherapy with DNA repair inhibition to overcome TMZ resistance, offering a mechanistically sound strategy that serves as a foundational framework for future translational research.

Pharmaceutics doi: 10.3390/pharmaceutics18060654

Authors: Amelia-Naomi Sabo Charlotte Nazon Catherine Paillard Véronique Kemmel

Background/Objectives: Hydroxyurea (HU), a cornerstone treatment for sickle cell disease (SCD), exhibits marked interindividual pharmacokinetic/pharmacodynamic (PK/PD) variability that remains poorly understood. This study aimed to establish a population PK model using OPTIMDREP randomized trial data (NCT06464458), quantify parameter variability, and identify covariates influencing HU PK and hematological response. Methods: Plasma sampling data from 22 SCD patients (20 pediatric and 2 adult patients; median age: 11.2 years [range: 2.5–35.7]) on once-daily oral HU underwent a non-compartmental analysis (NCA) followed by nonlinear mixed-effects modeling (MonolixSuite®). PK variability covariates and PK/PD correlations with the mean corpuscular volume (MCV), reticulocytes, fetal hemoglobin percentage (HbF%) and neutrophils were investigated. Results: NCA identified two absorption phenotypes (rapid/slow), with higher maximum concentration values observed for rapid (36.5 ± 18.8 mg/L) compared to slow (22.3 ± 8.4 mg/L) (p = 0.0013) profiles but not statistically different total exposures, apparent clearances (Cl/F) or volumes of distribution (Vd/F). The population approach identified the one-compartment model (first-order absorption and linear elimination) and confirmed the absorption phenotype as the key absorption rate (ka) covariate (9.93 vs. 1.36 h−1), explaining half of the ka interindividual variability (IIV). The median-normalized body weight was retained for both the Cl/F and Vd/F, as it significantly reduced the objective function value. No hematological parameter was correlated to PK parameters. However, rapid absorbers showed a superior response on the MCV (Δ = 9.1 fL, p < 0.0001), reticulocytes (Δ = −42.2 G/L, p < 0.01), and HbF% trend (Δ = 2.8%, p = 0.0835) but not on neutrophil counts (p = 0.8757). Conclusions: The absorption phenotype, a novel covariate explaining half of the ka IIV, predicts a superior erythropoietic response without toxicity in SCD patients. These findings support absorption phenotype integration into PK-guided dosing algorithms to optimize early-response biomarkers and personalize HU therapy.

Pharmaceutics doi: 10.3390/pharmaceutics18060653

Authors: Javiera Medina Thamara Hidalgo Fabián Martínez María Elena Gamboa-Arancibia Néstor Gutiérrez-Sánchez Sebastián Miranda-Rojas Alexander Gamboa

Background/Objectives: Amlodipine is an antihypertensive agent characterized by low aqueous solubility and variable oral bioavailability. This study aimed to formulate and characterize amlodipine–alginate nanoplexes and to incorporate the optimized system into an oral film dosage form. Methods: Nanoplexes were prepared via ionic complexation employing alginates (ALG) with diverse physicochemical properties, including low (LV) and medium (MV)-viscosity grades, as well as alginates with varying M/G ratios. The nanoplexes were thoroughly characterized employing a comprehensive set of analytical techniques. In addition, intermolecular interactions were examined using computational simulation studies. Results: The nanoplexes demonstrated high encapsulation efficiencies (>80%), with MV alginate yielding particles with greater drug loading but larger mean diameters compared with that prepared using LV alginate. Computational simulation studies revealed favorable interaction energies between the drug and the polyelectrolyte, particularly within microenvironments enriched in guluronic acid–rich repeat regions. These interactions were corroborated by infrared spectroscopy, while differential scanning calorimetry and X-ray diffraction analysis confirmed the amorphous solid state of amlodipine within the nanoplexes. Dissolution studies demonstrated an inverse relationship between alginate viscosity and drug release rate, with formulations based on LV alginate exhibiting rapid drug release. The final hydroxypropylmethylcellulose film incorporating ALG-MV nanoplexes exhibited adequate mechanical integrity and achieved approximately 95% drug release within 30 min. Conclusions: The developed film presenting a viable approach to enhance the delivery of amlodipine. Overall, this approach constitutes a significant advancement in the delivery of poorly soluble drugs through the integration of nanostructured systems with flexible oral film platforms.

Pharmaceutics doi: 10.3390/pharmaceutics18060652

Authors: Maša Sinreih Tisnikar Alja Zottel Katja Kristan Tea Lanišnik Rižner

Background/Objectives: Spinal muscular atrophy (SMA) is a rare genetic disorder caused by mutations or deletions in SMN1, resulting in the loss of SMN protein and severe neuromuscular consequences. Nusinersen, an antisense oligonucleotide that promotes full-length SMN2 transcript formation, has significantly improved SMA outcomes. However, standardized in vitro procedures for evaluating nusinersen efficacy remain limited. This study aimed to optimize in vitro efficacy assessment of nusinersen across two human cellular models. Methods: Experiments were performed using HEK293 cells and the SMA patient-derived fibroblast line GM03813. Transfection conditions were optimized for each model. In HEK293 cells, several seeding densities were evaluated for nucleofection, while in GM03813 fibroblasts, multiple transfection reagents and protocols were tested. Nusinersen activity was quantified at the transcript and protein levels, and dose–response curves were generated to determine EC50 values. Results: In HEK293 cells, a higher seeding density (1 × 106 cells) yielded the most efficient nucleofection. In GM03813 fibroblasts, Lipofectamine 3000 outperformed the other transfection reagents tested. Nusinersen exhibited dose-dependent effects in both models. The EC50 for transcript induction in HEK293 cells was 293 nM, whereas in GM03813 fibroblasts the EC50 was 10 nM, demonstrating substantial model-dependent differences in response. Conclusions: This study establishes optimized conditions for in vitro efficacy assessment of nusinersen in HEK293 and GM03813 cellular models. These protocols provide a robust and reproducible framework for evaluating nusinersen and can be readily applied to other antisense oligonucleotides designed to correct SMN2 splicing.

Pharmaceutics doi: 10.3390/pharmaceutics18060651

Authors: Caterina Nela Dumitru Alina Oana Dumitru Larisa Goroftei Elena Niculet Mariana Daniela Ignat Liliana Baroiu Aurel Nechita Gabriela Balan

Background: The gut microbiota constitutes a metabolically active “second genome” that profoundly modulates drug pharmacokinetics, pharmacodynamics, and adverse reaction profiles. Beyond antibiotics, widely prescribed non-antibiotic pharmacotherapies exert clinically relevant pharmacomicrobiomic effects with implications for therapeutic optimisation and pharmacovigilance. Methods: This narrative review, conducted following PRISMA 2020 reporting principles (without PROSPERO pre-registration), searched PubMed/MEDLINE, Scopus, Web of Science, and Cochrane Library (January 2015–December 2024) for evidence on proton pump inhibitors (PPIs), metformin, NSAIDs, statins, SGLT2 inhibitors, and oral iron. Evidence tables included clinical human studies with molecular microbiota characterisation (16S rRNA or shotgun metagenomics), ≥20 participants, and a control arm; preclinical data informed mechanistic synthesis. Results: Of 68 eligible studies, 20 met criteria for the evidence tables. PPIs significantly remodelled gut microbiota composition with enrichment of oral-origin taxa (“oralisation of the gut”), associating with Clostridioides difficile infection and SIBO. Metformin enriched Akkermansia muciniphila and butyrate producers, contributing causally to glycaemic efficacy. NSAIDs compromised barrier integrity, with synergistic dysbiosis under PPI co-prescription. Statins correlated with reduced prevalence of the dysbiotic Bact2 enterotype. SGLT2 inhibitor data remained discordant. Oral iron consistently enriched Enterobacteriaceae at the expense of beneficial commensals.

Pharmaceutics doi: 10.3390/pharmaceutics18060650

Authors: Minki Ha Seok-Beom Yong

Background/Objectives: Lipid nanoparticles (LNPs) are actively being studied as therapeutics and vaccines for various diseases. While LNPs can deliver nucleic acids, their efficiency is limited by the multi-step pathways involved in intracellular trafficking. Crucially, endosomal recycling-driven exocytosis acts as a major problem, rerouting LNPs away from the cytosol and thereby preventing efficient nucleic acid release. Upon entering the cell, LNPs are frequently expelled via endosomal recycling before delivering nucleic acids to cytosol. Previous studies reported that inhibition or deletion of Exo70, a component of the exocyst complex, leads to the accumulation of endosomes because of preventing endosomal recycling. In this study, we investigate the impact of Exo70 inhibition by endosidin-2 (ES-2), an Exo70 inhibitor, on LNP delivery efficiency. Methods: SM-102, cholesterol, DMG-PEG, and DSPC were dissolved in ethanol, while mRNA was dissolved in an aqueous phase to formulate LNPs. Co-treatment of ES-2 with LNPs was performed to evaluate its effect on mRNA delivery, and the resulting delivery efficiency was assessed both in vitro and in vivo. Results: Co-treatment of ES-2 with LNPs significantly enhanced mRNA delivery efficiency, resulting in up to a 4.06-fold increase in vitro and a 3.63-fold increase in vivo. Conclusions: Our findings demonstrate that suppression of Exo70 significantly enhances the mRNA delivery efficiency of LNPs, and this strategy could be applied for the development of mRNA therapeutics.

Pharmaceutics doi: 10.3390/pharmaceutics18060649

Authors: Congyi Zhang Haotian Wu Xiaotong Chen Wenze Yin Shizhuan Huang Dixiang Wen Xueting Song Xiaoyan Xu Changmei Zhang Sheng Tai

Background/Objectives: This study aimed to develop a novel tumor-associated macrophage (TAM)-targeting nanoplatform to improve the solubility and bioavailability of camptothecin (CPT) and achieve active targeted drug delivery for enhanced anti-tumor immunotherapy. Methods: We constructed a sialic acid-disulfide bond-camptothecin (SA-SS-CPT) nanowire system. Sialic acid was used as a targeting ligand to specifically recognize the overexpressed Siglec-E receptor on TAMs. Upon cellular internalization, the disulfide bond was designed to respond to intracellular glutathione (GSH), enabling controlled drug release. Results: The SA-SS-CPT nanowires significantly improved CPT solubility and enabled targeted delivery to TAMs. Following GSH-responsive cleavage and CPT release, the nanowires induced DNA damage in TAMs, activating the cGAS-STING signaling pathway. This promoted TAM polarization toward the M1 phenotype, enhanced pro-inflammatory and anti-tumor immune responses, and inhibited tumor immune escape. Furthermore, SA-SS-CPT synergistically improved the efficacy of PD-L1 blockade immunotherapy, remodeling the tumor immune microenvironment. Conclusions: The SA-SS-CPT nanoplatform effectively targets TAMs, repolarizes them to an anti-tumor M1 phenotype, and activates the cGAS-STING pathway. It shows strong potential for overcoming tumor immune escape and synergizing with PD-L1 checkpoint blockade to achieve significant tumor clearance.

