Pharmaceuticals

22 pages, 5196 KB

Open AccessArticle

Phytotherapeutic Intervention in Monosodium Glutamate-Induced Uterine Dysfunction: Efficacy of Lepidium sativum, Prunus armeniaca, Stachys palustris, and Solenostemma argel

by Eslam ElNebrisi, Nadia M. El Rouby, Fatimah Muaamar Noori, Nikoo Ali Jalali, Rodiana Mohamed Fouad Saber, Zainab Safieldin Abdalla Mohamed and Naglaa Gamil Shehab

Pharmaceuticals 2026, 19(3), 521; https://doi.org/10.3390/ph19030521 - 23 Mar 2026

Viewed by 767

Abstract

Introduction: Uterine fibroids are benign tumors arising from uterine smooth muscle and significantly affect women’s health worldwide. While conventional treatments often involve hormonal therapies or invasive surgeries, these approaches are limited by cost, side effects, and fertility concerns. This study aimed to [...] Read more.

Introduction: Uterine fibroids are benign tumors arising from uterine smooth muscle and significantly affect women’s health worldwide. While conventional treatments often involve hormonal therapies or invasive surgeries, these approaches are limited by cost, side effects, and fertility concerns. This study aimed to evaluate the in vivo bioactivity of four medicinal plant extracts, Lepidium sativum, Prunus armeniaca, Solenostemma argel, and Stachys palustris, in ameliorating monosodium glutamate (MSG)-induced uterine changes in rats, providing preliminary preclinical evidence. Methods: The extracts were evaluated for their flavonoid and total phenolic contents, antioxidant capacity, and hormonal modulatory effects. Female Wistar rats were treated with monosodium glutamate to induce uterine changes, followed by interventions with herbal extracts. Outcomes were evaluated via biochemical, hormonal, and histological analyses. Results: Among the four extracts, Lepidium sativum and Stachys palustris showed superior antioxidant activity, restoring catalase, glutathione, and superoxide dismutase levels. These extracts also significantly reduced estrogen levels and estrogen receptor expression, correlating with improved histological outcomes, including reduced endometrial hyperplasia and myometrial thickness. Solenostemma argel and Prunus armeniaca exhibited moderate effects. Conclusions: This study underscores the potential of Lepidium sativum and Stachys palustris as natural therapeutic agents for fibroid management through antioxidant activity and hormonal modulation. Future research should focus on clinical validation to translate these findings into effective treatments. Full article

(This article belongs to the Special Issue Natural Products and Phytomedicines: From Chemical Profiling and Pharmacological Properties to Therapeutic Applications)

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28 pages, 31042 KB

Open AccessArticle

Danggui Buxue Decoction and Its Active Constituents Inhibit Drug-Induced Uterine Contractions via L-Type Calcium Channels and the IP₃/Ca²⁺ Pathway

by Mingming Liu, Taiping He, Wenqiao An, Pengmei Guo, Tang Zhou, Yufei Chen, Xiaojuan Tian, Mingxu Wu, Ting Zhang and Sanyin Zhang

Pharmaceuticals 2026, 19(3), 520; https://doi.org/10.3390/ph19030520 - 23 Mar 2026

Viewed by 857

Abstract

Background/Objectives: Primary dysmenorrhea is a common gynecological disorder characterized by painful uterine contractions. Danggui Buxue Decoction (DBD) is used to treat menstrual irregularities, but its mechanism in primary dysmenorrhea remains unclear. This study investigated the efficacy of DBD against dysmenorrhea and its [...] Read more.

Background/Objectives: Primary dysmenorrhea is a common gynecological disorder characterized by painful uterine contractions. Danggui Buxue Decoction (DBD) is used to treat menstrual irregularities, but its mechanism in primary dysmenorrhea remains unclear. This study investigated the efficacy of DBD against dysmenorrhea and its calcium signaling-related mechanism. Methods: DBD components were analyzed by UPLC–Orbitrap MS. Isolated uterine muscle strips precontracted with oxytocin (OT, 50 ng/mL) or KCl (60 mM) were used to assess the effects of DBD and its active compounds (Quercetin, Formononetin, Ononin, Ferulic acid, Senkyunolide I, Calycosin, Ligustilide, Calycosin-7-O-β-D-glucoside). Ca²⁺-dependent experiments, intracellular calcium release assays, and inhibitor treatments (Nifedipine, 2-APB) were performed to evaluate the involvement of L-type calcium channels and the IP₃R pathway. A primary dysmenorrhea model induced by estradiol benzoate and oxytocin was used to assess the analgesic effects, histopathology, inflammatory factors, and IP₃/Ca²⁺-related proteins and genes following DBD and Quercetin treatment. Results: A total of 161 compounds were identified in DBD. DBD and its eight active constituents relaxed OT (50 ng/mL) or KCl (60 mM)-induced uterine contractions, with Quercetin, Calycosin, and Ligustilide showing particularly prominent relaxant activity. These three compounds suppressed extracellular calcium influx and intracellular calcium release through the blockade of L-type calcium channels and IP₃R. In vivo, DBD and Quercetin alleviated pain, reduced inflammation, and decreased uterine Ca²⁺ and IP₃ levels in dysmenorrhea mice. Conclusions: DBD and its active component Quercetin promote uterine relaxation by lowering Ca²⁺ levels, which is achieved through suppression of L-type calcium channels and the IP₃/Ca²⁺ pathway. This contributes to their therapeutic action against primary dysmenorrhea. Full article

(This article belongs to the Special Issue Advances in Smooth Muscle Pharmacology)

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68 pages, 6786 KB

Open AccessReview

Pleiotropic Bioactivity of Caterpillar Fungus, Orange Cordyceps, and Cordycepin: Insight from Integrated Network Pharmacology and Food and Drug Regulatory Framework

by Alexander Panossian

Pharmaceuticals 2026, 19(3), 519; https://doi.org/10.3390/ph19030519 - 23 Mar 2026

Viewed by 1375

Abstract

Background/Objectives: The medical mushroom Ophiocordyceps sinensis (Caterpillar Fungus), known for its ability to enhance “vitality,” is one of the most popular medicines in Asian traditional medical systems. According to the Chinese Pharmacopeia, O. sinensis is standardized for its adenosine content, the precursor [...] Read more.

Background/Objectives: The medical mushroom Ophiocordyceps sinensis (Caterpillar Fungus), known for its ability to enhance “vitality,” is one of the most popular medicines in Asian traditional medical systems. According to the Chinese Pharmacopeia, O. sinensis is standardized for its adenosine content, the precursor of ATP, which mediates numerous physiological and pathological processes in many diseases. The related fungus of order Hypocreales, Cordyceps militaris, and its major bioactive constituents, 3′-deoxyadenosine (cordycepin), also exhibit pleiotropic biological activities. This review aims to provide a rationale for the adaptogenic and resilience-supporting effects of these medicinal fungi and to align food and drug regulation in Western countries. Methods: In this narrative review, we integrated results from chemical, pharmacokinetic, network pharmacology, preclinical, and clinical studies of O. sinensis, C. militaris, and cordycepin using network pharmacology and bioinformatics tools. Results: Across studies, recurrent mechanistic hubs included PI3K–Akt, AMPK–mTOR, MAPK, NF-κB, apoptosis, and adaptive stress-response signaling pathways, linking immune regulation and metabolic homeostasis. Experimental studies confirmed modulation of cytokine production, kinase signaling, and mitochondrial regulators. Clinical meta-analyses demonstrate consistent adjunctive benefits in renal and pulmonary disorders, although heterogeneity in preparation and methodological limitations remains significant. The review reveals controversy regarding the bioavailability of cordycepin in vivo and its concentration in vitro studies, raising the hypothesis that cordycepin may act as a driver, triggering the organism’s adaptive stress response in stress-induced and aging-related diseases. Pharmacokinetic data indicate that systemic cordycepin concentrations after oral administration remain in the nanomolar range, suggesting that some predicted molecular interactions may occur indirectly or through systems-level mechanisms. The review, for the first time, suggests establishing a regulatory category for resilience-supporting physiological modulators to align food and drug regulation in the EU with contemporary systems biology, thereby complementing the work of EFSA, EMA, FDA, and Asian authorities. Conclusions:O. sinensis, C. militaris, and 3-deoxyadenosine share a common adaptogenic mechanism for maintaining homeostasis of cellular and integrated biological system functions. The systems-level network analysis and reductionistic molecular ligand preceptor pharmacology provide complementary approaches for understanding the multi-target bioactivity of these fungi. This review clarifies conceptual and regulatory barriers to recognizing resilience-supporting interventions and informs future regulatory innovation. Full article

(This article belongs to the Special Issue Network Pharmacology of Natural Products, 2nd Edition)

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12 pages, 482 KB

Open AccessArticle

Bioabsorbable Hydrogel Coating for Infection Prevention in Fracture Fixation: A Retrospective Matched Case–Control Study

by Carlo Ciccullo, Marco Grassi, Marco Antonio Carletti, Claudia Bevilacqua, Danilo Francesco Chirillo, Simone Domenico Aspriello and Antonio Pompilio Gigante

Pharmaceuticals 2026, 19(3), 518; https://doi.org/10.3390/ph19030518 - 23 Mar 2026

Viewed by 590

Abstract

Background/Objectives: Hospital-acquired and fracture-related infections remain major complications in orthopedic trauma surgery, with significant clinical and socio-economic impact. Antibacterial implant surface coatings represent a promising strategy to reduce early postoperative bacterial adhesion and biofilm formation. Methods: This retrospective matched case–control study [...] Read more.

