Pharmaceuticals doi: 10.3390/ph19091403

Authors: Isidora Lazić Aljoša Stanković Nemanja Todorović Sofija Forkapić Mladena Lalić-Popović

Targeted Alpha Therapy (TAT) is often defined by the favorable physical properties of alpha particles, particularly their high linear energy transfer and short tissue range. However, these properties alone do not determine therapeutic success. The clinical behavior of alpha-emitting radiopharmaceuticals depends on whether radioactive decay can be matched to an appropriate biological scale and maintained within a chemically and pharmacokinetically coherent system. This review presents TAT as a multiscale design problem in which radionuclide production, radioactive decay, recoil, coordination chemistry, vector compatibility, tissue geometry, microdosimetry, biodistribution, and clinical outcome are interdependent rather than separate considerations. Established and emerging alpha emitters, including radium-223, astatine-211, lead-212/bismuth-212, actinium-225/bismuth-213, thorium-227 and terbium-149, are examined as distinct design solutions rather than interchangeable therapeutic options. Their comparison shows that no single radionuclide is universally optimal, as each occupies a distinct position within a landscape defined by physical, chemical and biological constraints. Viewing TAT as a multiscale design problem shifts radionuclide selection from the search for a universally superior emitter toward the rational matching of radionuclide properties to therapeutic context.

Pharmaceuticals doi: 10.3390/ph19091402

Authors: Chongyan Zhao Yuan Gao Ting Zhou Lei Sun Qingsong Qu

Metabolic diseases are jointly driven by insulin resistance, chronic inflammation, lipotoxicity, and disrupted organelle homeostasis arising from sustained nutrient overload. These disorders substantially increase the risk of cardiovascular, renal, and other multi-organ complications and have become a major global public health burden. Mitochondria are central organelles that integrate energy metabolism with stress signaling. They participate in fatty acid β-oxidation, the tricarboxylic acid cycle, and oxidative phosphorylation, while also regulating reactive oxygen species generation, mitochondrial DNA-related inflammatory signaling, calcium homeostasis, and cell death. Under chronic metabolic stress, the mitochondrial quality control (MQC) system shifts from adaptive repair toward decompensation, characterized by impaired mitochondrial biogenesis, abnormal mitochondrial dynamics, defective mitophagy, disrupted proteostasis and mitochondrial unfolded protein response, increased oxidative stress, and impaired metabolic reprogramming. Natural small molecules possess structural diversity and multi-target, multi-pathway regulatory properties. They can modulate multiple MQC processes and improve mitochondrial function and metabolic phenotypes in preclinical models. Novel formulations, structural optimization, and mitochondria-targeted delivery can further improve their solubility, bioavailability, tissue exposure, and subcellular localization, thereby enhancing therapeutic efficacy and translational potential. This review systematically summarizes the mechanisms of MQC dysregulation in metabolic diseases, the evidence supporting natural small-molecule interventions, and strategies for formulation and delivery optimization. Future studies should strengthen causal validation of MQC, quantify intramitochondrial drug exposure, and incorporate clinically relevant endpoints to facilitate the translation of natural small-molecule MQC modulators.

Pharmaceuticals doi: 10.3390/ph19091401

Authors: Dmitriy V. Ivashchenko Mikhail D. Che Farid R. Aysin Svetlana N. Tuchkova Ivan N. Korsakov Ekaterina I. Ianavichiute Mariia A. Ivashchenko Pavel V. Shimanov Rimma V. Kondratieva Artem V. Shubin Karin B. Mirzaev Yuriy S. Shevchenko Dmitry A. Sychev

Objectives. To identify associations between CYP3A4*22 and CYP3A5*3 genotypes, CYP2D6 phenotype, and the effectiveness and safety of antipsychotics in neurotypically developed boys with conduct disorders. Methods: The study included neurotypically developed boys aged 7–12 years who were hospitalized for conduct disorders. All patients were prescribed an antipsychotic. Patient follow-up lasted 14 days. Treatment effectiveness was assessed using a clinical aggression assessment (checklist) and the CGI-S, CGI-I, and CGAS scales. Safety was assessed using the UKU SERS and SAS scales. Patients were examined upon enrollment in the study, on day 5, and on day 14. All patients were genotyped for the CYP3A4*22 (rs35599367, C>T), CYP3A5*3 (rs776746, 6986T>C) CYP2D6*3 (rs35742686), CYP2D6*4 (G1846A, rs3892097), CYP2D6*6 (rs5030655), CYP2D6*10 (C100T, rs1065852), CYP2D6*41 (rs28371725) loci. CYP2D6 metabolism type was determined based on genotyping results, and patients were divided into two subgroups: those with normal metabolism (NM) and those with intermediate or poor metabolism (IM + PM). Results: Patients taking carbamazepine (n = 11) were excluded from the analysis of associations between treatment outcomes and the CYP3A4*22 and CYP3A5*3 polymorphisms. The analysis of associations between treatment outcomes and CYP2D6 metabolism type was conducted in two stages: the overall sample and a subsample of patients who were prescribed risperidone (n = 80). No significant associations were found between carrier status of the CYP3A4*22 and CYP3A5*3 polymorphisms and treatment effectiveness parameters. Analysis of the overall sample did not reveal any significant associations between CYP2D6 metabolism subtypes and the effectiveness parameters of drug therapy. Analysis of patients receiving risperidone revealed one statistically significant association: patients with CYP2D6 IM + PM reported headaches more frequently (16.1% vs. 2%; p = 0.03). Carrier status of the CYP3A4*22 polymorphism was significantly associated with asthenia and lethargy on day 5 (50% vs. 9.4%; p = 0.008). Conclusions: Our study identified only a few significant associations between the CYP3A4*22 polymorphism, CYP2D6 slow metabolism, and patients’ reports of early adverse reactions in a 14-day observation period. Further research is needed to identify pharmacogenetic predictors of the efficacy and safety of antipsychotics in neurotypical children with conduct disorders.

Pharmaceuticals doi: 10.3390/ph19091400

Authors: Mohammed A. Akeel Ekramy M. Elmorsy Fahad M. Alshammari Aly A. M. Shaalan Abdulrahman S. Aldaghmi Barakat M. Alrashdi Saad M. Alrashidi Gehad E. Elshopakey Baraah Abu Alsel Manal S. Fawzy

Background/Objectives: Oxidative stress, chronic inflammation, disrupted apoptosis, and altered autophagy are biological processes implicated in colorectal tumor development. Morin is a plant-derived flavonoid with antioxidant and anti-inflammatory properties, but its limited solubility and bioavailability may limit its biological activity. This study evaluated the effects of free morin and morin-loaded liposomes (MOR-Lips) on 1,2-dimethylhydrazine (DMH)-associated colonic biochemical, molecular, and histopathological alterations in rats. Methods: Rats were randomly assigned to six groups—vehicle control, MOR, MOR-Lips, DMH, DMH + MOR, and DMH + MOR-Lips—and treated for 10 weeks. Serum and colonic tissues were evaluated for cancer-associated biomarkers (CEA, CA19-9, CA125, HMG-CoA reductase), oxidative stress and antioxidant indices, nitrosative and oxidative DNA-damage markers (MDA, NO, 8-OHdG), inflammatory mediators (TLR4/NF-κB/COX-2, cytokines, MPO), proliferative indices (Ki-67, PCNA), apoptotic and autophagy-related regulators (Bax, caspase-3, p53, cytochrome c, BCL-2, p-AKT, LC3-II, Beclin-1, p62), and histopathological and immunohistochemical changes. Results: DMH exposure was associated with increased CEA, CA19-9, CA125, HMG-CoA reductase, MDA, NO, 8-OHdG, TLR4/NF-κB/COX-2, cytokines, MPO, Ki-67, and PCNA. DMH also reduced NRF2/HO-1 signaling and antioxidant defenses, shifted apoptosis-related markers toward a pro-survival profile, altered autophagy-related markers, and produced marked colonic histopathological abnormalities. Both free MOR and MOR-Lips attenuated several of these DMH-associated alterations, with MOR-Lips generally producing greater effects than free MOR. MOR-LIP treatment was associated with restoration of antioxidant marker profiles, reduced levels of inflammatory and proliferative markers, a shift toward a pro-apoptotic marker profile, partial normalization of autophagy-related markers, and improved colonic histopathological appearance. Conclusions: In DMH-exposed rats, MOR-Lips were associated with more favorable redox, inflammatory, proliferative, apoptosis-related, autophagy-related, and histopathological profiles than free MOR. These findings support further investigation of MOR-Lips as a formulation strategy for improving the biological activity of morin. Because quantitative preneoplastic and neoplastic endpoints were not measured, the results do not establish inhibition of colorectal carcinogenesis or chemopreventive efficacy.

Pharmaceuticals doi: 10.3390/ph19091399

Authors: Greg Haroutunian Lawrence Daniels Ashot Tsaghikian Caifeng Zhao Phaedon Zavras Svetlana Marukian Haiyan Zheng Arevik Mosoian

Background: Triple-negative breast cancer (TNBC) remains a major cause of cancer mortality due to its aggressive behavior, metabolic adaptability, and high therapeutic resistance. Extracellular vesicles (EVs) within the tumor microenvironment contribute to tumor progression and metastasis by transferring pro-tumorigenic cargo. While conventional Tumor Treating Fields use high-frequency alternating fields to disrupt mitosis, low-energy triple-frequency bioelectromagnetic approaches remain poorly characterized. Methods: We evaluated a device–drug strategy combining triple-frequency low-intensity electromagnetic stimulation (EMS2: 396 Hz, 285 Hz, 528 Hz) with the pleiotropic drug Fingolimod (FTY720). Treatments were tested in MDA-MB-231 and ARM-G breast cancer cells, with Paclitaxel as a positive control. Cell proliferation was assessed by MTS assay, and extracellular vesicles were isolated following individual and combination treatments. Quantitative LC-MS/MS proteomics was used to characterize treatment-induced changes in EVs cargo. Results: EMS2 reduced proliferation in both cell lines and produced morphological changes consistent with altered cell-cycle progression. EMS2 alone triggered adaptive metabolic responses, whereas combination with Fingolimod suppressed these compensatory signatures. EVs proteomics revealed combination-specific alterations associated with mitochondrial stress, ER stress, NF-κB suppression, and autophagy-associated pathways. The combination also reduced levels of metastasis- and stroma-associated proteins, including Mitogen-Activated Protein Kinase 12 (MAPK12) and collagen-associated ECM components (Collagen Type I Alpha 1 Chain (COL1A1), Collagen Type VI Alpha 1 Chain (COL6A1), Collagen Type VI Alpha 3 Chain (COL6A3), and Matrilin 3 (MATN3)) in EVs. Bliss independence analysis identified a subset of metastasis-associated proteins suppressed in EVs beyond the level predicted by an additive model, an exploratory finding that will require further validation with dose–response and functional assays. Conclusions: Combined triple-frequency EMS2 and Fingolimod treatment altered the extracellular vesicle proteome, inducing signatures consistent with mitochondrial and endoplasmic reticulum stress, metabolic disruption, and reduced levels of metastasis-associated and stromal/ECM remodeling proteins, along with reduced proliferation. These findings suggest a coordinated anti-cancer effect of this tunable device–drug strategy, warranting further functional and in vivo validation to confirm therapeutic potential.

Pharmaceuticals doi: 10.3390/ph19091398

Authors: Tianle Zhan Haoran Chen Jian Yao Chuangqi Yu

Background/Objectives: Sjögren’s Syndrome (SS) is an autoimmune disorder with impaired exocrine gland function. Its pathogenesis remains elusive, but aberrant innate/adaptive immunity and dysregulated interferon signaling are core features. Current SS research lacks exploration of innate immune mechanisms and therapeutic targets. Phloretin, a natural dihydrochalcone derivative, inhibits excessive innate immune activation, yet its role and mechanism in SS remain unreported. Methods: This study aimed to explore the therapeutic efficacy of phloretin against Sjögren’s syndrome (SS) and elucidate the regulatory mechanism underlying its effects on dendritic cells (DCs) and the Stimulator of Interferon Genes (STING)-driven innate immune cascade. Clinical specimens were utilized to analyze DC infiltration and STING expression in the labial glands of SS patients. In vivo experiments were conducted on NOD/Ltj SS mice with phloretin treatment, using hydroxychloroquine as a positive control. For in vitro studies, DCs were stimulated with poly I:C to verify the functional effects of phloretin and its regulatory role in the STING/TANK-binding kinase 1 (TBK1)/Interferon Regulatory Factor 3 (IRF3) signaling. Salivary secretion, glandular inflammatory responses, and systemic immune overactivation were evaluated in SS mice. In poly I:C-stimulated DCs, cell activation, migration, and co-stimulatory molecule expression were measured. The regulatory effects of phloretin on the STING signaling cascade and STING ubiquitin-mediated degradation were examined by Western blotting. Phloretin, initially identified as a candidate compound via virtual screening, was subjected to thermal shift assay (CETSA), drug affinity responsive target stability (DARTS) and molecular dynamics simulation (MD) for direct binding verification with STING. The STING overexpression rescue experiment further corroborates that phloretin modulates innate immune responses in a STING-dependent manner. Results: Clinical results revealed obvious infiltration of CD11c+STING+ DCs and CD11c+p-TBK1+ DCs in labial gland lesions of SS patients. In NOD/Ltj SS mice, phloretin treatment recovered salivary secretion, relieved glandular inflammatory injury, and restrained excessive activation of innate and adaptive immune responses, showing comparable therapeutic effects compared with hydroxychloroquine. In vitro experiments confirmed that phloretin could alleviate poly I:C-induced abnormal activation and migration of DCs, as well as reduce the expression of co-stimulatory molecules. Mechanistically, phloretin was found to interfere with the STING signaling cascade, alter the ubiquitination level of STING protein, and further inhibit the abnormal activation of DCs, which may contribute to its protective effects against Sjögren’s syndrome. Notably, results from MD, CETSA and DARTS collectively validate the interaction between phloretin and STING protein, providing solid molecular-level evidence for its regulatory effect on STING signaling transduction. Conclusions: DCs and STING-driven innate immunity critically contribute to SS progression. Phloretin effectively ameliorates SS-related immune disorders by restraining DC overactivation and STING signaling, supporting it as a promising candidate agent for SS treatment.

Pharmaceuticals doi: 10.3390/ph19091397

Authors: Shunbo Zhao Bingda Wu Hui Shen Qi Zhou Chang Shu Li Ding

Background/Objectives: AC02 is a novel 39-amino-acid adrenocorticotropic hormone (ACTH) analogue designed for the treatment of infantile spasms. To support its clinical study, in which porcine ACTH1–39 served as the positive-control drug, reliable methods for the determination of AC02 and porcine ACTH1–39 in human plasma were required. Reported analytical methods for ACTH analogues are mainly immunoassays, which are easily affected by cross-reaction and the hook effect, necessitating a more selective analytical approach. Methods: Two LC-MS/MS methods were developed for the determination of ACTH analogues AC02 and porcine ACTH1–39 in human plasma. Human ACTH1–39 was included as a selectivity marker to confirm that endogenous ACTH does not interfere with the quantification of AC02. Based on the distinct concentration requirements and matrix challenges, two sample-preparation procedures were established: micro-solid-phase extraction coupled with protein precipitation for porcine ACTH1–39 (LLOQ 0.100 ng/mL), and acid-mediated protein precipitation for AC02 (LLOQ 0.500 ng/mL). The [M+6H]6+ ions were selected as precursor ions, and the corresponding 5+ fragment ions, formed by loss of the C-terminal phenylalanine, were used for MRM detection. Results: Despite a mass difference of only 0.98 Da between AC02 and human ACTH1–39, which are indistinguishable by mass spectrometry, baseline chromatographic separation was achieved. Both methods were fully validated in accordance with current bioanalytical guidelines. Conclusions: The validated methods were successfully applied to the phase I clinical study of AC02 and porcine ACTH1–39, enabling reliable quantification of the drug candidate and its active comparator, porcine ACTH1–39.

Pharmaceuticals doi: 10.3390/ph19091396

Authors: Mostafa M. El-Sheekh Eman Bases Shimaa M. El Shafay Rania A. El-Shenody Mostafa E. Elshobary Abdel Hady A. Abdel Wahab Wesam E. Yousuf Dorya I. Essa Samar Sami Alkafaas

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Pharmaceuticals doi: 10.3390/ph19091395

Authors: Luciane Mendes Monteiro Manuel Minteguiaga Vera Lucia Pereira dos Santos Sara Emilia Lima Tolouei Shabana I. Khan Mei Wang Joseph Lee Wilmer H. Perera Paulo Vitor Farago Farhan Mahmood Shah Abbas Ali Daniele Monteiro Ikhlas Ahmed Khan Sílvia Beatriz Bürger Tinelli Arquimedes Gasparotto Junior Jane Manfron

Background: Brazilian Piper species are chemically diverse sources of volatile metabolites, yet direct comparative datasets integrating complementary analytical and biological endpoints remain limited. Methods: This study compared hydrodistilled essential oils (EOs) from Piper amalago, P. diospyrifolium, P. grazielae, and P. solmsianum using GC-MS, HPTLC fingerprinting, qualitative HPTLC-DPPH visualization, in vitro cytotoxicity screening, and behavioral assays with imported fire ants. Results: GC-MS revealed distinct chemical profiles among the four analyzed EOs. P. solmsianum was dominated by monoterpenes, mainly β-pinene (28.21%) and myrcene (24.73%); P. diospyrifolium was characterized by trans-β-caryophyllene (22.06%); and P. grazielae by germacrene D (14.06%), trans-β-caryophyllene (12.53%), and bicyclogermacrene (9.50%). P. amalago showed a mixed terpene profile containing bicyclogermacrene (11.00%), spathulenol (9.21%), and β-phellandrene (8.12%). HPTLC generated visually discriminative fingerprints that complemented the GC-MS profiles and identified chromatographic zones for subsequent compound-level characterization. Qualitative HPTLC-DPPH visualization revealed DPPH-reactive zones, most prominently in P. grazielae. The EOs produced moderate, cell-line-dependent cytotoxic responses, with P. grazielae showing the lowest IC50 values against BT-549 (45 µg/mL) and SK-OV-3 (43 µg/mL) cells and modest apparent selectivity indices (1.56–1.72). P. amalago reduced sand removal by hybrid imported fire ants at 31.25 µg/g, whereas P. solmsianum reduced sand removal by hybrid and red imported fire ants at 62.5 and 31.25 µg/g, respectively; neither oil caused mortality at 125 µg/g. Conclusions: The four EOs showed distinct chemical and biological profiles under the standardized conditions employed. P. grazielae showed the lowest IC50 values in the cytotoxicity screening and the most prominent DPPH-reactive chromatographic response, whereas P. amalago and P. solmsianum produced measurable nonlethal deterrence of fire ant digging behavior.

Pharmaceuticals doi: 10.3390/ph19091394

Authors: Moaz A Eltabeeb Rofida Albash Mariam Hassan Sadek Ahmed Ahmed M. Agiba Azza A. K. El-Sheikh Diana E. Aziz

Background/Objectives: Cutaneous fungal infections remain a major therapeutic challenge due to poor skin penetration and emerging antifungal resistance of current treatments. Drug repurposing offers a promising strategy to expand antifungal therapy, with ketoprofen (KPN) recently demonstrating antifungal activity. Methods: In this study, KPN-cationic aspasomes (Ca-ASPMs) were developed using a Quality by Design (QbD) approach to enhance topical delivery and maximize the therapeutic potential of repurposed KPN. A D-optimal experimental design was implemented to investigate the influence of ascorbyl palmitate amount (X1), ethanol concentration (X2), and cationic SAA type (X3) on the critical quality attributes of KPN-Ca-ASPMs, namely entrapment efficiency, particle size, and zeta potential. Results: Numerical optimization identified an optimum formulation comprising 10 mg ascorbyl palmitate, 5% ethanol, and didodecyldimethylammonium bromide (DDAB) as cationic SAA, with an overall desirability of 0.815. The optimal KPN-Ca-ASPM (F9) exhibited nanosized vesicles (213.24 ± 2.20 nm) and 91.30 ± 10.01% entrapment efficiency. TEM confirmed the spherical morphology of the vesicles, while DSC demonstrated successful incorporation of KPN within the aspasomal matrix. F9 exhibited enhanced in vitro drug release (68% after 6 h) and excellent storage stability. Confocal laser scanning microscopy demonstrated enhanced skin penetration of F9. In a murine cutaneous candidiasis model, F9 significantly enhanced the antifungal efficacy of KPN, producing a 2.989-log reduction in fungal burden compared with the untreated group and significantly outperforming the free drug (p = 0.0005). Histopathological examination confirmed restoration of normal skin architecture. Conclusions: Collectively, the QbD-guided development of KPN-Ca-ASPM provided a reproducible nanocarrier that significantly enhanced the topical antifungal efficacy of repurposed KPN, highlighting its potential for the treatment of cutaneous candidiasis.

Pharmaceuticals doi: 10.3390/ph19091393

Authors: Shuo Yan Hui Yang Yingrui Wang Lejian Zhu Binsheng Wang Leiming Zhang Qing Hao

Metabolic dysfunction-associated steatotic liver disease (MASLD) develops when hepatic lipid acquisition and synthesis exceed the capacity for oxidation and very-low-density lipoprotein export. Flavonoids act on several components of this network, yet their therapeutic development is constrained by poor aqueous solubility, extensive intestinal and first-pass metabolism, variable activity of circulating metabolites, and limited information on hepatic exposure. Experimental studies link representative flavonoids to AMPK–SREBP-1c and PPARα signaling, mitochondrial quality control, Nrf2-dependent redox defense, inflammatory pathways, and the gut–liver axis. By contrast, the available randomized trials of quercetin, hesperidin, anthocyanins, green-tea catechins, EGCG, and soy isoflavones show at most modest changes in liver fat or biochemical markers and do not demonstrate metabolic dysfunction-associated steatohepatitis (MASH) resolution or fibrosis regression. Liposomal, lipid-based, polymeric, and nanocrystal formulations have improved dissolution, systemic exposure, or liver distribution in preclinical models, but comparative pharmacokinetics, chronic safety, manufacturability, and clinical efficacy remain poorly defined. The evidence therefore supports viewing flavonoids as formulation-dependent investigational candidates rather than established MASLD therapies. Progress will depend on chemically standardized products, exposure–response studies, clinically relevant models, and adequately powered trials using validated imaging or histological endpoints.

Pharmaceuticals doi: 10.3390/ph19091392

Authors: İbrahim Gecili Muhammed Sait Ertuğrul Irmak Ferah Okkay Onur Şenol Ufuk Okkay Mustafa Özkaraca Ziadoon Al-Yaqoobi Cemil Bayram A. M. Abd El-Aty Ali Taghizadehghalehjoughi Ahmet Hacımüftüoğlu

Background: Alterations in glutamatergic homeostasis have been implicated in Alzheimer’s disease, but the relationship between total hippocampal tissue glutamate and Alzheimer-like molecular alterations remains unclear. This study evaluated the early neuroprotective potential of gentisic acid (GA) and erucic acid (EA) in an intracerebroventricular streptozotocin (icv-STZ)-induced rat model of sporadic Alzheimer-like pathology. Methods: Sixty-four female Sprague–Dawley rats were randomly assigned to eight groups: control, sham, STZ, STZ + GA (100 or 200 mg/kg/day), STZ + EA (25 or 50 mg/kg/day), and STZ + memantine (10 mg/kg/day). Treatments began immediately after STZ and continued orally for 21 days. Behavioral performance was assessed by Morris water maze and passive avoidance tests. Total hippocampal tissue glutamate was quantified by LC–MS/MS. Neuronal degeneration was assessed histopathologically, whereas tau, amyloid-β, and AChE immunoreactivity and tau-, APP-, and AChE-related in situ hybridization signals were evaluated. Results: Compared with controls, STZ-treated rats exhibited poorer behavioral performance, higher total hippocampal tissue glutamate, greater neuronal degeneration, and increased molecular signals. GA and EA administration was associated with attenuation of these alterations, with generally stronger effects at higher doses. EA at 50 mg/kg/day showed the lowest mean total hippocampal tissue glutamate among treatment groups. GA at 200 mg/kg/day and EA at 50 mg/kg/day showed the most consistent preservation across behavioral, histopathological, and molecular endpoints. Conclusions: Early GA and EA administration was associated with neuroprotective effects in the icv-STZ model. Because treatment began immediately after STZ administration, the findings support early or preventive neuroprotection rather than reversal of established Alzheimer-like pathology. Total tissue glutamate measurements neither distinguish extracellular glutamate nor directly demonstrate glutamate-mediated excitotoxicity.