Pharmaceutics doi: 10.3390/pharmaceutics18060648

Authors: Popat Mohite Abhijeet Puri Shubham Munde Nitin Ade Aarati Budar Anil Kumar Singh Deepanjan Datta Supachoke Mangmool Sudarshan Singh Chuda Chittasupho

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Pharmaceutics doi: 10.3390/pharmaceutics18060647

Authors: Wenndy Pantoja-Romero Alexis Lavín Flores Alejandro Lozada-Jerez MiaSara Perez-Salvá Fabiola Rosa-Suárez Orestes Quesada Magaly Martínez-Ferrer Gerardo Morell Brad R. Weiner

Background/Objectives: Andrographolide (ADG) is a plant-derived compound with promising anticancer properties, but its medical use is limited due to poor water solubility and low bioavailability. This study proposes developing a gold-based nanocomposite drug delivery system, using a simplified synthesis method, to improve ADG’s hydrophilicity and enhance its delivery efficiency. Methods: A one-step method was used to synthesize gold nanocomposites with carbon quantum dots (CBQDs) and doped CBQDs acting as reducing and stabilizing agents. These nanocomposites were then conjugated with ADG and thoroughly characterized using multiple structural and spectroscopic techniques such as X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), ultraviolet–visible spectroscopy (UV–Vis), transmission electron microscopy (TEM), Raman spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. Hydrophilicity enhancement was evaluated using NMR-based log P measurements. Biological assessment involved cell viability assays and confocal microscopy studies in PC3 prostate cancer cells, along with the morphological evaluation of human red blood cells. Results: XRD confirmed the formation of crystalline, face-centered cubic gold nanoparticles, while spectroscopic analyses verified successful nanocomposite formation and ADG conjugation. NMR results showed enhanced hydrophilicity of ADG. Biological tests demonstrated that the nanocomposites were compatible with cells. Conclusions: This study presents a straightforward strategy for synthesizing gold-based nanocomposites that enhance the hydrophilicity and delivery potential of andrographolide, supporting their applicability as nanocarrier platforms for anticancer drug delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18060646

Authors: Maryam Ranjpour Brion Frierson Rebekah Lynn Emerine Christian Jordan De Vera Krishna Sarva Melvin Earl Klegerman David Dugald McPherson Steven Idell Galina Florova Andrey Anatolievich Komissarov

Background: Pharmacological treatment under conditions of slow fibrinolysis/thrombolysis requires the targeted delivery of plasminogen-activating activity. Echogenic liposomal formulations (regular TELIP) of single-chain tissue plasminogen activator (sctPA), while possessing high affinity to fibrin, contain free/loosely bound sctPA. We hypothesized that removal of free sctPA, which competes with liposomes and plasmin for fibrin, enhances unique features of the TELIP. Methods: Optimized and regular TELIP were assessed for the distribution of active sctPA (loosely bound, tightly bound, encapsulated), stability, binding to fibrin, initiating fibrinolysis in vitro and ex vivo using a battery of biochemical methods. Results: One milligram of the regular TELIP consists of 2.0–5.0 × 109 echogenic liposomes (700–900 nm diameter). Non-specifically bound sctPA readily dissociates at the physiological ionic strength and pH. While up to 60% of sctPA in the regular TELIP is loosely bound with 6–15% encapsulated, and the rest is tightly bound to the liposomes; in the optimized TELIP, more than 80% of active sctPA is tightly bound with up to 40% of encapsulated. The latter is protected from high-molecular-weight ligands and could be released by an ultrasound pulse. Optimized TELIP shows low competition with plasmin for fibrin and effectively supports fibrinolysis in vitro and ex vivo. The optimized TELIP with maximal load of sctPA 3% (w/w) retains integrity at 37 °C for 5 h in vitro and up to 2 h ex vivo. Conclusions: The optimized TELIP is stable in vitro and ex vivo, does not interfere with fibrinolysis and retains a high level of encapsulated sctPA delivered precisely to the thrombus/fibrin clot.

Pharmaceutics doi: 10.3390/pharmaceutics18060645

Authors: Hideto To Mari Tomonari Makoto Myojin Fumiyasu Okazaki

Background: Irinotecan hydrochloride (CPT-11) is an important anticancer drug used in a wide range of regimens to treat colorectal and gastric cancers, and one of its severe side effects is delayed diarrhea. Therefore, based on known circadian variations in intestinal function and drug metabolism, we investigated whether CPT-11-induced delayed diarrhea may be attenuated by the time of dosing. Methods: When CPT-11 was administered to rats at 9:00 or 21:00, CPT-11-induced delayed diarrhea was assessed, and concentrations of CPT-11, its active metabolite SN-38, and SN-38 glucuronide (SN-38GL) in blood, intestinal tissues, and intestinal contents were measured. Results: The severity of diarrhea was significantly less in the 21:00 dosing group compared with the 9:00 dosing group. Blood SN-38 concentrations 8 h after the administration of CPT-11 were significantly higher in the 9:00 dosing group than in the 21:00 dosing group. SN-38, which exerts potent cytotoxic effects, circulates enterohepatically. When SN-38 is absorbed from the intestinal mucosa, intestinal tissues may be injured, resulting in delayed diarrhea. CPT-11 and SN-38 concentrations in intestinal tissues and contents 8 h after the administration of CPT-11 were significantly higher in the 9:00 dosing group than in the 21:00 dosing group at all measurement points. This was consistent with more severe CPT-11-induced delayed diarrhea in the 9:00 dosing group. Conclusions: Chronotherapy with CPT-11 may reduce CPT-11-induced delayed diarrhea. These differences in SN-38 concentrations in the intestinal tract at different dosing times may contribute to the time-dependent reduction in CPT-11-induced delayed diarrhea.

Pharmaceutics doi: 10.3390/pharmaceutics18060644

Authors: Shima Afrasiabi Mohammad Reza Karimi Sepideh Khoee Stefano Benedicenti Antonio Signore

Background/Objectives: Metabolic dormancy in biofilms leads to reduced drug efficacy in these communities. Different pharmacokinetics and adverse side effects complicate the simultaneous delivery of multiple drugs at appropriate concentrations to the infection site. This study aimed to develop chitosan-coated mesoporous silica nanoparticles loaded with curcumin and amphotericin B (CS@MSNs-Cur-AmB) and to evaluate their antibiofilm activity combined with antimicrobial photodynamic therapy (PDT) against Streptococcus mutans and Candida albicans dual-species biofilms. Methods: CS@MSNs-Cur-AmB were developed. The structure and morphology of the nanoparticles were evaluated using Fourier transform-infrared spectroscopy (FTIR), zeta potential, field emission scanning electron microscopy (FESEM), and thermogravimetric analysis (TGA). Cytotoxicity toward human gingival fibroblasts was assessed. Colony-forming units per milliliter (CFU/mL) were determined. The metabolic activity of biofilm-forming cells was measured using the tetrazolium (MTT) assay. Results: Physicochemical analyses confirmed the synthesis of CS@MSNs-Cur-AmB, revealing a particle size of 228 nm and thermal stability up to 600 °C. Cytotoxicity assays showed that CS@MSNs-Cur-AmB exhibited good biocompatibility (>90%). CS@MSNs-Cur-AmB improved antimicrobial activity, which was further enhanced by blue light-emitting diode (LED) irradiation. CS@MSNs-Cur-AmB under LED irradiation showed the strongest effect, reducing metabolic activity to 27.74 ± 4.08% (1 W/cm2, 1 min), p < 0.001). Conclusions: Formulating two drugs in nanocarrier systems may improve therapeutic efficacy by increasing local concentration and reducing systemic exposure. This offers an effective strategy for combating oral biofilms.

Pharmaceutics doi: 10.3390/pharmaceutics18060642

Authors: Ana Luisa Pinto Magalhães Nayara Santana Peixoto Moura Janaína de Alcântara Lemos Carolina de Aguiar Ferreira Danyelle M. Townsend Juliana de Oliveira Silva Anna Eliza Maciel de Faria Mota Oliveira André Luis Branco de Barros

Background/Objectives: The convergence of traditional medicinal practices in Brazil’s vast biodiversity has fueled pharmaceutical interest in advancing plant-derived formulation. Copaiba (Copaifera spp.) and andiroba (Carapa guianensis) are central to both the economic landscape and healing traditions of the Amazon rainforest. Derivatives from these species have diverse applications, with their oils representing important raw materials for therapeutic use. However, the poor aqueous solubility of oils remains a major barrier to developing formulations with optimal bioavailability. Nanotechnology offers a strategic approach to address this limitation, as nanosystems improve stability, solubility, and biological performance. Methods: This narrative review compiles and analyzes contemporary literature on the chemical composition, physicochemical properties, and pharmacological activities of copaiba and andiroba oils, with emphasis on studies involving nanoformulations, aiming to overcome the solubility limitations of these oils. Results: Evidence from the literature indicates that nanoencapsulation enhances the anti-inflammatory, antimicrobial, and wound-healing activity of the oils’ main constituents, such as beta-caryophyllene and limonoids. However, inconsistencies in reported chemical composition and physicochemical properties across studies highlight the lack of standardized characterization and extraction methods, potentially hindering the development of reproducible nanosystems. Conclusions: Nanoencapsulation represents a promising strategy to improve the therapeutic potential of Amazonian oils. Nevertheless, further efforts are required to standardize methodologies and expand clinical studies to confirm the efficacy and safety of nanosystems derived from these natural products.

Pharmaceutics doi: 10.3390/pharmaceutics18060643

Authors: Marek Misiak Aleksandra Maciejowska Maciej Pałęga Rafał Burek Anita Gołda Michalina Dworak Beata Pawuła-Prgomet Karol Forysiński Tomasz Miłek

Background/Objectives: Chronic inflammation is a key factor in the development and progression of colorectal cancer (CRC). When COX-2 levels and PGE2 production increase, nonsteroidal anti-inflammatory drugs (NSAIDs), including aspirin (ASA) and selective COX-2 inhibitors, such as celecoxib and rofecoxib, are commonly employed. This paper presents the effect of anti-inflammatory drugs, primarilyNSAIDs, on the incidence of CRC. Methods: A comprehensive literature search (119 articles) was conducted with databases such as PubMed. During our research, we used keywords such as colorectal cancer (CRC), nonsteroidal anti-inflammatory drugs (NSAIDs), ASA, COX, precision oncology, and personalized medicine. Results: The development of CRC is primarily associated with chronic inflammation and the actions of COX-2 and prostaglandin E2 (PGE2), which promote cancer cell proliferation and angiogenesis. Anti-inflammatory drugs act by inhibiting the secretion of COX-1 and COX-2 enzymes, which leads to reduced PGE2 production and may limit tumor growth. Aspirin has the best-documented and studied anti-cancer effect; long-term use is associated with a reduced risk of CRC development and mortality through its anti-inflammatory and antiplatelet effects, thereby limiting metastasis. Particularly beneficial effects are observed in patients with mutations in the PIK3CA gene. Factors influencing the effectiveness of CRC treatment include molecular differences and tumor location. Conclusions: The future of CRC treatment and prevention lies in personalized medicine, which accounts for each patient’s genetic profile. Decisions regarding NSAIDs use and CRC prevention should consider the potential benefits and risks of side effects.

