Pharmaceuticals最新文献

{"title":"Folic Acid-Functionalized Chitosan-Coated Oleosomes Engineered for Enhanced Anti-Arthritic Delivery of <i>Syzygium cumini</i> Seed Extract.","authors":"Samar M Aborhyem, Neveen Agamy, Aamena F Abdallah, Shimaa O Ali, Haidy Abbas, Mariam Zewail","doi":"10.3390/ph19101658","DOIUrl":"10.3390/ph19101658","url":null,"abstract":"<p><p><b>Background/Objectives</b>: This study evaluated the therapeutic efficacy of folic acid-functionalized chitosan-coated oleosomes (F-CS-OL) loaded with <i>Syzygium cumini</i> (SYG) seed extract against rheumatoid arthritis (RA) in a complete Freund's adjuvant (CFA)-induced arthritis rat model. <b>Methods</b>: Rats received treatments via the oral route for 11 days following arthritis induction. The colloidal formulation exhibited an average particle size of 80.78 ± 1.02 nm, a zeta potential of -15.5 ± 1.12 mV, and an encapsulation efficiency (EE) of 94.98 ± 0.56%, alongside characterization by TEM, FTIR, and DSC. CFA-induced arthritis produced a pronounced inflammatory and angiogenic response, marked by significantly elevated serum levels of TNF-α, IL-6, CRP, VEGF, ICAM-1, MMP-3, and CXCL13 compared with the normal control group. <b>Results</b>: While treatment with free SYG extract resulted in moderate reductions in these biomarkers, the nanoencapsulated extract (SYG-loaded F-CS-OL) produced significantly greater reductions in inflammatory, chemotactic, matrix-remodeling, and angiogenesis-associated biomarkers compared with the untreated CFA-induced arthritis group. Histopathological examination of joint tissues further supported the biochemical findings. <b>Conclusions</b>: Overall, SYG-loaded F-CS-OL exhibited potent anti-inflammatory, immunomodulatory activities and significantly reduced biomarkers associated with angiogenic signaling in CFA-induced arthritis in CFA-induced arthritis. These findings suggest that folic acid-functionalized nanocarriers may represent a promising strategy for enhancing the therapeutic potential of SYG; however, further pharmacokinetic and safety studies are warranted to support clinical translation.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708674/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting Neuroinflammation After Traumatic Brain Injury: Addressing Sex Differences to Improve Clinical Translation.","authors":"Caitlin Anne Sharratt, Jessica Leigh Hall, Peter John Crack, Hui Xin Tan","doi":"10.3390/ph19101659","DOIUrl":"10.3390/ph19101659","url":null,"abstract":"<p><p>Traumatic brain injury (TBI) is a leading cause of death and disability worldwide. Following the initial mechanical insult, a complex secondary injury cascade is triggered, resulting in neuroinflammation that can exacerbate neuronal damage and contribute to long-term neurological and psychiatric complications. Despite the development of numerous promising therapies targeting neuroinflammation after TBI, none have successfully translated from preclinical research into clinical practice. This persistent translational gap reflects limitations in preclinical and clinical study design, including inadequate representation of the biological heterogeneity of clinical TBI populations. This review critically evaluates the current landscape of therapies modulating neuroinflammatory pathways following TBI, with particular focus on how biological sex can influence therapeutic efficacy. Across multiple preclinical studies, sex-dependent differences were identified in molecular, histological, and functional responses to therapies targeting central regulators of neuroinflammation, indicating that biological sex can influence therapeutic efficacy across multiple inflammatory pathways. The influence of biological sex may also extend beyond preclinical development into clinical trials, where the underrepresentation of females and the failure to incorporate preclinical evidence of sex-dependent pharmacokinetics and therapeutic responses may represent potential contributors to variable or unsuccessful outcomes. Collectively, the evidence reviewed here suggests that biological sex may represent an important, yet underappreciated, contributor to the challenges of therapeutic translation and may obscure clinically meaningful treatment effects in defined patient subgroups. The findings presented throughout this review therefore suggest that biological sex should be considered a core component of therapeutic development, rather than solely as a secondary exploratory variable, to better characterise sex-dependent treatment responses and aid the clinical translation of therapies targeting neuroinflammation after TBI.