Background/Objectives: Hospital-acquired and fracture-related infections remain major complications in orthopedic trauma surgery, with significant clinical and socio-economic impact. Antibacterial implant surface coatings represent a promising strategy to reduce early postoperative bacterial adhesion and biofilm formation. Methods: This retrospective matched case–control study evaluated the clinical effectiveness of an antibiotic-free fast-resorbable hyaluronic acid and poly-d, l-lactide hydrogel (DAC^®) applied intraoperatively to orthopedic implants. A total of 222 patients with comorbidities who underwent open reduction and internal fixation between May 2023 and April 2024 in two trauma centers were included: 99 patients received the DAC^® coating and 123 served as controls with standard fixation. The primary endpoint was infection incidence within 6 months; secondary endpoints included wound complications, revision surgery, prolonged antibiotic therapy, and bone healing. Results: Postoperative infection incidence was significantly lower in the DAC^® group compared with controls (0.7% vs. 5.3%; p = 0.0363). Wound complications were also reduced (1.3% vs. 8.0%; p = 0.028), and only one patient in the DAC^® cohort required additional surgical interventions or prolonged antibiotic therapy. Bone healing outcomes were comparable between groups, with no delayed unions reported in the treated cohort. Conclusions: Even if larger prospective studies with longer follow-up are required to further confirm these findings and better define long-term safety and effectiveness, the routine intraoperative use of DAC^® hydrogel without antibiotic loading appears to be a safe and promising strategy to reduce early postoperative infections and wound complications in orthopedic trauma patients with comorbidities. Full article

(This article belongs to the Topic Applications of Polymers and Polymer Nanomaterials in Drug Delivery and Nanomedicine, 2nd Edition)

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16 pages, 2460 KB

Open AccessArticle

Enhanced Tumor-to-Background Contrast with [⁵²Mn]Mn-BPPA-Bevacizumab VEGF-Targeted Immuno-PET in Cervical Cancer

by Csaba Csikos, Minh Toàn Ngô, Adrienn Vágner, Gábor Nagy, Gábor Ország, Tamás Nagy, Balázs Váradi, Gergő Zoltán Sajtos, István Kapus, Zoltán Szoboszlai, Dezső Szikra, Gyula Tircsó, Zoárd Tibor Krasznai, Szabolcs Molnár, Ildikó Garai and György Trencsényi

Pharmaceuticals 2026, 19(3), 517; https://doi.org/10.3390/ph19030517 - 22 Mar 2026

Viewed by 961

Abstract

Background/Objectives: Radiolabeled bevacizumab-based immuno-PET tracers enable a non-invasive quantification of VEGF-A expression in gynecologic malignancies. While the previously reported [⁵²Mn]Mn-DOTAGA-bevacizumab demonstrated selective VEGF-A-targeted uptake in a KB-3-1 cervix carcinoma mouse model, further improvements in chelator stability and tumor-to-background contrast remain [...] Read more.

Background/Objectives: Radiolabeled bevacizumab-based immuno-PET tracers enable a non-invasive quantification of VEGF-A expression in gynecologic malignancies. While the previously reported [⁵²Mn]Mn-DOTAGA-bevacizumab demonstrated selective VEGF-A-targeted uptake in a KB-3-1 cervix carcinoma mouse model, further improvements in chelator stability and tumor-to-background contrast remain desirable. The recently developed BPPA chelator exhibits exceptionally high Mn(II) complex stability and favorable radiolabeling characteristics. This study aimed to characterize the in vivo biodistribution of [⁵²Mn]Mn-BPPA-bevacizumab, and to compare the tumor-to-background ratios of [⁵²Mn]Mn-BPPA-bevacizumab with the previously published values of [⁵²Mn]Mn-DOTAGA-bevacizumab in VEGF-A-expressing cervix carcinoma. Methods: Female KB-3-1 tumor-bearing CB17 SCID mice underwent PET/MRI imaging following intravenous administration of [⁵²Mn]Mn-BPPA-bevacizumab. SUV_mean values were measured in various organs and in the subcutaneously injected tumor, and tumor-to-organ ratios were calculated at various time points up to 10 days post-injection. Results: [⁵²Mn]Mn-BPPA-bevacizumab demonstrated sustained tumor uptake, with tumor SUVmean values increasing from approximately 1.0 at 4 h to peak values of approximately 2.4–2.5 at 72 h post-injection. Tumor-to-background ratios increased progressively over time and were significantly higher for [⁵²Mn]Mn-BPPA-bevacizumab compared with previously reported [52Mn]Mn-DOTAGA-bevacizumab, particularly for tumor-to-blood, tumor-to-liver and tumor-to-lung ratios at later imaging time points (p < 0.0001). Conclusions: The novel [⁵²Mn]Mn-BPPA-bevacizumab tracer exhibits satisfactory in vitro and in vivo stability for PET imaging, high VEGF-A-specific tumor uptake, and markedly improved tumor-to-background ratios compared to the previously published DOTAGA-based probe. These results position [⁵²Mn]Mn-BPPA-bevacizumab as a highly promising next-generation immuno-PET agent for imaging VEGF-A-expressing gynecologic malignancies and for guiding anti-angiogenic therapies. Full article

(This article belongs to the Section Radiopharmaceutical Sciences)

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11 pages, 1153 KB

Open AccessArticle

Efficacious Anti-Cancer Drugs Targeting Nicotinamide N-Methyltransferase (NNMT) in Cultured Human Oral Squamous Cell Carcinoma (OSCC)

by Brian Maloney, Martyna Kubisztal, Ziqian Ge, Yin Lu, Lisa Strotmann, Adrianna Budziňska, Mary F. Rooney, Marilena Karavyraki, Andrew Knox and Richard K. Porter

Pharmaceuticals 2026, 19(3), 516; https://doi.org/10.3390/ph19030516 - 22 Mar 2026

Viewed by 781

Abstract

Background/Objectives: Oral squamous cell carcinoma (OSCC) is a major cause of human cancer. The enzyme, nicotinamide N-methyltransferase (NNMT), is overexpressed in a variety of human cancers, including OSCC. Our objective was to target NNMT with novel inhibitors and determine their anti-cancer efficacy [...] Read more.

Background/Objectives: Oral squamous cell carcinoma (OSCC) is a major cause of human cancer. The enzyme, nicotinamide N-methyltransferase (NNMT), is overexpressed in a variety of human cancers, including OSCC. Our objective was to target NNMT with novel inhibitors and determine their anti-cancer efficacy while shedding light on their possible mechanism of action. Methods: We identified two small molecule inhibitors of NNMT (AG-670 and AO-022) based on a pharmacophore of the in silico nicotinamide binding site. These inhibitors were investigated for (i) potency to inhibit the activity of the isolated NNMT enzyme (EC₅₀ values), (ii) cytotoxicity (IC₅₀ values) against the human OSCC cell line, SCC-4, and (iii) ability to affect cellular energy metabolism, as measured by oxygen consumption, in SCC-4 cells (plus dysplastic oral keratinocytes (DOK) cells and breast cancer MCF-7 cells). Immunoblotting was used to determine whether NNMT was expressed in the aforementioned cells. Results: NNMT is expressed in SCC-4 and DOK cells (and primary human oral keratinocytes) but not MCF 7 cells. The NNMT inhibitors inhibit isolated NNMT enzyme activity and were cytotoxic to SCC-4 cells (EC₅₀ and IC₅₀ values in the micromolar range). Sublethal doses of the inhibitors were demonstrated to inhibit in situ mitochondrial oxygen consumption in SCC-4 and DOK cells but not in MCF-7 cells. It was demonstrated that the NNMT inhibitors do not directly inhibit mitochondrial electron transport chain activity. Thus, we deduce that the NNMT inhibitors affect mitochondrial activity indirectly via NNMT. Conclusions: It is concluded that NNMT is a potential drug target for oral cancer. Full article

(This article belongs to the Section Medicinal Chemistry)

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22 pages, 2041 KB

Open AccessArticle

Rational Design, Synthesis, and Systematic Evaluation of Redox-Responsive SN-38 Prodrugs for Selective Activation in Hypoxic Tumor Microenvironments

by Taimin Dong, Jin Xu, Xiuling Wang, Ziqiao Sun, Shuo Wang, Fanghui Chen, Hanchuang Zhu, Xinyu Zhang, Shuhai Xu, Chunguang Zheng, Dan Mao, Tianying Ren, Qiaoling Ni, Chenjing Xu, Xinyi Shen, Na Li, Dapeng Zhang, Lusha Ji, Huaizu Guo and Xuekun Wang

Pharmaceuticals 2026, 19(3), 515; https://doi.org/10.3390/ph19030515 - 21 Mar 2026

Viewed by 676

Abstract

Background: The potent topoisomerase I inhibitor SN-38, the active metabolite of irinotecan, is limited in clinical application due to severe systemic toxicity. Prodrug strategies enabling selective activation in the tumor microenvironment offer a promising approach to improve its therapeutic index. This study aims [...] Read more.

Background: The potent topoisomerase I inhibitor SN-38, the active metabolite of irinotecan, is limited in clinical application due to severe systemic toxicity. Prodrug strategies enabling selective activation in the tumor microenvironment offer a promising approach to improve its therapeutic index. This study aims to rationally design, synthesize, and systematically evaluate novel disulfide-based SN-38 prodrugs engineered for redox-responsive activation in hypoxic tumors. Methods: Two novel disulfide-based SN-38 prodrugs (SN-38-CSS and SN-38-LSS) were designed and synthesized; SN-38-CSS incorporates a constrained cis-piperazine-fused six-membered cyclic disulfide linker, while SN-38-LSS contains a linear disulfide tether, to differentially exploit the upregulated thioredoxin (Trx/TrxR) system in hypoxic tumor microenvironments. Results: Both prodrugs demonstrated high stability under physiological pH conditions and in human plasma, minimizing premature release. Crucially, they exhibited selective, rapid degradation in the presence of dithiol reductants (TCEP and DTT), mimicking Trx system activity, while remaining stable towards monothiols (GSH, L-Cys). In vitro cytotoxicity assays revealed that the prodrugs exhibited significantly reduced toxicity compared to SN-38 under normoxic conditions across most tested cell lines. However, under hypoxic conditions, their activity was significantly restored. Specifically, SN-38-CSS exhibited cytotoxicity comparable to SN-38 against MCF-7 and NCI-N87 cells, whereas SN-38-LSS showed lower activation efficiency. Conclusions: SN-38-CSS is identified as a promising redox and hypoxia dual-responsive prodrug candidate, highlighting the strategic use of cyclic disulfide linkers for achieving high selectivity and controlled drug release within the tumor microenvironment. Full article

(This article belongs to the Section Medicinal Chemistry)

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18 pages, 1920 KB

Open AccessArticle

Pharmacogenetic Associations with Statin Regimen Modification, Intolerance, and Adverse Outcomes in Coronary Artery Disease Patients

by Rania Abdel-latif, Shaban Mohammed, Mohamad Saad, Khalid Kunji, Wadha Al-Muftah, Ayman El-Menyar and Jassim Al Suwaidi

Pharmaceuticals 2026, 19(3), 514; https://doi.org/10.3390/ph19030514 - 21 Mar 2026

Viewed by 638

Abstract

Background: Statins are central to primary and secondary prevention of atherosclerotic cardiovascular disease but are often underutilized due to myopathy and intolerance. While individual pharmacogenetic (PGx) variants, particularly in SLCO1B1, are linked to statin-associated muscle symptoms, the real-world impact of both [...] Read more.