Pharmaceuticals doi: 10.3390/ph19091391

Authors: Emma Julin Pongoh Rymond Jusuf Rumampuk

Background/Objectives: In the search for potent non-sugar α-glucosidase inhibitors with improved safety profiles, a novel flavonoid glycoside was isolated for the first time from the stem bark of Albizia saponaria (Fabaceae). The objective of this study was to elucidate its chemical structure and evaluate its therapeutic potential as an anti-hyperglycemic agent compared to known related flavonoids and a standard clinical drug. Methods: Comprehensive structural elucidation was performed using high-resolution mass spectrometry and multidimensional 1D/2D NMR (1H, 13C, HSQC-DEPT, COSY, and CIGAR). To assess its inhibitory efficacy and pharmacokinetic profiles, an in silico comparative study was conducted against a database of related flavonoids (Quercitrin, Hyperoside, and Isoquercitrin) and the clinical drug Acarbose. This involved molecular docking simulations against human intestinal maltase-glucoamylase (PDB ID: 3TOP) alongside integrated ADMET modeling and toxicological screening. Results: The compound was successfully identified as 4′,7-dihydroxyflavan-3′-O-β-D-glucoside (1). Molecular docking revealed that Compound 1 exhibited a superior predicted binding affinity of −9.5 kcal/mol, outperforming Quercitrin (−9.3 kcal/mol), Hyperoside (−8.3 kcal/mol), Isoquercitrin (−7.9 kcal/mol), and Acarbose (−7.2 kcal/mol). This strong thermodynamic stability is driven by a robust conventional hydrogen-bonding network with key active site residues (Arg1377, Gln1372, and Gly1365), successfully overriding a localized electrostatic strain at Asp1279. Furthermore, ADMET modeling demonstrated a highly desirable local pharmacokinetic framework; its low Caco-2 permeability (−6.432) and low human intestinal absorption (HIA = 0.120) favor targeted luminal retention in the gastrointestinal tract, mirroring Acarbose while minimizing systemic exposure. Crucially, toxicological screening unveiled a significant safety advantage for Compound 1, marked by negligible CYP3A4 interaction (0.004) and a remarkably low risk of Drug-Induced Liver Injury (DILI = 0.213) compared to the high-risk hepatotoxic profile of Acarbose (DILI = 0.882) and the reference flavonoids (DILI > 0.69). Conclusions: These predictive findings establish Compound 1 as a highly promising, low-toxicity natural scaffold for anti-hyperglycemic drug development. Its superior binding affinity and minimized hepatotoxicity risk warrant subsequent in vitro and in vivo functional validation.

Pharmaceuticals doi: 10.3390/ph19091390

Authors: Debdulal Chakraborty Tushar Kanti Sinha Rupak Kanti Biswas Aniruddha Maiti Manab Jyoti Barman Ranabir Bhattacharya Subhendu Kumar Boral Arnab Das

Purpose: To evaluate the incidence, predictors, and clinical impact of treatment-emergent epiretinal membrane (ERM) in diabetic macular edema (DME) treated with dexamethasone implant, aflibercept, ranibizumab, bevacizumab, or faricimab. Methods: In this multicenter, retrospective, nonrandomized study, 501 eyes with center-involving DME were assessed and 423 eyes without ERM on pretreatment OCT and with 24-month follow-up were included. Eyes were classified by the intravitreal agent selected by the treating physician in routine care and maintained throughout follow-up; eyes requiring a drug switch or retinal laser/vitrectomy during follow-up were excluded. The last eligible pre-index SD-OCT confirmed the absence of ERM; BCVA, central subfield thickness (CST), and vitreomacular-interface (VMI) morphology were assessed at baseline and 6, 12, 18, and 24 months. Crude comparisons, multivariable logistic regression, and Kaplan–Meier/log-rank analyses were performed. Results: ERM developed in 36/423 eyes (8.5%). Incidence was 16.0% with dexamethasone implant, 7.5% with aflibercept, 7.7% with ranibizumab, 7.4% with bevacizumab, and 4.3% with faricimab. Dexamethasone implant showed higher ERM odds than pooled non-dexamethasone therapy after adjustment (OR 2.24, 95% CI 1.02–4.92; p = 0.044). Baseline VMI abnormality, severe NPDR/PDR, prior PRP, and higher baseline CST were also associated with ERM. Eyes developing ERM had smaller BCVA gains (+2.8 vs. +7.1 ETDRS letters; p < 0.001) and CST reductions (88.6 vs. 130.8 µm; p = 0.004). Conclusions: Treatment-emergent ERM was associated with baseline ocular characteristics and treatment category and with poorer 24-month outcomes. These observational drug-group signals require prospective confirmation.

Pharmaceuticals doi: 10.3390/ph19091389

Authors: Yixiang Pan Yiting Zhu Wenjie Chen Jie Xu Jinhang Ma Odinaka Cassandra Ezekiel Hailv Ye Jiang Wu

Background/Objectives: Conventional surgical sutures can promote local coagulation after implantation, potentially compromising vascular patency. Heparin is an effective anticoagulant, but systemic administration may cause bleeding and thrombocytopenia. This study aimed to develop an electrospun heparin-loaded polycaprolactone (PCL) suture that combines structural support with sustained local heparin delivery and antithrombotic activity. Methods: Heparin was incorporated into electrospun PCL sutures at nominal heparin-to-PCL feed ratios of 0.05 and 0.50 wt%. Fiber morphology, chemical characteristics, tensile properties, heparin release, molecular interactions, cytocompatibility, hemolysis, and plasma recalcification were evaluated. Molecular dynamics simulations were used to investigate PCL–heparin interactions. In vivo antithrombotic performance was assessed in a liver puncture model using male C57BL/6 mice, followed by histological examination. Results: Heparin incorporation preserved the fibrous morphology of the sutures, although maximum tensile stress and strain were modestly reduced compared with PCL alone. Both heparin-loaded formulations exhibited sustained heparin release over 10 days, and molecular dynamics simulations indicated stable PCL–heparin interactions. Cell viability did not differ significantly among the suture groups, and all hemolysis rates remained below 5%. In the plasma recalcification assay, both heparin-loaded groups remained incompletely coagulated at 300 s, indicating marked anticoagulant activity. In the mouse liver injury model, heparin-loaded sutures reduced local clot formation compared with PCL sutures. Conclusions: Electrospun heparin-loaded PCL sutures integrate sustained local heparin delivery with in vitro anticoagulant and in vivo antithrombotic activity. These findings support further evaluation of this multifunctional suture platform in clinically relevant vascular models.

Pharmaceuticals doi: 10.3390/ph19091388

Authors: Linfeng Li Jiajia Li Li Yang Jun Zhou Yuying Ma

Chemically induced proximity (CIP) has revolutionized small-molecule pharmacology by moving beyond traditional occupancy-based inhibition and activation. Within this paradigm, monovalent nondegrading molecular glues (ndMGs) represent a structurally compact and mechanistically distinct class of linkerless compounds. These agents stabilize or induce selective biomolecular interactions, altering the functional state or cellular localization of macromolecular complexes without causing component degradation. Once considered rare phenomena unique to specific natural products, ndMGs are now being actively engineered across diverse therapeutic landscapes, with multiple candidates advancing into clinical trials and achieving regulatory validation. We systematically highlight their evolving capabilities to modulate oncogenic networks, intervene with immune activity, rectify metabolic signaling, control neurodegenerative trafficking and stress networks, and disrupt critical pathogen assemblies, while broadening the druggable landscape into non-canonical mechanisms. In each section, we analyze the therapeutic value of the targets, the molecular mechanisms of action, the latest progress, and the unique challenges faced by these strategies. In summary, the ndMG modality represents a highly versatile strategy to unlock the undruggable proteome.

Pharmaceuticals doi: 10.3390/ph19091387

Authors: Boryana Nikolova-Mladenova Rositsa Mihaylova Stilyana Kostova Boris Vasilev Irini Doytchinova Mariyana Atanasova

Background: Building on earlier work with methoxy-, dimethoxy-, bromo-, and nitro-substituted salicylaldehyde benzoylhydrazones, we synthesized and evaluated new 4-chloro- and 5-chloro derivatives bearing an additional chlorine substituent on the hydrazide-derived phenyl ring to further explore the impact of halogenation on cytotoxic activity. Methods: Chloro-hydrazones were obtained through a one-step condensation of 4- or 5-chlorosalicylaldehyde with benzhydrazide or 2-, 3-, and 4-chlorobenzhydrazides. Their physicochemical, pharmacokinetic, ADME, lead-likeness, and drug-likeness profiles were evaluated in silico using SwissADME, ACD/Labs v9.10, and MDL QSAR v2.2.0.0.446. The synthesized compounds were structurally characterized by IR, 1H NMR, 13C NMR, and HR ESI–MS, and their cytotoxic activity was subsequently assessed by the MTT assay in selected cancer cell lines. Molecular docking was performed against ABL1 tyrosine kinase (ABL1 TK), a potential molecular target relevant to two of the investigated leukemia cell lines, using GOLD v.5.2.2 (CCDC Ltd., Cambridge, UK). Results: The chloro-substituted salicylaldehyde benzoylhydrazones exhibited pronounced, cell-type-dependent cytotoxicity, with leukemia cells being substantially more sensitive than breast carcinoma cells. Most derivatives showed low- to sub-micromolar IC50 values against SKW-3 human T-cell prolymphocytic leukemia, K-562 human chronic myeloid (myelogenous) leukemia, and BV-173 human BCR::ABL1-positive leukemia cells, with K3 and K4 exhibiting the highest activity in SKW-3 cells (IC50 = 0.5 ± 0.1 µM). K5 showed particularly strong activity against K-562 and BV-173 cells (IC50 = 0.7 ± 0.1 and 0.9 ± 0.1 µM, respectively), compared with 26.9 ± 2.4 and 21.5 ± 3.3 µM for imatinib. HL-60 human acute promyelocytic leukemia cells showed intermediate sensitivity (IC50 = 1.3–13.9 µM), whereas the activity against MCF-7 estrogen receptor-positive and MDA-MB-231 triple-negative breast carcinoma cells was more variable (1.5–32.1 and 4.1–62.5 µM, respectively). The derivatives showed high selectivity toward malignant cells relative to non-malignant CCL-1 non-malignant mouse fibroblasts, with lower-bound SI values frequently exceeding 50 and reaching >200 in SKW-3 cells. Conclusions: The chloro-substituted salicylaldehyde benzoylhydrazones exhibited pronounced and selective cytotoxicity, particularly toward leukemia cell lines, identifying this scaffold as a promising starting point for further anticancer drug-discovery studies.

Pharmaceuticals doi: 10.3390/ph19091386

Authors: Yajun Zhou Bangjian Liu Hui Wang Li Cao

Introduction: Alzheimer’s disease (AD) is a chronic progressive neurodegenerative disorder involving ROS and ferroptosis. MXene, a 2D material, functions as both a drug carrier and ROS scavenger. Butylphthalide (NBP), a neuroprotective agent, suffers from poor aqueous solubility and bioavailability. This study synthesized Ti2C@BSA-NBP, a novel MXene nanocomposite, to enhance NBP delivery and therapeutic efficacy for AD. Materials and Methods: Ti2C@BSA-NBP was synthesized via self-assembly and amidation reaction, and characterized by multiple techniques. In vitro, ROS-scavenging capacity, mitochondrial function, and ferroptosis markers were assessed in H2O2-injured cells. In vivo efficacy was evaluated in an AD mouse model via behavioral, histopathological, and biochemical analyses. Results: The nanocomposite exhibited robust ROS-scavenging activity in vitro, significantly attenuating ROS, restoring mitochondrial function, and reversing ferroptosis markers. In vivo, Ti2C@BSA-NBP ameliorated learning/memory deficits, partially repaired neuronal morphology, and suppressed neuroinflammation. Mechanistically, it downregulated ACSL4 and Aβ overexpression while restoring GPX4 inhibition. Discussion: Ti2C@BSA-NBP exerts synergistic neuroprotection through MXene-mediated ROS clearance and NBP-mediated multi-target regulation, counteracting oxidative stress, ferroptosis, and neuroinflammation. Conclusions: Ti2C@BSA-NBP is a promising multifunctional nanoplatform integrating antioxidant activity, enhanced drug delivery, and ferroptosis modulation for AD therapy.

Pharmaceuticals doi: 10.3390/ph19091385

Authors: Junge Li Yuxin Liu Xin Shao Yuanlu Zhang Zhidong Qiu Yongchun Wang Feiran Qi Qiuzhu Tang Ailing Jia

Background: Primary dysmenorrhea (PD) is a prevalent gynecological condition that significantly compromises the quality of life in adolescents and women of reproductive age. Within traditional Chinese medicine (TCM), Cold Coagulation and Blood Stasis Primary Dysmenorrhea (CCBS-PD) represents the most frequently observed syndrome pattern of PD. Wenjing Decoction (WJD), a classical TCM formulation, has been extensively employed for the treatment of CCBS-PD. However, given the multi-component and complex nature of WJD, the potential mechanisms underpinning its therapeutic effect have yet to be elucidated. Methods: A rat model of CCBS-PD was induced through ice-water bath stimulation in conjunction with estradiol benzoate and oxytocin. The pharmacological effects of WJD were evaluated by writhing response, hemorheological parameters, uterine index, histopathological examination, and biochemical assays. Serum-exposed constituents of WJD were characterized using UPLC-Orbitrap Exploris 120 MS. To study the mechanisms of WJD, methods from network pharmacology, molecular docking, and off-target metabolomics were used. Western blot analysis examined representative proteins in the signaling pathways predicted by network pharmacology. Network pharmacology and metabolomics were integrated to construct a pathway–metabolite–target–compound network, and representative targets were analyzed by RT-qPCR. Results: WJD treatment reduced writhing responses, improved hemorheological abnormalities, and alleviated uterine pathological changes in CCBS-PD rats. Serum pharmacochemistry identified 62 WJD-derived constituents. Metabolomics analysis indicated that WJD was associated with alterations in nitrogen metabolism, valine/leucine/isoleucine biosynthesis and arginine biosynthesis. Network pharmacology and molecular docking suggested several candidate compounds and targets, including Robinetin, Levistolide A, Pratol, 7,4′-dihydroxyflavone, EGFR, AKT1, ESR1, MMP9, MAPK3, and TNF. RT-qPCR showed that selected genes, including AKT1, EGFR, MAPK3, TNF, ESR1, MMP9 and CASP3, were changed in the model group and partially restored following WJD improvement. Western blot analysis demonstrated that WJD treatment decreased the phosphorylation levels of AKT, ERK1/2, and NF-κB, and down-regulated the protein expression of COX-2. Conclusions: WJD showed beneficial effects in a CCBS-PD rat model. Synthesized assessments indicate a potential link to the actions of serum-exposed constituents, alterations in amino acid metabolism, and modulation of inflammation-related signaling pathways. This research offers initial indications suggesting the multi-component and multi-target pharmacological actions of WJD, although the underlying mechanisms remain to be validated through targeted metabolomics and functional studies.

Pharmaceuticals doi: 10.3390/ph19091384

Authors: Usman Ali Shams Fawad Inayat Muhammad Asif Zeb Maryam Sulaiman Shams Muhammad Jawad Ullah Silvia Jiménez-Morales

Acute myeloid leukemia (AML) represents a heterogeneous group of hematological malignancies characterized by uncontrolled proliferation of myeloid progenitors and accumulation of immature blasts in the bone marrow. Metabolic reprogramming is now recognized as a core hallmark of AML, generating dependencies that distinguish leukemic cells from normal hematopoietic stem and progenitor cells and that can be exploited therapeutically. In this review we follow a single connected line of argument: we first place metabolic rewiring within the broader hallmarks of cancer, then describe the principal metabolic programs altered in AML and the specific features that distinguish AML from other malignancies. We next examine the inhibitors and drugs that target each of these pathways, linking every drug class to its mechanism of synergy with chemotherapy, the preclinical and clinical evidence available, and its association with outcome in AML. We then consider multi-target (combination) therapy as a distinct opportunity, and finally the principal challenges that remainsafety and tolerability, the metabolic heterogeneity and plasticity of AML, and the design of biomarker-guided trials. Multiple classes of metabolic drugs are discussed, including glycolysis inhibitors, oxidative phosphorylation inhibitors, glutamine metabolism antagonists, fatty acid oxidation modulators, and redox-active compounds. Despite significant challenges, targeting cellular metabolism represents a promising strategy to enhance therapeutic outcomes in patients with AML.

Pharmaceuticals doi: 10.3390/ph19091383

Authors: Emanuela De Bellis Danilo Donnarumma Annarita Pagano Ines Mazza Bruno Charlier Simona Madonia Alfonso Masullo Mirella Onofrio Valentina Fortunato Anna Maria Spera Federica Campana Gianluigi Franci Flora Salzano Graziamaria Corbi Amelia Filippelli Pasquale Pagliano Valeria Conti

Background/Objectives: People Living with HIV (PLWH) often have comorbidities require polypharmacy, increasing the risk of adverse events (AEs) associated with drug–drug/drug–supplement interactions (DDIs/DSIs). However, DDIs and DSIs, including those involving antiretrovirals (ARVs), and their related effects have not been exhaustively assessed. This study aimed to determine, in HIV patients on HAART who are taking multiple medications, the prevalence of DDIs and DSIs involving both ARVs and non-ARV co-medications, and their clinical relevance and consequences. Methods: In this longitudinal observational study, PLWH receiving HAART were consecutively enrolled at the University Hospital of Salerno and followed up during scheduled visits. Demographic, clinical, and pharmacological data were collected. DDIs/DSIs were identified using Drugs.com, Lexicomp, and the Liverpool HIV Interaction Checker. Interactions classified as major (Drugs.com), consider therapy modification or avoid combination (Lexicomp), and do not coadminister (Liverpool) were deemed highly clinically relevant. This assessment was supplemented with review of SmPCs and the available literature. Results: Among 84 PLWH, 229 interactions were identified, and 37/229 (16.2%) were highly clinically relevant. Eight patients (9.5%) experienced AEs predicted by the tools. Four were DDIs: cobicistat/alprazolam associated with sedation and dyspnoea, rilpivirine/hydroxychloroquine with QT prolongation, bictegravir/ursodeoxycholic acid with rising viremia, and atorvastatin/fenofibrate with myalgia. Four DSIs between bictegravir and supplements containing divalent ions have been associated with increasing viremia. After medication review, alprazolam was replaced with zolpidem, rilpivirine-based HAART with bictegravir/emtricitabine/tenofovir alafenamide, and atorvastatin/fenofibrate with simvastatin/ezetimibe, and ursodeoxycholic acid and supplements were discontinued. These interventions were temporally associated with clinical improvement of interaction-related AEs and good virological control. Conclusions: This study highlights the importance of predicting DDIs/DSIs in PLWH undergoing polypharmacy. Medication review supports clinical decision-making, preventing avoidable adverse outcomes.

Pharmaceuticals doi: 10.3390/ph19091382

Authors: Sabrina Koribeche Sadjia Bertouche Nassila Sabba Naima Sahraoui Farah Djelti Faisal K. Alkholifi Rana M. Al-dossari Mohamed Said Kahaleras Mostefa Hani Yazid Chetbani Yacine Karmi Samia Daoudi-Hacini

Background/Objectives: Chronic inflammation sustained by cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) underlies many human diseases, while the cardiovascular liabilities of coxibs have renewed interest in plant-derived alternatives. This study characterised the volatile composition of Corymbia citriodora essential oil under two distillation regimes and identified constituents able to inhibit human COX-2 and iNOS. Methods: Oils obtained by steam distillation (SD) and microwave-assisted steam distillation (MSD) were profiled by GC–MS/MS. All identified constituents were screened with SASA Vina against human COX-2 (PDB: 5KIR, 3LN1), ovine COX-1 (4COX) and human iNOS (3E7G), with native-ligand re-docking validating each protocol (RMSD 0.39–0.84 Å). The lead compound underwent density functional theory (B3LYP/3-21G/CPCM), 100 ns all-atom molecular dynamics, MM/GBSA and MM/PBSA decomposition, and pkCSM ADMET prediction. Results: Sixty-three constituents were identified (99.85% SD; 99.97% MSD). MSD enriched oxygenated monoterpenes (93.23% versus 88.27%), citronellal rose from 38.24% to 48.41%. (Z,Z,Z)-1,5,9,9-Tetramethyl-1,4,7-cycloundecatriene ranked highest at COX-2 (−8.0 to −8.2 kcal/mol) and also bound COX-1 (−8.8 kcal/mol) and iNOS (−6.8 kcal/mol). DFT indicated high kinetic stability (ΔE = 6.12 eV; η = 3.06 eV) and purely non-covalent hydrophobic binding. The COX-2 complex remained stable over 100 ns (Cα RMSD 0.17 ± 0.02 nm), with MM/GBSA and MM/PBSA binding energies of −23.98 and −21.95 kcal/mol and Val523 as the principal hotspot. ADMET prediction returned 96.61% human intestinal absorption and no mutagenicity, hepatotoxicity or hERG I blockade. Conclusions: MSD offers a faster route to a citronellal-enriched oil, and C. citriodora hydrocarbon sesquiterpenes emerge as chemically stable, multi-target COX-2/iNOS scaffolds. Comparable binding at COX-1 indicates that isoform selectivity remains to be established; in vitro enzymatic and cell-based validation is therefore required.

Pharmaceuticals doi: 10.3390/ph19091381

Authors: Mihai Cristian Neagu Diana Haj Ali Emil Radu Iacob Andreea Smeu Roxana Stoicescu Călin Marius Popoiu Georgiana Boştinaru Ştefan Marcu Robert Ianoş Vlad Socoliuc Lucian Barbu Tudoran Elena-Alina Moacă

Background/Objectives: Magnetic iron oxide nanoparticles are investigated in cancer research; however, the direct biological effects of unloaded magnetic colloidal suspensions in renal cancer remain insufficiently characterized. This study compared two double-oleic-acid-coated formulations prepared from precursors: magnetite-based MCS 1 and maghemite-based MCS 2. Methods: The suspensions were characterized by vibrating-sample magnetometry, dynamic light scattering, bright-field scanning transmission electron microscopy, and energy-dispersive X-ray spectroscopy. Their effects on A704 renal adenocarcinoma cells were evaluated after 24 h of exposure to 1–5 µg/mL using MTT, neutral red uptake, JC-1, Hoechst 33342/MitoTracker Red CMXRos, acridine orange/propidium iodide staining, and a 7-day clonogenic assay. Results: MCS 2 showed higher volumetric saturation magnetization (1.541 vs. 1.059 Gs), a smaller Z-average hydrodynamic diameter (79.65 vs. 103.9 nm), and a more uniform volume-weighted distribution. Both formulations significantly reduced metabolic activity and neutral red uptake and induced mitochondrial depolarization, cell-death-associated morphological changes, and impaired clonogenic capacity. MCS 1 generally produced a moderate response with an apparent plateau, whereas MCS 2 showed a pronounced concentration-related effect at 4–5 µg/mL. At 5 µg/mL, MCS 2 reduced metabolic activity to approximately 49%, neutral red uptake to 50.04%, the JC-1 aggregate-to-monomer ratio to 46.02%, and colony formation to 31.44% of the control. Conclusions: The suspensions exhibited distinct physicochemical and biological profiles. MCS 2 produced greater effects at the highest concentrations, potentially related to its smaller hydrodynamic size and more uniform particle-size distribution. Further studies should establish tumor selectivity, cellular uptake, and the underlying molecular mechanisms.

Pharmaceuticals doi: 10.3390/ph19091380

Authors: Misa Kimura Yoichi Sakurada Yoshiko Fukuda Yumi Kotoda Kenji Kashiwagi

Purpose: This study aimed to evaluate how the temporal dynamics of polyp regression influence 24-month outcomes in eyes with polypoidal choroidal vasculopathy (PCV) treated with as-needed vascular endothelial growth factor (VEGF) inhibitors. Methods: Sixty-three eyes of 63 patients were included. All patients initially received three monthly intravitreal injections of either aflibercept (2 mg) or brolucizumab, followed by as-needed treatment for 24 months. The selection of a VEGF inhibitor depends on time. Indocyanine green angiography (ICGA) was performed at baseline, 3 months, and 12 months in all patients. Results: Among the 63 eyes, complete regression was observed in 39 eyes (61.9%) at 3 months. Of these 39 eyes, 29 eyes (46.0%) maintained complete regression at 12 months. Among the 24 eyes that did not show complete regression at 3 months, 5 eyes (20.8%) achieved complete regression at 12 months. Visual acuity improved over 24 months regardless of the timing of regression. Early regression at 3 months was associated with fewer additional injections during the first year. In contrast, regression at 12 months—rather than at 3 months—was more strongly associated with fewer additional injections during the second year. Conclusions: The most recent status of the polypoidal lesion was associated with the number of additional injections during the corresponding 12-month period, suggesting that ICGA-based assessment may be important for predicting treatment responsiveness.

Pharmaceuticals doi: 10.3390/ph19091379

Authors: Nick Zhang Marlys Kutach Lexis Hsieh Shelby Vexler Irene A. Chen

Antibiotic resistance of microorganisms threatens global health and agriculture. Antimicrobial peptides can display broad-spectrum activity, making them candidate alternatives to antibiotics. Here, ten antimicrobial peptides were tested against fungal pathogens, including Candida albicans, Cryptococcus neoformans, Aspergillus niger, and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive (Staphylococcus aureus) pathogens. Aurein 1.2, halictine-II, and macropin-I showed broad-spectrum activity. In particular, halictine-II showed high potency combined with low hemolytic activity, with the MIC values across species being 5 to 90-fold lower than the observed hemolytic concentration.

