Archives of Toxicology最新文献

Kinase inhibitors (KIs) are an increasing class of drugs targeting protein kinase activity in various pathologies, but predominantly in cancer and autoimmune diseases. Despite successful clinical applications, one of the challenges that remains is their safety, since many KIs lack selectivity and inhibit multi kinases or block pathways that are also important for homeostasis of healthy cells. This may lead to adverse reactions in several critical target organs of toxicity, including heart, liver and kidney. Inflammatory cytokine-mediated intracellular signaling is critical for the control of cell survival and adverse outcome and are modulated by diverse kinases. Likewise, inhibition of kinase activities by on- or off-target effects of KIs may contribute to adverse effects. In this study we systematically screened 760 different KIs targeting the majority of the kinome for their ability to affect TNFα-mediated expression of ICAM1 in hiPSC-derived renal proximal tubule-like cells (PTLCs), cardiomyocytes-like cells (CMLC) and hepatocyte-like cells (HLC). Palbociclib (CDK4/6), miltefosine (PI3K/AKT/mTOR), gilteritinib (FLT3/AXL), and erdafitinib (FGFR) led to increased expression of the ICAM1-eGFP reporter and the release of chemokines. Kinase inhibitor activity data from the ChEMBL database indicated off-target kinase inhibitor activity associated with enhanced ICAM1-eGFP expression. Finally, clinically relevant KIs that enhanced ICAM1-eGFP expression in PTLCs were found to be associated with renal adverse events in patients. Our overall findings support the application of imaging-based hiPSC-derived ICAM1-eGFP reporter covering different differentiated cell lineages to identify liabilities of novel kinase inhibitors to modulate TNFα pathways.

Chronic cadmium exposure, even in environmental doses, has been linked to multiple metabolic disturbances, including white adipose tissue (WAT) dysfunction. WAT dysfunction is defined as a loss of endocrine, immunologic, and metabolic homeostasis, characterized by a low-grade, progressive, and non-resolving inflammation development, namely adiposopathy. This study evaluated the immunometabolic effects of Cd exposure in drinking water on WAT of male Wistar rats, using concentrations of 15 and 32 ppm (environmental doses) over periods of up to 5 months. Inflammatory markers in serum and tissue were analyzed, along with macrophage phenotype, NF-κB pathway activation, leptin and adiponectin expression, correlations with the adiponectin/leptin (A/L) index, and the development of fibrosis. The results showed a progressive increase in proinflammatory cytokines (IL-6, TNF-α, IL-1β), sustained NF-κB activation, and a shift from anti-inflammatory (CD206⁺) to proinflammatory (CD16⁺) macrophages. These changes were accompanied by dysregulation of the adiponectin/leptin axis and a decrease in the A/L ratio, with dynamic correlations to immune markers. Fibrosis was detected in late stages. In conclusion, our results demonstrated for the first time that Cd exposure in environmental doses induces adiposopathy; thereby, findings indicate that Cd can progressively disrupt the immunometabolic homeostasis of adipose tissue, promoting an inflammatory and profibrotic environment with potential implications for the development of metabolic diseases.

Nitazenes have recently surfaced the illicit opioid market, causing numerous intoxications and fatalities. N-Pyrrolidino derivatives, protonitazepyne and metonitazepyne, have circulated since 2023 and have been involved in overdose intoxications. Their pharmacological properties remain largely unknown. However, pharmacokinetic/dynamic data are crucial for clinicians and toxicologists to manage intoxications and interpret legal cases. Protonitazepyne and metonitazepyne metabolism was assessed using human hepatocyte incubations and blood/urine from an intoxication case; samples were analyzed with liquid chromatography-high-resolution mass spectrometry and software-aided data mining. µ-(MOR), κ-(KOR), and δ-(DOR) opioid receptor activation was assessed using a GTP G_i binding assay. MOR docking was simulated with UCSF Chimera and AutoDockSuite. Pharmacological relevance of major metabolites was predicted through in silico MOR docking. Major metabolites were produced through nitroreduction, pyrrolidine N-dealkylation, and oxidation to N-butanoic acid and O-dealkylation. Protonitazepyne and metonitazepyne potencies at MOR were 3.7 and 11.5 nmol L^-1, respectively; efficacies were 154 and 101%. Partial agonism and low potency were observed at KOR/DOR. In silico inhibition constants at MOR for protonitazepyne, 5-amino-protonitazepyne, metonitazepyne, and 5-amino-metonitazepyne were 0.68, 11.45, 1.98, and 2,050 nmol L^-1, respectively. Protonitazepyne and metonitazepyne are MOR-selective full agonists, with potencies about seven and two times higher than fentanyl. These nitazenes present significant health risks through central nervous system/respiratory depression. Their primary metabolites showed lower/marginal in silico MOR affinity, suggesting they might be pharmacologically active, albeit to a much lesser extent than the parent compounds. We propose 5-amino derivatives (blood) and N-butanoic acid derivatives (urine) as biomarkers for detecting consumption.

