Background/Objectives: Non-medical use of GHB poses a public health risk due to the risk of overdose. GHB exhibits nonlinear toxicokinetics mediated by monocarboxylate transporters (MCTs), and previous studies have demonstrated sex-related differences in transporter regulation and GHB toxicokinetics. This study investigated the impact of exogenous estrogen and progesterone on GHB toxicokinetics and toxicodynamics, and evaluated the efficacy of AR-C155858 (an MCT1 inhibitor) to treat GHB overdose. Methods: Ovariectomized (OVX) females and castrated (CST) male Sprague Dawley rats were treated with estrogen and/or progesterone or a corresponding placebo for 21 days. Rats received intravenous GHB (1000 or 1500 mg/kg) or GHB + AR-C155858 (1500 mg/kg + 1 mg/kg), and plasma and urine samples were collected for 8 h post-dose. GHB concentrations were quantified by a validated LC/MS assay. Results: Female sex hormones significantly affected GHB toxicokinetics. Treatment with estrogen and progesterone (alone or in combination) reduced GHB exposure in OVX and CST rats through altered GHB clearance. Alterations in renal and metabolic clearance were dose-dependent. GHB exposure was significantly decreased with the co-administration of AR-C155858, as a result of increased GHB renal clearance. Conclusions: Female sex hormones influence GHB toxicokinetics, with combined estrogen/progesterone treatment producing the greatest reductions in systemic GHB exposure and toxicity. Monocarboxylate transporter inhibition with AR-C155858 significantly decreased GHB exposure and sedative effect, supporting its potential as a therapeutic strategy for GHB overdose across diverse populations.
Background: Due to drug efficacy, adverse reactions, and individual differences, clinicians face significant challenges in drug selection for the treatment of depression. However, the study on predicting antidepressant treatment response based on pharmacogenomics has not yet been determined. Methods: A total of 178 participants were recruited and randomly assigned to either pharmacogenomics-guided antidepressant treatment or regular antidepressant treatment. All participants completed pharmacogenomic sampling at baseline and underwent neuropsychological systemic assessments at baseline and at 4-week, 8-week, 16-week, and 32-week follow-ups. Results: Compared with the control group, the pharmacogenomics-guided group showed significant differences in the effect of DARS scores within 32 weeks (PBonferroni < 0.05), showing a significant time and group effect (p < 0.001; p = 0.007, respectively). In addition, the pharmacogenomic-guided group showed significant differences in ASEC scores within 32 weeks (PBonferroni < 0.05), showing a significant time effect, group effect, and group × time effect (p < 0.001; p = 0.018; p = 0.009, respectively). However, there was no significant difference in HAM-D17, HAM-A, PDQ-D, and PSQI between the two groups within 32 weeks (PBonferroni > 0.05). Conclusions: Pharmacogenomics-guided antidepressant prescribing effectively alleviates anhedonia and reduces adverse effects in patients with first-episode depression, but there is no significant improvement in major psychopathological symptoms. These findings carry important implications for the early application of pharmacogenomic testing in primary care settings for depression.
Background/Objectives: Duloxetine is a serotonin–norepinephrine reuptake inhibitor used for psychiatric and chronic pain conditions. It enhances serotonergic and noradrenergic neurotransmission by inhibiting the reuptake of serotonin and norepinephrine. Acute poisoning, most commonly resulting from intentional oral ingestion, may frequently present with neurological and cardiovascular manifestations. Although duloxetine is widely prescribed, evidence regarding acute overdose remains limited, and no validated toxic duloxetine concentration threshold reliably predicts the severity of duloxetine toxicity. This systematic review aimed to synthesize published cases of duloxetine poisoning and characterize clinical manifestations, management strategies, and outcomes. Methods: We searched PubMed and Google Scholar for case reports and case series describing cases of duloxetine poisoning. We included cases of acute duloxetine poisoning with a clearly toxic ingested dose and/or analytically confirmed toxic duloxetine concentrations. Results: A total of 18 publications describing 23 patients were included in this systematic review. Duloxetine intoxication was classified as suicidal in 17 cases (73.9%), accidental in 5 cases (21.7%), and not specified in one case (4.3%). In most cases (78.3%), duloxetine overdoses involved co-ingestion of other central nervous system depressants. The most common clinical manifestation was altered mental status, ranging from somnolence, confusion, disorientation, and drowsiness to impaired consciousness, coma, or unresponsiveness in severe cases. Nearly half of the included duloxetine poisoning cases (47.8%) were fatal. Conclusions: In conclusion, acute duloxetine intoxication is most frequently associated with the co-ingestion of other central nervous system depressants, and early recognition with prompt supportive management is essential to minimize the risk of fatal outcomes.
