Pharmaceuticals

13 pages, 1977 KiB

Open AccessArticle

Computational Screening and Experimental Evaluation of Wheat Proteases for Use in the Enzymatic Therapy of Gluten-Related Disorders

by Lyudmila V. Savvateeva, Olga E. Chepikova, Alena D. Solonkina, Artemiy A. Sakharov, Neonila V. Gorokhovets, Andrey V. Golovin and Andrey A. Zamyatnin, Jr.

Pharmaceuticals 2025, 18(4), 592; https://doi.org/10.3390/ph18040592 - 18 Apr 2025

Abstract

Background: Gluten-related disorders, particularly celiac disease, are triggered in susceptible individuals by the toxic effects of gluten, the major storage protein of wheat grains. This toxicity can be reduced by wheat glutenases. Members of the papain-like cysteine protease family, which can act in [...] Read more.

Background: Gluten-related disorders, particularly celiac disease, are triggered in susceptible individuals by the toxic effects of gluten, the major storage protein of wheat grains. This toxicity can be reduced by wheat glutenases. Members of the papain-like cysteine protease family, which can act in the human gastrointestinal tract, are promising candidates for the enzymatic treatment of celiac disease. Methods: Two wheat proteases were selected using AlphaFold2, produced in recombinant forms, and characterized. Their glutenase potentials under acidic or slightly acidic conditions were evaluated and compared with the properties of the previously characterized wheat glutenase Triticain-α. Results: All enzymes tested, Ta-P7, Ta-V6, and Triticain-α, were able to hydrolyze the model substrate (α-gliadin-derived epitope) in the pH range of 3.6–7.5. Nevertheless, Triticain-α performs the most efficient hydrolysis of the peptide substrate under the conditions of the gastrointestinal tract, according to its kinetic characteristics. In the wheat gluten degradation experiment at pH 4.6 and 37 °C, both Ta-P7 and Triticain-α cleaved the mixture almost completely within 5 min. In addition, Triticain-α and Ta-P7 significantly reduced the levels of toxic peptides compared to both intact gluten and gluten treated with pepsin-trypsin digestion as tested by the Ridascreen Gliadin Kit. Conclusions: Novel wheat proteases under investigation possess the expected glutenase activity to varying degrees; however, Triticain-α is a primary candidate for potential use in the enzymatic therapy of gluten-related disorders. Full article

(This article belongs to the Special Issue Plant-Based Bioactive Products for Pharmaceutical Applications)

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26 pages, 5853 KiB

Open AccessArticle

Kinin B₁ Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments

by Carolina Batista, João Victor Roza Cruz, Michele Siqueira, João Bosco Pesquero, Joice Stipursky and Fabio de Almeida Mendes

Pharmaceuticals 2025, 18(4), 591; https://doi.org/10.3390/ph18040591 - 18 Apr 2025

Abstract

Background/Objectives: The low permeability of the blood-brain barrier (BBB) represents a significant challenge to effective systemic chemotherapy for primary and metastatic brain cancers. Kinin receptors play a crucial role in modulating BBB permeability, and their agonist analogs have been explored in preclinical [...] Read more.

Background/Objectives: The low permeability of the blood-brain barrier (BBB) represents a significant challenge to effective systemic chemotherapy for primary and metastatic brain cancers. Kinin receptors play a crucial role in modulating BBB permeability, and their agonist analogs have been explored in preclinical animal models to enhance drug delivery to the brain. In this study, we investigated whether des-Arg⁹-bradykinin (DBK), a physiological agonist of kinin B₁ receptor (B1R), acts as a brain drug delivery adjuvant by promoting the transient opening of the BBB. Methods: Human brain microvascular endothelial cells (HBMECs) were treated with DBK in the culture medium and in conditioned media from glioblastoma cell lines, namely T98G (CMT98G) and U87MG (CMU87). Immunofluorescence, RT-qPCR, in-cell Western assay, and proximity ligation assay (PLA) were performed to analyze BBB components, kinin receptors and TLR4, a receptor associated with the kinin pathway and inflammation. The effect of DBK on enhancing paracellular molecule transport was evaluated using Evans blue dye (EB) quantification in a cell culture insert assay and in an in vivo model, where mice with and without brain tumors were treated with DBK. To assess the functional impact of the transient BBB opening induced by DBK, the chemotherapeutic drug doxorubicin (DOX) was administered. Results: Treatment with DBK facilitates the presence of EB in the brain parenchyma by transiently disrupting the BBB, as further evidenced by the increased paracellular passage of the dye in an in vitro assay. B1R activation by DBK induces transient BBB opening lasting less than 48 h, enhancing the bioavailability of the DOX within the brain parenchyma and glioma tumor mass. The interaction between B1R and TLR4 is disrupted by the secreted factors released by glioblastoma cells, as conditioned media from T98G and U87 reduce TLR4 staining in endothelial cells without affecting B1R expression. Conclusions: These results further support the potential of B1R activation as a strategy to enhance targeted drug delivery to the brain. Full article

