The human metabolic and excretion profile of zalunfiban, a novel glycoprotein IIb/IIIa inhibitor, was studied in a phase 1 clinical trial. C-zalunfiban was administered subcutaneously as a single fixed dose (9.5 mg with 5 μCi total radioactivity).Zalunfiban whole blood concentrations were measured using liquid chromatography-mass spectrometry. C-zalunfiban and metabolites were measured using liquid scintillation counting and accelerator mass spectrometry in whole blood, urine, and feces. Eight participants were enrolled. Zalunfiban was well-tolerated.Following injection zalunfiban was detectable within 5 minutes and the last measurable concentration was observed within 4 hours. The median T was 0.25 hours and mean half-life was 0.96 hours. Zalunfiban accounted for 35.6% of total whole blood radioactivity AUC at 4 hours. The single major identified metabolite was des-gly-zalunfiban (M1), which has <1% of zalunfiban's antiplatelet activity. M1 was the primary urinary metabolite (52.98% of dose) with minor amounts of zalunfiban (1.31%) detected. M1 was also the major metabolite in feces (2.87%) . Total dose recovery reached >90% by 240 hours.Zalunfiban is rapidly metabolized to the nearly inactive M1 metabolite, which is excreted primarily in urine. Renal impairment, therefore, is unlikely to significantly prolong zalunfiban effects and dose adjustment in patients with reduced renal function may not be required.

Osteosarcoma is the most common primary malignant bone tumor in adolescents, with poor prognosis and high rates of recurrence and metastasis. Ferroptosis, a form of regulated cell death associated with iron overload and lipid peroxidation, and mitochondrial dysfunction have been recognized as potential therapeutic targets in cancer. α-Hederin, a natural triterpenoid saponin, has shown antitumor effects in several cancers, but its role in osteosarcoma remains unclear. This study aimed to investigate the pharmacological effects and mechanisms of α-Hederin on osteosarcoma. We integrated network pharmacology and molecular docking to identify potential targets and pathways, and validated the findings using in vitro experiments in 143B osteosarcoma cells. Network pharmacology analysis revealed 27 overlapping targets between α-Hederin and osteosarcoma, with significant enrichment in the PI3K/AKT and ferroptosis pathways. Molecular docking confirmed strong binding affinity between α-Hederin and key targets. Functional assays demonstrated that α-Hederin inhibited cell proliferation and migration, increased intracellular Fe and lipid ROS levels, disrupted mitochondrial membrane potential and ATP production, and downregulated mitochondrial biogenesis proteins. Transmission electron microscopy further revealed typical mitochondrial morphological changes associated with ferroptosis. Western blotting showed decreased expression of GPX4 and SLC7A11, increased ACSL4 levels, and suppressed activation of the PI3K/AKT pathway. Moreover, these effects were partially reversed by the ferroptosis inhibitor Ferrostatin-1. Collectively, these results indicate that α-Hederin exerts antitumor effects against osteosarcoma by inducing ferroptosis and impairing mitochondrial function, partly through inhibition of the PI3K/AKT signaling pathway, providing a potential therapeutic strategy for osteosarcoma.

Phytopharmacology has become a key approach for developing new therapeutic strategies by utilising the diverse bioactive properties of plant-derived compounds to treat complex diseases, including cardiovascular disorders. Myocardial ischaemia-reperfusion (I/R) injury presents a major challenge in the management of acute myocardial infarction by worsening myocardial damage through oxidative stress, apoptosis, and cellular senescence. Carvacrol, a monoterpenoid phenol present in plants such as , possesses potent antioxidant and anti-inflammatory properties. This study investigates carvacrol's cardioprotective effects in an H9C2 cardiac myoblast model of I/R injury.Cardiac myoblast cells were exposed to an ischaemic buffer to simulate I/R conditions, with carvacrol administered at a sub-cytotoxic dose of 12.5 µg/mL prior to exposure. Carvacrol significantly enhanced cell viability by 77.37% restoration, reduced lactate dehydrogenase (LDH) release (from 330.5 ± 25.3 to 160.8 ± 15.7 U/mL,  < 0.01), suppressed reactive oxygen species (ROS) production, inhibited caspase-3 and -8 activities, and mitigated cellular senescence as evidenced by reduced β-galactosidase staining. Additionally, carvacrol restored the expression of the myogenin gene, which was downregulated by ischaemic injury.These findings highlight carvacrol's antioxidant, anti-apoptotic, anti-senescence, and gene-regulatory properties, positioning it as a promising therapeutic candidate for mitigating myocardial I/R injury.

