Introduction Chlordiazepoxide is used to manage alcohol withdrawal symptoms. Owing to its active metabolites with long half-lives, vulnerable patients may be at increased risk of prolonged sedation and respiratory depression, potentially requiring intensive care. This study aimed to characterize chlordiazepoxide dosing and related plasma concentrations in patients admitted to an intensive care unit or high dependency unit after chlordiazepoxide treatment for alcohol withdrawal symptoms. Methods A prospective clinical cohort study of patients admitted to the intensive care unit or high dependency unit after protocolized treatment with chlordiazepoxide ≥200 mg. Patients were classified as either sedated (respiratory insufficiency or somnolence) or agitated (alcohol withdrawal symptoms despite benzodiazepine treatment). Blood samples were collected at enrolment and 12 h later to measure concentrations of chlordiazepoxide, norchlordiazepoxide, and demoxepam. Results Of 26 patients included, 19 patients were sedated, and seven were agitated. At inclusion, 85% had plasma chlordiazepoxide concentrations above 3.5 mg/L. The median cumulative dose was higher in the agitated group (1,350 mg [interquartile range: 900–2,050 mg]) compared to the sedated group (700 mg [interquartile range: 275–1,250 mg]). Plasma chlordiazepoxide concentrations were significantly higher (P = 0.002) in the agitated group compared to the sedated group (20 mg/L versus 8.6 mg/L). Correlation between dose and plasma concentration (r 2 = 0.35; P = 0.001) was poor. Discussion Agitated patients were treated with higher doses and expectedly had higher chlordiazepoxide plasma concentrations than sedated patients, indicating that individual sensitivity and other differences play a role in clinical outcome, which emphasizes that therapeutic drug monitoring is not suitable for guiding treatment with chlordiazepoxide. Conclusions Poor correlation between doses, plasma concentrations, and clinical effects limits the ability to use cumulated doses and plasma concentrations to predict the risk of prolonged sedation and respiratory depression, making chlordiazepoxide unreliable in the treatment of alcohol withdrawal symptoms.

This study presents the first complete chloroplast genome assembly of Calophyllum membranaceum Gander & Champ 1849, a species noted for its pharmacologically active metabolites. The chloroplast genome of C. membranaceum measures 160,820 bp in length and an overall GC content of about 36.4%. It contains a total of 131 genes, which consisted of 86 protein-coding genes, 37 tRNAs, and 8 rRNAs. The phylogenetic analysis based on chloroplast data from the Malpighiales order suggests a close relationship between C. membranaceum and C. brasiliense. Our results provide valuable genomic resource that support future research on genetic resources, conservation, and medicine.

Auditory steady-state responses (ASSRs), particularly at 40 Hz, are promising biomarkers for psychiatric disorders involving dysregulated neural synchronization. While most ASSR studies have focused on the glutamatergic system, the serotonergic system, specifically 5-HT2A receptor signaling, has received limited attention. Psilocin, the active metabolite of psilocybin and a known 5-HT2A receptor agonist, alters cortical oscillatory activity, but its effects on ASSR dynamics remain unclear. In this study, we examined psilocin's effects on ASSRs in eight adult male Wistar rats implanted with 21 cortical electrodes. The rats were exposed to 40 Hz and 80 Hz click-train stimulation before and 30 minutes after subcutaneous psilocin administration (4 mg/kg). EEG signals were analyzed using time-frequency decomposition to extract phase-locking index (PLI) and event-related spectral perturbation (ERSP) values from frontal and temporal regions of both hemispheres. Psilocin selectively decreased PLI at 40 Hz stimulation in the right temporal cortex, with no significant changes in the frontal or left temporal regions, nor in response to 80 Hz stimulation. ERSP analysis revealed a global reduction in spectral power after psilocin administration in response to 80 Hz stimuli, but no consistent effects at 40 Hz. These results indicate that psilocin induces region- and frequency-specific alterations in auditory neural synchronization, characterized by right-lateralized disruption of 40 Hz phase-locking. This highlights the sensitivity of low-gamma oscillations to serotonergic modulation and supports the use of ASSR paradigms in translational models of altered perceptual and cognitive states.

Integrated multi-omics approaches and network pharmacology analysis to explore the diagnosis and regulatory role of active metabolites in allergy.

Atrazine exposure during puberty causes long-lasting neurochemical alterations and sex-dependent sexual behavior deficits in rats.

Increased TGF-β signaling during antiestrogen therapy in triple-negative breast cancer cells.

Mechanistic PBPK/PD modeling of free and nano-liposomal irinotecan reveals formulation-specific determinants of disposition and efficacy.

