Ferroptosis is an iron-mediated cell death process driven by lipid peroxidation, yet its antitumor potential is often counteracted by the limited endogenous H2O2 content, the strict catalytic conditions required for the Fenton reaction, and protective autophagy. Notably, sustained autophagy drives ferritin degradation and iron release, thereby amplifying ferroptotic signaling. To exploit this mechanism, we designed a CD44-targeted nanoplatform, Cur@MG@HA, using hyaluronic acid for tumor-specific delivery. The system co-delivers glucose oxidase (GOx) and curcumin (Cur) within a Fe-Cu MOF. Following cellular uptake, GOx catalyzes the oxidation of intratumoral glucose to produce gluconic acid and H2O2. The acidic microenvironment promotes the Fe/Cu-mediated Fenton reaction, converting H2O2 into OH and initiating ferroptosis. The resulting oxidative stress induces autophagy, a process further potentiated by Cur. Enhanced autophagy accelerates ferritin degradation, thereby elevating labile iron levels and establishing a self-reinforcing cycle that amplifies lipid peroxidation, culminating in ferroptotic cell death. In vitro and in vivo experiments demonstrate that the self-reinforcing cycle between autophagy and ferroptosis significantly enhances tumor suppression. This study provides a mechanistic basis for synergistic therapy through redox-metabolic modulation and suggests that targeting iron-autophagy crosstalk with nanomaterials represents a promising strategy for cancer treatment.
PURPOSE:Rebamipide (REBA) is used in the treatment of dry eye disease (DED) and is available as Mucosta® ophthalmic suspension that is widely used in Japan. However, certain challenges such as blurred vision, less permeation due to large particle size, frequent administration because of nasolacrimal drainage, and bitter taste were observed after topical instillation. So, in this study, rebamipide nanosuspension (REBA-NS) bearing in-situ gel (REBA-NS gel) was developed for addressing the challenges as well as ease of application. METHODS:REBA-NS was prepared using the acid-base neutralization method, employing HPMC as a stabilizer and optimized using the quality-by-design approach. Further characterization and evaluation of REBA-NS was carried out, and the optimized REBA-NS was loaded to gel using gellan gum, which has a property of forming an in-situ gel after contact with the simulated tear fluid. Finally, the in-vitro and in-vivo studies of the REBA-NS gel were performed. RESULTS:REBA-NS exhibited nanosized particles (<270 nm) that may help in enhanced permeation, low polydispersity index (<0.25) with high drug content (>80%). Moreover, the REBA-NS gel exhibited mucoadhesiveness, sustained drug release, sterile nature, optimum pH (∼7.2), and osmolality (∼216 mOsm/kg) required for ocular administration, which may aid in the retention of REBA. Finally, REBA-NS gel on the DED-induced rats depicted an increased tear secretion and decreased corneal fluorescein scoring by 1.39-folds and 9.12-folds, respectively, compared to the REBA solution on day 7 of treatment. CONCLUSION:In a nutshell, REBA-NS gel was found to be a promising approach for treating DED.
A physiologically based pharmacokinetic (PBPK) model for carbamazepine (CBZ) was calibrated against an intravenous clinical dataset. Absorption was characterised using the Advanced Dissolution, Absorption and Metabolism (ADAM) model coupled to the diffusion layer model and a particle population balance framework, incorporating formulation-specific particle size distributions. Model verification was performed against 36 clinical datasets spanning single-dose (SD), and multiple-dose (MD) oral administrations across multiple dosage forms. The model was benchmarked against a published CBZ PBPK model (Yin et al., 2024) and further verified through five drug-drug interaction scenarios involving cytochrome P450 modulation. Virtual bioequivalence trials (2 × 2 crossover, n = 24, 100 replicates) were conducted for oral suspensions and immediate-release tablets to compare SD and MD discriminatory power, and to evaluate partial AUC from 0 to 2 h (pAUC0-2h) against conventional bioequivalence metrics. The model matched or outperformed Yin et al. across all scenarios. SD designs consistently demonstrated superior discriminatory power over MD designs (contradicting the proposal that MD studies are more discriminating for autoinducing CBZ); despite autoinduction progressively increasing apparent clearance, formulation-related differences in absorption were attenuated at steady state. pAUC0-2h showed higher sensitivity than Cmax for detecting absorption rate differences. These findings support SD study designs for CBZ bioequivalence assessment and identify pAUC0-2h as a clinically meaningful complementary metric.