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708345/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From Design to Synergy: Triazine-Triazole Hybrids as Anti-Breast Cancer Leads Targeting Survival and Invasion Pathways for Augmenting Lapatinib Efficacy.","authors":"Omnia M Ali, Samar Hafez Abbas, Micheal G Shehat, Maged W Helmy, Moustafa H Aly, Mohamed Teleb, Sherine N Khattab, Mohamed Abdel-Aziz","doi":"10.3390/ph19101656","DOIUrl":"10.3390/ph19101656","url":null,"abstract":"<p><p><b>Background/Objectives</b>: Breast cancer is the most common cancer diagnosed and the leading cause of cancer-related deaths among women, ranking fourth overall in both sexes. Its treatment is difficult due to drug resistance and side effects from conventional therapies. Developing multitargeted agents and combination approaches offer a promising way to overcome these challenges. This study details the design and testing of new triazole-triazine hybrids as potential anti-breast cancer drugs, exploring their use in combination with lapatinib to enhance its efficacy in breast cancer. <b>Methods</b>: Sixteen triazine-triazole hybrid compounds with amide and ureido linkers were synthesized via click and coupling protocols and tested for their cytotoxic effects on MCF-7 cells using an MTT assay. The most effective compound was evaluated for its inhibitory effect against AXL using an ADP-Glo™ kinase assay, then examined in combination with lapatinib on cellular and molecular levels. In vitro experiments included chemoinvasion assays for metastasis, flow-cytometric analysis of apoptosis, and ELISA to evaluate the molecular mechanisms of promising hits. Molecular docking studies were conducted to simulate the potential target-hit interactions and predict the plausible structural determinants of activity, paving the way for further lead optimization. <b>Results</b>: Compound <b>6e</b> demonstrated the highest activity (IC₅₀ = 24.58 μM) and good selectivity (SI = 7.87). The combination therapy showed a synergistic effect and increased tumor sensitivity to lapatinib. Biological tests indicated inhibition of the AXL enzyme, downregulation of key pathways (AKT, NF-κB, STAT3, ERK), decreased levels of VEGF, MMP9, and cyclin D1, and increased caspase-3 activity, suggesting apoptosis, which was strengthened by the flow-cytometric analysis findings. Docking results supported multitarget interactions. <b>Conclusions</b>: Compound <b>6e</b> is a promising multitarget anti-breast cancer candidate with synergistic potential in combination with lapatinib, warranting further in vivo studies.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708434/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272286","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative Phytochemical and Biological Evaluation of Walnut Septum and Pellicle Extracts in Chemical, Computational, and Cell-Based Models.","authors":"Letiția Mateș, Denisia Pașca, Ionel Fizeșan, Ana-Maria Vlase, Gabriel Marc, Ionuț-Valentin Creștin, Oana Andreea Alexa, Ioan Tomuță, Lorena Filip, Laurian Vlase, Daniela-Saveta Popa","doi":"10.3390/ph19101657","DOIUrl":"10.3390/ph19101657","url":null,"abstract":"<p><p><b>Background:</b> Walnut (<i>Juglans regia</i> L.) processing generates polyphenol-rich by-products, including the septum (WS) and pellicle (WP). This study comparatively characterized the phytochemical composition of WS and WP from Romania and investigated their antioxidant and preliminary immunomodulatory properties using chemical, computational, and cell-based approaches. <b>Methods:</b> Extracts were characterized by targeted LC-QTOF-MS/MS, and antioxidant capacity was evaluated using ABTS, DPPH, and FRAP assays. Major phenolic compounds were docked against Kelch-like ECH-associated protein 1 (KEAP1), followed by 100 ns molecular dynamics simulations. Cytocompatibility was evaluated in Caco-2, Jurkat, and THP-1 cells, and cellular antioxidant activity was assessed in H₂O₂-stimulated Caco-2 cells using the DCFH-DA assay. Immunomodulatory potential was investigated following lipopolysaccharide (LPS) stimulation of the same cell lines. <b>Results:</b> WSLE contained 25 quantified compounds, with ellagic acid, catechin, procyanidins, and quercitrin among the major quantified constituents, whereas WP contained 13, with higher ellagic acid, gallic acid, and castalagin levels. WPE showed greater cell-free antioxidant capacity, while both extracts attenuated the H₂O₂-induced increase in intracellular DCF fluorescence in Caco-2 cells. Simulations suggested