(This article belongs to the Section Biopharmaceuticals)

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39 pages, 2165 KiB

Open AccessProject Report

Physicochemical Properties and Molecular Insights of Favipiravir and Roflumilast Solid Dispersions for COVID-19 Treatment

by Abdul Rauf and Saad Salman

Pharmaceuticals 2025, 18(4), 590; https://doi.org/10.3390/ph18040590 - 18 Apr 2025

Abstract

Background/Objectives: Fixed-dose combinations (FDCs) offer significant advantages for patients and healthcare systems by improving adherence and reducing pill burden. However, developing multi-drug formulations remains challenging due to complexities in drug compatibility, stability, and dissolution behavior. The COVID-19 pandemic has necessitated innovative therapeutic approaches. [...] Read more.

Background/Objectives: Fixed-dose combinations (FDCs) offer significant advantages for patients and healthcare systems by improving adherence and reducing pill burden. However, developing multi-drug formulations remains challenging due to complexities in drug compatibility, stability, and dissolution behavior. The COVID-19 pandemic has necessitated innovative therapeutic approaches. This study aims to develop and evaluate an FDC containing FR (an antiviral drug) and RT (a PDE4 inhibitor) for potential COVID-19 treatment. Methods: The proposed dual-layer FDC was formulated to achieve immediate release of RT using Klucel EXF and controlled release of FR using a combination of Klucel HXF and Compritol ATO888. Critical quality attributes, including drug–excipient compatibility, solid-state properties, tablet uniformity, and dissolution kinetics, were assessed. RT and FR quantification methods were developed and validated per international guidelines. Compatibility studies were conducted by combining excipients in fixed ratios with APIs, followed by stability testing. Results: No degradation or adverse interactions were observed between APIs and excipients. RT exhibited rapid dissolution within 30 min, while FR release was effectively controlled through a gel-forming matrix and lipid barrier. Bulk powder and tablet physical parameters met pharmacopeial standards, and content uniformity between layers was maintained. The formulation demonstrated a stable dissolution profile for both drugs, ensuring consistent drug release. Conclusions: The novel FDC of RT and FR exhibits favorable physicochemical properties, a stable dissolution profile, and potential for improved treatment efficacy in COVID-19 patients. By optimizing drug release mechanisms and ensuring formulation stability, this FDC could serve as a pharmaco-economically viable alternative to existing therapies, enhancing patient compliance and treatment outcomes. Full article

(This article belongs to the Section Pharmaceutical Technology)

30 pages, 16466 KiB

Open AccessReview

Natural Antifungal Alkaloids for Crop Protection: An Overview of the Latest Synthetic Approaches

by Denise Dozio, Francesca Sacchi, Andrea Pinto, Sabrina Dallavalle, Francesca Annunziata and Salvatore Princiotto

Pharmaceuticals 2025, 18(4), 589; https://doi.org/10.3390/ph18040589 - 18 Apr 2025

Abstract

Alkaloids are nitrogen-containing compounds naturally occurring in plants, microorganisms, and marine organisms. Potent biological activities have been reported to date, ranging from neuroprotective to antioxidant and anticancer effects. Alkaloids have recently gained attention as potential antifungal agents for crop protection due to their [...] Read more.

Alkaloids are nitrogen-containing compounds naturally occurring in plants, microorganisms, and marine organisms. Potent biological activities have been reported to date, ranging from neuroprotective to antioxidant and anticancer effects. Alkaloids have recently gained attention as potential antifungal agents for crop protection due to their broad spectrum of activity, eco-friendly nature, and ability to overcome some of the issues associated with synthetic fungicides, such as resistance development and environmental contamination. Several efforts have been made to obtain natural and nature-derived alkaloids endowed with significant activity against numerous pathogenic fungal strains. In this review, we collect synthetic strategies developed over the past decade to produce alkaloid fungicides for crop protection. Special emphasis is given to recent advancements in obtaining pure natural compounds and more potent analogs endowed with tailored and optimized properties. Full article

(This article belongs to the Special Issue Natural Products-Assisted Organic Synthesis in Medicinal Chemistry)

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17 pages, 294 KiB

Open AccessReview

Hormonal Treatment of Endometriosis: A Narrative Review

by Elvin Piriyev, Sven Schiermeier and Thomas Römer

Pharmaceuticals 2025, 18(4), 588; https://doi.org/10.3390/ph18040588 - 17 Apr 2025

Abstract

Background: Endometriosis is one of the most common gynecological diseases, affecting up to 10–15% of women of reproductive age. It is a chronic, estrogen-dependent condition that often presents with heterogeneous symptoms, complicating diagnosis and delaying treatment. Methods: This is a narrative [...] Read more.

