Brexpiprazole is a third-generation antipsychotic used for the treatment of schizophrenia or as an adjunctive drug for the treatment of affective and neurological disorders. The combined treatments with brexpiprazole and other drugs that are also cytochrome P450 (CYP) substrates may lead to pharmacokinetic drug-drug interactions. Our present work aimed to investigate the effects of prolonged administration of brexpiprazole on the expression of hepatic transcription factors and CYP drug-metabolizing enzymes. Male Wistar rats received brexpiprazole (1 mg/kg ip.) for two weeks. Their livers were excised 24 h after the last dose, and the activities (HPLC), protein levels (Western blotting), and mRNAs (qRT-PCR) of CYP enzymes were measured. In parallel, the expression of hepatic transcription factors (Western blotting, qRT-PCR) and the concentration of serum hormones (ELISA) were assessed. Brexpiprazole produced a broad-spectrum effect on CYP expression and activity. It enhanced the expression and activity of CYP1A, CYP2A, CYP3A1/2, and the activity of CYP2B, but decreased the expression/activity of CYP2Ds and CYP2E1. The observed changes in CYP enzymes corresponded to alterations in transcription factors: the increased expression of PXR and AhR, and decreased expression of PPARγ, LXR, and FXR. The above modifications in CYP enzymes' expression were accompanied by enhanced corticosterone and reduced T4 serum levels. Brexpiprazole affects the expression of hepatic transcription factors and CYP, which may impact its own biotransformation and the metabolism of endogenous substances and concomitantly administered drugs, and lead to drug-drug interactions of pharmacological/clinical importance.
The thymus plays a critical role in sustaining T-cell immunity, although its function is highly vulnerable to acute injury and physiologically declines with age, resulting in compromised immune responses. Impaired thymic function represents a major clinical challenge, particularly in settings of immunosuppression associated with cancer therapy and aging. Yet, effective strategies to rejuvenate the thymus remain limited. To explore novel regenerative approaches, we focused on FOXN1, a master regulator of thymic epithelial cell (TEC) development and function. By developing a custom screening platform, we tested a library of FDA-approved compounds for their ability to induce FOXN1 in TECs. Proteasome inhibition emerged as a potent and previously unrecognized mechanism for upregulating FOXN1 in both murine and human primary TECs. Among the hits identified in the screening, the antiparasitic drug nitazoxanide (NTZ) stood out for its proteasome inhibitory activity and for inducing Foxn1 expression while preserving cell viability, unlike other proteasome inhibitors. Mechanistically, NTZ-induced proteasome inhibition triggered endoplasmic reticulum stress (ER) and the adaptive unfolded protein response (UPR), ultimately engaging autophagy in TECs. In this context, the induction of autophagy acted as a compensatory mechanism to support cell survival in response to proteasome inhibition. Notably, when administered in mice, NTZ significantly accelerated functional thymic recovery after radiation-induced damage, promoting restoration of thymic architecture and cellularity of both stromal and hematopoietic compartments without disrupting physiological T-cell selection or tolerance mechanisms. Consistent with our in vitro findings, NTZ treatment induced Foxn1 and its downstream targets in TECs in vivo and conferred protection to TECs following irradiation. These findings uncover proteasome inhibition and, more broadly, modulation of ER stress and UPR pathways as a previously unrecognized mechanism regulating Foxn1 expression and position NTZ as a promising pharmacological strategy to enhance immunity in patients experiencing T-cell deficiencies due to cancer-related immunosuppression, infections, and age-related thymic atrophy.