Background/Objectives: The pharmacological management of inflammation remains a clinical challenge, driving the demand for advanced topical formulations. This study utilizes a “smart-NADES” (natural deep eutectic solvent) paradigm to develop an innovative anti-inflammatory topical system. The aim was to evaluate the integrated antioxidant potential, alongside the in vitro and in vivo anti-inflammatory activities, of a novel smart-NADES licorice root extract and its hydrogel formulation. Methods: A NADES system composed of D-sorbitol and L-lactic acid (3:1) was employed for licorice root extraction. The antioxidant capacity was assessed using three independent assays and integrated via the Relative Antioxidant Capacity Index (RACI), while phytochemical interactions were quantified using the Chou–Talalai Combination Index (CI). In vitro anti-inflammatory activity was evaluated via protein stabilization capacity, and in vivo efficacy was validated using a formalin-induced paw edema model in mice. Results: The NADES extract contained glycyrrhizic acid levels of 6.3 ± 0.3 mg/g and showed strong antioxidant synergism in the DPPH assay (CI = 0.49 ± 0.07). The extract exhibited potent total antioxidant capacity (IC50 = 11.9 ± 0.6 μg/mL) with a superior RACI score (1.23). In vitro protein stabilization (IC50 = 63 ± 5 μg/mL) was comparable to diclofenac sodium. The 5% hydrogel numerically surpassed the commercial 2% diclofenac Emulgel (67.5% vs. 32.9% inhibition, respectively) at 24 h, although the direct pairwise comparison did not reach statistical significance (p = 0.186). Conclusions: The developed smart-NADES licorice hydrogel represents an effective, green formulation with pronounced topical anti-inflammatory properties, establishing a robust pharmacological rationale for advanced topical drug delivery.
Background/Objectives: In this study, we present an analysis of ayahuasca, a psychedelic preparation containing N,N-dimethyltryptamine (DMT) and β-carbolines, such as harmine (HRM), a reversible monoamine oxidase A (MAO-A) inhibitor that enables the oral bioavailability of DMT. CYP2D6 is a highly polymorphic enzyme associated with interindividual variability in drug exposure, but its influence on the pharmacokinetics of ayahuasca alkaloids remains poorly understood. Methods: Using physiologically based pharmacokinetic (PBPK) modeling, we simulated scenarios for poor (PM), normal (NM), and ultra-rapid (UM) metabolizers by adjusting CYP2D6 enzyme expression for each phenotype. Results: PMs showed increased systemic exposure to DMT (AUC +53.3%; Cmax +40.5%) and HRM (AUC +30.6%; Cmax +22.8%), while UMs exhibited reduced exposure to both compounds. Conclusions: These findings highlight the significant impact of CYP2D6 polymorphisms on the pharmacokinetics of DMT and HRM, reinforcing the value of PBPK modeling for predicting interindividual variability and potential clinical risks.
Background: PRMT5, or protein arginine methyltransferase 5, is recognized as an epigenetic regulator that suppresses gene transcription through symmetric dimethylation of histone arginine residues, including histone H4 arginine 3 (H4R3me2s) and histone H3 arginine 8 (H3R8me2s), modifications associated with chromatin condensation and transcriptional repression. PRMT5-mediated methylation has been associated with recruitment of polycomb repressive complex 2 (PRC2) and enhancer of zeste homolog 2 (EZH2)-mediated H3K27me3 deposition, contributing to stable repression of tumor suppressor genes and apoptosis-related effectors in breast cancer. Methods: The molecular and functional impacts of PRMT5 inhibition were studied in TNBC cell lines with a pharmacological inhibitor (CMP5). Cellular responses were evaluated using a viability assay, qPCR, Western blotting, Annexin V/PI staining, and transwell migration/proliferation assays. Results: PRMT5 inhibition substantially reduced TNBC viability in a time- and dose-dependent manner. EZH2 was downregulated, whereas the tumor suppressor retinoblastoma-like protein 2 (RBL2) was induced, concomitant with low expression of Cyclin D1. These changes were accompanied by upregulation of pro-apoptotic effectors (Caspase-3, Caspase-10, death-associated protein 1 (DAP1), and BCL2-associated x protein (BAX) and repression of the pro-survival B-cell lymphoma 2 (BCL2), consistent with apoptosis-associated molecular responses. Functionally, CMP5 treatment was associated with reduced migratory behavior in TNBC cells under the experimental conditions tested. Conclusions: These findings suggest that PRMT5 inhibition by CMP5 is associated with reduced TNBC cell viability, impaired migration, increased expression of apoptosis-associated regulators and enhanced apoptotic cell death as measured by Annexin V/PI analysis in vitro. Further mechanistic and in vivo studies are required to clarify the therapeutic relevance of PRMT5 inhibition in TNBC.
