Probiotic therapy is an emerging non-antibiotic strategy for bacterial vaginosis (BV). However, the susceptibility of probiotics to environmental factors leads to poor storage stability, and the complex, multi-step processes involved in current protection strategies increase production costs, thereby limiting their further clinical application. Here, we report a single-pot co-lyophilization formulation (termed prebiotic-matrix protective stabilization, PMPS), in which probiotics are uniformly dispersed within a protective prebiotic matrix. The formulation was characterized for microstructure, storage stability, and in vitro antibacterial activity. Furthermore, it was incorporated into a thermosensitive gel for in vivo efficacy evaluation. The probiotic formulation via PMPS method exhibited uniform dispersion by scanning electron microscopy and confocal laser scanning microscopy. It retained >90% viability after 30 weeks at 4 °C under air (vs. ∼20% within 3 weeks for bare probiotic powders). The formulation achieved >99% inhibition of Gardnerella vaginalis with post-co-culture viable counts exceeding 10[9] CFU/mL. Furthermore, the thermosensitive gel system showed superior in vivo efficacy to commercial products. The PMPS method integrates physical protection with nutritional support, enabling long-term storage stability through a simple and cost-effective preparation process, and offers a promising platform for vaginal microecological intervention.

Drug development has long been constrained by high costs, lengthy timelines, and low success rates, primarily due to the insufficient human physiological fidelity of traditional preclinical models. Artificial intelligence (AI) has accelerated computational drug discovery, but its models inherit training-data bias and are rarely validated against human-relevant experiments. Organ-on-a-chip (OoC) systems produce human-relevant, multimodal physiological data, yet the sheer volume of data they generate overwhelms conventional processing pipelines. With regulators now encouraging non-animal testing, integrating AI with OoC has become a practical route forward. This review examines how the two technologies reinforce each other across the OoC lifecycle, surveys progress toward clinical and industrial translation, and considers longer-term paradigms such as digital twins and programmable virtual humans. Major hurdles remain in hardware performance, biological fidelity, data standardization, and model interpretability. Closer collaboration across sectors would help this combined dry-wet approach reduce animal testing and make drug research more predictive and more human-relevant.

Several studies have measured and rationalised how dissolution medium pH alters the drug crystal surface pH and dissolution rates for ionisable compounds, with predictive models proposed. However, mechanistic understanding of buffer species effects remains limited and existing models are often complex. Other extrinsic parameters including temperature, agitation, and micelles, have received comparatively little attention. This study investigated the impact of parameters on the intrinsic dissolution rate (IDR) of acalabrutinib drug substance, using the rotating disk apparatus. Experiments spanned agitation rates (25, 50, and 100 RPM); temperatures (25 and 37 °C), buffers (sodium acetate, sodium citrate, potassium hydrogen phthalate, sodium phosphate); and micelle-containing media, including FaSSIF-V2, FeSSIF, and sodium dodecyl sulfate (SDS) in pH 6.8 phosphate buffer at varying concentrations. A simple mathematical model, that combines the estimation of surface solubilities (through the surface pH), and a direct calculation of the IDR has been employed to rationalise the experimental findings and to achieve a better mechanistic understanding of the impact of the buffer media and surfactant effects. The model confirmed the importance of explicitly including surface pH in the estimation of surface solubilities, however, lacked predictive capability and generalisability for a quantitative prediction of the IDR in buffered media. The requirement for the fitting of system-dependent diffusivities suggests that a detailed description of transport phenomena for charged species is needed to capture the impact on the IDR per buffer. The transfer of the mechanistic understanding to inform a physiologically based biopharmaceutics model (PBBM) is also discussed.

The therapeutic efficacy of imatinib (IM) in gastrointestinal stromal tumors (GISTs) is often limited by inadequate intratumoral drug accumulation and an abnormal tumor microenvironment. In this study, an ultrasound-responsive poly(lactic-co-glycolic acid) (PLGA) nanoparticle system, designated CXB/CAT@PFP-PLGA NPs, was developed for the co-delivery of celecoxib (CXB) and catalase (CAT) to enhance the antitumor efficacy of IM. The nanoparticles were characterized in terms of their physicochemical properties, ultrasound responsiveness, and oxygen-generating capability. Their ability to potentiate IM therapy was evaluated both in vitro and in vivo, while biodistribution and biosafety were assessed in vivo. The results showed that CXB/CAT@PFP-PLGA NPs exhibited a relatively uniform size distribution and successfully co-encapsulated CXB and CAT. Upon ultrasound irradiation, the nanoparticles underwent phase transition, thereby triggering drug release. In vitro, CXB/CAT@PFP-PLGA NPs enhanced the inhibitory effect of IM by reducing cell viability, promoting apoptosis, and inhibiting cell migration. In vivo, the nanosystem improved the antitumor efficacy of IM, accumulated at the tumor site, suppressed tumor growth, and showed no evident systemic toxicity. Collectively, these findings suggest that CXB/CAT@PFP-PLGA NPs may represent a promising strategy for enhancing IM therapy in GIST.

