European Journal of Pharmaceutical Sciences最新文献

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are severe inflammatory lung diseases with high morbidity and mortality, lacking specific treatments. Inhalation offers direct access to the damaged lung epithelium, making tracheal gene delivery a promising approach. However, challenges such as low efficiency, off-target effects, and repeated dosing limit its effectiveness. In this study, we developed an inhalable recombinant adenoviral vector (Ad-IL4/10) carrying dual reporter genes to deliver interleukin-10 (IL10) and interleukin-4 (IL4) to the lungs, enabling efficient and sustained expression of anti-inflammatory cytokines. Using a lipopolysaccharide (LPS)-induced lung injury mouse model, the anti-inflammatory effects of aerosolized Ad-IL4/10 were evaluated. The results showed that aerosolized Ad-IL4/10 significantly reduced weight loss, lung wet-to-dry weight ratio, and total protein levels in bronchoalveolar lavage fluid (BALF). Additionally, it alleviated pulmonary inflammation and alveolar damage while suppressing proinflammatory markers and LPS-induced monocyte and neutrophil infiltration. Ad-IL4/10 also restored monocyte-macrophage homeostasis. These findings indicate that an inhalable adenoviral vector effectively mitigates LPS-induced lung injury through IL4 and IL10 delivery, offering a promising therapeutic strategy for ALI.

Triple-negative breast cancer (TNBC) remains a highly aggressive disease with limited therapeutic options, and metastasis is the leading cause of patient mortality which highlights a major unmet medical need. Nevertheless, the mechanisms regulating metastatic progression remain poorly understood. Extracellular vesicles (EVs) have recently emerged as key mediators of the intercellular communication. Ubiquitously secreted, these nanoparticles provide a convenient and informative glimpse into cellular physiology given that their molecular composition shall reflect the state of their parent cell. Therefore, in this study, we aim to show that lipid profiles of EVs may differentiate malignant from non-malignant cells and to reveal biomarkers associated with TNBC. For this reason, we investigated the lipid composition of EVs released by cancer cells (MDA-MB-231, MDA-MB-453 and MDA-MB-468 representing TNBC) and non-malignant (MCF10A) human breast epithelial cells, with the goal of highlighting specific lipid profiles that might serve as potential biomarkers for TNBC. EV isolation and characterization were performed in accordance with MISEV2023 guidelines, while lipidomic profiling was carried out using an untargeted liquid chromatography and high-resolution mass spectrometry approach. We annotated over 500 complex lipid species. Multivariate analysis was used to explore major trends in the dataset, separating EVs derived from cancerous and non-cancerous cells and capturing differences among TNBC-derived EVs, reflecting their distinct genetic background. Further supervised modelling revealed distinct TNBC lipidomic signatures including phosphatidylcholines with alkyl chains of C18:0_C19:1, C20:1_C22:5, C18:1_C22:6 or C15:0_C22:5, C17:0_C20:3 and phosphatidylserine C32:0 which consistently emerged as shared lipid species across TNBC-derived EVs, highlighting their potential as biomarkers for the detection of TNBC. Furthermore, we highlighted lipid subsets specific to each of the investigated TNBC's source of EVs for in-depth molecular subtyping of TNBC.

Objectives: To explore the impact of chronological ageing on gene expression levels of tight junction-related genes in the human gastrointestinal (GI) epithelium.

Methods: Biopsies were collected from the epithelium of the stomach, the duodenum, the ascending colon, and the descending colon of adults and of older adults. Relative quantification of gene expression levels for claudin-2, occludin, ZO-1, and JAM-A was performed using real-time qPCR only in biopsies from individuals whose dietary habits, medical record, and endoscopic data did not indicate a potential impact on epithelial barrier function. Relative gene expression analysis was performed in gastric biopsies from 3 adults (39-44 years) and from 3 older adults (68-89 years), in duodenal biopsies from 11 adults (30-45 years) and from 12 older adults (66-89 years), and in paired biopsies from the ascending and descending colon of 6 adults (35-45 years) and 9 older adults (66-87 years).