A 5-compartment hybrid model that simulates the pharmacokinetic behavior of methyl mercury [MeHg] and its biotransformation [demethylation] product inorganic mercury [Hg(II)] in humans is described.This model accounts for demethylation of MeHg at two sites; one which is within the body and a second which is extracorporeal (presumably the gut lumen). Simulations indicate that approximately 85% of demethylation occurs in the gut lumen and 15% in body tissues.The 2-compartment model for Hg(II) pharmacokinetics developed by Farris et al. (2008) serves as the basis for the 5-compartment model and is embedded within its structure. Parameters from the 2-compartment model are used to describe Hg(II) pharmacokinetics in the current model.The model is validated against previously published (Smith et al. 1994) experimental data for two human subjects dosed with MeHg.Model simulations showing the effects of decreased biotransformation of MeHg in the gastrointestinal lumen are discussed.

This study evaluated the effects of Serfurosterone A (SSA) from Achyranthes bidentata Bl. on dexamethasone (DEX)-induced osteoblast dysfunction and its intestinal permeability.

This study aimed to apply a machine learning workflow to identify the most relevant biological parameters for predicting both the dose and the route of crack cocaine administration.Seventeen variables were evaluated in rats exposed to different doses of crack cocaine, either intraperitoneally (18 or 36 mg/kg, i.p.) or passive inhalation (25, 50, or 100 mg). Random forest (RF) analysis was used to build predictive models, feature importance analysis to identify key variables, and interaction dependence analysis to explore relationships among variables.Eighty percent of the data was used for training and 20% for testing. The model achieved 85% accuracy in the training phase and 100% in the test phase. During training, the highest accuracy was observed for the 100 mg inhaled group, while the lowest was for the 50 mg inhaled group. Notably, 20% of the 50 mg i.p. cases were misclassified as 36 mg i.p. Feature importance analysis highlighted four key predictors: liver karyolysis, kidney Ki-67 expression, liver binucleation, and escape behaviour.These findings demonstrate that machine learning (ML) can accurately predict both dose and route of crack cocaine exposure, and can highlight biologically relevant parameters involved in the drug's systemic and behavioural effects.

The mutagenicity and bioactivation potential of -nitrosoparoxetine, a -nitrosamine drug substance related impurity (NDSRI) of the marketed antidepressant paroxetine, was characterised .-Nitrosoparoxetine was not mutagenic in an OECD 471-compliant bacterial reverse mutation assay even under experimental conditions that supported oxidative metabolism by CYP enzymes. The major biotransformation pathways of -nitrosoparoxetine paralleled the ones previously noted with paroxetine. Virtually all stable metabolites of -nitrosoparoxetine in human, rat, and hamster liver tissue were derived from phase 2 conjugations on an unstable catechol intermediate, generated from the oxidative 1,3-benzodioxole ring scission by CYP enzymes. The piperidine ring in -nitrosoparoxetine was resistant to the α-carbon oxidation step leading to piperidine ring scission, and to the eventual formation of a DNA-reactive electrophilic species.CYP reaction phenotyping studies demonstrated that paroxetine was exclusively metabolised by human CYP2D6, whereas -nitrosoparoxetine was subject to metabolism by multiple human CYP isoforms including CYP2C19, CYP2D6, and CYP3A4.Whether most NDSRIs derived from parent amine precursors that are resistant to α-carbon oxidation by CYPs will also be devoid of a mutagenic response needs to be scrutinised further with additional examples to bolster structure-genotoxicity relationships of complex NDSRIs.

