Frontiers in Pharmacology最新文献

Pro-inflammatory and pro-thrombotic stimuli can activate endothelial cells (ECs) and predispose them to thrombotic microangiopathies (TMAs). Drug-induced TMA (DITMA) may occur in clinical practice during treatment with interferon-β1a (IFN-β1a), ciclosporin A (CsA), and gemcitabine (GEM). DITMA may also trigger the complement system and induce membrane attack complex (MAC, C5b-9) deposition in vivo, although their role and the benefit of inhibition remain unclear. In an experimental in vitro model of microvascular ECs exposed to these three drugs, we searched for MAC deposits and drug-specific pro-inflammatory and pro-thrombotic traits to gain insights into the mechanisms potentially involved in DITMA. Human microvascular endothelial cells line-1 (HMEC-1) was treated with 10% normal human serum, CsA, GEM, and IFN-β1a. Cell viability for each drug was measured using the resazurin assay. Cell component expression of the following markers involved in endothelial pathogenic activation was measured via immunofluorescence and flow cytometry: C5b-9, interleukin (IL)-1α, IL-6, E-selectin, platelet EC adhesion molecule-1 (PECAM-1), intercellular adhesion molecule-1 (ICAM-1), and von Willebrand factor (vWF). Levels of plasminogen activator inhibitor-1 (PAI-1) and urokinase plasminogen activator (uPA) were measured in the supernatants using the enzyme-linked immunosorbent assay (ELISA). Significantly increased C5b-9 deposits were found with each drug, and increased drug-specific activation marker expressions appeared in HMEC-1s when exposed to CsA (IL-1α, IL-6, ICAM-1, E-selectin, vWF, and uPA), GEM (IL-1α, IL-6, PECAM-1, ICAM-1, E-selectin, and vWF), and IFN-β1a (PECAM-1, ICAM-1, PAI-1, and uPA). Each drug induces MAC deposits on HMEC-1s and singular endothelial activation profiles, potentially leading to thrombogenesis observed in DITMA.

Colorectal cancer (CRC) patients with a microsatellite-stable (MSS) status exhibit poor responsiveness to PD-1/PD-L1 blockade. Pyroptosis induction may resensitize MSS tumors to PD-1/PD-L1 blockade; however, the expression of GSDME, a key executor of pyroptosis, is often downregulated in CRC. Here, curcumin (CUR), a natural polyphenol, was identified as a potentiator of GSDME-dependent pyroptosis in CRC. We discovered that CUR upregulates GSDME expression by inhibiting the ubiquitin-proteasome system (UPS) in the MSS-type CT26 and HT29 cell lines and activating the caspase-3/GSDME signalling axis, resulting in increased pyroptosis. In CT26 tumors, CUR-enhanced pyroptosis reshaped tumor-infiltrating immune subsets and potentiated the efficacy of anti-PD-1 therapy. Notably, the synergistic antitumor activity of CUR combined with PD-1 blockade in CT26 tumors is strictly dependent on the caspase-3/GSDME axis, as the therapeutic benefit was abolished in GSDME-knockout tumors. These findings establish CUR as a safe and effective adjuvant for PD-1/PD-L1 blockade in MSS CRC, particularly in tumors with low GSDME expression.

Objective: To develop and validate a population pharmacokinetic/pharmacodynamic (PK/PD) model for ciprofol in elderly surgical patients, delineating its pharmacokinetic profile and concentration-effect relationship to inform precision dosing.

Methods: Twenty patients (aged ≥65 years) undergoing elective surgery were enrolled. We performed population PK/PD analysis using nonlinear mixed-effects modeling on 386 arterial blood samples and synchronized Bispectral Index (BIS) data. A linear three-compartment model and a sigmoid Emax model described the PK and PD (BIS), respectively. Covariates (age, weight, gender, and laboratory parameters) were tested via stepwise selection. Model performance was evaluated using goodness-of-fit plots, bootstrap (n = 1,000), and prediction-corrected visual predictive checks. Dosing regimens were optimized via Monte Carlo simulation.

