Temozolomide is the gold standard chemotherapeutic agent for glioblastoma multiforme. Yet its pharmacological use has been linked to the emergence of depressive- and/or anxiety-like behaviors, probably through the inhibition of hippocampal neurogenesis. Since prior studies reporting these negative effects were based on prolonged treatment paradigms (from 2 weeks to up to 6 months), and given the lack of studies including females, our approach aimed at further characterizing the behavioral effects induced by temozolomide (25 mg/kg, 1 or 2 cycles, 5 days/cycle) in a adult rats of both sexes. Rats were scored across time through specific behavioral tests that capture diverse manifestations of affective-like responses (forced-swim, open field, novelty-suppressed feeding, sucrose preference) or cognitive performance (Barnes maze). At the neurochemical level, we ascertained the effects of 2 cycles of temozolomide on an early stage of hippocampal neurogenesis (neural progenitors, NeuroD) and other potential neuroplasticity targets (mature BDNF, FADD). Temozolomide induced signs of antidepressant-like responses as measured in the forced-swim test in a treatment-duration manner. It also decreased NeuroD and hippocampal FADD, a neuroplastic marker previously associated with the acute and repeated actions of most antidepressants. These results suggested that other mechanisms of action might be associated with temozolomide’s behavioral effects besides hippocampal neurogenesis, such as the correlative one described through the neuroplastic molecule FADD. In conjunction with the prior data, our results suggested cycle- and/or length-dependent treatment effects in terms of temozolomide’s antidepressant- vs. depressant-like profile, while proposing a novel biomarker related to treatment response.
Poly(ADP-ribose) polymerase (PARP) inhibitor maintenance therapy is a standard treatment in selected ovarian cancer patients. However, hematological toxicity remains a clinical problem. The study aimed to identify predictors of early hematologic toxicity during PARP inhibitor maintenance therapy in high-grade serous ovarian cancer. We retrospectively analyzed 130 patients receiving PARP inhibitors (niraparib, 49; olaparib, 81). Blood counts from first-line platinum chemotherapy and from the first 3 months of PARP inhibitor therapy were assessed. Adverse events were graded using the Common Terminology Criteria for Adverse Events v5.0. Weekly changes and minimal values for neutrophils, hemoglobin, and platelets were calculated (excluding post-intervention values). Thrombocytopenia was mainly related to niraparib treatment, and anemia was mainly related to olaparib treatment. During niraparib therapy, higher human epididymis protein 4 (He4) at diagnosis correlated with greater platelet decline and lower platelet nadir. In multivariable linear regression, higher body weight (β = 1.86, 95
Gastrointestinal (GI) tract diseases cause symptoms that significantly affect the quality of life. A promising direction in the search for novel therapeutic options in this field is the investigation of G protein-coupled receptors (GPCRs), whose activity is modulated by their regulator proteins (RGS). In this study, we examined RGS6 involvement in GI inflammation and functional disorders and its effect on cannabinoid (CB), opioid, and serotonin receptor (5HTR)-targeting compounds, Using quantitative PCR, western blot, and ELISA assays, we characterized RGS6 expression in the mouse GI tract and measured GPCR-related secondary messenger expression upon stimulation with GPCR agonists in vitro in Caco-2 cells, both wild type (WT) and RGS6 knock-out (KO). Then, in vivo in a dextran sulfate sodium (DSS) mouse model of colitis using WT, global and tissue-specific RGS6 KO mice, inflammation, antinociceptive effects, and GI motility were examined. In Caco-2 cells, we found that RGS6 KO increased extracellular signal-regulated kinase phosphorylation, which was blocked by CB agonist WIN 55,212-2. Moreover, incubation with WIN 55,212-2 significantly reduced cyclic adenosine monophosphate (cAMP) levels in RGS6 KO but not WT cells. In DSS-induced, acute and chronic-relapsing mouse models of colitis, RGS6 KO resulted in resistance to stress-induced GI hypermotility. WIN 55,212-2 (1 mg/kg ip) substantially prolonged GI transit time and displayed antinociceptive properties that were stronger in the absence of RGS6. Epithelial -specific RGS6 KO did not affect the action of WIN 55,212-2. RGS6 exerts crucial effects on GI physiology by acting on CB signaling in the enteric and/or central nervous system.
