Pharmaceuticals

12 pages, 425 KiB

Open AccessArticle

Development and Validation of an HPLC–PDA Method for Quality Control of Jwagwieum, an Herbal Medicine Prescription: Simultaneous Analysis of Nine Marker Compounds

by Chang-Seob Seo, Jeeyoun Jung and Sarah Shin

Pharmaceuticals 2025, 18(4), 481; https://doi.org/10.3390/ph18040481 (registering DOI) - 27 Mar 2025

Abstract

Background/Objectives: Jwagwieum (or Joa-Gui Em; JGE) consists of six herbal medicines, Rehmannia glutinosa (Gaertn.) DC., Dioscorea japonica Thunb., Lycium chinense Mill., Cornus officinalis Siebold & Zucc., Poria cocos Wolf, and Glycyrrhiza uralensis Fisch., and has been widely used to treat kidney-yin deficiency [...] Read more.

Background/Objectives: Jwagwieum (or Joa-Gui Em; JGE) consists of six herbal medicines, Rehmannia glutinosa (Gaertn.) DC., Dioscorea japonica Thunb., Lycium chinense Mill., Cornus officinalis Siebold & Zucc., Poria cocos Wolf, and Glycyrrhiza uralensis Fisch., and has been widely used to treat kidney-yin deficiency syndrome. In the present study, a high-performance liquid chromatography with photodiode array detector (HPLC–PDA) method for the simultaneous quantification of the nine components, i.e., gallic acid, 5-(hydroxymethyl)furfural, morroniside, loganin, liquiritin apioside, liquiritin, ononin, glycyrrhizin, and allantoin, was developed. Methods: The developed HPLC–PDA assay for quality control of JGE was validated with respect to linearity, limit of detection (LOD), limit of quantification (LOQ), recovery, and precision. Results: In the regression equation of the calibration curve, the coefficient of determination was ≥ 0.9980, and LOD and LOQ were 0.003–0.071 μg/mL and 0.010–0.216 μg/mL, respectively. Recovery and precision (relative standard deviation) were 96.36–106.95% and < 1.20%, respectively. In this analytical method, nine compounds were detected at concentrations of 0.15–3.69 mg/lyophilized gram. Conclusions: The developed and validated analytical method could be used to obtain basic data for the quality control of JGE and related herbal prescriptions. Full article

(This article belongs to the Special Issue Natural Pharmaceutical Component Analysis)

14 pages, 539 KiB

Open AccessArticle

The Combined Treatment of Chinese Herbal Medicines Is Correlated with a Lower Risk of Rheumatoid Arthritis in Patients with Depression: Evidence from a Population-Based Patient–Control Study

by Chieh-Tsung Yen, Hanoch Livneh, Hui-Ju Huang, Ming-Chi Lu, Wei-Jen Chen and Tzung-Yi Tsai

Pharmaceuticals 2025, 18(4), 480; https://doi.org/10.3390/ph18040480 (registering DOI) - 27 Mar 2025

Abstract

Background: Major depression places psychological strain on the individual that may increase the risk of developing rheumatoid arthritis (RA). Though the use of Chinese herbal medicines (CHMs) is widespread in clinical practice, its effect on the prevention of RA incidents is still unknown. [...] Read more.

Background: Major depression places psychological strain on the individual that may increase the risk of developing rheumatoid arthritis (RA). Though the use of Chinese herbal medicines (CHMs) is widespread in clinical practice, its effect on the prevention of RA incidents is still unknown. This study aimed to evaluate the association between CHMs use by patients with depression and their subsequent risk of being diagnosed with RA. Methods: This nested case–control study used claims data from a nationwide insurance database. We identified patients aged 20–70 years with newly diagnosed depression and without pre-existing RA between 2002 and 2010. We enrolled those with RA onset occurring after depression by the end of 2013 (n = 973). Randomly matched controls were selected from the remaining patients with depression but without RA (n = 1946). Conditional logistic regression analysis was executed to assess the association between CHMs use and RA onset. Data are presented as p-values with the significance set at 0.05 and as odds ratios (ORs) with 95% confidence intervals (CIs). Results: In this study, we found that adding CHMs treatment to conventional antidepressants greatly decreased the subsequent risk of RA among patients with depression, with an ORs of 0.64 (95% CIs: 0.57–0.76). Those using CHMs for more than three years had the most striking benefit, with a 61% lower risk of RA. Notably, initiating CHMs within the first 2 years after depression onset resulted in the greatest decrease in the RA risk. Conclusion: Using CHMs with conventional antidepressant therapy reduced the RA risk among patients with depression. Further well-designed randomized controlled trials are needed to determine the molecular mechanism underlying the action of these herbal agents. Full article

(This article belongs to the Special Issue Natural Products as an Alternative for Treatment of Human Diseases)

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23 pages, 12122 KiB

Open AccessArticle

Innovative Application of Medicinal Insects: Employing UHPLC-MS, Bioinformatics, In Silico Studies and In Vitro Experiments to Elucidate the Multi-Target Hemostatic Mechanism of Glenea cantor (Coleoptera: Cerambycidae) Charcoal-Based Medicine

by Bangyu Zhong, Wen Zhang, Liangshan Ming, Qimeng Fan, Lei Zhang, Hongyu Lai, Genwang Huang, Hongning Liu and Zishu Dong

Pharmaceuticals 2025, 18(4), 479; https://doi.org/10.3390/ph18040479 (registering DOI) - 27 Mar 2025

Abstract

Background: Longhorn beetles, a widely recognized group of Chinese traditional medicinal insects, are characterized by their notable hemostatic properties. However, the comprehensive understanding of their medicinal potential has been hindered by the limitations of current research methodologies. Methods: This study focuses on the [...] Read more.