favorable KEAP1 interactions, with quercitrin and catechin predicted to form the most stable complexes. LPS induced an insufficient cytokine response in Caco-2 and Jurkat cells. In THP-1 cells, IL-8 was the only responsive marker, and both extracts reduced its LPS-induced secretion. <b>Conclusions:</b> The extracts showed distinct polyphenolic profiles and redox-related cellular activity. Their effects on IL-8 secretion in THP-1 cells and predicted KEAP1 binding are preliminary and hypothesis-generating; Nrf2 activation was not measured, and validation in physiologically relevant <i>in vivo</i> and clinical models is required to establish the biological relevance and potential applications of these extracts.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708377/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Pathway-Driven Computational Drug Discovery Framework Integrating Machine Learning and Virtual Screening for Identification of Anti-Rheumatoid Arthritis Natural Small Molecules.","authors":"Jijia Sun, Yuezhen Wang, Hong Ye, Jialiang Chen, Yilin Chen, Guangtao Yao","doi":"10.3390/ph19101651","DOIUrl":"10.3390/ph19101651","url":null,"abstract":"&lt;p&gt;&lt;p&gt;&lt;b&gt;Objective:&lt;/b&gt; This study aimed to establish a pathway-driven computational drug discovery framework integrating machine learning, target prioritization, virtual screening, molecular dynamics (MD) simulations, and in vitro evaluation to identify natural small molecules with potential anti-rheumatoid arthritis (RA) activity. &lt;b&gt;Methods:&lt;/b&gt; Sixteen RA-related Gene Expression Omnibus (GEO) datasets were included, with 15 cohorts used for model development and one reserved for independent external validation. Pathway features quantified by gene set variation analysis (GSVA) were subjected to cohort-aware feature selection using a leave-one-dataset-out framework across seven machine-learning algorithms. Predictive modeling and SHapley Additive exPlanations (SHAP) analysis were subsequently performed, followed by random-effects meta-analysis (REMA) and network analysis for candidate-target prioritization. Natural compounds were screened by molecular docking and further evaluated using ligand efficiency, multiple scoring approaches, explicit-membrane MD simulations, and a TNF-α-induced MH7A inflammatory model. &lt;b&gt;Results:&lt;/b&gt; Ten stable RA-associated pathways were identified. The final CAT+ET model achieved ROC-AUCs of 0.783 (95% CI, 0.731-0.836) and 0.610 (95% CI, 0.496-0.720) in the internal and independent external validation sets, respectively, indicating limited cross-cohort generalizability. SHAP and cross-cohort analyses supported Complement and coagulation cascades for downstream investigation, and integrated expression and network evidence prioritized C3AR1 as a candidate target. Virtual screening identified four candidate natural compounds (SM1-SM4). Ligand-size analysis, ligand efficiency, multiple scoring approaches, and explicit-membrane MD simulations provided complementary structural and energetic assessments of these candidates. Among them, SM1 showed comparatively favorable computational characteristics and a relatively consistent reduction in the inflammatory markers IL6 and MMP3 in TNF-α-stimulated MH7A cells, accompanied by changes in &lt;i&gt;C3AR1&lt;/i&gt; expression. These convergent computational and experimental findings supported the prioritization of SM1 for further investigation. &lt;b&gt;Conclusions:&lt;/b&gt; This study established a pathway-driven framework linking cross-cohort pathway identification with candidate-target prioritization and natural-product screening in RA. Complement and coagulation cascades and C3AR1 were prioritized for downstream investigation, while integrated computational and experimental evidence identified SM1 as a candidate with promising anti-inflammatory potential. Although direct compound-C3AR1 engagement and its functional involvement require further validation, the present study provides a systematic and extensible strategy for translating robust RA-associated pathway signals into candidate targets and natural compounds, offering a valuable foundation for subsequent natural-product drug discovery in RA.