Corydalis Decumbentis Rhizoma (CDR) is a traditional Chinese herbal medicine, which has been clinically used for treating migraine, relieving the symptoms of rheumatoid arthritis, and reversing osteoporosis. Bicuculline (BIC) is one of the main bioactive ingredients present in CDR. The purpose of this study was to comprehensively investigate the interaction of BIC with cytochrome P450 enzymes and the alteration of the pharmacokinetic properties of the active thiol metabolite 4 of clopidogrel when coadministered. BIC showed time-, concentration-, and NADPH-dependent inhibitory effects on CYP2C19. BIC (2.0 μM) inhibited CYP2C19 activity by approximately 64% after 9 minutes of incubation at 30 °C, and KI and kinact were 0.09 μM and 0.051 min-1, respectively. Ticlopidine showed a significant protective effect against BIC-induced CYP2C19 inactivation. Dialysis did not restore the inhibited enzyme activity induced by BIC. Moreover, the addition of catalase/superoxide dismutase or glutathione did not show protective effects against BIC-induced enzyme inactivation. It is speculated that carbene intermediates were involved in BIC-induced inactivation of CYP2C19 because K3Fe (CN)6 restored the enzymatic activity. In addition, BIC and CDR extract pretreatment resulted in a significant decrease in the Cmax and area under the curve of the plasma thiol metabolite in rats given clopidogrel. SIGNIFICANCE STATEMENT: This study identifies bicuculline, a major component of Corydalis Decumbentis Rhizoma, as a mechanism-based inhibitor of CYP2C19 through carbene-mediated coordination with heme iron. Such inhibition reduces clopidogrel activation and indicates clinically relevant herb-drug interactions.

Tracking production and interconversion of extra- and intra-cellular metabolites during beer fermentation by ramanomics.

The Akkermansia muciniphila-tryptophan metabolism-aromatic hydrocarbon receptor axis mediates the protective effect of Schisandra chinensis pectin polysaccharide against colitis.

SuHeXiang Wan in the treatment of stroke: Prediction potentially active metabolites using a combination of in silico analysis and experimental viability assessment.

Fenofibrate anti-RARα/RXRα dimerization to attenuate all-trans retinoic acid-induced hyperlipidemia and hepatic steatosis in mice.

Daily oral cyclobenzaprine hydrochloride (HCl) has provided transient benefits in fibromyalgia, a chronic pain condition. To improve this effect, we evaluated sublingual formulations designed to drive transmucosal absorption. Two open-label studies evaluated the pharmacokinetics (PK), tolerability, and relative bioavailability of sublingual cyclobenzaprine HCl in healthy adults. In Study 1 (n = 24), three 2.8mg sublingual formulations of cyclobenzaprine HCl containing potassium phosphate dibasic (A), sodium phosphate dibasic (B), or trisodium citrate (C) were compared to immediate release (IR) cyclobenzaprine HCl 5mg. All sublingual formulations showed increased bioavailability (154% [A], 126% [B], and 125% [C]) and rapid absorption. Formulation A demonstrated the most favorable PK, with a ∼3min absorption lag versus ∼37min for oral IR, and a 783% higher dose-normalized AUC0-1. Formulation A was designated TNX-102 SL for further development. In Study 2 (n = 16), TNX-102 SL 2.8 and 5.6mg exhibited dose proportionality and no food effect. Furthermore, this is the first report describing the active metabolite norcyclobenzaprine in clinical studies, showing an elimination half-life of ∼60 h. Oral hypoesthesia and abnormal taste were the most common adverse events. These findings support TNX-102 SL as a rapidly absorbed and efficient sublingual tablet formulation of cyclobenzaprine HCl, providing effective transmucosal delivery.

Simultaneous quantification of acetylsalicylic acid, clopidogrel, ticagrelor and their major metabolites in human plasma by liquid chromatography-tandem mass spectrometry.

Plasmid-transformed Bifidobacterium longum 105A secreting β-glucuronidase for prodrug conversion of SN-38 glucuronide.

