Frontiers in Pharmacology最新文献

Membrane proteins govern essential cellular processes, including ion transport, signal transduction, and molecular recognition, and collectively represent more than half of all current therapeutic targets. Yet their structural characterization remains challenging due to intrinsic instability, amphipathic surfaces, and conformational heterogeneity. Over the past decade, antibody-based approaches, spanning full-length immunoglobulins, antigen-binding fragments (Fabs), nanobodies, and engineered scaffolds such as designed ankyrin repeat proteins (DARPins), have transformed structural biology by stabilizing dynamic states, augmenting molecular weight for cryo-electron microscopy (cryo-EM), and enabling visualization of previously inaccessible complexes. In parallel, advances in artificial intelligence and machine learning have begun to enhance predictive modeling, accelerate structure determination, and guide rational design of protein-ligand and antibody-antigen interactions. This review examines how antibody engineering and AI-driven computation together are reshaping the landscape of structural biology and therapeutic discovery.

Introduction: Partial bladder outlet obstruction (pBOO) is the most common cause of lower urinary tract symptoms (LUTS). Prolonged BOO induces bladder remodeling, which can lead to severe bladder dysfunction and refractory LUTS in some patients, even after obstruction resolution. This condition significantly impairs patients' quality of life, and no effective treatment is currently available. This study investigated a pBOO rat model using IR-780, a novel near-infrared lipophilic dye with potential targeted antioxidant effects.

Methods: A partial ligation of the rat bladder neck was performed to establish a pBOO model. After confirming successful modeling, the rats were randomly divided into sham, sham + IR-780, pBOO, and pBOO + IR-780 groups (eight rats per group). One week post-surgery, rats received intraperitoneal injections of IR-780 (0.667 mg/kg) or an equivalent volume of phosphate buffered saline solution twice weekly for 3 weeks. Before evaluating efficacy using the bladder filling manometry method, we examined the distribution of IR-780 in tissues and subcellular compartments via confocal fluorescence imaging.

Results: IR-780 accumulated at high levels in the bladders of rats with pBOO, where it was primarily taken up by bladder smooth muscle cells (BSMCs) and localized within the mitochondria. Bladder pressure measurements revealed that IR-780 significantly improved bladder function in rats with pBOO. IR-780 effectively mitigated pathological changes in bladder smooth muscle tissue and concurrently alleviated pBOO-induced reflux nephropathy. In vitro and in vivo experiments confirmed that IR-780 significantly reduced apoptosis in BSMCs. Moreover, cryosection staining and transmission electron microscopy results demonstrated that IR-780 markedly decreased reactive oxygen species levels in BSMCs from rats with pBOO, prevented mitochondrial mass and morphological damage, and significantly reduced the levels of mitochondrial apoptosis pathway-related proteins (Bcl-2, Bcl-2-associated X, cytochrome C, and Caspase-9). We found that IR-780 upregulated the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and its associated antioxidant proteins in the bladder tissue of rats with pBOO.

Conclusion: IR-780 improved urinary function and complications in rats with pBOO by protecting BSMC mitochondria from oxidative stress, which was potentially mediated through the activation of the Nrf2 pathway.

Background: Accurate evaluation of regional drug deposition within the respiratory tract is essential for optimizing inhalation therapy efficacy and minimizing adverse effects. However, non-invasive, real-time quantitative methods for site-specific drug delivery assessment remain limited.

Objective: To develop mathematical models to predict site-specific drug deposition from a dry powder inhaler (DPI) using a non-invasive, real-time photo reflection method (PRM).

Methods: Using Symbicort^® Turbuhaler^® as a model DPI, four inhalation patterns varying in peak flow rate (PFR: 30-60 L/min) and flow increase rate (FIR: 3.2-9.6 L/s²) were simulated using a human inhalation flow simulator. Aerodynamic particle deposition of budesonide was quantified as the fine particle fraction for the whole lung (FPF_WL), peripheral airways (FPF_PA), and oropharyngeal region using an Andersen Cascade Impactor. Particle emission signals were monitored via PRM. The relationship between particle emission signals and deposition performance was analyzed using four univariate models: linear, logarithmic, Hill, and Emax.