The long non-coding RNA - microRNA (lncRNA–miRNA) regulatory axis is a key modulator of immune and inflammatory pathways, and growing evidence supports its contribution to therapeutic response variability. In our previous study, we identified the lncRNA MEG3 as potentially involved in the response to biological drugs, specifically TNFα- and IL17A-inhibitors, in psoriatic arthritis (PsA). To further characterize its role, we performed a bioinformatic analysis to identify MEG3-targeted miRNAs, followed by an exploratory expression profiling analysis using qRT-PCR in a cohort of 54 PsA patients at baseline (T0) and after 12 months of therapy (T12), compared with 15 healthy controls (CTRLs). We identified four candidate miRNA targets: hsa-miR-21-5p, hsa-miR-19b-3p, hsa-miR-19a-3p, and hsa-miR-17-5p. Among these, hsa-miR-17-5p was significantly upregulated in PsA patients at T0 versus CTRLs. Most importantly, we found a significant decrease in hsa-miR-17-5p and hsa-miR-19b-3p levels in Responder patients at 12-month follow-up. We then explored whether MEG3 genetic variability contributes to miRNA modulation. First, we observed that the MEG3 rs941576 genetic variant appears to influence the expression levels of hsa-miR-19a-3p and hsa-miR-19b-3p. Next, in silico analyses indicated that this variant lies within an immune-active enhancer, altering the binding affinity for the transcription factors HIF1A/ARNT2, and suggested a link between MEG3 enhancer activity and hypoxia-responsive regulation of these miRNAs. Lastly, pathway enrichment analysis highlighted that both hsa-miR-17-5p and hsa-miR-19b-3p converge on the TGF-β signalling pathway. Our findings suggest the involvement of a specific MEG3-miRNA network in modulating the response to biological therapies in PsA.
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.
Therapeutic drug monitoring (TDM) may optimize dosing of mycophenolate mofetil (MMF) in patients with nephrotic syndrome, but it is not routinely implemented. When performed, TDM typically relies on total mycophenolic acid (MPA) concentrations, although only unbound MPA is pharmacologically active. This study aimed to develop a population pharmacokinetic (PK) model characterizing the PK and pharmacodynamics (PD) of total and unbound MPA in plasma and saliva of paediatric nephrotic syndrome patients following oral MMF. PK/PD data from 62 children (47 full and 21 limited sampling curves) were analysed using nonlinear mixed-effects modelling. A two-compartment model with first-order absorption best describes unbound MPA PK. Protein binding was captured using a nonlinear saturable binding equation. The final population PK parameter estimates (for unbound MPA) were absorption rate constant (ka), 1.81 h−1 (relative standard error (RSE): 8
Polypharmacology is dedicated to the development of compounds acting on at least two targets (multi-target-directed ligands, MTDLs). In 2025, the European Medicines Agency (EMA) approved 38 drugs, and 11 out of them were MTDLs. Most of them are antibody-drug conjugates, bispecific antibodies, or kinase inhibitors, all of which are indicated for tumor treatment, including datopotamab deruxtecan (hormone receptor-positive, HER2-negative breast cancer), tisotumab vedotin (advanced cervical carcinoma), linvoseltamab (fourth-line treatment of multiple myeloma), and erdafitinib (advanced urothelial carcinoma). The small molecule tiratricol is an orphan drug, which is indicated for the treatment of the very rare Allan-Herndon-Dudley syndrome. The second part of the present review is dedicated to the post-marketing safety surveillance of MTDLs approved by the EMA in 2022–2024. For 19 out of the 27 MTDLs, which are still available on the European market, comprehensive pharmacovigilance studies, mainly based on the Food and Drug Administration (FDA) Adverse Event Reporting System (FAERS), were found. New safety signals have been identified, including Stevens-Johnson syndrome and progressive multifocal leukoencephalopathy. The analysis also revealed a more favorable safety profile of the MTDL tirzepatide (a dual glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide analogue) compared to the single-targeted drug semaglutide (glucagon-like peptide-1 analogue), including lower reporting rates of acute kidney injury and no significant suicidality signal.
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.