Background: Endometriosis is one of the most common gynecological diseases, affecting up to 10–15% of women of reproductive age. It is a chronic, estrogen-dependent condition that often presents with heterogeneous symptoms, complicating diagnosis and delaying treatment. Methods: This is a narrative review based on a comprehensive analysis of recent literature regarding hormonal treatment options for endometriosis, including primary and adjuvant therapies. Results: Combined oral contraceptives (COCs) are effective in reducing dysmenorrhea, but show limited benefit for other symptoms and may not prevent disease progression. Progestins, particularly dienogest, demonstrate superior long-term efficacy with favorable side-effect profiles. GnRH agonists and antagonists are reserved for second-line treatment due to side effects and hypoestrogenism, but can significantly reduce endometriotic lesions. The levonorgestrel intrauterine system (LNG-IUS) is especially effective in patients with adenomyosis. Conclusions: Hormonal therapies are central to the management of endometriosis. Progestins are considered the most suitable long-term option. Despite promising results, evidence quality varies, and further studies are needed to establish long-term efficacy, patient-specific outcomes, and direct comparisons between agents. Full article

(This article belongs to the Special Issue Pharmacotherapy of Endometriosis)

23 pages, 3615 KiB

Open AccessArticle

Lipophilic Extracts of Portulaca oleracea L.: Analysis of Bioactive Fatty Acids Targeting Microbial and Cancer Pathways

by Dejan Stojković, Jelena Živković, Stefani Bolevich, Gokhan Zengin, Mehmet Veysi Cetiz, Sergey Bolevich and Marina Soković

Pharmaceuticals 2025, 18(4), 587; https://doi.org/10.3390/ph18040587 - 17 Apr 2025

Abstract

Background/Objectives: Portulaca oleracea L. (purslane) is a widely distributed plant known for its medicinal and nutritional properties. This study aims to evaluate the fatty acid composition and bioactivities of crude lipophilic extracts (chloroform/methanol 2:1) from purslane collected in Serbia and Greece, with [...] Read more.

Background/Objectives: Portulaca oleracea L. (purslane) is a widely distributed plant known for its medicinal and nutritional properties. This study aims to evaluate the fatty acid composition and bioactivities of crude lipophilic extracts (chloroform/methanol 2:1) from purslane collected in Serbia and Greece, with a focus on its antimicrobial and anticancer potential. Methods: Chemical analysis was conducted to determine the fatty acid composition of the extracts. Antibacterial activity was assessed using standard microdilution assays, while antibiofilm assays evaluated the extracts’ ability to inhibit biofilm formation. Cytotoxicity was tested on cancer cell lines (MCF7, HeLa, CaCo2, HepG2) and normal keratinocyte cells (HaCaT). Molecular docking and dynamics simulations were performed to explore the interactions of bioactive fatty acids with microbial and cancer-related proteins. Results: The analysis revealed significant levels of polyunsaturated fatty acids, with linoleic acid as the predominant fatty acid in both samples (31.42% and 34.51%). The Greek extract exhibited stronger antibacterial activity than the Serbian extract, particularly against Aspergillus versicolor, Pseudomonas aeruginosa, and Staphylococcus aureus. Antibiofilm assays showed up to 89.54% destruction at MIC levels, with notable reductions in exopolysaccharide and extracellular DNA production, especially for Greek samples. Cytotoxicity testing indicated moderate effects on cancer cell lines (IC₅₀ = 178.17–397.31 µg/mL) while being non-toxic to keratinocytes. Molecular docking identified strong interactions between key fatty acids and microbial and cancer-related proteins. Conclusions: These results highlight purslane’s potential as a source of bioactive compounds, particularly in antimicrobial and anticancer applications. The findings suggest that purslane extracts could be developed for therapeutic purposes targeting microbial infections and cancer. Full article

(This article belongs to the Special Issue Natural Pharmacons: Biologically Active Plant-Based Pharmaceuticals—4th Edition)

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24 pages, 1098 KiB

Open AccessArticle

Biomarker-Driven Pharmacokinetics and Efficacy of Polymyxin B in Critically Ill Patients with XDR-GN Pneumonia

by Wei Zuo, Qianlin Wang, Longxiang Su, Jiaxin Yu, Hongwei Fan, Qiang Fu, Yun Long and Bo Zhang

Pharmaceuticals 2025, 18(4), 586; https://doi.org/10.3390/ph18040586 - 17 Apr 2025

Abstract

Background: Achieving pharmacokinetic/pharmacodynamic (PK/PD) targets is critical for improving treatment success, particularly in critically ill patients. This study investigates the role of inflammatory biomarkers and their influence on the PK/PD characteristics of polymyxin B (PMB) in patients with extensively drug-resistant Gram-negative (XDR-GN) bacterial [...] Read more.