Pharmaceutics doi: 10.3390/pharmaceutics18060641

Authors: Naixuan Deng Yeqi Huang Yue Gao Hongluo Li Wenjing Wang Minjing Cheng Chuanbin Wu Xin Pan Ling Guo Junhuang Jiang Zhengwei Huang

Supramolecular inclusion complexes are widely used in drug delivery and other fields, with the advantages of controllable structures, high stability, excellent biocompatibility, and the ability to improve drug solubility and achieve controlled release. However, traditional screening methods rely on experimental trial and error, which suffer from long cycles, high costs, and low throughput, limiting research and development efficiency. In recent years, the development of artificial intelligence has provided new solutions for the screening of inclusion complexes. This paper systematically reviewed the core technological system of AI in the screening of inclusion complexes, focusing on two aspects: prediction and optimization of key properties and rational design of host molecules, summarizing their specific application progress. At the same time, we analyzed the current core challenges, including data scarcity, insufficient model interpretability, and limited generalization capabilities, and propose future development directions such as building standardized databases, integrating physicochemical principles (e.g., molecular mechanics and thermodynamics), and establishing closed-loop research and development platforms. This review aims to provide a systematic reference for the in-depth application of artificial intelligence in the field of supramolecular inclusion complexes.

Pharmaceutics doi: 10.3390/pharmaceutics18060640

Authors: Maria-Medana Drăgoi Florina-Diana Goldiș Sabina-Oana Vasii Daiana Colibășanu Liana Suciu Angela Caunii Lucreția Udrescu

Background: Chronic obstructive pulmonary disease (COPD) is commonly managed alongside multimorbidity, polypharmacy, recurrent treatment escalation, and older age, all of which increase vulnerability to drug–drug interactions (DDIs). We aimed to synthesize the main DDI domains relevant to COPD pharmacotherapy and to distinguish harmful DDIs from beneficial combination therapy and formal compatibility findings. Methods: We performed a narrative review using structured literature searches and citation tracking to evaluate COPD-related studies. We prioritized direct COPD-specific DDI evidence, while also including mechanistic, class-specific, and contextual studies when direct evidence was lacking. Retained evidence included observational cohorts, prescribing studies, pharmacokinetic trials, case reports, and systematic reviews. Results: The reviewed literature indicates that DDI vulnerability in COPD is driven less by isolated drug pairs than by overall regimen complexity, multimorbidity, aging, fragmented prescribing, and high-intensity treatment periods such as exacerbations, hospitalization, and discharge. Key DDI domains included cardiopulmonary co-treatment, QT-related vulnerability, and potential or clinically relevant interactions amplified during exacerbations. Inhaled therapies are not universally interaction-free, particularly with strong metabolic inhibitors. Psychotropics, frailty, dementia, and palliative care further increase clinical complexity. However, beneficial bronchodilator combinations and formal compatibility studies demonstrate that not all multidrug COPD regimens are harmful. Conclusions: In COPD, DDI assessment should focus on the full treatment regimen and not be limited to a set of iconic drug pairs. Clinicians must focus on exacerbation-related prescribing, QT-active drugs, theophylline exposure, psychotropic co-medication, and vulnerable subgroups such as older, frail, and palliative patients. Pharmacist-supported drug review, drug reconciliation, and selective deprescribing are key strategies for reducing clinically relevant DDI burden in COPD.

Pharmaceutics doi: 10.3390/pharmaceutics18060639

Authors: Mengmeng Guan Wanyi Zhou Haoran Qin Yi Xu Di Zhao Hui Xue Nan Hu

Objective: Posaconazole, a second-generation triazole antifungal used for the prevention or treatment of invasive fungal infections, has been shown to markedly increase tacrolimus exposure in vivo when co-administered, potentially leading to clinically significant adverse events. A physiologically based pharmacokinetic (PBPK) model was developed to predict tacrolimus–posaconazole interactions in renal transplant recipients with different CYP3A5 genotypes, to inform tacrolimus dose adjustment in clinical practice. Methods: First, to obtain the critical inhibition parameters, in vitro enzyme kinetic studies were conducted. Based on these data, a whole-body physiologically based pharmacokinetic (PBPK) model for TAC was developed and validated in PK-Sim. A published, validated posaconazole PBPK model was applied concurrently. Model performance was evaluated against published pharmacokinetic data in healthy volunteers receiving tacrolimus with posaconazole. A virtual Chinese renal transplant recipient was generated by incorporating population-specific physiological parameters, including CYP3A5 genotype-dependent enzyme expression. Results: In vitro experimental results demonstrated that POSA acts as a potent reversible competitive inhibitor of CYP3A4/5-mediated TAC metabolism. The tacrolimus PBPK model adequately captured pharmacokinetics across CYP3A5 genotypes, and tacrolimus pharmacokinetics during co-administration with posaconazole were also predicted. Compared with CYP3A5 expressers, nonexpressers showed greater variability in tacrolimus whole-blood concentrations and greater susceptibility to posaconazole-mediated interactions. The CYP3A5*3*3 genotype was associated with higher Cmax and AUC. Dose optimization simulations predicted that after 6–7 days of posaconazole co-administration, nonexpressers would require the reduction of tacrolimus dosing frequency from every 12 h to every 24 h to maintain trough concentrations within 8–15 ng/mL, whereas a 50% dose reduction was predicted to be optimal for expressers. Conclusions: A tacrolimus–posaconazole PBPK drug–drug interaction model was developed for the population of renal transplant recipients and used to simulate tacrolimus trough concentrations across CYP3A5 genotypes and dosing regimens, supporting genotype-informed co-administration in clinical practice.

Pharmaceutics doi: 10.3390/pharmaceutics18060638

Authors: Bo Han Yue Zhang Yangmin Wang Yue Shen Jinping Niu Shipo Li Yuxi Li Jingyu Wang Xingyuan Ma Wenyun Zheng

Cannabidiol (CBD), a major non-psychoactive phytocannabinoid, has attracted considerable attention owing to its broad therapeutic potential. Its anti-inflammatory, antimicrobial, and antitumor properties make it a promising candidate for the treatment of mucosa-associated diseases. However, the clinical translation of CBD is significantly hindered by its unfavorable physicochemical properties, particularly high lipophilicity and poor aqueous solubility, which result in low bioavailability. To overcome these limitations, the rational selection of administration routes in combination with advanced drug delivery systems tailored to disease pathophysiology is essential. Such strategies are critical for improving the stability of CBD, enhancing mucosal permeation, and enabling controlled and targeted release at diseased sites. Nevertheless, a systematic review focusing on these aspects is still lacking. This review first summarizes the relationship between CBD and the mucosal endocannabinoid system, together with its pharmacological effects. It then discusses the therapeutic potential of CBD in mucosal disorders of the digestive and respiratory systems. In addition, current administration routes and advanced delivery systems for CBD are reviewed to provide insights for future research and clinical translation. Finally, the remaining challenges associated with the clinical application of CBD and future development directions are discussed.

Pharmaceutics doi: 10.3390/pharmaceutics18060637

Authors: Shailvi Soni Terrick Andey

Background: Oral delivery of chemotherapeutic agents remains challenging due to gastrointestinal degradation, poor intestinal permeability, and extensive first-pass metabolism, which collectively limit bioavailability. Lipid-based drug delivery systems offer a promising strategy to overcome these barriers. This study aimed to develop a freeze-dried, solid-dispersible self-emulsifying drug delivery system (SEDDS) using a water-in-oil-in-water (w/o/w) double emulsion approach for the co-encapsulation of hydrophilic (doxorubicin) and lipophilic (ellipticine) agents to enhance oral delivery. Methods: Double-emulsion SEDDS were prepared via a two-stage emulsification process to enable compartmentalized drug loading within aqueous and oil phases. The formulations were freeze-dried to improve stability and storage. Physicochemical properties were characterized using dynamic light scattering for droplet size and polydispersity index (PDI), zeta potential analysis for colloidal stability, and differential scanning calorimetry for thermal behavior. Drug encapsulation efficiency was determined, and cellular uptake was evaluated in breast cancer cells using fluorescence microscopy. Results: Optimized SEDDS exhibited droplet sizes of 90–347 nm with low PDI values (0.005–0.336), indicating uniform and stable dispersions. Zeta potential values (−10.64 to 2.38 mV) supported colloidal stability, while freeze-dried formulations retained dispersion characteristics upon reconstitution over extended storage. Both drugs demonstrated high encapsulation efficiency (>97%), and thermal analysis confirmed the formation of stable amorphous systems. Fluorescence imaging revealed enhanced intracellular uptake of both agents. Conclusions: This study demonstrates that freeze-dried double-emulsion SEDDS enable efficient co-delivery of hydrophilic and lipophilic drugs, improving stability and cellular uptake. This platform shows strong potential for overcoming key barriers in oral chemotherapy and provides a promising strategy for combination drug delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18060636

Authors: Lara Marques Nuno Vale

Background/Objectives: Propranolol (PROP) is a non-selective β-blocker widely prescribed for cardiovascular and neurological disorders. Its pharmacokinetics (PK) are highly variable, and co-administration with omeprazole (OME), a CYP2C19 substrate and inhibitor, may alter systemic exposure. Herein, this study aimed to investigate factors influencing PROP PK variability and evaluate the effect of OME coadministration using physiologically based pharmacokinetic (PBPK) modeling and population PK (popPK) analysis. Methods: PBPK models for PROP and OME were developed and validated against published data. DDI simulations were conducted across clinically relevant dosing regimens. A two-period fixed-sequence virtual trial of 125 subjects was simulated with PROP alone and PROP combined with OME. Population PK (popPK) analysis was performed on simulated plasma concentration data to identify covariates affecting PROP disposition and quantify DDI magnitude. Results: PBPK models were successfully developed and validated. PROP disposition was best described by a two-compartment model with linear elimination. Health status was found to influence clearance, and body surface area (BSA) affected the central volume of distribution. Co-administration with OME increased PROP exposure, with larger effects in patients with renal impairment. Simulated plasma concentrations remained below established toxicity thresholds. Conclusions: Virtual clinical trials integrating PBPK and popPK modeling provide a robust approach to identifying key determinants of PK variability and DDI risk. Although these findings were not directly translated to clinical observations, this helps identify sources of PK variability in PROP treatment settings and factors that may intensify its interaction with OME, thereby supporting model-informed precision dosing to enhance safety and efficacy.