Background/Objectives: The non-neuronal cardiac cholinergic system (NNCCS) is known to synthesize ACh independently of the parasympathetic nervous system, thereby regulating cardiac homeostasis, which includes sustainability of energy metabolism, anti-inflammatory and anti-ischemic properties, electrical stability, and mitochondrial calcium handling. Given these beneficial functions of NNCCS, we were prompted to search for an inducer. One such inducer is SNPiP, a novel low-molecular-weight chemical compound developed by us. SNPiP accelerates ACh synthesis in the heart via cGMP elevation and, intriguingly, enhances diastolic function, increasing cardiac output and end-systolic pressure without elevating heart rate. However, the pharmacokinetics of SNPiP remain unknown, which led us to conduct the present study. Methods and Results: We found that the half-life of SNPiP in the blood was extremely short, similar to that of a nitric oxide (NO) donor, S-nitroso-N-acetyl-DL-penicillamine. This short half-life is caused by the rapid distribution of SNPiP into organs, including the heart, kidney, and liver. In addition, once transferred into blood cells, SNPiP itself became stable and remained intact for up to 1 h. Moreover, the short half-life was partly explained by the rapid degradation of SNPiP and concomitant loss of the nitroso group in the blood. Notably, when rats were treated with SNPiP, NO levels in the heart elevated bimodally: immediately after administration and again about 12 h later, coinciding with the previous report of NNCCS upregulation and accelerated ACh synthesis with NO production. Importantly, our previous transcriptome analysis of SNPiP-treated hearts supports these findings, as it revealed upregulation of diastolic function-related genes and proteins. Conclusions: Collectively, these results clarify the pharmacokinetics of SNPiP and demonstrate that, despite a shorter half-life, SNPiP is efficiently distributed to the heart, where it confers beneficial effects through induction of NNCCS.
Neurodegenerative diseases (NDs) such as Alzheimer’s, Parkinson’s, and Huntington’s disease are progressive and currently incurable conditions characterized by the deterioration of neuronal structure and function. Its incidence is increasing, primarily driven by global aging, and it represents a significant public health concern. Traditional therapies offer only symptomatic relief and are unable to halt or reverse the underlying neurodegenerative processes. One of the key challenges in developing effective treatments is the presence of biological barriers, such as the blood–brain barrier (BBB), which limits drug delivery to the central nervous system (CNS), namely the brain. Nanotechnology has emerged as a promising tool to overcome these obstacles. Nanoparticles (NPs), due to their small size, biocompatibility, and versatility, can be engineered to cross the BBB, protect therapeutic agents from degradation, and deliver them precisely to target sites in the brain. This work explores the current advances in lipid and polymeric-based nanoparticle (LNPs and PNPs, respectively) drug delivery systems (DDS) and their application in preclinical studies for the treatment of the NDs previously mentioned. The presented studies suggest that this strategy holds great potential, offering new perspectives and emerging strategies to improve therapeutic outcomes for NDs, and promote neuroprotection of the brain.
