Introduction This study aims to systematically elucidate the specific targets and molecular mechanisms by which modified Chaihu Shugan San (mCSS) inhibits hepatocellular carcinoma (HCC) progression to link potential targets with cell cycle and immune microenvironment. Methods mCSS targets were first predicted via TCM databases, followed by functional enrichment analysis and drug-target-pathway network construction. TCGA HCC transcriptomic data underwent differential expression analysis and clinical screening to obtain prognosis-related differentially expressed genes. Core herbal formula-modulated targets were identified by intersecting predicted drug targets with these prognostic genes. Public HCC single-cell RNA-seq data was analyzed to characterize single-cell expression patterns of core targets for mechanism exploration. Finally, the pharmacological effects were validated via serum pharmacology assays in HCC cell lines. Results Network pharmacology analysis identified 319 potential targets of mCSS for HCC treatment. Transcriptomic analysis of TCGA data revealed 353 differentially expressed prognosis‑related genes. Intersection analysis uncovered 11 core targets: SPP1, AKR1C3, MMP9, PPIA, XDH, EPHX2, CDK4, GAPDH, HSP90AA1, LGALS1, and ADAM9. Single-cell data analysis demonstrated that CDK4 expression was markedly elevated in hepatoma cells (R=0.226，P < 0.001) and fibroblasts (R=0.117，P < 0.001), whereas LGALS1 expression was significantly increased in fibroblasts (R=0.426，P < 0.001) and T cells (R=0.348，P < 0.001). Cell-cell communication analysis indicated enhanced fibroblast-mediated communication and collagen-related signaling in HCC. In Hepa1–6 cells, mCSS-containing serum inhibited cell proliferation (P < 0.05) and migration and reduced CDK4 protein expression and Rb phosphorylation. Moreover, scTenifoldKnk-based virtual knockout of CDK4 revealed perturbations in complement/coagulation cascades, integrin signaling, and ECM–receptor interactions, suggesting that CDK4-related proliferative and microenvironment-associated networks may contribute to the observed effects of mCSS. Conclusion These findings support a multi-target anti-HCC effect of mCSS involving CDK4-associated signaling, whereas LGALS1-related stromal remodeling remains a predicted mechanism requiring further experimental validation.
Background BCL-XL is a key anti-apoptotic protein frequently overexpressed in various malignancies, where it contributes to therapy resistance. PROTAC technology offers a potential strategy to expand the therapeutic window by selectively degrading BCL-XL in tumor cells while sparing platelets. Herein, we present selected insights into the research and development of a novel BCL-XL PROTAC. Objectives This study aimed to characterize a CRBN-recruiting BCL-XL PROTAC series and identify key preclinical optimization considerations for BCL-XL degradation. Methods CRBN-based BCL-XL PROTACs were synthesized with medicinal chemistry methods and evaluated using MOLT-4 and RS4;11 cell viability assays, liver microsomal stability assays, mouse pharmacokinetic studies, BCL-XL degradation assays, platelet toxicity assays, and a MOLT-4 T-ALL xenograft model. Results A series of CRBN-based PROTACs achieved low-nanomolar cellular potency compared with the benchmark degrader DT2216. 25 exhibited potent BCL-XL degradation activity and reduced platelet toxicity. However, PK analysis revealed higher systemic clearance and lower plasma exposure. In a MOLT-4 xenograft model, 25 was well-tolerated but demonstrated limited tumor growth suppression. Conclusions This work established a proof-of-concept for CRBN-recruiting BCL-XL PROTACs as a platelet-sparing therapeutic strategy. Although 25 demonstrated superior in vitro potency, its in vivo antitumor efficacy was constrained by its suboptimal pharmacokinetic profile. This study serves as an exploratory framework for future optimization programs.
Objective To systematically evaluate the efficacy of tripterygium glycosides combined with immunosuppressants or glucocorticoids in the treatment of various types of connective tissue disease-interstitial lung disease (CTD-ILD). Methods Databases including PubMed, Web of Science, Cochrane Library, CNKI, WanFang, and VIP were searched to collect literature on the efficacy of tripterygium glycosides in the treatment of CTD-ILD published up to December 2025. Extracted data included basic information of the study, patients’ general information, treatment plans, and outcome indicators. Statistical analysis was performed using R 4.4 software, and publication bias was assessed. Results A total of 10 studies were included, involving 749 cases (363 in the experimental group and 386 in the control group). The control group primarily received treatment with glucocorticoids or immunosuppressants, including prednisone, methylprednisolone, methotrexate, sulfasalazine, cyclophosphamide, and leflunomide. The experimental group received treatment with tripterygium glycosides combined with glucocorticoids or immunosuppressants. Meta-analysis showed that the total effective rate was higher in the experimental group than in the control group (RR=1.16, 95%CI=1.09–1.24, Z = 4.47, P < 0.001). Subgroup analysis by disease type revealed no significant difference in total efficacy rates between RA-ILD (RR=1.21, 95%CI=1.04–1.42) and CTD-ILD (RR=1.29, 95%CI=1.13–1.47) (P = 0.07). Subgroup analysis by treatment regimen indicated significant differences in total efficacy rates between tripterygium glycosides plus immunosuppressants (RR=1.21, 95%CI=0.99–1.48) and tripterygium glycosides plus glucocorticoids (RR=1.28, 95%CI=1.15–1.44) (P = 0.03). Subgroup analysis by dosage showed no significant difference in total efficacy rates between tripterygium glycosides at 60 mg/day (RR=1.14, 95%CI=1.07–1.22) and tripterygium glycosides at 1.5 mg/(kg·day) (RR=1.47, 95%CI=1.13–1.90) (P = 0.07). Subgroup analysis by treatment duration revealed significant differences in total efficacy rates between tripterygium glycosides at a 6-month course (RR=1.30, 95%CI=1.19–1.43) and tripterygium glycosides at a 3-month course (RR=1.05, 95%CI=0.96–1.15) (P = 0.001). No significant difference was observed in the incidence of adverse reactions between the two groups (RR=0.82, 95%CI=0.50–1.35, Z = 0.77, P = 0.44). The funnel plot suggested no publication bias. Conclusion Tripterygium glycosides exhibit favorable efficacy in treating CTD-ILD, and no increased risk of adverse reactions has been observed. However, due to the lack of key safety data on reproductive toxicity and impairment of liver and kidney function, its long-term safety requires further evaluation. It is particularly suitable for patients with strong dependence on glucocorticoids or those requiring long-term immunosuppressants but showing poor therapeutic outcomes. It is recommended to administer at a standard dosage of 60 mg/day, with a treatment course lasting no less than 6 months.