The aqueous solubility and membrane permeability of Biopharmaceutics Classification System (BCS) class II and IV drugs, such as furosemide (FUR), sulfadiazine (SDZ), verapamil (VEP), mefloquine (MEF), and the bioactive compound quercetin (QUE), are low. Thus, affecting the oral bioavailability of these drugs. In our study, we aimed to design drug-modified low-generation poly(amidoamine) (PAMAM) dendrimer complexes to improve solubility and permeability. The PAMAM dendrimers (G0 and G1) were synthesized via a divergent approach that combined Michael addition and amidation reactions, using 1,4-diaminobutane as the core. Drug-dendrimer complexes were prepared at molar ratios of 1:1, 3:1, and 5:1 by co-dissolution in methanol, followed by solvent removal and aqueous extraction. The reduction in crystallinity was classified using a polarized light microscopy-based scoring system ranging from CCCC (fully crystalline) to AAAA (fully amorphous), enabling qualitative assessment of amorphization following dendrimer incorporation. Polarized light microscopy revealed complete amorphization (AAAA) in specific systems at a 1:1 molar ratio. Solubility studies at pH 1.2, 6.8, and 7.4 showed significant increases, with quercetin reaching nearly 5-fold higher concentrations for the PAMAM-G1 1:1. Parallel Artificial Membrane Permeability Assay (PAMPA) experiments showed variable permeability responses across drug: dendrimer ratios and pH conditions. PAMAM dendrimer complexes were able to considerably enhance the solubility of BCS class II and IV drugs through amorphization and non-covalent interactions. We expect that these systems will further improve key biopharmaceutical properties directly associated with the bioavailability of other poorly water-soluble drug candidates.

Wet granulation is a critical unit operation in the manufacturing of solid dosage forms. However, conventional wet granulation techniques are limited by significant batch-to-batch variability and scale-up challenges, making them difficult to meet the demands of continuous pharmaceutical manufacturing and robust quality control. As an emerging continuous granulation technology, Twin-Screw Wet Granulation (TSWG) has attracted extensive attention in pharmaceutical research and industrial applications due to its advantages of continuous processing, modular design, controllable mixing and shear, and reduced scale-up effects. This review systematically summarizes and critically evaluates the core technologies and underlying mechanisms of TSWG, including equipment configuration, screw design, and granulation mechanisms. The relationships between formulation variables, equipment parameters, and process conditions with critical quality attributes (CQAs) of granules, such as particle size distribution, porosity, and mechanical strength, are comprehensively analyzed. Furthermore, the application of Process Analytical Technology (PAT) for real-time monitoring is discussed, together with recent advances in process modeling and optimization using Discrete Element Method-Computational Fluid Dynamics (DEM-CFD) coupled models, Population Balance Models (PBMs), Machine Learning (ML), and Artificial Intelligence (AI). The potential of integrating AI with PAT, machine vision, and data-driven models for granule quality prediction, online monitoring, and intelligent process control is also highlighted. Finally, the current challenges of TSWG are addressed, including process scale-up consistency, adaptation to raw material batch variability, and control of active pharmaceutical ingredient (API) polymorphic stability. Future development is expected to rely on interdisciplinary integration to advance technological innovation and support Quality by Design (QbD) paradigm in pharmaceutical manufacturing.

Pancreatic ductal adenocarcinoma (PDAC) is driven by KRAS mutations in over 90% of cases yet remains refractory to most therapies due to poor antigen delivery and a suppressive tumor microenvironment. To address these challenges, we developed an exosome‑augmented, epitope‑focused mRNA nanovaccine. First, we engineered a series-connected (SC) mRNA that condenses five immunodominant KRAS‑mutant epitopes (G12D, G12V, G12R and two other sequences) into a single open reading frame. This design minimizes non‑productive sequence and enhances antigen presentation compared to a conventional parallel-connected (PC) mixture of full‑length transcripts. Second, we cloaked β‑sitosterol LNPs with mature dendritic‑cell-derived exosomes (LNP@exo), endowing the particles with lymph‑node tropism and intrinsic adjuvanticity. The resulting mKRAS SC-LNP@exo triggered potent dendritic cell activation and Th1 cytokine release in vitro, rapidly accumulated in lymph nodes, and drove superior CD8[+] T cell infiltration in a Pan02 tumor model. Remarkably, one‑third of mKRAS SC-LNP@exo treated mice achieved complete tumor regression without off‑target toxicity. These findings demonstrate that combining epitope‑condensed mRNA with exosome‑cloaked LNP delivery can convert "cold" KRAS‑mutant PDAC into an immunologically responsive tumor and provide a broadly applicable strategy for next‑generation mRNA cancer vaccines.