Background: Longhorn beetles, a widely recognized group of Chinese traditional medicinal insects, are characterized by their notable hemostatic properties. However, the comprehensive understanding of their medicinal potential has been hindered by the limitations of current research methodologies. Methods: This study focuses on the species Glenea cantor (Fabricius), which can produce several generations per year, and introduces a novel method using microwave carbonization techniques. By employing an in vitro coagulation test, UHPLC-MS, network pharmacology, molecular docking, and molecular dynamics simulation, the hemostatic efficacy and mechanism of action of Glenea cantor charcoal medicine (GC-CM) were thoroughly studied. Results: In vitro coagulation tests showed that GC-CM significantly reduced the activated partial thromboplastin time (APTT) and prothrombin time (PT), indicating its ability to enhance the coagulation cascade and preliminarily confirming its hemostatic efficacy (p < 0.01 vs. blank control group). The analysis revealed that GC-CM comprises 453 components, including 137 bioactive components with high human utilization. After predictions via databases such as SwissTargetPrediction and deduplication, 215 targets linked to hemostatic specificity were identified. These targets regulate signaling pathways such as platelet activation, complement and coagulation cascades, and cGMP-PKG. Molecular docking demonstrated strong affinities between key targets such as SRC and PIK3R1 and compounds such as 2′,6′-dihydroxy 4′-methoxydihydrochalcone, and 1-monolinoleoyl-rac-glycerol (binding energy < −5 kcal/mol). Molecular dynamics simulations show good binding capacity between core components and targets Conclusions: The aim of this study was to elucidate the material basis and mechanism of the hemostatic efficacy of GC-CM, offering a model for exploring other insect-based medicinal resources. Full article

(This article belongs to the Section Natural Products)

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24 pages, 4583 KiB

Open AccessReview

Targeting the Endocannabinoidome: A Novel Approach to Managing Extraintestinal Complications in Inflammatory Bowel Disease

by Dinesh Thapa, Anjali Ghimire, Leon N. Warne and Rodrigo Carlessi

Pharmaceuticals 2025, 18(4), 478; https://doi.org/10.3390/ph18040478 (registering DOI) - 27 Mar 2025

Abstract

Background: Inflammatory bowel disease (IBD) is a chronic inflammatory disorder marked by persistent gastrointestinal inflammation and a spectrum of systemic effects, including extraintestinal manifestations (EIMs) that impact the joints, skin, liver, and eyes. Conventional therapies primarily target intestinal inflammation, yet they frequently fail [...] Read more.

Background: Inflammatory bowel disease (IBD) is a chronic inflammatory disorder marked by persistent gastrointestinal inflammation and a spectrum of systemic effects, including extraintestinal manifestations (EIMs) that impact the joints, skin, liver, and eyes. Conventional therapies primarily target intestinal inflammation, yet they frequently fail to ameliorate these systemic complications. Recent investigations have highlighted the complex interplay among the immune system, gut, and nervous system in IBD pathogenesis, thereby underscoring the need for innovative therapeutic approaches. Methods: We conducted a comprehensive literature search using databases such as PubMed, Scopus, Web of Science, Science Direct, and Google Scholar. Keywords including “cannabinoids”, “endocannabinoid system”, “endocannabinoidome”, “inflammatory bowel disease”, and “extraintestinal manifestations” were used to identify peer-reviewed original research and review articles that explore the role of the endocannabinoidome (eCBome) in IBD. Results: Emerging evidence suggests that eCBome—a network comprising lipid mediators, receptors (e.g., CB1, CB2, GPR55, GPR35, PPARα, TRPV1), and metabolic enzymes—plays a critical role in modulating immune responses, maintaining gut barrier integrity, and regulating systemic inflammation. Targeting eCBome not only improves intestinal inflammation but also appears to mitigate metabolic, neurological, and extraintestinal complications such as arthritis, liver dysfunction, and dermatological disorders. Conclusions: Modulation of eCBome represents a promising strategy for comprehensive IBD management by addressing both local and systemic disease components. These findings advocate for further mechanistic studies to develop targeted interventions that leverage eCBome as a novel therapeutic avenue in IBD. Full article

(This article belongs to the Special Issue Therapeutic Potential of Cannabinoid Receptors Type 1 and 2—Novel Insights for Enhancing the Chance of Clinical Success)

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14 pages, 975 KiB

Open AccessArticle

Using Physiologically Based Pharmacokinetic Models for Assessing Pharmacokinetic Drug–Drug Interactions in Patients with Chronic Heart Failure Taking Narrow Therapeutic Window Drugs

by Nadezhda Hvarchanova, Maya Radeva-Ilieva and Kaloyan D. Georgiev

Pharmaceuticals 2025, 18(4), 477; https://doi.org/10.3390/ph18040477 (registering DOI) - 27 Mar 2025

Abstract

Background: Pharmacotherapy of chronic heart failure (CHF) with a reduced ejection fraction includes a combination of drugs. Often, different groups of drugs are added together for the treatment of concomitant conditions, such as statins, anticoagulants, antiplatelet agents, and cardiac glycosides, which have a [...] Read more.

Background: Pharmacotherapy of chronic heart failure (CHF) with a reduced ejection fraction includes a combination of drugs. Often, different groups of drugs are added together for the treatment of concomitant conditions, such as statins, anticoagulants, antiplatelet agents, and cardiac glycosides, which have a narrow therapeutic window. Increased medication intake is a prerequisite for the increased risk of potential adverse drug–drug interactions (DDI), especially those occurring at the pharmacokinetic level. The main objectives of this study are to identify the most common potential pharmacokinetic drug–drug interactions (pPKDDIs), to explore more complex cases, and to simulate and analyze them with appropriate software. Methods: The data selected for the simulations were collected over a two-year period from January 2014 to December 2015. Identification of the pPKDDIs was carried out using Lexicomp Drug interaction, while simulations were performed with Simcyp software (V20, R1). Results: The most common pharmacokinetic mechanisms responsible for the occurrence of drug–drug interactions in the selected drugs with narrow therapeutic windows are those featuring the cytochrome isoforms CYP3A4 and 2C9 and the efflux pump—P-glycoprotein (P-gp). Simulations with the available data in Simcyp software showed possibilities to analyze and evaluate pPKDDIs, which would be difficult to assess without appropriate software, as well as ways to manage them. Conclusions: The frequency and complexity of pPKDDIs in patients with cardiovascular disease are high. Therefore, such patients require a specific approach to reduce these risks as well as to optimize the therapy. An appropriate PBPK software with the necessary database would be suitable in these cases. Full article