Results: Increased PFR and FIR enhanced drug deposition in both the lungs and oropharyngeal region. FPF_WL and FPF_PA were strongly correlated with total particle emission intensity over time with the Hill model (R ² = 0.86 and 0.74 for FPF_WL and FPF_PA, respectively), reflecting nonlinear deagglomeration. Oropharyngeal deposition correlated with flow rate at particle emission peak, fitting a linear model (R ² = 0.82), consistent with inertial impaction mechanisms.

Conclusion: Using an in-vitro model, particle emission signals enable the prediction of site-specific drug deposition from DPI, providing non-invasive, real-time indices and offering personalized inhalation performance assessment beyond conventional flow rate metrics.

Quercetin has attracted increasing attention in the research of treatments for rheumatoid arthritis and osteoarthritis due to its excellent anti-inflammatory, antioxidant, and joint-protective effects. However, the bioavailability of quercetin is relatively low, primarily due to its poor solubility, rapid metabolism, and high clearance rate. Nanotechnology offers new opportunities to enhance the bioavailability of quercetin. Nanoformulations possess many advantages, such as a larger surface area, which can effectively improve the solubility of quercetin. Encapsulating quercetin in nanocarriers can prolong its residence time in the body, thereby improving its bioavailability and therapeutic efficacy. Targeted delivery can be achieved by modifying the surface of nanocarriers with ligands, allowing for direct transport of the drug to the site of injury and increasing the concentration at the lesion site. Nanocarriers can improve drug targeting and release behavior by adjusting their surface properties, thereby enhancing therapeutic effects. This article first summarizes the general preparation methods, drug-loading approaches, and common nanoformulations for quercetin. It then lists and summarizes the applications of quercetin nanoparticle formulations in rheumatoid arthritis and osteoarthritis. Finally, it concludes with a summary and outlook on the clinical applications and challenges associated with quercetin nanoformulations.

Background: Sepsis-induced cardiomyopathy (SIC) is a severe complication of sepsis that markedly increases mortality. Owing to its complex pathogenesis, no targeted drugs for SIC is currently available, highlighting the need for preventive interventions. This study aimed to evaluate whether early administration of Shenfu Injection (SFI) could prevent SIC.

Methods: Patients diagnosed with sepsis 3.0 upon intensive care unit (ICU) admission but without SIC were randomly assigned to either the SFI group or control group via envelope randomization. The SFI group received intravenous SFI (200 mL/day) in addition to standard sepsis or septic shock management for 7 days (minimum 72 h if discontinued due to ICU transfer or death). The control group received an equivalent volume of saline alongside standard care. The primary outcome was the incidence of SIC within 7 days.

Results: A total of 152 patients (76 per group) were analyzed. The incidence of SIC within 7 days was 9.2% in both groups. In the generalized linear mixed model (GLMM) adjusted for gender, age, septic shock and the baseline value of N-terminal pro-B-type natriuretic peptide (NT-proBNP), the interaction between time and group had a significant effect on NT-proBNP levels (P = 0.004). No significant differences were observed between groups in hemodynamic parameters, immune inflammatory indicators, organ function, vasoactive drug use, 7-day fluid balance, 28-day mortality, duration of mechanical ventilation, continuous renal replacement therapy duration, ICU stay, or total hospital stay.

Conclusion: Early application of SFI did not significantly reduce the incidence of SIC in ICU patients with sepsis.

Clinical trial registration: [https://www.chictr.org.cn/], identifier [ChiCTR2400088766].

Background: Radiation-induced lung injury (RILI) is a major dose-limiting toxicity in thoracic radiotherapy, and accumulating evidence implicates radiation-induced cellular senescence in its pathogenesis. This study aimed to investigate whether combination therapy with dasatinib and quercetin (DQ) could mitigate RILI by reducing senescent cell burden.

Methods: A rat model of RILI was established using a single 30 Gy irradiation to the right lung. Pulmonary pathological changes, fibrosis, DNA damage, and cellular senescence were assessed by histology, immunofluorescence, senescence-associated β-galactosidase staining, Western blotting and immunohistochemistry. Transcriptomic profiling was performed to explore the underlying molecular mechanisms.