Pharmaceuticals doi: 10.3390/ph19091378

Authors: Janusz K. Rybakowski Cezary Mydlak Nadzieja Klimek Krzysztof Książek

Lithium remains the gold-standard maintenance treatment for bipolar disorder (BD) and is distinguished by its unique anti-suicidal and neuroprotective properties. Beyond its established psychiatric efficacy, growing evidence suggests that lithium may modulate fundamental mechanisms of biological aging. In parallel, cellular senescence has emerged as a central process linking oxidative stress, chronic inflammation, mitochondrial dysfunction, and impaired cellular resilience with neurodegeneration and psychiatric disease. Notably, BD is increasingly associated with features of accelerated aging, including telomere shortening, increased inflammatory burden, and structural brain changes consistent with premature brain aging. In this review, we discuss current evidence supporting lithium as a potential senostatic agent in the central nervous system. Experimental studies indicate that lithium attenuates several hallmarks of cellular senescence and promotes cellular resilience under conditions of oxidative, inflammatory, and genotoxic stress. These effects appear to involve coordinated modulation of neuroinflammatory signaling, mitochondrial function, oxidative stress responses, genomic stability, and neurotrophic pathways. Clinical findings further suggest that chronic lithium treatment may be associated with preserved telomere length and attenuated biological brain aging in BD. Collectively, the available data support a model in which lithium acts not only as a mood stabilizer but also as a broader regulator of aging-associated processes relevant to neuropsychiatric and neurodegenerative disorders. Although the senomodulatory effects of lithium appear context- and cell-type-dependent, its established clinical use and pleiotropic biological actions make it a promising candidate for translational senotherapeutic research.

Pharmaceuticals doi: 10.3390/ph19091377

Authors: Melinda Ildiko Mitranovici Raluca Moraru Septimiu Voidazan Florin Buicu Ion Petre Liviu Moraru

Natural compounds in topical gel formulations are recognized for their ability to enhance wound healing, particularly in the context of postoperative care, including that of cesarean sections. Background: Cesarean sections are among the most common surgical procedures globally, highlighting the importance of their successful healing. Complementary medicine aimed at pain relief and wound healing after a cesarean section has been investigated worldwide, and natural compounds are considered safe for topical use during pregnancy and breastfeeding. The aim of our study is to demonstrate the efficacy of the use of natural compounds in post-cesarean scar treatment. Materials and Methods: We compared Reparis, a topical gel entirely composed of natural compounds such as Centella asiatica and Aloe vera, with Cicatridine, a topical gel containing synthetic and natural compounds, such as almond oil, in post-cesarean wound treatment. We conducted an observational retrospective study in County Hospital Hunedoara between 26 February and 31 May 2026, evaluating specific measures that relate to the cosmetic results of scars after surgical procedures in our unit, with 50 patients using Reparis twice a day compared to the control group, which itself consisted of 50 patients using Cicatridine twice a day for 30 days. We employed the POSAS, an internationally validated questionnaire for evaluating patient and surgeon opinions regarding the appearance of a scar. Results: The most important difference favoring Reparis was observed after 6 months (p = 0.0001). In addition, Cicatridine exhibits a positive correlation with BMI; Reparis can be used in cases of higher BMI with good healing results. Conclusions: Natural compounds alone, such as Reparis, pose promising potential for wound healing, based on the findings of our observational study. The therapeutic properties of traditional medical plants have been verified but not always in controlled clinical trials. Our findings still require external validation.

Pharmaceuticals doi: 10.3390/ph19091376

Authors: Hayeong Jeon Yoo Jin Kim Geun Hee Seol

Background: Intracellular Ca2+ dysregulation and neurovascular dysfunction are increasingly recognized as important mechanisms underlying neurodegenerative disorders, including Alzheimer’s disease (AD). Restoration of pathological Ca2+ imbalance has therefore emerged as a potential therapeutic strategy. Although ginsenoside compound K (CK) has demonstrated neuroprotective and anti-inflammatory properties, its role in Ca2+ homeostasis remains unclear. Methods: SH-SY5Y neuronal cells, U373 astrocytes, BV2 microglia, bEND brain endothelial cells, and MOVAS vascular smooth muscle cells were exposed to trimethyltin chloride (TMT, 5 μM, 24 h). Cell viability, real-time cell confluence, intracellular Ca2+ influx, and endoplasmic reticulum (ER) Ca2+ store release were evaluated. Ca2+ dynamics were analyzed using Fura-2 fluorescence, and signaling associated with CK-mediated Ca2+ regulation was investigated using pharmacological inhibitors. Orai1 protein expression was evaluated by Western blot analysis in SH-SY5Y and U373 cells. Results: TMT increased store-operated Ca2+ entry (SOCE)-mediated Ca2+ influx in SH-SY5Y neuronal cells without significant cytotoxicity. In contrast, TMT reduced both Ca2+ influx and cell viability in U373 astrocytes, BV2 microglia, and MOVAS cells, whereas bEND cells showed minimal changes. CK restored abnormal Ca2+ responses in a cell type-specific manner, reducing elevated Ca2+ influx in neuronal cells while restoring suppressed Ca2+ signaling in astrocytes. Consistent with these functional findings, Orai1 protein expression was increased by TMT in SH-SY5Y cells and attenuated by CK, whereas TMT reduced Orai1 expression in U373 cells, which was restored by CK. Pharmacological analyses suggested involvement of PKA- and LTCC-associated signaling in neuronal cells and PLC- and PLD-associated signaling in astrocytes. In both cell types, CK-associated Ca2+ responses were sensitive to NAC and the SOCE inhibitor BTP2, supporting ROS- and SOCE-associated mechanisms. Additional controls showed no significant inhibitor effects under control or CK-only conditions, strengthening the pharmacological interpretation of responses under TMT-containing conditions. Conclusions: TMT-induced Ca2+ dysregulation differed markedly by cell type. CK restored disrupted Ca2+ homeostasis in association with distinct pharmacological signaling profiles in neuronal and astrocytic cells. These findings suggest that CK may function as a cell type-specific Ca2+ homeostatic regulator under neurotoxic stress conditions and highlight the importance of differential Ca2+ regulation in neurodegenerative disease models.

Pharmaceuticals doi: 10.3390/ph19091375

Authors: Wanying Xie Sarah Beishan Tai Chang-Tong Yang Sidney W. K. Yu Young Soon Tay Ruban Poopalalingam Darren Liang-Khai Koh Kerry Riffel Wenping Li Chih-Liang Chin Asad abu Bakar Ali Darren Wan-Teck Lim David Chee Eng Ng

Background: Metabotropic glutamate receptor subtype 5 (mGluR5) is a promising therapeutic target for a range of neuropsychiatric disorders, and [18F]FPEB is a PET radioligand developed for imaging central mGluR5 receptor availability. This study aimed to determine the biodistribution, kinetic characteristics, and radiation dosimetry of [18F]FPEB in healthy Asian subjects and to evaluate the feasibility of reference tissue models as alternatives to conventional blood-based kinetic modeling. Methods: Five healthy Asian male volunteers underwent whole-body and dynamic brain PET imaging following administration of [18F]FPEB. Radiation dosimetry was estimated using OLINDA/EXM software. Tracer kinetics were analyzed using both blood-based compartmental models and reference tissue models to derive regional binding potentials (BPND) and volumes of distribution. Model performance was assessed using the Akaike Information Criterion (AIC). Results: The mean whole-body effective radiation dose was 0.703 mSv/mCi, with the gallbladder identified as the critical organ receiving the highest absorbed dose (5.63 mGy/mCi). Brain time–activity curves demonstrated the highest tracer uptake in the nucleus accumbens, followed by the cingulate gyrus, putamen, and insula, with moderate uptake in the thalamus and globus pallidus and the lowest uptake in the brainstem and cerebellum. Using the simplified reference tissue models (SRTM/SRTM2), the highest BPND values were observed in the nucleus accumbens, followed by the cingulate gyrus, putamen, and insula. In contrast, two-tissue compartment models (2TCM/2TCM-C) showed the highest BPND values in the hippocampus, followed by the amygdala, nucleus accumbens, and caudate nucleus. Although differences were observed in the absolute BPND values and regional ranking, both modeling approaches identified similar brain regions with high mGluR5 binding. Conclusions: [18F]FPEB demonstrated a favorable radiation dosimetry profile and supports the feasibility of reference tissue models for BPND estimation in healthy Asian subjects. These findings provide preliminary baseline data, although validation in larger cohorts is needed to confirm the pharmacokinetic characteristics and modeling approach.

Pharmaceuticals doi: 10.3390/ph19091374

Authors: Qinxiu Zhang Shouwei Sun Miaomiao Lu Runxin Qin Junxuan Ren Lianjie Yao Dianlong Jia Jinjie Chang Xiaohong Chu Rui Wang Fang Liu Jun Li

Background: Pulmonary fibrosis (PF) is a progressive lung disease with limited therapies. Oral Pirfenidone (PFD), an approved anti-fibrotic, shows poor lung bioavailability and significant toxicity. S-Allylmercapto-N-acetylcysteine (ASSNAC) has been demonstrated to possess significant anti-inflammatory and antioxidant properties. Methods: In this study, dry powder inhaler formulations (DPIs) co-encapsulating ASSNAC and PFD in liposomes are reported. The formulation was prepared via spray freeze-drying (SFD) using L-leucine as both a cryoprotectant and a surface morphology modifier. The interfacial enrichment of L-leucine during atomization and freezing created a hydrophobic, corrugated surface that lowered particle surface energy, prevented liquid/solid bridge formation, and reduced hygroscopicity. Results: The resulting liposomal powders exhibited optimal aerosol performance: fine particle fraction (FPF) of 61.08% ± 2.45% and mass median aerodynamic diameter (MMAD) of 2.22 ± 0.11 µm. Cellular studies demonstrated that dual-loaded liposomes were efficiently taken up by Beas-2B and HFL-1 cells, with no cytotoxicity (cell viability > 90%) and negligible hemolysis. In the TGF-β1-induced in vitro fibrosis model, the combined treatment exerted prominent therapeutic effects. This regimen preserved normal epithelial morphology, accelerated wound repair, suppressed fibroblast invasiveness, and lowered the expression levels of Collagen I, Collagen III, and α-SMA. In bleomycin-induced pulmonary fibrosis rats, inhaled ASSNAC + PFD liposomal powder markedly relieved lung tissue injury and collagen accumulation. It restored redox balance and lowered pulmonary TGF-β1, hydroxyproline and collagen III levels. The formulation blocked TGF-β1 signaling and fibrotic gene expression. Conclusions: Pulmonary administration avoids oral pirfenidone-induced liver and stomach toxicity. Collectively, the ASSNAC + PFD co-encapsulated liposomal dry powder produced by SFD represents a safe and efficacious therapeutic strategy for PF.

Pharmaceuticals doi: 10.3390/ph19091373

Authors: Seung-Hyeon Ahn Jihyun Shin Hwayoung Na Hong Kyu Lee Kyung-Chul Choi

Background/Objectives: Gastric cancer remains a leading cause of cancer-related mortality worldwide, highlighting the need for novel therapeutic strategies. Drug repositioning offers a cost-effective approach by identifying new therapeutic applications for clinically approved drugs. Fluphenazine (FPZ), a dopamine receptor D2 antagonist used as an antipsychotic drug, has demonstrated anticancer activity; however, its effects on gastric cancer remain unclear. Methods: This study investigated the anticancer effects of FPZ alone and in combination with cisplatin (DDP) in gastric cancer. Results: In MKN-45 cells, viability was 100%, 101.16%, 61.26%, and 53.12% in the control, FPZ, DDP, and combination groups, respectively; the corresponding values in AGS cells were 100%, 84.95%, 89.94%, and 73.94%. FPZ also inhibited migration and induced apoptosis, while combined treatment increased cytosolic and mitochondrial reactive oxygen species levels and mitochondrial stress. Furthermore, FPZ reduced IL-6 and IL-8 expression and attenuated DDP-induced inflammatory responses. In the mouse xenograft model, the tumor volumes were 1233.65, 1115.75, 768.88, and 309.44 mm3 in the control, FPZ, DDP, and combination groups, respectively. Conclusions: These findings support the potential of FPZ as a repurposed anticancer agent for gastric cancer, particularly in combination with DDP.

Pharmaceuticals doi: 10.3390/ph19091372

Authors: Lihao Wang Wuyou Gao Yuesong Xiao Ting Yang Jingwei Wang Yanping Sun

Background: Safe treatment options for ulcerative colitis, a chronic relapsing inflammatory bowel disease, remain limited, and the bioactive fraction of Rosa davurica Pall. root associated with its protective activity against Ulcerative colitis (UC) has not been characterized. Methods: This study evaluated the protective effects of the total root aqueous extract (WRP) and two process-defined fractions prepared by ethanol precipitation: the supernatant fraction (SRP) and crude polysaccharide precipitate fraction (PRP). These fractions were administered concurrently with Dextran sulfate sodium (DSS) during acute colitis induction in mice. Assessments included disease activity index, colon histopathology, intestinal barrier proteins, RT-qPCR analysis, fecal metabolomics, 16S rRNA sequencing, and UHPLC-MS/MS. Results: Compared to PRP, SRP significantly alleviated DSS-induced weight loss, colon shortening, perianal bleeding, and histological injury. SRP also reduced the mRNA levels of TNF-α, IL-6, IL-1β, MPO, NF-κB, NLRP3, GSDMD, and IL-18. SRP treatment increased Claudin-1 and ZO-1 protein expression, whereas Occludin expression showed no significant change. SRP treatment was associated with shifts in fecal metabolic profiles and gut microbiota composition in DSS-treated mice, including a change in the Firmicutes/Bacteroidota ratio toward the CON group pattern. UHPLC–MS/MS profiling resulted in the tentative annotation of 68 nonredundant constituents in SRP. Conclusions: Under the present experimental conditions, SRP demonstrated protective activity in DSS-induced acute colitis and represents a potentially important bioactive fraction of the R. davurica root aqueous extract. Its protective effects were accompanied by reduced inflammatory gene expression, partial enhancement of barrier-associated protein expression, and alterations in gut microbiota composition and fecal metabolic profiles.

Pharmaceuticals doi: 10.3390/ph19091371

Authors: Laura Radoš Sara Matulić Čubranić Marin Golčić Iva Skočilić Ivana Mikolašević Andrej Belančić

Background: Pancreatic adenocarcinoma (PDAC) remains one of the most lethal malignancies, with limited survival gains despite advances in systemic therapy. However, rapid expansion of biomarker-directed therapies, immunotherapy, and novel treatment modalities has created an increasingly complex clinical trial landscape. We aimed to characterize contemporary PDAC clinical development and identify emerging therapeutic trends. Methods: We performed a narrative review with a scoping approach of active PDAC clinical trials registered on ClinicalTrials.gov. Eligible studies were initiated between January 1, 2021, and February 16, 2026, and included recruiting, active, or not-yet-recruiting phase I–III trials. Data extracted included trial phase, disease setting, therapeutic strategy, endpoints, enrollment, sponsorship, and late-phase development. Results: A total of 355 interventional trials were included. Most trials were phase I, phase I/II, or phase II trials, with fewer than 10% being evaluated in phase II/III or phase III development. Advanced or metastatic disease was the predominant setting. Chemotherapy remained the most frequently incorporated treatment modality, while molecularly targeted therapies were evaluated in 167 trials. KRAS/RAS-directed approaches represented the largest targeted subgroup, although only daraxonrasib and setidegrasib reached phase III evaluation. Immunotherapy was evaluated in 139 trials, although only a limited number progressed to late-phase development, reflecting the immune-resistant biology of pancreatic adenocarcinoma. Additional areas of active investigation included Claudin 18.2-targeted therapies, MTAP-associated approaches, homologous recombination deficiency-directed strategies, CD73 inhibition, radiotherapy, local interventions, surgery-focused optimization, imaging-guided approaches, and supportive-care interventions. Industry organizations were listed as the lead sponsor for approximately half of all studies, while non-commercial organizations supported most remaining trials. Conclusions: The contemporary PDAC clinical trial landscape is characterized by broad therapeutic diversification but limited late-phase maturity. Chemotherapy remains the dominant treatment backbone, whereas KRAS/RAS-directed therapies have emerged as the most advanced precision oncology strategy. Future progress will likely depend on successful integration of biomarker-selected therapies with established multidisciplinary treatment approaches, including optimized systemic therapy, local-control strategies, and supportive care.

Pharmaceuticals doi: 10.3390/ph19091370

Authors: Daniel Miron Brie Cristian Mornoș Alexandru Tîrziu Roxana Popescu Alina Diduța Brie

Background: Chronic coronary syndrome (CCS) is a major source of morbidity and healthcare utilization, and a substantial proportion of patients continue to experience angina despite contemporary revascularization and guideline-directed medical therapy. Trimetazidine, a metabolic anti-ischemic agent that does not affect heart rate or blood pressure, has been used for decades as a non-hemodynamic add-on. Its position in international guidelines, however, has shifted. We aimed to synthesize current evidence on the clinical efficacy, safety, and guideline status of trimetazidine in CCS and coronary artery disease (CAD) and to delineate clinical circumstances in which a symptom-directed trial may be considered. Methods: We performed a structured narrative review of randomized controlled trials, network and conventional meta-analyses, and large observational studies indexed in PubMed/MEDLINE, Embase, and the Cochrane Library between January 1990 and March 2026. Reference lists of eligible articles, international clinical practice guidelines, and regulatory communications from European and national agencies were also examined. Results: Across pooled analyses, trimetazidine modestly but reproducibly improved exercise tolerance and reduced angina frequency and short-acting nitrate use, with efficacy comparable to other non–heart-rate-lowering antianginal agents. Improvements in left ventricular function were reported in small, single-center studies in ischemic cardiomyopathy and selected high-risk subgroups; however, these findings have not been validated in contemporary large-scale trials, and their clinical significance against modern guideline-directed therapy is unknown. The ATPCI trial confirmed safety but showed no reduction in major adverse cardiovascular events after percutaneous coronary intervention. Rare extrapyramidal effects prompted the 2012 European Medicines Agency referral and several prescribing restrictions. Conclusions: Trimetazidine remains a reasonable symptom-directed adjunct for residual angina in carefully selected CCS patients, particularly those in whom further hemodynamic up-titration is limited or poorly tolerated. The 2024 ESC class IIb recommendation may partly reflect a broader emphasis on prognostic benefit, although the neutral post-PCI outcome evidence and regulatory safety restrictions also support a more cautious position.

Pharmaceuticals doi: 10.3390/ph19091369

Authors: Wael Jalloul Cristina Mariana Uritu Despina Jalloul Vlad Ghizdovat Andreia Vranceanu Ciobanu Bogdan Ionel Tamba Cipriana Stefanescu Irena Cristina Grierosu

Parkinsonian syndromes comprise overlapping neurodegenerative and non-degenerative disorders, making aetiological diagnosis difficult, while existing procedural guidance does not fully integrate tracer-specific biology with clinical decision-making, multimodal strategies, and emerging quantitative and pathology-directed biomarkers. This review synthesises evidence on the molecular targets, radiopharmaceutical characteristics, biological interpretation, and clinical applications of dopaminergic single-photon emission computed tomography (SPECT) and positron emission tomography (PET), focusing on the dopamine transporter (DAT), aromatic L-amino acid decarboxylase (AADC), vesicular monoamine transporter type 2 (VMAT2), dopamine D2/D3 receptors, differential diagnosis, semiquantification, kinetic modelling, and artificial intelligence-assisted interpretation. The evidence confirms that DAT SPECT with [123I]ioflupane ([123I]FP-CIT) remains the most established method for demonstrating or arguing against presynaptic nigrostriatal dysfunction, but an abnormal result cannot establish aetiology, and a normal result can redirect evaluation towards non-degenerative mimics; target-specific PET provides complementary biological information, although availability, standardisation, and prospective validation remain limiting. By additionally integrating genetic and prodromal applications, question-driven multimodal imaging, emerging acquisition approaches, α-synuclein imaging and seed amplification assays, and contemporary biological staging frameworks, this review extends procedural guidance and positions dopaminergic imaging as a targeted functional biomarker selected according to the unresolved clinical question rather than as a stand-alone disease label.

Pharmaceuticals doi: 10.3390/ph19091368

Authors: Xin Zeng Wen-Feng Du Wen-Hao Yin Jun-Yao Zhu Yu-Bin Yang Wei-Jun Guo Wen-Liang Li Hao Gong Zi-Zhong Yang Yi Li

Background: Orientothele washanensis is a venomous spider with considerable ecological and scientific importance. Its venom, characterized by complex composition and ease of collection, serves as a valuable resource for the discovery of natural peptide drugs. Conventional wet-lab screening methods are limited by rigorous experimental conditions, high resource consumption, and long research cycles, which hinder the efficient identification of functional peptides from the venom gland transcriptome of this spider. Methods: To address these technical bottlenecks, this study developed a novel deep learning model named PepPI-DRN for peptide-protein interaction prediction. The model integrated a residual equivariant graph neural network, a residual 1-dimensional convolutional neural network, and a dual-modal attention mechanism by leveraging both sequence and structural features of peptides and proteins. Results: Results on an independent test set indicated that PepPI-DRN achieved the competitive or superior performance compared with state-of-the-art methods on multiple key evaluation metrics. Candidate peptides with high interaction probabilities against targets were obtained from the venom gland transcriptome of Orientothele washanensis. Furthermore, lead peptides with high binding strength and good structural stability were identified from candidate peptides by molecular docking, and molecular dynamics simulation. Conclusions: These results showed that the pipeline with PepPI-DRN, molecular docking and molecular dynamics simulation enabled efficient and reliable identification of lead peptides from the venom gland transcriptome of Orientothele washanensis, providing a robust and effective strategy for the discovery and development of natural peptide drugs from the spider venom.

Pharmaceuticals doi: 10.3390/ph19091367

Authors: Xi-Yuan Feng Meng Gao Fei-Long Liu Ming-Ze Li Xiao-Li Cui Meng-Yang Wang Zhi-Hong Zhang He Li Chun-Mei Wang Jing-Hui Sun

Objectives: This study focuses on exploring the mechanism by which Schisandrin B (Sch B) regulates metabolic dysfunction-associated steatotic liver disease (MASLD) mice induced by a high-fat methionine–choline-deficient (MCD) diet through the activation of peroxisome proliferator-activated receptor γ (PPARγ). Methods: Male C57BL/6 mice were fed a high-fat MCD diet for 8 weeks to establish a mouse MASLD model, and the effects of Sch B on MASLD and the mechanisms were investigated. PPARγ overexpression (OE) was induced by adeno-associated virus (AAV) administration via intrahepatic portal vein injection in mice, and a negative control (NC-OE) was also established. Body weight; wet liver weight; hepatic index; serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β); and hepatic triglyceride (TG) levels were measured in the mice. The histopathology and lipid deposition were observed by hematoxylin and eosin (H&E) staining and Oil Red O staining, while the fibrosis was assessed using Masson staining. Western blot was employed to detect the expression levels of PPARγ, sterol regulatory element-binding protein 1c (SREBP-1c), carnitine palmitoyltransferase 1A (CPT1A), transforming growth factor β1 (TGF-β1), α-smooth muscle actin (α-SMA), collagen type I (collagen I), Smad family members 2/3 (Smad2/3), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38), and extracellular signal-regulated kinase 1/2 (ERK1/2), along with the phosphorylation activation status of these kinases. Results: It was confirmed that Sch B caused effects similar to those induced by PPARγ overexpression, reducing the hepatic index, AST, and ALT levels while alleviating lipid accumulation and fibrosis; and upregulating PPARγ and CPT1A while inhibiting SREBP-1c; and the phosphorylation of the TGF-β/Smad and MAPK pathways were involved in the mechanisms. Conclusions: Sch B can alleviate high-fat MCD-induced MASLD by activating PPARγ in mice.