Background: Achieving pharmacokinetic/pharmacodynamic (PK/PD) targets is critical for improving treatment success, particularly in critically ill patients. This study investigates the role of inflammatory biomarkers and their influence on the PK/PD characteristics of polymyxin B (PMB) in patients with extensively drug-resistant Gram-negative (XDR-GN) bacterial nosocomial pneumonia. Methods: Serial blood and/or bronchoalveolar lavage fluid (BALF) samples were collected at specified time points and analyzed for PMB and/or inflammatory biomarkers, including IL-6 and IL-10. Clinical data were also recorded, and their correlations with PK parameters were further analyzed. Results: Among the 27 enrolled patients, 22 (81.5%) achieved treatment success. The pharmacokinetic parameters of PMB included a maximum plasma concentration (C_max) of 8.3 µg/mL, clearance (CL) of 1.55 L/h, volume of distribution (Vd) of 30.44 L, half-life (t_1/2) of 19.56 h, steady-state area under the plasma concentration–time curve from time 0 to 24 h (AUC_ss,0–24h) of 110.08 h·µg/mL, and a plasma protein-binding ratio of 85.53%. The AUC_ss,0–24h metric was identified as a robust predictor of clinical efficacy, with an optimal cutoff value of 77.27 h·µg/mL. Notably, 48.15% of patients achieved the target AUC_ss,0–24h range of 50–100 h·µg/mL, with 76.95% of these patients attaining treatment success. Another 48.15% of patients exceeded this target, and 92.31% of this subgroup achieved treatment success. PMB demonstrated limited pulmonary penetration, with an epithelial lining fluid (ELF)/plasma ratio of 15.69% [16.86, 18.15]. Furthermore, TNF-α and the IL-6/IL-10 ratio were significantly correlated with PMB PK parameters. Conclusions: Our and others’ studies suggest heterogeneity of PMB PK parameters in critically ill patients. The majority of critically ill patients achieved or surpassed the recommended PK/PD targets and attained treatment success through intravenous administration of PMB at a simplified fixed dose. However, PMB did not achieve satisfactory pulmonary concentrations, suggesting that its efficacy may involve alternative mechanisms. The modulation of inflammatory responses may play a pivotal role in the treatment of severe infections, highlighting the potential for biomarker-guided therapeutic strategies. Full article

(This article belongs to the Section Pharmacology)

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21 pages, 605 KiB

Open AccessReview

Synergizing Success: The Role of Anlotinib Combinations in Advanced Non-Small Cell Lung Cancer Treatment

by Helal F. Hetta, Hashim M. Aljohani, Nizar Sirag, Hassabelrasoul Elfadil, Ayman Salama, Rand Al-Twalhy, Danah Alanazi, Manal D. Al-johani, Jumanah H. Albalawi, Rinad M. Al-Otaibi, Raghad A. Alsharif and Reem Sayad

Pharmaceuticals 2025, 18(4), 585; https://doi.org/10.3390/ph18040585 - 16 Apr 2025

Abstract

Anlotinib, a novel receptor tyrosine kinase inhibitor that is taken orally, targets several RTKs and is authorized as a third-line treatment for patients with advanced non-small cell lung cancer (NSCLC). Anlotinib is also used in combination with immunotherapy or chemotherapy for advanced NSCLC. [...] Read more.

Anlotinib, a novel receptor tyrosine kinase inhibitor that is taken orally, targets several RTKs and is authorized as a third-line treatment for patients with advanced non-small cell lung cancer (NSCLC). Anlotinib is also used in combination with immunotherapy or chemotherapy for advanced NSCLC. We aimed to explore the efficacy and safety of anlotinib-based regimens in NSCLC treatment, focusing on combination therapies. We also addressed challenges that hinder oncologists from using it, such as toxicity and resistance mechanisms. A systematic approach involves searching the National Institute of Health PubMed, Scopus, MedLine, and Web of Science databases up to April 2024. Relevant studies were identified and analyzed for their methodologies, outcomes, and patient characteristics. Findings revealed that numerous effective combination regimens, such as anlotinib with platinum-based chemotherapy and anlotinib combined with PD-1 blockades, have shown positive results in terms of progression-free survival (PFS), overall survival (OS), and objective response rate (ORR). On the other hand, NSCLC treatment faces hurdles due to drug resistance and its toxicity profile. These challenges underscore the need for continued research and optimization of treatment strategies. Full article

(This article belongs to the Special Issue Cancer Chemoradiotherapy)

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14 pages, 12806 KiB

Open AccessArticle

Evaluation of Chitosan-Based Axiostat as Hemostatic Dressing for Endovascular Procedures in Patients with Leriche Syndrome on Anticoagulant Therapy

by Paolo Perri, Federica Curcio, Michele De Luca, Paolo Piro, Sonia Trombino and Roberta Cassano

Pharmaceuticals 2025, 18(4), 584; https://doi.org/10.3390/ph18040584 - 16 Apr 2025

Abstract

Background/Objectives: The safe completion of a non-invasive procedure is crucial to the success of an endovascular approach. Chitosan, a natural polysaccharide derived from chitin, is an ideal material for the study and application of medical devices in post-operative wound management. Methods: The present [...] Read more.