Albumin-binding prodrugs represent a pharmacological strategy to improve systemic stability and tumor exposure of cytotoxic agents. Legumain, an asparagine endopeptidase selectively activated in the acidic tumor microenvironment, provides a mechanism for tumor-specific prodrug activation. In this study, we developed WC10-003, a legumain-responsive albumin-binding prodrug of belotecan, and investigated its pharmacological properties and antitumor activity in ovarian cancer models. WC10-003 was synthesized by conjugating belotecan to a legumain-cleavable peptide linker. Albumin association, plasma stability, and enzymatic activation were evaluated to characterize its pharmacological behavior. Antitumor efficacy was assessed in ovarian cancer xenograft models and compared with irinotecan and belotecan. Transcriptomic analysis was performed to explore treatment-associated signaling alterations, and immunohistochemistry and flow cytometry were used to evaluate changes in tumor-associated macrophage polarization. Following intravenous administration, WC10-003 rapidly associated with endogenous albumin, forming a stable circulating complex that prolonged systemic exposure and enhanced tumor drug accumulation. In ovarian cancer xenograft models, WC10-003 produced significantly greater tumor growth inhibition than irinotecan or belotecan, consistent with improved pharmacokinetic and tumor exposure profiles. Transcriptomic profiling revealed suppression of PI3K-AKT-related signaling pathways associated with tumor growth. In addition, immunophenotypic analyses demonstrated a shift in tumor-associated macrophages toward a less immunosuppressive phenotype. Combination treatment with WC10-003 and anti-PD-1 antibody further enhanced antitumor efficacy, suggesting that pharmacologically optimized prodrug activation can sensitize tumors to immune checkpoint blockade. WC10-003 exhibits favorable pharmacological properties, including albumin-mediated stabilization and legumain-dependent tumor activation, resulting in enhanced antitumor efficacy in preclinical ovarian cancer models. These findings highlight a pharmacologically driven prodrug strategy for improving cytotoxic drug performance and support its further evaluation in combination with immunotherapy.
BACKGROUND: Connexin-43 (Cx43) is the principal gap junction protein in the heart, mediating electrical coupling and ion exchange between cardiomyocytes to maintain synchronous contraction. Any disruption or malfunction of Cx43 can lead to arrhythmias and other cardiac issues. Understanding the functions and regulation of Cx43 is vital in both basic research and clinical contexts. Propafenone, a class Ic antiarrhythmic drug, has shown promise in rhythm control; however, its precise impact on cardiac cellular physiology, particularly regarding Cx43, remains incompletely understood. The present study investigated propafenone’s effects on Cx43 protein content, physiology, and underlying mechanisms in cell systems. METHODS: Cell lines include human embryonic kidney HEK293 cells transfected with Cx43 (Ex-HEK); differentiated murine embryonic carcinoma EPI7 cells with an epithelioid morphology and visceral endoderm-like END2 cells, both endogenously expressing functional Cx43. Cx43 protein contents were determined by Western blot analysis, whereas immunofluorescence (IF) imaging was used to assess the subcellular localization of Cx43 proteins. Dye injections were used to gain insight into the effects of propafenone on Cx43 function. RESULTS: Full-length Cx43 protein levels were dose-dependently increased after propafenone treatment and IF microscopy showed an intracellular accumulation of Cx43 protein, both on heterologously and endogenously expressed Cx43. Propafenone did not alter the Cx43 half-life, in contrast to the lysosomal inhibitor chloroquine. Finally, gap-junctional coupling was decreased by chronic propafenone treatment. CONCLUSION: We conclude that propafenone increases non-functional Cx43 protein content, resulting in its intracellular accumulation, as a side effect.
Hypoxic/ischemic brain injuries, including ischemic stroke and perinatal asphyxia, remain major causes of mortality and long-term neurological disability, establishing a demand for therapeutic strategies suitable against the multifactorial nature of underlying mechanisms. Estrogen receptors (ERs) signaling is known to exert neuroprotective effects, however, genomic ER activation is associated with serious adverse effects, including carcinogenesis and thromboembolisms. Pathway Preferential Estrogen-1 (PaPE-1), a compound that selectively activates the non-nuclear subset of ERs, may provide neuroprotection, thereby overcoming the deleterious effects. The aim of this study was to elucidate the molecular mechanisms underlying the neuroprotective effects of PaPE-1 in an in vitro model of hypoxic-ischemic neuronal injury, with particular emphasis on non-nuclear estrogen receptor signaling and its downstream pathways. Primary mouse cortical neuronal cells were subjected to 6 hours of experimental hypoxic/ischemic injury, followed by 18 hours of post-treatment with PaPE-1. Subsequently, a variety of biochemical assessments were conducted, including measurements of neuronal viability, cell death, and formation of autophagy-related vesicles. Moreover, the influence of PaPE-1 was assessed with molecular methods, encompassing measurements of gene and protein expression level and a set of epigenetic-related parameters, for instance, assessment of global DNA/RNA methylation and locus-specific methylation of genes and miRNA expression. To dissect signaling pathways, selective pharmacological inhibitors targeting mTOR/MEK1/2 and autophagy regulators were applied. ER subtype involvement was examined using ER-selective antagonists and specific siRNA silencing. Non-nuclear ER activation with PaPE-1 attenuated maladaptive autophagy, RNA/DNA oxidative stress damage, and neuronal degeneration while contributing to the regulation of gene expression and epigenetic processes. Evocation of robust neuroprotection involved modulation of mTOR and MEK1/2 signaling, predominantly mediated by estrogen receptor 1 (ESR1). In conclusion, PaPE-1 exhibits a multitarget mode of action that provides broad-spectrum protection against hypoxic/ischemic neuronal injuries. Considering its complexity of action and confirmed strong, neuroprotective activity, this compound holds promise for broader evaluation across different brain cell types and in vivo models.