Background Lung cancer remains the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer (NSCLC) representing a major subtype characterized by high incidence and mortality. Despite advances in targeted therapy and immunotherapy, treatment resistance and the immunosuppressive tumor microenvironment remain major clinical challenges. The combination of herpes simplex virus thymidine kinase and interleukin-12 (HSV-TK/IL-12) has demonstrated anti-tumor potential, but its role and mechanism of action in NSCLC remain poorly defined. This study aims to systematically evaluate the anti-tumor efficacy of HSV-TK/IL-12 combination gene therapy in NSCLC and to investigate the potential involvement of Connexin 43 (Cx43) in the bystander effect (BSE) alongside immune activation. Methods Overexpression vectors for HSV-TK and IL-12 were constructed to study their functions in NSCLC. Protein expression was measured by western blot. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry were employed to assess cell survival and apoptosis, respectively. In vitro, human peripheral blood lymphocytes were used to evaluate immune responses relevant to clinical immunotherapy. In vivo, xenograft mouse models were used to evaluate effects on tumor growth and experimental lung colonization. Results Overexpression of HSV-TK or IL-12 significantly increased their respective protein levels in A549-TK and H1299-TK cells (P < 0.01). Ganciclovir (GCV) treatment reduced the viability of A549 and H1299 cells in a dose-dependent manner (P < 0.05). Importantly, the combination therapy of HSV-TK and IL-12 induced apoptosis in A549 and H1299 cells (P < 0.05). In vitro, the combination also enhanced the proliferation and cytotoxic activity of co-cultured human peripheral blood lymphocytes (P < 0.05). A pronounced BSE was observed in the presence of HSV-TK/GCV, and its magnitude was positively correlated with the proportion of TK + cells and with Cx43 expression levels (P < 0.05). In vivo, the combination of HSV-TK and IL-12 inhibited tumor growth and reduced lung nodule formation in a tail-vein colonization model (P < 0.05). Conclusion The combination of suicide gene HSV-TK and immunomodulatory gene IL-12 exerts a stronger anti-tumor effect than either monotherapy in NSCLC, demonstrating significant potential for clinical translation as a novel gene therapy strategy for NSCLC patients, particularly those with treatment-resistant disease.
Aim of the study To explore the protective mechanism of Xuebijing against myocardial injury caused by organophosphorus poisoning, providing a theoretical basis for its prevention and treatment. Materials and methods An organophosphorus poisoning model was established by intraperitoneal injection of dichlorvos (O, O-dimethyl-O-2,2-dichlorovinyl phosphate, DDVP). Xuebijing or atropine was administered immediately after DDVP treatment. Echocardiography, biochemical assays, and HE staining were used to evaluate myocardial injury. MitoSOX fluorescent probe, JC-1 fluorescent probe, and ELISA were utilized to assess mitochondrial oxidative stress. Prussian blue staining, iron content detection kit, and western blot assay were applied to assess ferroptosis. Bioinformatics analysis indicated that Xuebijing alleviates the ferroptosis-related gene HNRNPD in myocardial injury caused by organophosphorus poisoning and the ferroptosis-related gene PDK4. In vitro experiments confirmed that Xuebijing mediates the HNRNPD/PDK4 signaling pathway to modulate mitochondrial oxidative stress-induced ferroptosis and improve organophosphorus poisoning-induced myocardial injury. Results DDVP induced myocardial injury, ferroptosis, and mitochondrial oxidative stress in rats. Inhibiting ferroptosis improved DDVP-induced myocardial injury, and inhibiting ROS reversed the promoting effect of DDVP on cardiomyocyte ferroptosis. Xuebijing inhibited DDVP-induced myocardial injury, ferroptosis, and mitochondrial oxidative stress. Mechanistically, Xuebijing-induced HNRNPD alleviated DDVP-induced cardiomyocyte ferroptosis by binding to and stabilizing PDK4 mRNA, upregulating its expression, restraining pyruvate dehydrogenase activity, and reducing mitochondrial ROS production. Conclusion Xuebijing ameliorates organophosphorus poisoning-induced myocardial injury by upregulating the HNRNPD/PDK4 signaling pathway, inhibiting mitochondrial oxidative stress, and attenuating ferroptosis.