Porcine epidemic diarrhea virus (PEDV) can infect pigs of all ages, causing porcine epidemic diarrhea and resulting in significant losses to the global pig farming industry. The virus causes epidemic diarrhea and represents a significant threat to the global swine industry. To enhance vaccine safety and protective efficacy, a PEDV nanoparticle vaccine (PEDV-MS2 VLP) based on MS2 bacteriophages was developed. The MS2 phage coat (MS2 VLP) self-assembles into nanoparticles within Escherichia coli. It is conjugated to the PEDV RBD antigen via the SpyTag-SpyCatcher system, forming a structure mimicking the natural virus. Subcutaneous immunization of BALB/c mice with PEDV-MS2 VLP at days 0 and 14 resulted in a 100-fold increase in neutralizing titers and robust antigen-specific IgG responses. Additionally, PEDV-MS2 VLP induced high levels of cytokines (TNF-α, IFN-γ). The vaccine showed no significant toxicity to the liver, kidneys, or heart in mice. In summary, PEDV-MS2 VLPs demonstrate excellent safety and are a novel candidate vaccine for porcine epidemic diarrhea virus.
Baicalin (BAI) is a natural flavonoid with hepatoprotective potential; however, its poor aqueous solubility and low oral bioavailability limit its therapeutic efficacy. Here, a carrier-free, self-assembled mangiferin (MAN) hydrogel was developed for oral BAI delivery without additional polymeric carriers or chemical crosslinkers. The resulting BAI@MAN HG formed a porous network with favorable hydration and viscoelastic properties while retaining antioxidant activity. FTIR, Raman spectroscopy, XRD, and DSC collectively revealed altered molecular organization, reduced crystalline order, and changes in the solid-state environment of BAI following hydrogel formation, consistent with a possible noncovalent association between BAI and MAN. BAI@MAN HG enhanced the apparent solubility, gastrointestinal release, and oral absorption of BAI, resulting in greater systemic exposure than free BAI. In a preventive mouse model of acetaminophen-induced acute liver injury, oral administration of BAI@MAN HG markedly improved liver morphology, reduced hepatocellular damage, normalized serum transaminase levels, and alleviated oxidative stress and inflammation. These protective effects were accompanied by restoration of redox homeostasis and reduced hepatic levels of HMGB1, NF-κB and other inflammatory mediators. Overall, MAN functioned as both a structural gelator and a bioactive matrix, providing a promising carrier-free platform for enhancing the oral delivery and in vivo efficacy of poorly water-soluble bioactives.
The low oral bioavailability of Dihydromyricetin(DMY), resulting from its instability and poor absorption in the gastrointestinal tract, limits its pharmaceutical applications. In order to enhance the oral bioavailability of DMY, a DMY@CD-CS/TPGS nano-delivery system was successfully prepared using β-cyclodextrin grafted with chitosan (CD-CS) and Tocopheryl Polyethylene Glycol Succinate(TPGS). The DMY@CD-CS/TPGS-NPs have a uniform particle size distribution and good stability. In vitro release studies demonstrated that the NPs exhibited a sustained-release profile under acidic conditions. In vivo and in vitro tests showed that the NPs could promote drug uptake and internalization in Caco-2 cells through multiple endocytic pathways, and disrupt epithelial tight junctions, thereby increasing paracellular absorption. Additionally, it exhibits excellent mucosal adhesion, thereby prolonging drug retention time. The intestinal segment distribution and unidirectional perfusion experiment further demonstrated that the NPs enhanced drug uptake in the duodenum, jejunum, and ileum, and increased both the absorption rate constants and the apparent permeability coefficients. Pharmacokinetic studies demonstrated that DMY@CD-CS/TPGS-NPs significantly enhanced the oral absorption of DMY, increasing the Cmax, AUC0-t, and t1/2 by 7.12-fold, 1.25-fold, and 14.26-fold, respectively. The relative oral bioavailability was 250.39%, representing a 2.50-fold improvement over DMY. Overall, the CD-CS/TPGS nano-delivery system represents a promising strategy for improving the oral delivery of poorly absorbable compounds such as DMY.