Results: Expression levels of all four investigated tight junction-related genes in the gastric epithelium, in the duodenal epithelium, and in the epithelium of the ascending and descending colon were not affected by chronological ageing. However, in older adults, the gene expression level of occludin was higher in the descending colon compared to the ascending colon.

Conclusions: Chronological healthy ageing does not appear to impact tight junction-related gene expression levels along the GI epithelium. It would be useful to investigate the barrier of descending vs. ascending colon in older adults with colitis, as the transport of 5-aminosalicylic acid, a frequently used anti-inflammatory agent, occurs, at least partly, via the paracellular pathway.

Tapinarof is a novel aryl hydrocarbon receptor (AhR) agonist that has recently been approved for the treatment of psoriasis. Although its clinical efficacy has been proven, in psoriasis, hyperkeratotic plaques and damaged barrier hinder drug penetration from conventional creams and ointments, resulting in poor bioavailability in the deeper layers of the skin. Innovative drug delivery systems can improve targeted action and reduce potential side effects. The primary goal of this study was to evaluate whether tapinarof-loaded nanogels enhance the anti-inflammatory, anti-proliferative, and anti-migratory effects of tapinarof at the cellular level using an in vitro HaCaT-THP-1 co-culture model. Initially, experiments were performed using real-time cell analysis (RTCA), and a concentration of 10 µM was found to be the most effective, showing significant cell proliferation inhibition compared to imiquimod and free tapinarof. To demonstrate cell proliferation and migration during inflammatory conditions, a wound healing assay was performed, which showed inhibited cell migration. Subsequently, the levels of inflammatory cytokines (TNF-α, IFN-γ, IL-17A, IL-23) were analyzed by ELISA, which demonstrated that tapinarof incorporated into nanogels reduced cytokine production more effectively than the drug alone. Quantitative PCR (qPCR) analysis confirmed Imiquimod (IMQ)-induced upregulation of Tnf-α and Ifn-γ, whereas Il-17a and Il-23 did not show measurable transcriptional changes under the experimental conditions. Finally, inhibition of p65 subunit nuclear translocation were observed during NF-κB pathway activation in THP-1 cells by immunofluorescence staining. Overall, our results indicate that nanogel-based delivery systems exhibit enhanced biological effects of tapinarof compared to the free drug in an in vitro psoriasis-like co-culture model, highlighting their potential as therapeutic tools for the treatment of various inflammatory skin diseases.

Delivering the drug to the eye using topical delivery poses significant challenges, owing to precorneal obstacles, rapid clearance mechanisms, and adverse bioavailability. The recent development of smart lipid-based nanocarriers, including solid lipid nanoparticles (SLNs), nanostructured lipid carriers (NLCs), liposomes, nanoemulsions, and micelles, has demonstrated the capability to address these limitations by increasing mucoadhesion, slowing release, and facilitating penetration of drugs through the epithelial lining. The review examines the categorisation, design concepts, and bio-interfacing strategies of the lipid-based delivery system used in the ocular application. This review focuses on the physicochemical diversity and the capability of traversing the ocular barriers. Several formulations exhibit promising translational applicability, including chitosan-coated SLNs of ofloxacin, which increased transcorneal permeation as compared to commercial drops, and NLCs containing diosmin, which exhibited anti-inflammatory activity and cytocompatibility in in vitro tests against retinal cells. In vivo, ripasudil in liposomes protected fibrosis in the subretinal layer, and NLCs of ciprofloxacin and natamycin enhanced antimicrobial activity with a dual drug combination. Stimuli-responsive systems, such as temperature-sensitive and pH-sensitive hydrogels, offer prolonged ocular residence time and site-specific delivery in the eye. Preclinical studies also support the potential application of SLNs of bimatoprost in the long-term reduction of intraocular pressure and PEGylated NLC in protein delivery in posterior segment diseases. The review has also discussed formulation scalability, toxicity, and regulatory barrier problems, and it also explores future integration with personalised medicine and non-invasive delivery platforms.