Background sepsis-induced cardiomyopathy (SIC) is a critical complication of sepsis, contributing significantly to high mortality rates. The underlying molecular mechanisms remain poorly understood, and effective therapeutic strategies are urgently needed. Xuebijing (XBJ), a traditional Chinese medicine formulation, has shown potential in treating sepsis due to its anti-inflammatory and antioxidant properties. However, its specific role in regulating SIC and the key molecular targets involved have not been fully elucidated. Methods Differentially expressed genes (DEGs) between septic patients and healthy controls were identified using the GEO dataset (Series accession: GSE95233). Weighted gene co-expression network analysis (WGCNA) was employed to determine sepsis-related gene modules. Machine learning algorithms (LASSO regression, SVM-RFE, and Random Forest) were used to screen key therapeutic targets of XBJ. Functional enrichment (Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Gene set enrichment analysis) were performed to explore biological pathways. The relative abundance of immune cell types in normal and disease samples was estimated using the CIBERSORT algorithm. Molecular docking validated the binding affinity between XBJ components and key targets. In vitro, H9C2 cells were treated with lipopolysaccharide (LPS) to simulate sepsis-induced cardiac injury, and the effects of XBJ and PADI4 overexpression on inflammation and oxidative stress were assessed by enzyme-linked immunosorbent assays, colorimetric assays, and fluorometric assay. In vivo, a cecal ligation and puncture (CLP) rat model was used to evaluate XBJ’s therapeutic efficacy. Results WGCNA identified the “MEblue” module as the most clinically significant in sepsis (r = -0.81, P = 2e-29). By intersecting XBJ component targets, DEGs, and module genes, 20 candidate genes were identified. Machine learning algorithms further narrowed these to three key genes: N-ribosyldihydronicotinamide:quinone dehydrogenase 2 (NQO2), peptidyl arginine deiminase 4 (PADI4), and protein kinase C eta (PRKCH). PADI4 showed the most significant differential expression in sepsis and strong binding affinity with XBJ components (binding energy < −6 kcal/mol). Immune infiltration analysis revealed significant alterations in immune cell populations in sepsis (P < 0.05), with PADI4 expression correlating with neutrophil degranulation and T-cell differentiation pathways. In vitro, XBJ attenuated LPS-induced inflammation and oxidative stress in H9C2 cells (P < 0.05), but these effects were reversed by PADI4 overexpression (P < 0.05). In vivo, XBJ improved cardiac function (elevated ejection fraction and fractional shortening) and reduced inflammation and oxidative stress in CLP-induced sepsis rats (P < 0.05). Conclusion XBJ ameliorated sepsis-induced cardiomyopathy by inhibiting PADI4 to suppress inflammation and oxidative stress. These findings highlight PADI4 as a potential biomarker and therapeutic target for SIC.
Background Refractory acute gout in elderly patients with multimorbidity is difficult to manage because standard anti-inflammatory therapies, including nonsteroidal anti-inflammatory drugs, colchicine, and glucocorticoids, may be ineffective or contraindicated. Interleukin-1β (IL-1β) inhibition represents a potential alternative for high-risk patients with limited treatment options. Methods We report the clinical course of a 69-year-old male with a 40-year history of refractory gout complicated by hypertension, type 2 diabetes mellitus, valvular heart disease, hepatic dysfunction, bilateral multiple renal cysts, and recent maxillofacial infection. The patient presented with severe polyarticular inflammation and markedly elevated high-sensitivity C-reactive protein (hs-CRP, 152.4 mg/L), with poor response to prior oral diclofenac. Because conventional anti-inflammatory therapies were unsuitable, a single 200 mg subcutaneous dose of the IL-1β inhibitor firsekibart was administered. Clinical symptoms, visual analogue scale (VAS) pain score, inflammatory markers, serum urate, organ function, recurrence, and adverse events were monitored during treatment and four months of follow-up. Results After firsekibart administration, pain and joint inflammation improved rapidly. The VAS score decreased from 7 at baseline to 2 by day 7, and hs-CRP normalized to 3.9 mg/L by day 6. During the four-month follow-up, the patient remained free of gout flares and reported no adverse events. Conclusion This case suggests that firsekibart may provide rapid and pronounced anti-inflammatory effects in elderly patients with refractory gout and complex multimorbidity, supporting the use of IL-1β inhibitors in high-risk populations with limited treatment alternatives. Further studies with larger samples and longer follow-up are needed to confirm its safety and effectiveness in high-risk real-world populations.