&lt;/","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708292/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"sDR5-Fc Administered Post-Exposure Alleviates Radiation-Induced Enteric Injury by Inhibiting ROS and NLRP3/Caspase-1/GSDMD-Dependent Pyroptosis and Inflammatory Response.","authors":"Xuan Qiao, Shuang Guo, Zhuang Tian, Lin-Tao Li, Yu-Han Yang, Zhi-Yun Meng, Hui Gan, Shu-Chen Liu, Gui-Fang Dou, Ruo-Lan Gu","doi":"10.3390/ph19101647","DOIUrl":"10.3390/ph19101647","url":null,"abstract":"<p><p><b>Background:</b> The risk of ionizing radiation exposure, including accidents and medical complications, has increased with the rapid development and broad application of nuclear technology. The intestine is highly susceptible to radiation, with radiation-induced intestinal injury characterized by a complex cascade of cellular and molecular reactions. This cascade results in the death of intestinal epithelial cells, which compromises the integrity of the intestinal mucosal barrier and disrupts immune regulation. At present, there are several limitations in the research on the therapy of radiation-induced intestinal injury, including uncertain mechanisms and an absence of optimal pharmaceuticals. This study aims to explore the post-exposure treatment effects of sDR5-Fc (a DR5 antagonist) on radiation-induced intestinal pyroptosis damage. <b>Methods:</b> In this study, C57BL/6 mice, human HIEC-6, and rat IEC-6 intestinal epithelial cells were exposed to a single 15 Gy high-dose γ-irradiation to establish an acute radiation-induced intestinal injury model. All sDR5-Fc treatments were administered after radiation exposure to simulate clinical post-exposure intervention. In vivo histological staining was performed for quantitative evaluation of intestinal damage. In vitro cell viability and LDH release assays were used to evaluate intestinal epithelial injury. Multiple detection methods, including scanning electron microscopy (SEM), flow cytometry, peripheral blood routine analysis, organ index detection, immunohistochemistry, qRT-PCR, and Western blotting, were applied to assess radiation-induced alterations in oxidative stress (ROS, MDA) and pyroptosis-associated inflammasome indicators (NLRP3, caspase-1, GSDMD-N, IL-1β, IL-18). <b>Results:</b> It was observed that treatment with soluble DR5-Fc fusion protein (sDR5-Fc), a DR5 antagonist, markedly reduced these radiation-induced effects. Mice subjected to 15 Gy total abdominal γ-irradiation exhibited similar outcomes after sDR5-Fc administration, including inhibited pyroptosis in intestinal and colonic tissues, enhanced recovery of peripheral blood cells (leukocytes, erythrocytes, and platelets), improved organ indices of the spleen and thymus, and an increased 30-day survival rate. Histological examination and pathological scoring confirmed significant structural damage in the irradiated group, and these pathological alterations were partially reversed following sDR5-Fc administration. Molecular docking and BLI assays suggest a potential in vitro interaction between DR5 and NLRP3, which can be interfered with by sDR5-Fc. Moreover, qRT-PCR and Western blotting assays indicated that sDR5-Fc attenuated pyroptosis by reducing the NLRP3/caspase-1/GSDMD signaling pathway. <b>Conclusions:</b> These findings suggest that sDR5-Fc, as a DR5 antagonist, is a promising therapeutic candidate for mitigating high-dose radiation-induced intestinal injury; however, further research is warranted.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708083/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272465","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryptococcosis After Initiation of Janus Kinase Inhibitors or Biologic DMARDs in Rheumatoid Arthritis: Three Pairwise Active-Comparator Analyses of Propensity Score-Matched TriNetX Cohorts, 2005-2025.","authors":"Ying Xiu Tay, Yi-Ting He, Malcolm Koo, Ming-Chi Lu","doi":"10.3390/ph19101650","DOIUrl":"10.3390/ph19101650","url":null,"abstract":"<p><p><b>Background/Objectives:</b> Comparative safety evidence for cryptococcosis during advanced rheumatoid arthritis (RA) therapy is limited. We compared newly recorded cryptococcosis after initiation of Janus kinase inhibitors (JAKi), tumor necrosis factor (TNF) inhibitors (TNFi), and non-TNF biologic disease-modifying antirheumatic drugs (non-TNF bDMARDs). <b>Methods:</b> Three retrospective active-comparator analyses were conducted in the TriNetX Global Collaborative Network: JAKi versus TNFi, JAKi versus non-TNF bDMARDs, and TNFi versus non-TNF bDMARDs. After 1:1 propensity score matching, outcomes from day 1 to 10 years after initiation, capped at 30 June 2025, were analyzed using Cox proportional hazards models. E-values assessed sensitivity to unmeasured confounding. <b>Results:</b> Matched cohorts included 27,833, 32,440, and 37,351 patients per group, respectively; all absolute standardized mean differences were <0.10. Events occurred in 20 JAKi