Xevinapant, a drug previously under development for head and neck cancer, was investigated to determine its intrinsic clearance (CL) and assess the involvement of CYP enzymes in its metabolism. Extensive studies showed very low turnover, necessitating advanced methodologies to accurately measure CL values.Two novel in vitro assays, using a modified suspension hepatocyte protocol or 3D hepatocyte spheroid cultures, allowed measuring hepatic turnover of xevinapant with very low CL values of 1.3 or 0.3 µL/min/10 cells, respectively. Reaction phenotyping using specific inhibitors, enabled by enhanced detection capabilities in spheroids, identified primarily CYP3A (> 50%) as contributor to the hepatic metabolism of xevinapant.- extrapolation, in combination with data from the human mass balance study, revealed that hepatic metabolism, primarily mediated by CYP3A, accounts for approximately 30% of the overall clearance. The remaining clearance is renal (∼20%) and metabolic, but potentially extrahepatic (∼50%).This research highlights the importance and benefits of utilising advanced techniques, such as human hepatocyte spheroids, to accurately characterise drug metabolism, providing valuable insights for pharmacokinetic evaluations and, ultimately, supporting the development of a PBPK model and understanding the DDI risk of xevinapant.

Silver diamine fluoride (SDF) is commonly used in clinical settings to manage dental caries in young children. A major drawback of SDF is its tendency to darken carious lesions, turning them brown or black, which has drawn criticism. This study investigates the cytotoxic effects of SDF on human gingival fibroblasts (hGF).hGF cells were exposed to varying concentrations of SDF (0.0001%, 0.001%, and 0.005%) for a 24-hour period. We analysed cytotoxicity, the potential for reactive oxygen species (ROS) production, mitochondrial function, morphological indicators of apoptosis, and key molecular markers of apoptosis.The findings revealed that SDF exposure led to significant, dose-dependent cytotoxicity and ROS generation in hGF. Morphological changes consistent with apoptosis were observed. Furthermore, SDF disrupted the balance of Bax and Bcl-2 proteins and reduced the expression of proliferating cell nuclear antigen in these cells. In animal studies, single SDF application for 3 h resulted in mild structural alterations of gingival tissue and basilar hyperplasia in rats.Overall, the findings demonstrate that SDF exhibits cytotoxicity towards hGF, primarily through intracellular ROS production and subsequent apoptosis. Additionally, the experiments validate the mild toxic effects of SDF on rat gingival tissues. Developing controlled drug delivery approaches could help mitigate SDF's toxicity in clinical applications.

Aflatoxin B (AFB) is a potent hepatocarcinogen. It is activated by cytochrome P450 3A4 (CYP3A4) into a DNA-reactive epoxide linked to liver cancer.Six scopoletin metabolites were predicted using SmartCYP. Their lipophilicity was assessed via BioTransformer 3.0; molecular docking was performed with CBDock 2 using CYP3A4 (PDB ID: 1TQN).Phase I Metabolite 2 showed the highest binding affinity (Vina score: -7.4 kcal/mol) and was selected for a 100 ns molecular dynamics (MD) simulation using Desmond with the OPLS3e force field.The metabolite maintained stable binding (RMSD < 1.0 Å), showed >80% hydrophobic interaction occupancy with Phe215, Phe108, and Leu210, and formed transient hydrogen bonds with Ser119; MM-GBSA predicted a binding free energy of -75 kcal/mol.Scopoletin metabolites may competitively inhibit AFB activation by binding CYP3A4, indicating potential for chemoprevention pending experimental confirmation.

This investigation was planned to evaluate the mechanistic interactions of Imidacloprid (IM) and Chlorothalonil (CL), inducing endocrine disruptions after 28 days of oral administration.Male rats were divided into a control (CR) and three treatment groups, receiving IM (45 mg/kg), CL (300 mg/kg), and an admixture of IM and CL.The gathered findings indicate that IM and CL induced spuriously abnormal thyroid hormone measurements distinguished by a decrement in circulating levels of T3, T4, with a normal/slight increase in TSH concentration. Equally, a sharp decline in insulin levels was recorded in the IM (0.46 ± 0.33 pmol/L) and CL (2.08 ± 0.43 Pmol/L) groups compared to the healthy specimens (8.21 ± 0.014 Pmol/L). Additionally, high plasma alanine transferase (ALT/SGPT) and aspartate transferase (AST/SGOT) levels, as well as alkaline phosphatase (Alk-P) and metabolite concentrations, including serum creatinine (SCR), uric acid (SUA), and urea levels, were recorded in all treated groups. Both IM and CL significantly compromised the antioxidant defense system by increasing the MDA level, decreasing the GSH content, and inhibiting the activity of CAT and SOD.We systematically investigated the binding affinity and the potential inhibitory effects of IM and CL on a series of enzymes integral to the blood transport and receptor binding of THs, such as MCT8 and TSHR. A significant binding affinity of IM and CL to these proteins was observed, suggesting the possibility of competitive or allosteric inhibition. The MD simulations revealed the strong and stable interactions between IM, CL, MCT8, and TSH-R. MMGBSA energies showed that both pesticides compete with hormones at active sites, indicating their potential to modulate key enzymes involved in thyroid hormone transport and action.Therefore, it is anticipated that this result supply beneficial knowledge for future therapeutic endeavors.