Background: Statins are central to primary and secondary prevention of atherosclerotic cardiovascular disease but are often underutilized due to myopathy and intolerance. While individual pharmacogenetic (PGx) variants, particularly in SLCO1B1, are linked to statin-associated muscle symptoms, the real-world impact of both clinical and cumulative PGx burden on regimen modification and adverse outcomes remains unclear. We aimed to evaluate the existing uncertainty regarding whether combined PGx scores can effectively guide statin dose titration and regimen modification, thereby filling a key clinical gap. Methods: A retrospective cohort study of 911 statin-treated patients with coronary artery disease was conducted from the Qatar Cardiovascular Biorepository with available whole-genome sequencing data. Variants in SLCO1B1, ABCG2, and CYP2C9 were combined into a functional PGx burden score, and their associations with statin regimen modification, intolerance, myopathy, liver injury, adherence, and composite adverse events were evaluated. The composite adverse events were defined as the occurrence of any statin-related adverse event, including statin-associated myopathy, liver injury, or poor medication adherence, during the follow-up period. Patients were classified as having experienced the composite outcome if at least one of these events occurred. Results: Over 12 months following statin initiation, 10.2% of patients underwent dose escalation, 11.4% de-escalation, and 78.4% remained on the same regimen. PGx burden is not statistically significantly associated with statin intolerance (OR 1.14; 95% CI: 0.73–1.76), composite adverse outcome (OR 1.08; 95% CI 0.82–1.42), or time to regimen change (HR 1.02; 95% CI 0.77–1.35). However, higher PGx burden showed a directional tendency toward dose de-escalation (RRR 1.18, 95% CI 0.76–1.84) and lower likelihood of escalation (RRR 0.93, 95% CI 0.56–1.54). Conclusions: Clinical factors, particularly statin intensity and myopathy, were the primary determinants of regimen modification. The PGx burden contributes to vulnerability to statin-related adverse effects in a context-dependent manner but does not independently drive statin regimen modification in routine clinical practice. Prospective studies are warranted to assess the clinical utility of PGx-guided workflows in statin therapy. Full article

(This article belongs to the Special Issue Pharmacovigilance Insights: Addressing Drug Misuse and Enhancing Drug Safety)

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17 pages, 832 KB

Open AccessArticle

Central and Peripheral Characterization of Key Kynurenine Pathway Metabolites in Mexican Patients with Multiple Sclerosis: An Exploratory Study

by Pablo Arturo Acosta Mendez, Graciela Ordoñez, Karla F. Meza-Sosa, Tonali Blanco Ayala, Daniela Ramirez Ortega, Gonzalo Pérez de la Cruz, Dinora F. González Esquivel, Teresita Corona, José Flores Rivera, Verónica Rivas, Paul Carrillo Mora, Carmen Aláez-Verson, Korrapati V. Sathyasaikumar, Saúl Gomez-Manzo, Aleli Salazar, Benjamin Pineda and Verónica Pérez de la Cruz

Pharmaceuticals 2026, 19(3), 513; https://doi.org/10.3390/ph19030513 - 21 Mar 2026

Viewed by 880

Abstract

Background/Objectives: Multiple Sclerosis (MS) is a chronic immune-mediated disorder characterized by neuroinflammation and neurodegeneration. Increasing evidence implies the kynurenine pathway (KP) in the MS pathophysiology; however, data from Mexican populations are lacking. This exploratory study aimed to characterize central and circulating KP [...] Read more.

Background/Objectives: Multiple Sclerosis (MS) is a chronic immune-mediated disorder characterized by neuroinflammation and neurodegeneration. Increasing evidence implies the kynurenine pathway (KP) in the MS pathophysiology; however, data from Mexican populations are lacking. This exploratory study aimed to characterize central and circulating KP metabolites in Mexican patients with MS and to investigate potential genetic variants in KP-related genes. Methods: Serum concentrations of kynurenic acid (KYNA) and 3-hydroxykynurenine (3-HK), as well as cerebrospinal fluid (CSF) levels of KYNA, quinolinic acid (QUIN), interleukin-4 (IL-4), and interleukin-6 (IL-6), were determined in treatment-naïve relapsing-remitting MS (RRMS), primary progressive MS (PPMS), and treated PMS patients. Serum levels were compared with those of healthy controls, and CSF findings contrasted with those of non-MS neurological patients and individuals with neurocysticercosis (NCC). Public whole-exome datasets were analyzed for variants in KP-related genes, and target exome sequencing was performed in three Mexican patients with MS. Results: Serum concentrations of KYNA and 3-HK were decreased in MS patients compared with healthy controls. CSF KYNA and QUIN levels did not differ significantly among MS subtypes or the non-MS neurological group, but they were lower than those observed in NCC. IL-4 and IL-6 were detectable in MS CSF samples, supporting the presence of intrathecal inflammation. Genetic and bioinformatic analyses identified variants in genes encoding KP enzymes in both public MS datasets and in Mexican patients with MS. Conclusions: These findings indicate an altered KP metabolism in Mexican MS patients, particularly during the relapse phase, and suggest a possible contribution of genetic variability. Further large-scale studies are needed to confirm these observations and to determine the functional implications of KP-related genetic variants in MS. Full article

(This article belongs to the Special Issue Kynurenine Pathway: A Novel Therapeutic Opportunity—2nd Edition)

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19 pages, 1473 KB

Open AccessArticle

Phytochemical Composition and Bioactivity of Acanthus dioscoridis var. perringii: An Integrated Analysis of Antioxidant Activity, Enzyme Inhibition, and Phenolic–Bioactivity Correlations

by Bedrettin Selvi

Pharmaceuticals 2026, 19(3), 512; https://doi.org/10.3390/ph19030512 - 20 Mar 2026

Viewed by 476

Abstract

Objectives: Plant organs often allocate phenolic metabolites unevenly, resulting in organ-specific bioactivities. This study aimed to characterize the organ-specific phenolic profile of Acanthus dioscoridis var. perringii and determine how this chemical segregation is associated with antioxidant capacity and enzyme inhibitory activities. Materials and [...] Read more.

Objectives: Plant organs often allocate phenolic metabolites unevenly, resulting in organ-specific bioactivities. This study aimed to characterize the organ-specific phenolic profile of Acanthus dioscoridis var. perringii and determine how this chemical segregation is associated with antioxidant capacity and enzyme inhibitory activities. Materials and Methods: Organ-specific extracts (roots, stems, leaves, bracts, and flowers) were evaluated for total phenolic and flavonoid contents, targeted LC-MS analysis of individual phenolics, antioxidant activity by multiple assays, enzyme inhibition [acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, and α-glucosidase], and the relationships between phenolic composition and biological activities. Antioxidant performance was also assessed using the Relative Antioxidant Capacity Index (RACI). Results and Discussion: Roots showed the highest total phenolic content, whereas bracts had the highest total flavonoid level. Verbascoside was the dominant compound in all organs, with the highest levels in leaves, roots, and bracts. Roots exhibited the strongest reducing and radical-scavenging activities, while flowers showed the best metal-chelating capacity. Enzyme inhibition was organ-dependent and generally moderate, with stems showing the strongest cholinesterase inhibition, leaves the strongest α-amylase inhibition, and bracts together with roots the strongest α-glucosidase inhibition. Statistical analysis revealed close associations between phenolic richness, antioxidant responses, and cholinesterase inhibition. Conclusions: These findings demonstrate a clear organ-dependent distribution of phenolic compounds in A. dioscoridis var. perringii, reflected in distinct antioxidant and enzyme inhibitory profiles. Overall, the study provides a biochemical and bioactivity-based characterization of the species at the organ level. Full article

(This article belongs to the Special Issue Plant-Based Extracts and the Therapeutic Potential of Bioactive Compounds, 2nd Edition)

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18 pages, 3153 KB

Open AccessArticle

Genetic Polymorphisms Associated with Lithium Response in Bipolar Disorder: An Integrative Review and In Silico Protein–Protein Interaction Analysis

by Ovinuchi Ejiohuo and Aleksandra Szczepankiewicz

Pharmaceuticals 2026, 19(3), 511; https://doi.org/10.3390/ph19030511 - 20 Mar 2026

Viewed by 822

Abstract

Background/Objectives: Management of bipolar disorder is marked by variability in lithium response, with responders constituting a distinct clinical subgroup. Although pharmacogenetic studies implicate polymorphisms in neuroplasticity-related genes (BDNF) and hypothalamic–pituitary–adrenal (HPA) axis regulators (NR3C1), the underlying biophysical mechanisms [...] Read more.

Background/Objectives: Management of bipolar disorder is marked by variability in lithium response, with responders constituting a distinct clinical subgroup. Although pharmacogenetic studies implicate polymorphisms in neuroplasticity-related genes (BDNF) and hypothalamic–pituitary–adrenal (HPA) axis regulators (NR3C1), the underlying biophysical mechanisms remain poorly characterized. This study aims to bridge this structural–mechanistic gap by quantifying the atomic-level effects of key lithium-response polymorphisms on protein–protein interaction stability and conformational dynamics. Methods: Variant sequences for BDNF rs6265 and NR3C1 rs56149945 were generated and structurally modeled with SWISS-MODEL. Protein–protein interaction analyses focused on the BDNF–TrkB and NR3C1–FKBP5 systems. Structural alignment and conformational comparisons were performed with ChimeraX and US-align, while interaction energetics were evaluated with PRODIGY and HawkDock. Conformational flexibility was assessed using CABS-flex through RMSF analysis. Results: Structural validation showed acceptable model quality. Binding analyses indicated stronger interactions in the variant complexes. In the BDNF–TrkB complex, binding affinity shifted from −13.8 to −15.1 kcal/mol with an ~8.5-fold lower dissociation constant, while the NR3C1–FKBP5 variant complex shifted from −16.3 to −18.8 kcal/mol with an ~65-fold lower dissociation constant. MM/GBSA calculations supported increased stability, with binding energies changing from −61.98 to −83.91 kcal/mol (BDNF–TrkB) and from −18.88 to −31.25 kcal/mol (NR3C1–FKBP5). Structural superposition showed high conservation of global folds (pruned RMSD 0.779 Å and 0.310 Å; TM-scores 0.753 and 0.967). RMSF profiles were largely overlapping, indicating localized interface adjustments rather than global conformational changes. Conclusions: These findings suggest that lithium-response polymorphisms may modulate protein–protein interaction stability while preserving overall structure, providing a structural framework for exploring genetic influences on lithium treatment response. Full article

(This article belongs to the Section Pharmacology)

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23 pages, 6932 KB

Open AccessArticle

CIGB-258, a Potential Novel Approach to Treat Sepsis-like Hyperinflammation, Reduces Gastrointestinal Hemorrhage in Zebrafish Exposed to Carboxymethyllysine and Ethanol

by Kyung-Hyun Cho, Yunki Lee, Sang Hyuk Lee, Ashutosh Bahuguna, María del Carmen Domínguez-Horta and Gillian Martínez-Donato

Pharmaceuticals 2026, 19(3), 510; https://doi.org/10.3390/ph19030510 - 20 Mar 2026

Cited by 1 | Viewed by 968

Abstract

Objective: CIGB-258 is a 3 KDa altered peptide ligand recognized for its anti-inflammatory activity. Herein, the effect of CIGB-258 was assessed against carboxymethyllysine (CML) and ethanol (Et-OH)-induced sepsis-like events in zebrafish (Danio rerio). Methodology: Adult zebrafish (n = 30/group) were intraperitoneally [...] Read more.