and 12 TNFi initiators (hazard ratio [HR], 2.20; 95% confidence interval [CI], 1.06-4.54) and in 22 JAKi and 13 non-TNF bDMARD initiators (HR, 2.15; 95% CI, 1.07-4.31). In the secondary comparison, 17 TNFi and 19 non-TNF bDMARD initiators had events (HR, 0.83; 95% CI, 0.43-1.60). Point-estimate E-values for the JAKi comparisons were 3.82 and 3.72; CI-limit E-values were 1.31 and 1.34. <b>Conclusions:</b> JAKi initiation was associated with an approximately twofold higher hazard of newly recorded cryptococcosis than either biologic comparator, while estimated 10-year cumulative event probabilities were below 0.2% in all groups. The separately selected and matched populations do not support an indirect ranking of treatment classes. Low event counts, wide CIs, possible outcome misclassification, fixed exposure classification, and residual confounding preclude causal conclusions. The findings require independent confirmation.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708528/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis, Biological Evaluation, and Molecular Dynamics Studies of Novel 1,2,4-Triazole-Functionalized Benzene-Sulfonate Derivatives as Potential Enzyme Inhibitors and Cytotoxicity in HepG2 Cells.","authors":"Yasemin Ünver, Fikret Türkan, Halil İbrahim Güler, Ercan Oguz, Tuğba Usta, Fatih Çelik","doi":"10.3390/ph19101645","DOIUrl":"10.3390/ph19101645","url":null,"abstract":"<p><p><b>Background/Objectives:</b> In this study, novel benzenesulfonate derivatives containing a 1,2,4-triazole ring were synthesized. <b>Methods:</b> Their molecular structures were characterized by IR, ¹H NMR, and ¹³C NMR spectroscopy, together with Q-TOF LC/MS analyses. For compounds <b>5b</b> and <b>5d</b>, the available mass spectrometric data were not used for structural confirmation, and their structural assignments were based primarily on the spectroscopic data. The inhibitory effects of the synthesized compounds (<b>3</b>-<b>5</b>) against acetylcholinesterase (AChE) and glutathione S-transferase (GST) enzymes were evaluated. <b>Results:</b> Comparison of the results with the reference inhibitors, donepezil and ethacrynic acid, showed that compound <b>3e</b> exhibited the highest AChE inhibitory activity among the tested compounds, with an IC₅₀ value of 1.60 ± 0.198 μM. In addition, compound 4a showed remarkable GST inhibitory activity, with an IC₅₀ value of 1.95 ± 0.492 μM, demonstrating greater potency than the reference inhibitor. The cytotoxic effects of the selected compounds were further assessed against the HepG2 cell line after 24 and 48 h of treatment. Overall, the synthesized molecules exhibited potent enzyme inhibitory activities together with promising cytotoxic effects, highlighting their potential as lead candidates for further biological investigations. In silico analyses were performed to elucidate the molecular basis of the experimentally observed acetylcholinesterase (AChE) inhibitory activity of the synthesized derivatives. Molecular docking against human AChE (PDB ID: 4EY7) identified compound <b>3e</b> as the most favorable binder among the synthesized series, while <b>3f</b> also exhibited a strong binding profile. Subsequent 100 ns molecular dynamics simulations of the hAChE-<b>3e</b>, hAChE-<b>3f</b>, and hAChE-donepezil complexes demonstrated stable protein conformations and persistent ligand accommodation within the catalytic gorge. MM/GBSA calculations further indicated a more favorable binding-energy profile for <b>3e</b> than for <b>3f</b>, whereas donepezil showed the most favorable overall calculated binding energy. Per-residue decomposition highlighted key gorge residues, including Trp86, Trp286, Tyr337, Tyr341, Phe295, and Phe297, as important contributors to ligand stabilization. <b>Conclusions:</b> Collectively, these computational findings provide structural and energetic support for the experimentally observed AChE inhibitory activity of the lead derivatives.