Background: Prostate cancer remains one of the most prevalent malignancies worldwide, with high mortality in advanced and metastatic stages. Drug repurposing offers a cost-effective and time-efficient strategy to identify new therapeutic options. Objectives: This study aimed to apply physiologically based pharmacokinetic (PBPK) modeling to evaluate repurposed antiretroviral drugs efavirenz (EFV), etravirine (ETV), and saquinavir (SAQ) in prostate cancer, and to assess potential drug–drug interactions (DDIs) between EFV and ETV. Methods: PBPK models for EFV and SAQ were obtained and an ETV was developed and validated using literature and ADMET Predictor® data. Prostate tissue models were modified to simulate malignant conditions, and population-based simulations examined the influence of age and obesity. The GastroPlus® DDI module was applied to explore mechanistic interactions between EFV and ETV under different physiological scenarios. Results: Tumor-specific prostate tissue alterations produced minimal systemic pharmacokinetic changes but increased total drug accumulated in simulated tissue, with differences in unbound concentrations, while demographic variables such as age and weight significantly affected drug exposure, which are comorbidities in prostate cancer. Lighter individuals exhibited higher plasma concentrations across all drugs, consistent with known previously reported pharmacokinetic trends in obese individuals. DDI simulations indicated only minor changes in ETV pharmacokinetics when combined with EFV, with no clinically significant interaction detected. Conclusions: The integration of PBPK modeling, population variability, and DDI analysis highlights the potential of SAQ, EFV, and ETV as viable drugs for prostate cancer repurposing, but with a heavy focus on dosing personalization. In silico approaches provide a useful framework for early preclinical evaluation and the optimization of repurposed drugs, supporting the early evaluation of repurposed drug candidates in oncology.
Wounds, particularly chronic wounds, represent an increasing challenge for global health systems, affecting millions of people worldwide, and are often associated with persistent infections, biofilms, and multidrug-resistant microorganisms (MDRMs). In this context, the search for effective therapeutic alternatives has driven interest in photodynamic therapy (PDT), an approach in which light-excited photosensitizers promote the generation of reactive oxygen species (ROS) with antimicrobial and wound healing properties. Although first- and second-generation organic photosensitizers are widely used, they have significant limitations, including low aqueous solubility, self-aggregation, reduced photostability, and unsatisfactory ROS quantum yields. To overcome these drawbacks, various nanotechnology-based strategies have been explored. Among them, metallic nanoparticles stand out because they serve as carriers and exhibit intrinsic photosensitizing activity, high resistance to photobleaching, and remarkable extinction coefficients, which favor efficient singlet oxygen generation. Furthermore, metals such as gold and silver can enhance the performance of organic photosensitizers through a process known as metal-enhanced singlet oxygen generation, whereas others, such as copper, zinc, manganese, and magnesium, actively participate in biochemical events associated with the inflammatory and regenerative phases of wound healing. Considering these advances, this review compiles evidence published over the past five years regarding the use of metallic or metal-containing nanoparticles in PDT for acute and chronic wounds, with an emphasis on in vivo studies. In addition, we discuss the epidemiological and pathophysiological aspects of wounds and the intrinsic wound healing and antimicrobial properties of metallic compounds, thereby providing an integrated and up-to-date perspective.
Androgens regulate skeletal muscle, bone, erythropoiesis, and male reproductive function via the androgen receptor (AR), a ligand-dependent transcription factor. Pharmacologic modulation of AR has been pursued for clinical and non-medical purposes. Anabolic androgenic steroids (AAS), synthetic testosterone derivatives, act as full AR agonists, broadly activating multiple tissues. While effective in promoting muscle growth and strength, AAS cause well-known adverse effects, including hypothalamic–pituitary–gonadal (HPG) axis suppression, dyslipidemia, hepatotoxicity, cardiovascular disease, tendon injury, and neuropsychiatric disturbances. Selective androgen receptor modulators (SARMs) aim to stimulate AR in muscle and bone while minimizing androgenic effects in prostate and skin. They induce ligand-specific AR conformations, altering coactivator and corepressor recruitment, and avoiding metabolism by 5α-reductase or aromatase. Preclinical studies show favorable anabolic-to-androgenic ratios, but clinical translation is limited. Early human trials report modest lean mass gains, variable functional outcomes, and dose-dependent testosterone suppression. Emerging evidence also suggests cardiotoxicity, tendon injury, and liver toxicity, though long-term effects are unclear. Pharmacokinetically, SARMs have predictable oral absorption and moderate half-lives, enabling once-daily dosing, unlike AAS. This review compares AAS and SARMs in molecular mechanisms, pharmacokinetics, and safety. While SARMs offer partial tissue selectivity and reduced adverse effects, risks remain, and long-term safety is uncertain. Regulatory oversight is limited, and non-medical use is rising. Preclinical and clinical studies are needed to clarify whether SARMs can separate anabolic benefits from androgenic toxicity and inform safe clinical application.