Black American Ginseng (Panax quinquefolium L.) is a traditional Chinese medicine taken orally. While saponins from American Ginseng are known to enhance memory, the effects and mechanisms of Black American Ginseng saponins remain unclear.This study investigates their potential in alleviating memory impairment in ageing mice. UPLC-Q-Orbitrap-MS/MS was used to identify compounds in Black American Ginseng, and network pharmacology predicted potential targets. The Morris water maze test assessed cognitive function, while Western blot and ELISA measured BDNF, TrkB, inflammatory markers (IL-1β, TNF-α, IL-6), and the activation of the PI3K/AKT/CREB signalling pathway. Networking pharmacology and PPI analysis identified PIK3CA, EGFR, and PIK3R1 as key targets, with KEGG enrichment highlighting the PI3K/AKT pathway. Behavioural tests confirmed that Black American Ginseng saponins significantly improved memory in ageing mice. Molecular analyses revealed upregulation of BDNF and TrkB and suppression of IL-1β and IL-6. Additionally, Western blot confirmed activation of the PI3K/AKT/CREB pathway, supporting its neuroprotective role.Black American Ginseng saponins enhance cognitive function by modulating neurotrophic signalling and reducing neuroinflammation.These findings provide new insights into their potential therapeutic application for age-related cognitive decline.

Considering the important role of mitochondria in mammalian cells and the close correlation between mitochondrial dysfunction and various diseases, this study evaluated the potential toxicity of glyphosate on mitochondrial function in swine heart mitochondria.The results indicated that glyphosate did not have a significant effect on mitochondrial respiration, mitochondrial swelling, and FF-ATPase activity under the experimental conditions tested.These findings suggest that, in this model, glyphosate alone may not exert a direct cardiotoxic effect on mitochondrial bioenergetics.Caution is warranted when extrapolating these results to human health, as differences in species, exposure levels, and experimental conditions may influence outcomes.The toxicity observed in glyphosate-based herbicides could involve adjuvants or synergistic interactions between glyphosate and other formulation components, as proposed by other studies.

This study aimed to develop a targeted metabolomics workflow using low-resolution tandem mass spectrometry (MS/MS) to identify metabolic alterations in zebrafish () embryos exposed to environmentally relevant concentrations of the antibiotic's amoxicillin and clarithromycin.Zebrafish embryos were exposed to the lowest concentrations (1 µg/L) and the highest concentration (1 mg/L) of amoxicillin and clarithromycin. A library-assisted multiple reaction monitoring-enhanced product ion (MRM-EPI) approach was applied using a QTRAP LC-MS/MS system, enabling the detection and structural confirmation of 108 endogenous metabolites. A targeted MRM-EPI method was subsequently optimised for sensitivity, reproducibility, and specificity. Multivariate statistical analyses, including principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), were performed to assess metabolic differences between exposed and control groups.Exposure to both antibiotics resulted in significant alterations in amino acid, purine, lipid, and energy metabolism, indicating that even sub-lethal concentrations can disrupt vital physiological processes in zebrafish embryos. These findings highlight the sensitivity of metabolomics for detecting early biochemical perturbations and support the use of zebrafish embryos as a practical and ethically suitable model for environmental toxicity assessment. The developed MRM-driven workflow provides a reproducible platform for predictive toxicology and ecological risk evaluation.