The glutamatergic system, particularly N-methyl-D-aspartate (NMDA) receptors, has long been a significant focus of research into new strategies for treating depression, and the clinical success of ketamine, an NMDA receptor antagonist, has been a significant breakthrough. In parallel, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, another key component of the glutamatergic system, have been studied for decades, and renewed interest stems from evidence linking their activation to the rapid antidepressant effects and synaptic plasticity observed after ketamine administration. Among pharmacological agents targeting AMPA receptors, the class of positive allosteric modulators, AMPAkines, has attracted particular interest. These compounds act by prolonging AMPA receptor channel open time, thereby enhancing excitatory neurotransmission and upregulating brain-derived neurotrophic factor (BDNF) expression. Preclinical studies consistently demonstrated antidepressant-like effects of low-impact AMPAkines, which offer a favorable safety profile in contrast to high-impact compounds that carry seizure risk. Although preliminary clinical trials support these findings, their limited scope highlights persistent translational challenges. These include a narrow therapeutic window, suboptimal pharmacokinetic properties, and the limited predictive validity of animal models. AMPAkines thus represent a potentially promising class of rapid-acting antidepressants, although significant translational hurdles remain. This narrative review aims to synthesize evidence on the role of AMPA receptors in neuroplasticity, the therapeutic potential of AMPA receptor modulators (AMPAkines) in influencing neuroplasticity, and their potential therapeutic applications in depression. [Image: see text] Not applicable.
Combined therapy with epidermal growth factor (EGF) and growth hormone-releasing peptide 6 (GHRP6) has demonstrated neuroprotective effects in models of global and focal brain ischemia. Clinical studies in ischemic stroke patients have confirmed the safety and preliminary efficacy of this combined treatment. This study aimed to elucidate the molecular mechanisms underlying the effects of EGF+GHRP6 co-administration. Male Wistar rats were subjected to endothelin-1 (ET-1)-induced middle cerebral artery (MCA) occlusion and randomly assigned to three experimental groups: EGF+GHRP6‑treated ischemic, vehicle‑treated ischemic, and sham-operated controls (n = 12 per group). Label-free quantitative proteomic analysis of the ischemic penumbra was performed at 3 and 24 h post-treatment. Functional enrichment and pathway analysis of differentially modulated proteins were performed using bioinformatics tools. Proteomic profiling validated the ischemic model and revealed 40 and 223 proteins differentially modulated by EGF+GHRP6 at 3 and 24 h, respectively. Proteins involved in neurotransmitter transport were consistently overrepresented in the EGF+GHRP6-regulated proteome at both time points. At 24 h post-treatment, proteins associated with reactive oxygen species (ROS) detoxification, heat shock factor 1 (HSF1) activation, and negative regulation of cellular hypoxia response were significantly modulated. Additionally, anti-apoptotic and mitochondrial proteins were modulated in the ischemic penumbra, supporting the neuroprotective effects of EGF+GHRP6. Notably, proteins known to attenuate brain damage after stroke were up-regulated, while those promoting ischemic injury were down-regulated following the combined treatment. These findings provide molecular evidence supporting the neuroprotective mechanism of action of EGF+GHRP6 co-administration and reinforce its potential as a therapeutic strategy for ischemic stroke.