Atopic dermatitis (AD) is a chronic, relapsing inflammatory skin disorder characterised by pruritus, epidermal barrier dysfunction, and immune dysregulation, significantly impacting quality of life. Conventional therapies, including corticosteroids, calcineurin inhibitors, and systemic immunosuppressants, provide mainly symptomatic relief with limited long-term efficacy and potential adverse effects. Additionally, Janus kinase (JAK) inhibitors such as Ruxolitinib (RUX) (Opzelura® 1.5 % w/w) have improved treatment outcomes, but their prolonged use causes folliculitis and increased risk of infections. To solve this existing problem, we developed a RUX-NEG to enhance the localised epidermal delivery, along with reducing systemic exposure by employing a low-dose NEG, which has a prolonged release profile and better therapeutic efficacy. Nanoemulsions containing 0.5, 0.75 and 1 % w/w RUX were optimised using a BOX-Behnken design, yielding nanosized droplets (∼16 nm) with a narrow size distribution. Incorporation of 1.5 % w/w high-molecular-weight hyaluronic acid provided suitable rheological characteristics, including pseudoplastic and thixotropic behaviour for improved topical application. The developed RUX-NEG exhibited notable antioxidant potential and significantly attenuated reactive oxygen species generation, mitochondrial membrane depolarisation, and cell death in LPS-stimulated HaCaT cells. Additionally, it suppressed pro-inflammatory mediators (TNF-α, IL-6, IL-8, IL-1β) and modulated NF- κB signalling pathways. Dermatokinetic evaluation demonstrated enhanced drug retention within AD lesions. Furthermore, in a 2,4-DNCB-induced AD mouse model, RUX-NEG effectively reduced inflammatory cytokines (TNF-α, IL-4, IL-6, IL-13) (ELISA), inhibited JAK1/JAK2 signalling (western blot) and improved histopathological alterations. Collectively, these findings highlight the potential of RUX-NEG as an effective topical delivery system for improving therapeutic outcomes and safety in the management of AD.
Tuberculosis (TB) remains a major global health threat, compounded by prolonged treatment regimens, drug toxicity, and the emergence of multidrug-resistant Mycobacterium tuberculosis strains. In this study, we present a novel therapeutic strategy integrating co-crystallization and nanocarrier engineering to enhance the intracellular delivery and antimicrobial efficacy of rifampicin and berberine. Rifampicin-Berberine (RIF-BBR) co-crystals were synthesized via solvent-assisted grinding, and their formation was confirmed through PXRD, NMR, FTIR, DSC, TGA, and TEM analyses, revealing a new crystalline phase stabilized by non-covalent interactions and improved physicochemical properties. The co-crystal was subsequently encapsulated into chitosan-based nanohydrogels and liposomes, yielding nanocarriers with high entrapment efficiencies, favorable particle sizes, and enhanced colloidal stability. In vitro drug release studies demonstrated pH-responsive and sustained release from the nanohydrogel system, while the liposomes exhibited an initial burst release suited for rapid drug availability. Antimicrobial evaluation against M. tuberculosis H37Rv showed that the nanoformulated co-crystal significantly improved inhibitory and bactericidal activity compared to free rifampicin, berberine, and the co-crystal alone, with the liposomal formulation displaying the lowest MIC and MBC values. Collectively, these findings underscore the potential of combining co-crystal engineering with nanocarrier-based delivery to overcome the limitations of conventional TB therapy. This dual strategy enhances intracellular drug accumulation, potentiates antimicrobial activity, and provides a promising foundation for developing more effective, targeted, and patient-compliant anti-tuberculosis treatments.