Norovirus is a leading cause of acute gastroenteritis worldwide, yet no approved antivirals currently exist. In this study, we employed approaches to evaluate apigenin derivatives as potential inhibitors of norovirus RNA-dependent RNA polymerase (RdRp). Seventy-three compounds were initially retrieved from the ZINC database, of which 36 satisfied Lipinski's Rule of Five and were advanced for detailed analysis. Pharmacokinetic predictions revealed high intestinal absorption and favorable drug-likeness profiles for most derivatives, with limited central nervous system penetration, desirable for targeting gastrointestinal infections. Molecular docking identified ZINC14636470 (A1) as the top candidate with strong binding affinity (-9.9 kcal/mol) and key hydrogen bond interactions at the RdRp active site. Molecular dynamics simulations confirmed the stability of the A1-RdRp complex (RMSD = 0.20 ± 0.03 nm; Rg = 2.39 ± 0.02 nm). While AMES predictions flagged some derivatives as potentially mutagenic, scaffold optimization may mitigate these risks. Apigenin derivatives, particularly A1, demonstrate promising inhibitory potential against norovirus RdRp, combining favorable pharmacokinetic properties with stable enzyme binding. These findings provide a computational foundation for subsequent and validation, supporting the development of natural flavonoid-based scaffolds as antiviral leads against norovirus.

Adipose-derived stem cells (ADSCs) offer a novel approach for treating orthopaedic diseases. Flurbiprofen is a non-steroidal anti-inflammatory analgesic drug used in bone injury therapy. However, its cytotoxicity to ADSCs remains unclear.The differentiation potential of ADSCs was analysed using the differentiation induction method. Microscopic analysis, Live/Dead staining, MTT assay, and CCK8 assay were used to determine the cytotoxic effects of flurbiprofen on ADSCs at different concentration gradients. Vascular endothelial growth factor (VEGF) levels were measured by ELISA to reflect flurbiprofen's effects on the activity and paracrine function of ADSCs. ANOVA was applied, with  < 0.05 considered significant and  < 0.01 highly significant.Live cell concentration decreased dose-dependently with flurbiprofen treatment, particularly on the 7th day. The level of VEGF decreased significantly with the increase of the concentration of flurbiprofen in a certain concentration range, which indicated that flurbiprofen could inhibit the activity and secreted proteins of ADSCs.Our study showed that small doses of flurbiprofen did not affect the secretion of proteins by cells and cell activity. Therefore, it is necessary to pay attention to the concentration of flurbiprofen in the clinical application of ADSCs therapy for orthopaedic diseases.

1. To study the effects of calycosin on palmitic acid-induced HepG2 cells, as well as the potential mechanisms of action. 2. Potential targets of calycosin for the alleviation of insulin resistance were predicted by network pharmacology. Glucose concentration in the culture medium was determined by the GOD-POD method. The model of insulin resistance was established by palmitic acid-induced HepG2 cells. Effects of palmitic acid and calycosin on HepG2 cell activity were determined using an MTT assay kit. The expression levels of AKT1 and FOXO3a were detected by western blot. The expression level of hsa-miR-324-3p was detected by RT-qPCR. Dual luciferase reporter assay to detect targeting of AKT1 by hsa-miR-324-3p. 3. AKT1 was predicted and validated as a potential target of calycosin for treatment of insulin resistance. The model of insulin resistance was successfully established by palmitic acid-induced HepG2 cells. Up-regulation of AKT1 expression inhibits FOXO3a entry into the nucleus. Calycosian was demonstrated to concentration-dependently increase the sensitivity of insulin resistance cells to insulin. The hsa-miR-324-3p was proven to exist in insulin-resistant cells. Hsa-miR-324-3p was found to target AKT1 involved in the alleviation of insulin resistance. 4. Calycosin inhibits FOXO3a nuclear translocation by regulating the hsa-miR-324-3p/AKT pathway, thus alleviating insulin resistance.