Depression in minors is a growing global health concern with limited pharmacological options. This study evaluated the safety profiles of four selective serotonin reuptake inhibitors (SSRIs)-fluoxetine, sertraline, escitalopram, and fluvoxamine- in children and adolescents, focusing on serious adverse events (SAEs). Pediatric adverse event reports from the FAERS database (Q1 2004-Q1 2025) were analyzed. Only reports listing the study drugs as Primary Suspect were included; duplicates were removed. Events were coded using MedDRA 26.1 and analyzed at system organ class (SOC) and preferred term (PT) levels. Disproportionality analyses used reporting odds ratio (ROR), proportional reporting ratio (PRR), Bayesian confidence propagation neural network (BCPNN), and multi-item gamma-Poisson shrinker (MGPS), with Bayesian shrinkage (EBGM05) applied to low-frequency signals. Psychiatric events, including suicidal ideation, suicide attempts, and self-injury, showed the most consistent signals. Fluoxetine and sertraline had broad profiles; escitalopram showed higher disproportionality for psychiatric, cardiac, and perinatal events; fluvoxamine showed elevated signals for impulsive behaviors and rare systemic events. Neurological events, such as serotonin syndrome and tremor, were consistently reported; congenital and perinatal signals had limited statistical stability. SSRIs share common psychiatric AE signals in minors, but differences in strength and distribution likely reflect real-world exposure, metabolic pathways, and structural properties rather than causal differences. FAERS-based pharmacovigilance provides a comparative overview of SSRI safety, with multi-algorithm assessment and Bayesian shrinkage enhancing low-frequency signal reliability, informing safer SSRI use and guiding future studies integrating pharmacokinetics and pharmacogenomics.
The thioredoxin system, comprising thioredoxin (Trx) and thioredoxin reductase (TrxR), is a central regulator of cellular redox homeostasis and plays essential roles in normal brain physiology and redox signaling. In glioblastoma (GBM), this system undergoes profound pathological rewiring, creating a redox dependency that represents a potential therapeutic vulnerability. The overexpression of Trx and TrxR in GBM promotes tumor proliferation, invasion, angiogenesis, and resistance to chemotherapy and radiotherapy, while the endogenous Trx inhibitor, thioredoxin-interacting protein (TXNIP), is frequently downregulated. This imbalance drives redox adaptation and sustains tumor survival under metabolic and therapeutic stress. Pharmacological modulation of the Trx system using synthetic inhibitors, such as auranofin, platinum-based compounds, and PX-12, as well as selected natural compounds including curcumin analogs and flavonoids, has shown efficacy in preclinical GBM models by inducing oxidative stress and enhancing sensitivity to standard therapies. Emerging evidence also suggests that Trx system targeting may modulate the tumor immune microenvironment, providing a rationale for combination strategies with immunomodulatory approaches. Overall, targeting the Trx system represents a promising precision oncology strategy for GBM. Future efforts should focus on the development of brain-penetrant inhibitors, rational combination therapies, and predictive biomarkers to facilitate clinical translation. Given the essential role of the Trx system in normal brain homeostasis, therapeutic targeting requires careful consideration of safety, therapeutic index, and tumor-selective vulnerabilities. This narrative review discusses current evidence on the physiological functions of the Trx system in the brain, its dysregulation in GBM, and its relevance as a precision therapeutic target.
Unilateral ureteral obstruction (UUO) induces oxidative stress, inflammation, ferroptosis, and progressive fibrotic remodeling. Whether pharmacological modulation of ferroptosis-related redox imbalance attenuates obstructive kidney injury remains unclear. In this study, we investigated the effects of the thiol-containing antioxidant 2-mercaptoethanol (2-ME) in a mouse UUO model. Mice subjected to UUO received either pre-treatment or delayed treatment with 2-ME. Ferroptosis-related markers, including glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), the ratio of reduced to oxidized glutathione (GSH/GSSG), and lipid hydroperoxides, as well as inflammatory mediators, F4/80-positive macrophage infiltration, Havcr1 mRNA expression, and fibrotic parameters were evaluated using molecular and histological analyses. UUO markedly decreased the expression of GPX4 and SLC7A11, reduced the GSH/GSSG ratio, and elevated lipid hydroperoxide levels. These changes were accompanied by increased tubular injury scores, infiltration of F4/80-positive macrophages, and extracellular matrix accumulation. Pre-treatment with 2-ME increased GPX4 and SLC7A11 expression, improved the GSH/GSSG balance, reduced lipid hydroperoxide levels, and attenuated inflammatory activation. Additionally, 2-ME pre-treatment significantly reduced tubular injury scores and Havcr1 mRNA expression. However, 2-ME did not consistently suppress collagen deposition or the expression of fibrosis-related genes. Delayed administration of 2-ME failed to significantly alter antioxidant, inflammatory, or fibrotic markers. Pre-treatment with 2-ME attenuates ferroptosis-associated redox imbalance and inflammatory responses in UUO but does not consistently suppress tubulointerstitial fibrosis. These findings suggest that 2-ME can serve as a pharmacological tool to modulate thiol-dependent redox balance and inflammatory activation during UUO, whereas fibrosis progression likely involves additional mechanisms. Not applicable.