Postherpetic neuralgia (PHN) is a chronic neuropathic pain condition that develops following herpes zoster, yet the molecular mechanisms underlying the transition from acute herpetic pain to chronic pain remain unclear. Brain-derived neurotrophic factor (BDNF)–TrkB signaling is implicated in central sensitization, but its role in virus-induced pain chronification is unknown. Therefore, this study investigated the temporal involvement of BDNF–TrkB signaling in acute herpetic pain and in the development of PHN-like pain. A herpes simplex virus type-1 (HSV-1)–induced mouse model of herpetic pain was used to evaluate temporal changes in neurotrophin and Trk receptor expression in the dorsal root ganglia (DRG) and spinal dorsal horn. Pharmacological interventions included intrathecal administration of anti-BDNF, anti-Trk receptor antibodies or TrkB-Fc chimera, and repeated systemic treatment with the TrkB antagonist ANA-12 during the acute phase. Mechanical allodynia was behaviorally assessed. BDNF mRNA expression was significantly upregulated in ipsilateral DRG and spinal dorsal horn during the acute phase, whereas neurotrophin-3 remained unchanged and nerve growth factor showed only transient increases in DRG. No neurotrophin or Trk receptor changes were observed in established PHN-like pain. Intrathecal administration of anti-BDNF antibody, anti-TrkB antibody, or TrkB-Fc chimera significantly attenuated acute mechanical allodynia, whereas anti-TrkA and anti-TrkC antibodies showed no significant effects. None of these interventions affected established PHN-like pain. Repeated administration of the TrkB antagonist ANA-12 during the acute phase attenuated acute herpetic pain and significantly reduced the subsequent incidence of PHN-like pain. BDNF–TrkB signaling plays a phase-specific role in herpetic pain, driving acute pain expression and the transition to chronic PHN-like pain but not pain maintenance. Early TrkB inhibition may represent a disease-modifying strategy to prevent viral pain chronification.
Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is an inhibitory receptor expressed by multiple tumors, including colorectal cancer (CRC). CEACAM1 exists as a long isoform (L) containing immunoreceptor tyrosine-based inhibitory motifs (ITIMs) and a short isoform (S) lacking ITIMs. While microsatellite instability (MSI) CRCs respond to immunotherapy, microsatellite-stable (MSS) CRCs remain resistant, highlighting the need for alternative therapeutic targets. We previously developed an agonistic anti-CEACAM1 monoclonal antibody (mAb), CCM5.01, and demonstrated its CEACAM1-dependent inhibitory effects in melanoma. Here, we evaluated CCM5.01 inhibitory activity on CRC cell lines and related it to L/S ratio. CRC cell lines HT-29, LOVO, and HCT-116 were treated with CCM5.01 and analyzed by proliferation assays before and after transfection with L or S CEACAM1 isoforms. Transfected cells treated with CCM5.01 were analyzed for signaling and in 3D spheroid models. CEACAM1-positive CRC cells showed dose-dependent inhibition in response to CCM5.01 treatment and were associated with L/S isoform expression. High CEACAM1-L expression induced growth inhibition and apoptosis, while CEACAM1-S promoted cellular activity. Similar isoform-dependent effects were observed in spheroid models. CEACAM1-mediated signaling in CRCs critically depends on the L/S isoform ratio, supporting CEACAM1-targeted activating therapy as a potential strategy in both MSI and immunotherapy-resistant MSS CRC.
Serotonin (5-hydroxytryptamine; 5-HT) is a key neuromodulator involved in the regulation of mood, appetite, aggression, and impulse control. Dysregulation of central 5-HT function has been implicated in alcohol use disorder (AUD) and comorbid depression. This review summarizes clinical and preclinical evidence on the role of 5-HT in AUD development, heterogeneity, and treatment response. Alterations in 5-HT function may be shaped by stress and genetic variation in serotonergic genes, including TPH2 and SLC6A4, contributing to individual vulnerability to AUD. Reduced 5-HT activity increases the risk of developing AUD, particularly Cloninger’s type II, characterized by early onset, violent, and antisocial behaviors. Consistently, Tph2-deficient mice, which lack central 5-HT, exhibit increased ethanol consumption and behavioral features resembling Cloninger’s type II alcohol dependence. Similarly, alcohol-preferring rat lines show reduced 5-HT levels, decreased serotonergic projections to the cortex, and reduced prefrontal 5-HT2A receptor binding, implicating raphe-prefrontal serotonergic projections as a critical pathway modulating vulnerability to AUD. Given the heterogeneity of AUD, the efficacy of selective 5-HT reuptake inhibitors (SSRIs) remains limited, with beneficial effects observed only in less severe, later-onset forms. Recently, serotonergic psychedelic-assisted therapies have attracted considerable interest as potential AUD treatments; mechanistically, their effects may be linked to activation of 5-HT2A receptors in the prefrontal cortex – a brain region known to be dysfunctional in AUD. The reviewed literature highlights the need for improved stratification of AUD subtypes and validation of 5-HT-related biomarkers to guide personalized therapeutic approaches.