(This article belongs to the Special Issue Pharmacovigilance in Drug Therapy: Drug–Drug Interactions and Safety Evaluation, 2nd Edition)

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17 pages, 3728 KiB

Open AccessArticle

Further In Vitro and Ex Vivo Pharmacological and Kinetic Characterizations of CCF219B: A Positive Allosteric Modulator of the α_1A-Adrenergic Receptor

by Robert S. Papay and Dianne M. Perez

Pharmaceuticals 2025, 18(4), 476; https://doi.org/10.3390/ph18040476 (registering DOI) - 27 Mar 2025

Abstract

Background: Alterations in the adrenergic system have been associated with the pathophysiology of Alzheimer’s disease (AD). A novel α_1A-adrenergic receptor (AR)-positive allosteric modulator (PAM), CCF219B, has been shown to outperform donepezil with rescue of AD cognition/memory deficits with a reduction in [...] Read more.

Background: Alterations in the adrenergic system have been associated with the pathophysiology of Alzheimer’s disease (AD). A novel α_1A-adrenergic receptor (AR)-positive allosteric modulator (PAM), CCF219B, has been shown to outperform donepezil with rescue of AD cognition/memory deficits with a reduction in amyloid biomarkers and without cardiovascular side effects. Initial pharmacological analysis in transfected cell lines revealed a signal bias with increased efficacy (but not potency) of cAMP signaling and ligand selectivity for norepinephrine (NE). As most GPCR allosteric modulators change the potency of agonists, we hypothesized and now report that CCF219B induced additional aspects of its allosteric interactions with NE that may provide mechanistic insight. Methods: Using Rat-1 fibroblasts stably transfected with α_1A-AR, we determined the activation profile of pERK and p38 messengers by CCF219B in the presence of NE. Using membranes prepared from the stably transfected fibroblasts or from the brain of WT mice or the AD mouse model, hAPP(lon), equilibrium or kinetic radioligand-binding analyses were performed. Results: We identified p-ERK1/2 but not p38 as an additional signal pathway that is potentiated by CCF219B in the presence of NE. An analysis of binding studies of CCF219B in membranes derived from the brains of WT or hAPP(lon) mice revealed profiles that were time-dependent and resulted in an increase in α_1A-AR expression that was unaltered in the presence of cycloheximide or when performed at 37 °C. hAPP(lon) mice displayed a reduction in α_1A-AR-binding sites that were rescued upon prolonged incubation with CCF219B but also displayed a compensatory increase in α_1B/D-AR subtype expression. Binding kinetics reveal that CCF219B can decrease the association rate of ³H-NE but only in the presence of GTP. The association rate increased for the radiolabeled antagonist, ¹²⁵I-HEAT. There were no changes in the dissociation rate of either radiolabel. Conclusions: CCF219B affects the association but not the dissociation rate of NE and explains its ability to increase the active state of the receptor by promoting a pre-coupled conformation, consistent with increasing efficacy but not potency. Potentiation of pERK may contribute to CCF219B’s ability to confer neuroprotection and be pro-cognitive in AD. CCF219B’s ability to increase the expression of α_1A-AR provides a positive feedback loop and strengthens the hypothesis that α₁-AR subtypes may be involved in AD etiology and/or progression. Full article

(This article belongs to the Special Issue Pharmacotherapy for Alzheimer’s Disease)

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30 pages, 5575 KiB

Open AccessArticle

Regulation of Intestinal Barrier Function and Gut Microbiota by Hot Melt Extrusion-Drug Delivery System-Prepared Mulberry Anthocyanin in an Inflammatory Bowel Disease Model

by Eun-Ji Go, Byeong Ryeol Ryu, Gyeong Ju Gim, Ye Rim Shin, Min Ji Kang, Min Jun Kim, Jong-Suep Baek and Jung Dae Lim

Pharmaceuticals 2025, 18(4), 475; https://doi.org/10.3390/ph18040475 (registering DOI) - 27 Mar 2025

Abstract

Background/Objectives: Anthocyanins (ACNs) derived from mulberry (Morus alba L.) exhibit potent antioxidant and anti-inflammatory activities. However, their low stability and bioavailability in physiological environments limit their therapeutic potential. This study aimed to enhance the stability and controlled release ACNs using a hot-melt [...] Read more.

Background/Objectives: Anthocyanins (ACNs) derived from mulberry (Morus alba L.) exhibit potent antioxidant and anti-inflammatory activities. However, their low stability and bioavailability in physiological environments limit their therapeutic potential. This study aimed to enhance the stability and controlled release ACNs using a hot-melt extrusion drug delivery system (HME-DDS) formulation, HME-MUL-F2, and evaluate its effects on gut barrier function and microbiota composition in a DSS-induced colitis model. Methods: The anthocyanin content of HME-MUL-F2 was quantified and compared with that of raw mulberry extract. The formulation’s protective effects were assessed in Caco-2 and RAW 264.7 cells, confirming its biocompatibility and anti-inflammatory properties. The therapeutic efficacy was further evaluated in a dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) model, focusing on gut barrier integrity, inflammatory cytokine modulation, and gut microbiota composition. Results: HME-MUL-F2 significantly improved gut barrier function by upregulating tight junction proteins and reducing inflammatory cytokine levels in the colitis model. Moreover, the formulation modulated gut microbiota composition, promoting beneficial bacteria while suppressing pathogenic strains. HME-MUL-F2 administration led to a significant increase in the Bacteroidetes-to-Firmicutes ratio, which is associated with improved gut health. These results indicate that HME-MUL-F2 significantly enhances anthocyanin bioavailability, leading to improved gut health and potential therapeutic applications for inflammatory conditions. Conclusions: This study highlights the potential of HME technology for improving the stability, bioavailability, and therapeutic efficacy of anthocyanins. HME-MUL-F2 is a sustained-release formulation that enhances gut barrier function and modulates intestinal microbial balance in a DSS-induced inflammatory bowel disease model. These findings strongly suggest that the observed therapeutic effects of HME-MUL-F2 are primarily due to enhanced anthocyanin bioavailability and targeted delivery to the colon, although further clinical studies will provide more definitive confirmation. Full article