Cerium oxide nanoparticles (CeO NPs) are showing neuroprotective effects in various experimental models of neurodegeneration. Doping of nanoparticles with magnetic resonance imaging (MRI) contrast agents (e.g., gadolinium) could enable simultaneous diagnosis and treatment of neurodegenerative diseases, a technology called theranostics that is used primarily in oncology but can also be successfully applied in the diagnosis and treatment of neurodegenerative diseases. In this study, we doped polyacrylic acid conjugated cerium oxide nanoparticles with gadolinium (Gd-CeO) to create a theranostic agent with MRI capabilities and neuroprotective properties. These nanoparticles were evaluated for their physicochemical characteristics, magnetic resonance imaging potential, biosafety profile, cellular uptake, and neuroprotective effects compared to CeO nanoparticles (CeO) in a human neuronal model of Parkinson's disease employing undifferentiated and retinoic acid-differentiated SH-SY5Y cells. The synthesized Gd-CeO nanoparticles showed good stability, concentration-dependent T1 and T2 contrast features, and were not cytotoxic. The Gd-CeO nanoparticles were rapidly taken by cells and maintained neuroprotective potency against hydrogen peroxide (H2O2)- and 6-hydroxydopamine (6-OHDA)-induced cell damage to a similar extent as did CeO nanoparticles without Gd doping. Moreover, we demonstrated a protective effect of Gd-CeO and CeO nanoparticles on mitochondrial membrane potential, DNA fragmentation, and the number of necrotic cells in both models of cell injury, whereas at the level of caspase-3 activity, we showed an inhibitory effect of the studied NPs only in the 6-OHDA model. Finally, the protection mediated by Gd-CeO and CeO nanoparticles against H2O2 was confirmed in mouse primary cortical neurons. Since the developed Gd-CeO nanoparticles showed promising contrast features, as well as maintaining biosafety and neuroprotective properties similar to those of nanoparticles without Gd doping, they could be further investigated as a potential theranostic probe for neurodegenerative diseases, including Parkinson's disease.
BACKGROUND: Developing semicarbazide-based compounds as potential therapeutic agents is a promising direction in medicinal chemistry, particularly in the context of anticancer drug design. This study reports the synthesis and comprehensive physicochemical characterisation of a series of novel semicarbazide derivatives (AW8, AW12, AW19, AW23, AW33, AW38). METHODS: Molecular structures and tautomeric forms in the crystalline state were elucidated using single-crystal X-ray diffraction and supported by detailed Fourier Transform Infrared Spectroscopy (FT-IR) analysis. Incorporation into 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)-based model lipid membranes was evaluated using attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and Langmuir monolayer techniques. Theoretical absorption, distribution, metabolism, and excretion (ADME) properties and in silico predictions were generated using SwissADME and ADMETlab 3.0 software platforms. Antioxidant potential was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging assays, and cytotoxicity against PANC-1 pancreatic cancer cells was determined using the 3-(4,5-di methyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: The compounds exhibited distinct hydrogen-bonding patterns, conformational preferences driven by substituent-dependent electronic effects, and characteristic supramolecular interactions governing their crystal packing, as confirmed by Hirshfeld surface analysis. Among the tested derivatives, AW8 and AW12 demonstrated the highest membrane affinity, inducing significant spectral alterations and surface pressure changes, indicative of enhanced membrane perturbation associated with chlorine substitution. Absorption, distribution, metabolism, excretion, and toxicity (ADMET) predictions revealed favourable pharmacokinetic profiles, including the potential for blood–brain barrier penetration in selected derivatives and predicted inhibition of P-glycoprotein, a key mediator of multidrug resistance. The compounds demonstrated moderate radical scavenging activity and dose-dependent cytotoxicity against PANC-1 cells. CONCLUSION: These findings indicate that the studied semicarbazides exhibit promising membrane-active properties, favourable ADME profiles, and biological activity relevant to anticancer drug development, supporting their further evaluation as potential therapeutic candidates.