Background Alzheimer’s disease (AD) affects a large number of people worldwide; its pathological mechanisms are complex, and there is a lack of effective treatments. HM568, a honokiol derivative, has been shown in previous studies to have therapeutic potential for Parkinson’s disease, but its efficacy and mechanisms of action in other neurodegenerative diseases remain unclear. Objective This study aims to investigate the effects and mechanisms of HM568 on Alzheimer's disease through reverse virtual screening and experimental validation. Methods Reverse virtual screening using AutoDock Vina was performed to identify potential targets of HM568. KEGG, Gene Ontology (GO), and protein–protein interaction (PPI) analyses, together with molecular dynamics simulations, were conducted to explore the associated mechanisms. A D-galactose/AlCl₃-induced AD mouse model and an Aβ₂₅–₃₅-injured PC12 cell model were established. Behavioral tests, histopathological staining, biochemical assays, ELISA, cellular functional analyses, Western blotting, and RT-qPCR were used to evaluate the effects of HM568. Results Reverse virtual screening identified voltage-dependent anion channel 1 (VDAC1) as a key AD-related target of HM568, showing strong binding (−9.83 kcal/mol) and stable interactions during 100 ns simulations. In vivo, HM568 improved the overall condition and cognition of AD mice. Body weight gain was increased by approximately 72% relative to the model group, and Morris water maze performance was markedly improved (P < 0.05–0.001). HM568 also reduced hippocampal neuronal damage, increased antioxidant enzyme activities, and decreased brain levels of inflammatory cytokines, Aβ₁₋₄₂, and AChE activity (all P < 0.05). In vitro, HM568 dose-dependently inhibited Aβ₂₅–₃₅ aggregation and reduced apoptosis of Aβ₂₅–₃₅-injured PC12 cells from 36.13% to 12.56% at 4 μM (P < 0.001). Furthermore, HM568 significantly suppressed ROS accumulation, restored mitochondrial membrane potential, increased ATP production, and upregulated mitochondrial respiratory chain complexes I and IV. These protective effects were accompanied by reduced VDAC1 expression and modulation of apoptosis-related genes, including decreased Cytc, Bax, Caspase-3, and Caspase-9 and increased Bcl-2. Conclusion HM568 exerts a neuroprotective effect by inhibiting the abnormal overexpression of VDAC1 and improving mitochondrial function, thereby suppressing neuronal apoptosis. This establishes an experimental foundation for developing and applying honokiol derivatives in neurodegenerative disease treatment while providing novel insights and directions for future anti-AD drug development.
Background Nanotechnology has transformed modern medicine by addressing critical challenges in drug delivery, diagnostics, and biomaterials engineering. The integration of nanoscale materials into pharmaceutical formulations has enabled the development of advanced therapeutic platforms with improved targeting, controlled drug release, enhanced bioavailability, and reduced systemic toxicity. Concurrent advances in nanotheranostics, biomimetic systems, and bio-inspired materials have expanded the clinical potential of nanotechnology for precision medicine and regenerative healthcare. Methods This review critically evaluates recent advances in nanotechnology-enabled pharmaceuticals and biomaterials by synthesizing evidence from peer-reviewed literature. The review covers major classes of nanocarriers, including lipid-based nanoparticles, polymeric nanoparticles, dendrimers, micelles, and inorganic nanomaterials, together with their synthesis, functionalization, characterization, and biomedical applications. Emerging developments in stimuli-responsive drug delivery, nanotheranostics, biomimetic nanomaterials, tissue engineering, artificial intelligence-assisted nanomedicine, personalized medicine, and 3D bioprinting are also discussed, with emphasis on translational and clinical perspectives. Results Recent innovations demonstrate that nano-enabled drug delivery systems significantly improve therapeutic efficacy through site-specific targeting, controlled and sustained drug release, enhanced pharmacokinetics, and reduced adverse effects. Multifunctional nanotheranostic platforms enable simultaneous diagnosis and therapy, facilitating early disease detection and personalized treatment. Biomimetic and bio-inspired nanomaterials exhibit enhanced biocompatibility, immune modulation, and tissue regeneration, while the integration of artificial intelligence and advanced manufacturing technologies has accelerated nanocarrier design, optimization, and clinical translation. Conclusion Nanotechnology continues to redefine pharmaceutical sciences and biomaterials research by enabling safer, more effective, and patient-centered therapeutic strategies. Despite challenges related to large-scale manufacturing, regulatory approval, long-term safety, and clinical translation, continued interdisciplinary research is expected to accelerate the development of next-generation nanomedicines and functional biomaterials, supporting the advancement of precision medicine and improving global healthcare outcomes.
Background Idarucizumab, andexanet alfa, and four-factor prothrombin complex concentrate (4F-PCC) are the principal strategies for anticoagulant reversal. In December 2025, andexanet alfa was withdrawn from the U.S. market because of post-marketing thrombosis concerns. This withdrawal renewed scrutiny of the post-marketing safety profiles of reversal agents. Objective To systematically characterize and compare adverse-event (AE) signal profiles for these three agents in the U.S. FDA Adverse Event Reporting System (FAERS). Methods FAERS quarterly ASCII files were standardized and de-duplicated according to FDA rules. Reports from 2013Q2 to 2026Q1 that listed idarucizumab, andexanet alfa, or 4F-PCC as primary or secondary suspect drugs were analyzed. Disproportionality at the system-organ-class (SOC) and preferred-term (PT) levels was assessed with four algorithms. Additional analyses included target-event clustering, andexanet alfa versus 4F-PCC head-to-head (H2H) comparison, Weibull time-to-onset modeling, temporal trends, mortality regression, and sensitivity analyses. Results A total of 3950 AE reports were identified: 1558 for idarucizumab, 1050 for andexanet alfa, and 1342 for 4F-PCC. Strong PT signals numbered 39, 29, and 32, respectively. Andexanet alfa showed the most prominent thromboembolic profile, with thromboembolic events in 37.8% of reports (ROR 23.54) and cerebral infarction as the leading signal (ROR 146.24). It was also the only agent with a strong nervous-system SOC signal and showed early-onset thrombosis in the Weibull analysis (median 1 day, β 0.66). The andexanet thrombotic ROR increased across the pre-advisory, post-advisory, and post-withdrawal periods, paralleling the andexanet alfa versus 4F-PCC H2H ROR. After full adjustment, mortality remained highest for idarucizumab, with lower odds for andexanet alfa and 4F-PCC overall (OR 0.52 and 0.18) and in the intracranial hemorrhage (ICH) subgroup (0.30 and 0.25). Conclusion The three reversal agents showed distinct AE profiles. Andexanet alfa showed persistent, early-onset thromboembolic signals through the advisory and withdrawal periods. Idarucizumab showed hemorrhagic and critical-illness signals, with residual differences that may reflect unmeasured ICH severity. These findings support individualized risk management and proactive post-marketing surveillance.