Results: A three-compartment model best described the PK. The center volume (V₁) was generally approximated at 2.95 L, but the peripheral volumes (V₂ and V₃) were 45.15 L and 76.79 L, respectively. The clearance (CL) was assessed at 1.01 L min^-1. Body weight and age significantly influenced CL. PD analysis showed rapid effect-site equilibration (K_e0: 1.09 min^-1), with an EC₅₀ of 233.91 ng mL^-1 and a Hill coefficient of 3.00. No covariates significantly affected PD parameters. The model exhibited sufficient fit and strong predictive efficacy. The simulation results confirmed that administering an intravenous loading dose of 0.4 mg kg^-1 over 1 min, followed by an initial continuous infusion at a rate of 0.6 mg kg^-1·h^-1 for 2 h, could stably maintain the patients' BIS values within the target range of 40-60.

Conclusion: A population PK/PD model for ciprofol in elderly patients was successfully established and validated. The model supports optimized, individualized dosing to achieve target anesthesia depth in this population.

The core pathophysiological mechanism of type 2 diabetes mellitus (T2DM) is closely associated with gut microbiota dysbiosis and its consequential impairment of enteroendocrine glucagon-like peptide-1 (GLP-1) secretion. T2DM mouse model was established using high-fat diet (HFD) feeding combined with streptozotocin (STZ) administration. Diabetic mice received 30 or 60 mg/kg of leonurine (LEO) via daily gavage for 12 weeks. Gut microbiota composition was profiled by metagenomic sequencing, fecal short chain fatty acids (SCFAs) concentrations were quantified via enzyme-linked immunosorbent assay (ELISA), and GLP-1 expression was assessed using oral glucose tolerance tests (OGTT), ELISA, and immunofluorescence. In vitro, high-glucose (25 mM)-challenged GLUTag enteroendocrine cells were employed to delineate the butyrate-mitochondrial pyruvate carrier 2 (MPC2) regulatory network using qPCR and Western blotting. LEO intervention significantly ameliorated glucose intolerance in diabetic mice and elevated GLP-1 levels in serum and colonic tissues. Metagenomic analysis revealed that LEO (60 mg/kg) remodeled gut microbiota structure, markedly enhancing α-diversity and specifically enriching butyrate-producing Alistipes. Mechanistically, butyrate activated MPC2 expression, effectively restoring cristae architecture defects observed by transmission electron microscopy, thereby promoting GLP-1 secretion. Crucially, MPC2 knockdown abrogated the secretagogue effect of butyrate on GLP-1 in GLUTag cells. LEO alleviates T2DM by remodeling the gut microbiota ecosystem, enhancing butyrate biosynthesis, and activating an MPC2-dependent mitochondrial energy metabolism pathway to reverse GLP-1 secretory dysfunction in intestinal L cells. This study establishes MPC2-mediated mitochondrial functional repair as a core mechanism through which microbial metabolites regulate enteroendocrine hormone secretion, identifying a novel therapeutic target within the "gut-islet axis" for diabetes intervention. Future studies should identify its active constituents, elucidate downstream effectors, and validate this mechanism in germ-free models.