Prostaglandin F2α receptor (FP receptor) signaling is a plausible target for promoting hair growth, but clinical data on topical latanoprost acid (the active free-acid FP agonist) in hair loss are lacking. This study aimed to evaluate the clinical efficacy, safety, and mechanistic basis of topical latanoprost acid in women with female androgenetic alopecia. In this investigator-initiated, randomized, double-blind, single-center, dose-ranging pilot trial, 29 adult women with hair loss predominantly consistent with female androgenetic alopecia were randomized to vehicle (n = 2) or topical latanoprost acid 0.01% (n = 8), 0.05% (n = 13), or 0.1% (n = 6), applied once daily for 6 months. The primary endpoint was within-participant change in target-area hair count (TAHC, hairs/cm²) from baseline to month 6; trichoscopic activity markers (yellow dots) and follicular-unit (FU) remodeling were secondary and exploratory outcomes. Human hair dermal papilla cells (HHDPCs) were assessed for FP receptor-linked signaling (intracellular Ca²⁺ flux) and DNA synthesis by 5-ethynyl-2'-deoxyuridine (EdU) incorporation after exposure to latanoprost acid versus equimolar latanoprost. An increase in TAHC was observed across all active treatment arms (mean ± SEM ΔTAHC: 17.8 ± 4.3, 23.5 ± 6.1, and 16.5 ± 6.5 hairs/cm² in the latanoprost acid 0.01%, 0.05%, and 0.1% arms, respectively). No significant between-arm differences were detected. Secondary and exploratory trichoscopic analyses showed reductions in yellow-dot counts, a decrease in single-hair FUs, and an increase in triple-hair FUs. Safety was favorable, with no serious adverse events. In mechanistic assays, latanoprost acid triggered rapid, concentration-dependent Ca²⁺ flux, whereas equimolar latanoprost produced delayed signals; neither compound altered EdU incorporation. In this pilot proof-of-concept trial, topical latanoprost acid showed a coherent clinical-trichoscopic bioactivity signal, supported by FP receptor-linked signaling in HHDPCs. These findings require confirmation in larger randomized pharmacokinetic/pharmacodynamic-integrated trials designed to optimize dose, confirm efficacy, and further characterize long-term safety. ClinicalTrials.gov, NCT07412587; registered on February 2, 2026.
Hydroxysteroid 11-beta dehydrogenase 1 (11β-HSD1) plays a critical role in metabolic homeostasis by catalyzing the intracellular conversion of cortisone to cortisol. Dysregulated 11β-HSD1 activity is closely associated with metabolic disorders such as type 2 diabetes mellitus, obesity, and glucocorticoid-related inflammation. While small-molecule inhibitors of 11β-HSD1 have shown promise, they primarily suppress enzymatic activity without modulating protein abundance. Here, we report the development of the 11β-HSD1-targeting PROTAC degraders. A series of bifunctional molecules were synthesized based on CRBN- and VHL-recruiting ligands, with AZD8329-derived warheads linked via polyethylene glycol chains. Cellular assays demonstrated efficient, ubiquitin-proteasome-dependent degradation of 11β-HSD1, with H-3-V identified as the most potent degrader. In vivo, H-3-V treatment improved glucose tolerance and enhanced glucose-stimulated insulin secretion in a high-fat diet-induced T2DM mouse model. Molecular dynamics simulations revealed that the H-3-V ternary complex exhibited superior binding energy compared to less active analogs. Collectively, this study introduces a novel chemical modality for 11β-HSD1 modulation and lays the groundwork for future therapeutic development targeting metabolic diseases via protein degradation.
Sigma receptors (SRs), comprising the sigma 1 (S1R) and sigma 2 (S2R) subtypes, represent a distinct class of intracellular proteins that differ fundamentally from classical neurotransmitter receptors. S1Rs act as ligand-operated molecular chaperones primarily localized at mitochondria-associated membranes (MAM) of the endoplasmic reticulum (ER), whereas S2Rs, identified as transmembrane protein 97 (TMEM97), function as modulators of cellular homeostasis. Both subtypes are densely expressed in brain regions critical for emotional and cognitive processing, including the prefrontal cortex (PFC), hippocampus (HIP), amygdala (AMG), and basal ganglia. SRs play a pivotal role in regulating neuronal excitability, synaptic plasticity, and ER stress responses, thereby influencing oxidative stress, synaptogenesis, and neuronal survival. Moreover, their interactions with endogenous neurosteroids and the immune-endocrine system highlight their involvement in complex neuropsychiatric pathophysiology. Consequently, SRs are increasingly recognized as promising targets for pharmacological intervention. Preclinical evidence indicates that S1R agonists exert antidepressant, anxiolytic, and pro-cognitive effects, whereas S1R antagonism or genetic deletion induces behavioural phenotypes relevant to depression and schizophrenia. S2R modulation, in turn, influences psychostimulant sensitivity and emotional regulation, although its underlying mechanisms remain less well defined. Notably, the clinical efficacy of several widely used antidepressants (e.g., fluvoxamine) and antipsychotics have been increasingly attributed to their significant affinity for SRs, suggesting that SR engagement may contribute to their therapeutic profiles. This narrative review synthesizes core molecular and neurophysiological findings and critically evaluates the potential of SR-targeting compounds as next-generation pharmacological strategies for treatment-resistant depression, psychotic disorders, and addiction. Given the continuous emergence of new research, this work provides an updated overview of the current state of knowledge for both researchers and clinicians.