Background/Objectives: The safe completion of a non-invasive procedure is crucial to the success of an endovascular approach. Chitosan, a natural polysaccharide derived from chitin, is an ideal material for the study and application of medical devices in post-operative wound management. Methods: The present work is based on a retrospective study conducted on a sample of patients treated with Axiostat (a sterile, single-use, non-absorbable dressing), composed of 100% chitosan and designed to instantly stop bleeding through a mucus adhesion mechanism for the treatment of conditions such as Leriche’s syndrome. The objective was to evaluate the efficacy and safety of the hemostatic Axiostat dressing in patients undergoing anticoagulant and/or antiplatelet therapy in whom endovascular procedures using the axillary artery as an access site are performed to treat Leriche syndrome. Results: The obtained results showed that Axiostat is safe and effective in promoting hemostasis at the axillary vascular access site even when prolonged hemostasis was required in patients on antiplatelet and anticoagulant therapy. The mean time to hemostasis was 5.75 min in all types of patients considered. Full article

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20 pages, 2359 KiB

Open AccessArticle

Prognostic Factors and Talaporfin Sodium Concentration in Photodynamic Therapy for Recurrent Grade 4 Glioma

by Mikoto Onodera, Shuji Kitahara, Yasuto Sato, Takakazu Kawamata, Yoshihiro Muragaki and Ken Masamune

Pharmaceuticals 2025, 18(4), 583; https://doi.org/10.3390/ph18040583 - 16 Apr 2025

Abstract

Background: Although extensive resection improves the prognosis of gliomas, it risks impairing critical brain functions. Photodynamic therapy (PDT) utilizing talaporfin sodium (TS) targets tumor cells upon light activation. Despite its approval in Japan, TS application remains restricted, and factors influencing its efficacy are [...] Read more.

Background: Although extensive resection improves the prognosis of gliomas, it risks impairing critical brain functions. Photodynamic therapy (PDT) utilizing talaporfin sodium (TS) targets tumor cells upon light activation. Despite its approval in Japan, TS application remains restricted, and factors influencing its efficacy are unclear. We aimed to identify TS efficacy determinants to optimize treatment outcomes. Methods: Data from 171 patients with grade 4 glioma who underwent surgery and PDT at Tokyo Women’s Medical University Hospital between January 2017 and March 2024 were retrospectively analyzed. Clinical variables evaluated included age, sex, genotype, Karnofsky Performance Status (KPS), serum albumin (Alb) levels, MIB-1 expression levels, and medication history. TS concentrations in tumor tissues were quantitatively assessed in 82 patients (41 primary, 41 recurrent). Survival outcomes were analyzed. RNA-seq was performed on the three highest and three lowest TS concentration samples with significant TS concentration variations to investigate corresponding gene expression changes. Results: Multivariate analysis identified KPS (hazard ratio [95% confidence interval]: 0.96 [0.93–0.99], p = 0.01) and Alb (3.68 [1.05–13.76], p = 0.047) as independent prognostic factors. In recurrent cases, higher TS concentrations were significantly associated with improved survival (p = 0.0454). RNA-seq analysis indicated decreased expression of ACTB and PDPN genes in samples with lower TS concentrations, suggesting potential resistance mechanisms. Conclusions: TS concentration is a critical determinant of PDT efficacy, especially in recurrent glioma, highlighting its prognostic significance. Alb may affect treatment outcomes by mediating TS binding. RNA-seq findings imply that low TS concentrations may suppress immune and stress response-related genes, potentially diminishing PDT sensitivity. Full article

(This article belongs to the Special Issue New Platforms for Cancer Treatment—Emerging Advances)

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11 pages, 1225 KiB

Open AccessSystematic Review

Exploring the Pharmacokinetics of Drugs in Disabled Saudi Patients: A Systematic Review

by Faleh Alqahtani, Saeed A. Al Awadh and Muhammad Fawad Rasool

Pharmaceuticals 2025, 18(4), 582; https://doi.org/10.3390/ph18040582 - 16 Apr 2025

Abstract

Background/Objectives: Disability is a term that involves mental, intellectual, or sensory impairment resulting in the loss of one’s ability to walk or perform the activities necessary to live in a society. This study aims to collect all the data regarding the absorption, [...] Read more.