Introduction: Seizure disorders are frequent and disabling neurological conditions of the brain that are currently managed mainly with medications. However, the clinically available drugs are fraught with side effects and in some cases ineffectiveness. Thus, there is the need to develop newer and safer anti-seizure medications, especially those based on novel pharmacophores. Here we tested the hypothesis that compounds derived from the oxazolidinone pharmacophore have antiseizure activity in vivo by inducing seizures chemically and electrically in male Sprague Dawley rats and testing their ability to prevent/inhibit these seizures METHODS: Rats were pretreated with 100 mg/kg of each compound (PH066, PH139, PH162, PH166 and PH192) and seizures induced at various time points afterwards by electrical pulse (electrically-induced) and by IP administration of pentylenetetrazol (chemically-induced). Seizure scoring was done visually by two experienced researchers RESULTS: We report here that compound PH162 provided the best protection across all seizure models with the longest duration of action. The rank order of efficacy and duration for all tested compounds is as follows: PH162 > PH139 > PH166 ≈ PH066 > PH192 CONCLUSION: We conclude that although all the tested compounds had some antiseizure activity, PH162 provided a broader based protection with likely more favorable pharmacokinetics. Thus, PH162 is recommended as the lead compound among this group of compounds tested that may be further optimized for even better kinetics and/or safety by slight chemical modification of the basic structure.

Transdermal estradiol patches have been widely used for the prevention and treatment of ovarian insufficiency. However, they should not be used alone and must be combined with adequate progestin therapy for a full course to provide sufficient endometrial protection. This study aimed to develop a compound estradiol-progesterone transdermal patch by incorporating progesterone into the estradiol patch to reduce the number of medications administered, and to evaluate its feasibility and efficacy. Key excipients in the patch, including pressure-sensitive adhesive, permeation enhancers, and plasticizers, were screened and optimized. The optimal patch formulation was selected based on evaluation criteria including adhesion, cumulative release rate, cumulative permeation, and appearance. Subsequently, a menopausal rat model was established to conduct single-dose pharmacokinetic studies and multiple-dose pharmacodynamic evaluations, comparing results with a commercial estradiol patch and progesterone injection. Results demonstrated that the prepared patches exhibited good conformability and adhesion. The cumulative release rate of E2 over 72 hours was 94.58%, with a cumulative permeation rate of 154.56 μg·cm^-2 over 24 hours. These results are comparable to those of commercially available estradiol patches, which exhibit a 72-hour cumulative release rate of 90.02% and a 24-hour cumulative permeation rate of 148.69 μg·cm^-2. P4 exhibited a 72-hour cumulative release rate of 92.45% and a 24-hour cumulative permeation rate of 104.92 μg·cm^-2, achieving the clinically effective dose threshold. The time to peak concentration and residence time for a single application were longer than those of ESTRANA® Tapes and progesterone injection. Compared with the model group, the combined administration group showed a significantly increased uterine coefficient without the abnormal endometrial thickening observed after using estradiol patches alone. This patch achieves synchronized delivery of estrogen and progesterone, meeting the clinical demand for long-acting hormone therapy.

Transcorneal permeation is the main entry route into the eye for topically applied drugs, but the impact of active transport on corneal permeability is still poorly understood. We aimed to identify the influence of transporters for rabbit corneal permeability by utilizing the PermeaSys microfluidic system, which requires less than 20 times smaller tissue pieces and 10 times smaller exposure volumes of the study compound, than the commonly used Ussing chamber. The rabbit corneal permeability was first validated with three model compounds, rhodamine 123, benzoic acid, and paracellular marker compound lucifer yellow, before studying the permeability with three clinically used drugs, ciprofloxacin, diclofenac, and methotrexate. Both bidirectional and inhibition studies were performed. Rhodamine 123 did not show directionality nor altered permeability in the presence of an inhibitor, but the permeability of benzoic acid was affected by a monocarboxylate transporter inhibitor, valproic acid. Clinically used compounds ciprofloxacin, diclofenac, and methotrexate did not show directionality in the corneal permeability studies. However, methotrexate permeability was altered when administrated together with sulfasalazine and MK-571, which can inhibit several known drug transporters. The data generated here with the PermeaSys instrument is comparable to studies conducted with the Ussing chamber and can be reliably used for the study of permeability and active transport in the cornea. However, active transport does not appear to have a great impact on the corneal permeability of the studied drug compounds.