Aminophylline, a bronchodilator used for treating airway obstruction, has been predicted through in silico models to exert antidepressant effects. However, there are still no studies that have validated these claims in a biological system. In this paper, we evaluated the antidepressant effects of aminophylline in mice subjected to chronic restraint stress (CRS). CRS was conducted for a duration of 15 days, with 4-hour daily stress exposure. Aminophylline (5 mg/kg, 10 mg/kg, and 20 mg/kg) and fluoxetine (10 mg/kg, positive control) were administered daily via intraperitoneal injection. Behavioral assessments, including the tail suspension test (TST), forced swimming test (FST), and sucrose splash test (SST), were conducted on days 0, 5, 10, and 15. Molecular docking and pathway analyses were performed to provide insight into its possible mechanism of action. CRS exposure successfully induced depressive-like behavior, characterized by prolonged immobility in the TST and FST and diminished grooming activity in the SST. Administration of aminophylline attenuated these behavioral deficits, reducing immobility time and increasing grooming time in a dose-dependent manner. Molecular docking analysis demonstrated favorable binding of aminophylline to phosphodiesterase 3, phosphodiesterase 4, and the serotonin transporter, targets associated with antidepressant activity. Pathway analysis revealed upregulation of PPAR signaling, calcium signaling, and the synaptic vesicle cycle, while downregulating the glutamatergic synapse pathway. The study provides the first evidence of the antidepressant activity of aminophylline in a validated model of depression, prompting further clinical investigation into its therapeutic potential. Moreover, molecular and biochemical analyses are warranted to validate its precise mechanism of action. [Image: see text]
BACKGROUND: The isolation of intact cell populations from archived tissue is critical for understanding cell-type-specific functions in neuroscience. This study presents a streamlined, bench-ready fluorescence-activated cell sorting (FACS) protocol for the isolation of neuronal and non-neuronal populations from frozen adult rat cortex, specifically optimized for integrated RNA and protein analyses. METHODS: The method employs a gentle enzymatic digestion and mechanical trituration, followed by fixation with zinc salts, which successfully addresses the challenges of dissociating adult brain tissue while preserving key cellular markers. The protocol’s versatility is demonstrated by its compatibility with two distinct collection buffers, enabling the simultaneous preparation of samples for both RNA and protein analysis, performed by qRT-PCR and mass spectrometry, respectively. RESULTS: Neuronal (PE+) and non-neuronal (PE-) fractions were isolated with high purity, as confirmed by significant enrichment of Rbfox3, Map2, and Tubb3 (neuronal) and Gfap, Iba1, Olig2, and Cldn5 (non-neuronal) via qRT-PCR. Subsequent proteomic analysis further confirmed the distinct identities of the sorted populations, with key neuronal and non-neuronal markers showing significant enrichment in their respective fractions. CONCLUSIONS: Despite a conservative yield (ca. 20,000 neurons per sample) prioritized for cellular intactness, the obtained material was sufficient for high-sensitivity qPCR (mRNA/miRNA) and mass spectrometry. This accessible and time-efficient protocol provides a robust platform for parallel RNA and protein analysis of defined brain cell populations from archived frozen tissue.
On December 1st, 2023, the Pharmacovigilance Risk Assessment Committee (PRAC) published a safety warning against using pseudoephedrine in patients with severe or uncontrolled hypertension and severe acute or chronic kidney disease for the risk of developing posterior reversible encephalopathy syndrome (PRES) or reversible cerebral vasoconstriction syndrome (RCVS). Based on this consideration and the availability of other decongestants with the same mechanism of action as pseudoephedrine in the European market, we decided to conduct a safety study to evaluate cases of RCVS or PRES reported with other decongestants. A European, retrospective, post-marketing surveillance study was conducted to describe individual case safety reports (ICSRs) of RCVS and PRES reported with decongestants and to compare the likelihood of reporting these events among decongestants. Data were retrieved from the EudraVigilance database from January 1st, 2001, to December 31st, 2025. A total of 65 ICSRs reported PRES or RCVS with decongestants, most of which referred to female patients aged 18-64 years. A disproportionate reporting of RCVS was found for xylometazoline (reporting odds ratio, ROR: 3.39; 95% confidence interval, CI: 1.94-5.93) and phenylephrine (ROR: 2.00; 95% CI: 1.11-3.61), and a disproportionate reporting of PRES for ephedrine (ROR: 6.44; 95% CI: 1.80-23.10) and phenylephrine (ROR: 3.47; 95% CI: 1.22-9.88) compared to all other decongestants. Our results suggest that all decongestants should be used with caution, and attention should be paid to signs and symptoms of PRES and RCVS when taking these medicines.