Background/Objectives: Disability is a term that involves mental, intellectual, or sensory impairment resulting in the loss of one’s ability to walk or perform the activities necessary to live in a society. This study aims to collect all the data regarding the absorption, distribution, and disposition of drugs in disabled Saudi patients, i.e., patients suffering from epilepsy, cancer, cardiovascular diseases, etc., and then compare these results with data reported in other ethnicities. Methods: An exhaustive online search used the key terms in Google Scholar, PubMed, Cochrane Library, and Science Direct to extract all articles that met the eligibility criteria. All research studies containing pharmacokinetic (PK) parameters (area under the curve from 0 to infinity (AUC_0–∞), maximal plasma concentration (C_max), clearance (CL), volume of distribution, time to reach maximum plasma concentration, and half-life) were included in this review. Results: In pediatric epileptic patients, carbamazepine showed a notable decrease in C_max with increasing age, which may be due to ontogenetic changes in its disposition. The AUC_0–∞ of busulphan in adult hematopoietic stem cell transplantation patients was recorded as 4392.5 ± 1354.65 μg·h/mL, with high inter-individual variability. Moreover, the CL of vancomycin was reported to be 25% higher among cancer patients in comparison to non-cancer subjects. Conclusions: The complications in disabled patients due to alterations in cytochrome P450 enzymes, pathophysiology, genetics, and ethnicity emphasize the significance of patient-centered drug dosing. These findings may aid healthcare physicians in refining therapeutic care in this population. Full article

(This article belongs to the Special Issue Population Pharmacokinetics and Pharmacogenetics)

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16 pages, 1742 KiB

Open AccessArticle

Binding Affinity of Synthetic Cannabinoids to Human Serum Albumin: Site Characterization and Interaction Insights

by Rita M. G. Santos, Rita Lima, Sara Cravo, Pedro Alexandrino Fernandes, Fernando Remião and Carla Fernandes

Pharmaceuticals 2025, 18(4), 581; https://doi.org/10.3390/ph18040581 - 16 Apr 2025

Abstract

Background/Objectives: High-performance affinity chromatography (HPAC) was used to investigate the binding affinity of a series of synthetic cannabinoids, a widely abused class of new psychoactive substances, to human serum albumin (HSA) and obtain insights into the binding sites. To better understand the recognition [...] Read more.

Background/Objectives: High-performance affinity chromatography (HPAC) was used to investigate the binding affinity of a series of synthetic cannabinoids, a widely abused class of new psychoactive substances, to human serum albumin (HSA) and obtain insights into the binding sites. To better understand the recognition mechanisms, molecular docking studies were conducted. Methods: Binding affinity was assessed through zonal elution approach Additionally, displacement chromatography with site-specific probes provided insights into the HSA binding sites of five synthetic cannabinoids. Results: That these drugs exhibit extensive binding to HSA, with values ranging from 98.7% to 99.9%. Competition for site I was observed between warfarin and four synthetic cannabinoids (5F-AMB, AB-PINACA, AMB-FUBINACA, and AB-CHMINACA). Furthermore, AB-CHMINACA also competed with L-tryptophan for site II. The binding affinity of all synthetic cannabinoids increased in the presence of (S)-ibuprofen. Molecular docking studies supported the experimental findings, reinforcing the insights gained. Conclusions: The key novelty of this study lies in analyzing, for the first time, the binding affinity of synthetic cannabinoids to HSA through HPAC and molecular docking. These results may improve our understanding of their toxicokinetic behavior and help in predicting possible competitive interactions that could influence HSA binding and, consequently, their activity and toxicity. This study is the first to describe the binding affinity of synthetic cannabinoids to HSA, elucidate their recognition mechanisms, identify binding sites, and characterize their interactions with the protein. Full article

(This article belongs to the Special Issue Innovative Tools for Drug Analysis and Therapeutic Drug Monitoring (TDM))

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25 pages, 7873 KiB

Open AccessReview

Recent Developments of 1,3,4-Thiadiazole Compounds as Anticancer Agents

by Serena Indelicato, David Bongiorno, Manuela Mauro and Stella Cascioferro

Pharmaceuticals 2025, 18(4), 580; https://doi.org/10.3390/ph18040580 - 16 Apr 2025

Abstract

The World Health Organization has recently underlined the increasing global burden of cancer, with a particularly alarming impact on underserved populations. In recent years, 1,3,4-thiadiazole has emerged as a versatile pharmacophore to obtain bioactive compounds. The pharmacological properties of this ring are primarily [...] Read more.