Background The functional profile of 4-thioxo chromen-2-one analogues, including antimicrobial efficacy, serum protein binding, and photophysical properties, remains largely unexplored despite the pharmacological relevance of the coumarin scaffold. Objectives This study aimed to provide an integrated characterization of derivatives 1a–g by evaluating antibacterial activity against Gram-positive pathogens, investigating BSA binding interactions via biophysical and computational methods, and assessing intrinsic fluorescence properties. Methods Antibacterial activity against Staphylococcus aureus (PTCC-1112) and Bacillus subtilis (PTCC-1015) was determined by broth microdilution to ascertain MIC and MBC following 24 h incubation at 37 °C. BSA conformational changes were probed using Far-UV circular dichroism (CD) spectroscopy (195–260 nm), with secondary structure fractions deconvoluted by CDNN. Molecular docking simulations employed AutoDock 4.2 (PyRx 0.8) using BSA (PDB: 4F5S). Intrinsic fluorescence was characterized in DMSO (λₑₓ = 360 nm). All experiments were performed in triplicate. Statistical analyses comprised one-way and two-way repeated-measures ANOVA with Dunnett's and Tukey's HSD post-hoc tests. Results Compound 1d exhibited potent activity against S. aureus (MIC = 0.625 μg/mL; MBC = 2.5 μg/mL), while 1g demonstrated exceptional potency against B. subtilis (MIC = MBC = 0.312 μg/mL). SAR revealed pathogen-dependent substituent preferences. Compound 1c was inactive against both strains (MIC > 10 μg/mL) and exhibited the weakest BSA binding affinity among the tested derivatives (ΔG = -7.5 kcal/mol).CD spectroscopy showed that 1 f induced the greatest reduction in BSA α-helical content (Δfα = −0.110). Docking confirmed spontaneous binding at subdomain IIA (ΔG = −7.5 to −9.3 kcal/mol), with 1 f showing the strongest affinity (-9.3 kcal/mol). Notably, no significant correlation was observed between antibacterial potency and BSA affinity; the most potent antimicrobial, 1g, displayed comparable binding (ΔG = -8.9 kcal/mol) to 1e (ΔG = -8.7 kcal/mol), while the inactive compound 1c exhibited the weakest binding. This observation suggests that BSA binding affinity is not a primary determinant of antimicrobial efficacy in this series. All compounds exhibited substituent-dependent intrinsic fluorescence. Conclusions The 4-thioxo chromen-2-one scaffold represents a promising multifunctional platform with lead compounds 1d and 1g showing potent, pathogen-selective antibacterial activity. The dissociation between antimicrobial efficacy and serum protein binding suggests structural features for target engagement differ from those favouring high-affinity BSA occupancy, a pharmacologically favourable profile. Intrinsic fluorescence provides a potential handle for future bioimaging applications. These findings establish a strategic foundation for lead optimization; however, mechanistic elucidation and in vivo pharmacokinetic studies are required.
Background Curcuma longa (turmeric), Zingiber officinale (ginger), and Allium sativum (garlic) are widely used in traditional medicine, often in combination, for the management of febrile illnesses, including malaria. However, the scientific basis for their combined efficacy and underlying mechanisms remains insufficiently understood. This study investigated the phytochemical composition, molecular interactions, and antiplasmodial activity of a polyherbal formulation comprising these medicinal plants. Methods The ethanol extract of the polyherbal formulation was analysed using Gas Chromatography–Mass Spectrometry (GC–MS) to identify major phytoconstituents. Molecular docking, pharmacophore modelling, molecular dynamics simulations, and Molecular Mechanics/Poisson–Boltzmann Surface Area (MM/PBSA) analyses were employed to evaluate interactions with the Plasmodium falciparum chloroquine resistance transporter (PfCRT). In vivo antiplasmodial activity was assessed in Plasmodium berghei-infected mice, alongside haematological, biochemical, and oxidative stress parameters. Acute oral toxicity was also evaluated. Results GC–MS analysis identified 21 bioactive compounds, with oleic acid (15.84%), cyclopropane derivatives (9.24%), 2-buten-1-one (8.96%), squalene (8.43%), ar-turmerone (6.24%), α-zingiberene (6.21%), phytol (5.52%), and Cholest-14-en-3-ol (4.76%) as major constituents. Molecular docking revealed that Cholest-14-en-3-ol and dehydroabietol exhibited the strongest binding affinities toward PfCRT (−8.4 kcal/mol), exceeding that of chloroquine (−5.4 kcal/mol). Molecular dynamics simulations confirmed the stability of the Cholest-14-en-3-ol–PfCRT complex over 100 ns, while MM/PBSA analysis yielded a favourable binding free energy of −33.13 kcal/mol. In vivo, the polyherbal formulation achieved 97% suppression of parasitaemia by day 4, comparable to chloroquine (98%) and significantly greater than that of the untreated infected group (P < 0.05). Treatment also significantly improved haematological indices, reduced serum aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase activities, increased superoxide dismutase and glutathione peroxidase activities, and decreased malondialdehyde levels compared with infected controls (P < 0.05). No signs of acute toxicity were observed at doses up to 5000 mg/kg body weight. Conclusions The polyherbal formulation demonstrated potent antiplasmodial activity supported by both computational and in vivo evidence. The observed effects may be mediated through favourable interactions with PfCRT, suppression of parasite growth, restoration of haematological and hepatic function, and attenuation of oxidative stress. These findings provide scientific support for the traditional use of this polyherbal combination and warrant further mechanistic and clinical investigations.