Pharmaceuticals doi: 10.3390/ph19091366

Authors: Samridhi Kurl Neeraj Mittal Rajwinder Kaur Balakumar Chandrasekaran Gurpreet Kaur

Background/Objectives: Psychobiotics are live microorganisms that, when administered in appropriate amounts, can enhance mental health. Through the production of neuroactive metabolites, regulation of immune responses, and interactions along the gut–brain axis, they modulate host neurophysiology. However, significant strain-specific variability exists in their metabolic capabilities, colonization potential, and functional interactions, necessitating a systematic approach for psychobiotic strain selection. Methods: In the present study, an integrated in silico systems biology approach combining comparative genomics, genome-scale metabolic modeling, network pharmacology, molecular docking, molecular dynamics, and ADMET analysis was employed to computationally evaluate the therapeutic potential and molecular interactions of a quercetin-psychobiotic co-delivery system for managing depression by modulating the gut–brain axis. Results: Comparative genomic analysis of candidate psychobiotic strains from the genera Bifidobacterium and Lactobacillus predicted the presence of key functional genes involved in neuroactive metabolite production, immune modulation, oxidative stress tolerance, and adhesion. The presence of acetate-producing pathways alongside the absence of classical butyrate biosynthesis genes indicated an indirect contribution to short-chain fatty acid production via microbial cross-feeding interactions. Genome-scale metabolic Modeling under the conditions of quercetin supplementation predicted a significant inter-strain variation in terms of predicted biomass flux, nutrient utilization, and metabolic adaptability. Results of molecular docking showed that the binding energies were in the range of −5.854 to −9.489 kcal/mol. Conclusions: Collectively, these in silico findings provided a computational basis for the selection of psychobiotic strains and predicted that a combination of psychobiotics and quercetin could present a promising strategy for the modulation of the gut–brain axis.

Pharmaceuticals doi: 10.3390/ph19091365

Authors: Yiğit Erkmen Zekiye Ceren Arıtuluk Aydın Engin Koçak Hasya Nazlı Gök Emirhan Nemutlu Merve Yüzbaşıoğlu Baran

Background/Objectives: Berberis species are recognized for their ethnomedicinal importance and phytochemical richness associated with antidiabetic activity. Although B. crataegina is widely distributed in the Turkish flora and has a history of traditional use, its phytochemical properties and biological activities remain less extensively investigated than those of other Berberis species. This study aimed to comparatively evaluate the phytochemical profiles, in vitro antidiabetic potential, and cytoprotective effects of extracts prepared from different parts of B. crataegina in a high-glucose-induced SH-SY5Y cell-based model of diabetic neuropathy. Methods: Crude hydroethanolic extracts (70% ethanol) were prepared from the leaves, flowers, shoots, roots, and fresh fruits of B. crataegina collected from Kızılcahamam, Ankara, Türkiye. The total phenolic and flavonoid contents, antioxidant capacities and α-glucosidase inhibitory activities of leaf, flower, shoot, root, and fruit extracts were determined. The cytoprotective effects of the crude extracts were evaluated in a high-glucose-induced SH-SY5Y cell model. The metabolomic profiles of the extracts were comparatively analyzed using liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). The most active fruit extract was fractionated by reversed-phase vacuum liquid chromatography (RP-VLC), and the phytochemical contents and α-glucosidase inhibitory activities of the fractions were evaluated. The major phenolic compounds in the most active fruit extract were quantitatively determined using high-performance liquid chromatography–diode array detection (HPLC-DAD). The relationships between metabolites and biological activities were investigated through correlation analysis. Results: The fruit extract was distinguished by its α-glucosidase inhibitory activity, total phenolic content, antioxidant capacity, and cytoprotective effect in the high-glucose-induced SH-SY5Y cell model. It showed no marked cytotoxicity at concentrations ranging from 25 to 400 µg/mL and, at 25 µg/mL, attenuated the high-glucose-induced reduction in cell viability, restoring viability to a level close to that of the normal control. Untargeted metabolomic analysis identified 191 metabolites detected in at least two crude extracts, while the metabolites detected exclusively in the fruit extract were predominantly flavonoids and flavonoid glycosides. Fractionation showed that α-glucosidase inhibitory activity was mainly concentrated in the medium-polarity fractions; however, none of the fractions reached the activity level of the crude extract. Chlorogenic acid, rutin, caffeic acid, protocatechuic acid, quercetin, and quercetin-3-O-glucoside were detected and quantified in the fruit extract by HPLC-DAD. Correlation analysis indicated that the observed biological activities may be associated with phenolic acids, flavonoids, and other polyphenolic metabolites. Conclusions: The fruit extract of B. crataegina showed promising α-glucosidase inhibitory activity and cytoprotective effects in the high-glucose-induced SH-SY5Y cell model. The findings suggest that these effects may arise from the combined contribution of multiple constituents within a polyphenolic matrix rather than from a single compound; however, synergistic interactions were not experimentally demonstrated. These results support further evaluation of the fruit extract through mechanistic, in vivo, and standardization studies.

Pharmaceuticals doi: 10.3390/ph19091364

Authors: Sonu Kamal Y. Thajudeen Saad Ali Alshehri Mohammed Muqtader Ahmed Mayur Porwal Sagar Verma

Background: The current study uses a computational technique to investigate synthesized thiazine compounds that exhibit potential interactions with the GABAA receptor. A series of novel thiazine derivatives was synthesized via the cyclization reaction of chalcone with thiourea. Methods: For the synthesized compounds, spectroscopic characterization, namely FT-IR, 1H NMR, 13C NMR, and mass spectrometry, was carried out. In addition, the AutoDock Vina 4.2 software was applied to carry out an in silico docking study. Results: The synthesized compounds C3 and C6 exhibited comparable predicted docking scores (−9.0 and −9.1 kcal/mol, respectively), with favorable predicted interactions at the GABAA receptor binding site. Furthermore, in silico ADMET analysis provided insights into the drug-likeness and pharmacokinetic profiles of the synthesized compounds; however, potential safety liabilities, including Ames positivity, hERG II liability, and hepatotoxicity, were predicted for some derivatives. Conclusions: It was determined that the synthesized compounds’ various physiologically significant and physiologically relevant parameters fell within the range of Lipinski’s rule of five. The synthesized compounds (C1–C10) were tested for anxiolytic activity by the elevated plus maze test, and compound C6 was found to be the most potent. Additionally, the in vitro GABA-AT enzyme activity assay showed that compound C6 had the highest inhibitory potential, with an IC50 of 13.29 ± 1.622 µM.

Pharmaceuticals doi: 10.3390/ph19091363

Authors: Chunlai Feng Qiuqi Feng Shengnan She Ruojing Yang Mengru Li Lu Gong Mengjie Rui

Background/Objectives: Breast cancer exhibits substantial molecular heterogeneity, resulting in diverse therapeutic vulnerabilities and limiting the efficacy of single-agent therapies. Although multi-target strategies may offer improved therapeutic benefit, the systematic identification of synergistic higher-order target combinations remains challenging. This study aimed to identify and validate effective higher-order target combinations for heterogeneous breast cancer. Methods: DeepMDS, a previously developed deep learning-based multi-compound synergy prediction model, was used to prioritize candidate target combinations for breast cancer. Exhaustive two-target and three-target combinations were ranked in the gene-expression contexts of ER-positive luminal-like MCF-7 and triple-negative MDA-MB-231 breast cancer cells. The top-ranked target combinations were evaluated using single, pairwise, and triple small interfering RNA (siRNA) perturbations. Representative inhibitors were subsequently assessed in fixed-ratio combinations in MCF-7, MDA-MB-231, and 4T1 cells and in a 4T1 syngeneic mouse experiment. Results: BIRC5-NAMPT-TOP1 ranked first in both cell lines. Triple siRNA co-transfection targeting BIRC5, NAMPT, and TOP1 produced the strongest antiproliferative effects, with inhibition rates of 62.94% in MCF-7 cells and 55.62% in MDA-MB-231 cells. The corresponding inhibitors, LQZ-7I, FK866, and topotecan, exhibited synergistic antiproliferative activity at multiple molar ratios, with combination index values below 1. The optimized 2.5:10:1 molar ratio showed strong synergy in MCF-7, MDA-MB-231, and 4T1 cells, with combination index values of 0.26, 0.19, and 0.15, respectively. In tumor-bearing mice model, the triple-inhibitor regimen achieved a tumor inhibition rate of 62.05%. Topotecan-containing groups showed hematological alterations, whereas no statistically detectable elevations were observed in the measured terminal serum hepatic or renal biomarkers. Conclusions: The BIRC5-NAMPT-TOP1 combination showed reproducible phenotypic activity in the tested genetic and pharmacological models. These findings support the use of DeepMDS as a hypothesis-generation tool for higher-order target prioritization.

Pharmaceuticals doi: 10.3390/ph19091362

Authors: Qin Liu Huiyi Peng Min Su Zhoujin Tan Maijiao Peng

Background: Shenling Baizhu Powder (SLBZP) is a traditional Chinese medicine formula used to treat diarrhea. This study investigated its therapeutic effects and potential mechanisms in mice with fatigue- and high-fat diet-induced diarrhea, focusing on gut microbiota, bile acid metabolism, and farnesoid X receptor (FXR)-related signaling. Methods: Male Kunming mice were used to establish a model of diarrhea, followed by treatment with SLBZP. Fecal moisture content, serum levels of diamine oxidase and liver enzymes, and intestinal and hepatic histopathological changes were assessed. Small-intestinal microbiota composition, bile acid profiles in colonic contents, and FXR-related signaling in the ileum and liver were further analyzed. Results: SLBZP significantly reduced fecal moisture content. It also modulated the small-intestinal microbiota, with a significant increase in Faecalibaculum. In colonic contents, SLBZP reduced total bile acid levels, increased the secondary-to-primary bile acid ratio, and altered multiple bile acid species. These changes were accompanied by increased ileal FXR expression and changes in downstream regulators involved in hepatic bile acid synthesis. Correlation analysis further showed associations of selected bacterial taxa and FXR protein expression with colonic bile acid profiles. Conclusions: SLBZP alleviates diarrhea induced by fatigue combined with a high-fat diet in mice. Its effects may involve coordinated modulation of the gut microbiota, bile acid homeostasis, and FXR-related signaling, providing experimental support for the pharmacological basis of this traditional formula in diet-related diarrhea.

Pharmaceuticals doi: 10.3390/ph19091361

Authors: Jayoung Oh Ji Ho Choi

Background: Obstructive sleep apnea (OSA) remains under-treated because continuous positive airway pressure, though efficacious, is frequently limited by poor tolerance and adherence. Long considered a purely anatomical disease refractory to drugs, OSA is now understood as the shared endpoint of four measurable endotypes—a collapsible upper airway, poor pharyngeal dilator-muscle responsiveness, high loop gain, and a low arousal threshold—each a distinct pharmacological target. This review synthesizes the mechanistic rationale and clinical evidence for targeting them. Methods: In this narrative review, we searched PubMed/MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials (January 2000 to February 2026) for randomized trials, meta-analyses, and mechanistic endotyping studies, appraising the evidence narratively. Results: Tirzepatide became the first drug approved for OSA, acting through weight loss to reduce obesity-related mechanical loading, and the noradrenergic–antimuscarinic combination AD109 met its primary endpoint in a pivotal phase 3 trial by targeting dilator-muscle activity independently of weight. Both, however, used AHI-based endpoints in clinically defined rather than endotype-selected populations; AD109’s placebo-corrected effect was modest (−4.0 events/h), and 21.2% versus 3.1% of participants discontinued for adverse events. They are therefore best described as mechanism-informed rather than endotype-validated. Carbonic anhydrase inhibition with acetazolamide or sultiame and selected sedating agents address ventilatory instability and a low arousal threshold, while solriamfetol and pitolisant relieve residual sleepiness. Conclusions: Mechanism-informed pharmacotherapy for OSA is now clinically credible, but scalable bedside endotyping, attention to tolerability, and trait-stratified trials are needed before treatment can be called endotype-guided.

Pharmaceuticals doi: 10.3390/ph19091360

Authors: Dongsheng Wei Han Li Mei Zhao Mai Liu Yongyue Liu Menglan Zhao Huiming Cao Xiaoqing Zhang

Background: Pathological cardiac remodeling is a major driver of heart failure progression and is closely associated with fibroblast activation, myocardial inflammation, oxidative injury, and impaired autophagic homeostasis. Epidermal growth factor receptor (EGFR) signaling participates in adverse remodeling through activation of MAPK pathways, but the relationship between pharmacologically modulated EGFR turnover and cardiac remodeling remains insufficiently defined. Methods: This study investigated the protective effects of cycloastragenol (CAG), a triterpenoid sapogenin derived from Astragalus membranaceus, in Angiotensin II (Ang II)-induced cardiac remodeling and examined changes in EGFR turnover and associated MAPK signaling and autophagy-related processes. Circulating soluble EGFR was associated with heart failure severity after adjustment for age and sex. Separately, an independent exploratory transcriptomic analysis of left ventricular samples from GSE161472 showed that myocardial EGFR expression distinguished HFrEF from non-failing samples with an AUC of 0.870. Male C57BL/6 mice subjected to continuous Ang II infusion at 500 ng/kg/min for 28 days were used to establish the in vivo cardiac remodeling model and evaluate the effects of CAG on cardiac function, myocardial injury, fibrosis, inflammation, oxidative stress, apoptosis, and autophagy-related changes, with enalapril used as a positive control. Primary cardiac fibroblasts were further used to assess proliferation, migration, collagen production, EGFR membrane accumulation, lysosomal trafficking, MAPK signaling, and autophagic flux. Circulating soluble EGFR increased with heart failure severity and was associated with BNP elevation, ventricular dilation, and reduced ejection fraction. Results: CAG improved cardiac performance, reduced myocardial injury markers, alleviated fibrosis and hypertrophy, and suppressed inflammatory and oxidative responses in Ang II-treated mice. In cardiac fibroblasts, CAG reduced sustained EGFR membrane accumulation, increased EGFR recovery in pan-ubiquitin immunoprecipitates and lysosomal localization, and accelerated EGFR protein turnover, consistent with a predominantly lysosome-associated pathway. The K716R mutation attenuated CAG-associated changes in EGFR turnover, MAPK phosphorylation, and autophagic flux. Conclusions: These findings suggest that CAG attenuates Ang II-induced cardiac remodeling and is associated with enhanced EGFR lysosomal turnover, accompanied by changes in MAPK signaling and autophagy-related processes.

Pharmaceuticals doi: 10.3390/ph19091359

Authors: Enikő Rauch Csilla Ari Dominic P. D’Agostino Zsolt Kovács

Background/Objectives: Inflammatory processes, including elevated pro-inflammatory cytokines such as interleukin 1β (IL-1β), have been implicated in the pathophysiology of anxiety disorders. The ketone body β-hydroxybutyrate (βHB) was shown to mitigate inflammatory processes induced by lipopolysaccharide (LPS) administration. It was also suggested that exogenous ketone supplements, such as ketone esters (KEs) and ketone salts (KSs), can decrease anxiogenic effects through increased blood ketone levels in the body (ketosis) in preclinical animal models. Methods: Thus, we investigated whether standard rodent food supplemented with KEKS (KEKS food: mix of a KE R/S-1,3-butanediol-acetoacetate diester and a KS Na+- and Ca2+- R/S-βHB salt in 1:1 ratio) for 7 days can alleviate the intraperitoneal (i.p.) 1 mg/kg LPS-generated anxiogenic effect in female Wistar Albino Glaxo/Rijswijk (WAG/Rij) rats. For the evaluation of anxiogenic effects, the light–dark box (LDB) test was used. Results: KEKS food significantly increased the blood R-β-hydroxybutyrate (R-βHB) level and decreased the blood glucose level compared to the baseline. Moreover, the LPS significantly (i) decreased the time spent in the light compartment, the number of chamber transitions, the number of re-entries to the light compartment and the number of rearing in the light compartment, and also (ii) increased the time of latency to the first re-entry to the light compartment compared to control. Nevertheless, KEKS food treatment abolished LPS-generated anxiety-like behavior. Conclusions: The current study demonstrated that a 7-day KEKS-supplemented diet can increase blood R-βHB levels and abolish acute LPS administration-evoked increases in anxiety levels in female WAG/Rij rats. Moreover, our results support the important role of neuroinflammation in pathological processes resulting in anxiety. Thus, these results suggest that exogenous ketone supplement-generated ketosis may be a promising therapeutic tool in the treatment of inflammatory process-mediated diseases, such as anxiety disorders.

Pharmaceuticals doi: 10.3390/ph19091358

Authors: Xiao-Hui Chi Ming-Feng Liao Ya-Qun Zhou

Although the underlying mechanisms of renal IRI have been extensively studied, the corresponding effective treatments are still lacking. HMGB1, an important nuclear factor that is secreted outside cells when experiencing stress conditions, acts as a DAMP and exerts crucial effects on renal IRI. Many studies have suggested that the effect of HMGB1 on kidney damage is mediated mainly through the interaction of HMGB1 with pattern recognition receptors such as TLR4 and RAGE, which then results in the aggravation of local inflammatory response, increased infiltration of leukocytes, and finally renal tubular damage. Preclinical studies using animal models have demonstrated that inhibition of HMGB1 and downstream signal pathways can attenuate renal injury. This review critically evaluates HMGB1 in renal IRI across redox state, subcellular localization, temporal and cell-specific release, receptor usage, integrated stress pathways, autophagy, and regulated cell death. It also compares direct neutralization, inhibition of release or translocation, epigenetic/RNA-based regulation, and receptor-directed strategies. The evidence is predominantly derived from short-term rodent studies, with one large-animal antibody study and no therapeutic human trials. Accordingly, HMGB1 is best regarded as a biologically compelling but clinically unvalidated target whose therapeutic value will depend on redox- and phase-selective inhibition, kidney-directed delivery, and rigorous pharmacokinetic and safety evaluation.

Pharmaceuticals doi: 10.3390/ph19091357

Authors: Artyom Savelyev Dmitriy Khrustalev Irina Losseva Azamat Yedrissov Anastassiya Khrustaleva Vladimir Kazantsev Sofiya Shapovalenko Marlen Kiikbayev Polina Rusyaeva Kristina Perepelitsyna

Background: Topical semi-solid dosage forms are crucial for treating inflammatory skin disorders, wounds, burns, and superficial infections. Despite their widespread clinical use, comprehensive pharmaceutical market analyses of registered topical medicines remain limited in Central Asia. The aim of this study was to characterize the pharmaceutical market of topical semi-solid dosage forms registered in the Republic of Kazakhstan, including their dosage-form distribution, pharmacotherapeutic composition, manufacturer structure, and structural diversity of active pharmaceutical ingredients (APIs), with particular attention to heterocyclic compounds. Methods: A cross-sectional descriptive analysis was conducted using the State Register of Medicinal Products of the Republic of Kazakhstan. Registration records of topical semi-solid dosage forms were identified according to predefined inclusion criteria and classified by dosage form, pharmacotherapeutic category, manufacturer, country of origin, and API chemical structure. Descriptive statistical analysis, market concentration indices, and structural classification of heterocyclic compounds were applied. Therapeutic recommendations for dermatitis management from selected CIS countries were reviewed to provide a contextual interpretation of the observed market structure. Results: A total of 1046 registered topical semi-solid dosage forms met the inclusion criteria. Ointments (39.5%), gels (28.9%), and creams (25.4%) accounted for 93.8% of all registered products, indicating a highly concentrated dosage-form structure. Anti-inflammatory agents represented the largest pharmacotherapeutic group, whereas wound-healing and combination products accounted for smaller proportions of the market. Structural classification demonstrated broad representation of established heterocyclic chemical classes, while no registered medicinal products containing dihydropyrimidine-2(1H)-thione (DHPM-thione) derivatives were identified in the registry. Comparison with regional clinical recommendations indicated differences between recommended therapeutic approaches and the distribution of registered pharmacotherapeutic categories. Conclusions: This study provides the first comprehensive descriptive assessment of the registered market of topical semi-solid dosage forms in Kazakhstan. The results establish a reference dataset describing the current pharmaceutical landscape, including dosage forms, therapeutic categories, manufacturers, and API chemical classes. The absence of registered DHPM-thione-containing medicinal products indicates that this chemical class is currently not represented in the national medicines registry. Together with previously published preclinical evidence, this observation supports consideration of DHPM-thione derivatives as a subject for future pharmaceutical research, although the present study does not permit conclusions regarding their clinical effectiveness, market demand, or future commercialization.

Pharmaceuticals doi: 10.3390/ph19091356

Authors: Aleksandr Subbotin Holger A. Volk Sebastian Meller Gemma Mazzuoli-Weber Kristin Elfers

Background/Objectives: Antiseizure drugs (ASDs) are the primary therapeutic approach for epilepsy in small animals. Although ASDs are primarily used to modulate central neuronal excitability, they are commonly administered systemically, most often by the oral route, and may therefore influence neuronal populations outside the central nervous system. Nevertheless, their functional effects on peripheral neuronal populations, including enteric and dorsal root ganglion (DRG) neurons, remain incompletely characterized at a comparative pharmacological level. This study aimed to perform a comparative functional neuropharmacological profiling of commonly used ASDs in primary cultured myenteric and DRG neurons. Methods: Changes in neuronal activity were assessed in primary cultured guinea pig myenteric and DRG neurons using voltage-sensitive dye imaging with Di-8-ANEPPS following direct ASD application under standardized in vitro conditions. Results: ASDs exerted distinct drug- and neuron-type-specific effects on peripheral neuronal excitability. Topiramate induced the most pronounced reduction in neuronal excitability in myenteric neurons, whereas phenobarbital and levetiracetam produced only minor changes compared with buffer control. Potassium bromide induced mainly excitatory effects in both enteric and DRG neurons. Overall, most ASDs predominantly increased neuronal excitability in DRG neurons. Conclusions: These findings demonstrate distinct functional response profiles of ASDs in enteric and sensory neuronal populations. This comparative in vitro approach may provide a basis for future studies investigating peripheral neuronal drug effects and may help relate experimental pharmacological profiling to clinically relevant challenges associated with ASD treatment across different disorders.

Pharmaceuticals doi: 10.3390/ph19091355

Authors: Tamás Gáll Dávid Pethő Szilárd Póliska József Balla György Balla

Background/Objectives: Hypoxia is a major contributor to the development of retinal diseases by promoting metabolic adaptation and pathological angiogenesis through hypoxia-inducible factor (HIF) signaling. HIF-prolyl hydroxylase inhibitors (HIF-PHIs), which stabilize HIF under normoxic conditions, are widely used to treat anemia associated with chronic kidney disease. However, it remains unclear whether pharmacological HIF stabilization fully reproduces the cellular responses induced by physiological hypoxia. Methods: In this study, ARPE-19 cells were exposed to either physiological hypoxia or HIF-PHI treatment and analyzed using RNA sequencing, pathway enrichment, and pharmacological inhibition of PI3K/Akt, mTOR, and HIF-related signaling pathways. Results: Both treatments elicited a common transcriptional response marked by the induction of glycolytic and hypoxia-responsive genes together with increased vascular endothelial growth factor a VEGFA expression. Despite these similarities, only a small number of genes differed between the two conditions, suggesting that physiological hypoxia activates additional regulatory mechanisms beyond HIF stabilization alone. Rapamycin inhibited VEGFA production only under physiological hypoxia, further supporting mechanistic differences between the two models. Functionally, conditioned media from hypoxic ARPE-19 cells promoted endothelial tube formation, which was significantly reduced by pathway inhibition. Conclusions: Collectively, these findings show that HIF-PHIs reproduce many, but not all, aspects of the physiological hypoxic response in RPE cells.

Pharmaceuticals doi: 10.3390/ph19091354

Authors: Oscar Salvador Barrera-Vázquez Gil Alfonso Magos-Guerrero Juan Luis Escobar-Ramírez Rubén San Miguel-Chávez Maira Huerta-Reyes

Chiranthodendron pentadactylon Larreat (“flor de manita”) is traditionally used for gastrointestinal, cardiovascular, and neurological disorders. Objective: This study comprehensively integrated phytochemical characterization, in silico toxicological profiling, an exposure-informed Integrative Toxicity Prediction Model (ITPM), uncertainty analysis, and acute testing of a fresh flower aqueous extract (FFAE). Methods: Thirty-six phytochemicals were screened for hepatotoxicity, nephrotoxicity, Ames mutagenicity, carcinogenicity, hERG inhibition, reproductive-effects alerts, and acute oral toxicity. Five organ- or effect-specific endpoints were integrated into an exploratory Toxicological Alert Score (TAS), whereas reproductive alerts and predicted LD50 were reported separately. Seventeen compounds quantified in the FFAE were compared with a 20-compound literature-informed profile through 50,000 Monte Carlo iterations. Male CD-1 mice received a single FFAE dose of 300–5000 mg/kg and were observed for 14 days. Results: Positive predictions occurred for hERG inhibition in 33 compounds (91.7%), hepatotoxicity in 29 (80.6%), nephrotoxicity and carcinogenicity in 28 each (77.8%), and mutagenicity in 21 (58.3%). Six compounds had predicted LD50 values ≤ 1000 mg/kg. The proportional ITPM toxicity-evidence index was 6.91% for the experimental profile and 19.34% for the literature-informed profile. Monte Carlo means were 6.98% (95% uncertainty interval, 5.65–8.44%) and 19.32% (16.76–22.37%), respectively. No mortality, overt clinical signs, or treatment-related body weight differences occurred up to 5000 mg/kg. Conclusions: Compound-level predictions identified priorities for confirmatory testing, while the extract-specific model yielded a lower comparative toxicity-evidence proportion, and the animal study showed low overall acute toxicity under the evaluated conditions. These findings do not establish general or long-term safety, calibrated risk probabilities, or biological neutralization of toxicological liabilities.