Ovatoxin-a (OVTX-a) is the main palytoxin (PLTX) analogue produced by the marine benthic dinoflagellate Ostreopsis cf. ovata from the Mediterranean area, where it poses risks for human health mainly through inhalation of marine aerosols and/or skin contact with contaminated seawater. Herein, we highlight the effects of OVTX-a, in comparison to PLTX, at the skin level using human HaCaT keratinocytes, employing a sample containing OVTX-a at 86% purity. After 4 h exposure, OVTX-a reduced cell viability (EC₅₀ = 8.3 × 10^-9 M) with one order of magnitude lower potency than PLTX (EC₅₀ = 3.7 × 10^-10 M). Accordingly, OVTX-a induced lower levels of cell necrosis and mitochondrial depolarization than PLTX, but a 1-h-exposure to each toxin provoked similar increases in ROS production. Moreover, cells co-exposure to ouabain, an inhibitor of Na⁺/K⁺ ATPase, the molecular target of PLTX, or to diphenyliodonium chloride, a non-specific inhibitor of flavoprotein-based enzymes involved in PLTX-induced oxidative stress, suggested that OVTX-a and PLTX share not only the same molecular target, but also the same mechanism of ROS-dependent cytotoxicity. Overall, these results provide new evidence on OVTX-a hazard characterization at the skin level, clarifying its mechanism of action at the basis of cutaneous adverse effects experienced by bathers during O. cf. ovata blooms.

Fentanyl and its analog valerylfentanyl (CAS: 122882-90-0) represent a major toxicological threat amid the ongoing opioid crisis. Using an integrative in silico approach (STopTox, ProTox 3.0, TEST 5.1.2, VEGA QSAR, Percepta, ADMETlab, admetSAR), we evaluated key toxicity end points relevant to clinical and forensic toxicology. Valerylfentanyl exhibited high acute toxicity with predicted LD₅₀ values as low as 18.0 mg/kg (ProTox) and 150.13 mg/kg (TEST). Both compounds showed strong potential to inhibit the hERG potassium channel (95.7% for valerylfentanyl), indicating a substantial risk of cardiotoxicity. Predicted organ-specific effects were most prominent in the lungs (94%), cardiovascular (89%), and gastrointestinal systems (81%), with moderate impact on kidneys and blood. Skin irritation potential was high (up to 81.6%), while genotoxicity remained low, though valerylfentanyl showed non-negligible mutagenic probability in some models. The structural analysis identified piperidine-related toxicophores responsible for these effects. Our findings confirm that minor structural modifications can significantly alter toxicological profiles. These results underscore the application of integrated in silico approach for rapid, ethical hazard identification of emerging NPS, supporting their critical application in both clinical and forensic toxicology.

Heated tobacco products (HTPs) have emerged as alternatives to conventional cigarettes, with claims of reduced health risks. However, this notion remains controversial due to limited evidence, necessitating further investigation of their toxicological effects. This study examined the cytotoxic effects of HTP aerosols in parallel with cigarette smoke. Whole cigarette smoke condensates (WCSCs) were prepared from three commercially available HTPs (Lil, iQOS, and Glo) and 3R4F reference cigarettes, capturing both gas and particulate phases. Cytotoxicity was assessed in the human bronchial epithelial cell line BEAS-2B treated with 200-1200 µg/mL HTP-WCSCs or 40-240 µg/mL 3R4F-WCSC using four distinct assays including lactate dehydrogenase (LDH) leakage, neutral red uptake, formazan formation, and crystal violet staining assays. HTP-WCSCs exhibited lower overall cytotoxic potency than 3R4F-WCSC when normalized to total particulate matter or nicotine concentration. Notably, the LDH leakage assay consistently yielded lower toxicity estimates, likely due to non-lytic cell detachment. Despite the difference in cytotoxic potential, HTP-WCSCs and 3R4F-WCSC triggered similar cytotoxic responses; focal adhesion disruption, cell detachment, and subsequent anoikis, potentially mediated by Bit1. This detachment-induced cell death was associated with thiol depletion and reactive oxygen species generation. In vivo relevance of these findings was confirmed in the lungs of rats exposed to 30 mg/mL Lil-WCSC or 200 µg/L cigarette smoke via respiratory routes for 2 or 4 weeks, respectively. Despite quantitative differences in cytotoxicity, the mechanisms and patterns were consistent across all products. These results suggest that smoking, irrespective of product type, may compromise respiratory epithelial integrity through the induction of anoikis.

Antineoplastic agents are hazardous compounds frequently used in cancer treatment. It is already known that the hospital environment poses a risk of occupational exposure to these agents. However, recent years, the rise of outpatient treatment and at-home treatment has introduced an additional risk including also cohabitants of patients. We identified a clear need for highly sensitive monitoring methods to assess exposure to high-risk compounds in a home setting. This study presents two validated methods for quantifying five high-risk antineoplastic agents in urine: one for cyclophosphamide, etoposide, mitomycin C and imatinib, and one for alpha-fluoro-beta-alanine. Liquid-liquid extraction with ethyl acetate was used for extraction of cyclophosphamide, etoposide, mitomycin C and imatinib from urine. Alpha-fluoro-beta-alanine was extracted using solid-phase extraction with Oasis HLB cartridges. All samples were analysed using ultra-performance liquid chromatography coupled to tandem mass spectrometry. During method validation, selectivity, extraction efficiency, matrix effect, process efficiency, linearity, sensitivity, precision and accuracy were established. The lower limits of quantification were determined to be 0.1 ng/mL (cyclophosphamide and mitomycin C), 0.7 ng/mL (etoposide), 1 ng/mL (alpha-fluoro-beta-alanine) and 10 ng/mL (imatinib). The methods were fully validated and are now ready for application in the field.