Background In recent years, various Chinese medicines have been widely applied in the treatment of rheumatoid arthritis (RA). This study integrated network pharmacology, machine learning, and cell experiments, aiming to explore the action mechanism of Xiaobi Pill in alleviating RA. Objectives To identify key targets of Xiaobi Pill in RA and validate the underlying mechanism Methods Network pharmacology analysis was performed through screening of active ingredients from Xiaobi Pill, online target prediction, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO) enrichment analysis. Candidate targets were subsequently screened by machine learning (LASSO and RF algorithms). Molecular docking was completed using AutoDock Vina. Tumor necrosis factor-alpha (TNF-α) was used for in vitro RA induction in human fibroblast-like synoviocytes (HFLS). Cell viability was detected by Cell Counting Kit-8 (CCK-8) assay. The protein expression was measured using Western blotting. Inflammatory cytokines were examined using Enzyme-Linked Immunosorbent Assays (ELISA). Cell apoptosis was assessed via Terminal-deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Results This study predicted 518 targets from ingredients of herbs in Xiaobi Pill, and 506 RA-associated targets. A total of 77 targets were common to Xiaobi Pill and RA. KEGG and GO enrichment analysis results showed the associated pathways and functional orientation of these 77 genes. Machine learning screened matrix metallopeptidase 13 (MMP13) and spleen tyrosine kinase (SYK) as two candidate targets, and molecular docking indicated the interactions between some ingredients and MMP13 or SYK protein. Cell experiments indicated that Neoastilbin relieved TNF-α-induced inflammation, enhanced apoptosis, and suppressed the nuclear factor kappa-B (NF-κB) pathway in HFLS cells. These regulatory effects of Neoastilbin on HFLS cells were attributed to MMP13 downregulation and subsequent NF-κB signaling inhibition. Conclusion MMP13 was screened as a key target of Neoastilbin (a component of Xiaobi Pill) for RA alleviation through network pharmacology and machine learning. Cell validation suggested that Neoastilbin mitigated inflammatory response and promoted apoptosis of HFLS in an in vitro RA model via targeting the MMP13/NF-κB pathway.
Background Atherosclerosis is a leading cause of cardiovascular morbidity and mortality. It is initiated by endothelial injury and progresses through persistent oxidative stress, endothelial senescence, and macrophage foam cell formation. Although current therapies primarily target lipid metabolism and inflammation, effective strategies that catalytically eliminate reactive oxygen species (ROS) and alleviate vascular senescence remain limited. Objective This study aimed to develop biocompatible copper indium selenium nanozymes (CIS NPs) with antioxidant enzyme-mimicking activities and evaluate their therapeutic potential against oxidative stress, cellular senescence, and atherosclerosis progression. Methods CIS NPs were synthesized and comprehensively characterized for morphology, crystallinity, elemental composition, and colloidal stability. Their superoxide dismutase (SOD)- and glutathione peroxidase (GPx)-like catalytic activities were evaluated in cell-free systems. The antioxidant and anti-senescent effects of CIS NPs were investigated in hydrogen peroxide-induced human umbilical vein endothelial cells (HUVECs). Their effects on foam cell formation were assessed in RAW 264.7 macrophages using Dil-oxLDL uptake and Oil Red O staining. Therapeutic efficacy and biosafety were further evaluated in high-fat diet-fed ApoE-/- mice. Results CIS NPs exhibited dual SOD- and GPx-like catalytic activities, enabling sequential scavenging of superoxide anion (O2•−) and hydrogen peroxide (H2O2). In HUVECs, CIS NPs significantly reduced intracellular ROS levels, attenuated oxidative injury, and alleviated cellular senescence, as evidenced by decreased SA-β-gal staining. In RAW 264.7 macrophages, CIS NPs reduced Dil-oxLDL uptake and lipid accumulation, thereby suppressing foam cell formation. In ApoE-/- mice, repeated intravenous administration of CIS NPs markedly decreased atherosclerotic plaque burden, vascular ROS accumulation, and senescent cell abundance, without obvious abnormalities in the preliminary histological, biochemical, and hemolysis assessments. Conclusions CIS NPs function as efficient antioxidant nanozymes that mitigate oxidative stress, endothelial senescence, and macrophage foam cell formation. By targeting multiple pathological processes involved in atherosclerosis, CIS NPs effectively attenuate disease progression in vivo and represent a promising nanotherapeutic strategy for the prevention and treatment of atherosclerosis.