Objective: CIGB-258 is a 3 KDa altered peptide ligand recognized for its anti-inflammatory activity. Herein, the effect of CIGB-258 was assessed against carboxymethyllysine (CML) and ethanol (Et-OH)-induced sepsis-like events in zebrafish (Danio rerio). Methodology: Adult zebrafish (n = 30/group) were intraperitoneally microinjected (10 μL) with CML (final 3 mM) + Et-OH (final 50%) or CML + Et-OH containing CIGB-258 (final 1 μM) and analyzed for swimming activity, abdominal bleeding and survivability. The zebrafish were sacrificed 180 min after injection, and blood and organs were processed for biochemical and histological evaluation. Results: The CML + Et-OH group showed the lowest survival, compromised swimming ability, and severe abdominal bleeding 60 min post-treatment, which were substantially improved by treatment with CIGB-258. The CML + Et-OH group showed the greatest extent of oxidization and the lowest antioxidant activity in plasma, while co-treatment with CIGB-258 resulted in a remarkable improvement in oxidative extent and antioxidant status. The CML + Et-OH group showed dyslipidemia and an atherogenic lipid profile, which were substantially prevented by the CIGB-258 treatment. The livers and kidneys of the CML + Et-OH group showed the greatest extent of inflammation and senescence, which were substantially ameliorated by treatment with CIGB-258. Similarly, the CML + Et-OH group exhibited severe intestinal bleeding, which decreased 2.2-fold following treatment with CIGB-258. H&E staining and Mason-trichrome staining revealed extreme disruption to intestinal microvillus cell morphology and severe fibrosis in the intestines of the CML + Et-OH group, which effects were mitigated by the treatment with CIGB-258. Conclusions: The CML + Et-OH treatment resulted in acute gastrointestinal bleeding, severe oxidative stress, and hepatic and renal damage, leading to acute septic shock-like death in zebrafish. However, treatment with CIGB-258 reduced these effects through antioxidant and anti-inflammatory actions and by increasing HDL-C levels. Full article

(This article belongs to the Special Issue Research on Diagnosis, Treatment and Related Microbiota in Gastrointestinal Diseases)

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38 pages, 4695 KB

Open AccessArticle

Potential Mechanisms of MAP Kinase JNK’s Involvement in Modulating Cancer Cell Fate in a Cisplatin Concentration-Dependent Manner

by Monika Tenkutytė, Audronė V. Kalvelytė and Aurimas Stulpinas

Pharmaceuticals 2026, 19(3), 509; https://doi.org/10.3390/ph19030509 - 20 Mar 2026

Viewed by 698

Abstract

Background: The combination of conventional drugs and inhibitors of signaling molecules is an effective strategy to increase cancer treatment efficacy and reduce drug doses to protect against their cytotoxic effects. Our research has shown the cisplatin concentration-dependent shift in the role of MAP [...] Read more.

Background: The combination of conventional drugs and inhibitors of signaling molecules is an effective strategy to increase cancer treatment efficacy and reduce drug doses to protect against their cytotoxic effects. Our research has shown the cisplatin concentration-dependent shift in the role of MAP kinase JNK from antiapoptotic to proapoptotic in non-small cell lung cancer A549 cells. Cell death/survival signaling molecules, tumor suppressor p53 and pro-survival protein kinase AKT were detected to be differently regulated by JNK inhibition at low vs. high cisplatin concentrations. Here, we further investigated the phenomenon and potential mechanisms of combined JNK inhibition and cisplatin treatment. Methods: Cell death in vitro was evaluated by MTT and Western blot assays after combined cisplatin and specific inhibitor treatment; two-way ANOVA was used for analysis. Results: JNK is differently involved in determining cellular sensitivity to different DNA-damaging drugs. There is no universal cell death induction mechanism originating from DNA damage through the involvement of JNK. The outcome of JNK inhibition also depends on the cell type. We found that there is an unusual reciprocal interaction between p53 and AKT in cisplatin-treated A549 cells, where p53 inhibits AKT, while AKT activates p53. In the case of cisplatin + JNK inhibitor SP600125, DNA damage and reactive oxygen species (ROS) contribute to cell death regulation in different ways. ROS exert opposite roles on cell fate-determining molecules p53 and AKT, and ROS act on p53 and AKT in opposite directions at low vs. high concentrations of cisplatin, combined or not with JNK inhibition. The differentially activated p53 in response to ROS (at low versus high concentrations of cisplatin, combined with JNK inhibitor) may be a molecular switch in the role of JNK from antiapoptotic to neutral/proapoptotic, and an executor of cell death. ROS is a possible threshold regulator that, together with an as-yet-unidentified factor, can differentially regulate p53. As a result, AKT phosphorylation and function are altered. The findings emphasize the importance of assessing the role of drug concentration when combining them with JNK inhibition when monitoring therapeutic efficacy and toxicity issues in personalized cancer treatment. Full article

(This article belongs to the Special Issue Structural Insights into Protein Kinases-Targeted Therapies: Overcoming Resistance and Advancing Precision Oncology)

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12 pages, 963 KB

Open AccessArticle

How Astragalin Modulates Glucose Uptake and Insulin Secretion in β-Cell Lines

by Paola Miranda Sulis, Alice Lima Rosa Mendes, Paula Waiss Zanusso Bunick, Karina Cesca, Carine Royer, Bruna Antunes Zaniboni, Fernanda Carvalho Cavalari, Guilherme Brasil Pintarelli, André Luiz Andreotti Dagostin and Fátima Regina Mena Barreto Silva

Pharmaceuticals 2026, 19(3), 508; https://doi.org/10.3390/ph19030508 - 20 Mar 2026

Viewed by 642

Abstract

Background/Objectives: Type 2 diabetes mellitus (T2DM) is characterized by chronic hyperglycemia and insulin resistance, leading to progressive metabolic dysfunction. Flavonoids, such as astragalin, have reported antidiabetic potential; however, their direct effects on pancreatic β-cell ionic mechanisms and insulin secretion remain unclear. This [...] Read more.

Background/Objectives: Type 2 diabetes mellitus (T2DM) is characterized by chronic hyperglycemia and insulin resistance, leading to progressive metabolic dysfunction. Flavonoids, such as astragalin, have reported antidiabetic potential; however, their direct effects on pancreatic β-cell ionic mechanisms and insulin secretion remain unclear. This study aimed to investigate the effects of astragalin on glucose uptake, insulin secretion, and membrane ionic currents in pancreatic β-cell lines. Methods: Murine MIN6 and rat INS-1 pancreatic β-cells were used as experimental models. Following astragalin treatment, glucose uptake was quantified by bioluminescence, and insulin secretion was measured by ELISA. Ionic currents were analyzed using the whole-cell patch-clamp technique. Selective pharmacological blockers targeting ATP-sensitive K⁺ channels (KATP), voltage-dependent K⁺ channels (K_v), and L-type voltage-dependent Ca²⁺ channels were applied to elucidate the underlying mechanisms. Results: Astragalin increased glucose uptake in a time-dependent manner, reaching a plateau between 3 and 5 h. Insulin secretion was significantly enhanced after 1 h of exposure to 100 µM astragalin. Patch-clamp recordings demonstrated that astragalin reduced potassium channel currents in pancreatic β-cells. Pharmacological modulation confirmed the involvement of KATP, K_v, and L-type Ca²⁺ channels. Verapamil attenuated the insulinotropic effect, supporting the role of calcium influx in astragalin-induced insulin exocytosis. Conclusions: Astragalin enhances glucose uptake and stimulates insulin secretion in pancreatic β-cells through modulation of potassium and calcium channels, promoting calcium-dependent exocytosis. These findings support its potential as a candidate for antidiabetic therapeutic strategies. Full article

(This article belongs to the Special Issue Natural Products in Diabetes Mellitus: 3rd Edition)

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19 pages, 612 KB

Open AccessReview

Hypersensitivity Reactions to Benzodiazepines Used for Perioperative Premedication: A Narrative Review

by Julia Gąsiorowska, Emilia Kiełczyńska, Weronika Dziamara, Amelia Grundys, Justyna Drozdowska, Monika Woźny, Aleksandra Skiba and Krzysztof Gomułka

Pharmaceuticals 2026, 19(3), 507; https://doi.org/10.3390/ph19030507 - 20 Mar 2026

Viewed by 1400

Abstract

Background: Hypersensitivity reactions to benzodiazepines, although uncommon, represent a clinically relevant issue in perioperative practice. Benzodiazepines are widely used medications with anxiolytic, sedative, and anticonvulsant properties. The objective of this review is to synthesize current knowledge on benzodiazepine hypersensitivity, focusing on underlying [...] Read more.