ESKAPE pathogens represent a priority clinical threat due to their multidrug-resistance, persistence in biofilms, and ability to evade antibiotic therapy. In response to these limitations, antimicrobial peptides (AMPs) have emerged as promising platforms for the development of novel anti-infective strategies. This review analyzes the potential of AMPs against ESKAPE pathogens, integrating their main classes, mechanisms of action, design strategies, and barriers to clinical translation. Natural, synthetic, and peptidomimetic AMPs are examined, along with lytic mechanisms, intracellular targets, anti-virulence effects, quorum quenching, and immunomodulation. In addition, in silico design approaches, multi-objective prediction, and molecular optimization strategies—including stereochemical modifications, cyclization, lipidation, PEGylation, and hybrid design—are discussed. Finally, their activity against ESKAPE biofilms is addressed, together with current limitations related to stability, toxicity, delivery, and preclinical validation.
Introduction: Hereditary hemochromatosis is a genetic disorder characterized by dysregulation of iron homeostasis, resulting in excessive intestinal iron absorption and progressive iron deposition in vital organs. The hepcidin–ferroportin axis plays a central role in the pathophysiology of the disease, particularly in cases associated with mutations in the HFE gene. Persistent iron overload may lead to progressive injury in target organs and functional impairment. Methods: A brief description of the pathological basis of hereditary hemochromatosis was conducted together with a systematic review of interventional clinical trials registered on ClinicalTrials.gov. Studies with available results were included regardless of clinical phase or recruitment status. Relevant data regarding therapeutic interventions and iron metabolism parameters were extracted and descriptively analyzed. Results: Three studies met the inclusion criteria: one evaluating the hepcidin mimetic Rusfertide (PTG-300), another assessing synthetic hepcidin LJPC-401, and a third investigating the iron chelator Deferasirox. Discussion: These therapies demonstrated biological activity in modulating iron metabolism parameters; however, none proved superior to therapeutic phlebotomy, which remains the standard treatment, in terms of efficacy. Conclusions: Although emerging pharmacological therapies targeting iron metabolism show promising biological effects, current clinical evidence remains limited. Therapeutic phlebotomy continues to represent the first-line treatment for hereditary hemochromatosis, and further clinical trials are necessary to determine the potential role of these novel therapeutic approaches.
Structure-based strategies are widely used in tuberculosis drug discovery; however, their translational impact remains limited. This review examines how structure-based virtual screening (SBVS) is applied in practice to Mycobacterium tuberculosis targets and explores why docking-derived predictions frequently fail to translate into measurable biological activity. Rather than treating docking scores as quantitative predictors of potency, representative case studies are analyzed to demonstrate that SBVS is most effective when employed as a prioritization framework integrated with appropriate target preparation, physicochemical filtering, and early experimental validation. Across diverse targets, molecular dynamics simulations emerge as a critical discriminator, enabling the identification of binding instability and false-positive hits that persist after static docking. Tuberculosis-specific constraints—including cofactor-dependent catalysis, resistance-associated mutations, membrane-rich environments, and permeability barriers—are discussed as key factors decoupling in silico affinity from whole-cell efficacy. Collectively, these observations support a workflow-oriented view of computational drug discovery in tuberculosis, in which iterative integration of structural modeling and experimental validation is required for meaningful lead identification.
Background: Addictive disorders are highly heterogeneous and frequently comorbid, limiting the clinical utility of categorical diagnoses. Transdiagnostic pharmacology seeks to address these limitations by targeting symptom dimensions and shared neurobiological processes across addictions. Methods: We conducted a theory-driven narrative review of studies indexed in MEDLINE, PubMed, LILACS, and Web of Science (October–November 2025), integrating clinical, mechanistic, and dimensional evidence. Findings were organized using the Dysregulation Phenomena of the Three Main Modes of the Predostatic Mind and the Advanced Cognitive Emotional Regulation Therapy (DREXI3/ACERT) framework, which conceptualizes addiction as dysregulation across three interacting systems—Alarm, Seeking, and Balance—and six transdiagnostic symptom dimensions, with a proposed expansion into twenty clinically observable domains (TDPM-20). Results: Pharmacological interventions consistently target neurobiological systems related to stress, reward, impulsivity, and compulsivity. Across studies, the most clinically relevant outcomes remain abstinence, reduction in substance use, and treatment retention. While these outcomes are essential, expanding outcome frameworks to incorporate dimensional and mechanistically informed measures may enhance the identification of clinically meaningful subgroups. Across studies, multiple pharmacological classes show transdiagnostic potential, but their clinical application remains variably aligned with dimensional clinical profiles. Conclusions: A dimensionally oriented approach grounded in neurobiological principles may improve alignment between clinical processes and therapeutic strategies. The DREXI3/ACERT model provides a structured framework for individualized treatment planning and research integration. This approach should be understood as complementary to, rather than a replacement for, established evidence-based treatments for specific substance use disorders, particularly in contexts where therapeutic options remain limited or insufficient. Advancing transdiagnostic pharmacology will require broader dimensional stratification, expanded outcome frameworks capable of capturing patient heterogeneity, and integrative trial designs to strengthen precision psychiatry in addictive disorders.