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708584/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Practical Roadmap for Implementing the LC-MS-Based Multi-Attribute Method for Analysis of Therapeutic Proteins.","authors":"Jingzhong Guo, Sheila Mugabe, Gantasala Subramanya Sameer Kumar, Vishal Amancha, Li Jing, Niomi Peckham, Jarkko Huuskonen, Dipankar Das, Diane McCarthy","doi":"10.3390/ph19101648","DOIUrl":"10.3390/ph19101648","url":null,"abstract":"<p><p><b>Background/Objectives:</b> The mass spectrometry-based multi-attribute method (MAM) enables site-specific monitoring of multiple product quality attributes (PQAs) within a single LC-MS peptide mapping assay and is increasingly being adopted in biopharmaceutical development and quality control (QC). However, its analytical and operational complexities necessitate a structured approach to method development, qualification, system readiness, and assay transfer. This study presents a practical roadmap for implementing LC-MS-based MAM, aligned with USP-NF General Chapter ⟨1060⟩. <b>Methods:</b> Sample preparation and LC-MS acquisition parameters were optimized to minimize analytical artifacts and support robust workflow performance. System readiness was established using a publicly available monoclonal antibody reference standard. Adalimumab and etanercept were then used as representative biotherapeutics to evaluate key elements of MAM implementation, including qualification and interlaboratory transfer. <b>Results:</b> The refined workflow provided consistent digestion performance, chromatographic stability, mass accuracy, and reliable data processing across both molecules. Qualification studies demonstrated acceptable specificity, precision, sensitivity, and quantitative performance for targeted PQA monitoring, while new peak detection (NPD) assessments supported the detection of emerging or changing species. Interlaboratory transfer studies demonstrated successful transfer of the standardized workflow across laboratories and LC-MS platforms. <b>Conclusions:</b> These results establish a technical and operational foundation for MAM implementation, including system readiness, qualification, and assay transfer. The proposed implementation framework provides a robust and broadly applicable roadmap for MAM deployment across diverse biopharmaceutical programs while supporting subsequent product-specific refinement and validation for routine QC applications.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708689/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272194","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dose-Related Effects of Cannabidiol on Inflammatory and Redox Responses, Bacterial Dissemination, and Multiorgan Injury in CLP-Induced Septic Rats.","authors":"Ufuk Ülker, Bülent Bayraktar, Tünay Karan, Çağrı Çağlar Sinmez, Emre Tüfekçi, Esra Bilici, Umut Alpman, Gökşad Cemil Kotan, Mustafa Yeni","doi":"10.3390/ph19101649","DOIUrl":"10.3390/ph19101649","url":null,"abstract":"<p><p><b>Background:</b> Sepsis couples dysregulated inflammation, oxidative imbalance, microbial dissemination, and multiorgan injury. Cannabidiol (CBD) has anti-inflammatory and redox-modulating properties, but its dose-related effects in polymicrobial sepsis, particularly alongside culture-recoverable bacterial burden, remain incompletely characterized. <b>Methods:</b> Forty-eight male Wistar rats were randomized to Control, Sham, Untreated CLP, or CLP plus CBD at 2.5, 5, or 25 mg/kg (<i>n</i> = 8/group). CBD or vehicle was administered intraperitoneally after cecal ligation and puncture (CLP). Inflammatory, immunometabolic, neurotrophic, oxidative, biochemical, microbiological, and histopathological outcomes were evaluated at 10 h. <b>Results:</b> Untreated CLP increased pro-inflammatory cytokines, asprosin, resistin, oxidative-stress indices, organ-injury biomarkers, and reported culture-recoverable bacterial burdens, while decreasing IL-10, apelin, BDNF, TAS, and immunoreactive SOD, CAT, and GPx concentrations (all omnibus <i>p</i> < 0.001). CBD was associated with progressive improvement across most systemic outcomes, and 25 mg/kg produced the most consistent overall response. Histopathological lesion scores were greatest in Untreated CLP animals and generally lower in CBD-treated groups; however, tissue-level changes were not uniformly dose-ordered across every organ and lesion domain. <b>Conclusions:</b> Within this acute exploratory model, CBD exposure was associated with coordinated improvements across host-response, redox, biochemical, and morphological domains. The microbial summaries suggested lower recoverable burdens, but require animal-level verification of non-detect handling. These findings support pharmacokinetic, safety, mechanistic, survival, and prospectively powered confirmatory studies; they do not establish direct antibacterial activity or a clinically applicable dose.</p>","PeriodicalId":20198,"journal":{"name":"Pharmaceuticals","volume":"19 10","pages":""},"PeriodicalIF":5.7,"publicationDate":"2026-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13708221/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149272342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}