Pharmaceuticals doi: 10.3390/ph19091353

Authors: Saied A. Aboushanab Denis A. Oberiukhtin Irina G. Danilova Pavel M. Vassiliev Ksenia V. Sokolova Kewei Chen Elena G. Kovaleva Alicia C. Mondragon Ekaterina I. Demicheva Jose M. Miranda

Background/Objectives: Type 2 diabetes mellitus (T2DM) is a multifactorial metabolic disorder characterized by chronic hyperglycemia, oxidative stress, protein glycation, low-grade inflammation, and dysregulation of signaling pathways, including those involving nuclear factor kappa B (NF-κB). Isoflavones from Fabaceae species exhibit diverse biological activities and may represent multifunctional candidates for addressing the interconnected mechanisms underlying T2DM. This study performed a consensus in silico pharmacodynamic screening of twelve representative Fabaceae isoflavones to prioritize compounds with predicted antiglycating, deglycating, antidiabetic, antioxidant, and NF-κB-related anti-inflammatory activities. Methods: Twelve representative Fabaceae isoflavones were evaluated using a multimethod computational workflow. Antiglycating and deglycating activities were predicted using IT Microcosm, antidiabetic and hypoglycemic activities were assessed using Prediction of Activity Spectra for Substances (PASS), antioxidant potential was evaluated using similarity-based prediction and quantum-chemical calculations, and NF-κB-related anti-inflammatory activity was investigated using similarity-based prediction combined with molecular docking. Molecular docking was applied specifically to the NF-κB-related anti-inflammatory analysis and did not contribute to the hypoglycemic/antidiabetic prediction. The individual predictions were integrated using a consensus scoring strategy to prioritize compounds across multiple pharmacodynamic domains. Results: The investigated isoflavones demonstrated moderate but broad predicted multifunctional activities. Malonylgenistin, biochanin A, and formononetin showed the most favorable predicted antiglycating profiles. Genistin, acetyldaidzin, acetylglycitin, and puerarin ranked highest in terms of predicted antidiabetic activity, whereas genistein exhibited the strongest predicted antioxidant profile. Daidzin and genistin received the highest integrated NF-κB-related computational rankings, although these predictions apply to intact glycosides rather than their expected in vivo hydrolysis products. Conclusions: This study presents a consensus computational framework for the comparative prioritization of Fabaceae isoflavones based on their predicted multifunctional pharmacodynamic profiles. The results provide a rational basis for selecting candidate compounds for future biochemical, cell-based, pharmacokinetic, and in vivo investigations, although it is emphasized that the present findings are based exclusively on computational predictions and require experimental validation.

Pharmaceuticals doi: 10.3390/ph19091352

Authors: Pasquale Degennaro Imane Ghafir El Idrissi Rosa Purgatorio Annalisa Fanizzi Mariagrazia Rullo Leonardo Pisani Eleonora Macchia Luisa Torsi Angela Stefanachi Francesco Leonetti

Proteolysis-Targeting Chimeras (PROTACs) are heterobifunctional molecules that induce the selective degradation of a protein of interest by recruiting an E3 ubiquitin ligase, thereby triggering ubiquitination and proteasomal clearance. As part of the broader targeted protein degradation (TPD) paradigm, PROTACs offer a powerful strategy to eliminate pathogenic proteins that are difficult to modulate with traditional occupancy-based inhibitors. However, their clinical translation is often limited by poor aqueous solubility, suboptimal cellular permeability, and off-target effects. Notably, some PROTACs retain potent biological activity despite limited membrane permeability, owing to their catalytic mechanism of action, which allows even a small number of molecules reaching the target site to drive substantial protein degradation and produce important pharmacological effects. Growing evidence supports the application of PROTAC-based approaches in neurodegenerative diseases, where the selective removal of toxic or misfolded proteins is particularly attractive. This review summarizes recent advances in chemical protein degradation strategies for neurodegenerative disorders and highlights potential future perspectives of multifunctional PROTACs for therapeutic development.

Pharmaceuticals doi: 10.3390/ph19091351

Authors: Yi-Xuan Li Xiao-Li Zhou Jing-Wen Xu Chen Zhao You-Yang Li Hai-You Zhao Liang-Peng Li Xin-Xin Tan Yu-Xiang Zhang Bei-Bei Yang Li Li Li-Hui Yin

Background/Objectives: Rotigotine is a conformationally flexible chiral dopamine agonist, but its solution-phase chiroptical behavior and conformational heterogeneity have not been systematically assessed. This study characterized its electronic circular dichroism (ECD), optical rotatory dispersion (ORD), and vibrational circular dichroism (VCD) responses and evaluated their consistency with its absolute configuration. Methods: ECD and ORD data were acquired for rotigotine ((S)-1) and its simplified analog (S)-2 in acetonitrile and methanol, and VCD spectra were recorded in appropriate deuterated solvents. Conformational searches, Boltzmann weighting, and density functional theory/time-dependent density functional theory calculations were performed. A principal-component-analysis/torsion (PCA-Tor) workflow selected a compact but structurally representative conformer ensemble for (S)-1. Results: In acetonitrile, (S)-1 exhibited a positive Cotton effect (CE) at 208.5 nm and a negative CE at 194.0 nm. The Boltzmann-weighted ECD spectrum reproduced the intense negative band below 200 nm and the positive band near 210 nm. The calculated VCD spectrum agreed with the signs and positions of the main experimental bands in the 1500–1100 cm−1 region. Experimental ORD curves of both compounds were negative in acetonitrile and methanol. The calculations reproduced the sign and overall trend, although their magnitudes were protocol- and population-sensitive. Ring puckering strongly affected the calculated ECD and ORD data, whereas side-chain flexibility expanded the accessible conformational space. Conclusions: The combined results support the known S configuration of rotigotine and demonstrate the value of representative conformational sampling for flexible chiral pharmaceuticals. This study also defines the practical utility and limitations of simplified molecular models in chiroptical analysis.

Pharmaceuticals doi: 10.3390/ph19091350

Authors: Jingyi Shen Xiaoya Liu Xuanyan Yan Tao Zhang Xianfang Zhang Huiquan Gu Weimin Chen Zhengwen Wang Qiang Liu

Hepatocellular carcinoma (HCC) remains difficult to control because recurrence, impaired hepatic reserve, and treatment resistance limit durable benefit. Natural products and traditional Chinese medicine (TCM) provide resources that range from drug-lead discovery to adjunctive multicomponent therapy. This review integrates pharmacological and clinical evidence for purified compounds, semisynthetic derivatives, extracts, formulas, and delivery systems. It focuses on metabolic reprogramming and redox homeostasis, stress responses and regulated cell death, tumor cell plasticity and vascular remodeling, and the immune microenvironment and host response. Recent studies have strengthened selected mechanistic claims through chemical probes, functional perturbation, and resistance models. Clinical research has concentrated on recurrence control after surgery or minimally invasive treatment and on combinations with transarterial chemoembolization, targeted agents, and immunotherapy. Randomized trials and prospective cohorts suggest potential benefit in specific settings, although product standardization, external validation, and long-term follow-up remain limited. Major translational barriers include uncertain active constituents, inadequate batch comparability, missing tumor-exposure data, and sparse herb–drug interaction studies. Future development should match target validation, pharmacokinetics, safety assessment, and clinical endpoints to each product class and clarify whether a candidate is best positioned as a drug lead, adjunctive therapy, or supportive intervention.

Pharmaceuticals doi: 10.3390/ph19091349

Authors: Zhihao Zeng Yanchang Liu Xiaoli Bi Wanchun Chen Yunhui Ouyang Jingnian Zhang Weitao Chen Guanlin Xiao

Background/Objective: Traditionally derived from the dried leaves of Microcos paniculata L., Microctis folium (M. folium) has been employed to manage various inflammatory and metabolic conditions. Despite its reported hypolipidemic potential, the active constituents contributing to this effect and their molecular mechanisms of action remain to be determined. Therefore, this study aimed to systematically identify the active hypolipidemic constituents of M. folium and elucidate their mechanisms of action through an integrated strategy combining chemical profiling, pharmacokinetic screening, network pharmacology, and experimental validation. Methods: UPLC fingerprint and UPLC-QQQ-MS/MS were employed to characterize and quantify the chemical constituents of M. folium. Pharmacokinetic analysis was conducted to identify systemically absorbed compounds. Network pharmacology was applied to predict potential targets and signaling pathways. The lipid-lowering effects were subsequently validated in vitro using an oleic acid/palmitic acid (OA/PA)-induced lipid accumulation model in HepG2 cells and in vivo in a Triton WR-1339-induced hyperlipidemia mouse model. Results: UPLC fingerprint analysis identified 15 common peaks among 21 batches of M. folium, with similarity values ranging from 0.885 to 0.990, indicating good chemical consistency. Eighteen representative compounds were quantified, among which flavone C-glycosides and phenolic acids were predominant. Pharmacokinetic results demonstrated that multiple M. folium’s constituents were absorbed into systemic circulation. Network pharmacology analysis identified 73 potential targets related to hyperlipidemia and highlighted the PI3K-Akt signaling pathway as a key regulatory pathway. In vitro experiments showed that M. folium’s compounds significantly reduced intracellular lipid accumulation and oxidative stress in OA/PA-induced HepG2 cells. Furthermore, in vivo studies demonstrated that vitexin, ferulic acid, isoferulic acid, and N-trans-feruloyltyramine significantly reduced serum and hepatic lipid levels, including TC, TG, and LDL-c, alleviated hepatic steatosis, and improved liver injury and oxidative stress markers in Triton WR-1339-induced hyperlipidemic mice. Conclusions: M. folium exerts significant lipid-lowering effects through multi-component and multi-target mechanisms involving regulation of lipid metabolism and oxidative stress, partly mediated by activation of the PI3K-Akt signaling pathway. These findings provide a scientific basis for the development of M. folium’s natural agents for the treatment of hyperlipidemia.

Pharmaceuticals doi: 10.3390/ph19091348

Authors: Qiuqi Zhuang Zhitao Yang Yan Liu

Objective: This study aimed to systematically evaluate the efficacy and safety of cadonilimab in the treatment of digestive system neoplasms. Methods: PubMed, Embase, Cochrane Library, Web of Science, Wiley Online Library, CNKI, Wanfang Data, CBM, and VIP databases were systematically searched to collect randomized controlled trials, non-randomized studies, and real-world studies investigating cadonilimab for the treatment of digestive system neoplasms. The initial search spanned from the inception of the databases to 4 February 2026, with an update search conducted in June 2026. Meta-analysis was performed using Stata 16.0 to systematically assess efficacy and safety outcomes. Results: A total of 10 studies, predominantly single-arm or retrospective, were included in this meta-analysis. Regarding the primary outcomes, because of the clinical heterogeneity across tumor types, pooled estimates should be interpreted as a summary across diverse diseases rather than as a single-tumor estimate. The overall pooled disease control rate (DCR) was 84% (95% CI = 76–91%), with tumor-specific DCRs of 87% in ESCC, 79% in HCC, 88% in G/GEJ adenocarcinoma, and 86% in PDAC. The pooled incidence of any-grade treatment-related adverse events (TRAEs) was 98% (95% CI = 95–100%), and the pooled incidence of any-grade immune-related adverse events (irAEs) was 32% (95% CI = 18–47%). For secondary outcomes: The partial response (PR) rate was 33% (95% CI = 21–46%), and the progressive disease (PD) rate was 11% (95% CI = 5–18%). The pooled incidence of grade ≥ 3 TRAEs was 52% (95% CI = 38–66%), and the pooled incidence of grade ≥ 3 irAEs was 11% (95% CI = 8–16%). However, incomplete reporting of low-frequency AEs across studies likely underestimates the pooled irAE rate. Conclusions: This study suggests that cadonilimab has the potential for antitumor activity in digestive system neoplasms. The observed safety profile was consistent with the known toxicities of immunotherapy, with notably high rates of TRAEs warranting vigilant monitoring. These findings should be considered hypothesis-generating and are insufficient to establish clinical benefit.

Pharmaceuticals doi: 10.3390/ph19091347

Authors: Zhihan Zhu Yu Liu Linglin Feng

Objective: Conventional progesterone (P4) formulations suffer from low bioavailability, severe local irritation, and poor patient adherence due to P4’s poor aqueous solubility. This work aimed to develop and screen a biodegradable PLGA/NMP (Poly(lactic-co-glycolic acid)/N-methyl-2-pyrrolidone) injectable in situ gel for sustained P4 delivery to overcome these clinical limitations. Significance: Commercial oral, vaginal, and oil-based intramuscular P4 preparations cannot maintain stable long-term drug exposure, and they cause injection-site pain/inflammation. The screened in situ depot system reduces administration frequency and local tissue irritation, supporting convenient luteal phase support and pregnancy maintenance. Methods: Nine formulations with variable P4 loading (10–50% w/w) and PLGA concentration (15–55% w/w) were fabricated. Formulations were screened via three core endpoints: injectability (injection force and discharge rate), in vitro sustained release in 10% Hydroxypropyl-β-cyclodextrin (HP-β-CD)-Phosphate-buffered saline (PBS) sink medium, and 7-day subcutaneous histocompatibility in rats. high-performance liquid chromatography (HPLC) was validated for progesterone quantification; Hematoxylin and eosin (H&E) staining assessed local inflammatory responses. Results: Formulations with progesterone ≤ 30% w/w and PLGA ≤ 35% w/w exhibited acceptable injectability (injection force < 50 N; discharge rate > 79%). Higher PLGA concentrations suppressed initial burst release (16.74% at 8 h for 35% PLGA vs. 29.8% for 20% PLGA). All formulations formed stable ellipsoidal subcutaneous depots and completed progesterone release within 4 days. Histopathology revealed only mild local inflammation (histological score = 1) without severe necrosis, superior to highly irritating oil injections in formulation control groups. Conclusions: The screened PLGA-based progesterone in situ gel resolves critical drawbacks of traditional progesterone dosage forms. This low-irritation, sustained-release injectable platform provides a scalable industrial formulation candidate for long-acting hormone therapy.

Pharmaceuticals doi: 10.3390/ph19091346

Authors: Mehmet Fatih Dasiran Hassen Daghmoura Ahmet Akbaş Yavuz Selim Angın Bakiye Akbaş Hatice Aygun Yiğit Uyanıkgil Oytun Erbas

Aim: Radiation-induced intestinal injury is one of the major dose-limiting complications of abdominal radiotherapy and is closely associated with oxidative stress, inflammation, and disruption of epithelial barrier integrity. The present study aimed to investigate the protective effects of verapamil on radiation-induced small-intestinal injury in rats by evaluating histopathological alterations together with tissue levels of TNF-α, 5-LOX, Nrf2, and ZO-1 as markers related to inflammatory, redox-associated, and tight-junction-associated responses. Materials and Methods: Thirty female Wistar albino rats were divided into three groups: Normal Control (n = 10), Radiation + Saline (RAD, n = 10), and Radiation + Verapamil (n = 10). Whole-abdomen irradiation was performed using a single 12 Gy dose delivered by a 6 MV linear accelerator. Verapamil (10 mg/kg/day, s.c.) treatment was started 5 days before irradiation and continued for 5 days afterward. Histopathological injury in jejunal tissues was evaluated using hematoxylin–eosin staining. Tissue levels of TNF-α, 5-LOX, Nrf2, and ZO-1 were measured using ELISA. Results: Whole-abdomen irradiation caused marked histopathological injury characterized by villus disruption, glandular degeneration, and mucosal architectural damage. Irradiation significantly increased small-intestinal TNF-α and 5-LOX levels while decreasing Nrf2 and ZO-1 levels compared with the Normal Control group (p < 0.05). Verapamil treatment significantly attenuated radiation-induced histopathological injury and reduced TNF-α and 5-LOX levels. TNF-α returned to values comparable with the Normal Control group, whereas 5-LOX remained significantly elevated. Verapamil also partially restored Nrf2 and ZO-1 levels compared with the RAD group (p < 0.05), although both remained significantly lower than control levels. Conclusions: Verapamil attenuated radiation-induced small-intestinal injury and was associated with favorable modulation of TNF-α, 5-LOX, Nrf2, and ZO-1 levels. The protective response was partial, with persistent differences in 5-LOX, Nrf2, and ZO-1 compared with controls. These biomarker changes should not be interpreted as direct evidence of functional pathway activation or preservation of intestinal barrier function. Further studies incorporating direct mechanistic and functional assessments are warranted before clinical translation.

Pharmaceuticals doi: 10.3390/ph19091345

Authors: Cody O’Rear Alejandro Chapa-Rodriguez Arminder Singh Safi Afzal Roshan Acharya Madalyn Wickham Usman Younus

Background: Direct oral anticoagulants (DOACs) are widely used among hospitalized patients, yet real-world inpatient bleeding data remains relatively sparse. Most evidence derives from randomized trials enrolling ambulatory, clinically stable patients, or those being treated for a single specific indication such as acute venous thromboembolism (VTE), which may not reflect the risk profile of acutely ill inpatients with multimorbidity, procedural exposures, and fluctuating renal function. This study describes the incidence and types of bleeding events in a real-world inpatient cohort receiving apixaban or rivaroxaban at a single tertiary care center. Methods: We conducted a single-center, retrospective cohort study of 867 hospital encounters involving apixaban (n = 722, 83.3%) or rivaroxaban (n = 145, 16.7%) between 2019 and 2021. The primary outcome was a non-adjudicated, non-standardized composite of any documented bleeding (gastrointestinal bleeding, hematuria, intracranial hemorrhage, vaginal bleeding, or epistaxis) during the index hospitalization; International Society on Thrombosis and Haemostasis (ISTH) major bleeding or clinically relevant non-major (CRNM) bleeding definitions were not applied. Secondary outcomes included individual bleeding subtypes, hemoglobin drop ≥2 g/dL, transfusion requirement, ICU/CCU admission, vasopressor use, and bleeding-related mortality. All comparisons are unadjusted and descriptive. Between-group comparisons used Fisher’s exact test and the Mann–Whitney U test. Results: Bleeding was documented in 13 encounters (1.5% overall; apixaban 1.39%, rivaroxaban 2.07%); given reliance on clinical documentation rather than systematic adjudication, true incidence may be higher. The composite outcome was driven predominantly by gastrointestinal bleeding (n = 10, 76.9% of events). No hemoglobin drop ≥2 g/dL, transfusion, ICU/CCU admission, vasopressor use, or bleeding-related mortality occurred in either group. No statistically significant between-agent difference was observed (p = 0.47); all comparisons are unadjusted and descriptive. Median length of stay (LOS) differed between drug groups (8.6 vs. 6.2 days; p = 0.006), an exploratory finding confounded by patient class imbalance. Conclusions: In this single-center, retrospective cohort, bleeding was documented in 1.5% of encounters; ascertainment limitations mean true incidence may be higher. Non-adjudicated, non-standardized outcome definitions and the small number of events limit comparability with the literature and preclude definitive inference. The absence of a statistically significant between-agent difference should not be interpreted as evidence of equivalence. These findings are hypothesis-generating and should not be used for comparative inference. Prospective, multicenter studies using ISTH-standardized outcomes, indication-level data, and formal adjudication are needed.

Pharmaceuticals doi: 10.3390/ph19091344

Authors: Danielle Alice Vieira da Silva Micaely Cristina dos Santos Tenório Fabiana Andréa Moura Nassib Bezerra Bueno Nathálya da Silva Severino Alexandra Rodrigues Bezerra Bianca Gomes de Souza Marilene Brandão Tenório Fragoso Tauane Alves Dutra Orlando Roberto Pimentel de Araújo Amylly Sanuelly da Paz Martins Marília Oliveira Fonseca Goulart Alane Cabral Menezes de Oliveira

Background/Objectives: This study aimed to evaluate the effects of oral administration of N-acetylcysteine (NAC) in pregnant women with preeclampsia (PE), using serum markers of redox imbalance and cytokine levels and perinatal outcomes. Methods: A double-blind, randomized, placebo-controlled pilot study was carried out from 2021 to 2023. The study was retrospectively registered with REBEC (Brazilian Clinical Trials Registry). After screening, women were randomized into 2 groups according to the type of supplementation: 600 mg of NAC (NAC group) or capsules containing 2% colloidal silicon dioxide and 98% microcrystalline cellulose (placebo group) until delivery. Blood samples were collected to quantify biomarkers of redox imbalance and cytokine levels. Questionnaires containing socioeconomic, clinical, obstetric, and anthropometric data were applied. Results: A total of 51 pregnant women with PE were included (26 in the placebo group and 25 in the NAC group). The use of NAC was associated with a higher gestational age at birth (GA) of the conceptus (placebo group: 37.39 weeks ± 1.96 SD vs. NAC group: 38.45 weeks ± 1.51 SD; p = 0.044). A moderate effect size was observed for this variable (Cohen’s d = 0.52). However, treatment allocation (NAC vs. control) was not independently associated with gestational age at birth after adjustment for covariates (F(1,28) = 0.144; p = 0.707; η2p = 0.005). The adjusted mean difference between groups was 0.92 weeks (95% CI: −0.16 to 2.00). No other significant effects on redox and inflammatory markers were identified using NAC. Conclusions: NAC supplementation did not impact the mode of delivery between the NAC and placebo groups. As a secondary exploratory finding, a possible trend toward prolonged gestational age was observed in the NAC group; however, this result should be interpreted with caution, given the exploratory nature of the analysis, the neutrality of the primary outcome, and the limited sample size.

Pharmaceuticals doi: 10.3390/ph19091343

Authors: Sıtkı Ün Ramazan Sabırlı İbrahim Türkçüer Gergana Lengerova Martina Bozhkova Steliyan Petrov Aylin Köseler

Background: Acute renal colic is a common urological emergency characterized by substantial interindividual variability in analgesic response. Pharmacogenetic variation in OPRM1, COMT, and ABCB1 may contribute to differences in opioid efficacy and pain control. This study primarily evaluated the associations of OPRM1 A118G (rs1799971), COMT Val158Met (rs4680), and ABCB1 C3435T (rs1045642) polymorphisms with opioid analgesic response in patients with acute renal colic. As a secondary exploratory analysis, genotype and allele frequencies were compared between patients and healthy controls. Methods: This prospective case–control study included 150 patients with acute renal colic and 100 healthy controls. Genotyping was performed using TaqMan SNP Genotyping Assays based on real-time polymerase chain reaction. Genotype frequencies were compared between groups using dominant and recessive genetic models, and Hardy–Weinberg equilibrium was assessed. In addition, genotype–phenotype associations were evaluated using pain severity, early analgesic response, initial opioid dose, rescue analgesic requirement, and multivariable logistic regression analyses. Results: In the secondary exploratory case–control analysis, no statistically significant differences in genotype or allele frequencies of OPRM1 rs1799971, COMT rs4680, or ABCB1 rs1045642 were observed between patients with acute renal colic and healthy controls. Within the patient cohort, however, genotype–phenotype analyses identified differences in early analgesic outcomes. Baseline-adjusted 30 min VAS differed according to OPRM1, COMT, and ABCB1 genotype, with the most pronounced difference observed for ABCB1 rs1045642. Patients with the ABCB1 TT genotype had higher adjusted 30 min VAS scores and showed a pattern of greater opioid requirement and more frequent rescue analgesia. In exploratory multivariable analysis, the ABCB1 TT genotype was associated with higher odds of inadequate early analgesic response (adjusted OR = 2.74, 95% CI 1.18–6.37; p = 0.019). Given the limited number of outcome events, this adjusted association should be considered preliminary and hypothesis-generating. Conclusions: No significant differences in the distributions of the polymorphisms investigated were observed between patients with acute renal colic and healthy controls. Within the patient group, ABCB1 genetic variation was associated with early opioid analgesic response, although this finding should be considered preliminary and requires confirmation in larger prospective pharmacogenetic studies before clinical implementation. Any potential future pharmacogenetic application should be considered as an adjunct to established first-line renal–colic management and specifically in patients for whom opioid therapy is clinically indicated.