Background: This study investigates a novel alginate–gelatin hydrogel incorporating polyphenol-rich grape skin extract as a multifunctional therapeutic system for diabetic wound healing. The extract was obtained by ultrasound-assisted extraction and formulated into a biopolymer hydrogel designed to combine optimal moisture retention with the controlled release of bioactive compounds. Methods: A streptozotocin-induced diabetic rat model was used to evaluate wound contraction, collagen deposition, oxidative stress parameters, and systemic inflammatory markers over a 15-day period. Animals were assigned to four groups: untreated control, silver sulfadiazine (SSD), empty hydrogel (EH), and extract-loaded hydrogel (LH). Results: The LH formulation demonstrated superior wound closure, reaching 97.1% by day 15, significantly outperforming SSD and other groups. Hydroxyproline levels were markedly elevated in LH-treated tissues, indicating enhanced collagen synthesis and extracellular matrix formation. Redox analyses revealed substantial reductions in TBARS and significant increases in SOD, CAT, and GSH, confirming the strong antioxidative activity of the incorporated extract. Moreover, LH treatment produced pronounced decreases in IL-6 and TNF-α, restoring inflammatory balance and facilitating timely progression from the inflammatory to proliferative phase. Conclusions: These effects are attributed to the synergistic actions of grape skin polyphenols which exerted broad biochemical and structural benefits essential for diabetic wound repair. Overall, this sustainable, bioactive hydrogel represents a promising alternative for advanced wound care.
Treatment for large critical-sized bone defects and impaired fracture healing remain challenging. Clinically used protein-based osteoinductive factors, such as recombinant bone morphogenetic proteins (BMPs), can be effective; however, they are costly and limited by stability, dose-delivery issues, and safety concerns. Preclinical small molecules offer an alternative because they are chemically stable, scalable to manufacture, and readily integrated for systemic administration or localized release from scaffolds, hydrogels, cements, and implant coatings. With an emphasis on delivery formats and mechanistic themes, this review examines small molecules that have been shown to improve bone regeneration in preclinical models, contrasting those of biological origin with synthetic and repurposed compounds. Across studies, these selected compounds promote osteoblast commitment, differentiation, and matrix mineralization via BMP/Smad signaling and Wnt/beta-catenin (β-catenin) activation, often through glycogen synthase kinase-3 beta (GSK-3β) inhibition or relief of pathway antagonism or Hedgehog (Hh) pathway stimulation. Beyond osteoinduction, several candidates address issues that commonly limit repair, including angiogenesis, oxidative stress, inflammatory tone, osteoimmune regulation, and suppression of osteoclast-mediated resorption. Direct head-to-head comparisons are rare across both classes and reporting heterogeneity complicates interpretation. Key translational gaps include limited cytotoxicity and immunologic profiling, dose and release optimization, durability of benefit, and insufficient evaluation of rational combinations. More rigorous in vivo studies, including larger animal models and standardized outcome metrics, are needed to prioritize promising candidates and guide clinical development.