Mupirocin is a commonly used broad-spectrum topical antibiotic. However, its therapeutic applications are limited by its short half-life, necessitating frequent administration. A biopolymer-coated, layer-by-layer (LbL)-assembled mupirocin microsphere (LbL-MUP)-based wound dressing was developed to provide sustained drug release and a moist wound environment. Core alginate-based mupirocin microspheres were prepared by ionotropic gelation and sequentially LbL-coated with oppositely charged biopolymers, chitosan/gellan gum. Based on physicochemical and functional properties, F4 with chitosan/gellan gum (4:1) was considered the optimized batch. The optimized LbL-MUP formed an interconnected polymeric network, exhibiting cumulative sustained release of 91.03 ± 1.80 over 24 h. LbL-MUP exhibited a fluid uptake capacity of 477.66 ± 3.82%. These in-situ microspheres exhibited a thermosensitive transition to a high-viscosity gel at physiological temperatures, thereby effectively absorbing wound fluid. SEM micrographs of LbL-MUP showed a rougher surface than uncoated microspheres. FTIR study indicated polymer deposition; XRD and DSC revealed the amorphous nature of the formulation. LbL-MUP showed superior antimicrobial efficacy against S. aureus and Escherichia coli than mupirocin. LbL-MUP accelerated wound healing than marketed formulation, achieving 96.79 ± 2.09% wound contraction by day-21 in rats. Histopathology confirmed complete re-epithelialization and normal collagen deposition, while biomarker estimation showed TGF-β1 (926.69 ± 10.77 pg/mL) on Day-5, important for initiating wound closure. LbL-MUPs can be sprinkled onto an infected wound with minimal contact, transforming into a gel-based wound dressing that provides sustained release and maintains a moist wound environment. Thus, LbL-MUP highlights its potential as a scalable biopolymer-based formulation for the topical management of infected wounds, which may help address challenges in wound management and antimicrobial resistance.

Targeted protein degradation (TPD) has transitioned from a paradigm-shifting concept to a clinically validated strategy, with multiple degraders achieving proof-of-concept in oncology and a rapidly expanding toolbox. By co-opting the ubiquitin-proteasome system to eliminate pathogenic proteins in an event-driven manner, TPD addresses targets long deemed undruggable. To capture this momentum, we present a comprehensive overview of the field. We dissect the mechanisms of heterobifunctional PROTACs and their clinical translation, covering late-stage programs targeting canonical oncogenic drivers (AR, ER, BTK) and emerging first-in-human studies against historically challenging targets such as STAT3 to illustrate both clinical validation and mechanistic expansion. Beyond classical PROTACs, we systematically summarize the expanding TPD toolbox: lysosome-targeting chimeras for degrading extracellular and membrane proteins, autophagy-based degraders for clearing aggregates and damaged organelles, antibody- and nucleic acid-derived PROTAC formats for tissue-specific delivery and transcription factor targeting, deubiquitinase-targeting chimeras for protein stabilization, and proximity-based post-translational modification editing, collectively demonstrating the broad reach of proximity-inducing pharmacology. By integrating molecular mechanisms with therapeutic applications, this review illustrates how TPD is reshaping the druggable proteome. We outline key challenges including novel E3 ligase ligand discovery, tissue selectivity, and acquired resistance, and discuss how covalent fragment screening, artificial intelligence, and expanded E3 ligase repertoires will advance next-generation degraders to fulfill the promise of event-driven pharmacology. This review provides a roadmap for translating TPD into transformative therapies.