Pharmaceutics doi: 10.3390/pharmaceutics18060635

Authors: Reihaneh Manteghi Eduardo Veas

Background/Objectives: Pharmaceutical formulation involves designing a drug product by combining the properties of an active pharmaceutical ingredient (API) with suitable excipients and processing strategies to produce a safe, effective, and manufacturable dosage form. However, data in formulation science are often limited, expensive to generate, and frequently restricted by proprietary and confidentiality constraints. Interactive digital tools can support formulators during early drug product development by improving the structure, transparency, and efficiency of formulation decision-making. While the current system focuses on structured decision support, future extensions may incorporate machine-learning methods for recommendation and knowledge extraction. Methods: In this work, we developed the Formulation tool, an interactive decision-support and visualization system for formulation development based on a hierarchical formulation-strategy framework commonly used in pharmaceutical practice. The tool is designed to prioritize suitable formulation principles and associated processing routes, with oral solid formulation as the initial application domain. The evaluated scenarios also include pathway regions relevant to oral liquid formulations. Its modular architecture also makes it adaptable to other formulation scenarios. To assess practical applicability, the tool was evaluated in a structured expert study involving five expert users across six predefined formulation scenarios (n = 30 runs), covering three drugs under adult and pediatric conditions. Results: The tool showed agreement with the expected dosage-form families and overall formulation properties, with adult scenarios converging to oral solid regions and pediatric scenarios converging to oral liquid regions. At the downstream formulation-profile level, users converged either to the dominant expected pathway or to alternative feasible pathways within the same formulation region. Variability in full pathway completion was observed and was primarily associated with differences in user interaction behavior and exploratory usage patterns. The median task completion time was 113.5 s. Conclusions: In addition to organizing formulation knowledge, the Formulation tool records user interactions in a structured manner, which may support future learning from usage patterns. Because detailed downstream formulation constraints are often institution-specific and are typically not available in the public domain, the present evaluation focused on agreement at the dosage-form-family level and on overall formulation properties rather than on highly specialized constraint logic. The system is based on a constraint satisfaction problem (CSP) framework, which is well suited for modeling complex decision processes under explicit constraints. CSP has also been widely applied in intelligent scheduling systems, supporting its suitability for structured, constraint-rich decision-making tasks such as pharmaceutical formulation strategy development.

Pharmaceutics doi: 10.3390/pharmaceutics18060634

Authors: Yan Lin Xian Zhang Fulin Bi Guangji Wang Jin Yang

Background/Objectives: Despite the presumption of bioequivalence for BCS Class I drugs due to their high solubility and permeability, recent evidence indicates that those with rapid systemic elimination exhibit heightened vulnerability to Cmax non-equivalence, primarily attributable to intrasubject variability in gastrointestinal transit and absorption kinetics. It is well known that gastric emptying is a significant physiological-dependent factor. But, does the formulation affect gastric emptying? Methods: Using zidovudine as a model drug, formulations containing sodium carboxymethyl starch (CMS-Na), pregelatinized starch, hydroxypropyl methylcellulose (HPMC), and lactose were investigated for their effects on gastric emptying kinetics, and the impact of excipient-mediated gastric emptying prolongation on pharmacokinetic parameters was also evaluated. Results: Relative to AZT alone (Cmax = 13,350 ng/mL; gastric %ID = 11.3%), co-administration with CMS-Na, pregelatinized starch, or HPMC significantly prolonged gastric retention (%ID: 23.4%, 30.5%, and 40.8% at 22.5 min) and reduced Cmax in rats by 47.8%, 34.4%, and 35.1%, respectively, with no effect on intestinal permeability. Viscosity positively correlated with gastric emptying delay. Conclusions: Our rat findings provide new possible mechanistic evidence that certain viscosity-modifying excipients can delay gastric emptying and reduce Cmax of zidovudine, a rapidly eliminated BCS Class I drug, with potential implications for biowaiver risk assessment. Gastric emptying is not only a physiological-dependent variation but also, in cases where common excipients may significantly delay gastric emptying, a formulation-dependent rate-limiting step. For such drugs, excipient-induced gastric emptying delay poses an underappreciated risk to the biowaiver approach, necessitating more prudent regulatory assessment that encompasses the dynamic interplay among sequential rate processes governing drug disposition.

Pharmaceutics doi: 10.3390/pharmaceutics18060633

Authors: Andrea Fernández Yasmín Cura-Cuevas Susana Rojo-Tolosa José María Gálvez-Navas Encarnación González-Flores Cristina Pérez-Ramírez Alberto Jiménez-Morales

Background/Objectives: Breast cancer (BC) is a highly prevalent neoplasm worldwide. Despite the wide range of therapeutic options currently available, it remains the leading cause of cancer-related mortality among women. Capecitabine, a prodrug of 5-fluorouracil (5-FU), is widely used in the treatment of advanced BC. However, despite its efficacy, capecitabine exhibits considerable interindividual variability in therapeutic response. This study aimed to evaluate the effect of single-nucleotide polymorphisms (SNPs) in genes involved in capecitabine bioactivation on progression-free survival (PFS) in patients with BC. Methods: An ambispective cohort study was conducted. Four relevant SNPs in the CES1, CDA, and TYMP genes were analyzed in 85 Caucasian patients with BC using real-time polymerase chain reaction (PCR) with TaqMan® probes. Results: A significant association was observed between shorter PFS and the GA genotype of the CES1 rs71647871 SNP (p = 0.010; HR = 7.46; 95% CI = 1.24–122.52), as well as with the TT genotype of the CDA rs602950 SNP (p = 0.009; HR = 3.50; 95% CI = 1.36–9.03). Conclusions: These findings suggest that CES1 rs71647871 and CDA rs602950 may serve as predictive biomarkers of capecitabine effectiveness in patients with BC. Further studies involving larger cohorts are needed to validate these findings and generate additional evidence to support their potential implementation in clinical practice.

Pharmaceutics doi: 10.3390/pharmaceutics18050632

Authors: Muhammad Sohail Khan Imran Zafar Dayeon Ham Ki Sung Kang Il-Ho Park

Carbon dots (CDs), including carbon quantum dots (CQDs), are ultra-small carbon-based nanomaterials, typically below 10 nm, with tunable photoluminescence, high aqueous dispersibility, favorable biocompatibility, low toxicity, and abundant surface functional groups. These properties make CDs promising multifunctional platforms for nanomedicine, particularly in bioimaging, biosensing, targeted drug/gene delivery, photodynamic therapy (PDT), photothermal therapy (PTT), antimicrobial treatment, and theranostic applications. This review critically examines recent advances in CD fabrication, including top-down, bottom-up, green biomass-derived, microwave-assisted, hydrothermal, and emerging hybrid strategies, with emphasis on how precursor selection, heteroatom doping, surface passivation, and polymer/ligand functionalization regulate optical performance, biological interaction, and therapeutic efficiency. The review discusses structural classification, including CQDs, graphene quantum dots (GQDs), carbon nanodots, and carbonized polymer dots (CPDs), together with major characterization approaches such as ultraviolet–visible (UV–Vis) spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy, and high-resolution transmission electron microscopy (HRTEM). Particular attention is given to red/near-infrared (NIR) emission, renal clearance, drug-loading behavior, reactive oxygen species (ROS) generation, toxicity mechanisms, biodistribution, and long-term biosafety. This review also highlights key translational barriers, including batch-to-batch variability, limited standardization, scalable manufacturing, regulatory uncertainty, and incomplete pharmacokinetic evaluation. It considers artificial intelligence (AI) and machine learning (ML) as emerging tools for reproducible CD design. CDs represent versatile and clinically promising nanoplatforms, but their translation requires standardized synthesis, rigorous safety assessment, and application-specific regulatory validation.

Pharmaceutics doi: 10.3390/pharmaceutics18050631

Authors: Alina Diana Panainte Cătălina Anișoara Peptu Andreea Crețeanu Nela Bibire Isabella Nacu Liliana Vereștiuc Eliza Grațiela Popa Larisa Păduraru Liliana Mititelu Tartau Radu Dănilă Tudor Bibire Catalina Natalia Yilmaz

Hyaluronic acid (HA) hydrogels have attracted increasing interest for biomedical applications due to their tunable properties and biocompatibility. Methods: In this study, hyaluronic acid HA-based hydrogels were developed using two distinct crosslinking strategies: physical crosslinking through poly(vinyl alcohol) (PVA) incorporation and covalent crosslinking via DCC/NHS-mediated reactions. Piroxicam (Px) was included as a model drug to evaluate the drug delivery potential of the resulting systems. The hydrogels were characterized in terms of morphology, swelling behaviour, adhesion, enzymatic degradation, drug release, and in vitro cytocompatibility. Results: The results indicate that formulation parameters significantly influence the overall performance of the systems. PVA-containing hydrogels exhibited higher swelling capacity and improved adhesive properties, while covalently crosslinked networks showed reduced swelling and enhanced structural stability and resistance to enzymatic degradation. Drug release profiles were dependent on network structure, with more compact systems displaying slower release behaviour. In vitro assays suggested that the developed hydrogels are cytocompatible and that drug incorporation influences both release kinetics and cellular response. However, it should be noted that the biological evaluation was performed under simplified in vitro conditions, which primarily reflect specific aspects such as cell viability and migration. Conclusions: This study provides a comparative analysis of physical and covalent crosslinking strategies within a HA platform and highlights how formulation variables influence key physicochemical and biological properties. These findings contribute to the rational design of HA-based hydrogels, although further studies are required to establish their performance in more complex biological environments.

Pharmaceutics doi: 10.3390/pharmaceutics18050630

Authors: Candela Canton Laura Ceballos Lucila Canton Laura Moreno Paula Domínguez Luis Alvarez Carlos Lanusse

Background/Objectives: Combination of antiparasitic drugs with different mechanisms of action has been suggested as an effective strategy to delay the development of parasite resistance. Considering the need to understand the pharmacological basis of drug combinations, the current study evaluated the potential pharmacokinetic (PK) interactions and the clinical efficacy (pharmacodynamic response) occurring after the subcutaneous administration of ivermectin (IVM) and levamisole (LEV), administered either as single treatments or concurrently to different groups of parasitized calves on three commercial farms (A, B and C). Methods: Forty-five (45) male calves naturally infected with gastrointestinal nematodes were randomly allocated into three groups (n = 15): IVM, treated with IVM by subcutaneous injection (0.2 mg/kg); LEV, treated subcutaneously with LEV (8 mg/kg); IVM + LEV, simultaneously treated with IVM and LEV (two subcutaneous injections at the same dose rates). Seven animals from each treated group (farm C) were randomly selected to perform the PK study. Drug concentrations were measured by HPLC. The therapeutic response (efficacy) was determined at 14 days after treatment by the fecal egg reduction test. Results: The mean area under the concentration vs time curve (AUC) for IVM obtained after administration of IVM alone (274 ± 65.1 ng.d/mL) was similar to that obtained when IVM was co-administered with LEV (295 ± 111 ng.d/mL). Likewise, mean LEV AUC values were similar after LEV administration alone (8.90 ± 2.69 µg.h/mL) or combined with IVM (9.11 ± 1.82 µg.h/mL). No adverse PK interactions were observed after the combined treatment, with similar PK parameters (p > 0.05) obtained between the single-drug and the combination-based strategies. On farm A, the overall fecal egg reductions were 38% (IVM), 99% (LEV) and 100% (IVM + LEV). While Cooperia spp. and Haemonchus spp. showed reduced susceptibility to IVM treatment, LEV demonstrated high efficacy against both genera, with only a minimal proportion of Haemonchus spp. remaining after treatment. Similarly, total fecal egg reductions were 42% (IVM), 99% (LEV) and 100% (IVM + LEV) on farm B, and 54% (IVM), 99% (LEV) and 100% (IVM + LEV) on farm C. On those farms, IVM was ineffective against Cooperia spp. and/or Haemonchus spp., while LEV failed to control Ostertagia spp. Remarkably, the combination of both molecules was the only treatment that achieved 100% efficacy against all nematode genera (Cooperia, Ostertagia, Haemonchus and Oesophagostomum spp.). Conclusions: Based on the described PK and pharmacodynamic (PD) assessments, the IVM + LEV combination appears to be a promising pharmacological option for controlling resistant gastrointestinal nematodes in cattle, with the additional potential to delay the progression of nematode anthelmintic resistance. Overall, the study provides original and robust pharmacokinetic and efficacy data that contribute to the optimization of parasite control strategies in cattle. This drug combination strategy may enhance treatment efficacy and contribute to improved parasite control in cattle production systems.