In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantitative determination of amantadine in human plasma, with the incorporation of an internal standard to improve analytical accuracy. Plasma samples collected from volunteers were processed using acetonitrile-methanol (3:1, v/v) as the extraction solvent, followed by protein precipitation and purification via the QuEChERS (Quick, Easy, Cheap, Efficacious, Rugged, and Safe) method. Analysis was performed using LC-MS/MS under multiple reaction monitoring mode, with a total run time of 8 min. Quantification was carried out using the internal standard method. After a single oral administration of 200 mg amantadine hydrochloride, plasma concentrations were measured at various time points. Pharmacokinetic parameters were derived by fitting the data to a pharmacokinetic model using specialized software. The results demonstrated good linearity over the range of 0.5-20 ng/mL, with a correlation coefficient (R²) of 0.9978. The extraction recovery ranged from 94.5% to 110.1%, and both intra-day and inter-day relative standard deviations (RSD) were below 10%. The limit of detection (LOD) and limit of quantification (LOQ) were 0.15 ng/mL and 0.5 ng/mL, respectively. The absorption and elimination processes of amantadine in plasma followed first-order kinetics, with R² > 0.9. Notably, gender-specific differences were observed in the time to maximum concentration (T_max) and maximum concentration (C_max): females achieved a C_max of 670.23 ng/mL at 4 h, whereas males reached a C_max of 650.87 ng/mL at 8 h. This LC-MS/MS method is simple, rapid, and accurate, rendering it suitable for pharmacokinetic studies of amantadine in humans. Additionally, the established kinetic model provides valuable references for clinical medication guidance.

Background: Attention-deficit/hyperactivity disorder (ADHD) is a prevalent neurodevelopmental disorder in children. Long Mu Ning Xin Decoction (LMNXD) shows established clinical efficacy against ADHD, yet its mechanistic basis is not fully elucidated.

Objective: This study investigates the therapeutic potential of LMNXD for ADHD and explores its underlying mechanisms of action.

Methods: Thirty spontaneously hypertensive rats (SHRs/NCrl) were randomly divided into five groups: a model (SHR) group, low-, medium-, and high-dose LMNXD (LMNXD-LD, LMNXD-MD, LMNXD-HD)groups, and a methylphenidate hydrochloride (MPH) group. Additionally, six Wistar Kyoto (WKY/NCrl) rats were designated as the control group.Behavioral performance was assessed using the open field test and Morris water maze. The expression levels of glial fibrillary acidic protein (GFAP), dopamine deceptor D1 (DRD1), and brain-derived neurotrophic factor (BDNF) in the rat hippocampus, prefrontal cortex (PFC), and striatum were evaluated by immunofluorescence, immunohistochemistry, and Western blot. Potential targets and mechanisms were explored through transcriptomic sequencing and network pharmacology, with subsequent validation by reverse transcription quantitative polymerase chain reaction (RT-qPCR).

Results: Compared to the SHR group, LMNXD ameliorated hyperactivity, impulsivity, deficits in spatial memory and learning ability in SHR/NCrl rats. It also effectively reduced GFAP expression in the hippocampus while increasing DRD1 expression in the PFC and BDNF levels in the striatum. Network pharmacology predicted that LMNXD might alleviate ADHD by acting on pathways including phosphatidylinositide 3-kinase-Akt (PI3K-Akt), calcium signaling, and cyclic adenosine monophosphate (cAMP) signaling. Consistent with this prediction, transcriptomic analysis of rat hippocampi showed that LMNXD influences the cAMP and PI3K-Akt signaling pathways, as well as serotonergic and cholinergic synapses. RT-qPCR further confirmed that LMNXD likely exerts its therapeutic effect by regulating the mRNA expression of ATPase Plasma Membrane Ca²⁺ Transporting 4 (ATP2B4), Glutamate Ionotropic Receptor NMDA Type Subunit 3A (GRIN3A), Oxytocin Receptor (OXTR), Collagen Type VI Alpha 2Chain (COL6A2), and Integrin Subunit Alpha 1 (ITGA1) within the cAMP andPI3K-Akt pathways.

Conclusion: LMNXD may ameliorates hyperactive-impulsive behaviors and improves spatial memory and learning in SHRs/NCrl rats by modulating ATP2B4, GRIN3A, OXTR, COL6A2, and ITGA1 within the cAMP and PI3K-Akt signaling pathways. This intervention also upregulates DRD1 and BDNF expression while downregulating GFAP levels.

Background: The intensive use of non-steroidal anti-inflammatory and analgesic drugs (NSAIDS) worldwide poses a challenge to scientists because of the adverse side effects. This article aims to synthesize a novel group of 1-aminoindazole-isatin Schiff base compounds considering their potency as analgesic and anti-inflammatory agents.