Background: Hypersensitivity reactions to benzodiazepines, although uncommon, represent a clinically relevant issue in perioperative practice. Benzodiazepines are widely used medications with anxiolytic, sedative, and anticonvulsant properties. The objective of this review is to synthesize current knowledge on benzodiazepine hypersensitivity, focusing on underlying mechanisms, clinical manifestations, diagnostic considerations, and strategies for management and prevention in the perioperative setting. Methodology: A narrative synthesis of the current literature on hypersensitivity reactions to benzodiazepines was performed. The review included published case reports, case series, and clinical studies describing hypersensitivity reactions, their clinical presentation, and diagnostic approaches. Particular attention was given to both immunological and non-immunological mechanisms, reported clinical phenotypes, and issues relevant to perioperative patient safety. Results: The extant evidence suggests that benzodiazepine hypersensitivity may involve both immunological and non-immunological pathways. The spectrum of reported reactions encompasses mild cutaneous manifestations and severe systemic responses, although the incidence remains low. This review highlights diagnostic challenges related to variable clinical presentation and the limited availability of standardized testing methods. Conclusions: Although cases of benzodiazepine hypersensitivity are uncommon, awareness of potential reactions is critical for ensuring safe clinical practice. This review emphasizes the necessity for additional research to elucidate the underlying mechanisms, standardize the diagnostic criteria, and formulate management protocols. Full article

(This article belongs to the Special Issue Advances in Drug Hypersensitivity Research)

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22 pages, 5409 KB

Open AccessArticle

Tailored Phytochitosomes as Targeted Nanotherapy for Alveolar Bone Regeneration in Diabetic Obese Rats

by Yosra S. R. Elnaggar, Mariam Zewail, Eman M. Salem, Wafaa Y. Alghonemy, Nevien M. Ahmed, Rania A. Hanafy, Waiel Daghistan, Ali M. Alaseem, Dina Khodeer, Elsayed G. Zaki, Ahmad N. Almougy and Mona A. Moustafa

Pharmaceuticals 2026, 19(3), 506; https://doi.org/10.3390/ph19030506 - 19 Mar 2026

Viewed by 618

Abstract

Background/Objectives: Individuals with diabetes often experience difficulties in the healing of their alveolar sockets. Furthermore, obesity is strongly associated with the development and progression of type 2 diabetes through complex metabolic and inflammatory mechanisms. The current study provides new insights into the [...] Read more.

Background/Objectives: Individuals with diabetes often experience difficulties in the healing of their alveolar sockets. Furthermore, obesity is strongly associated with the development and progression of type 2 diabetes through complex metabolic and inflammatory mechanisms. The current study provides new insights into the use of Luteolin (LU) and/or chitosan vesicles (CHV) to accelerate bone regeneration, highlighting a biologically and clinically relevant approach that leverages implants as a clinical solution. Methods: Sixty rats were randomly categorized into five groups: Group I (negative control); Group II (positive control), diabetic and obese rats; Group III (LU-treated), diabetic and obese rats with an extraction socket loaded with LU; Group IV (CHV-treated), diabetic and obese rats with an extraction socket loaded with CHV; and Group V (LU-CHV), diabetic and obese rats with an extraction socket loaded with LU-CHV. After 2 and 6 weeks, rats’ mandibles underwent histological, histomorphometric, biochemical, and statistical analyses. Results: The results demonstrated significant differences among the experimental groups. The LU-CHV formulation showed superior therapeutic performance compared with free luteolin and the untreated control group. In vitro release studies revealed sustained, controlled release from LU-CHV, whereas free luteolin exhibited rapid drug release. Additionally, LU-CHV significantly enhanced biological activity, as evidenced by improved anti-inflammatory and/or therapeutic markers compared to the other groups. These findings indicate that encapsulation within chitosan vesicles improved drug stability, bioavailability, and overall therapeutic efficiency. Conclusions: LU-CHV demonstrated superior efficacy compared to free luteolin, highlighting the advantage of chitosan-based vesicular delivery systems. LU-CHV not only enhanced controlled drug release and therapeutic outcomes but also presents a promising platform that could significantly advance targeted drug delivery strategies in inflammatory and metabolic disorders. The findings suggest that LU-CHV represents a transformative approach in improving treatment effectiveness and patient outcomes. Full article

(This article belongs to the Special Issue Drugs and Implants in Orthopedic Surgery and Traumatology)

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13 pages, 3228 KB

Open AccessArticle

Employment of a Resazurin Viability-Based Assay for Minimum Inhibitory and Bactericidal Concentration Determination

by Lorena G. Calvo, Stephanya Corral-Orbe, Rosa-Antía Villarino, Sandra Sánchez and Trinidad de Miguel

Pharmaceuticals 2026, 19(3), 505; https://doi.org/10.3390/ph19030505 - 19 Mar 2026

Cited by 1 | Viewed by 1271

Abstract

Background/Objectives: The increasing prevalence of antimicrobial-resistant bacteria highlights the need for improved methodologies to evaluate antimicrobial activity beyond conventional minimum inhibitory concentration testing. While resazurin-based assays are widely used for minimum inhibitory concentration determination due to their simplicity and sensitivity, minimum bactericidal [...] Read more.

Background/Objectives: The increasing prevalence of antimicrobial-resistant bacteria highlights the need for improved methodologies to evaluate antimicrobial activity beyond conventional minimum inhibitory concentration testing. While resazurin-based assays are widely used for minimum inhibitory concentration determination due to their simplicity and sensitivity, minimum bactericidal concentration assessment still relies on labor-intensive colony-forming unit counting. The objective of this study was to develop and validate a resazurin-based microwell assay capable of determining both the minimum inhibitory concentration and the minimum bactericidal concentration without routine plate counting, thereby simplifying bactericidal evaluation. Methods: A two-step resazurin-based fluorescence assay was designed and performed in microplates. After determining the minimum inhibitory concentration using resazurin as a metabolic indicator, well-showing inhibited bacterial growths were subjected to a regrowth phase by transferring aliquots into fresh antimicrobial-free medium containing resazurin. This additional step allowed discrimination between reversible metabolic inhibition and irreversible bacterial death. The method was evaluated using ciprofloxacin and chloramphenicol against four bacterial species: Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. Minimum bactericidal concentration values obtained using this assay were compared with those obtained through conventional colony counting on agar plates. Results: Minimum bactericidal concentration values obtained using the two-step fluorescence assay were fully concordant with the conventional colony-forming unit counting method for all tested antibiotics and bacterial species. Conclusions: The proposed two-step resazurin-based microwell assay represents a rapid, reliable, and less labor-intensive alternative for the determination of both the minimum inhibitory concentration and the minimum bactericidal concentration, with potential applications in clinical and industrial microbiology laboratories. Full article

(This article belongs to the Section Natural Products)

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18 pages, 12080 KB

Open AccessArticle

Ivabradine Attenuates Experimental Hepatic Fibrosis by Modulating Inflammatory and Apoptotic Signaling Pathways

by Salman H. Alotaibi, Mahmoud M. Samaha, Manar G. Helal and Dina S. El-Agamy

Pharmaceuticals 2026, 19(3), 504; https://doi.org/10.3390/ph19030504 - 19 Mar 2026

Viewed by 649

Abstract

Background: Hepatic fibrosis and its progressive form, liver cirrhosis, are dangerously recognized complications of liver injury with limited treatment options. This study evaluated the hepatoprotective effects of ivabradine on thioacetamide (TAA)-induced hepatic fibrosis in rats. Methods: Rats were divided into five [...] Read more.

Background: Hepatic fibrosis and its progressive form, liver cirrhosis, are dangerously recognized complications of liver injury with limited treatment options. This study evaluated the hepatoprotective effects of ivabradine on thioacetamide (TAA)-induced hepatic fibrosis in rats. Methods: Rats were divided into five groups with 10 rats/group and treated as follows: normal, where rats received 0.5% CMC-Na solution orally; ivabradine control, where rats received only ivabradine (20 mg/kg, once daily, orally) for 6 weeks; TAA, where rats received an intraperitoneal (i.p.) injection of TAA (200 mg/kg) thrice weekly for 6 weeks and daily oral 0.5% CMC-Na solution, and two ivabradine + TAA groups, where two doses of ivabradine were tested. Low (10 mg/kg) and high (20 mg/kg) doses of ivabradine were orally given once daily to each group for 6 weeks concurrently with TAA injection. Results: TAA caused marked elevations in liver enzymes, increased MDA, depletion of antioxidant defenses, activation of NF-κB p65 and pro-inflammatory cytokines, dysregulation of apoptotic markers, and upregulation of the PI3K/AKT/mTOR and TGF-β pathways, accompanied by extensive collagen deposition. Ivabradine produced dose-dependent improvements in biochemical markers of liver function, restored oxidant/antioxidant balance, suppressed NF-κB p65/TNF-α, normalized Bax/Bcl-2/caspase-3 expression, and inhibited PI3K/AKT/mTOR as well as TGF-β signaling, leading to significant attenuation of fibrosis. Conclusions: The current findings indicate that ivabradine exerts potent antioxidant, anti-inflammatory, anti-apoptotic, and antifibrotic actions against TAA-induced hepatic fibrosis. Future clinical studies are recommended to determine whether these protective effects translate to patients with chronic liver disease. Full article

(This article belongs to the Section Pharmacology)

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28 pages, 4809 KB

Open AccessArticle

Exploring the Multifaceted Phytochemical Profile of Nigella sativa and the Therapeutic Potential of Thymoquinone

by Mohamed A. Fareid, Gamal M. El-Sherbiny, Nancy M. Elafandy, Nagat E. Eltoum, Mohamed S. Othman, Mohamed Shawky, Ahmad S. El-Hawary, Fatma A. Hamada and Amira Salah El-Din Youssef

Pharmaceuticals 2026, 19(3), 503; https://doi.org/10.3390/ph19030503 - 18 Mar 2026

Viewed by 1220

Abstract

Background: Nigella sativa (black cumin) seeds are renowned for their ethnomedicinal significance and are rich in bioactive phytochemicals, which contribute to food preservation and the prevention of various diseases through their antimicrobial and antioxidant properties. Accordingly, this study aimed to characterize the [...] Read more.