The World Health Organization has recently underlined the increasing global burden of cancer, with a particularly alarming impact on underserved populations. In recent years, 1,3,4-thiadiazole has emerged as a versatile pharmacophore to obtain bioactive compounds. The pharmacological properties of this ring are primarily attributed to its role as a bioisostere of pyrimidine, the core structure of three nucleic bases. This structural feature endows 1,3,4-thiadiazole derivatives with the ability to interfere with DNA replication processes. Additionally, the mesoionic behavior of this heterocycle gives it important properties, such as the ability to cross biological membranes and interact with target proteins. Noteworthy, in analogy to the other sulfur heterocycles, the presence of C-S σ* orbitals, conferring small regions of low electron density on the sulfur atom, makes interaction with the target easier. This review focuses on the most promising anticancer agents with 1,3,4-thiadiazole structure reported in the past five years, providing information that may be useful to medicinal chemists who intend to develop new anticancer derivatives. Full article

(This article belongs to the Special Issue Drug Target Exploration and Drug Design & Development Based on Small Molecule)

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18 pages, 3144 KiB

Open AccessArticle

Comparative Forced Degradation Study of Anticomplement C5 Biosimilar and Originator Monoclonal Antibodies

by Merve Celik Yamaci, Ceren Pamukcu, Yigit Erdemgil, Ahmet Emin Atik, Zeynep Zulfiye Yildirim Keles and Ozge Can

Pharmaceuticals 2025, 18(4), 579; https://doi.org/10.3390/ph18040579 - 16 Apr 2025

Abstract

Background/Objectives: The stress testing of biotherapeutic products is a critical component of drug development, enabling the assessment of stability, biosimilarity, and degradation pathways. Subjecting biosimilar monoclonal antibodies to controlled stress conditions yields essential insights into their structural and functional integrity, informing formulation [...] Read more.

Background/Objectives: The stress testing of biotherapeutic products is a critical component of drug development, enabling the assessment of stability, biosimilarity, and degradation pathways. Subjecting biosimilar monoclonal antibodies to controlled stress conditions yields essential insights into their structural and functional integrity, informing formulation optimization and mitigating risks before clinical trials. In this study, biosimilar products were comprehensively characterized and compared with originator products under forced degradation. The aim was to expose the products to different stress conditions such as oxidative, pH, thermal, freeze/thaw, and agitation. The products were then tested at defined time points using validated analytical methods. Methods: This study employed size-exclusion chromatography to detect aggregated forms. Isoelectric focusing characterized protein charge variants (e.g., acidic/basic isoforms) from post-translational modifications, while capillary electrophoresis quantified product-related impurities (aggregates and fragments). In addition, a complement assay was used to determine the efficacy and potency under specific stress conditions. Results: Our findings showed that biosimilar and originator products exhibited similar degradation profiles. The biosimilar monoclonal antibody was found to be analytically similar to the originator product in terms of critical parameters related to efficacy and safety under various stress conditions such as aggregation profile, biological activity, and charge variant distribution. Conclusions: Forced degradation studies facilitated the comprehensive and well-validated characterization of the structure and biological activity of biosimilar monoclonal antibody products. Full article

(This article belongs to the Special Issue Biosimilars Development Strategies)

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17 pages, 1677 KiB

Open AccessArticle

Phytochemical Profiling and Biological Evaluation of Dianthus sylvestris subsp. aristidis: A Chromatographic and Mass Spectrometry Approach to Uncovering Bioactive Metabolites for Dermatological and Metabolic Disorder Management

by Amina Bouzana, Zohra Chekroud, Imène Becheker, Fatima Kamah, Nora Sakhraoui, Chawki Bensouici, Fehmi Boufahja, Sulaiman A. Alsalamah, Mohammed I. Alghonaim, Stefania Garzoli and Hamdi Bendif

Pharmaceuticals 2025, 18(4), 578; https://doi.org/10.3390/ph18040578 - 16 Apr 2025

Abstract

Background/Objectives: This study provides the first comprehensive phytochemical composition and biological evaluation of Dianthus sylvestris subsp. aristidis (Batt.) Greuter & Burdet, a plant endemic to Algeria with unexplored pharmacological potential. The objective is to identify novel bioactive metabolites in the plant’s extracts [...] Read more.