(This article belongs to the Special Issue Pharmacological Activities of Flavonoids and Their Analogues 2024)

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17 pages, 6778 KiB

Open AccessArticle

Comprehensive Studies on the Regulation of Type 2 Diabetes by Cucurbitane-Type Triterpenoids in Momordica charantia L.: Insights from Network Pharmacology and Molecular Docking and Dynamics

by Yang Niu, Peihang Li and Zongran Pang

Pharmaceuticals 2025, 18(4), 474; https://doi.org/10.3390/ph18040474 (registering DOI) - 27 Mar 2025

Abstract

Background/Objectives: Momordica charantia L. (M. charantia), a widely cultivated and frequently consumed medicinal plant, is utilized in traditional medicine. Cucurbitane-type triterpenoids, significant saponin components of M. charantia, exhibit hypoglycemic effects; however, the underlying mechanisms remain unclear. Methods: This study [...] Read more.

Background/Objectives: Momordica charantia L. (M. charantia), a widely cultivated and frequently consumed medicinal plant, is utilized in traditional medicine. Cucurbitane-type triterpenoids, significant saponin components of M. charantia, exhibit hypoglycemic effects; however, the underlying mechanisms remain unclear. Methods: This study utilized comprehensive network pharmacology to identify potential components of M. charantia cucurbitane-type triterpenoids that may influence type 2 diabetes mellitus (T2DM). Additionally, molecular docking and molecular dynamics studies were performed to assess the stability of the interactions between the selected components and key targets. Results: In total, 22 candidate active components of M. charantia cucurbitane-type triterpenoids and 1165 disease targets for T2DM were identified through database screening. Molecular docking and molecular dynamics simulations were conducted for five key components (Kuguacin J, 25-O-methylkaravilagenin D, Momordicine I, momordic acid, and Kuguacin S) and three key targets (AKT1, IL6, and SRC), and the results demonstrated stable binding. The experimental results indicate that the interactions between momordic acid-AKT1 and momordic acid-IL6 are stable. Conclusions: Momordic acid may play a crucial role in M. charantia’s regulation of T2DM, and AKT1 and IL6 seem to be key targets for the therapeutic action of M. charantia in managing T2DM. Full article

(This article belongs to the Section Pharmacology)

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15 pages, 4294 KiB

Open AccessArticle

Synthesis and Reactivity of Oligo(ethylene glycol)-Tethered Morita–Baylis–Hillman Dimers in the Formation of Macrocyclic Structures Showing Remarkable Cytotoxicity

by Marco Paolino, Mario Saletti, Jacopo Venditti, Arianna Zacchei, Alessandro Donati, Claudia Bonechi, Germano Giuliani, Stefania Lamponi and Andrea Cappelli

Pharmaceuticals 2025, 18(4), 473; https://doi.org/10.3390/ph18040473 (registering DOI) - 27 Mar 2025

Abstract

Background/Objectives: Crown ethers have received increasing interest owing to their ability to form stable complexes with cations. This molecular feature has been successfully exploited in the development of biologically relevant ionophores. Methods: In order to obtain innovative crown ethers derivatives, a [...] Read more.

Background/Objectives: Crown ethers have received increasing interest owing to their ability to form stable complexes with cations. This molecular feature has been successfully exploited in the development of biologically relevant ionophores. Methods: In order to obtain innovative crown ethers derivatives, a Morita–Baylis–Hillman adduct (MBHA) acetate (4) bearing a phenylacetylene moiety was dimerized via the click-chemistry CuAAC reaction with oligo(ethylene glycol) diazide derivatives to build-up a small series of dimeric MBHA derivatives (5a-d). These dimeric MBHA derivatives were reacted with n-butylamine to afford tunable macrocyclic crown ether-paracyclophane hybrid architectures (6a-d). Results: Compounds (E,Z)-6a, (E,E)-6a, 6b-d showed, in human breast cancer MDA-MB-231 and human melanoma A375 cells, IC₅₀ values comparable with those of reference anticancer agent Doxorubicin. Conclusions: This exploration approach provides original new macrocyclic architectures potentially useful as anticancer agents. Full article

(This article belongs to the Section Medicinal Chemistry)

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15 pages, 4975 KiB

Open AccessArticle

Paeonol Suppresses Bladder Cancer Progression via Apoptotic Pathways: Insights from In Vitro and In Vivo Studies

by Lu Ying, Ruolan Chen, Rui Guo, Youfeng Liang, Mingxuan Hao, Xiaoyang Chen, Wenjing Zhang, Changyuan Yu and Zhao Yang

Pharmaceuticals 2025, 18(4), 472; https://doi.org/10.3390/ph18040472 (registering DOI) - 27 Mar 2025

Abstract

Background/Objectives: Bladder cancer (BC), a highly heterogeneous and mutation-prone malignancy, remains a significant therapeutic challenge due to its propensity for recurrence, metastasis, and drug resistance. Natural products, particularly paeonol, a bioactive compound derived from Moutan Cortex in traditional Chinese medicine, have shown [...] Read more.