Pharmaceutics doi: 10.3390/pharmaceutics18050629

Authors: An Le Guru R. Valicherla Junmei Zhang Lin Wang Mark K. Donnelly Robert Bies Lisa C. Rohan

Background/Objectives: With the ongoing efforts in supporting the discovery of novel targeted drug delivery systems for the upper region of the female reproductive tract (FRT), it is imperative to understand the local drug disposition pathways. We aim to obtain a comprehensive profile of the drug transporters and drug-metabolizing enzymes in the human ectocervix, uterus, and fallopian tubes, as these factors may substantially influence mucosal penetration, tissue exposure, drug disposition, and the risk of drug–drug interactions. Methods: Gene expression of 12 drug transporters and 21 drug-metabolizing enzymes was quantified using RT-qPCR. Protein expression of highly expressed transporters was assessed using immunohistochemistry (IHC). Results: Among the 12 transporters analyzed, the efflux transporters P-gp, BCRP, and MRP4 exhibited the highest expression across the ectocervix, endometrium, myometrium, and fallopian tubes, with P-gp consistently showing the greatest abundance in all evaluated FRT tissues. Expression of these transporters was significantly higher (6–17×) in myometrium compared with ectocervix. IHC demonstrated strong localization of P-gp, BCRP, and MRP4 to epithelial layers facing the lumen, as well as to stromal and vascular endothelial cells. For drug-metabolizing enzymes, all 21 phase I and II enzymes were detectable across the FRT, and 15 were expressed at comparatively higher levels across all tissue types. These included CYP1A1, CYP1B1, CYP2B6, CYP2C8, CYP2C19, CYP3A4, UGT1A1, UGT1A3, UGT1A4, UGT1A7, UGT1A8, UGT1A10, UGT2B4, UGT2B15, and UGT2B17. Conclusions: The gene expression and localization data obtained from this work may improve our understanding of drug disposition in the FRT, which will inform selection, design, and optimization of drugs intended for targeted delivery within the FRT.

Pharmaceutics doi: 10.3390/pharmaceutics18050627

Authors: Siu-Yin Cheung Aldana Galiyeva Yerkeblan Tazhbayev Tolkyn Zhumagaliyeva Yuliia Bardadym Vladimir Aseyev

Background/Objectives: Tuberculosis treatment remains challenging due to the limited stability and side effects of conventional rifampicin formulations. This study aimed to synthesize and optimize mucoadhesive chitosan–gellan gum nanoparticles for improved rifampicin delivery. The novelty of this work was the introduction of ethanol into the synthesis process, which improved the solubility of rifampicin and contributed to the formation of nanoparticles with the desired physicochemical characteristics. Methods: Rifampicin-loaded chitosan–gellan gum nanoparticles were produced using the polyelectrolyte complex coacervation method. The polymer ratios, drug-to-polymer ratio, temperature and ethanol volume were the main factors that were optimized using the Taguchi method. The physicochemical properties, such as TGA, DSC and FTIR spectroscopy, were investigated. In addition, drug release, mucoadhesive properties and mycobacterial activity against the H37Rv strain of Mycobacterium tuberculosis were examined. Results: Optimization using the Taguchi method produced nanoparticles with a narrow particle distribution (PDI: 0.212 ± 0.021), a satisfactory average size (153 ± 3 nm) and stability against aggregation (zeta potential: 22.94 ± 1.30 mV). A study of the degree of rifampicin release from nanoparticles showed that the drug release is influenced by pH and has a prolonged effect. Drug-loaded nanoparticles exhibited increased mucoadhesion compared with the pure drug. The minimum inhibitory concentration of rifampicin in chitosan–gellan gum nanoparticles for the suppression of the H37RV strain of Mycobacterium tuberculosis was determined. Spectroscopic and thermal analyses confirmed the incorporation of rifampicin in the polymer matrix. Conclusions: The developed chitosan–gellan gum nanoparticles represent a promising mucoadhesive delivery system for rifampicin. The incorporation of ethanol and the use of Taguchi optimization provide an effective strategy for controlling nanoparticle properties and improving drug delivery performance.

Pharmaceutics doi: 10.3390/pharmaceutics18050628

Authors: Ana Alarcia-Lacalle Miguel Ángel Morán-Rodríguez Laura Morata Arantxa Isla Andrés Canut-Blasco Alicia Rodríguez-Gascón

Background/Objectives: Oritavancin therapy for complex infections remains challenging due to the lack of well-established dosing regimens. The objective of this work was to apply PK/PD modeling and Monte Carlo simulation considering different PK/PD targets to identify multiple-dosing regimens that may ensure effective concentrations of oritavancin for the treatment of long-term infections. Methods: Plasma concentration–time profiles were simulated for different regimens (single dose of 1200 mg, 1200 mg followed by 800 mg every 7 days (q7d), 1200 mg followed by 800 mg q10d, 1200 mg q7d, 1200 mg q10d, 1200 mg q14d, 1200 mg q21d, and 1200 mg followed by 1200 mg on day 8, then 1200 mg q14d), and the probability of target attainment (PTA), indicative of treatment success, was estimated. Results: All dosing regimens provided probabilities of target attainment of 100% up to MICs of 0.5 mg/L when AUC0–24/MIC and Cmax/MIC were applied. Considering AUC0–72/MIC, the regimens would be adequate up to an MIC of 0.125 mg/L. For fCmin > MIC, all except 1200 mg q21d were adequate for an MIC of 0.125 mg/L, and 1200 mg day 1 + 800 mg q7d and 1200 mg q10d may be useful to treat infections due to bacteria with an MIC of 0.25 mg/L. Conclusions: More studies involving patients with complex infections are needed to better stablish the relationships among plasma concentrations, MIC values, and clinical outcomes. fCmin > MIC should be investigated as a potential PK/PD target for the treatment of these infections with oritavancin.

Pharmaceutics doi: 10.3390/pharmaceutics18050626

Authors: Esmat Sajjadi Ehsan Sharif-Paghaleh Mohammad Akrami Koorosh Shahpasand Ismaeil Haririan Samane Maghsoudian

Background: Traumatic brain injury (TBI) affects millions of individuals annually and remains a major global cause of neurological disability and death. Tau protein hyperphosphorylation, particularly in its cis conformation, is a major pathological hallmark contributing to neurodegeneration following TBI. Single-chain variable fragments (scFvs), despite their diagnostic potential, suffer from rapid renal clearance and short circulation half-lives, which limit their in vivo performance. PEGylation is therefore employed to prolong systemic circulation and improve the pharmacokinetic behavior of scFvs, enabling more effective brain retention and target engagement. Methods: In this study, we utilized a previously validated anti-cis p-tau scFv antibody fragment, radiolabeled with technetium-99m tricarbonyl (99mTc(CO)3), as a diagnostic tracer to detect tau pathology in TBI rat models. The antibody was conjugated with polyethylene glycol (PEG, 20 kDa); PEGylation efficiency was determined by quantifying the products on SDS-PAGE, and the products were subsequently radiolabeled. Results: Radiochemical purity (RCP) was ~95.4% for the non-PEGylated tracer (99mTc-AININ20) and ~92.7% for the PEGylated form (99mTc-AININ20-PEG), with both showing >90% radiochemical purity consistently. Upon systemic administration, PEGylated scFv was able to cross the blood–brain barrier (BBB) and selectively accumulated in injured regions, as confirmed by single-photon emission computed tomography (SPECT) imaging. Both PEGylated and non-PEGylated scFv tracers showed significantly higher brain uptake in TBI rats compared to healthy controls (p < 0.0001). At 24 h, the PEGylated form exhibited a significantly higher brain signal than the non-PEGylated version (p < 0.0001), indicating improved tracer retention. Biodistribution analysis at 2 h post-injection showed significantly reduced renal clearance for the PEGylated tracer and increased hepatic uptake compared to the non-PEGylated form. At 24 h, in vivo imaging confirmed sustained brain retention, highlighting improved pharmacokinetics and imaging potential. Conclusions: These results support PEGylated scFv as a promising SPECT imaging agent for early detection of tauopathy in TBI, offering enhanced brain retention and improved pharmacokinetics.

Pharmaceutics doi: 10.3390/pharmaceutics18050625

Authors: Diana-Maria Mateescu Dragos-Mihai Gavrilescu Florin Eugen Constantinescu Cristian Oancea Adrian-Cosmin Ilie Roxana Folescu Mihaela-Diana Popa Stela Iurciuc Camelia-Oana Muresan Alexandra Enache

Background/Objectives: BPC-157 (body protection compound 157) is a synthetic pentadecapeptide derived from a gastric protein fragment with reported cytoprotective and regenerative properties across multiple organ systems. Despite over three decades of preclinical research demonstrating consistent biological activity, its pharmaceutical development remains rudimentary, with no approved formulation, no validated dosing regimen, and no completed Phase II clinical trial. This review critically evaluates BPC-157 from a biopharmaceutical and drug development perspective, examining its physicochemical and pharmacokinetic properties, formulation challenges across routes of administration, the pharmacokinetic–pharmacodynamic disconnect that characterizes its preclinical profile, and the regulatory and translational barriers that currently preclude clinical advancement. Methods: A narrative review of the literature was conducted using PubMed/MEDLINE, Embase, and Cochrane Library from database inception to April 2026. Search terms included “BPC-157”, “BPC157”, “body protection compound 157”, “pentadecapeptide”, and “GEPPPGKPADDAGLV”, each combined with “pharmacokinetics”, “formulation”, “biopharmaceutics”, “drug delivery”, “clinical trial”, “toxicology”, and “regulatory”. Patent databases (Espacenet, Google Patents) and regulatory agency websites (FDA, EMA, WADA) were searched independently. Searches were supplemented by forward and backward citation tracking of key references. Articles were selected based on relevance to biopharmaceutical characterization, pharmacokinetics, formulation science, clinical evidence, and regulatory status; pharmacodynamic studies were included insofar as they inform translational development. Evidence was synthesized with emphasis on pharmaceutical characterization, formulation science, and translational feasibility; no formal quality assessment instrument was applied, consistent with the narrative review design. Results: BPC-157 exhibits unusual stability in gastric juice and demonstrates activity via oral, parenteral, and topical routes, yet its human pharmacokinetic profile remains critically undercharacterized despite a recently published formal preclinical ADME study in two species confirming a sub-30-min plasma half-life, linear dose-proportional kinetics, and intramuscular bioavailability of 14–51% depending on species. A plasma half-life of under 30 min—confirmed preclinically and in a preliminary two-subject human pilot—contrasts with prolonged biological effects lasting hours to days—a disconnect with significant implications for dosing strategy and formulation design. No pharmaceutical-grade formulation has been developed or validated. The peptide lacks bcs classification data, permeability characterization, and formal excipient compatibility studies. Available clinical data derive from fewer than 30 subjects across three uncontrolled pilot studies, none of which employed standardized pharmaceutical preparations. Conclusions: BPC-157 presents a compelling but pharmaceutically underdeveloped profile. The primary barrier to clinical translation is not the absence of biological activity, but the absence of fundamental pharmaceutical science: characterized formulations, validated pharmacokinetics, and a coherent drug development strategy. Addressing these biopharmaceutical gaps is a prerequisite for any meaningful clinical program.