Background Colorectal cancer (CRC) remains one of the most prevalent malignancies worldwide with high mortality, largely due to local recurrence and distant metastasis. Cysteinyl leukotriene receptor 1 (CysLTR1) is aberrantly overexpressed in CRC and associated with poor prognosis. Zafirlukast, a selective CysLTR1 antagonist approved for asthma treatment, has demonstrated anti-tumor effects in several cancers, yet its role and mechanism in CRC remain unexplored. This study aimed to elucidate whether zafirlukast exerts anti-tumor effects in CRC and to investigate the underlying molecular mechanism. Methods Cell Counting Kit-8 (CCK-8), wound healing, Transwell, and flow cytometric assays were used to test the cellular phenotypes. The relationship between CysLTR1 and phosphoinositide-3-kinase adapter protein 1 (PIK3AP1) was predicted using the MEM database. RT-qPCR and Western blotting detected the expression of CysLTR1, PIK3AP1, apoptotic proteins, and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway-related proteins. Results In this study, zafirlukast dose-dependently inhibited CRC cell proliferation, migration, and invasion, but promoted apoptosis (P < 0.01). Zafirlukast reduced CysLTR1 and PIK3AP1 expression and inactivated PI3K/AKT signaling (P < 0.001). CysLTR1 was correlated with PIK3AP1, and CysLTR1 overexpression elevated PIK3AP1 expression (P < 0.001). PIK3AP1 upregulation or the AKT activator SC79 partially reversed the anti-tumor effects of Zafirlukast in CRC cells (P < 0.001). Conclusions Zafirlukast suppressed PIK3AP1 and inactivated the PI3K/AKT signaling pathway via in a manner involving CysLTR1, contributing to the inhibition of CRC cell proliferation, migration, and invasion.
Background This study aimed to evaluate the potential of transferosome-based nanocarriers for enhancing brain delivery of desvenlafaxine (DSV). Improving central nervous system uptake of antidepressants remains clinically significant for optimizing therapeutic efficacy and diagnostic imaging applications. Methods A series of DSV-loaded transferosome formulations were prepared using surfactants such as Tween 80, Solutol H15, and D-α-tocopheryl polyethylene glycol succinate (TPGs) as edge activators. Formulations were characterized by particle size, encapsulation efficiency, and zeta potential. In vivo biodistribution and pharmacokinetic studies were conducted in mice following intravenous administration of 131I-DSV solution or 131I-DSV-loaded transferosomes. Results Transferosome formulations achieved encapsulation efficiencies of 54%, 63%, and 70%, with particle sizes of 89.06, 204.1, and 75.3 nm and zeta potentials of −49.6, −55.8, and −56.38 mV, respectively. All formulations significantly increased brain uptake compared with the free drug solution. The TPGS-based formulation demonstrated superior performance, achieving the highest encapsulation efficiency and enhanced brain delivery. Conclusions Surfactant-based transferosomes, particularly TPGs-containing systems, markedly improved brain uptake of desvenlafaxine compared with free drug. These findings support the potential of radiolabeled transferosomes as promising nanocarriers for brain-targeted delivery and suggest their applicability in neurological disorder imaging.
ANKRD30A (Ankyrin Repeat Domain 30 A), also known as NY-BR-1, is a tissue-restricted gene initially identified as a breast cancer-associated antigen and later classified as a cancer-testis antigen due to its selective expression in breast, prostate, and testicular tissues. Emerging evidence suggests that ANKRD30A plays context-dependent roles in cancer biology, immune regulation, and reproductive processes. This review provides a comprehensive synthesis of current knowledge regarding the molecular characteristics, expression dynamics, immunological relevance, and translational significance of ANKRD30A across multiple disease contexts. Differential expression of ANKRD30A has been reported in hormone-responsive cancers such as estrogen receptor-positive breast cancer and prostate cancer, whereas reduced expression is observed in aggressive malignancies, including triple-negative breast cancer and subsets of lung cancer. These expression patterns indicate potential utility as a diagnostic and prognostic biomarker. Recent transcriptomic and multi-omics studies also associate ANKRD30A with immune-cell infiltration, modulation of the tumour microenvironment, hypoxia-related pathways, and responses to immune checkpoint therapy, although most evidence remains correlative. Beyond oncology, genome-wide association studies implicate ANKRD30A in autoimmune disorders and behavioural traits, while emerging reproductive studies suggest possible involvement in spermatogenesis and fertility. Despite growing interest, the mechanistic understanding of ANKRD30A remains limited due to insufficient functional validation and a lack of standardised experimental models. This review highlights critical knowledge gaps and emphasises the need for integrated multi-omics analyses, functional genomics, spatial transcriptomics, and translational studies to clarify the biological and clinical significance of ANKRD30A in precision medicine, reproductive and cancer genetics.