Background/Objectives: Bladder cancer (BC), a highly heterogeneous and mutation-prone malignancy, remains a significant therapeutic challenge due to its propensity for recurrence, metastasis, and drug resistance. Natural products, particularly paeonol, a bioactive compound derived from Moutan Cortex in traditional Chinese medicine, have shown promising potential in cancer therapy. This study aims to evaluate the anti-BC effects of paeonol and elucidate its underlying molecular mechanisms. Methods: In vitro experiments were conducted using T24 and J82 BC cell lines to assess paeonol’s effects on cell viability, migration, apoptosis, and cell cycle progression via CCK-8, scratch, flow cytometry, RT-qPCR, and Western blot analyses. In vivo efficacy was evaluated using a xenograft mouse model, with tumor growth monitored and histopathological analysis performed. Results: Paeonol significantly inhibited BC cell proliferation and migration in a dose- and time-dependent manner, with IC₅₀ values of 225 μg/mL (T24) and 124 μg/mL (J82) at 48 h. It induced apoptosis and arrested the cell cycle at the G1 phase, accompanied by upregulation of pro-apoptotic proteins (BID, BAX, BIM, and p53). In vivo, paeonol reduced tumor volume and weight without histopathological abnormalities in vital organs. Conclusions: Paeonol exhibits potent anti-BC activity by apoptotic pathways and by arresting the cell cycle at the G1 phase and inhibiting tumor growth. Its favorable safety profile and multi-target mechanisms highlight its potential as a promising therapeutic candidate for BC. These findings provide a foundation for further clinical development of paeonol-based therapies. Full article

(This article belongs to the Section Pharmacology)

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18 pages, 4555 KiB

Open AccessArticle

AHR Agonist ITE Boosted PD1 Antibody’s Effects by Inhibiting Myeloid-Derived Cells Suppressive Cells in an Orthotopic Mouse Glioma Model

by Pei Gong, Lijiao Zhao, Yunlong Ma, Qiuting Shu, Hui Sun, Jing Lu, Fanhua Meng and Fang Wan

Pharmaceuticals 2025, 18(4), 471; https://doi.org/10.3390/ph18040471 (registering DOI) - 27 Mar 2025

Abstract

Background: Glioblastoma is “cold”. Consequently, immune checkpoint blockade therapy has failed to improve patients‘ survival, which is negatively correlated with patients’ peripheral MDSC counts. AHR is known to mediate immune-suppressive functions of certain tryptophan metabolites such as kynurenine; yet, there lack of reports [...] Read more.

Background: Glioblastoma is “cold”. Consequently, immune checkpoint blockade therapy has failed to improve patients‘ survival, which is negatively correlated with patients’ peripheral MDSC counts. AHR is known to mediate immune-suppressive functions of certain tryptophan metabolites such as kynurenine; yet, there lack of reports on how AHR agonists affect glioma immunity. Methods/Objectives: We hypothesized that ITE could synergize with PD1 antibody as AHR is one major node of immune-suppressive pathways, and tested it using an immune-competent mouse glioma model. Results: The combination of ITE+PD1 antibody glioma MDSC was significantly reduced, along with increased infiltration of the CD4−CD8+ and CD4+CD8+ T cells, leading to extended mouse survival. ITE treatment alone significantly reduces the infiltration of CD11b+Ly6G+Ly6Clo cells, namely PMN-MDSCs, and neutrophils, while the combination with PD1 antibody significantly reduces all MDSCs plus neutrophils. The presence of ITE inhibits the expression of IL11 and the in vitro induction of MDSCs from mouse PBMCs by IL11. The identification of the ITE-AHR-IL11-MDSC pathway provides more mechanistic insights into AHR’s effects. The fact that ITE, which is otherwise immune-suppressive, can activate immunity in glioma indicates that searching for drugs targeting AHR should go beyond antagonists. Full article

(This article belongs to the Section Medicinal Chemistry)

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20 pages, 5127 KiB

Open AccessArticle

An Experimental Design Approach for Producing Curcumin-Loaded Solid Lipid Nanoparticles

by Ongun Mehmet Saka, Cemre İrem Aygüler, Neval Sevinç Özdemir, Bilge Sürücü, Egemen Çakırlı, Emirhan Nemutlu and Gülen Melike Demirbolat

Pharmaceuticals 2025, 18(4), 470; https://doi.org/10.3390/ph18040470 (registering DOI) - 27 Mar 2025

Abstract

Background/Objectives: Curcumin has well-established efficacy in a variety of disorders due to its prominent antioxidant, antiaging, anti-inflammatory, chemosensitizing, and anticancer activities. Despite its numerous benefits, curcumin exhibits low bioavailability mainly due to its poor solubility, poor absorption, rapid metabolism, and quick excretion, [...] Read more.

Background/Objectives: Curcumin has well-established efficacy in a variety of disorders due to its prominent antioxidant, antiaging, anti-inflammatory, chemosensitizing, and anticancer activities. Despite its numerous benefits, curcumin exhibits low bioavailability mainly due to its poor solubility, poor absorption, rapid metabolism, and quick excretion, consequently limiting its clinical applications. In this study, we investigated the most convenient ingredients in SLNs to enhance curcumin’s solubility by examining the effects of multiple independent variables simultaneously using an experimental design. Methods: After curcumin’s saturation solubility was investigated, SLN formulations were produced. The optimum formulation was determined with the help of experimental design. The SLNs were characterized in terms of the particle size and distribution, zeta potential, shape, entrapment efficiency, drug loading capacity, and drug release. The cell viability and cell internalization were also evaluated. Results: An impressive synergistic effect was achieved with the combination of Brij and Gelucire 48/16, which increased curcumin’s solubility in water by 452.5 times. Curcumin-loaded SLNs were successfully produced with a spherical shape and particle size of 389.3 ± 9.95 nm. The encapsulation efficiency was directly proportionate to the amount of curcumin and the stirring speed. Curcumin in the SLNs entered the cancer cells more easily than curcumin alone. Conclusions: Our results demonstrate that the quantity of surfactant is a significant factor influencing the efficiency of drug loading. Finally, the 3:1 (Brij–Gelucire48/16) ratio markedly enhanced the loading efficiency. The cellular internalization and, consequently, the anticancer efficacy against adenocarcinomic human alveolar basal epithelial cells were improved with SLNs. This could be a promising approach for lipid-based colloidal drug delivery systems. Full article