Due to the lack of effective drug therapies, aortic dissection is associated with extremely high mortality rates. Previous studies have demonstrated that FOS-like antigen 1 (FOSL1) plays a role in atherosclerosis and tumor progression. However, the mechanism of FOSL1 in AD remains largely unknown. Therefore, in this study, we aimed to clarify the potential mechanism of FOSL1 in AD and provide a theoretical basis for clinical applications. AD is a life-threatening cardiovascular emergency characterized by a tear in the inner layer of the aorta, resulting in separation of the wall layers and associated with high morbidity and mortality. Understanding the molecular mechanisms driving AD is essential for developing targeted therapies. This study explores the role of the transcription factor FOSL1, known for its involvement in stress response, fibrosis, and cellular differentiation, in AD pathogenesis, with specific focus on collagen deposition and fibroblast-to-myofibroblast transition. Understanding the molecular mechanisms underlying AD is critical for developing novel therapeutic strategies. This study aimed to investigate the role of FOSL1, a transcription factor implicated in cellular differentiation, stress response, and fibrosis, in the pathogenesis of AD and its relationship with collagen deposition and fibroblast phenotype transformation. The role of FOSL1 was investigated by analyzing the Gene Expression Omnibus dataset and evaluating in vitro and in vivo models. Immunohistochemistry and Immunofluorescence assays were used to determine the functional localization of FOSL1 in cells. The effect of FOSL1 expression levels on the efficacy of Naringenin in treating AD was analyzed through combined in vivo and in vitro Naringenin experiments. FOSL1 expression was upregulated in AD, and FOSL1 promoted the proliferation of aortic adventitial fibroblasts in vitro and in vivo. FOSL1 overexpression significantly increased collagen-related protein expression and induced fibroblast phenotype transformation. However, Naringenin reduced AD incidence and severity in vitro and in vivo. Notably, a decrease in FOSL1 expression enhanced the therapeutic effect of Naringenin in AD. FOSL1 is a key regulator in adventitial remodeling during AD, contributing to the proliferation and fibroblast phenotype transformation of adventitial fibroblasts. FOSL1 could serve as a potential target to improve the sensitivity of Naringenin-based therapy.

Glioblastoma multiforme, a malignant brain tumor has a dismal prognosis and lacks effective treatment. Epidermal growth factor receptor (EGFR) is an attractive drug target for GBM treatment, yet no therapeutic effect has been reported. Here, we have deepened our studies on a recently described EGFR inhibitor, a tetrahydroquinoline-derived triarylmethane, 2-((1,2,3,4-tetrahydroquinolin-8-yl)(4-(trifluoromethyl)phenyl)methyl)phenol (THQPMP), showing a potential cytotoxicity activity against GBM cells LN229 and SNB19. THQPMP exhibits a strong binding affinity to the EGFR receptor of -6.92 kcal/mol, which interacts with 12 amino acid residues. The stable interaction of THQPMP-EGFR complex was validated by molecular simulations dynamics lasting 100 ns. The statistical parameters of partial least squares regression (PLS) revealed robust internal predictive capacity for the Gaussian-based QSAR model developed. The half maximum inhibitory concentration (IC₅₀) for THQPMP and gefitinib was established to be 40.6 µM and 46 µM for LN229 cells and 38.3 µM and 68 µM for SNB19 cells, respectively. SiRNA transfection assay confirmed the specific interaction of THQPMP with EGFR, thus modulating the downstream signaling cascade and inducing cell death in GBM. Furthermore, THQPMP prompted cell cycle arrest at S phase and observed to induce a negligible fold of intracellular calcium level, thereby leading to GBM cell death via a calcium-independent signaling mechanism. A comprehensive ADME analysis was performed, predicting the physicochemical, absorption, distribution, metabolism, and excretion parameters of THQPMP, therefore elucidating its pharmacokinetic and drug likeness properties. THQPMP was validated to cross the blood-brain barrier with moderate permeability. Overall, THQPMP has shown efficient preclinical activity against GBM by modulating EGFR signaling pathways, warranting further in vivo validation for phase clinical trials.