Methods: The synthesis of novel agents involved reflux condensation of isatin derivatives (5 mmol) and 1-aminoindazole (5 mmol) in ethanol for 2 h, which were then characterized for their structural integrity. In silico evaluation using PyRx, BIOVIA Discovery Studio, and GROMACS was performed to determine the affinity of the specific receptors and compare them with the results gained by use of standard diclofenac before preclinical evaluation using albino mice (analgesic activity) and rats (anti-inflammatory activity). The preclinical analgesic potency was analyzed via Eddy's hot plate and tail-pin methods, whereas, the anti-inflammatory potency was analyzed through carrageenan-induced paw edema against diclofenac as the standard agent.

Results: A high percentage yield of the reactions was determined (≈80%); the IR, NMR, and mass spectra showed the compounds to be stable with no shifts, justifying the accuracy of the procedure employed. The molecular docking of the ligands with two different crystal structures of proteins of interest, i.e., COX-1 and COX-2, yielded stable and the lowest binding energies, i.e., -9.6 kcal/mol for AB 12 and - 7.1 kcal/mol for diclofenac. Through molecular dynamic simulations employing GROMACS for a time period of 50 ns, AB 12 and diclofenac also yielded a thermodynamically stable and structurally folded protein and ligand complex, showing an average of 0-3 (AB 12) and 0-5 (diclofenac) hydrogen bonds with the least system fluctuations and atom deviations; furthermore, the potential energy of the complete system was stabilized at an average point of - 685,000 kj/mol for both molecules. The preclinical results showed a significant value for the ligand AB 12 (p ≤ 0.01) against the diseased control group.

Discussion: The ligand AB 12, AB 14 and AB 15 is exceptional as an analgesic and anti-inflammatory agent. AB 12 further showed stable hydrogen bonds with protein COX-2 for 50 ns in comparison with diclofenac. Based on this study, these molecules can be considered best for future studies regarding the toxicological profile.

Escitalopram is widely regarded as a well-tolerated selective serotonin reuptake inhibitor (SSRI) with a favorable safety profile. However, severe adverse events can occur even at therapeutic doses in susceptible individuals. Here, we report a rare case of simultaneous life-threatening Syndrome of Inappropriate Antidiuretic Hormone secretion (SIADH) and cardiac toxicity induced by standard-dose escitalopram. A 51-year-old female (weight 50 kg) presented with severe fatigue and anorexia. Initial laboratory results revealed profound hyponatremia (116.1 mmol/L). Following sodium supplementation, serum sodium paradoxically decreased to 114.7 mmol/L ("desalination phenomenon"), while urinary sodium excretion was markedly elevated (220 mmol/24 h) alongside significant hypouricemia (76 μmol/L), confirming the diagnosis of SIADH. Concurrently, the patient manifested significant cardiac toxicity, including sinus bradycardia (41-55 bpm) and marked QTc prolongation (570 ms). Pharmacogenetic analysis identified the CYP2C19 *1/*2 genotype (Intermediate Metabolizer). Despite the therapeutic dosage (10 mg/day) and a non-toxic serum concentration (5 ng/mL measured 72 h post-discontinuation), the patient exhibited severe toxicity, likely driven by "phenoconversion" due to low muscle mass and physiological vulnerability, exacerbated by a pharmacodynamic synergism with low-dose quetiapine. Discontinuation of medications and strict fluid management resulted in complete resolution of both hyponatremia and arrhythmia. The causality was assessed as "probable" for both drugs using the Naranjo Algorithm, and the drug-drug interaction was rated as "probable" using the Drug Interaction Probability Scale (DIPS). This case highlights that genotype-phenotype mismatch, combined with pharmacodynamic synergism (escitalopram-quetiapine interaction), can precipitate severe neuro-cardiac toxicity even at therapeutic levels. It underscores that severe neuro-cardiac toxicity can occur even at therapeutic levels due to individual vulnerability. Therefore, routine monitoring of electrolytes and electrocardiograms (ECG) remains indispensable for patient safety, as pharmacogenetic screening and therapeutic drug monitoring may not predict such idiosyncratic reactions in resource-constrained settings.