(This article belongs to the Section Pharmaceutical Technology)

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18 pages, 2090 KiB

Open AccessArticle

Is Copper‑61 the New Gallium‑68? Automation and Preclinical Proof‑of‑Concept of ⁶¹Cu‑Based Radiopharmaceuticals for Prostate Cancer Imaging

by Diana Rodrigues, Alexandra I. Fonseca, Sérgio do Carmo, José Sereno, Ivanna Hrynchak, João N. Moreira, Célia Gomes and Antero Abrunhosa

Pharmaceuticals 2025, 18(4), 469; https://doi.org/10.3390/ph18040469 (registering DOI) - 26 Mar 2025

Abstract

Background: While gallium‑68 has traditionally dominated PET imaging in oncology, copper radionuclides have sparked interest for their potential applications in nuclear medicine and theranostics. Considering the advantageous physical decay properties of copper‑61 compared to those of gallium‑68, we describe a fully automated GMP‑compliant [...] Read more.

Background: While gallium‑68 has traditionally dominated PET imaging in oncology, copper radionuclides have sparked interest for their potential applications in nuclear medicine and theranostics. Considering the advantageous physical decay properties of copper‑61 compared to those of gallium‑68, we describe a fully automated GMP‑compliant synthesis process for ⁶¹Cu‑based radiopharmaceuticals and demonstrate their in vivo application for targeting the overexpressed PSMA by PET/MR imaging. Methods: Copper‑61 was obtained through the irradiation of natural zinc liquid targets in a biomedical cyclotron. [⁶¹Cu]Cu‑DOTAGA‑PSMA‑I&T and [⁶¹Cu]Cu‑NODAGA‑PSMA‑I&T were produced without manual intervention in two Synthera^® Extension modules. Radiochemical purity was analyzed by radio‑HPLC and iTLC. Cellular uptake was evaluated in LNCaP and DU145 cells. In vivo PET/MRI was performed in control mice to evaluate the biodistribution of both radiopharmaceuticals, and in tumor‑bearing mice to assess the targeting ability towards PSMA. Results: The fully automated process developed proved to be effective for the synthesis of ⁶¹Cu‑based radiopharmaceuticals, with appropriate molar activities. The final products exhibited high radiochemical purity (>98%) and remained stable for up to 6 h after the EOS. A time‑dependent increase in cellular uptake was observed in LNCaP cells, but not in DU145 cells. As opposed to [⁶¹Cu]Cu‑NODAGA‑PSMA‑I&T, [⁶¹Cu]Cu‑DOTAGA‑PSMA‑I&T exhibited poor kinetic stability in vivo. Subsequent PET/MR imaging with [⁶¹Cu]Cu‑NODAGA‑PSMA‑I&T showed tumor uptake lasting up to 4 h post‑injection, predominant renal clearance, and no detectable accumulation in non‑targeted organs. Conclusions: These results demonstrate the feasibility of the implemented process, which yields adequate amounts of high‑quality radiopharmaceuticals and can be adapted to any standard production facility. This streamlined approach enhances reproducibility and scalability, bringing copper‑61 closer to widespread clinical use, to the detriment of the conventionally accepted gallium‑68. Full article

(This article belongs to the Special Issue Past, Present and Future Radiotracer Techniques: Radiopharmaceuticals in Cancer Theranostics)

24 pages, 11576 KiB

Open AccessArticle

Aromatic Molecular Compatibility Attenuates Influenza Virus-Induced Acute Lung Injury via the Lung–Gut Axis and Lipid Droplet Modulation

by Yi Li, Jiakang Jiao, Haoyi Qiao, Conghui Wang, Linze Li, Fengyu Jin, Danni Ye, Yawen Chen, Qi Zhang, Min Li, Zhongpeng Zhao, Jianjun Zhang and Linyuan Wang

Pharmaceuticals 2025, 18(4), 468; https://doi.org/10.3390/ph18040468 (registering DOI) - 26 Mar 2025

Abstract

Background: Acute lung injury (ALI) is a major cause of death in patients with various viral pneumonias. Our team previously identified four volatile compounds from aromatic Chinese medicines. Based on molecular compatibility theory, we defined their combination as aromatic molecular compatibility (AC), though [...] Read more.

Background: Acute lung injury (ALI) is a major cause of death in patients with various viral pneumonias. Our team previously identified four volatile compounds from aromatic Chinese medicines. Based on molecular compatibility theory, we defined their combination as aromatic molecular compatibility (AC), though its therapeutic effects and underlying mechanisms remain unclear. Methods: This study used influenza A virus (IAV) A/PR/8/34 to construct cell and mouse models of ALI to explore AC’s protective effects against viral infection. The therapeutic effect of AC was verified by evaluating the antiviral efficacy in the mouse models, including improvements in their lung and colon inflammation, oxidative stress, and the suppression of the NLRP3 inflammasome. In addition, 16S rDNA and lipid metabolomics were used to analyze the potential therapeutic mechanisms of AC. Results: Our in vitro and in vivo studies demonstrated that AC increased the survival of the IAV-infected cells and mice, inhibited influenza virus replication and the expression of proinflammatory factors in the lung tissues, and ameliorated barrier damage in the colonic tissues. In addition, AC inhibited the expression of ROS and the NLRP3 inflammasome and improved the inflammatory cell infiltration into the lung tissues. Finally, AC effectively regulated intestinal flora disorders and lipid metabolism in the model mice, significantly reduced cholesterol and triglyceride expression, and thus reduced the abnormal accumulation of lipid droplets (LDs) after IAV infection. Conclusions: In this study, we demonstrated that AC could treat IAV-induced ALIs through multiple pathways, including antiviral and anti-inflammatory pathways and modulation of the intestinal flora and the accumulation of LDs. Full article

(This article belongs to the Section Natural Products)

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20 pages, 363 KiB

Open AccessReview

The Mechanisms of Lithium Action: The Old and New Findings

by Kosma Sakrajda and Janusz K. Rybakowski

Pharmaceuticals 2025, 18(4), 467; https://doi.org/10.3390/ph18040467 - 26 Mar 2025

Abstract

Despite lithium’s presence in modern psychiatry for three-quarters of a century, the mechanisms of its therapeutic action have not been fully elucidated. This article presents the evolution of the views on these mechanisms, and both the old and new findings are discussed. Among [...] Read more.