Background: Nigella sativa (black cumin) seeds are renowned for their ethnomedicinal significance and are rich in bioactive phytochemicals, which contribute to food preservation and the prevention of various diseases through their antimicrobial and antioxidant properties. Accordingly, this study aimed to characterize the phytochemical composition of N. sativa seed extracts, isolate thymoquinone, and assess their antibacterial, antibiofilm, antioxidant, anti-inflammatory and antidiabetic activities. Methods: Nigella sativa seed extracts were prepared using solvents of increasing polarity and analyzed for phytochemical content. Metabolite profiling was performed using UHPLC/QTOF-MS. Thymoquinone, the major constituent, was isolated via thin-layer chromatography (TLC), further purified using semi-preparative reverse-phase high-performance liquid chromatography (RP-HPLC), and evaluated in vitro for antibacterial, antibiofilm, antioxidant, anti-inflammatory, and antidiabetic activities. Results: Extraction yields ranged from 5.5% to 8.4% (w/w), with methanol yielding the highest phenol (6.34 ± 0.31 mg GAE/mL) and flavonoid (5.12 ± 0.26 mg QE/mL) contents. UHPLC/QTOF-MS revealed a chemically diverse profile dominated by thymoquinone (58% relative abundance), alongside p-cymene, carvacrol, longifolene, and nigellidine. Thymoquinone (Rf = 0.56) was initially isolated from the methanolic extract with a yield of 270 mg/g and further purified from preparative TLC fractions using semi-preparative RP-HPLC, affording 82 mg of >95% pure compound with a 68.3% recovery, suitable for subsequent biological assays. It inhibited Gram-positive and Gram-negative bacteria, with MICs of 62.5 µg/mL against Staphylococcus aureus, Bacillus subtilis, and Listeria monocytogenes; 125–250 µg/mL against Escherichia coli and Salmonella typhimurium; and 500 µg/mL against Pseudomonas aeruginosa. Thymoquinone reduced biofilm formation (>80% at 25–50 µg/mL; MBIC₅₀ ≈ 5.4–11.6 µg/mL), exhibited antioxidant activity (DPPH IC₅₀ = 52.3 ± 2. 1 µg/mL; ABTS IC₅₀ = 41.6 ± 1.9 µg/mL), stabilized erythrocyte membranes (IC₅₀ ≈ 14.8 µg/mL), and inhibited carbohydrate-hydrolyzing enzymes, with stronger inhibition of α-glucosidase (~92%) than α-amylase (~84%) at 128 µg/mL. Conclusions: Thymoquinone is a major bioactive constituent of N. sativa seeds, exhibiting consistent multi-target in vitro activity. These findings highlight its functional relevance and in vivo investigations to establish therapeutic potential. Full article

(This article belongs to the Special Issue Medicinal Plants: A Natural Approach to Inflammatory Diseases and Conditions Related to Oxidative Stress)

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40 pages, 17948 KB

Open AccessArticle

Yixin Yangshen Granules Target HIF−1 Signaling to Modulate the Neuroimmune Microenvironment in Alzheimer’s Disease: Insights from Integrative Multi-Omics and Deep Learning

by Zhihao Wang, Linshuang Wang, Yusheng Zhang, Sixia Yang, Bo Shi, Dasheng Liu, Han Zhang, Wan Xiao, Junying Zhang, Xuejie Han and Dongfeng Wei

Pharmaceuticals 2026, 19(3), 502; https://doi.org/10.3390/ph19030502 - 18 Mar 2026

Viewed by 835

Abstract

Background/Objectives: Alzheimer’s disease (AD) involves amyloid and tau pathology with neuroimmune dysregulation, and Yixin Yangshen Granules (YXYS) shows neuroprotective promise, though mechanisms remain unclear. This study aimed to elucidate the multi-target mechanisms of YXYS in AD. Methods: The study began by [...] Read more.

Background/Objectives: Alzheimer’s disease (AD) involves amyloid and tau pathology with neuroimmune dysregulation, and Yixin Yangshen Granules (YXYS) shows neuroprotective promise, though mechanisms remain unclear. This study aimed to elucidate the multi-target mechanisms of YXYS in AD. Methods: The study began by analyzing a public human AD hippocampal snRNA-seq dataset to identify cell-type-specific pathological pathways and profiled YXYS constituents by UPLC-QTOF-MS. In vitro, YXYS cytoprotection against mitochondrial dysfunction and oxidative stress was tested in Aβ_25–35-challenged HT22 cells; in vivo efficacy was assessed in Aβ_1–42-induced mice via behavioral and histopathological analyses. Integrated transcriptomic and proteomic profiling of brain tissue, with ELISA, qRT-PCR, and Western blot validation, confirmed pathway targets. Using the intersection of transcriptomic and proteomic targets as biological input, the DTIAM deep learning framework was employed to prioritize active YXYS constituents. Finally, molecular docking and 100-ns dynamics simulations demonstrated direct binding of Ganosporelactone A to HIF−1α. Results: AD snRNA-seq analysis highlighted HIF−1 and AGE-RAGE signaling as prominent pathways in the AD hippocampus, particularly enriched in brain microvascular endothelial cells, implicating neurovascular hypoxic and inflammatory stress. In Aβ-induced mice, YXYS improved cognition, reduced Aβ pathology, suppressed neuroinflammation, and promoted neuronal survival, consistent with in vitro evidence of restored mitochondrial function. Multi-omics confirmed convergence on HIF−1 and AGE-RAGE pathways, with YXYS rebalancing the neuroimmune microenvironment by reducing pro-inflammatory M0 macrophages. Screening against these consensus signaling hubs, deep learning analysis prioritized Ganosporelactone A as the top-ranked modulator, and molecular further demonstrated the stable binding of Ganosporelactone A to HIF−1α, linking YXYS to mitigation of hypoxic stress. Conclusions: Guided by multi-omics and deep learning, our findings suggest that YXYS may alleviate AD-related phenotypes through multi-target modulation of the HIF−1 and AGE-RAGE pathways, with associated improvements in neuro-immune homeostasis and reductions in oxidative stress, neuroinflammation, and hypoxia. Full article

(This article belongs to the Special Issue The Role of Phytochemicals in Aging and Aging-Related Diseases, 2nd Edition)

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10 pages, 841 KB

Open AccessArticle

Radioimmunotherapy for Malignant Mesothelioma Targeting C-ERC/Mesothelin

by Hirofumi Hanaoka, Aiko Yamaguchi, Masahiro Maeda, Tatsuya Segawa and Noboru Oriuchi

Pharmaceuticals 2026, 19(3), 501; https://doi.org/10.3390/ph19030501 - 18 Mar 2026

Viewed by 469

Abstract

Background/Objectives: Malignant mesothelioma has a poor prognosis and limited therapeutic options. C-ERC/mesothelin is highly expressed in mesotheliomas and is a potential target for radioimmunotherapy (RIT). This study evaluated the radiolabeled anti-C-ERC/mesothelin antibody mAb 22A31 as a therapeutic agent. Methods: C-ERC/mesothelin expression [...] Read more.

Background/Objectives: Malignant mesothelioma has a poor prognosis and limited therapeutic options. C-ERC/mesothelin is highly expressed in mesotheliomas and is a potential target for radioimmunotherapy (RIT). This study evaluated the radiolabeled anti-C-ERC/mesothelin antibody mAb 22A31 as a therapeutic agent. Methods: C-ERC/mesothelin expression in mesothelioma cell lines was assessed by Western blotting, and the specific binding of ¹²⁵I-labeled mAb 22A31 was examined. Biodistribution of ¹¹¹In-labeled mAb 22A31 was evaluated in a mesothelioma cell line, MSTO-211H tumor-bearing mice. The therapeutic efficacy of ⁹⁰Y-labeled mAb 22A31 was evaluated in subcutaneous and pleural dissemination models. Results: mAb 22A31 showed specific binding considering the level of C-ERC/mesothelin expression in each mesothelioma cell line. ¹¹¹In-mAb 22A31 accumulated in tumors with minimal uptake in normal tissues. ⁹⁰Y-mAb 22A31 significantly delayed the growth of subcutaneous tumors and improved survival in a pleural dissemination model. Conclusions: Radiolabeled mAb 22A31 specifically targeted C-ERC/mesothelin and demonstrated therapeutic efficacy in a mesothelioma xerograph model. Therefore, ⁹⁰Y-mAb 22A31 is a promising RIT agent and supports the further development of C-ERC/mesothelin-targeted therapy for mesothelioma. Full article

(This article belongs to the Special Issue Advances in Antibody–Drug Conjugates)

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23 pages, 5199 KB

Open AccessArticle

Biluo Qianyuan Formula Ameliorates Post-Traumatic Osteoarthritis by Suppressing FN1-Mediated Synovial Inflammation and Restoring Joint Homeostasis

by Yinqiu Wu, Guangran Hu, Shengzhe Zhang, Guilan Jin and Hua Dai

Pharmaceuticals 2026, 19(3), 500; https://doi.org/10.3390/ph19030500 - 18 Mar 2026

Viewed by 573

Abstract

Background: Post-traumatic osteoarthritis (PTOA) lacks effective disease-modifying therapies that preserve joint structure while promoting tissue repair. This study aimed to evaluate the therapeutic efficacy and underlying mechanism of Biluo Qianyuan Formula (BLQYF), a standardized herbal formulation derived from clinical practice, as a [...] Read more.

Background: Post-traumatic osteoarthritis (PTOA) lacks effective disease-modifying therapies that preserve joint structure while promoting tissue repair. This study aimed to evaluate the therapeutic efficacy and underlying mechanism of Biluo Qianyuan Formula (BLQYF), a standardized herbal formulation derived from clinical practice, as a potential disease-modifying alternative to celecoxib in a murine model of PTOA. Methods: A murine PTOA model was established and treated with BLQYF at different doses, with celecoxib serving as a pharmacological comparator. Safety was assessed by hepatic and renal toxicity analyses. Therapeutic effects were evaluated using micro-computed tomography (micro-CT) and histological staining. Network-based integrative analyses were conducted to identify key regulatory targets, followed by experimental validation in fibroblast-like synoviocytes. Results: BLQYF was well tolerated under the experimental conditions, with no detectable hepatic or renal toxicity at therapeutic doses. Micro-CT and histological analyses demonstrated that BLQYF dose-dependently mitigated subchondral bone deterioration, enhanced cartilage regeneration, and restored collagen deposition. At higher doses, BLQYF showed therapeutic efficacy comparable to celecoxib, with superior outcomes regarding cartilage reparation. Mechanistically, integrative analyses identified fibronectin 1 (FN1) as a central regulatory hub. Validation experiments confirmed that BLQYF suppressed FN1, MMP3, and TGF-β expression in fibroblast-like synoviocytes, thereby attenuating inflammation and extracellular matrix degradation. Conclusions: These findings support BLQYF as a promising disease-modifying therapeutic candidate for PTOA and highlight the fibroblast–FN1 axis as a novel pharmacological target for intervention. Full article

(This article belongs to the Section Pharmacology)

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18 pages, 24132 KB

Open AccessArticle

Myricetin Inhibits Osteosarcoma Cell Viability and Modulates EMT-Related Genes Associated with the SNAI1/MMP-9 Axis

by Isabela Santos, Hélio M. T. Albuquerque, Marta Teixeira Pinto, Nuno Mendes, José Miguel P. Ferreira de Oliveira and Eduarda Fernandes

Pharmaceuticals 2026, 19(3), 499; https://doi.org/10.3390/ph19030499 - 18 Mar 2026

Viewed by 572

Abstract

Background/Objectives: Osteosarcoma treatment options remain limited due to tumor metastasis and the toxicity of conventional chemotherapy, warranting new therapeutic strategies. A well-founded strategy is the use of flavonoids, a class of phytochemicals possessing pharmaceutical properties that contribute to anticancer effects, including antioxidant and [...] Read more.