BackgroundThe aryl hydrocarbon receptor (AHR) plays a key role in immune regulation and drug metabolism, potentially influencing methotrexate (MTX) treatment outcomes in patients with rheumatoid arthritis (RA). This exploratory study investigated the relationship between AHR activity and MTX responsiveness, and examined whether combination therapy with tocilizumab (TCZ), an interleukin (IL)-6 receptor inhibitor, could influence MTX resistance and treatment response.MethodsWe employed in silico docking to assess MTX binding to the AHR Per-Arnt-Sim (PAS)-B domain. Ex vivo and in vitro models using peripheral blood mononuclear cells (PBMCs) from RA patients and healthy donors were also used. Flow cytometry was used to analyze AHR expression across immune cell subtypes. Additionally, HepG2 cells served as a pharmacological model to study the interaction of MTX and TCZ with AHR and the expression of drug transporter genes.ResultsAHR expression was significantly higher in monocytes from good responders to MTX than in those from poor responders and MTX-intolerant patients, suggesting that monocytes were the PBMC subset most strongly associated with AHR-related patterns of MTX response. In silico analysis supported the binding of MTX to the PAS-B domain of AHR. The in vitro model confirmed that monocytes were the most responsive subset in the context of AHR-related changes. Treatment with TCZ tended to reduce the proportion of AHR-positive monocytes, whereas co-treatment with MTX shifted AHR toward a pattern comparable to that in good responders or under control conditions.ConclusionsOur findings underscore the complexity of MTX pharmacodynamics and highlight AHR as a potential biomarker for predicting treatment response in RA patients. The combination of MTX and TCZ modulated AHR activity and could inform personalized therapeutic strategies, especially in patients exhibiting MTX resistance or intolerance. While preliminary, this multi-layered investigation-combining patient samples, 3D cultures, and molecular docking-supports further research into AHR-modulating therapies in RA.Clinical trial numberNot applicable.
Hypercholesterolemia is a major risk factor that contributes to the development and/or aggravation of cardiovascular diseases. High cholesterol levels are frequently managed with hypocholesterolemic agents, either synthetic or natural. Beyond their cholesterol-lowering effects, these compounds can also affect the host’s gut microbiota. This review examined the extent and quality of the modulatory activity of selected hypocholesterolemic agents (n = 11) on the gut microbiota profile, based on a comprehensive literature search and analysis of interactions between the gut microbiota and distinct cholesterol-lowering compounds. With a few exceptions, anti-cholesterol interventions were associated with changes in gut microbiota β-diversity and a decrease in the Bacillota/Bacteroidota ratio. Further analyses at family and genus levels revealed patterns of modulation that clustered by molecular target and bioavailability. Overall, gut microbiota shifts favoured short-chain fatty acid (SCFA)-producing bacteria over inflammation-promoting taxa. Furthermore, for some anti-cholesterol compounds, microbial metabolism and the concomitant release of bioactive metabolites suggested partially microbiota-dependent cholesterol-lowering effects. These findings may inform new strategies for cholesterol management from a gut-health perspective; however, further research is needed to establish causality and draw robust conclusions regarding medication–microbiota relationships.
Background Oropharyngeal adverse events (O-AEs) represent a potential safety concern associated with several drugs and/or vaccines. Although often underestimated, these events may provide valuable insights into a patient's overall clinical condition, appearing initially mild but later worsening. Therefore, this study aimed to analyze O-AEs related to drugs and/or vaccines using structured safety data. Methods Safety reports from three Italian regions were retrieved from the national pharmacovigilance database (rete nazionale di farmacovigilanza, RNF) and analyzed for the period 2022-2024. All reports were structured and analyzed according to the International Council of Harmonisation (ICH) E2B (R3) format. Additionally, reporting odds ratios (RORs) were calculated to compare the likelihood of O-AEs being reported by different categories of reporters (e.g., physicians, other healthcare professionals, or patients). Results Over three years, 47,664 reports were collected, of which 1,740 (3.6%) contained at least one suspected O-AE. Most patients were female (65.7%) with a median age of 55 years (IQR 38-66). The majority of reports described non-serious events (66.7%), and outcomes were favorable in most cases (75.9%). The safety reports related to drugs (90.5%) were largely more than those related to vaccines (9.5%). A total of 129 cases of medication-related osteonecrosis of the jaw (MRONJ) were identified, most of which were serious and had unfavorable outcomes. Disproportionality analysis revealed that physicians were less likely to report O-AEs than patients (ROR=0.72; 0.62-0.84; P<<0.05) and more likely than nurses (ROR=1.34; 0.98-1.87; P<0.05). Compared with 2019-2021, the main difference was the lower proportion of vaccine-related reports, which declined from 47.8% to 9.5% in 2022-2024. Conclusions Only a small proportion of safety reports involved O-AEs. These findings highlight the importance of enhancing awareness among physicians (particularly dentists) regarding O-AEs, and of fostering a collaborative pharmacovigilance culture across healthcare providers, thereby improving patient safety through more timely and reliable reporting.
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.
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.