Gastric cancer, the fifth most prevalent cancer globally, poses significant treatment challenges due to factors such as late diagnosis, early metastasis, limited surgical options, and the systemic toxicity of chemotherapy. Because luminal barriers are often compromised in gastric cancers , orally administered therapies that enable localized absorption and drug release represent a promising new direction for site-specific treatment with limited side effects. We introduced disulfide-linked thermostable exoshell system that orally delivered protein-based bioorthogonal catalytic centres directly to cancer tissues. The highly engineered exoshells effectively encapsulated and stabilized labile catalytic centres, preventing degradation in the harsh gastric environment. In vivo gastric tumors were treated using the anti-cancer properties of active metabolites of the prodrug indole-3-acetic acid (IAA) converted in situ via bioorthogonal catalysis. In vitro cell studies revealed a dose- and time-dependent inhibition of gastric cancer cell growth, irrespective of their HER2 status. This inhibition was accompanied by upregulation of mitochondrial lipid peroxidation, reduced mitochondrial membrane potential, and activation of necroptotic pathway markers such as RIP1, RIP3, and MLKL at both mRNA and protein levels. In a mouse model of gastric cancer induced by N-Methyl-N-Nitrosourea, oral administration of catalytic exoshells for 6 weeks significantly inhibited gastric inflammation and tumour polyp growth. Additionally, LC/MS/MS-based metabolomic analysis of plasma obtained from treated mice showed significant upregulation of cytotoxic metabolites of IAA. Notably, metabolites relevant to redox regulation, including alpha-tocopherol (vitamin E), glutathione (GSH), homocysteine, methyl cysteine, and cysteine sulfinic acid, were identified as the top differentially expressed metabolites, indicating potent suppression of inflammation and tumour growth. Histological analysis of gastric tissue showed a reduced number of polyps and subsequent development of gastric tumours. Our in vitro and in vivo results demonstrated that exoshells possessed significant potential as an orally administered, titratable therapeutic platform for the management of gastrointestinal cancers.

Global warming poses a public health risk, and heat-induced reproductive defects are a growing concern for both humans and animals. Disrupted estrous cycle and reduced estrogen synthesis are notable changes in females exposed to heat stress; however, the underlying mechanism remains largely unknown, thus effective preventive or interventional strategies are still lacking. Here, by focusing on Cyp19a1, the gene encoding the rate-limiting enzyme for estrogen synthesis in ovarian granulosa cells, we identify heat stress as a trigger for Ca2+ release from the endoplasmic reticulum (ER). The increased intracellular Ca2+ flux functions as a high-temperature sensor and activates STAT3 phosphorylation. The activated Ca2+-pSTAT3 cascade disrupts the expression of H3K27me3-modifying enzymes, thus elevating H3K27me3 levels and finally represses Cyp19a1. Moreover, we demonstrate that retinoic acid (RA), the primary active metabolite of vitamin A, rescues CYP19A1 expression by antagonizing heat stress on multiple layers of the cascade. In vivo, RA administration rescues estrogen synthesis and corrects the estrous cycle in female mice under heat stress. Together, our study establishes a mechanistic link between heat stress and impaired estrogen synthesis and identifies the antagonistic function of RA. Finally, we propose a nutritional strategy to prevent or alleviate high temperature-induced estrus disorders in females.

The nucleoside analogue cytarabine (ara-C) is part of standard treatment against acute myeloid leukaemia (AML). The efficacy of this therapy is dependent upon accumulation of the active triphosphate metabolite ara-CTP, which mis-incorporates into genomic DNA, triggering cell death. The deoxyribonucleoside triphosphate triphosphohydrolase (dNTPase) SAMHD1 can hydrolyse ara-CTP and thereby convert the active metabolite back to its inactive prodrug form. This constitutes a barrier to treatment efficacy and thus strategies to target SAMHD1 are warranted. SAMHD1 activity is allosterically regulated by nucleotides, which are synthesised in cells via distinct pathways. We screened a collection of drugs targeting nucleotide biosynthetic enzymes and identified that inhibition of inosine-5'-monophosphate dehydrogenase (IMPDH), responsible for catalysing the rate-limiting step in guanine nucleotide biosynthesis, sensitises AML cell lines to ara-C in a SAMHD1-dependent manner. We show that approved drugs inhibiting IMPDH-mycophenolic acid and ribavirin-imbalance deoxyribonucleoside triphosphate pools and increase ara-C efficacy in SAMHD1-proficient, but not deficient, leukaemic cells. Altogether, we provide insight into SAMHD1 regulation in leukaemic cells and show how this process can be exploited by approved drugs to improve ara-C therapy.

Metabolic dysfunction-associated steatotic liver disease (MASLD) has become a major global public health problem, and its occurrence is associated with adverse environmental exposures during development. In this study, we investigated the impact of the use of prednisone (a synthetic glucocorticoid drug) during pregnancy on susceptibility to MASLD in offspring and explored its potential therapeutic targets. Pregnant rodents were administered clinically equivalent doses of prednisone daily by oral gavage during gestation days (GDs) 0-20 in rats and GD0-18 mice, and their offspring were fed a high-fat diet from postnatal weeks 8-12. The results showed that prenatal prednisone exposure (PPE) led to reduced hepatic glucose uptake and fatty acid oxidation in offspring rats prenatally and postnatally and that the offspring developed more severe MASLD when fed a high-fat diet, with males exhibiting greater severity than females. Consistent findings were observed in PPE adult offspring mice. RNA-seq and experimental results revealed that hepatic Serpina3c expression was consistently reduced in PPE offspring before and after birth, which led to an increase in chymase-Ang II production and subsequent activation of its receptor AT1R, leading to MASLD susceptibility. In vivo and in vitro studies revealed that the programming of low Serpina3c expression was associated with reduced H3K27ac levels in the gene promoter region of Serpina3c caused by the activation of GR-HDAC3 signaling by the active metabolite prednisolone. Finally, postnatal high expression of hepatic Serpina3c reversed the activation of the chymase-Ang II-AT1R pathway and significantly ameliorated hepatic glucose and lipid metabolic dysfunction and MASLD susceptibility in PPE offspring. In summary, this study reveals MASLD susceptibility in offspring induced by PPE and identifies Serpina3c as a target for the prevention and treatment of MASLD susceptibility.