Pharmaceuticals doi: 10.3390/ph19091342

Authors: Laura Schönfelder Shaoning An Peter Reusch Pedro M. Faustmann Timo Jendrik Faustmann Fatme S. Ismail

Background/Objectives: Glia-mediated inflammation contributes to a wide range of central nervous system (CNS) disorders, including epilepsy. Pregabalin (PGB) and cannabidiol (CBD) are CNS-acting drugs prescribed for various neuropsychiatric conditions, especially seizures. This study investigated the effects of PGB and CBD on glial properties in an astrocyte–microglia co-culture model of inflammation. Methods: Physiological (M5, containing 5–10% microglia) and pathological inflammatory astrocyte–microglia co-cultures (M30, containing 30–40% microglia) were collected from the postnatal brain hemispheres of Wistar rats (P0-P2) according to an established protocol and treated with different concentrations of PGB (3, 10, 30 and 60 µg/mL) for 24 h or CBD (50, 500 and 1000 ng/mL) for 1 h or 24 h. Metabolic activity was assessed by the MTT assay. Microglial phenotypes and astroglial connexin (Cx)43 expression were detected by immunocytochemistry. Results: In M5 co-cultures, short-term incubation (1 h) with high concentrations (1000 ng/mL) of CBD significantly reduced glial viability (p < 0.05), while no significant changes were observed in M30 co-cultures. After 24 h of incubation, M5 cultures exhibited a significant increase in metabolic activity at 50 ng/mL (p < 0.0001) and 500 ng/mL (p < 0.05), and a reduction at 1000 ng/mL (p < 0.05), suggesting impaired viability at high concentrations under physiological conditions. The distribution of microglial phenotypes in physiological M5 co-cultures incubated with CBD remained unchanged. In M30 co-cultures, CBD incubation for 1 and 24 h significantly reduced microglial activation and promoted a shift from reactive, phagocytic to homeostatic, ramified microglial phenotype (p < 0.05, p < 0.01, p < 0.0001). In contrast, PGB did not affect glial cell viability, microglial phenotypes or Cx43 expression in physiological and inflammatory co-cultures, indicating that the mechanisms of action of PGB probably do not include modulation of glial cells in vitro. Conclusions: The inhibition of microglial reactivity by CBD suggests potential positive effects on the neuroinflammatory component involved in the pathogenesis of CNS disorders such as epilepsy.

Pharmaceuticals doi: 10.3390/ph19091341

Authors: Xingli Zhang Huili Zhu Chengshu Yu Shuang Huang Dongdong Fang

Background/Objectives: Ovarian cancer is a prevalent gynecological malignancy with high mortality rates worldwide. Harmine hydrochloride (HMH), a tricyclic β-carboline alkaloid derived from the seeds of the medicinal plant Peganum harmala L., has demonstrated anti-tumor activity. This study investigated the effect of HMH on ovarian cancer and elucidated its underlying molecular mechanism. Methods: The anti-ovarian cancer effects of HMH were evaluated using the CCK-8 assay, cell cycle and apoptosis analyses, as well as a cell migration assay. The RNA-seq was utilized to explore the mechanisms of HMH. Western blot, qRT-PCR and IF were employed for further validation of the expression levels of relevant molecules. The Kaplan–Meier (KM) curve was conducted to evaluate the possible link between NFAT1 levels and the prognosis of ovarian cancer. The biological functions of NFAT1 were explored through siRNA knockdown experiments. Auto-dock molecular docking and DARTS were used to analyze the direct interaction between HMH and NFAT1. The in vivo efficacy of HMH was examined through bioluminescence imaging of intraperitoneal xenografts established with luciferase-expressing HEY cells. Results: HMH induced apoptosis, arrested the cell cycle, and suppressed migration of HEY and CAOV3 cells. NFAT1 was highly expressed in the nucleus of HEY and CAOV3 cells and promoted ovarian cancer progression. In addition, low NFAT1 expression levels had significantly improved overall survival and progression-free survival. Moreover, HMH reduced NFAT1 expression and reversed Ion-induced NFAT1 levels in ovarian cancer cell nuclei. The molecular docking and DARTS results suggested that HMH binds to NFAT1. Treatment with MG132 effectively blocked HMH-induced NFAT1 downregulation. Furthermore, HMH effectively blocked intraperitoneal xenograft development, and extended the life span of animals bearing HEY/Luc cells, when used alone or in combination with CDDP. Conclusions: The research indicated that HMH exerts anti-ovarian cancer effects, at least in part, by targeting NFAT1. Additionally, our research revealed that NFAT1 promotes ovarian cancer progression, and plays a significant role in HMH’s anti-cancer activity. These results may aid in the development of novel candidates for ovarian cancer and identify a new target for ovarian cancer treatment.

Pharmaceuticals doi: 10.3390/ph19091338

Authors: Zheming Xiong Yijie Su Luping Shi Shuyuan Ge Ge Wang Fangyu Li Xu Liu Jianguang Li Zejiang Ma

Background: Metabolic dysfunction-associated fatty liver disease (MAFLD) is a prevalent chronic liver disorder with limited pharmacological therapies, highlighting the need for novel multi-target agents from traditional medicinal plants. Artemisia rupestris L. has hepatoprotective effects, but its active constituents and mechanisms against MAFLD remain unexplored. Methods: The chemical profile of an ultrasonically-assisted 70% ethanol extract of Artemisia rupestris L. (ARE) was determined using UPLC-Q-TOF-MS. Network pharmacology, molecular docking, 100-ns molecular dynamics (MD) simulations, and in vitro experiments in oleic acid-induced HepG2 cells were integrated to identify active compounds, core targets, and pathways, and to validate lipid-lowering and hepatoprotective effects. Results: Seventy-eight compounds were identified, mainly flavonoids, phenolic acids, and terpenoids. Network analysis highlighted AKR1B10 and MAPK14 as core targets and isorupestonic acid and rupestonic acid as key components. MD simulations supported the stable binding of isorupestonic acid to AKR1B10 (RMSD ~1.6–3.2 Å), driven by van der Waals and electrostatic interactions. In vitro, ARE dose-dependently reduced intracellular triglycerides, total cholesterol, and LDL-C, decreased lipid droplets, and lowered AST/ALT levels, with a high-dose efficacy comparable to atorvastatin. Conclusions: Through in vitro experiments and network pharmacology analyses, this study preliminarily elucidated the beneficial effects of ARE on lipid deposition in hepatocytes and its potential mechanism, thereby providing an experimental basis and potential avenues for subsequent in vivo studies on pharmacodynamics, pharmacokinetics, and toxicology.

Pharmaceuticals doi: 10.3390/ph19091340

Authors: Musun Park Hyeun-Kyoo Shin Yujin Choi

Background/Objectives: Ephedrae Herba (EH) is widely used in traditional East Asian medicine, but safety concerns regarding its adverse events remain. This study aimed to investigate EH-associated adverse events using clinical pharmacovigilance data and to perform exploratory in silico analyses to propose potential molecular mechanisms underlying these adverse events. Methods: A disproportionality analysis was performed using individual case safety reports from the Korea Adverse Event Reporting System database. EH-containing products were compared with other herbal medicine products using reporting odds ratios (RORs), proportional reporting ratios, and information components. Network pharmacology identified adverse event-related genes and pathways, and protein–protein interaction networks were constructed. Molecular docking predicted direct adverse event-associated targets of ephedrine and compared mechanisms with control compounds (aconitine and spinosin). Results: Four adverse-event signals were detected in the primary analysis: sleep disorder, dry mouth, insomnia, and palpitations. Sensitivity analysis identified four signals, with three (dry mouth, insomnia, and palpitations) consistent across both analyses; constipation emerged only in the sensitivity analysis. Conclusions: Adverse event-associated network analysis predicted key pathways: Neuroactive ligand–receptor interaction, Pathways of neurodegeneration, and Dopaminergic synapse. Molecular docking predicted that ephedrine may act on downstream signaling mechanisms shared by neurotransmitter systems, including the dopaminergic system, distinct from control compounds.

Pharmaceuticals doi: 10.3390/ph19091339

Authors: Zike Xu Hongxia Luo Yimin Zhao Xuhui Wang Sha Chen

Background/Objectives: Cerebral ischemia–reperfusion injury (CIRI) poses therapeutic challenges because of oxidative stress, blood–brain barrier disruption, and neuronal apoptosis, limiting current treatments. Paeoniflorin (PF) from Paeonia lactiflora has neuroprotective potential, but its multi-target mechanisms remain unclear. This study investigated the role and mechanisms of PF in CIRI, focusing on neuron–endothelial crosstalk. Methods: Oxygen–glucose deprivation/reoxygenation (OGD/R) models were established using SH-SY5Y (human neuroblastoma) and HCMEC/D3 cells (human cerebral microvascular endothelial). Network pharmacology was used to predict potential PF targets and pathways. RNA sequencing, molecular docking, and molecular dynamics simulation were performed to screen and evaluate PF binding characteristics with key targets, and MTT, flow cytometry, Western blotting, and co-cultures were employed to detect paracrine interactions. Results: Network pharmacology and transcriptomics identified VEGF/PI3K-AKT/mTOR pathway enrichment. Molecular docking confirmed stable PF binding to VEGF-A (−8.4 kcal/mol), AKT (−5.5 kcal/mol), and mTOR (−9.6 kcal/mol). PF (10–80 μM) showed no cytotoxicity and reduced OGD/R injury in a concentration-dependent manner (maximal at 40 μM). PF activated VEGF/PI3K-AKT/mTOR signaling, reducing apoptosis by 57% (SH-SY5Y) and 33% (HCMEC/D3); PI3K inhibitor LY294002 abolished these effects. PF-treated HCMEC/D3-conditioned media enhanced OGD/R neuronal viability, verifying paracrine crosstalk. Conclusions: PF alleviated CIRI by directly protecting neurons and indirectly modulating neuron–endothelial crosstalk through VEGF/PI3K-AKT/mTOR activation, supporting its multi-target therapeutic potential.

Pharmaceuticals doi: 10.3390/ph19091337

Authors: Vladislav S. Sukhachev Sergey M. Ivanov Dmitry A. Filimonov Anastasia V. Rudik Vladimir V. Poroikov

Background/Objectives: Adverse drug reactions (ADRs) caused by drug–drug interactions (DDIs) represent an important problem in pharmacotherapy, especially in patients receiving multiple medications. Most computational approaches to DDI-associated ADR prediction formulate the task as a binary classification, but they do not explicitly consider the severity of adverse reactions. Our study aims to develop structure-based models that predict the severity of ADRs associated with specific drug pairs. Methods: Datasets were generated using DrugMAP as the source of drug pair–ADR associations with annotated severity categories, and TwoSides was used as an additional source to generate conditionally negative examples. The drug pairs were represented using PoSMNA descriptors, which encode pair-specific structural features derived from the molecular structures of both compounds. Predictive models were built using PASS DDI software. Model performance was evaluated using a modified cross-validation procedure that excluded compound-level overlap between the training and test sets, thereby reducing information leakage caused by the repeated occurrence of the same drugs in different pairs. Results: Models were developed for 14 clinically relevant ADR types, including cardiovascular, hepatotoxic, nephrotoxic, hemorrhagic, metabolic, and neurological effects. The unweighted class-level macro-average AUC values ranged from 0.830 for the Major category to 0.911 for the Minor category, while balanced accuracy ranged from 0.776 to 0.857. Predictive performance varied significantly between ADR types and severity categories. Higher accuracy was observed for some ADR types that were better captured by the structure-based descriptors used in this study, whereas complex multifactorial reactions, such as hepatotoxicity, were less accurately predicted. Case-based assessment using clinically documented drug combinations showed that the predicted severity profiles were generally consistent with the expected clinical risk patterns. Conclusions: The proposed approach demonstrates that the PoSMNA descriptors of drug pairs can be used for preliminary prediction of DDI-associated ADR severity. The developed models can help to filter out potentially dangerous drug combinations at an early stage and are implemented in the AdverDDIPred web-application.

Pharmaceuticals doi: 10.3390/ph19091336

Authors: Shini Zhang Xiao-Li Luo Hangxing Yang Ganghua Huang Jing Leng Tao Yang

Background: Alzheimer’s disease (AD) is a prevalent neurodegenerative disorder for which effective treatments remain limited. Liubao tea, a distinctive post-fermented dark tea, develops unique aged and areca nut aromas through prolonged storage. However, comprehensive studies on its anti-Alzheimer’s disease effects are lacking. This study aimed to identify the bioactive constituents of Liubao tea with varying storage durations and aroma profiles, and to further evaluate its neuroprotective potential against Alzheimer’s disease. Methods: Chromatography and spectrophotometry were employed to quantitatively determine the primary bioactive components in various Liubao tea samples. A D-galactose-induced Alzheimer’s disease zebrafish (Danio rerio) model was established to evaluate the neuroprotective effects of the aqueous extract of Liubao tea. Results: The contents of theabrownins, theaflavins, tea pigments, and catechins showed potential associations with the anti-AD capacity of Liubao tea. Variations in these bioactive components appeared to correlate with the neuroprotective activity. Conclusions: This study elucidates the anti-AD potential of aged Liubao tea and associates its characteristic constituents with neuroprotective effects. The findings provide essential theoretical support for developing Liubao tea as a promising natural product candidate worthy of further anti-AD research.

Pharmaceuticals doi: 10.3390/ph19091334

Authors: Yoon-Mi Lee Sumin Han Gyun Seok Park Judy Gopal Jae-Wook Oh

Background/Objectives: Colorectal cancer (CRC) remains a major cause of cancer-related mortality worldwide, largely owing to metastasis, recurrence, and resistance to conventional therapies. Natural-product-derived compounds represent an important source of anticancer drug candidates. This study investigated the anticancer activity of purified narciclasine, which was tentatively annotated as a major detected constituent of Lycoris sanguinea extract, and examined its association with HELLS-associated DNA damage responses in CRC cells. Methods: Human CRC cell lines HT29 and HCT116 were used to evaluate the cytotoxic and mechanistic effects of L. sanguinea extract and narciclasine. Cell viability, clonogenic growth, apoptosis, mitochondrial membrane potential, cell cycle distribution, migration, and invasion were assessed. Western blotting was performed to analyze apoptosis-, cell cycle-, epithelial–mesenchymal transition (EMT)-, and DNA damage-related proteins. UPLC-QTOF/MS analysis was used to identify major phytochemical constituents. HELLS knockdown and γ-H2AX immunofluorescence staining were conducted to examine the involvement of HELLS in narciclasine-associated DNA damage and apoptosis. Results: L. sanguinea extract reduced CRC cell viability, suppressed clonogenic growth, induced mitochondrial dysfunction and caspase-dependent apoptosis, and promoted G2/M cell cycle arrest with decreased cyclin B1 and increased p53/p21 expression. The extract also inhibited migration and invasion, accompanied by reduced β-catenin, N-cadherin, vimentin, slug, and snail expression and increased E-cadherin expression. UPLC-QTOF/MS tentatively annotated narciclasine as a major detected constituent. Purified narciclasine recapitulated several anticancer effects of the extract, accompanied by HELLS downregulation, increased γ-H2AX accumulation, reduced cyclin B1 expression, and enhanced PARP and caspase-3 cleavage. HELLS knockdown further sensitized CRC cells to narciclasine-associated γ-H2AX accumulation, G2/M arrest-associated signaling, and apoptosis. Conclusions: Narciclasine treatment was associated with HELLS downregulation, DNA damage-associated signaling, G2/M cell cycle arrest-associated changes, EMT suppression, and apoptosis in CRC cells. These findings suggest that narciclasine is a promising natural-product-derived lead compound for further preclinical evaluation in HELLS-associated CRC vulnerabilities.

Pharmaceuticals doi: 10.3390/ph19091335

Authors: Giorgio Cameli Alessia Piergentili Eleonora Spinozzi Alessia Tombesi Riccardo Petrelli Loredana Cappellacci Maria Beatrice Morelli

Background: Glioblastoma (GBM) is the most aggressive subtype of malignant glioma. Current therapeutic options remain limited, highlighting the need for the development of novel compounds capable of improving the efficacy of standard treatments. This study aimed to evaluate the biological activity of a series of pyrazolone-based hydrazone compounds (TPPs) in vitro GBM cell lines. Methods: The eight TPPs were synthesized by a nucleophilic addition reaction of different substituted hydrazines with 1-(5-hydroxy-3-methyl-1-phenyl-1H-pyrazol-4-yl)-2-phenylethan-1-one and tested on two human GBM cell lines, T98 and U87. The cytotoxic effects were evaluated via MTT assay. The most active compound was further investigated at IC50 and IC25 concentrations to evaluate mechanisms of cellular damage, including reactive oxygen species (ROS) production and mitochondrial membrane potential (ΔΨm) changes. Additional assays included colony formation, cell cycle analysis, and evaluation of DNA damage and apoptosis markers. Results: TPP25 exhibited the highest activity with IC50 values of 11.01 μM (95% CI: 10.42 to 11.64) and 13.12 μM (95% CI: 10.23 to 16.87) on T98 and U87 lines, respectively. Treatment induced early ROS production and mitochondrial depolarization, along with a significant reduction in colony formation. Cell cycle analysis revealed accumulation in the sub-G0 phase, consistent with increased cell death, supported by propidium iodide uptake. Furthermore, the results suggest the involvement of an apoptotic-like mechanism as supported by Annexin V positivity, γ-H2AX upregulation and transient caspase-3 activation. Conclusions: TPP25 demonstrates significant in vitro cytotoxicity, likely driven by a pro-apoptotic mechanism. This profile positions it as a potential lead compound for further preclinical evaluation, supporting its future transition into in vivo GBM models.

Pharmaceuticals doi: 10.3390/ph19091333

Authors: Adina Hamiti Yingying Geng Pengfei Huang Jinghui Xie Tingting Cui

Vitiligo is a chronic acquired depigmenting disorder characterized by progressive melanocyte loss. Its pathogenesis involves a multifactorial interplay among oxidative stress, impaired melanocyte regeneration, immune-mediated cytotoxicity, and neuroendocrine dysregulation. In recent years, traditional Chinese medicine (TCM)-derived active ingredients have attracted increasing research interest as potential interventions for vitiligo, although most isolated compounds remain at the preclinical stage. This review summarizes pharmacological and natural compounds applied in vitiligo treatment and highlights their regulatory mechanisms on core pathogenic modules. A comprehensive analysis of natural and semisynthetic active ingredients is provided, covering their preclinical and clinical evidence, mechanisms of action, and therapeutic relevance. Multiple ingredients, including baicalein, quercetin, curcumin, paeoniflorin, kaempferol, glycyrrhizin, and epigallocatechin-3-gallate (EGCG), have demonstrated antioxidant, anti-inflammatory, immunomodulatory, and neuroprotective effects that are involved in the pathogenesis of vitiligo. Some TCM-derived interventions, such as Ginkgo biloba L. extract and compound glycyrrhizin, have been evaluated clinically, whereas most isolated active compounds, including baicalein and quercetin, remain at the preclinical stage. Overall, TCM-derived active ingredients represent a promising therapeutic strategy for vitiligo. However, continued translational and clinical research is still required to optimize formulations, dosing regimens, and safety profiles, thereby facilitating their integration into routine clinical practice.

Pharmaceuticals doi: 10.3390/ph19091332

Authors: Jungha Min Sueun Shin Sohyeon Park Jeongha Yun Kaylnn Park Sandy Jeong Rhie

Background/Objectives: Real-world antidepressant use for depressive disorder varies across clinical settings. This study aimed to characterize prescribing pathways and evaluate the comparative safety of frequently prescribed antidepressants in a large multicenter cohort with newly diagnosed depressive disorder. Methods: We conducted a retrospective multicenter observational study using electronic health record data standardized to the OMOP Common Data Model (1999–2026) in adults initiating antidepressants. Prescribing patterns among 13,381 patients were visualized using sunburst plots and Sankey diagrams. Comparative safety, escitalopram vs. sertraline and trazodone, was evaluated across 18 hospitals (n = 44,542) for nine clinical safety outcomes using propensity score-matched Cox proportional hazards models and empirically calibrated random-effects meta-analysis. Results: Selective serotonin reuptake inhibitors, particularly escitalopram (21.4%, n = 2868), were the preferred antidepressant. Common adjunctive medications included trazodone (14.6%, n = 794) and quetiapine (14.0%, n = 761), though utilization varied across hospitals. No clear differences were detected for most safety outcomes between cohorts. However, empirically calibrated QTc prolongation risk was significantly increased with sertraline relative to escitalopram (HR 1.75, 95% CI 1.10–2.78), a signal requiring further confirmation. Conclusions: This study demonstrates substantial institutional variation exists in real-world antidepressant treatment pathways and combination strategies. While comparative safety profiles were broadly comparable across evaluated outcomes, the isolated QTc prolongation signal for sertraline warrants targeted external confirmation. Furthermore, trazodone and escitalopram cohorts should not be regarded as clinically interchangeable first-line options, supporting individualized, evidence-based antidepressant selection in routine clinical practice.

Pharmaceuticals doi: 10.3390/ph19091331

Authors: Goksu Kasarci-Kavsara Sinem Bireller Baris Ertugrul Bedia Cakmakoglu

Background/Objectives: Drug resistance is a major challenge in cancer therapy, and mitochondria contribute to this process by controlling both metabolic adaptability and cell survival signaling. Mitophagy, the selective lysosomal removal of dysfunctional mitochondria, has been implicated in therapy resistance, yet its role in sunitinib-resistant renal cancer remains poorly defined. Methods: In this study, acquired sunitinib resistance was established in ACHN renal cancer cells through eight months of stepwise dose escalation. Initial selection conditions were determined using CCK-8 viability and crystal violet colony assays in parental ACHN cells, whereas sustained proliferation under continuous sunitinib exposure was used as the operational criterion for the resistant phenotype. Resistant and parental sensitive cells were treated with 25 µM and 50 µM succinic acid, alone or in combination with sunitinib. Gene expression of BNIP3, NIX, FUNDC1, LC3, PINK1, Parkin, PGAM5, SRC, LONP1, and ATP5F1A was measured by RT-qPCR, and BNIP3 and NIX protein levels were assessed by ELISA. Results: Resistant cells showed significant upregulation of receptor-mediated mitophagy components BNIP3, NIX and FUNDC1 (p < 0.05), with no significant change in LC3, alongside suppression of PINK1, Parkin, and mitochondrial homeostasis-associated genes LONP1, PGAM5, and ATP5F1A (p < 0.05). Succinic acid predominantly reduced BNIP3 and NIX protein levels in both cell lines and suppressed BNIP3, NIX, and LC3 mRNA expression in resistant cells. In contrast, the sunitinib + 50 µM succinic acid combination selectively increased PARKIN, PGAM5, LONP1, and ATP5F1A expression in resistant cells (2.49- to 5.98-fold; p < 0.005), a pattern not observed in parental cells. Conclusions: These findings indicate that sunitinib resistance in ACHN cells is associated with upregulated transcription of receptor-mediated mitophagy components and downregulated transcription of PINK1/Parkin pathway genes, and that exogenous succinic acid selectively upregulates PARKIN and other mitochondrial homeostasis-related gene expression in resistant, but not parental, cells.

Pharmaceuticals doi: 10.3390/ph19091330

Authors: Ibrahim M. Aziz Rawan M. Alshalan Reem M. Aljowaie

Background/Objectives: The increasing global prevalence of chronic diseases has intensified the search for potential natural therapeutic agents. Fagonia arabica is a medicinal plant widely used in traditional medicine; however, comparative information on the phytochemical composition and biological activities of its leaves and seeds remains limited. To the best of our knowledge, this study represents the first comparative investigation of the phytochemical profiles and bioactivities of methanolic leaf (FALE) and seed (FASE) extracts of F. arabica collected from Riyadh, Saudi Arabia. Methods: Methanolic extracts were analyzed by gas chromatography–mass spectrometry (GC–MS), and their total phenolic content (TPC), total flavonoid content (TFC), antioxidant, antimicrobial, anticancer, and antidiabetic activities were evaluated using standard in vitro assays. Results: GC–MS analysis identified 60 and 68 compounds in FALE and FASE, respectively, with (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid, hexadecanoic acid, dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol, and (1R,3aR,5aR,5bR,7aR,9S,11aR,11bR,13aR,13bR)-3a,5a,5b,8,8,11a-Hexamethyl-1-(prop-1-en-2-yl)-1,2,3,4,5,6,7,7a,9,10,11,11b,12,13,13a,13b-hexadecahydrocyclopenta[a]chrysen-9-ol as major constituents. FASE exhibited higher TPC and TFC values (104.13 mg GAE/g and 62.16 mg QE/g, respectively) than FALE. Both extracts demonstrated antioxidant activity, with FASE showing greater potency against DPPH and ABTS radicals (IC50 = 63.3 and 42.4 μg/mL, respectively). FASE also displayed broad-spectrum antimicrobial activity, particularly against Staphylococcus aureus (MIC = 7.81 μg/mL). Furthermore, FASE exhibited stronger concentration-dependent cytotoxicity against MCF-7 and HepG2 cells, with IC50 values of 106.8 and 89.5 μg/mL, respectively, compared with 112.5 and 103.1 μg/mL for FALE. Apoptosis induction in MCF-7 and HepG2 cells was associated with upregulation of caspase-3, caspase-8, caspase-9, and Bax, alongside downregulation of Bcl-2 and Bcl-xL. In addition, FASE exhibited potent α-amylase and α-glucosidase inhibitory activities (IC50 = 61.62 and 67.38 μg/mL, respectively), comparable to acarbose. Conclusions: These findings highlight F. arabica seed extract as a promising source of multifunctional bioactive compounds with potential pharmaceutical application.