Background/Objectives: Osteosarcoma treatment options remain limited due to tumor metastasis and the toxicity of conventional chemotherapy, warranting new therapeutic strategies. A well-founded strategy is the use of flavonoids, a class of phytochemicals possessing pharmaceutical properties that contribute to anticancer effects, including antioxidant and anti-inflammatory properties. This study aimed to evaluate the anticancer potential of flavonoids in osteosarcoma and investigate their interaction with doxorubicin. Methods: In this study, five flavonoids were screened for cytotoxicity and selectivity across four osteosarcoma cell lines and healthy fibroblasts (MRC-5). The interaction between myricetin and doxorubicin was assessed using a fixed-ratio combination approach. Cell migration and invasion were evaluated using cell exclusion/wound healing and 2D co-culture assays. EMT-related gene expressions were assessed by RT-qPCR. Antitumor activity was evaluated in vivo using a chick chorioallantoic membrane (CAM) xenograft model. Results: Myricetin emerged as the most selective compound, exhibiting cytotoxicity against osteosarcoma cells while sparing MRC-5 fibroblasts. Notably, myricetin synergized with doxorubicin (ratio 69:1), enhancing its cytotoxicity and significantly reducing osteosarcoma cell migration in vitro. Myricetin downregulated SNAI1 and MMP9, suggesting modulation of epithelial–mesenchymal transition (EMT)-related pathways. Complementarily, in the CAM xenograft model, myricetin reduced xenograft tumor size, confirming its anticancer activity in vivo. Conclusions: Collectively, these findings emphasize the anticancer potential of myricetin in osteosarcoma through inhibition of the SNAI1/MMP-9 signaling axis. Full article

(This article belongs to the Special Issue Pharmacological Activities of Flavonoids and Their Analogues, Third Edition)

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21 pages, 10490 KB

Open AccessArticle

A Data-Driven Approach for Interpretable and Efficient Predictive Modeling: A Case Study in SARS-CoV-2 Protease Inhibitor Discovery Through Feature Selection

by Branislav Stanković, Sang-Yong Oh and Dušan Ramljak

Pharmaceuticals 2026, 19(3), 498; https://doi.org/10.3390/ph19030498 - 18 Mar 2026

Viewed by 457

Abstract

Background/Objectives: Feature selection approaches should satisfy all evaluation criteria required by state-of-the-art chemoinformatic models. Our aim is to develop a methodology that is robust, interpretable and computationally efficient. Methods: This study presents a robust methodology for developing highly interpretable and computationally [...] Read more.

Background/Objectives: Feature selection approaches should satisfy all evaluation criteria required by state-of-the-art chemoinformatic models. Our aim is to develop a methodology that is robust, interpretable and computationally efficient. Methods: This study presents a robust methodology for developing highly interpretable and computationally efficient predictive models, with a specific application in the discovery of SARS-CoV-2 main protease inhibitors. We evaluated various descriptor selection procedures to identify a transparent and reproducible approach that provides actionable insights for data-driven decisions. The models were trained and tested using molecules from the CHEMBL database and further validated on an external set of compounds. Results: Our findings demonstrate that a recently proposed procedure, combining the FeatureWiz algorithm with stepwise feature selection, is the only approach that satisfies all evaluation criteria required by state-of-the-art chemoinformatic models. In particular, we found that models based on two-dimensional descriptors and Ordinary Least Squares regression achieved the best results. Conclusions: Our framework and the choices made offer significant advantages in a decision-making context due to their inherent interpretability and computational efficiency. Our derived models, benchmarked against those in the literature, serve as effective, transparent tools for the rapid and reliable prediction of biological activity, providing a validated framework for data-driven decisions in drug discovery and beyond. Full article

(This article belongs to the Section Medicinal Chemistry)

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39 pages, 5739 KB

Open AccessReview

NRF2 as a Therapeutic Target in Dermatological Disorders: Mechanisms and Molecules

by Ismael Khiar-Fernández, Nora Khiar-Fernández, José-Juan Pereyra-Rodríguez and Inmaculada Fernández

Pharmaceuticals 2026, 19(3), 497; https://doi.org/10.3390/ph19030497 - 17 Mar 2026

Cited by 1 | Viewed by 1210

Abstract

The nuclear factor erythroid 2–related factor 2 (NRF2) is a master transcription factor that orchestrates cellular defense against oxidative and electrophilic stress. Dysregulation of the KEAP1–NRF2–ARE pathway has been implicated in several dermatological disorders, including vitiligo, psoriasis, atopic dermatitis, photoaging, and radiation dermatitis. [...] Read more.

The nuclear factor erythroid 2–related factor 2 (NRF2) is a master transcription factor that orchestrates cellular defense against oxidative and electrophilic stress. Dysregulation of the KEAP1–NRF2–ARE pathway has been implicated in several dermatological disorders, including vitiligo, psoriasis, atopic dermatitis, photoaging, and radiation dermatitis. This review summarizes recent advances in the understanding of NRF2 activation mechanisms and highlights pharmacological and natural compounds with potential dermatological applications. A comprehensive analysis of natural, semisynthetic, and synthetic NRF2 modulators is provided, describing their chemical structures, synthetic approaches, mechanisms of action, preclinical and clinical evidence, and therapeutic relevance for skin disorders. Multiple classes of NRF2 activators, including isothiocyanates such as sulforaphane, triterpenoids such as omaveloxolone, flavonoids including baicalein and apigenin, alkaloids such as berberine, glycosides like afzelin and paeoniflorin, stilbenoids such as tapinarof, and α,β-unsaturated fumaric acid esters such as dimethyl fumarate, have demonstrated antioxidant, anti-inflammatory, and cytoprotective effects in keratinocytes and melanocytes. Some of these agents, particularly dimethyl fumarate and tapinarof, have advanced to clinical development or commercialization, whereas others remain at the preclinical stage but show encouraging results in animal models and cell culture systems. Overall, pharmacological activation of NRF2 represents a promising therapeutic strategy to counteract oxidative stress–driven skin damage and inflammation; however, continued translational and clinical research is required to optimize formulations, dosing regimens, and safety profiles for integration into dermatological practice. Full article

(This article belongs to the Collection Feature Review Collection in Medicinal Chemistry)

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23 pages, 3923 KB

Open AccessArticle

Curcumin in High Doses Reverses the UV-B-Induced DNMT and HDAC Upregulation In Vitro: A Novel Anti-Cancer Approach?

by Afshin Zand, Bence L. Raposa, Dávid Szép, John M. Macharia, Ghodratollah Nowrasteh, Ferenc Budán and Tímea Varjas

Pharmaceuticals 2026, 19(3), 496; https://doi.org/10.3390/ph19030496 - 17 Mar 2026

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Abstract

Background/Objectives: The primary mechanisms driving UV-induced carcinogenesis include DNA damage leading to mutations, and reactive oxygen species (ROS) formation that can cause inflammation, immunosuppression, alteration of the structure of proteins, including transcription factors, and carcinogenesis through epigenetic modifications. Curcumin has the potential to [...] Read more.

Background/Objectives: The primary mechanisms driving UV-induced carcinogenesis include DNA damage leading to mutations, and reactive oxygen species (ROS) formation that can cause inflammation, immunosuppression, alteration of the structure of proteins, including transcription factors, and carcinogenesis through epigenetic modifications. Curcumin has the potential to inhibit DNA-methyltransferases (DNMTs) and histone deacetylases (HDACs), but this has not been examined yet at the gene-expression level. In this article, we aimed to explore the potential protective effect of curcumin against UV radiation-induced DNMT1, DNMT3A, DNMT3B, HDAC5, and HDAC6 expression in immortalized keratinocytes (HaCaT), hepatocellular carcinoma (HepG2), and lung adenocarcinoma (A549) cells. Methods: Cells were exposed to UV-B radiation for different periods and treated with curcumin at different concentrations to evaluate dose-related trends in DNMT and HDAC gene expression compared with untreated UV-exposed cells. Results: UV exposure increased the DNMT and HDAC gene expression levels in the examined cells dose-dependently. Curcumin exposure resulted in decreased mRNA expression levels of DNMT and HDAC gene expression. In our experimental setup curcumin modulated the transcription of DNMT and HDAC genes in A549 and HaCaT cells in a dose-dependent manner. In HepG2 cells, UV-B induced a less pronounced, but still significant, increase in the examined gene expression levels. This effect was also dose-dependently decreased by curcumin, although less markedly. Conclusions: Future studies are warranted to examine if curcumin combined with other chemopreventive agents through the HDAC and DNMT inhibitory activity at the gene expression level can exert a synergistic effect and may potentially supplement cancer therapeutic strategies. Full article

(This article belongs to the Special Issue Targeting Epigenetic Regulation for Cancer Therapy)

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25 pages, 12954 KB

Open AccessArticle

From a Multi-Omics Signature to a Therapeutic Candidate: Computational Prediction and Experimental Validation in Liver Fibrosis

by Yingying Qin, Shuoshuo Ma, Haoyuan Hong, Deyuan Zhong, Yuxin Liang, Yuhao Su, Yahui Chen, Xing Chen, Yizhun Zhu and Xiaolun Huang

Pharmaceuticals 2026, 19(3), 495; https://doi.org/10.3390/ph19030495 - 17 Mar 2026

Viewed by 1133

Abstract

Background: Advanced liver fibrosis (LF) is a major determinant of prognosis across chronic liver diseases. Current biomarkers are often etiology-specific and lack cross-cohort robustness. Shared molecular drivers across etiologies remain incompletely defined, and effective anti-fibrotic therapies are limited. Methods: We developed [...] Read more.