Despite lithium’s presence in modern psychiatry for three-quarters of a century, the mechanisms of its therapeutic action have not been fully elucidated. This article presents the evolution of the views on these mechanisms, and both the old and new findings are discussed. Among the old mechanisms, lithium’s effect on the purinergic system; electrolyte metabolism; membrane transport; and second messenger systems, namely, cyclic nucleotide and phosphatidylinositol (PI), glycogen synthase kinase-3beta (GSK-3β), brain-derived neurotrophic factor, and neurotransmitters, are discussed. The new data were obtained from in vitro studies, molecular biology, and genetic research. They showed the effects of lithium on the immune system, biological rhythms, telomere functions, and mitochondria. In this article, each lithium mechanism is considered in the light of its association with the pathogenesis of bipolar disorder or/and as a marker of the lithium response. Although not exhaustive, this review elucidates the multiple potential mechanisms of lithium action. It was also observed that many seemingly “old” mechanisms have experienced a resurgence in research conducted during the 21st century. Additionally, many studies converged on the previously postulated mechanisms of lithium inhibiting GSK-3β and PI. Full article

(This article belongs to the Special Issue Lithium in Psychiatric Therapy: Celebrating 75th Anniversary)

17 pages, 3398 KiB

Open AccessArticle

Enhancing Boron Neutron Capture Therapy (BNCT) with Materials Based on COSAN-Functionalized Nanoparticles

by Albert Ferrer-Ugalde, Amanda Muñoz-Juan, Anna Laromaine, Paula Curotto, Susana Nievas, María Alejandra Dagrosa, Marcos Couto and Rosario Núñez

Pharmaceuticals 2025, 18(4), 466; https://doi.org/10.3390/ph18040466 - 26 Mar 2025

Abstract

Background/Objectives: Boron neutron capture therapy (BNCT) is a promising approach for selectively targeting and destroying malignant cells using ¹⁰B isotopes. A significant challenge in BNCT lies in the development of efficient boron delivery systems that ensure adequate boron accumulation within tumor [...] Read more.

Background/Objectives: Boron neutron capture therapy (BNCT) is a promising approach for selectively targeting and destroying malignant cells using ¹⁰B isotopes. A significant challenge in BNCT lies in the development of efficient boron delivery systems that ensure adequate boron accumulation within tumor cells. This study aims to synthesize, characterize, and evaluate COSAN-functionalized nanoparticles (NP@I-COSAN) as a potential boron carrier for BNCT. Methods: Hybrid nanoparticles were synthesized by conjugating monoiodinated cobaltabisdicarbollides (I-COSAN) to commercially available acrylic polymer-based nanoparticles. Functionalization and cellular uptake were confirmed through FTIR, TGA, UV-Vis spectroscopy, and TEM/EDX analyses. Biocompatibility was evaluated by assessing cytotoxicity in HeLa cells and C. elegans as an in vivo model. Intracellular boron uptake was quantified using ICP-MS, with results compared to those obtained with 4-borono-L-phenylalanine conjugated to fructose (f-BPA). An in vitro BNCT proof-of-concept assay was also performed to evaluate therapeutic efficacy. Results: NP@I-COSAN demonstrated low cytotoxicity and efficient internalization in cells. ICP-MS analysis revealed stable boron retention, comparable to traditional boron agents. The BNCT assay further showed that NP@I-COSAN was effective in inducing tumor cell apoptosis, even at lower boron concentrations than conventional treatments. Conclusions: These results underscore the potential of NP@I-COSAN as an effective boron delivery system for BNCT, offering a promising strategy to enhance boron accumulation within tumor cells and improve treatment efficacy. Full article

(This article belongs to the Special Issue Boron in Medicinal Chemistry: From Synthesis to Therapeutic Applications)

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13 pages, 3538 KiB

Open AccessArticle

A Novel LC-APCI-MS/MS Approach for the Trace Analysis of 3,4-Difluoronitrobenzene in Linezolid

by Yujin Lim, Aelim Kim, Eunyeong Shin and Hwangeui Cho

Pharmaceuticals 2025, 18(4), 465; https://doi.org/10.3390/ph18040465 - 26 Mar 2025

Abstract

Background/Objectives: Oxazolidinones are novel antimicrobial agents used to combat bacterial infections, particularly multidrug-resistant strains. However, the synthesis of oxazolidinone derivatives, such as linezolid, often involves the use of 3,4-difluoronitrobenzene (DFNB) as an initiator. Despite its effectiveness, residual DFNB in drug products raises [...] Read more.