Quality by Design (QbD) is a systematic approach that builds quality into pharmaceutical products from early development. Within this framework, multivariate data analysis (MVDA), implemented here as a second-order polynomial response-surface model, has long served as the core modeling technique. However, this fixed functional form is limited in capturing complex nonlinear behavior. Artificial neural networks (ANNs) offer a nonlinear alternative. In the small-sample regime typical of DoE-based pharmaceutical development, however, ANNs are prone to overfitting, and generalization to independent test data is not reliably ensured. To address both limitations, the Autoregressive Auxiliary Layer Network (AXLN) and a Hybrid MVDA-AXLN Model are proposed. AXLN employs a decoupled training-inference scheme in which inter-response correlations are exploited as an auxiliary loss during training. The Hybrid Model learns AXLN-based residuals on top of an MVDA backbone. Four models were compared on two pharmaceutical DoE datasets (Dataset A, n = 32; Dataset B, n = 15). The Hybrid Model achieved the lowest test MAE across all responses (0.551-2.514), significantly outperforming MVDA, AXLN, and ANN (p < 0.05). In the design space comparison, the Hybrid Model correctly excluded an infeasible point that MVDA had erroneously included. The resulting boundary was more reliable from a quality-assurance standpoint. These results show that the proposed Hybrid MVDA-AXLN Model improves both predictive precision and design space reliability within the pharmaceutical QbD framework.
Cefepime is recommended for treating severe pulmonary infections caused by P. aeruginosa using intravenous route that limits sufficient deposition in the bronchi and alveoli of the lungs. To address this, we developed a cefepime dry powder while enabling lower total administered doses using an inhaler. A factorial design (Xn + 1) with two additives, L-leucine and L-tryptophan, was used to prepare inhalable and physicochemical stable powders via spray drying. The preparation parameters retained drug contents above 90 % across all cefepime powder formulations (CPF; n = 10). The particles were spherical with a mean geometric particle diameter 1-2 µm. Leucine significantly improved the aerodynamic properties by increasing the number of dimples on the particles' surfaces. The optimized formulation, CPF-3 (10 % leucine-only), produced the highest fine particle dose (FPD), corresponding to a fine particle fraction (FPF) of approximately 80 %. The minimum inhibitory concentrations (MIC) and 90 % minimum biofilm inhibitory concentrations (MBIC90) of the formulations were comparable to those of the cefepime raw material and safe on the lung epithelial cell line (A549). The powder exhibited moisture-sensitivity and therefore requires storage in a dry environment at room temperature. Overall, this study establishes a robust formulation strategy of inhalable fourth generation β‑lactams.
Chronic neuropathic pain, arising from somatosensory system dysfunction, presents significant therapeutic challenges due to pathogenic complexity, individual variability, and limitations of current pharmacotherapies. Needle-based interventions, characterized by minimal invasiveness, dual analgesic-neurorestorative capabilities, and enhanced patient compliance, offer promising alternatives. However, there is a lack of comprehensive reviews that synthesize both technical advances and clinical applications of needle-based interventions for chronic neuropathic pain. This review synthesizes researches on needle-based chronic neuropathic pain management, categorizing needle-based devices into single needles and needle arrays. We analyze material properties, fabrication methods, therapeutic mechanisms, and clinical efficacy across major chronic neuropathic pain conditions (sciatica, trigeminal neuralgia, postherpetic neuralgia, diabetic neuropathy, spinal cord injury). Moreover, this review evaluates existing challenges and promising prospects that may accelerate clinical translation.
Pharmaceutical administration studies determine whether a medicine can be prepared, held, and delivered without unacceptable loss of quality. Their conclusions are essential to product instructions, yet the underlying evidence is difficult to use beyond the product and conditions originally tested. This letter contrasts that pattern with physiologically based pharmacokinetic modelling, a field in which system, drug, and study information are represented separately and the purpose and limits of model use are made explicit. Administration science does not require a PBPK model. It does, however, need records that preserve the relationships among product and formulation attributes, administration-system components, preparation and use conditions, measured outcomes, and the boundaries of the experiment. Standardized, structured records would not remove the need for product-specific studies. They would make previous work easier to interpret, reveal when further testing is necessary, and allow administration evidence to contribute to cumulative pharmaceutical knowledge.