Results: Compared with irradiation alone, DQ treatment significantly alleviated radiation-induced inflammatory cell infiltration and collagen deposition, reduced γH2AX levels, decreased senescence-associated markers p53, p21, and p16, and suppressed multiple senescence-associated secretory phenotype (SASP) factors. Transcriptomic analysis indicated that DQ-mediated effects were closely associated with activation of apoptotic pathways and modulation of p53, MAPK, PI3K-Akt and mitophagy signaling cascades.

Conclusion: DQ attenuated RILI in rats, with effects consistent with the reduced radiation-induced senescent cells and suppression of senescence-associated inflammatory responses.

Introduction: Vulvovaginal candidiasis (VVC) is one of the most common fungal infections requiring more effective and patient-friendly therapies. This study introduces repurposed Ketoprofen (KPN) Quatsomes (QS) as a novel nano-platform for localized antifungal treatment.

Methods: KPN-QS were prepared using quaternary ammonium surfactants and cholesterol via probe sonication and optimized through a 3¹ × 2² mixed factorial design using Design-Expert^® software. The effects of quaternary ammonium surfactant type (factor A), amount of vesicle-forming materials (factor B), and cholesterol-to-surfactant ratio (factor C) were evaluated to maximize entrapment efficiency and zeta potential, while minimizing particle size.

Results: The optimized QS exhibited spherical nano-sized vesicles (113.7 nm) with high entrapment efficiency (96.8%) and strong positive zeta potential (72.5 mV), ensuring stability and enhanced mucosal adhesion. TEM confirmed the spherical morphology, and in vitro release showed biphasic behaviour with 86.5% release after 8 h, alongside excellent storage stability. Repurposing KPN as an antifungal agent significantly enhanced both in vitro and in vivo microbiological efficacy. The formulation displayed promising MIC values against Candida albicans and markedly improved antifungal performance in vivo VVC model. The KPN-QS group exhibited 4.807 and 2.941 log reductions in fungal count compared to the negative control and KPN suspension, respectively, with complete eradication in three rats after 72 h. Histopathological analysis confirmed the safety of QS on vaginal mucosa.

Conclusion: Collectively, repurposed KPN-QS constitute a stable, biocompatible nanocarrier for targeted vaginal delivery, demonstrating superior antifungal activity and therapeutic potential in VVC.

Epigenetic modifications on nuclear and mitochondrial DNA constitute key regulatory layers influencing the transcriptional, metabolic, and phenotypic adaptability of cancer cells. The canonical principles of epigenetic control encompass DNA methylation, histone modification, and non-coding RNA-mediated regulation, which collectively contribute to the silencing of tumor suppressor genes, the activation of oncogenes, and chromatin remodeling. Therefore, epigenetic drugs (epi-drugs) are of great interest in the development of new-generation therapeutics and holistic treatment approaches. Accordingly, this work presents a narrative review that integrates current evidence on the molecular mechanisms, therapeutic developments, and translational relevance of epigenetic and mitoepigenetic regulation in cancer. RNA-mediated regulation collectively contributes to the silencing of tumor suppressor genes and to the activation of oncogenes. The field of mitoepigenetics encompasses mitochondrial DNA (mtDNA) methylation, RNA modifications, and post-translational regulation of mitochondrial proteins such as TFAM, DNMT1, and sirtuins, which influence oxidative phosphorylation, redox balance, and apoptotic pathways, thereby affecting tumor initiation, progression, and treatment response. Recent advances in epigenetic drug development include FDA-approved DNMT and HDAC inhibitors and newer agents targeting EZH2, IDH1/2, and DOT1L, which broaden the scope of precision oncology. In addition, modulation of mitochondrial epigenetic mechanisms has been identified as a potential approach for addressing metabolic reprogramming and therapeutic resistance in cancer. The convergence of nuclear and mitochondrial regulatory frameworks reveals the critical need for biomarker-informed, combinatory, and organelle-targeted therapeutic approaches to sustain treatment efficacy. Comprehensive characterization and pharmacological targeting of epigenetic and mitoepigenetic networks provide a structured basis for developing personalized and metabolism-informed interventions in cancer therapy.