Pharmaceuticals doi: 10.3390/ph19091329

Authors: Ekaterina I. Alexeeva Natalia M. Kondrateva Tatyana M. Dvoryakovskaya Dmitry A. Kudlay Ivan A. Kriulin Maria S. Botova Irina T. Tsulukiya Elizaveta A. Krekhova Meiri S. Shingarova Maria Y. Kokina Anna N. Fetisova Tatiana Y. Kriulina Kseniya B. Isaeva Aleksandra M. Chomakhidze Christina V. Chibisova Mikhail M. Kostik

Introduction: Despite the growing spectrum of modern drugs for the treatment of juvenile idiopathic arthritis (JIA), patients with a refractory, progressive course of the disease remain a clinical challenge. B-cell-depleting therapy has demonstrated efficacy in adult rheumatoid arthritis and several pediatric rheumatic diseases, encouraging us to evaluate rituximab (RTX) for children with refractory JIA. Objective: This study aimed to assess the long-term efficacy and safety of rituximab therapy in children with refractory juvenile idiopathic arthritis. Methods: This retrospective, single-center, observational study included 106 patients with JIA who had failed previous therapy. The efficacy and safety of the therapy were evaluated at baseline and at 3, 6, 12, 24, 36, and 48 months post-initial rituximab infusion using the American College of Rheumatology Pediatric Criteria (ACRpedi 30/50/70/90), C. Wallace criteria for inactive disease/remission, and the JADAS71 activity index until patient completion or withdrawal from the study. The primary endpoint was achieving an ACRpedi50 response by 3 months, and the secondary endpoint was achieving inactive disease status according to C. Wallace criteria by 6 months of follow-up. Results: An ACRpedi50 response was achieved in 78/100 (78%) patients by month 3 of rituximab therapy, and inactive disease status was reached in 48/93 (52%) patients by month 6. More than half of the patients demonstrate inactive disease or remission at each observation point. During rituximab therapy, oral glucocorticoids were discontinued in 18/57 (32%) patients, and 28/57 (49%) patients were able to halve their corticosteroid dose (p = 0.03). Rituximab demonstrated an acceptable safety profile, with a total of 351 adverse events (non-serious AEs: 304/351, 87%) and a serious AE rate of 0.8 per 100 patient-years. Conclusions: Rituximab can reduce disease activity and lower the dose of oral glucocorticoids in patients with refractory JIA, serving as a viable therapeutic option with an acceptable safety profile.

Pharmaceuticals doi: 10.3390/ph19091328

Authors: Sara Vasović Lucija Vasović Nikola Martić Somyot Chirasatitsin Velibor Vasović Saša Vukmirović Nebojša Pavlović

Peptide therapeutics occupy a unique position between small organic compounds and large protein biomolecules, combining high specificity, strong pharmacological efficacy, and favourable safety profiles. Consequently, they have emerged as important therapeutic agents for a wide range of diseases, including metabolic and oncological disorders. However, oral administration of peptide drugs remains a major challenge due to extensive enzymatic degradation, low intestinal permeability, mucus entrapment, and presystemic metabolism within the gastrointestinal tract. This review provides a comprehensive overview of contemporary strategies for improving oral peptide delivery, with special emphasis on emerging pharmaceutical formulation technologies, bioinspired delivery systems and ingestible device-assisted approaches. A qualitative literature search was conducted using major scientific databases and included relevant publications available up to May 2026. The analysis identified the main barriers responsible for low oral bioavailability of peptide drugs, as well as promising approaches to overcoming these obstacles, including peptide modification, enzyme inhibition, permeation enhancement, mucolytic strategies, and advanced carrier systems. Special attention is given to multifunctional carrier systems, ingestible medical devices and bile acid-inspired technologies as emerging directions in oral peptide delivery. The convergence of pharmaceutical sciences, bioinspired formulation strategies and biomedical engineering is expected to accelerate the clinical translation of oral peptide formulations and enable their therapeutic potential to be fully exploited.

Pharmaceuticals doi: 10.3390/ph19091327

Authors: Mervenur Yavuz Firli R. P. Dewi İlknur Keskin Turan Demircan

Background: Drug repurposing represents an accelerated and cost-effective approach to discovering novel oncologic therapeutics. Here, we investigated the anticancer potential and underlying mechanisms of marbofloxacin (MBF), a veterinary fluoroquinolone (FQ), against breast cancer (BC) cells. Methods: The cellular impacts of MBF on cell viability, anchorage-dependent growth, tumorigenicity, migration, apoptosis, proliferation, senescence, and mitochondrial function were thoroughly characterized. To further elucidate its mechanistic activity, real-time qRT-PCR, untargeted LC-MS/MS-based metabolomics, network pharmacology, and molecular docking analysis were integrated. Results: MBF suppressed BC cell growth by inhibiting cellular proliferation and migration, disrupting mitochondrial membrane potential, and inducing ROS-mediated apoptosis and irreversible cellular senescence. These phenotypic impacts were accompanied by upregulation of tumor suppressors such as CDKN1A and PUMA and downregulation of oncogenes including MKI67, BIRC5, and BCL-2. Metabolomic analysis revealed broad suppression of biosynthesis-related metabolic pathways, characterized by the depletion of critical polyamines and nucleotide pathways. Network pharmacology and molecular docking analyses identified EGFR and HSP90AA1 as putative hub proteins potentially associated with the observed anticancer phenotype. Conclusions: These results provide initial evidence that MBF induces metabolic and molecular rewiring in BC, highlighting its promise as a repositionable therapeutic candidate.

Pharmaceuticals doi: 10.3390/ph19091326

Authors: Nawaf Alshammari Reyaz Hassan Mitesh Patel Mohd Adnan

Neurodegenerative diseases represent a major cause of disability and death, but early diagnosis, prognosis, and therapeutic monitoring are challenging due to biological heterogeneity and the absence of disease-specific biomarkers. Neurofilament light chain (NfL) is a highly sensitive fluid biomarker of neuroaxonal injury with well-established clinical utility in selected neurological disorders, especially in disease monitoring and prognostic evaluation. However, since NfL is not disease-specific, the interpretation has to be integrated with complementary molecular, imaging, and clinical biomarkers. Recent advances in genomics, epigenomics, transcriptomics, proteomics, metabolomics, microbiome profiling, and neuroimaging provide complementary information about the molecular and biological processes underlying neurodegeneration. Artificial intelligence (AI) and machine-learning approaches also allow the integration of these heterogeneous datasets for multimodal biomarker modeling. This review examines the biological and clinical relevance of NfL across major neurodegenerative diseases and critically discusses its combination with multi-omics, neuroimaging, and AI-based approaches. Special emphasis is placed on disease monitoring, prognosis, patient stratification, and therapeutic-response modeling, distinguishing established clinical applications from emerging research directions. The present review also addresses ongoing methodological challenges, including assay standardization, data harmonization, model interpretability, multicenter validation, and clinical translation. Finally, future potential is discussed for NfL-based multimodal biomarker frameworks in precision neurology.

Pharmaceuticals doi: 10.3390/ph19091325

Authors: Deniz Öğütmen Koç İbrahim Sarbay Melike Mercan Başpınar Lütfi Mangal Burcu Eda Arda Hande Sipahi

Background/Objectives: Orlistat is a gastrointestinal lipase inhibitor used for weight management. Although its established safety profile is largely characterised by fat malabsorption-related gastrointestinal events, post-marketing reports suggest a broader spectrum of adverse events. Given the increasing intersection between obesity pharmacotherapy and the management of non-alcoholic fatty liver disease/metabolic dysfunction-associated steatotic liver disease (NAFLD/MASLD), this study evaluated gastrointestinal, hepatobiliary, pancreatic, and anorectal adverse-event reporting signals associated with orlistat in the U.S. Food and Drug Administration (FDA) Adverse Event Reporting System (FAERS) and assessed concordance with regulatory prescribing information. Methods: Quarterly FAERS files from 2004 Q1 through 2026 Q1 were analysed; some retained reports had Initial FDA Received Date (FDA_DT) values dating back to 1999. Descriptive analyses included all orlistat-associated reports, whereas disproportionality analyses were restricted to primary-suspect reports. Sixty-five prespecified Medical Dictionary for Regulatory Activities (MedDRA) preferred terms (PTs) were screened. Reporting odds ratios (RORs), 95% confidence intervals (CIs), and proportional reporting ratios (PRRs) were calculated. A signal was defined as ROR > 2.0, lower bound of the 95% CI > 1.0, and n ≥ 3. Two-sided p values calculated using Fisher’s exact test on the corresponding 2 × 2 contingency tables were adjusted using the Benjamini–Hochberg false discovery rate procedure to address multiple testing. Signal-positive PTs were cross-referenced with prescribing information from three jurisdictions. Results: Overall, 30,454 deduplicated orlistat-associated reports were identified, including 16,684 primary-suspect reports. Thirty-eight of the 65 prespecified MedDRA PTs met the signal criteria, and all remained statistically significant after Benjamini–Hochberg false discovery rate correction (q < 0.05). The strongest signals were rectal discharge (ROR 1175.53; 95% CI 1093.44–1263.78) and steatorrhoea (ROR 1147.99; 95% CI 1058.12–1245.50); change in bowel habit also showed a high ROR (47.32; 95% CI 38.27–58.51). Unlabelled signal-positive PTs included constipation, faeces hard, cholecystitis, and irritable bowel syndrome; change in bowel habit showed jurisdiction-specific labelling discordance. Conclusions: Orlistat-associated FAERS signals included both expected fat-malabsorption-related events and additional gastrointestinal, hepatobiliary, pancreatic, and anorectal PTs, several of which were not explicitly represented in the evaluated regulatory labels. These findings reflect disproportionate reporting rather than incidence, absolute risk, or causality and require confirmation in independent clinical or epidemiological data sources. They do not, by themselves, support regulatory labelling changes.

Pharmaceuticals doi: 10.3390/ph19081324

Authors: Simona Ardelean Ioana Ciopănoiu Ioana Cuc-Hepcal Anda O. J. Samoila Corina Morodan Mihaela Borlea Silviu L. Constantinescu Oana Koppandi Sorina Ciurlea Carmen Tomoroga Adriana Ledeți Livia C. Borcan George A. Drăghici Paul Albu Cristina A. Dehelean

Most active pharmaceutical ingredients (APIs) reach their target by passive systemic distribution, so the dose required for efficacy at the lesion is set by what healthy tissue can tolerate; conventional dosage forms consequently produce pharmacokinetic profiles that oscillate between toxic peaks and sub-therapeutic troughs. Nanoparticulate carriers (liposomes, lipid nanoparticles, polymeric and inorganic systems, and biomimetic carriers) and hydrogels (natural, synthetic, supramolecular, and microgel-assembled) have emerged as the dominant strategies to address this, increasingly combined as hybrid nanoparticle–hydrogel constructs in which the gel provides locoregional retention and the nanoparticles provide cargo protection, intracellular delivery and stimuli responsiveness. Stimuli-responsive chemistries (pH, redox, enzyme, ROS, hypoxia, temperature, light, magnetic, ultrasound, glucose, and multi-stimuli logic) translate the molecular signatures of a disease into spatiotemporally controlled cargo release. This narrative review consolidates the state of the art (prioritizing 2022–2026) and departs from the conventional carrier-type survey in one respect: the literature is read along an explicit chain—disease cue, sensing chemistry, carrier architecture, release mechanism and kinetics, administration route, and clinical readiness—which exposes a variable that classification by carrier type conceals. Across all three material classes, what governs release behavior is not primarily the carrier chemistry but the identity of the released species (dissolved drug, drug from an embedded nanoparticle, an intact nanoparticle, and a matrix fragment) and the transport step that limits it. This is why power-law exponent analysis developed for dissolved drug fits particulate release poorly, why statistical goodness-of-fit cannot by itself establish a release mechanism, and why carrier class predicts clinical readiness less well than administration route and regulatory product type. Translational hurdles—CMC complexity, regulatory fragmentation, anti-PEG immunogenicity, and the structural mismatch between preclinical promise and clinical efficacy—are critically appraised in light of previously reported <1% delivery efficiency analysis. This review identifies converging strategies that could move stimuli-responsive controlled release from an aspirational outcome to a routine clinical reality.

Pharmaceuticals doi: 10.3390/ph19081323

Authors: Mujeeb Ul Naeem Syeda Farwa Naqvi Yousaf Khan Samina Aslam Syed Aminullah Azmatullah Khan Thoraya A. Farghaly Wajid Rehman

Background: Esophageal squamous cell carcinoma (ESCC) remains highly aggressive and continues to limit clinical treatment for substantial cancer-associated morbidity and mortality worldwide. Despite advances in therapeutic interventions the lack of effective molecularly targeted treatments continues to restrict clinical management beyond conventional chemotherapy and radiotherapy. Among these therapeutic targets the ribosomal S6 kinase 4 (RSK4) has gained considerable attention because of its critical involvement in ESCC progression, survival and proliferation, suggesting its potential as a potential target for anticancer drug development. Methods: A series of thiazole-clubbed pyrimidine linked hydrazone hybrids (1–14) were synthesized via 4-aminothiazole-5-carbohydrazide functionalized intermediates and fully characterized and evaluated for their inhibitory activity against RSK4. Results: Biological assessment demonstrated that the synthesized analogues exhibited remarkable potency, with IC50 values between 15.32 ± 1.35 and 54.61 ± 2.17 nM compared with the reference inhibitor BI-D1870 (IC50 = 33.16 ± 1.34 nM). Among the evaluated compounds, 2, 8, 9, 13 and 14 emerged as the most potent candidates and showed pronounced activity towards RSK4. For further insights into the molecular basis of their activity the lead candidates were subjected to computational investigations, such as molecular docking, molecular dynamics simulations, in silico ADMET and ProTox-3.0 characterization. The computational analyses provided structural and pharmacological insights into experimentally observed RSK4 inhibitory activity, like predicted interactions, stability dynamically and preliminary ADMET/toxicity characteristics. This study supports the need for further optimization and experimental validation of the identified RSK4 active lead candidates. Conclusions: These findings highlight the thiazole-clubbed pyrimidine-linked hydrazone scaffold as a potential chemotype for promising scaffolds targeting RSK4 lead discovery, and the identified candidates warrant further investigation into ESCC cellular models to establish anticancer efficacy and pathway-level activity.

Pharmaceuticals doi: 10.3390/ph19081322

Authors: Alaa M. Elsayad Khaled A. Elsayad

Background/Objectives: Human carbonic anhydrase (CA) isoforms are clinically relevant zinc metalloenzymes, but their highly conserved catalytic Zn2+ site makes isoform-selective inhibitor design challenging. This study developed an explainable multi-isoform QSAR workflow for prioritizing selective inhibitors of human CA I, CA II, CA IX, and CA XII, integrating PubChem experimental concordance and applicability-domain (AD) analysis to distinguish model-supported candidates from exploratory extrapolative hypotheses. Methods: Curated ChEMBL Ki datasets were standardized to pKi and modeled in KNIME using 4185 two-dimensional descriptors, Random Forest-based feature selection, and H2O.ai AutoML stacked ensembles. A matched 3200-compound four-isoform matrix supported direct selectivity profiling. Potent ChEMBL inhibitors (Ki < 10 nM) seeded 90% PubChem similarity expansion; analogues were scored across all four isoform-specific models, while PubChem records were filtered to retain only human isoform-specific Ki evidence. AD was calibrated using Morgan fingerprint similarity, descriptor-space distance, and leverage analysis. Candidates were prioritized by predicted potency, selectivity, SAR plausibility, SwissADME profile, structural alerts, and AD membership. Results: Models achieved held-out R2 values of 0.727, 0.719, 0.652, and 0.607 for CA I, CA II, CA IX, and CA XII, respectively. PubChem concordance identified 1098 prioritized compounds with assay records, including 847 with direct Ki evidence and 428 with complete four-isoform coverage; same-target agreement showed Pearson r > 0.86 and linear-fit R2 > 0.74. Test-set error rose from very-high-AD to low/out-of-domain classes. Conclusions: The workflow integrates QSAR prediction, PubChem concordance, explainable SAR, ADME triage, and AD-based reliability assessment. CA II and CA XII candidates showed the strongest support, CA IX showed intermediate support, and CA I candidates require cautious interpretation as low-domain exploratory hypotheses.

Pharmaceuticals doi: 10.3390/ph19081321

Authors: Sandro La Vignera Rosita Condorelli

Background/Objectives: Male hypogonadism affects 35–50% of men with type 2 diabetes mellitus (T2DM) and obesity. The relationship between testosterone deficiency and response to incretin-based therapies (GLP-1 and GIP receptor agonists) remains incompletely understood. This systematic literature review, conducted following PRISMA 2020 guidelines, examines whether male hypogonadism represents a risk factor for poor response to incretin therapy or, conversely, whether these agents offer therapeutic benefits for this population. Methods: A comprehensive systematic literature search was conducted on 31 December 2024, across PubMed/MEDLINE, Scopus, and Web of Science using three search domains: (1) hypogonadism and incretin therapy response; (2) testosterone deficiency and GLP-1/GIP receptor agonists; (3) incretin response and testosterone. Eligibility criteria included human studies (randomized controlled trials [RCTs], observational studies, cohort studies, cross-sectional studies, systematic reviews, and meta-analyses) in adult men reporting testosterone levels and/or incretin response. Animal studies, pediatric populations, case reports with n < 5, editorials, and letters without data were excluded. Study selection followed PRISMA 2020 guidelines with independent dual screening. Risk of bias was assessed using the Cochrane Risk-of-Bias 2.0 tool (ROB2) for RCTs, the Newcastle–Ottawa Scale (NOS) for observational studies, and AMSTAR-2 for systematic reviews and meta-analyses. Due to substantial heterogeneity in study designs, populations, interventions, and outcome measures, a meta-analysis was not feasible; therefore, a narrative synthesis was performed. Results: From 326 records identified, 29 studies were included after deduplication and screening (primary studies: 14; systematic reviews/meta-analyses: five; narrative reviews/expert opinion: 10). Risk-of-bias assessment revealed moderate-to-high overall risk: RCTs had small sample sizes (n = 12–42) and short follow-up (12–24 weeks), raising concerns about statistical power; observational studies were of fair-to-good quality (NOS 4–7 stars) but subject to confounding; and systematic reviews were of low-to-moderate confidence (AMSTAR-2). Primary evidence from RCTs and observational studies demonstrates that GLP-1 receptor agonists (GLP-1RAs)—particularly semaglutide and liraglutide—and the dual GIP/GLP-1 receptor agonist tirzepatide significantly increase total testosterone levels in men with obesity-related functional hypogonadism (mean increase 2.5–5.2 nmol/L). This effect is largely mediated by weight loss and improvement of insulin resistance rather than direct androgenic action. Evidence regarding whether baseline hypogonadism impairs glycemic or weight-loss response to incretin therapy is limited and indirect; no adequately powered comparative trials stratified by baseline testosterone status were identified. Conclusions: Incretin-based therapies, particularly GLP-1 receptor agonists and the dual GIP/GLP-1 receptor agonist tirzepatide, appear to improve testosterone levels in men with obesity-related functional hypogonadism, an effect that is largely mediated by weight loss and improvement of insulin resistance rather than a direct androgenic action. However, direct comparative evidence between hypogonadal and eugonadal men regarding glycemic or weight-loss response to incretin therapy remains insufficient to draw firm conclusions. The hypothesis that male hypogonadism does not impair incretin therapy response is biologically plausible but is currently supported only by indirect evidence. Prospective, adequately powered trials stratified by baseline testosterone status are needed to resolve this question.

Pharmaceuticals doi: 10.3390/ph19081320

Authors: Anh Tuan Do Quoc Long Pham Huong Thi Thu Phung Minh Quan Pham

Background/Objectives: Cyclin-dependent kinase 2 (CDK2) is a key regulator of cell-cycle progression and a potential anticancer target. This study aimed to identify alkaloid-derived CDK2 ligands using an integrated computational workflow and to obtain preliminary evidence of their effects on cancer-cell viability. Methods: Molecular docking with mVina and fast pulling of ligand (FPL) simulations were benchmarked using 20 experimentally characterized CDK2 inhibitors. A library of 2692 PubChem-derived alkaloids was screened, followed by ADMET evaluation, 100 ns molecular dynamics simulations, and FPL-based relative-affinity re-ranking. The three prioritized compounds were evaluated in HepG2 and HGC-27 cells using an MTT assay after 48 h of exposure. Results: Docking and FPL showed correlations with experimental affinity data of RDock = 0.549 ± 0.180 and RW = −0.676 ± 0.119, respectively. CID 636885, CID 46184320, and CID 101691758 were prioritized for detailed evaluation. All three compounds reduced cell viability, with lower IC50 values observed in HepG2 cells than in HGC-27 cells. CID 101691758 exhibited the highest growth-inhibitory activity among the tested compounds, with IC50 values of 15.37 ± 0.46 µg mL−1 in HepG2 cells and 52.64 ± 1.33 µg mL−1 in HGC-27 cells. Conclusions: The workflow identified three preliminary alkaloid hits, with CID 101691758 showing the most favorable combined computational and cell-viability profile. However, the MTT assay does not establish direct CDK2 inhibition or kinase selectivity. Biochemical CDK2 inhibition, target-engagement, and kinase-panel studies are therefore required.

Pharmaceuticals doi: 10.3390/ph19081319

Authors: Ziying Xu Wei Xia Haiqiang Wu Haiping Zhang

Background/Objectives: Blood–brain barrier (BBB) permeability is a major practical obstacle in central nervous system (CNS) drug discovery, because only a small fraction of drug-like molecules achieve sufficient brain exposure. Methods: We present DeepBBB, a graph-based, data-composition-aware screening workflow for predicting BBB permeability and for constructing BBB-focused screening libraries from commercial chemical space. Rather than introducing a new graph-learning architecture, the workflow combines standard graph convolutional and graph-transformer models with deliberate control of training-set composition, commercial-library filtering, chemical-space profiling, and prospective experimental evaluation. Three model variants were trained on the Blood–Brain Barrier Database (B3DB): a baseline classifier/regressor pair (DeepBBB_V1_BC/RG), a variant trained with a more strongly negative-enriched configuration (DeepBBB_V2_BC), and a graph-transformer counterpart (DeepBBB_trans_BC/RG). Because the sample-level split assignments and per-compound predictions from the original runs were not recoverable, the archived summary metrics are reported descriptively in the main text and are not used to support calibration, scaffold-level validity, or generalization. Applying the workflow to the ChemDiv collection (~1.5 million compounds) and the Enamine REAL lead-like space (~1.7 billion compounds) produced three progressively more stringently filtered BBB-focused libraries (21,991; 4,808,885; and 151,790 compounds). Results: Analysis of available processed data indicated that the predicted BBB-permeable set occupies a compact, BBB-compatible property region. Physicochemical, fragment, and scaffold summaries were interpreted descriptively at the constructed-library level. In a first prospective campaign, one of 12 tested candidate compounds was PAMPA-BBB-positive (all-tested molecular-level positive fraction 8.3%; exact 95% CI 0.2–38.5%). In a second campaign, five of 35 tested candidate compounds were PAMPA-BBB-positive (14.3%; exact 95% CI 4.8–30.3%); 13 compounds were not quantifiable and were not treated as ordinary CNS-negative measurements, and the two campaigns differed in compound source, selection strategy, and assay setting, so the numerical difference is reported descriptively rather than causally. Conclusions: Together, these results support the feasibility of BBB-focused computational filtering and a PAMPA-BBB evaluation workflow for CNS-oriented discovery.

Pharmaceuticals doi: 10.3390/ph19081318

Authors: Normand García-Hernández Israel Ramírez-Sánchez Rosa María Ordoñez-Razo

The study of plant-derived secondary metabolites has advanced the field of pharmacochemistry [...]

Pharmaceuticals doi: 10.3390/ph19081316

Authors: Georgios D. Panos

Background: Faricimab 6 mg and aflibercept 8 mg permit extended treatment intervals in neovascular age-related macular degeneration (nAMD), but their pivotal programmes used different loading and maintenance schedules. This study compared treatment burden under the same loading and treat-and-extend protocol and included an exploratory longitudinal analysis of published aggregate visual and anatomical outcomes. Methods: Treatment-specific Dirichlet distributions represented persistent Q8W, Q12W and Q16W capability. Both treatments received injections at weeks 0, 4, 8 and 16, followed by identical four-week extensions from Q8W to Q16W. The primary analysis comprised 30,000 evidence draws, 100,000 paired virtual eyes, a 1.5-million-pair scenario grid and a nested five-million-pair probabilistic analysis. Exploratory multilevel meta-regressions estimated BCVA and change in retinal thickness through week 52; IRF, SRF and complete retinal dryness were analysed separately. Results: Mean injections with faricimab and aflibercept 8 mg were 7.255 and 7.212 at week 52 and 11.522 and 11.144 at week 104. The nested 104-week difference was −0.393 injection (95% uncertainty interval −0.682 to −0.107), but broad source-weight uncertainty included no difference and credible source analyses changed the direction of the contrast. Exploratory week-52 BCVA gains were 5.65 and 6.29 letters, respectively; the difference was 0.64 letter (95% uncertainty interval −1.41 to 2.70) and should be interpreted in the context of three aflibercept 8 mg study families. The expanded retinal-thickness sensitivity model estimated changes of −171.8 and −147.2 micrometres, while the between-treatment direction varied in the three-dose-only analysis. Complete retinal dryness was projected in 57.0% and 69.0% of eyes, respectively, based on three study families per treatment. Conclusions: Use of the same protocol predicted essentially equal one-year burden and a small two-year fixed-weight difference that varied across source choices. The exploratory longitudinal projections, informed by three independent aflibercept 8 mg study families at one year, provide supportive context for the visual and anatomical findings but should be interpreted as hypothesis-generating rather than confirmatory.