Co-amorphous systems (CASs) effectively enhance the solubility of poorly soluble drugs, but their development is often limited by the scarcity and inadequate co-amorphization capability of existing co-formers, resulting in poor physical stability and limited clinical applicability. Therefore, identifying new co-formers is essential. In this study, 18β-glycyrrhetinic acid (GA), a natural licorice-derived sweetener, was evaluated as a co-former for CAS development using basic albendazole, neutral carbamazepine, and acidic indomethacin as model drugs. GA exhibited excellent amorphization capability, achieving complete amorphization within 30 min of cryogrinding. In the presence of GA, all three drugs were fully amorphized within a short cryogrinding time, whereas amorphization remained incomplete after 5 h without GA. Modulated differential scanning calorimetry confirmed homogeneous CAS formation, with elevated glass transition temperatures (93.46-114.43 °C), while Fourier transform infrared spectroscopy revealed strong intermolecular interactions. All CASs exhibited higher maximum drug concentrations than their crystalline counterparts in supersaturated dissolution tests. In addition, no recrystallization was observed after storage under 75 % relative humidity for 6 months or dry conditions for 12 months at 25 °C, demonstrating excellent physical stability. These findings identify GA as a promising co-former for expanding the design space of CASs.
Chronic hepatitis B virus (HBV) infection is characterized by an inadequate immune response to clearing the virus. Immunomodulatory agents present a promising therapeutic strategy for the treatment of HBV. Glucan particles (GPs), when used as a vaccine adjuvant, have shown the ability to enhance both innate and adaptive immune responses. Thus, we assessed the effectiveness of the GPs-based hepatitis B vaccine in boosting the immune response against HBV. Through biodistribution studies in mice, we confirmed GPs liver accumulation after oral administration. The investigation of GP-hepatocyte interactions revealed that GPs induce elevated production of reactive oxygen species (ROS) in HepG2 cells without affecting cellular glutathione levels. Additionally, GPs promoted the maturation of human dendritic cells (CD80, CD86, MHC-I and MHC-II). Vaccination studies in mice with GPs encapsulating HBsAg, HBcAg, and CL097, a TLR7 agonist, elicited a robust cellular immune response, characterized by enhanced cytotoxic T cell functions, cytokine production, and spleen cell proliferation. Moreover, high levels of serum anti-HBsAg IgG and antigen-specific fecal sIgA were also induced. Notably, the GP-based vaccine also enhanced IFN-γ in the liver. These findings highlight the potential of oral administered GPs for liver targeting and their immunostimulatory properties in the context of hepatitis B vaccination.
Precise targeting of antigen-presenting cells (APCs) via mannose is a classic vaccine strategy, but traditional random conjugation often fails to mimic natural pathogen glycan arrays, limiting receptor cross-linking and risking immune tolerance. This review systematically decodes the critical geometric thresholds of nanointerfaces regulating mannose targeting. We elucidate how optimal ligand spacing and density synergistically trigger the 'glycosyl cluster effect' for efficient receptor aggregation, contrasting the biophysical recognition between rigid polymers and fluid lipid nanoparticles (LNPs). To transform targeted uptake into potent adaptive immunity, we further analyze post-endocytosis intracellular transport. We emphasize coupling interface recognition with pH-responsive endosomal escape mechanisms to rescue antigens from lysosomal degradation, redirecting them toward MHC-I cross-presentation for robust CD8 + T cell responses. Finally, addressing field fragmentation, we innovatively propose the 'Minimum Information About a Carbohydrate Agent (MIACA)' framework for glycan-based nano-carriers, to mandate the evaluation of effective ligand density, binding kinetics, and spatial bioavailability, providing an engineering blueprint for clinically translatable precision vaccines.