Post-traumatic stress disorder (PTSD) is associated with dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis and the sympatho-adrenomedullary system, leading to immune imbalance and alterations in microbiota composition. Stress-induced disruption of intestinal barrier integrity may promote bacterial translocation to peripheral organs, including the spleen. Aripiprazole (ARI), an atypical antipsychotic proposed for PTSD treatment, also modulates the immune system and microbiota. This study investigated the effects of ARI on splenic microbiota composition and splenic neuroendocrine and immune responses in an animal model of PTSD. Rats were exposed to the single prolonged stress (SPS) paradigm to induce a PTSD-like phenotype and treated intraperitoneally with vehicle or ARI for 28 days. Anxiety-like behavior was assessed using the elevated plus maze. Splenic microbiota and gene expression were quantified by real-time PCR in isolated splenocytes following ex vivo stimulation with lipopolysaccharide or phorbol 12-myristate 13-acetate (PMA)/ionomycin. SPS and SPS + ARI reduced the abundance of the phylum Bacteroidetes. SPS increased γ/δ-Proteobacteria and Lactobacillus abundance, effects attenuated by ARI. The presence of specific splenic bacteria correlated with anxiety-like behavior. While lipopolysaccharide-induced responses were unaffected, splenocytes from SPS-exposed rats exhibited increased expression of Th1- and Th17-related genes after PMA/ionomycin stimulation; this effect was reversed by ARI. ARI modulates SPS-induced alterations in splenic microbiota and attenuates heightened Th1- and Th17-associated responses, thereby contributing to the restoration of immune balance. Our findings underscore involvement of the gut-spleen-brain axis in PTSD pathogenesis and immunosuppressive/ microbiota-modulating effects of aripiprazole on splenic immune cells.
Charcot neuroarthropathy (CN) is a debilitating joint disorder that predominantly affects patients with neuropathy, particularly those with diabetic peripheral neuropathy (DPN). CN causes painless, rapid joint destruction and often leads to foot deformity, ulceration, osteomyelitis, and, in severe cases, amputation. Its pathogenesis involves repetitive microtrauma due to loss of protective sensation, triggering an inflammatory cascade that activates osteoclasts (OCs) disproportionately relative to osteoblasts (OBs) via the RANKL-RANK-OPG pathway, resulting in progressive bone loss and joint destruction. This systematic review and meta-analysis evaluated the efficacy of anti-resorptive agents in promoting bone remodeling and alleviating clinical symptoms in patients with active or stable CN. Following Cochrane Collaboration guidelines, we searched MEDLINE, EMBASE, and the Cochrane Library for randomized controlled trials (RCTs) comparing anti-resorptive agents, such as bisphosphonates, denosumab, calcitonin, and parathyroid hormone analogues, with placebo or no treatment in patients with Charcot neuroarthropathy. Two independent reviewers performed data extraction and risk-of-bias assessment using the Cochrane RoB 2 tool. Primary outcomes were bone mineral density (BMD), bone turnover markers (BTMs), time to remission, change in foot temperature, and adverse events. Statistical analyses were conducted using Stata 18, with random-effects models used to pool results. We identified 936 records and nine reports describing seven RCTs met the inclusion criteria. The meta-analysis showed no significant difference in BMD between anti-resorptive agents and control groups. However, anti-resorptive therapy significantly reduced bone resorption markers. Clinical outcomes, including foot temperature change and time to remission, did not differ significantly between groups. Adverse events were similar between the intervention and control groups. Although anti-resorptive agents reduce bone resorption markers in patients with Charcot neuroarthropathy, current evidence does not support their efficacy in improving BMD or providing clinically meaningful symptom relief beyond standard offloading. High-quality clinical trials and mechanistic studies are needed to define the role of these agents in CN management.