Pharmaceuticals doi: 10.3390/ph19081317

Authors: Ji Hye Hwang Chul Jung

Background/Objectives: Skin aging involves interconnected inflammatory, oxidative, hormonal, extracellular matrix (ECM), and cellular senescence-related mechanisms, supporting the need for multitarget approaches. This study aimed to evaluate the effects of a multicomponent natural product-based injectable formulation developed in Korean medicine practice, Dong-An Pharmacopuncture (DAP), on skin aging-related pathways. Methods: A network pharmacology approach was used to identify the active compounds, predicted molecular targets, and signaling pathways associated with DAP. Sixty-two active compounds from 11 constituent materials were screened, and 70 final targets were identified using STITCH-based prediction, intersection with GeneCards-derived skin aging-related targets, and quality filtering. Results: Herb-compound-target network analysis yielded 225 compound–target interactions across 292 edges. Protein-protein interaction analysis identified a highly connected network with 1,334 edges, and hub analysis converged on 10 core targets: TNF, IL6, ESR1, TP53, AKT1, PPARG, EGFR, PTGS2, CASP3, and PPARA. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses identified four major mechanistic axes: inflammatory and oxidative stress regulation, hormonal skin homeostasis, tissue repair and ECM remodeling, and cellular senescence-related regulation. Because DAP is administered by injection, a Good Laboratory Practice-compliant single-dose subcutaneous toxicity study was additionally conducted in Sprague–Dawley rats, which showed no mortality, abnormal clinical signs, or histopathological findings attributable to DAP at 1.0 mL/head. Conclusions: The findings in this study provide a systems-level framework for the predicted multitarget mechanisms of DAP in skin aging-related pathways and support the need for further experimental validation of its predicted mechanisms and repeated-dose safety.

Pharmaceuticals doi: 10.3390/ph19081315

Authors: Zineb Ariba Houda Zentar Mohammed Zerrouk Safaa Hidaoui Mohammed Er-rajy Meryem Hrimla Mustapha Ait El Had Rachid Chahboun Juan Sainz Rachid Ouarsal Mohammed Lachkar Fernando J. Reyes-Zurita

Background/Objectives: Metal-based coordination compounds are attracting attention in medicinal inorganic chemistry because changes in the metal centre can influence their physicochemical properties and biological responses. This study aimed to evaluate two isostructural ammonium-M(II) hydrogenophosphite hydrates, (NH4)2[Mg(H2O)6]3(HPO3)4 (complex 1) and (NH4)2[Co(H2O)6]3(HPO3)4 (complex 2), as bioactive systems with potential anticancer and anti-inflammatory properties. Methods: The complexes were synthesized and characterized by X-ray powder diffraction (XRPD), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), density functional theory (DFT) calculations, and Hirshfeld surface analysis. Their cytotoxic activity was evaluated against B16-F10, HT29, HepG2, and HL-60 tumour cell lines. For complex 2, apoptosis, cell-cycle distribution, and mitochondrial membrane potential were analysed by flow cytometry. Nitric oxide production was measured in LPS-stimulated RAW 264.7 macrophages. Results: Complex 2 showed the highest cytotoxic activity, particularly against HL-60 leukaemia cells (IC50 = 36.98 μg/mL), whereas complex 1 displayed weaker activity. Complex 2 increased apoptotic cell populations, altered cell-cycle distribution, and induced mitochondrial membrane depolarization in HL-60 cells. Both complexes reduced nitric oxide production, with complex 2 showing the strongest effect (IC50 NO = 22.46 μg/mL), exceeding that of diclofenac under the experimental conditions, while complex 1 showed comparable activity to the reference drug. DFT descriptors indicated higher electronic reactivity for complex 2. Conclusions: Replacement of Mg(II) by Co(II) enhances the biological activity of this hydrogenophosphite framework. Complex 2 combines cytotoxic activity against HL-60 cells with apoptosis-associated mitochondrial dysfunction and NO-inhibitory activity in activated macrophages, supporting hydrogenophosphite-based metal complexes as bioactive coordination compounds with potential as multifunctional agents.

Pharmaceuticals doi: 10.3390/ph19081314

Authors: Elena Coyago-Cruz Gabriela Méndez Johana Zúñiga-Miranda Alejandro Porras Carlos Barba-Ostria Ivanna Sarabia Linda Guamán Celina Coello Jorge Heredia-Moya Blanca Naranjo María Segovia-Salcedo

This exploratory study compared the flowers, leaves, and roots of Hypochaeris sessiliflora to evaluate physicochemical properties, bioactive compounds, in vitro chemical antioxidant activity, antimicrobial activity, and locomotion-disrupting effects on Caenorhabditis elegans. Bioactive compounds were determined by liquid chromatography; in vitro chemical antioxidant activity by microplate spectrophotometry; antimicrobial activity against a range of ATCC and multidrug-resistant microorganisms; and locomotion-disrupting effects on Caenorhabditis elegans. The results revealed significant differences between plant organs. The leaves contained the highest concentrations of calcium (1163.1 mg/100 g DW), potassium (3256.6 mg/100 g DW), vitamin C (39.2 mg/100 g DW), organic acids (1213.5 mg/100 g DW), and total quantified chlorophylls (89.0 mg/100 g DW). The flowers had the highest levels of total quantified carotenoids (152.1 mg/100 g DM) and total quantified phenolic compounds (12,341.1 mg/100 g DM). They exhibited the highest in vitro chemical antioxidant activity, as measured by the ABTS assay (6.1 mmol TE/100 g DW). The extracts exhibited antimicrobial activity that depended on the plant organ and the microorganism tested, with the root extract being the most active against Staphylococcus epidermidis (MIC = 8.2 mg/mL). No activity was detected against Candida species or against multidrug-resistant bacteria under the experimental conditions used. Furthermore, the extracts under the experimental conditions used did not significantly alter C. elegans’ locomotion. Overall, this study provides baseline evidence of the organ-specific chemical composition and biological properties of H. sessiliflora, supporting further investigation of the compounds responsible for the observed in vitro chemical antioxidant and selective and antimicrobial activities.

Pharmaceuticals doi: 10.3390/ph19081313

Authors: Caterina Nela Dumitru Teodora Marcu Alina Oana Dumitru Simona Steliana Tudor Ionela Daniela Ferțu Alina-Mihaela Elisei Larisa Goroftei

Background: Berberine, an isoquinoline alkaloid present in Berberis spp., Coptis chinensis and Hydrastis canadensis, is among the most widely consumed metabolic-health supplements, popularized as “nature’s Ozempic”. Concurrent, often undisclosed use with prescription drugs is common in older adults, yet berberine is far from inert. Objective: To synthesize the evidence on berberine as a perpetrator of supplement–drug interactions, propose a four-axis mechanistic taxonomy, with product quality treated separately as a modifier of exposure rather than as a mechanism, and derive a clinically actionable risk-stratification framework. Methods: Structured narrative review, prepared per the SANRA quality criteria; PubMed/MEDLINE, Scopus, Web of Science and Embase were searched up to May 2026. Results: Despite very low systemic exposure (oral bioavailability 0.68% in rats; low ng/mL plasma concentrations in humans), high luminal, enterocytic and hepatic concentrations generate interaction liability, documented in humans for a few pairs and mechanistic for most, along four mechanistic axes: inhibition, and transcriptional induction, of CYP3A4, with CYP2D6/CYP2C9 inhibition that is quasi-irreversible through a metabolite-intermediate complex; transporter modulation (P-glycoprotein, OCT1/OCT2, and MATE1); pharmacodynamic additivity (hypoglycemia, hypotension, and QT prolongation); and microbiome- and gut-barrier-mediated effects, the last of these being a candidate axis rather than a demonstrated one. Product-quality variability is treated separately, as a modifier of exposure. The clinical anchor is increased cyclosporine exposure in renal-transplant recipients (AUC +34.5%; trough 29.3% above control). These elements are integrated into a three-tier risk-stratification framework that combines perpetrator potency, victim-drug vulnerability, and patient vulnerability, with each tier being linked to a defined pharmacy action. Conclusions: In patients on multiple medications, and particularly when berberine is co-administered with drugs of narrow therapeutic index, it should be managed as an active pharmacological perpetrator rather than as an inert supplement. Unstandardized product quality and an unsettled European regulatory framework, under which national limits differ by more than an order of magnitude, further widen the uncertainty around the dose actually delivered. Berberine use should therefore be elicited routinely at medication reconciliation and stratified by mechanism, by victim-drug vulnerability, and by patient risk, with particular attention to metabolic self-medication in the GLP-1 era.

Pharmaceuticals doi: 10.3390/ph19081312

Authors: Yukiko Kurihara Hiroki Kurihara

G protein-coupled receptors (GPCRs) are dynamic allosteric proteins whose signaling properties are governed by reciprocal communication between extracellular ligand-binding sites and intracellular transducer interfaces. Although classical pharmacological models established the concept that ligand binding and G protein coupling are thermodynamically linked, recent structural, biophysical, and computational studies have revealed a far more complex picture in which GPCRs exist as ensembles of interconverting conformational states. Accumulating evidence indicates that G proteins function not only as downstream signaling effectors but also as endogenous allosteric modulators. By reshaping receptor conformational landscapes, G protein coupling can influence the structure and dynamics of orthosteric ligand-binding pockets, thereby regulating ligand affinity, binding kinetics, and receptor selectivity. These findings support a bidirectional model of GPCR signaling in which information is transmitted not only from ligand-binding sites to intracellular signaling partners but also in the reverse direction through receptor-wide allosteric networks. Disease-associated mutations of endothelin A receptor (ETAR) provide in vivo evidence that structural perturbations located far from orthosteric ligand-binding sites can alter ligand recognition through long-range allosteric communication. In addition, emerging studies of positive allosteric modulators demonstrate the therapeutic potential of selectively stabilizing ligand–receptor–G protein complexes. These observations suggest that ligand recognition, receptor activation, and transducer coupling should be viewed as integrated properties of a dynamic receptor–ligand–transducer ensemble. This perspective provides a conceptual framework that links classical GPCR pharmacology, structural biology, disease mechanisms, and next-generation drug discovery.

Pharmaceuticals doi: 10.3390/ph19081311

Authors: Kunal Verma Jaskiran Kaur Mohit Kumar Ankit Awasthi Dinesh Kumar Neeraj Choudhary Emad M. Abdallah

Background/Objectives: The aim of the present study was to investigate the mechanisms in Parkinson’s disease (PD), a progressive neurodegenerative disorder characterized by loss of dopaminergic neurons, aggregation of α-synuclein, mitochondrial dysfunction, oxidative stress, neuroinflammation, gut dysbiosis, and blood–brain barrier (BBB) impairment. Although there are several approved therapies that have been developed, their aqueous solubility, oral bioavailability, first-pass metabolism, and inability to penetrate the BBB make them less effective over time. This review is intended to critically analyze the potential of self-nanoemulsifying drug delivery systems (SNEDDSs) as a pathogenesis-related approach to enhance the delivery and therapeutic activity of repurposed drugs and conventional drugs for PD. Methods: A comprehensive literature search was conducted to address the pathogenic mechanisms of PD, the deficiencies of current pharmacotherapy, recent developments in SNEDDS formulation strategies and their application in improving oral bioavailability, lymphatic transport, BBB penetration and targeted brain delivery. A special focus was dedicated to drug repurposing, functionalized SNEDDSs, PEGylation, and gut–brain axis modulation. Results: SNEDDSs significantly enhance the water solubility, stability, intestinal absorption and systemic exposure of poorly water-soluble therapeutic agents and, to a certain extent, lymphatic uptake to avoid first-pass metabolism. These systems include improved brain delivery, decreased pharmacokinetic variability, and prolonged drug levels within the therapeutic range. Moreover, SNEDDSs can be used to deliver multiple molecules that are found to be neuroprotective, antioxidant, anti-inflammatory and probiotic, all at once, which can act on multiple pathogenic mechanisms associated with PD. Functionalized and PEGylated SNEDDSs add further to formulation stability, extend systemic circulation and increase efficiency of brain targeting. Conclusions: SNEDDSs are a promising translational nanomedicine platform for enhancing the effectiveness of conventional and repurposed therapeutics in PD, which address key pharmacokinetic and biological challenges. The next generation of oral therapies with targeted surface engineering, precision drug repurposing and clinical validation will be expected to bring about a faster advancement of drugs that can alter the course of disease rather than giving only symptomatic relief.

Pharmaceuticals doi: 10.3390/ph19081310

Authors: Marina Bilotta Adriana Gargano Roberta Rocca Valentina Maggisano Stefania Bulotta Stefano Alcaro

Background: Tankyrase 1 (TNKS1) is a poly(ADP-ribose) polymerase involved in Wnt/β-catenin signaling, telomere maintenance, and genomic stability, making it an attractive therapeutic target in oncology. This study aimed to develop and apply an integrated computational workflow to identify novel TNKS1 inhibitor candidates. Methods: A curated dataset of experimentally validated TNKS1 inhibitors and property-matched DUD-E decoys was used to develop a consensus supervised machine learning (ML) model prioritization framework for ligand-based virtual screening, integrating Morgan fingerprints with three complementary classifiers. The model screened more than 700,000 compounds, and prioritized hits were evaluated by structure-based virtual screening (SBVS), Prime MM-GBSA binding free-energy refinement, and 500 ns molecular dynamics simulations (MDs). The top candidates were subsequently tested in an in vitro TNKS1 enzymatic inhibition assay. Results: The consensus ML framework prioritized 670 compounds, yielding five candidates for experimental testing. Compound 3 displayed the most favorable computational profile and was experimentally confirmed as a TNKS1 inhibitor candidate, exhibiting approximately 80% TNKS1 inhibition at 0.1 μM, whereas the remaining candidates showed only limited activity. Conclusions: The proposed workflow efficiently reduced a large chemical space to a focused set of TNKS1 inhibitor candidates while substantially reducing the experimental screening burden. Compound 3 represents a promising starting point for future structure–activity relationship studies and lead optimization in the context of TNKS1 inhibition. Moreover, this work highlights the value of integrating consensus ML, SBVS, and experimental validation to accelerate early-stage hit discovery for TNKS1 and other therapeutic targets.

Pharmaceuticals doi: 10.3390/ph19081309

Authors: Muhammad Ishfaq Shahi Jahan Shah Imran Khalid Mashail M. M. Hamid Muhammad Zahir Kota Abdul Ahad Ghaffar Khan Mohammed Ibrahim Samuel Ebele Udeabor Abosofyan Salih Atta Elfadeel Mohamed Salih Chidozie Ifechi Onwuka

Background: Mitogen-activated protein kinase 3 (MAPK3/ERK1) plays a central role in cellular proliferation, inflammation, apoptosis, and survival signalling and has been implicated in oral squamous cell carcinoma (OSCC), periodontitis, oral lichen planus, and other chronic oral inflammatory diseases. The present study employed an integrated computational workflow combining machine learning (ML)-based quantitative structure–activity relationship (QSAR) modelling, molecular docking, density functional theory (DFT), molecular dynamics (MD) simulation, and MM-GBSA analysis to identify and characterise potent MAPK3 inhibitors. Methods: A curated dataset of 907 experimentally validated MAPK3 inhibitors was retrieved from the ChEMBL database and processed using molecular descriptors and Morgan fingerprints. Multiple ML algorithms were evaluated under scaffold-based validation, with Light Gradient Boosting Machine (LightGBM) demonstrating the best predictive performance. Results: The final model achieved strong classification capability with ROC-AUC values of 0.898 and 0.926. Feature importance analysis revealed that local structural motifs captured by fingerprint descriptors played dominant roles in MAPK3 inhibitory activity. The top-ranked compounds were subjected to molecular docking, where compounds 58324148 and 137531515 exhibited strong binding affinities of −11.9 and −11.0 kcal/mol, respectively. DFT calculations demonstrated favourable electronic properties with low HOMO–LUMO energy gaps, while MD simulations confirmed stable receptor–ligand interactions throughout 200 ns trajectories. MM-GBSA analysis further supported strong binding stability dominated by van der Waals interactions. Conclusions: Overall, the integrated computational framework successfully identified promising MAPK3 inhibitor candidates with potential therapeutic relevance for oral inflammatory and proliferative diseases.

Pharmaceuticals doi: 10.3390/ph19081308

Authors: Engi Algharably Ursula Gundert-Remy

Optimizing pharmacotherapy is a central goal of clinical pharmacology [...]

Pharmaceuticals doi: 10.3390/ph19081307

Authors: Maha Abdullah Alwaili Nawal Al-Hoshani Huda A. Alqahtani Rasha Alonaizan Khaled Alzhrani Tariq Aziz

Background/Objectives: Batai orthobunyavirus (BATV) is an emerging mosquito-borne zoonotic pathogen for which no licensed vaccine is currently available. The viral envelope glycoprotein plays an important role in viral attachment and host immune recognition, making it a potential target for rational vaccine design. Methods: In this study, an immunoinformatics-based framework was used to design and evaluate a multi-epitope vaccine candidate targeting the BATV envelope glycoprotein. Selected B-cell, cytotoxic T-lymphocyte (CTL), and helper T-lymphocyte (HTL) epitopes were assembled using appropriate linkers and a human β-defensin adjuvant. Population coverage and in silico immune simulations were conducted to evaluate the potential breadth and magnitude of immune response. Results: The final vaccine construct demonstrated favorable physicochemical characteristics, high predicted antigenicity (0.7959), and non-allergenic properties while maintaining favorable predicted structural characteristics and broad predicted population coverage (99.92%). Structural docking revealed a stable interaction between the vaccine construct and human TLR4, with a weighted docking score of −1194.8, suggesting favorable molecular recognition and receptor engagement. Normal Mode Analysis further supported the structural stability and conformational integrity of the vaccine receptor complex. Immune simulation predicted robust primary and secondary immune responses characterized by elevated IgM and IgG antibody production, sustained memory cell formation, and strong IFN-γ and IL-2 responses, indicating the potential to elicit balanced humoral and cellular immunity. Conclusions: This study presents a structurally optimized and validated multiepitope vaccine candidate against the emerging Batai orthobunyavirus. These computational findings identified a promising vaccine candidate for further investigation; however, its immunogenicity, safety, and protective efficacy before further vaccine development can be considered.

Pharmaceuticals doi: 10.3390/ph19081306

Authors: Md. Amdadul Huq Md. Ashikur Rahman Md. Rasel Rana Jong-Whi Park

The biosynthesis of copper and copper oxide nanoparticles (Cu/CuO-NPs) has attracted considerable interest due to its non-toxic, eco-friendly nature and wide-ranging applications, especially in nanomedicine and biomedical fields. Traditional nanoparticle production methods often involve toxic chemicals and generate harmful byproducts. In contrast, biological synthesis provides a cleaner, safer, more cost-effective, and sustainable alternative. Various biological sources, including plants, bacteria, fungi, and yeast, have been employed for the efficient and non-toxic production of Cu/CuO-NPs. These organisms contain diverse biomolecules such as enzymes, proteins, amino acids, vitamins, flavonoids, and alkaloids that function as reducing, capping, and stabilizing agents during nanoparticle formation. The biologically synthesized Cu/CuO-NPs are characterized using UV-VIS spectroscopy, Raman spectroscopy, TEM, SEM, EDX, XRD, TGA, XPS, FTIR, DLS, zeta potential analyzer, etc. Cu/CuO-NPs hold promise for applications in nanomedicine, primarily because of their strong antimicrobial and anticancer activities and potential use as disinfectants against infectious diseases. Various reports have suggested that the biologically synthesized Cu/CuO-NPs have exhibited significant antimicrobial and anticancer efficacies against pathogenic bacteria, fungi and viruses and various cancer cells. Due to their nanoscale dimensions and extensive surface area, Cu/CuO nanoparticles can readily infiltrate cell walls, disrupt membrane integrity, generate reactive oxygen species, and hinder both DNA replication and protein production, leading to cell death. The present review comprehensively describes the biological synthesis of Cu/CuO-NPs, their characterization techniques, and potential antibacterial, antifungal, antiviral, and anticancer applications. The modes of action for antibacterial, antifungal, antiviral, and anticancer properties have also been explored critically.

Pharmaceuticals doi: 10.3390/ph19081305

Authors: Francisco Arias-Aragón Carmen Mata-Martín Alba Sánchez-Redondo Jesús Novalbos Miguel Ángel Seguido Francisco Abad-Santos Mar Hernaez Mercè Brunet David Hansoe Heredero-Jung Alejandro de la Sota-Pérez María Isidoro-García Almudena Gil-Rodriguez Alba Barral-Raña Olalla Maroñas Gladys Guadalupe Olivera Pasquini Enrique G. Zucchet María José Herrero José Manuel Dodero-Anillo Alicia Alba Máñez María José Pedrosa-Martínez Adrián Llerena on behalf of the BioFRAM PGx Consortium

Background/Objectives: Pharmacogenetics (PGx) may improve drug safety and effectiveness, but its integration into routine care remains heterogeneous across the Spanish National Health System (NHS). This article presents BioFRAM PGx, a multicentre implementation framework that defines the methodological and operational basis for subsequent observational validation in cardiovascular and mental-health care settings. Methods: The framework was developed by healthcare centres from eight Spanish Autonomous Communities through a structured review of PharmGKB/ClinPGx clinical annotations, CPIC and DPWG guidelines, and national regulatory resources. Gene–drug pairs were prioritized according to clinical actionability, therapeutic relevance, applicability to the Spanish population, and analytical feasibility. BioFRAM PGx defines a panel of seven pharmacogenes and 35 drugs, standardized genotyping and phenotype-assignment procedures, pharmacovigilance and clinical variables, healthcare-resource measures, and centralized data management. The proposed non-interventional validation phase includes adults receiving at least one selected drug in hospital or primary-care settings, with a minimum six-month follow-up. PGx results are not returned to clinicians and do not modify treatment. Its primary objective is to assess the predictive value of PGx genotyping for adverse drug reactions (ADRs); secondary objectives include ADR incidence and implementation feasibility. Results: The main outputs are the proposed multicentre observational design, the prioritized gene–drug panel, harmonized analytical and pharmacovigilance workflows, standardized data domains, and a centralized data-management strategy. No patient-level clinical, implementation, or economic outcomes are presented. Conclusions: BioFRAM PGx provides a structured multicentre framework for harmonizing pharmacogenetic procedures across the Spanish NHS. Its clinical effectiveness, influence on prescribing, scalability, and economic outcomes remain to be assessed in subsequent studies.

Pharmaceuticals doi: 10.3390/ph19081304

Authors: Sara Shefa Jagoda Szwach Anna Karwowska Aleksandra Wojno Magdalena Łyko

Acne vulgaris is one of the most prevalent dermatological conditions, affecting up to 85% of individuals aged 11–30 years. Clascoterone cream is the first topical androgen receptor inhibitor approved by the U.S. Food and Drug Administration (FDA) for the treatment of acne vulgaris in patients aged 12 years and older, and the first agent in this class approved for use in both male and female patients. The aim of this narrative review is to summarize the current evidence regarding the efficacy, safety profile, and potential future applications of clascoterone in the treatment of acne vulgaris. A comprehensive literature search was conducted in PubMed/MEDLINE and ClinicalTrials.gov databases up to April 2026. Search terms included “clascoterone,” “cortexolone 17α-propionate,” and “androgen receptor inhibitor acne.” Eligible publications included randomized controlled trials, meta-analyses, open-label extension studies, retrospective analyses, case reports, and relevant review articles published in English. Clinical trial registries and regulatory agency announcements were also searched for ongoing studies and recent approval decisions. Phase 3 trials and meta-analyses demonstrate that twice-daily application of clascoterone 1% cream significantly reduces inflammatory and non-inflammatory lesion counts compared to vehicle, with efficacy sustained up to 12 months of treatment. The safety profile is favorable, with adverse events limited predominantly to mild and transient local skin reactions. Subgroup analyses have shown no significant sex-related differences in efficacy or safety, and the safety profile appears comparable between adolescents and adults. In an exploratory pooled subgroup analysis, lesion-count reductions were greater in adults than in adolescents, whereas the adjusted between-group comparison of IGA success was not statistically significant. Because no formal treatment-by-age interaction test was reported, these findings should be considered hypothesis-generating. Emerging but still incomplete data suggest potential applications in androgenetic alopecia and mild hidradenitis suppurativa. Clascoterone represents a novel addition to the therapeutic armamentarium for acne vulgaris, offering a unique mechanism of action with a favorable safety profile. Further research is warranted to establish its role in combination therapies and in other androgen-dependent dermatoses.