Abstract Background: Mutations in the estrogen receptor gene (ESR1), particularly Y537S and D538G, are key drivers of resistance in estrogen receptor-positive (ER+) metastatic breast cancer (MBC) following aromatase inhibitor therapy. These mutations promote ligand-independent ER activation. In this in silico modeling study, we examined the biophysical behavior and downstream transcriptional impacts of (Z)-endoxifen, an active tamoxifen metabolite, a selective estrogen receptor modulator (SERM), across ESR1 variants to further elucidate its mechanism of action and therapeutic potential. Methods: We conducted full-atom molecular dynamics (MD) and metadynamics simulations of ERα-ligand complexes (wild-type and mutant) to assess conformational dynamics and antagonist state probability. Alchemical Non-Equilibrium Switching (NES) simulations compared binding affinities of (Z)-endoxifen vs. 4-hydroxy-tamoxifen (4-OHT). Luciferase reporter assays and 2D growth assays were performed in 293T and MCF-7 cells expressing ESR1-WT or D538G. Transcriptomic data were analyzed from public datasets, including RNA-seq and microarray profiles from ESR1 mutant tumors (n=56) and drug-treated breast cancer cells (n=27 for (Z)-endoxifen; n=5 for elacestrant). Differential expression and enrichment analyses (gene sets, pathways, TFs) were conducted using the limma and gseapy packages. Results- Biophysical Modeling: MD simulations (4 μs) showed stable (Z)-endoxifen binding across WT and mutant ERα, with no spontaneous receptor reactivation. Alchemical free energy calculations revealed favorable or equivalent binding of (Z)-endoxifen vs. 4-OHT (ΔΔG = 0.09 kcal/mol for WT; 1.30 kcal/mol for D538G). Metadynamic simulations on apo ERα indicated that mutations skewed toward the active state but did not prevent (Z)-endoxifen from stabilizing the antagonist conformation (antagonist state probability: WT 93.5%, Y537S 35.4%, D538G 46.9%). Functional Assays: (Z)-endoxifen potently suppressed ER-Luc activity (>70% inhibition) in 293T cells expressing ESR1-WT and all tested mutants (Y537S, Y537N, D538G, K303R), showing comparable potency across constructs. In MCF-7 cells, (Z)-endoxifen significantly reduced proliferation in both parental and ESR1-D538G lines. D538G expression did not rescue growth; a 60% reduction in 2D proliferation was observed with treatment. Transcriptomic Analysis: Gene expression analyses identified numerous transcripts with reversed regulation between ESR1 mutant tumors and (Z)-endoxifen-treated cells, indicating a functional reversal of mutant-driven expression patterns. Pathways upregulated in ESR1 mutant tumors and downregulated by (Z)-endoxifen included estrogen response, E2F targets, and Myc targets. Conversely, oxidative phosphorylation and p53 signaling were suppressed in ESR1 mutants and upregulated by (Z)-endoxifen (FDR < 0.05).(Z)-endoxifen also reactivated suppressed TF target networks (e.g., AHRR, GLIS2, NFATC4) in mutant tumors. In contrast, elacestrant had minimal transcriptional impact in ESR1 mutant MCF-7 cells, suggesting a narrower effect on disease-relevant pathways. Conclusion: (Z)-Endoxifen demonstrates robust biophysical and functional activity against ESR1 mutations. It stabilizes inactive ERα conformations and reverses mutant-driven transcriptional programs, showing greater transcriptional breadth than next-generation SERDs like elacestrant. These findings support (Z)-endoxifen’s potential as a precision therapy for ESR1-mutant ER+ MBC and justify further clinical investigation in endocrine-resistant settings. Citation Format: S. S. Hammer, E. Kirilin, H. de Almeida, V. Aladinskiy, A. Ustiugova, A. Shneyderman, A. Veviorskiy, M. Korzinkin, S. C. Quay. (z)-endoxifen maintains erα antagonist function against esr1 mutants via inactive conformation stabilization and reversal of mutant esr1-associated transcriptional signatures [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2025; 2025 Dec 9-12; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2026;32(4 Suppl):Abstract nr PD10-05.