Background Anthracene and chalcone-based derivatives possess diverse pharmacological properties; however, comprehensive studies integrating their synthesis, structural characterization, quantum chemical investigation, molecular docking, molecular dynamics (MD) simulation, ADMET evaluation, and experimental anticancer assessment targeting Carbonic Anhydrase II (CA-II) remain limited. This study aimed to synthesize and comprehensively evaluate a novel anthracene–chalcone derivative, (E)-1-(Anthracen-9-yl)-3-(4-chlorophenyl)prop-2-en-1-one (PCPO), as a potential CA-II-targeted anticancer lead compound. Methods PCPO was synthesized via Claisen–Schmidt condensation with a 90% yield and characterized using FT-IR and NMR spectroscopy. Density functional theory (DFT) calculations, including geometry optimization, natural bond orbital (NBO), frontier molecular orbital (FMO), molecular electrostatic potential (MEP), Mulliken charge, intrinsic reaction coordinate (IRC), UV–Vis, electronic circular dichroism (ECD), and density of states (DOS) analyses, were performed to investigate its structural and electronic properties. Molecular docking against CA-II (PDB ID: 6VJ3) was carried out, followed by a 100 ns molecular dynamics simulation to evaluate complex stability. Drug-likeness and pharmacokinetic properties were predicted using ADMET analysis. The in vitro anticancer activity was assessed by MTT assay, and the IC₅₀ value was compared with that of doxorubicin. Results DFT calculations confirmed a planar and highly conjugated molecular structure with a CO bond length of 1.258 Å. NBO analysis revealed significant π→π*, σ→σ*, and n→π* interactions contributing to electronic stabilization, while FMO analysis showed an energy gap of 5.4081 eV, indicating moderate chemical reactivity. MEP and Mulliken charge analyses identified O27 as the most electron-rich region and Cl38 as the electron-deficient center. The IRC analysis confirmed a single transition state connecting reactants and products, and DOS analysis suggested semiconducting behavior with an approximate band gap of 0.8 eV. Molecular docking demonstrated a binding affinity of −9.0 kcal/mol, which was stronger than that of the native ligand (−7.5 kcal/mol) and tamoxifen (−7.2 kcal/mol), supported by hydrogen bonding with GLN92 and hydrophobic interactions involving VAL135, PRO202, and LEU198. The 100 ns MD simulation demonstrated a stable protein–ligand complex with low RMSD and RMSF fluctuations throughout the simulation. ADMET prediction indicated acceptable drug-like characteristics, including a molecular weight of 342.08 Da and favorable membrane permeability, although high lipophilicity (logP = 6.028) and limited aqueous solubility were observed. MTT cytotoxicity evaluation against A549 human lung cancer cells demonstrated concentration-dependent antiproliferative activity, with an IC₅₀ value of 12.6 µg/mL, comparable to that of doxorubicin (13.8 µg/mL). Conclusions The integrated experimental and computational analyses demonstrate that PCPO possesses favorable structural stability, electronic characteristics, strong CA-II binding affinity, stable protein–ligand interactions, and promising in vitro antiproliferative activity. These findings support PCPO as a promising lead scaffold for the further development and optimization of Carbonic Anhydrase II-targeted anticancer agents, while additional in vivo and mechanistic studies are required to validate its therapeutic potential.
A new imidazole-based heterocycle, 4-(2-(2-(4-isopropylphenyl)-4,5-diphenyl-1H-imidazol-1-yl)ethyl)morpholine, was synthesized via a one-pot multicomponent reaction and its structure was confirmed by FT-IR, ¹H NMR, and ¹ ³C NMR spectroscopy. The vibrational characteristics associated with aromatic and aliphatic functionalities exhibited excellent agreement between the experimental and DFT-predicted IR spectra at the B3LYP/6–311 ++G(d,p) level. The GIAO-calculated NMR chemical shifts closely matched the experimental values, confirming the electronic environment around the heteroaromatic core. Geometry optimization revealed a rigid π-conjugated imidazole ring system connected to a flexible morpholine-ethylene chain with energetically favorable conformers at −180° and +60°. NBO analysis demonstrated strong π→π* and n→π* delocalization, contributing to enhanced electronic stabilization. FMO and MEP analyses indicated a favorable charge distribution for receptor binding, while the calculated first-order hyperpolarizability suggests that the compound possesses moderate nonlinear optical (NLO) properties. The synthesized compound also exhibited significant in vitro cytotoxic activity against A549 human lung cancer cells in the MTT assay, indicating its potential as an anticancer agent. Molecular docking against PI3Kα showed a high binding affinity (−9.8 kcal/mol) with diverse interactions, including π–cation, π–anion, hydrogen bonding, and hydrophobic contacts. Furthermore, 200 ns molecular dynamics simulations confirmed the stability of the protein–ligand complex (RMSD 2–3.5 Å). ADMET predictions indicated favorable drug-like characteristics; however, potential toxicity suggests that further structural optimization is warranted. Overall, the excellent agreement between experimental and theoretical studies, together with the promising cytotoxic, computational, and physicochemical properties, highlights this multifunctional imidazole derivative as a promising lead for anticancer drug development and advanced material application.
Background Indoleamine and Tryptophan 2,3-dioxygenase (IDO/TDO) are crucial immunosuppressive enzymes in cancer immunotherapy. This study aimed to systematically benchmark the predictive utility of diverse molecular descriptors, encompassing classical fingerprints and modern deep learning embeddings, for selectivity prediction of IDO/TDO inhibitors. Methods A dataset comprising 760 compounds was utilized for multi-class classification. The investigation evaluated a comprehensive array of molecular representations, including Morgan fingerprints (both bit- and count-based), MACCS keys, Mol2vec, ChemDist, and ChemBERTa embeddings. Classification models included k-Nearest Neighbors (kNN) and Support Vector Machine (SVM), tested both with and without internal and external class balancing techniques such as SMOTE and ADASYN. Model performance was assessed using both random and scaffold-based splitting protocols. Results SVM demonstrated superior classification scores compared to kNN in random data splitting. Internal class weighting outperformed external resampling techniques (SMOTE/ADASYN) regarding minority class F1-scores. The best model, SVM trained on the 2048 count-based Morgan fingerprints (AUC-ROC = 0.88, MCC = 0.65), achieved superior performance on the internal test set. Interestingly, kNN models were shown to be more predictive than SVM applied on the Bemis-Murcko scaffold split data. Accordingly, kNN model applied on 300-dimensional Mol2vec embeddings showed competitive results (AUC-ROC = 0.77, MCC = 0.51) compared to the kNN applied on the 1024-bit Morgan fingerprints (AUC-ROC = 0.77, MCC = 0.53). Conclusion Established circular fingerprints remain an effective and reliable representation for selectivity prediction of IDO/TDO inhibitors compared to modern neural network embeddings. The obtained results are context-dependent and cannot be generalized to other classes of molecules.