The oral delivery of biologics such as oligonucleotides may be enabled by the use of a chemical permeation enhancer such as sodium caprate (C10). However, C10 has low solubility under acidic gastric conditions which, often combined with poor drug stability, limits absorption when formulated in oral formulations. In this work, we investigated the use of alkalizers in immediate release C10-based formulations to enable the delivery of an oligonucleotide drug, RO7062931. Across the physiological pH range, C10 demonstrated low solubility and RO7062931 demonstrated poor chemical stability at pH values lower than 5, which together present significant obstacles to the oral delivery of the oligonucleotide. Screening alkalizers revealed that these pH-modifying components rapidly raised both the simulated gastric fluid bulk pH and microenvironmental pH when combined with C10, increasing the amount of C10 dissolving. Incorporating the most promising alkalizer, sodium carbonate, into C10-containing tablets of RO7062931 led to improved oligonucleotide chemical stability and C10 solubility under gastric conditions during in vitro testing, with the most effective formulation exhibiting complete RO7062931 stability and full C10 release. Alkalizer addition further contributed to oligonucleotide/C10 co-release, which is important for maximizing permeation enhancement activity at epithelial barriers. These findings offer insight into alkalizer incorporation as a promising strategy to improve the gastric delivery of oral formulations of oligonucleotide drugs and permeation enhancers.
Skin aging is a complex biological process driven by collagen degradation and oxidative damage, often exacerbated by UV exposure. While resveratrol is a premier antioxidant for anti-aging, its topical efficacy is historically limited by poor water solubility and weak skin penetration. This study addresses these barriers by developing optimized polyethylene glycol-based liposomes (PEG-liposomes) as novel deformable liposome derivative. The uncoated vesicles demonstrated high stability and uniformity, with a size of 320.17 ± 0.25 nm and a zeta potential of -35.2 ± 0.4 mV. By coating these vesicles with 0.1% hyaluronic acid (HA), successful surface functionalization and improved size stability was achieved. Ex vivo and confocal studies confirmed that these PEG-liposomes successfully reached the deep dermis; notably, a sequential application strategy of the coated and uncoated PEG-liposomes showed that uncoated vesicles could transiently disrupt lipid structures to facilitate deeper penetration for the HA-coated versions. When integrated with micro-needling via a derma roller, the system triggered significant collagen remodeling and a surge in antioxidant enzymes like SOD, while simultaneously reducing oxidative stress markers such as MDA, MMP-1, and the aging indicator beta-galactosidase. The cooperative effect of this dual-vesicle system not only restored the structural integrity of the dermal-epidermal junction but also significantly outperformed existing commercial resveratrol products. Our findings demonstrate that this novel PEG-liposomal system provides a transformative approach to deep-tissue rejuvenation. Through its unique synergistic mechanism with micro-needling, it achieves biological remodeling and dermal restoration that surpasses the efficacy of current market-leading standards.
Cancer is still a leading cause of mortality worldwide and thus more effective and less toxic therapeutic regimens are constantly required. Modern chemotherapeutic agents commonly suffer from a narrow therapeutic window, the potential for severe adverse drug reactions, and lack of metabolic stability which can lead to either decreased exposure at the target site or selective exposure to toxic metabolites. Deuteration is the process of replacing hydrogen (1H or H) with deuterium (D), a heavy and stable isotope of hydrogen, which creates a unique chemical avenue to enhance the bioavailability, pharmacokinetics (PK) and safety profile of both new and existing anticancer agents. This improvement is mainly due to the kinetic isotope effect (KIE), which significantly augments C - D bonds in comparison with C - H bonds, and hence retards enzymatically catalyzed oxidative metabolism by CYP enzymes. This review summarizes the key principles of deuteration, discusses its use in increasing the potency and decreasing toxicity of drugs (including a flavonoid apigenin, as well as several clinical candidates), and describes recently emerging applications of Deuterium Magnetic Resonance Imaging (dMRI) for non-invasive assessment of early tumor response to therapy. We finish with the advanced state of some deuterated anticancer drugs in preclinical and clinical development supporting that pharmacologic deuteration is a potent approach to exploiting certain drug metabolism pathways for improving oncology therapy.