Introduction: Ischemia is an important cause of acute kidney injury (AKI). Ischemia-induced hypoxia rapidly induces activation of the Jun amino-terminal kinase (JNK) in tubular epithelial cells of the kidney, and blockade of this enzyme is protective in short-term animal models of renal ischemia. However, the clinical translation of this finding requires a water-soluble JNK inhibitor. This study investigated whether KRev-202, a soluble prodrug of the potent and selective JNK inhibitor CC930, can prevent ischemia-induced AKI and whether short-term inhibition of JNK can prevent AKI from transitioning to renal fibrosis.

Methods: In a rat model of bilateral renal ischemia/reperfusion injury (IRI), the animals received prophylactic treatment with KRev-202, the parent compound (CC-930), or a vehicle by oral gavage, starting 1 h prior to surgery.

Results and discussion: In study 1, the animals were killed on day 1 after IRI to assess the AKI peak. Vehicle-treated animals exhibited a 4.5-fold increase in plasma creatinine levels, substantial tubular necrosis, increased tubular damage markers, and inflammation on day 1. Both KRev-202 and CC-930 treatment inhibited JNK activation, caused a 50% reduction in plasma creatinine levels, and substantially reduced tubular necrosis, tubular damage, and inflammation. In studies 2 and 3, treatments were administered from -1 h until day 4, and then the animals were killed on days 7 and 21, respectively. Compared to the vehicle group, a 4-day treatment with KRev-202 or CC-930 improved the recovery of tubular structure on day 7 and substantially reduced the development of renal fibrosis on day 21. Furthermore, KRev-202 treatment administered only during the first 24 h of IRI provided the same benefits as the 4-day treatment regimen, demonstrating the importance of early blockade of this pathway. In conclusion, KRev-202 is a new water-soluble JNK inhibitor with therapeutic potential for preventing ischemia-induced AKI.

Background: As a natural flavonoid, quercetin has anti-inflammatory and anti-oxidative activities. Studies confirm its beneficial effect on asthma, but the underlying mechanism remains unclear. This study aimed to systematically evaluate quercetin's efficacy in treating asthma, explore its regulatory role in asthma-related autophagy and associated signaling pathways, and provide new insights for asthma treatment research.

Methods: In vivo, ovalbumin (OVA)-induced asthma model mice were first successfully established, then randomly assigned to five groups: control, asthma model, low/high-dose quercetin, and dexamethasone positive control. ELISA, histopathological staining, immunohistochemistry and Western blot were used to assess quercetin's therapeutic effect and molecular mechanism. To complement the in vivo findings from the OVA-induced asthmatic mouse model, in vitro experiments were conducted using the human bronchial epithelial cell line BEAS-2B. Specifically, the cell line was stimulated with TNF-α and IL-4 to establish an inflammatory model, further validating quercetin's regulation of autophagy and inflammation.

Results: In vivo, quercetin reduced inflammatory cell count and proinflammatory cytokine levels in asthmatic mice's bronchoalveolar lavage fluid (BALF), lowered serum IgE, and alleviated lung inflammatory infiltration and pathological damage. It also inhibited lung autophagy and activated the PI3K/Akt/mTOR signaling pathway. In vitro, consistent with in vivo findings, quercetin downregulated proinflammatory factors and autophagy-related proteins in TNF-α/IL-4-stimulated BEAS-2B cells. In addition, the PI3K/Akt/mTOR signaling pathway is also activated by quercetin.

Conclusion: Quercetin attenuates airway inflammation and lung damage in asthmatic mice. Its therapeutic effect is associated with the modulation of PI3K/Akt/mTOR signaling pathway activity and the regulation of excessive autophagy, which provides new potential approaches and mechanistic insights for asthma treatment.