Pharmaceutics doi: 10.3390/pharmaceutics18050624

Authors: Svend Buus Eva Greibe Lara Aygen Øzbay Elke Hoffmann-Lücke Niels Henrik Buus

Background/Objectives: Mycophenolic acid (MPA) monitoring may improve organ transplant outcomes, yet clinical implementation is hindered by the complex pharmacokinetics of MPA and a lack of clarity regarding the influence of specific patient factors on drug exposure. While the area under the curve (AUC) is the gold standard for MPA monitoring, it is not easily validated or implemented in routine practice. This pilot project aimed to identify key clinical and biochemical covariates driving pharmacokinetic variability in a renal transplant population. Methods: This prospective study analyzed 103 samples from 66 kidney transplant recipients. To estimate total drug exposure (AUC), a limited sampling strategy was used with plasma samples collected at trough, and then 30 and 120 min post-dose. We performed linear univariate and multivariate regressions to evaluate the impact of patient characteristics (age, sex, body mass index (BMI)) and biochemical measurements (P-albumin, P-creatinine, estimated glomerular filtration rate (eGFR), B-tacrolimus) on MPA-AUC, peak concentrations (Cmax) and trough levels. Results: At 750 mg twice daily, the median MPA-AUC was 43.5 mg·h/L (IQR: 34.5–53.5). After adjusting for dose, P-albumin and age were independent predictors of AUC: P-albumin levels were positively associated with AUC (β = 1.849, p < 0.001), while age showed a modest negative association (β = −0.282). BMI was significantly and inversely associated with trough concentrations (β = −0.137, p = 0.011), indicating that higher BMI is linked to lower trough concentrations. Male sex was associated with significantly lower AUC and Cmax compared to females. Notably, eGFR and B-tacrolimus levels did not significantly influence MPA exposure in this cohort. Conclusions: The covariates BMI, sex, age, and P-albumin significantly influence MPA-AUC. LSS-based AUC monitoring, using 30–60 mg·h/L as a target and with consideration of a few patient-specific factors, could be a pragmatic and feasible approach to improve MMF dosing strategies in kidney transplant recipients.

Pharmaceutics doi: 10.3390/pharmaceutics18050623

Authors: Heng Li Zhiyue Zhang Yu Wang Ting Mou Jiaqi Tian Chong Huang Lu Zhao Zeyang Ge Dandan Wang Chenlu Li Jihong Wang Yanzhen Zheng Lei Tian Chunlin Zong

In the original publication [...]

Pharmaceutics doi: 10.3390/pharmaceutics18050622

Authors: Septika Prismasari Hyeong Jae Kim Jeong Hee Hong Jung Yun Kang

Background/Objectives: Allomyrinasin is a cationic antimicrobial peptide derived from Allomyrina dichotoma larvae with known antibacterial and anti-inflammatory properties; however, its effects on migration-related mechanisms in oral squamous cell carcinoma (OSCC) remain poorly understood. This study investigated the anti-migratory potential of allomyrinasin in OSCC cells, focusing on Na+/HCO3− cotransporter (NBC) activity as a key migratory module. Methods: NBC activity was assessed in YD-38 OSCC cells treated with allomyrinasin. Cell migration was evaluated by wound healing and Transwell assays, and MMP expression. Intracellular reactive oxygen species (ROS), apoptosis-related markers, and lamin A/C expression were analyzed using fluorescence-based assays and gene expression analysis. Results: Allomyrinasin inhibited NBC activity and suppressed cell migration without substantial loss of cell viability. MMP-13 was selectively downregulated among the tested MMPs. Lamin A/C expression was markedly upregulated, suggesting enhanced nuclear stiffness that may restrict confined cell migration. Intracellular ROS levels were elevated, and apoptotic progression was confirmed by increased Annexin V/PI positivity along with downregulation of B-cell lymphoma 2 (BCL2) and upregulation of BCL-2–associated X genes (BAX), through a p53-independent pathway consistent with the TP53-deleted status of YD-38 cells. Conclusions: Allomyrinasin suppresses OSCC cell migration by targeting NBC activity as a key component of the migratory machinery, accompanied by oxidative stress induction and pro-apoptotic signaling. These findings identify allomyrinasin as a potential anti-migratory therapeutic candidate and highlight NBC activity as a promising target for attenuating cancer metastasis.

Pharmaceutics doi: 10.3390/pharmaceutics18050621

Authors: Weiqi Wang Zijiao Yan Yajie Yu Mengjiao Zhou Hejian Xiong Tingting Liu

Metal–organic frameworks (MOFs) possess unique structural tunability, abundant coordination sites, and outstanding biosafety, rendering them highly advantageous for the development of high-performance magnetic resonance imaging (MRI) contrast agents. In light of the significant advancements in MOF-derived theranostic platforms, a comprehensive overview focusing on their classification and clinically oriented applications is urgently required. This review provides an in-depth examination of various categories of MOF-derived contrast agents, including T1, T2, dual-mode, ratiometric and 19F imaging systems, and analyzes the correlation between structural characteristics and imaging performance. Furthermore, it highlights typical MRI-guided therapeutic applications, such as those related to atherosclerosis, bacterial infections, and cancer immunotherapy. The review systematically addresses existing challenges, including issues related to biodegradability, metabolic behavior, and biosafety. It also summarizes the rational design principles for novel MOF contrast agents, aiming to facilitate their transition from fundamental research to clinical applications.

Pharmaceutics doi: 10.3390/pharmaceutics18050620

Authors: Maria-Alexandra Paceana Liliana Mititelu Tartau Ianis Kevyn Stefan Boboc Carmen Nicoleta Oancea Anca Berbecaru-Iovan Cezar Ilie Foia Cosmin Gabriel Tartau Maria Bogdan

Both diabetes mellitus (DM) and stroke are major global health challenges with high morbidity and mortality. DM is a major risk factor for stroke, contributing to both increased incidence and worse clinical outcomes. Incretin-based therapies, including glucagon-like peptide-1 receptor agonists (GLP-1 RAs), as well as dual agonists like tirzepatide, have demonstrated significant cardiovascular benefits, raising interest in their potential cerebrovascular effects. This narrative review examines the pathophysiological links between DM and stroke and summarizes recent clinical evidence on the efficacy of GLP-1 RAs and dual GLP-1/GIP receptor agonists (GLP-1/GIP RAs) in stroke prevention and management. Current evidence from large cardiovascular outcome trials supports the role of GLP-1 RAs in reducing major adverse cardiovascular events, including stroke, primarily in the context of primary and secondary prevention. Findings suggest that semaglutide and liraglutide may reduce non-fatal stroke incidence, decrease hospitalizations, and improve neurological outcomes in patients with prior stroke. Comparative analyses of major trials suggest that, although stroke reduction may be a class effect of GLP-1 RAs, meaningful differences exist between individual agents, likely due to variations in pharmacokinetics, receptor affinity, and study populations. Additionally, much of the evidence in acute stroke derives from early-phase or ongoing trials, warranting cautious interpretation. Novel therapies, including orforglipron and retatrutide, as well as combinations like Maridebart cafraglutide and CagriSema, may expand future therapeutic options for individuals at high cerebrovascular risk. GLP-1-based therapies show promising neurovascular effects, but large-scale, long-term studies are needed to define their role in post-stroke management and cerebrovascular risk reduction. Overall, GLP-1 RAs should currently be regarded primarily as agents for long-term vascular risk reduction rather than established therapies for acute stroke. While potential neuroprotective effects are emerging, these require confirmation in adequately powered randomized trials. Future studies should aim to identify the patient subgroups most likely to benefit and to determine whether specific agents confer advantages in acute cerebrovascular contexts. A better understanding of the mechanisms underlying potential neuroprotection will be essential to determine whether these therapies can be effectively integrated into stroke management strategies.

Pharmaceutics doi: 10.3390/pharmaceutics18050619

Authors: Margarita C. Dinamarca Boris Sevarika Scott McNeil

Background/Objectives: The blood–brain barrier (BBB) presents a major challenge for delivering therapeutics to the central nervous system (CNS) due to its highly selective permeability. Human brain microvascular endothelial cells (hBMECs), the principal cellular component of the BBB, tightly regulate molecular transport and restrict the entry of many CNS-targeted therapies. Lipid-based nanoparticles have emerged as promising carriers for BBB transport because of their biocompatibility, tunable surface properties, and cargo encapsulation capabilities. One strategy to enhance nanoparticle transport involves surface functionalization with ligands that exploit endogenous transcytosis pathways. Mannose-6-phosphate (M6P), a glycan implicated in the brain entry of certain proteins and viruses, represents a potential targeting ligand for this purpose. Methods: In this study, we established a physiologically relevant in vitro BBB model using human-induced pluripotent stem cell-derived brain microvascular endothelial cells (hiPSC-BMECs) to evaluate M6P-functionalized liposomes for BBB transport. Fluorophore-labeled liposomes were used to monitor nanoparticle uptake and transcytosis. Results: M6P-functionalized liposomes exhibited significantly enhanced uptake in hiPSC-BMECs compared with non-functionalized control liposomes. Pharmacological inhibition studies supported the involvement of a clathrin-sensitive endocytic pathway. Transcytosis assays demonstrated enhanced BBB crossing of M6P-functionalized liposomes, with transport increasing according to ligand density and reaching approximately 55% of the transport observed for transferrin under the same experimental conditions. Following transcytosis, intact M6P-functionalized liposomes showed significantly higher uptake by downstream hiPSC-derived neurons and astrocytoma cells compared with control formulations. Conclusions: Together, these findings support M6P-functionalization as a promising strategy to enhance liposome uptake and transcytosis across a human-relevant in vitro BBB model. This work provides a proof-of-concept framework for the development and optimization of glycan-functionalized nanocarriers for CNS-directed delivery.