Background/Objectives: Oxazolidinones are novel antimicrobial agents used to combat bacterial infections, particularly multidrug-resistant strains. However, the synthesis of oxazolidinone derivatives, such as linezolid, often involves the use of 3,4-difluoronitrobenzene (DFNB) as an initiator. Despite its effectiveness, residual DFNB in drug products raises significant health concerns due to its structural similarity to toxic and carcinogenic nitrobenzenes. This contamination is particularly concerning in pharmaceutical formulations, where it poses potential patient safety hazards. Therefore, strict concentration limits for this impurity are necessary. Methods: To ensure tight control of DFNB concentrations, this study established an 8.3 µg/g target limit. An advanced high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to overcome current limitations in detecting trace DFNB. Under negative atmospheric pressure chemical ionization (APCI) conditions, DFNB exhibited characteristic ion formations, including [M]^•− through electron capture and [M − F + O]⁻ via substitution reactions. The quantitative method utilizes MS/MS ion transitions of the substitution product while optimizing chromatographic and spectrometric parameters to enhance both sensitivity and specificity. Conclusions: Validation tests confirm the efficiency, precision, and accuracy of this method, with a low limit of quantification (LOQ) of 5 ng/mL (0.83 µg/g). This technique enables accurate detection and quantification of DFNB in linezolid active pharmaceutical ingredient (API) and various formulations, providing a reliable tool for quality control. This method ensures the safe use of linezolid by effectively monitoring and minimizing the risks associated with DFNB contamination. Full article

(This article belongs to the Section Pharmaceutical Technology)

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17 pages, 3317 KiB

Open AccessArticle

Exploration of Ultrasound-Enhanced Transdermal Delivery Efficiency and Anti-Inflammatory Effect of Rutin

by Qing Yue, Bingbing He, Zhenyu Guo, Ningtao Zhang, Mei Zhang and Yufeng Zhang

Pharmaceuticals 2025, 18(4), 464; https://doi.org/10.3390/ph18040464 - 26 Mar 2025

Abstract

Background: Rutin is a natural flavonoid extracted primarily from plants with anti-inflammatory and antioxidant properties, and it is highly valuable in the cosmetics industry. However, the poor transdermal permeability of rutin limits its application via transdermal administration. Previous studies have predominantly focused [...] Read more.

Background: Rutin is a natural flavonoid extracted primarily from plants with anti-inflammatory and antioxidant properties, and it is highly valuable in the cosmetics industry. However, the poor transdermal permeability of rutin limits its application via transdermal administration. Previous studies have predominantly focused on chemical methods for enhancing penetration. This study investigated the potential of ultrasound as a physical method by which to augment the transdermal absorption and anti-inflammatory effects of rutin. Method: Through in vitro diffusion experiments, we analyzed the effects of the ultrasonic frequency and intensity on percutaneous absorption. The optimal ultrasound parameters were determined based on the intradermal retention rate, which is defined as the proportion of intradermal retention to the total penetration. Parameters with higher retention rates were considered optimal. To validate the anti-inflammatory efficacy of rutin delivered using the ultrasound-assisted method, we employed a tape-stripping technique to induce inflammation in BALB/c nude mice. Eight mice were assigned to each treatment group: (A) self-repair (control group), (B) regular rutin treatment, and (C) ultrasound-assisted treatment. Results: The research findings indicate that ultrasound frequency and intensity of 1 MHz and 0.2 W/cm², as well as 3 MHz and 0.2 W/cm², result in the maximum proportion of rutin intradermal retention, exhibiting values 1.8 times (using porcine skin) and 2.63 times (using nude mouse skin) higher than those achieved without ultrasound, respectively. Group C showed the shortest recovery time and displayed complete skin barrier function restoration by the fourth day

(p < 0.05)

, whereas group A exhibited the slowest recovery. Conclusions: This study offers an innovative approach for the transdermal delivery of rutin to facilitate skin barrier function repair. Full article

(This article belongs to the Section Pharmaceutical Technology)

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20 pages, 2061 KiB

Open AccessArticle

5,7-Dihydroxy-4-Methylcoumarin as a Functional Compound for Skin Pigmentation and Human Skin Safety

by Ye-Jin Lee, Yang Xu and Chang-Gu Hyun

Pharmaceuticals 2025, 18(4), 463; https://doi.org/10.3390/ph18040463 - 25 Mar 2025

Abstract

Background/Objectives: This study aims to investigate the effects of 5,7-dihydroxy-4-methylcoumarin (5,7D-4MC) on melanogenesis in B16F10 murine melanoma cells and to evaluate its safety as a potential ingredient for functional cosmetics and therapeutic agents targeting pigmentation-related disorders. Method: The cytotoxicity of 5,7D-4MC was assessed [...] Read more.

Background/Objectives: This study aims to investigate the effects of 5,7-dihydroxy-4-methylcoumarin (5,7D-4MC) on melanogenesis in B16F10 murine melanoma cells and to evaluate its safety as a potential ingredient for functional cosmetics and therapeutic agents targeting pigmentation-related disorders. Method: The cytotoxicity of 5,7D-4MC was assessed using an MTT assay, and melanin content and tyrosinase activity were measured at different concentrations (25, 50, 100 µM). Western blot analyses were conducted to evaluate the expression of key melanogenesis-related proteins (TYR, TRP-1, TRP-2, and MITF) and to investigate the regulation of major signaling pathways, including PKA/cAMP, GSK3β, and PI3K/AKT. Additionally, a human primary skin irritation test was performed on 32 participants to assess the dermatological safety of 5,7D-4MC. Results: 5,7D-4MC did not affect cell viability at concentrations below 100 µM and significantly promoted melanin production in a dose-dependent manner. Tyrosinase activity and the expression levels of melanogenic proteins increased significantly following 5,7D-4MC treatment. PKA and GSK3β pathways were activated, while the PI3K/AKT pathway was downregulated. The skin irritation test showed that 5,7D-4MC exhibited low irritation potential at concentrations of 50 µM and 100 µM. Conclusions: 5,7D-4MC enhances melanogenesis and demonstrates low skin irritation, making it a promising candidate for therapeutic applications in treating hypopigmentation disorders, such as vitiligo, as well as a functional cosmetic ingredient. However, further studies involving human melanocytes and clinical trials are required to validate their efficacy. Full article

(This article belongs to the Special Issue Advancements in the Study of Biological Activities of Plant Extracts and Components)

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