Background/Objectives: Ferroptosis, an iron-dependent form of regulated cell death defined by lipid peroxidation, has been extensively studied in cancer and neurodegeneration, but its contribution to erythropoiesis remains poorly understood. Methods: In this study, we investigated the expression of ferroptosis-related genes during HMBA-induced differentiation of murine erythroleukemia (MEL) cells and further assessed the effects of the ferroptosis inducer erastin in this model system. Results: HMBA treatment was accompanied by upregulation of ferroptosis-inducing genes (Atf3, Por, Tfrc, Slc11a2) and downregulation of inhibitory genes (Dhfr, Aifm2, Flvcr1, Nfe2l2, Slc3a2, Slc7a11), while Gpx4 levels increased. Erastin exposure identified 5 mu M as the optimal concentration, which resulted in a significant reduction of Steap3 transcripts, an increase in Hbb expression, and an increased accumulation of differentiated cells in culture, along with mild cytotoxicity. To be noted that at the protein level, erastin induced a similar to 10% decrease in STEAP3 and a 1.5-fold increase in beta-globin homo- or hetero-dimers. Ferroptosis markers confirmed erastin activity, with Fsp1 to be downregulated and Slc7a11, ferroportin, and the transferrin receptor upregulated. Importantly, erastin also enhanced apoptotic responses, as indicated by increased levels of active caspase-3 (similar to 40%) and reduced cellular proliferation rate (Ki-67, similar to 35%), suggesting overlap between ferroptotic and apoptotic pathways. Conclusions: Collectively, these findings indicate that erastin modulates erythroid maturation by repressing Steap3 (Six-transmembrane epithelial antigen of prostate 3) and enhancing Hbb expression, yet its differentiation inducing potential is counterbalanced by concurrent apoptosis activation. Overall, our results support a role of ferroptosis in erythroid maturation by linking iron metabolism, regulated cell death, and erythropoiesis, a fact of pharmacological and therapeutic relevance too.
Background/Objectives: Osteoporosis remains a clinically important metabolic bone disorder with limited bone-forming therapeutic options. SET domain bifurcated protein 1 (SETDB1) is involved in osteogenic epigenetic regulation, but small-molecule discovery guided by SETDB1-associated structural regions remains limited. This study aimed to identify a candidate compound with in silico relevance to a SETDB1-associated ligand-bound pocket and assess its association with early osteogenic readouts. Methods: A computational–experimental workflow was used, including hierarchical molecular docking, MM-GBSA rescoring, ADMET-based prioritization, redocking validation, molecular dynamics simulations, and preliminary in vitro evaluation in MC3T3-E1 cells. Compound 271 (C271) was selected based on structure-based screening results and predicted developability-related properties. Cytocompatibility, alkaline phosphatase (ALP) activity and staining, selected molecular markers, and SETDB1–H3 molecular dynamics behavior were evaluated. Results: Redocking reproduced the reference binding mode, and molecular dynamics simulations indicated that C271 maintained a relatively persistent conformation around the predicted SETDB1-associated pocket. Comparative SETDB1–H3 simulations showed altered H3 dynamics and SETDB1–H3 contact patterns in the C271-containing system. In cell-based assays, C271 showed no appreciable cytotoxicity within the tested concentration range and was associated with increased ALP activity and staining. C271 treatment was accompanied by higher global H3K9me3 and Runx2 levels, whereas SETDB1 protein abundance remained largely unchanged. Conclusions: C271 was identified as a computationally prioritized SETDB1-related candidate compound associated with early osteogenic-associated cellular responses. The evidence supports computational plausibility and cell-level association, but does not establish direct SETDB1 engagement, SETDB1 enzymatic modulation, SETDB1-dependent causality, or late-stage osteogenic maturation/mineralization. Given the single-compound evaluation, further target-engagement, enzymatic, and functional studies are needed.
Background: Ketoprofen is one of the most commonly prescribed NSAIDs; however, its pharmacogenetics remains poorly understood. The objective was to evaluate the influence of patients’ pharmacogenetic profiles on the effectiveness of ketoprofen for postoperative pain management after total hip arthroplasty, including postoperative analgesia (pain levels, opioid consumption) and the incidence of adverse reactions during hospitalization and up to 12 months post-surgery. Methods: The study included 53 patients (31 (58.49%) women, median age 66.0 [60.0–74.0] years) undergoing total hip arthroplasty. Genotyping was performed using real-time PCR to analyze 18 single-nucleotide polymorphisms (SNPs) across the following genes: CYP2C9 (rs1799853, rs1057910), CYP2C8 (rs10509681, rs11572080), CYP3A4 (rs35599367), CYP3A5 (rs776746), UGT2B7 (rs73823859, rs7439366, rs7668282), ABCB1 (rs1045642, rs4148738, rs2032582, rs1128503), PTGS1 (rs10306135, rs12353214), PTGS2 (rs20417), C3orf20 (rs12496846), and ZNF493-ZNF429 (rs2562456). Results: We did not find significant associations between patients’ genotypes and pain levels or postoperative opioid analgesic consumption or adverse reactions when ketoprofen was used for pain management in patients undergoing total hip arthroplasty. Conclusions: Routine pharmacogenetic testing for ketoprofen is not supported by our findings.