Background: Advanced liver fibrosis (LF) is a major determinant of prognosis across chronic liver diseases. Current biomarkers are often etiology-specific and lack cross-cohort robustness. Shared molecular drivers across etiologies remain incompletely defined, and effective anti-fibrotic therapies are limited. Methods: We developed a multi-algorithm consensus machine-learning framework to derive a robust LF progression signature. In the training non-alcoholic fatty liver disease (NAFLD) cohort GSE213621 (n = 368), samples were formulated as a binary classification task (mild fibrosis, F0–F2; advanced fibrosis, F3–F4). Candidate genes were screened in parallel using Boruta, Least Absolute Shrinkage and Selection Operator (LASSO), random forest, and eXtreme Gradient Boosting (XGBoost). Genes selected by at least two algorithms were defined as a high-consensus pool, and genes consistently selected by all four algorithms were prioritized to construct a core signature. Model performance was evaluated by stratified cross-validation in the training cohort and externally validated in four independent cohorts of different etiologies (GSE49541, GSE84044, GSE130970, and GSE276114). Cellular sources of signature genes were characterized using single-cell RNA sequencing (scRNA-seq) datasets GSE136103 (human) and GSE172492 (mouse). For therapeutic discovery, the high-consensus expression profile was queried against the Connectivity Map (CMap) to prioritize compounds predicted to reverse the fibrotic transcriptional program. Withaferin A (WFA) was selected for experimental validation in a carbon tetrachloride (CCl₄)-induced mouse LF model and in the transforming growth factor-β1 (TGF-β1)-stimulated human hepatic stellate cell line LX-2. Bulk liver RNA-seq profiling was performed to interrogate WFA-associated molecular changes in vivo. Results: We identified a six-gene signature (CLEC4M, COL25A1, ITGBL1, NALCN, PAPPA, and PEG3) that discriminated advanced from mild fibrosis, achieving a mean AUC of 0.890 in internal cross-validation and an average AUC of 0.864 across external validation cohorts. scRNA-seq analysis revealed cell-type-specific expression with prominent enrichment in fibroblast populations. In vivo, WFA markedly attenuated CCl₄-induced fibrosis (p < 0.05) and reversed 1314 fibrosis-associated differentially expressed genes (adjusted p < 0.05), which were enriched in fatty acid metabolism and PPAR signaling, as well as extracellular matrix (ECM)–receptor interaction and focal adhesion (adjusted p < 0.05). In vitro, WFA suppressed TGF-β1-induced LX-2 activation, reducing α-SMA and Fibronectin expression (p < 0.05). Conclusions: We report a six-gene signature that robustly predicts advanced LF across etiologies, define its cellular context using single-cell atlases, and validate the anti-fibrotic activity of WFA in both in vivo and in vitro models. Bulk liver RNA-seq and cellular evidence further suggest that WFA-associated effects are linked to lipid metabolic programs, ECM remodeling, and attenuation of hepatic stellate cell activation. Full article

(This article belongs to the Section Medicinal Chemistry)

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16 pages, 5689 KB

Open AccessArticle

Potential Value of a Combination of Polypodium leucotomos and Aspalathus linearis Extracts in Protecting Vitamin D Receptor Levels During Skin Oxidative Stress

by Marta Mascaraque, María Gallego-Rentero, Andrea Barahona-López, Paula Cano, Ángeles Juarranz, Ana López Sánchez and Salvador González

Pharmaceuticals 2026, 19(3), 494; https://doi.org/10.3390/ph19030494 - 17 Mar 2026

Viewed by 762

Abstract

Background/Objectives: Vitamin D (VD), through the interaction with its receptor (VDR), plays essential roles in the skin. VDR-mediated signaling prevents cancer development and improves prognosis, making it an appealing target for therapy. However, VD cutaneous synthesis begins with solar exposure, which is the [...] Read more.

Background/Objectives: Vitamin D (VD), through the interaction with its receptor (VDR), plays essential roles in the skin. VDR-mediated signaling prevents cancer development and improves prognosis, making it an appealing target for therapy. However, VD cutaneous synthesis begins with solar exposure, which is the first etiological factor for cutaneous cancer and increases oxidative stress (OS). This complicates the dermatologist’s perspective when advising photoprotective strategies while aiming to consider the benefits of VD signaling. In this context, and in the absence of cutaneous data to date, this research aims to address VDR dynamics in skin cells and tissue subjected to OS. It also explores the potential of a natural photoprotectant with antioxidant properties (a specific combination of Polypodium leucotomos and Aspalathus linearis extracts) in preventing VDR depletion. Methods: HaCaT cell cultures and skin explants were used as experimental models. OS was induced by treatments with hydrogen peroxide (H₂O₂). The proteins of interest (VDR and Nuclear Factor Erythroid 2-Related Factor 2 (NRF2)) were analyzed by immunostaining. Cell viability, nuclear counterstaining, and Haematoxylin/Eosin staining were used as cyto/histochemical controls. Results: In both experimental models, we observed the reduction of VDR under OS. Pre-treatments with the botanical ingredient preserved VDR levels from that decline, probably through a mechanism involving NRF2. Conclusions: Cutaneous VDR levels are altered under oxidative stress, and certain photoprotectants could preserve them. This opens the door to preserving the benefits of VDR signaling while preventing solar radiation damage, bringing a new viewpoint for designing future strategies in skin cancer prevention and treatment. Full article

(This article belongs to the Special Issue Anticancer Compounds in Medicinal Plants—4th Edition)

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19 pages, 2580 KB

Open AccessArticle

Quantitative Analysis of the Vitamin D₃ Content in Dietary Supplements Marketed in Hungary Using High-Performance Liquid Chromatography

by András Nagy, Róbert György Vida, Eszter Fliszár-Nyúl, Gábor Lovász, Katalin Fábián and Gábor Pozsgai

Pharmaceuticals 2026, 19(3), 493; https://doi.org/10.3390/ph19030493 - 17 Mar 2026

Viewed by 1426

Abstract

Background/Objectives: The use of over-the-counter vitamin D₃ supplements has increased substantially in recent years. Compared with pharmaceuticals, dietary supplements are subject to less stringent regulatory oversight, raising concerns regarding labeling accuracy, consumer knowledge, and patient safety. This study aimed to assess public [...] Read more.

Background/Objectives: The use of over-the-counter vitamin D₃ supplements has increased substantially in recent years. Compared with pharmaceuticals, dietary supplements are subject to less stringent regulatory oversight, raising concerns regarding labeling accuracy, consumer knowledge, and patient safety. This study aimed to assess public knowledge and preferences related to vitamin D₃ supplementation and to evaluate the content accuracy and short-term stability of commonly used products. Methods: A cross-sectional online survey containing 39 questions was conducted in Hungary between 1 May and 30 June 2024. Based on survey responses, the most frequently used vitamin D₃ supplements (five soft gel capsules and four tablets) were selected for laboratory analysis. Vitamin D₃ content was quantified using a validated high-performance liquid chromatography (HPLC) method with UV detection. Soft gel capsules were additionally exposed to natural daylight for one month to assess short-term photostability. Results: In total, 367 participants (mean age 31.0 ± 12.5 years) completed the survey, and only 3.5% answered correctly all knowledge-based questions. Six commonly reported supplement brands accounted for approximately 90% of responses. Measured vitamin D₃ content remained within the tolerance limit (−20% to +50%). Following sunlight exposure, three of four capsule products showed no substantial vitamin D₃ loss, while one exhibited a 14.7% decrease. Conclusions: Most analyzed vitamin D₃ supplements complied with labeled content claims, but substantial knowledge gaps were identified that may affect patient safety. The validated HPLC method supports pharmacovigilance-oriented quality monitoring of vitamin D₃ supplements and underscores the need for improved professional counseling. Full article

(This article belongs to the Special Issue Pharmacovigilance Insights: Addressing Drug Misuse and Enhancing Drug Safety)

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15 pages, 2497 KB

Open AccessArticle

Berberine-Mediated BCRP Inhibition Enhances Systemic Exposure of Rhein: A Study to Unravel the Pharmacokinetic Basis of Synergy in Da-Huang-Xiao-Shi Decoction

by Zhangyao Xu, Hongyu Li, Haoyu Xue, Xiaoge Wang, Tianming Wang, Yuyang Zhou, Jifeng Gu and Rong Shi

Pharmaceuticals 2026, 19(3), 492; https://doi.org/10.3390/ph19030492 - 17 Mar 2026

Cited by 1 | Viewed by 617

Abstract

Background/Objectives: Cholestasis is a clinically intractable liver disorder. Da-Huang-Xiao-Shi Decoction (DHXSD), a classic traditional Chinese medicine formula, demonstrates notable efficacy, yet the mechanistic basis for its multi-herb synergy remains unclear. The purpose of this study was to decipher the pharmacokinetic interaction underlying [...] Read more.

Background/Objectives: Cholestasis is a clinically intractable liver disorder. Da-Huang-Xiao-Shi Decoction (DHXSD), a classic traditional Chinese medicine formula, demonstrates notable efficacy, yet the mechanistic basis for its multi-herb synergy remains unclear. The purpose of this study was to decipher the pharmacokinetic interaction underlying the synergy of DHXSD. Methods: A cholestatic rat model was established in male Sprague Dawley rats. Hepatoprotective efficacy was evaluated, and the pharmacokinetics of anthraquinones were profiled. Key interaction mechanisms were investigated using the everted intestinal sac model, the breast cancer resistance protein (BCRP)-overexpressing MDCKII cells, and molecular docking simulations. Results: DHXSD provided significantly stronger hepatoprotection than its principal herb Rheum palmatum L. (DaHuang, DH) alone. This enhanced efficacy correlated with an approximate 2-fold increase in the systemic exposure of rhein compared to DH monotherapy. We identified berberine from Phellodendron amurense Rupr. (Huang Bo, HB) as the key synergist, which potently inhibited the BCRP efflux transporter, thereby enhancing rhein absorption. In contrast, geniposide from Gardenia jasminoides Ellis (Zhi Zi, ZZ) showed minimal effects. Conclusions: This work elucidates a concrete, transporter-mediated pharmacokinetic interaction as the core mechanism underlying herbal synergy in DHXSD. Our findings offer a rational strategy—targeted efflux transporter modulation—for improving the oral bioavailability of challenging drug molecules. Full article

(This article belongs to the Section Natural Products)

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