Gut microbial β-glucuronidase (GUS) regulates the deconjugation of glucuronidated compounds, thereby influencing the enterohepatic circulation of xenobiotics and endogenous metabolites. Although many orally administered drugs reach the intestinal lumen, their direct effects on microbial enzyme functions remain incompletely understood. In this study, we systematically examined the impact of ten commonly prescribed central nervous system (CNS)-active drugs on microbial GUS activity. GUS inhibition was evaluated using purified GUS from Escherichia coli (E. coli), E. coli cell lysates, and intact bacteria, with p-nitrophenyl β-D-glucuronide as the substrate. Intracellular drug accumulation was quantified by high-performance liquid chromatography-tandem mass spectrometry, bacterial growth was assessed by optical density, and GUS activity in mouse cecal contents was analyzed ex vivo. Molecular docking and molecular dynamics simulations were conducted to characterize drug–GUS interactions. Purified GUS screening identified aripiprazole (ARI) and duloxetine hydrochloride (DLX) as inhibitors. In intact E. coli, ARI, but not DLX, suppressed intracellular GUS activity without affecting bacterial growth. ARI also accumulated in E. coli at higher levels than DLX. In ex vivo assays, ARI showed inhibitory effects on GUS activity. Computational analyses suggested that ARI and DLX may preferentially interact with distinct regions of GUS, with ARI exhibiting more favorable binding energetics. These findings suggest that specific CNS-active drugs may directly modulate gut microbial GUS activity in E. coli under experimental conditions. In particular, ARI inhibited intracellular GUS activity, raising the possibility that certain neuropsychiatric drugs may influence microbial metabolic functions in addition to their canonical pharmacological targets.
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.
Neuropathic pain remains a significant clinical problem that necessitates the development of more effective treatment options. Compared with commonly used clinical drugs, such as gabapentin and pregabalin, mirogabalin is a novel gabapentinoid that exhibits greater affinity and selectivity for the α2δ-1/-2 subunits of voltage-gated calcium channels, properties associated with strong nociceptive modulation. The aim of this study was to determine whether and how mirogabalin influences neuropathy symptoms and the antinociceptive effects of antidepressants (amitriptyline and duloxetine). Studies were conducted in mice after chronic constriction injury of the sciatic nerve. On Day 7, a single intraperitoneal administration of mirogabalin (5–60 mg/kg), amitriptyline (1–10 mg/kg), or duloxetine (10–30 mg/kg) at various doses was performed. Additionally, mirogabalin (10 mg/kg) was chronically injected twice daily, while amitriptyline (5 mg/kg) or duloxetine (10 mg/kg) was administered once daily. Hypersensitivity was assessed using the von Frey and the cold plate tests. Using a mouse model of neuropathy, a single injection of mirogabalin, amitriptyline, or duloxetine reduced tactile and thermal hypersensitivity to a similar degree, with the most durable effect observed with mirogabalin, persisting up to 6 hours. Repeated twice-daily intraperitoneal administration of mirogabalin combined with once-daily intraperitoneal dosing of amitriptyline or duloxetine resulted in a potentiated anti-nociceptive response in the used model. These results may serve as a basis for further evaluation of the potential use of mirogabalin in the clinical treatment of neuropathic pain. Furthermore, they suggest that using mirogabalin together with amitriptyline or duloxetine may provide significant therapeutic benefits, enabling the use of lower doses of these medications, which may reduce the risk of adverse events.
Cancer is a complex and multifactorial disease, often characterized by disruptions in key cellular processes such as gene expression and DNA topology. Histone deacetylases (HDACs) and topoisomerases (Topo) are two major molecular targets for cancer therapeutics due to their key role in maintaining DNA topology and contributing to cancer development. HDACs modify chromatin accessibility, while Topos resolve DNA supercoiling during replication. Targeting these pathways individually has shown therapeutic potential; however, recent advancements emphasize combination therapies and dual inhibitors as promising strategies to enhance anticancer efficacy. Combination therapies involving HDAC and Topo inhibitors leverage their complementary mechanisms of action to induce synergistic effects, leading to improved tumor suppression. Similarly, dual inhibitors, which integrate the functionalities of HDAC and Topo inhibitors into a single molecule, offer the potential to streamline treatment regimens, overcome drug resistance, and minimize adverse effects. This review provides a comprehensive overview of the therapeutic potential of HDAC-Topo combination therapies and dual inhibitors, emphasizing their mechanistic synergy, pharmacological benefits, and associated clinical challenges. It highlights the need for continued research to address limitations and improve their effectiveness and safety for broader applications in oncology.Clinical trial numberNot applicable.