This study aimed to investigate the therapeutic potential of Aloe-emodin (AE) for irritable bowel syndrome (IBS), focusing on its effects and underlying mechanisms.Deoxycholic acid (DCA)-induced IBS rats (Sprague-Dawley) were orally administered AE. Body weight and faecal pellet were monitored. Anxiety-like behaviour, visceral hypersensitivity, colon permeability were assessed the open-field (OF) test, abdominal Withdrawal Reflex (AWR) score, FITC-dextran fluorescence, respectively. Enzyme-linked immunosorbent assay (ELISA) quantified substance P (SP), 5-hydroxytryptamine (5-HT), TNF-α, and IL-6 levels. Hypoxia inducible factor-1α (HIF-1α) expression was analysed qRT-PCR. The mechanism of AE on IBS was evaluated in LPS-treated NCM460 injury.AE alleviated IBS symptoms (improved weight gain, reduced faecal output/water content, increased centre exploration time, lowered AWR scores, decreased colon permeability, SP, 5-HT, and pro-inflammatory cytokine levels). HIF-1α upregulation in colonic tissues and LPS-induced NCM460 cells was suppressed by AE treatment. The protective effect of AE was reversed by HIF-1α overexpression in IBS rats. AE enhanced cell proliferation, reduced cellular permeability, and inflammation in LPS-stimulated NCM460 cells. HIF-1α overexpression partially reversed the protective effects of AE in LPS-induced NCM460 injury.AE ameliorated IBS symptoms by promoting cell proliferation, suppressing cell permeability, and inflammation of colonic epithelial cells regulating HIF-1α.

Astragalin (AST), a flavonoid, shows promise for neurodegenerative diseases like Parkinson's disease (PD), cognitive impairment (CI), and depression. However, its efficacy in treating neurodegenerative diseases and the underlying molecular mechanisms remain unclear.This study aims to evaluate the metabolite profile, pharmacokinetics, toxicity, molecular targets, and potential biological activities of AST. Thirty-one AST metabolites formed through Phase II reactions (O-glucuronidation, O-sulfation, and methylation) were found.AST and its metabolites partially violate Lipinski's Rule of Five, including molecular weight and hydrogen bond donors, impacting drug-likeness. However, AST and its metabolites have favourable safety and potential anti-neurodegenerative and antidepressant effects.AST shows strong binding affinities with key neuroinflammatory targets, including IL1B, IL6, TNF, NOS2, PTGS2, SERT, caspase-3, caspase-8, and GABAa receptor, and network analysis highlights its association with neuroinflammatory pathways.Collectively, these findings support AST as a potential neurotherapeutic candidate and offer a basis for further and validation.

It has been shown that fluoxetine is cytotoxic on pancreatic beta-cells induction of mitochondrial dysfunction and oxidative stress. We investigated the direct effect of fluoxetine on isolated pancreatic mitochondria and evaluate the potential protective effects of betanin and thymoquinone.Mitochondria were isolated from rat pancreas and treated with various concentrations of fluoxetine (10-8000 µM). Then, protective effect of betanin (100-500 µM) and thymoquinone (10-100 µM) on fluoxetine-induced mitochondrial toxicity were studied (60 min). The activity of succinate dehydrogenases (SDH), reactive oxygen species (ROS) formation, mitochondrial swelling, mitochondrial membrane potential (MMP) collapse, malondialdehyde (MDA) production and glutathione level were analysed.Fluoxetine directly caused toxicity in pancreatic isolated mitochondria at concentration of 500 μM and higher. Except MDA and GSH, fluoxetine caused significantly SDH activity reduction, MMP collapse, mitochondrial swelling and ROS formation in pancreatic mitochondria. However, our results showed that only betanin protected fluoxetine-induced mitochondrial dysfunction, while thymoquinone had no impact on mitochondrial toxicity induced by fluoxetine.We can conclude that fluoxetine is directly toxic on pancreas isolated mitochondria, which may be related to its diabetogenic potential in humans. Moreover, our finding suggested that use of betanin may be beneficial for prevention of diabetogenic effect of fluoxetine.

Permethrin, a pyrethroid commonly used to grow crops worldwide, may affect the aquatic environment and show toxicological impact on non-target organisms.Farmed shrimp, , can be included among these organisms since enzymatic antioxidant defense responses were disrupted in gills and hepatopancreas after exposure to this xenobiotic.Phase II biotransformation (Glutathione S-Transferase - GST activity) was affected as well as Acetylcholinesterase - AchE activity.Phase II biotransformation (Glutathione S-Transferase - GST activity) was affected as well as Acetylcholinesterase - AchE activity.1.