Pharmaceutics doi: 10.3390/pharmaceutics18050618

Authors: Søren Aleksander Friederici Dahl Elke Hoffmann-Lücke Birgitte Klug Albertsen Eva Greibe

Background: PEG-Asparaginase and Erwinia asparaginase are enzyme-based anticancer therapies used in the treatment of acute lymphoblastic leukaemia (ALL) and lymphoblastic lymphoma (LBL), where adequate plasma enzyme activity is required for therapeutic efficacy. In many study groups, therapeutic drug monitoring is routinely applied due to pharmacokinetic variability and the risk of hypersensitivity reactions followed by increased clearance and insufficient treatment. In clinical practice, samples may be exposed to prolonged transport and variable pre-analytical conditions. Knowledge on pre-analytical stability is important for correct interpretation of PEG-Asparaginase and Erwinia asparaginase activity in plasma. This study aimed to evaluate the in vitro stability of PEG-Asparaginase and Erwinia asparaginase under pre-analytical conditions. Methods: Three experimental stability studies were conducted at two activity levels. Enzyme stability in plasma was assessed during storage at 4 °C and 20 °C for up to 14 days and following three freeze–thaw cycles. Stability in whole blood prior to centrifugation was evaluated over 24 h. Enzyme activity was measured using a validated spectrophotometric assay, and stability was defined as a deviation within ±15% of baseline activity. Results: Both enzymes remained stable in plasma for up to 14 days at 4 °C and 20 °C, and no clinically relevant reduction in enzyme activity of freeze–thaw cycling was observed. In whole blood, Erwinia asparaginase and high-activity PEG-Asparaginase remained stable for 24 h at 20 °C, whereas low-activity PEG-Asparaginase showed a reduction in activity of approximately 22%, mainly within the first two hours. Conclusions: PEG-Asparaginase and Erwinia asparaginase are stable in plasma for up to 14 days at room temperature, enabling shipment of plasma samples by mail. However, prompt centrifugation is recommended for samples with low PEG-Asparaginase activity to ensure accurate therapeutic drug monitoring.

Pharmaceutics doi: 10.3390/pharmaceutics18050617

Authors: Chenxiao Chu Xingting Fan Xiaoman Zhang Tongyao Zhao Yuying Wang Xing Tang Yu Zhang Tian Yin

Background: Diabetic foot ulcers (DFUs) are difficult to heal because hyperglycemia-associated pathological exudation, excessive oxidative stress, chronic inflammation, and impaired cellular regeneration jointly maintain a nonhealing wound microenvironment. This study aimed to develop and evaluate a composite hydrogel containing Periplaneta americana (PA) extract and calamine as a Zn2+ source for dynamic modulation of the diabetic wound microenvironment and promotion of tissue repair. Methods: A PA composite hydrogel was prepared and assessed in vitro for reactive oxygen species (ROS)-scavenging activity and effects on fibroblast migration. Therapeutic efficacy was further evaluated in a streptozotocin (STZ)-induced diabetic full-thickness wound model in rats. Wound closure, histological remodeling, oxidative stress markers, inflammatory mediators, growth factors, angiogenesis, and AGEs-RAGE/NF-κB pathway-related changes were analyzed. Results: The composite hydrogel reduced excessive intracellular ROS and enhanced fibroblast migration in vitro compared with pathological-condition controls. In diabetic rats, topical treatment accelerated macroscopic wound closure and promoted more mature histological repair. Mechanistic analyses showed attenuation of the AGEs-RAGE/NF-κB signaling axis, accompanied by restoration of superoxide dismutase activity, reduction of malondialdehyde levels, and suppression of TNF-α-associated inflammatory responses. The improved wound microenvironment was associated with increased expression of platelet-derived growth factor and basic fibroblast growth factor, enhanced cellular proliferation, and increased neovascularization within the wound tissue. Conclusions: The PA composite hydrogel improved diabetic wound healing by concurrently alleviating oxidative and inflammatory barriers and enhancing regenerative signaling. These findings suggest that microenvironment-modulating PA composite hydrogel systems may represent a promising therapeutic strategy for refractory diabetic wounds.

Pharmaceutics doi: 10.3390/pharmaceutics18050616

Authors: Meitong Lin Wenlu Yu Ke Zhang Jiayi Wu Xingtong Chen Yuke Pan Yujie Tian Liangjia Zeng Haorui Yuan Xiaofei Hu Xuhua Tan

Cataract remains the preeminent cause of reversible blindness globally, with cataract extraction and intraocular lens (IOL) implantation serving as the definitive surgical intervention. Nevertheless, its long-term efficacy is undermined by formidable postoperative complications, specifically posterior capsule opacification (PCO) and endophthalmitis, which necessitate effective prophylactic strategies. IOL modification has emerged as a pivotal paradigm to effectively mitigate these complications. Current approaches encompass surface modification, drug delivery IOLs, and photo-responsive IOLs. Driven by the rapid interdisciplinary convergence of materials science, ophthalmology and pharmacology, the field has also evolved to have combined modification strategies and multifunctional systems. This review provides a comprehensive overview of the recent progress in IOL modification for postoperative complication prophylaxis. By categorizing recent advancements into three major types—surface modification, drug delivery systems, and photo-responsive IOLs—we critically evaluate their mechanisms, advantages, and limitations. Furthermore, we offer strategic insights to accelerate the development of IOL modification and bridge the gap between innovation and clinical translation.

Pharmaceutics doi: 10.3390/pharmaceutics18050615

Authors: Eliza Grațiela Popa Liliana Mititelu Tartau Alina Diana Panainte Larisa Păduraru Andreea Crețeanu

Four decades have elapsed since orally disintegrating tablets (ODTs) were first formulated as the emulsion/type Lyoc tablet, a porous mass intended to rapidly disperse in saliva. Following the lyophilization process, new formulations of ODTs were designed, intending to make a simpler and more reproducible formulationZydis, LBL-Flash, Quicksolv, and, more recently, Zydis Ultra. Lyophilization is widely recognized as an effective technique for the development of ODTs, due to its ability to produce highly porous structures that enable rapid disintegration and improved patient compliance. However, its advantages should be considered in relation to other manufacturing methods, as each technology presents specific trade-offs in terms of cost, scalability, mechanical strength, drug loading capacity, and process robustness. In line with the modern sustainable and green pharmacy trend, new raw materials have gained attention as excipients for lyophilized ODTs; these materials include certain plant derivatives, but also performant excipients with newly discovered functionalities. At present, a new generation of ODTs is available in the form of Self-Nanoemulsifying Lyophilized Tablets (SNELTs), which bring the advantages of Self-Nanoemulsifying Drug Delivery Systems (SNEDDS) into ODTs via the lyophilization method. The technique is mostly applicable to low-solubility drugs formulated as nanoemulsions, which are absorbed onto solid carriers and further lyophilized, forming the final ODT. Despite its limitations (expensive, time-consuming, and high product friability), lyophilization is being continuously developed nowadays, in combination with other techniques (3D printing, mucoadhesion, or electrospinning), building hybrid platforms for the modern ODTs of the future.

Pharmaceutics doi: 10.3390/pharmaceutics18050614

Authors: Sarigama Rajesh Gwo Yaw Ho Ravindu Fernando Poh Yi Gan Jessica Wu Jiali Zhai Joshua D. Ooi Calum J. Drummond Nhiem Tran

Background: Platinum-based chemotherapy, including cisplatin and carboplatin, is widely used to treat various cancers, including ovarian cancer. However, its clinical application is limited by dose-limiting toxicities and resistance, with a poor 5-year overall survival rate for ovarian cancer (35–40%). In this study, we used ionisable lipids and developed pH-responsive lipid nanoparticles (LNPs) to address platinum-resistance in ovarian carcinoma. Methods: Cisplatin was loaded into three LNP systems containing monoolein (MO) and synthetic cationic ionisable lipids (OE-Mo, OA-Py, and OA-Pi) dispersed in Pluronic F-127 with 0.9% NaCl. Cisplatin-loaded LNPs (Cis-OE-Mo-NP, Cis-OA-Py-NP, and Cis-OA-Pi-NP) were characterised for size, zeta potential, and internal mesophase structure. Encapsulation efficiencies were determined via HPLC after removing free drug by ultrafiltration. In vivo efficacy was tested using cisplatin-resistant human patient-derived xenograft (PDX) models. Results: The LNPs were well dispersed with particle size of 219–250 nm and a drug loading of ~1.2 mg/mL. Encapsulation efficiencies were 62%, 59%, and 64%, for Cis-OE-Mo-NP, Cis-OA-Py-NP, and Cis-OA-Pi-NP, respectively. Small angle X-ray scattering (SAXS) results showed that the LNPs are pH responsive with structural transitions from a cubic to a hexagonal phase at an acidic pH. Among the tested formulations, Cis-OA-Py-NP resulted in the most significant reduction in tumour volume by ~60% compared to treatment with cisplatin alone. However, they also showed significant toxicity, including >10% weight loss and gross lung and kidney damage, as confirmed by histology. Conclusions: These findings highlight the potential of Cis-OA-Py-NP in reducing tumour volume but underscore the need for further optimisation to improve safety and therapeutic applicability.

Pharmaceutics doi: 10.3390/pharmaceutics18050613

Authors: Qizheng Wang Pengcheng Guo Tianci Hu Longjie Li Tingxi Zhu Yue Pan Xiaoqiang Xiang Jianxin Wang

Background: Dual-release sustained formulations enable rapid drug release for prompt therapeutic onset while retaining the characteristics of sustained-release dosage forms. However, due to the complexity of this dosage form, conventional trial-and-error approaches fail to mitigate development risks or improve the success rate of bioequivalence studies of the generic product. Accordingly, the present study aims to investigate the feasibility of guiding a dual-release generic formulation screening through establishing a quality-related media condition through construction of PBPK models and IVIVR for the reference product. Methods: Here, Difene® was selected as the reference product, and GastroPlusTM was employed as the simulation platform. Pharmacokinetic data obtained from the literature and in vitro dissolution test results were integrated to construct the PBPK model for the reference product and establish IVIVR in different media, respectively. A quality-related media condition was determined for formulation screening of the generic product. A pharmacokinetic study in beagle dogs was then conducted to evaluate the bioequivalence between the generic and the reference product. Results: In the PBPK modeling and IVIVR study, the PBPK model was successfully established. The IVIVR for the pH 4.0–pH 6.0–pH 6.8 media was optimal in all media conditions, with fold error ratios of 1.11, 0.86, and 1.11 for Cmax, AUC, and Tmax, respectively, all falling within the 0.80–1.25 range. Employing this medium as the quality-related media, the optimized generic product exhibited an f2 factor of 76 with the reference product in vitro. Pharmacokinetic studies in beagle dogs demonstrated that the geometric mean ratios and 90% confidence intervals for AUC and Cmax of the generic product versus the reference product were within the 80.0–125.0% range. No statistically significant difference was observed for Tmax, indicating bioequivalence between the two products. Conclusions: Overall, our study provides a strategic approach for generic development and a novel research framework for the generic development of other dual-release formulations.