Stratum corneum (SC) barrier damage through dermally applied substances with irritation potential is well documented. However, little information about the impact of combined chemical and physical aggressors on human skin physiology in vivo exists. Thus, the aim of this study was to evaluate SC alterations caused by treatment with o/w emulsions stabilized by diacyl and monoacyl phospholipids and control surfactants with and without UVA irradiation. Biophysical skin parameters of eight healthy volunteers were examined on the volar forearm 1 h and 24 h after occlusive treatment with Finn chambers using skin probes to analyze transepidermal water loss (TEWL), pH, hydration, erythema and melanin index. Quantitative confocal Raman spectroscopy (CRS) was employed to non-invasively collect depth-resolved skin spectra. After recovery, the treatment was repeated in combination with 20 J/cm[2] UVA exposure. In addition, ex vivo studies with similar protocol (semi-occlusive treatment) were conducted to complement in vivo data. Results showed increased TEWL and skin surface pH 1 h after sodium lauryl ether sulfate (SLES) emulsion treatment (TEWL: +7.7% to + 10.2%; pH: +16.6% to + 17.5%), accompanied by reduced SC hydration and natural moisturizing factor (NMF) content (hydration: -32.9% to -36.0%; NMF: -27.1% to -27.6%) compared to a diacyl phospholipid emulsion and the controls (water, polysorbate 80 emulsion), irrespective of UVA irradiation. Surprisingly, mono acyl phospholipids had lower impact than anticipated regarding their high emulsifying power, suggesting skin-friendliness when used in emulsion products. Comparative studies ex vivo showed limited potential to detect biophysical differences in human skin observed in vivo.

Accurate measurement of aerosol velocity is important for evaluating the performance of orally inhaled and nasal drug products (OINDPs). This study aims to demonstrate and evaluate the accuracy and reliability of a novel instrument, the "OptoSleeve," which integrates a compact optical diagnostic system designed to measure aerosol velocity and plume duration across various aerosol drug emitting devices. The OptoSleeve employs the laser extinction concept to capture aerosol flow behaviour. Experiments in this study were performed using nasal sprays, pressurised metered dose inhalers (pMDIs), and dry powder inhalers (DPIs) under a range of flow rate conditions, and the results were compared with reference measurements obtained from Particle Imaging Velocimetry (PIV) and Particle Tracking Velocimetry (PTV). The OptoSleeve was shown to provide velocity data that closely align with those obtained using PIV and PTV. Further complementary studies, conducted using computational fluid dynamics modelling, revealed that integrating the OptoSleeve with a regulatory-approved throat replica (e.g., the United States Pharmacopeia throat) has minimal impact on aerosol transport and deposition dynamics within the replica.

Reliable biodistribution assessment is essential for the nonclinical evaluation of plasmid DNA (pDNA)-based therapeutics. Here, we developed and performed fit-for-purpose validation of a TaqMan-based quantitative PCR (qPCR) assay for the absolute quantification of pDNA following electrode-needle device-assisted intramuscular electroporation. The assay showed high target specificity, linear quantitative performance over six orders of magnitude, and acceptable accuracy and precision under standardized genomic DNA matrix conditions. Using this assay, we characterized the biodistribution of the SARS-CoV-2 spike-encoding DNA vaccine pSARS2-S in Sprague-Dawley rats following intramuscular administration with or without electroporation. Plasmid DNA was detected predominantly in the injection-site region, particularly in muscle and overlying subcutaneous tissue, whereas substantially lower levels were observed in non-target organs. At the terminal sampling time point, pDNA levels in the evaluated major organs, including the gonads, were below the lower limit of quantification (LLOQ), while low-level residual pDNA remained detectable in subcutaneous tissue. These findings characterize the tissue distribution and persistence of pSARS2-S under the established delivery condition. Overall, this study establishes a reproducible, fit-for-purpose qPCR assay and analytical workflow for quantitative nonclinical biodistribution assessment of pSARS2-S.

Powder bed fusion-laser beam (PBF-LB) was used to investigate the influence of three Candurin® pigments (Gold Sheen, NXT Ruby Red and Orange Amber, 3 % w/w) on the fabrication of thin dosage forms composed of Parteck® MXP 4-88 and zolpidem tartrate (ZT). The effects of photoabsorber type, model thickness and build orientation were evaluated. Horizontally printed dosage forms generally showed greater dimensional accuracy, whereas vertical printing was associated with larger z-axis deviations and one-sided bending deformation, likely due to the greater number of deposited layers. A statistically significant difference was observed between F Gold and F Ruby, while F Amber exhibited intermediate behaviour, indicating that photoabsorber type may contribute to early dissolution under the investigated conditions. MIP and SEM analyses revealed complex porous structures resulting from partial sintering, suggesting that drug release may reflect the combined influence of multiple microstructural characteristics rather than total porosity alone. Drug assay results indicated no substantial loss of recoverable ZT content, and FTIR analysis showed no evidence of drug-excipient interaction. These findings highlight the importance of photoabsorber selection and build orientation in modulating the internal structure and dissolution behaviour of thin dosage forms, supporting the potential of PBF-LB for future personalised dosing strategies.