Pharmaceutics doi: 10.3390/pharmaceutics18050612

Authors: Mashan Almutairi Ahmed Adel Ali Youssef Gehad M. Subaiea Ahmed Alobaida Sultan Almuntashiri

Background/Objective: The introduction of Ketoconazole (KZ, Nizoral®) in 1977 by Janssen Pharmaceutica marked a significant milestone in medical mycology as the first broad-spectrum oral antifungal agent. However, KZ is a highly lipophilic compound, presenting significant challenges in the development of efficient topical formulations. Moreover, oral KZ has undergone labeling revisions and market withdrawal due to serious hepatic side effects. This study aimed to design, optimize, and evaluate KZ-loaded nanoemulsions (NEs; KZ-NEs) as a delivery platform that could improve skin bioavailability and antifungal activity. Methods: Optimized KZ-NEs were converted to a mucoadhesive formulation (KZ-NEC) by the addition of Carbopol® 940 NF to enhance the adherence of the formulations to the skin surface. NEs were evaluated concerning physical appearance, globule size, polydispersity index, zeta potential, pH, viscosity, and drug content. Optimized KZ-NE and lead KZ-NEC formulations were further evaluated for in vitro release, ex vivo skin permeation and deposition, skin irritation, and in vivo studies. Results: In vitro release studies revealed that nanocarrier systems provided a sustained release of KZ over 24 h. The ex vivo permeability coefficients of KZ from the optimized KZ-NE and lead KZ-NEC formulations were approximately four- and three-fold greater than that achieved with the marketed cream formulation, respectively. In addition, the Cmax of the lead KZ-NEC formulation (14.4 ± 1.1 μg/mL) was significantly higher (p < 0.05) compared with the marketed cream formulation (10.5 ± 0.5 μg/mL). Moreover, in vitro antifungal susceptibility testing showed that KZ demonstrated improved antifungal efficacy when incorporated into the KZ-NE and KZ-NEC formulations. Neither of the NE-based formulations caused any alterations in skin color or morphology during the 24 h visual observation period. Both NE-based formulations were stable for 90 days (the last time-point tested) at three different storage conditions. Conclusions: NE-based formulation could serve as an effective topical delivery platform for KZ and could improve therapeutic outcomes for patients with topical fungal infections.

Pharmaceutics doi: 10.3390/pharmaceutics18050611

Authors: Shery Jacob Hiral Shah Anroop B. Nair

The low water solubility of numerous drug candidates and phytochemicals continues to pose a significant challenge in pharmaceutical development, greatly limiting their bioavailability and therapeutic performance. This review presents a detailed overview of formulation strategies aimed at improving the solubility and dissolution of poorly aqueous-soluble compounds. The biopharmaceutics classification system and the relevance of in vitro–in vivo correlation, as well as key challenges in formulation development, are briefed. Solid-state and particle engineering approaches, including micronization, supercritical fluid technology, electrospinning, and cryogenic techniques, are discussed. Extensive critical examination of amorphous solid dispersions and their preparation methods, as well as crystallization inhibition strategies, is covered. Cocrystallization is highlighted as a promising approach, with emphasis on design principles and preparation methods. Various solubilization techniques, such as pH modification, cosolvency, hydrotropy, micellar solubilization, and cyclodextrin-based complexation, including advanced hybrid systems, are also explored. Emerging solvent platforms, such as deep eutectic systems and lipid-based and nanotechnology-driven approaches, are reviewed for their role in improving solubility and drug delivery. Additionally, enabling technologies such as liquisolid systems and hydrophilic polymers are addressed. Despite notable progress, limitations such as scalability, reproducibility, regulatory constraints, and long-term safety persist. Overall, this review provides integrated insights into formulation design approaches to enhance the solubility and therapeutic efficacy of poorly soluble drugs.

Pharmaceutics doi: 10.3390/pharmaceutics18050610

Authors: Hilal Gülsena Nur Akkus Ayse Betül Bingol Büsra Oktay Buse Ozsan Ahmet Akif Kızılkurtlu Azime Erarslan Fatih Ciftci Cem Bülent Ustündag

Background/Objectives: Guided bone regeneration (GBR) in dental applications requires scaffolds that possess balanced mechanical strength, controlled biodegradability, and excellent biological performance; therefore, this study aims to develop and evaluate a multilayered biofunctional dental membrane designed to enhance mechanical, biological, and antibacterial performance. Methods: The multilayered membrane was fabricated using sequential electrospinning and electrospraying techniques to form a polycaprolactone (PCL)/Collagen first layer and a polyvinyl alcohol (PVA)/Collagen/Hydroxyapatite (HAp) second layer, topped with a final electrospray coating of PVA/Amoxicillin. Characterization was performed via SEM, FTIR, and EDS, followed by evaluations of tensile properties, swelling behavior, hydrolytic degradation, in vitro drug release, disk diffusion antibacterial activity against Staphylococcus aureus and Escherichia coli, and 7-day L929 fibroblast cytocompatibility (ANOVA/Tukey, p < 0.05). Results: SEM, FTIR, and EDS analyses confirmed uniform nanofiber morphology, homogeneous HAp distribution, and successful integration of bioactive compounds. The membrane exhibited a maximum tensile strength of 15.17 N, strain of 25.24%, and stress of 2.16 MPa, while swelling reached ~100% within 2 h and degradation stabilized around 4% weight loss after 48 h. Drug release profiles showed a rapid amoxicillin release in the first 50 min, plateauing at approximately 4.5 mg/L by 350 min, with distinct antibacterial inhibition zones, and the PCL/Col–PVA/Col/HAp–PVA/Amox group demonstrated the highest cell viability (~140%) after 7 days, significantly exceeding the control groups (p < 0.01). Conclusions: These quantitative findings validate the fabricated multilayered membrane’s potential as a mechanically robust, biodegradable, antibacterial, and bioactive scaffold for advanced guided bone regeneration in dental applications.

Pharmaceutics doi: 10.3390/pharmaceutics18050609

Authors: Mariam Murad Pearl Wasif Laura Dempsey G. Roshan Deen Alexandra E. Butler Stephen L. Atkin

Objective: Sporopollenin exine capsules (SpECs) have been used to encapsulate active pharmaceutical agents for oral drug delivery. This study investigated whether fragrance encapsulated within SpECs prolonged perceived fragrance intensity compared with fragrance oil alone. Methods: A double-blind, placebo-controlled, randomized pilot study was conducted (clinical trial number: NCT07383337); ten healthy female participants (mean age 35.4 ± 5.6 years) received fragrance with SpEC-encapsulated fragrance (SpECs) on one wrist and fragrance alone (control) on the contralateral wrist. The fragrance intensity was assessed using a visual analogue scale (0–10) by both the participants and an independent blinded reviewer at baseline and after 2, 4 and 8 h. Paired Wilcoxon signed-rank tests and linear mixed-effects models were used for analysis. In vitro cytotoxicity was assessed using an ATP viability assay in human bronchial epithelial (BEAS-2B) cells exposed to SpECs or raw Lycopodium clavatum spores. Results: There were no significant differences between the formulations at baseline. From 2 h onward, SpECs was associated with significantly a higher fragrance intensity compared with the control for both participant-rated (p = 0.03 at 2 and 4 h; p = 0.005 at 8 h) and reviewer-rated assessments (p = 0.02 at 2 h; p = 0.01 at 4 h; and p = 0.008 at 8 h). Mixed-model analyses suggested a greater decline in intensity for control at 8 h for reviewer-rated assessments. In vitro, raw spores significantly reduced cell viability (as an indicator of potential allergenicity), whereas SpECs did not differ from control. Conclusions: Fragrance encapsulation within SpECs significantly prolongs measured fragrance intensity with no evidence of cytotoxicity. These findings support the potential of SpECs as a safe and effective sustained-release platform for topical fragrance formulations.

Pharmaceutics doi: 10.3390/pharmaceutics18050608

Authors: Shuai Li Xiaoyan Wang Li Zhi Mohammed Shameem Dingjiang Liu

Background/Objectives: Recombinant adeno-associated virus (AAV) stands at the forefront of gene therapy development, requiring stable formulations to support the expanding therapeutic applications. The growing diversity of serotypes and engineered capsids often creates complex challenges for formulation development, thus demanding innovative formulation development strategies beyond traditional manual approaches to characterize a large formulation design space quickly to discover stable formulations. Methods: Here, we address this critical need through a high-throughput automation platform that dramatically enhances formulation development efficiency and capability through rapid formulation preparation and high-throughput AAV analytics. This system prepares 96 distinct formulations in 40 min and completes AAV compounding in 20 min per plate, with precise control of pH, buffer components, and AAV titers. Results: In a proof-of-concept formulation development study using AAV1, we screened 128 formulations across multiple buffer systems, pH ranges, and excipient combinations. This comprehensive approach successfully identified optimal stable high-titer AAV1 formulations (1.2 × 1014 vector genome (vg)/mL) that maintained stability under frozen, refrigerated, and room temperature storage conditions. Conclusions: Our study demonstrated that this automation platform combined with high-throughput AAV analytics significantly accelerates formulation development, conserves AAV material, and enables systematic exploration of broader formulation design space. It allows us to achieve identification of robust and stable AAV formulations within a timeframe unmatched by traditional formulation development approaches.

Pharmaceutics doi: 10.3390/pharmaceutics18050607

Authors: Hao-Zhe Yu Guan-Yong Deng Nan Gao Li-Hong Fan Jian-Wen Wang Xing-Jian Liu Wei Zhang Shi-Lin Tian Yu-Xiong Weng He-Shuang Dai Yi-Wen Zhang Huan Deng

Background: Photothermal therapy (PTT), a highly efficient and controllable method with minimal drug resistance, transforms near-infrared (NIR) radiation into heat. This process exerts antibacterial effects, aids in tissue repair, and promotes healing. Methods: Our study presented a novel kind of composite wound dressing that incorporated adhesive conductive hydrogel combined with piezoelectric film for NIR-responsive applications. The inherent adhesiveness of the hydrogel ensured robust anchoring of the piezoelectric film to both hydrogel matrix and wound site. Its conductivity enabled synergistic endogenous electrical stimulation with the piezoelectric film, while also serving as therapeutic layer to augment hemostasis, analgesia, and antibacterial activity. Results: The hydrogel’s capacity for moisture retention and exudate absorption sustained optimal wound environment, thereby supporting debridement and recovery. Furthermore, the piezoelectric film possessed excellent photothermal properties and transferred heat to the hydrogel through heat conduction to enhance antibacterial activity and promote wound healing. The in vitro and in vivo experiments confirmed that the composite dressing exhibited strong promotion effect on wound healing under NIR irradiation. Conclusions: In summary, our research provided a new strategy for developing advanced piezoelectric biomaterials with great clinical potential for wound healing.