Background/Objectives: This study provides the first comprehensive phytochemical composition and biological evaluation of Dianthus sylvestris subsp. aristidis (Batt.) Greuter & Burdet, a plant endemic to Algeria with unexplored pharmacological potential. The objective is to identify novel bioactive metabolites in the plant’s extracts and assess their potential applications for skincare and metabolic disorder management, addressing gaps in the current understanding of its medicinal value. Methods: Liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) profiling was used to analyze the hydromethanolic (HMeOH) leaf extract and identify bioactive compounds. The biological activities of HMeOH, ethyl acetate (EtOAc), and butanolic (n-BuOH) extracts were tested for cytotoxicity using the brine shrimp lethality test, photoprotective potential by calculating the sun protection factor (SPF), and enzymatic inhibitory activities against alpha-amylase, urease, and tyrosinase. Results: The LC-ESI-MS/MS profiling of the MeOH extract identified 22 bioactive compounds, including phenolic acids and flavonoids, some of which have not been previously reported in this species. Cytotoxicity tests showed that all extracts were non-toxic (half-lethal concentration (LC₅₀) > 100 micrograms per milliliter). The SPF values indicated significant photoprotective potential, with EtOAc (SPF = 45.19 ± 0.73) and n-BuOH (SPF = 43.81 ± 0.59) extracts showing high sun protection activity. The n-BuOH extract exhibited strong alpha-amylase inhibitory activity (half-maximal inhibitory concentration (IC₅₀) = 307.08 micrograms per milliliter), surpassing the standard acarbose (IC₅₀ = 3650.93 micrograms per milliliter), suggesting potential applications in diabetes management. Conclusions: Dianthus sylvestris subsp. aristidis demonstrates significant pharmacological potential as a source of bioactive secondary metabolites for skincare and metabolic disorder management. These findings provide new insights into the plant’s therapeutic potential and set a foundation for future pharmacological and clinical investigations. Full article

(This article belongs to the Section Natural Products)

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35 pages, 7644 KiB

Open AccessReview

Drug Discovery for Histone Deacetylase Inhibition: Past, Present and Future of Zinc-Binding Groups

by Gustavo Salgado Pires, Heber Victor Tolomeu, Daniel Alencar Rodrigues, Lídia Moreira Lima, Carlos Alberto Manssour Fraga and Pedro de Sena Murteira Pinheiro

Pharmaceuticals 2025, 18(4), 577; https://doi.org/10.3390/ph18040577 - 16 Apr 2025

Abstract

Histone deacetylases (HDACs) are key regulators of gene expression, influencing chromatin remodeling and playing a crucial role in various physiological and pathological processes. Aberrant HDAC activity has been linked to cancer, neurodegenerative disorders, and inflammatory diseases, making these enzymes attractive therapeutic targets. HDAC [...] Read more.

Histone deacetylases (HDACs) are key regulators of gene expression, influencing chromatin remodeling and playing a crucial role in various physiological and pathological processes. Aberrant HDAC activity has been linked to cancer, neurodegenerative disorders, and inflammatory diseases, making these enzymes attractive therapeutic targets. HDAC inhibitors (HDACis) have gained significant attention, particularly those containing zinc-binding groups (ZBGs), which interact directly with the catalytic zinc ion in the enzyme’s active site. The structural diversity of ZBGs profoundly impacts the potency, selectivity, and pharmacokinetics of HDACis. While hydroxamic acids remain the most widely used ZBGs, their limitations, such as metabolic instability and off-target effects, have driven the development of alternative scaffolds, including ortho-aminoanilides, mercaptoacetamides, alkylhydrazides, oxadiazoles, and more. This review explores the structural and mechanistic aspects of different ZBGs, their interactions with HDAC isoforms, and their influence on inhibitor selectivity. Advances in structure-based drug design have allowed the fine-tuning of HDACi pharmacophores, leading to more selective and efficacious compounds with improved drug-like properties. Understanding the nuances of ZBG interactions is essential for the rational design of next-generation HDACis, with potential applications in oncology, neuroprotection, and immunotherapy. Full article

(This article belongs to the Special Issue A Commemorative Special Issue in Honor of Professor Carlos Alberto Manssour Fraga: Medicinal Chemistry’s Efforts to Discover Novel Bioactive Molecules for Complex Diseases)

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24 pages, 2317 KiB

Open AccessArticle

Bioactivities and Chemotaxonomy of Four Heracleum Species: A Comparative Study Across Plant Parts

by Tugce Ince Kose, Gamze Benli Yardimci, Damla Kirci, Derya Cicek Polat, Betul Demirci, Mujde Eryilmaz and Ceyda Sibel Kilic

Pharmaceuticals 2025, 18(4), 576; https://doi.org/10.3390/ph18040576 - 16 Apr 2025

Abstract

Background/Objectives: This study investigates the phytochemical profile, essential oil composition, and bioactivities—including antioxidant, antimicrobial, antibio-film, and anti-quorum sensing (QS) activities—of four Heracleum L. species (H. crenatifolium Boiss, H. paphlagonicum Czeczott, H. sphondylium subsp. montanum Schleich. ex Gaudin, and H. pastinacifolium subsp. [...] Read more.

Background/Objectives: This study investigates the phytochemical profile, essential oil composition, and bioactivities—including antioxidant, antimicrobial, antibio-film, and anti-quorum sensing (QS) activities—of four Heracleum L. species (H. crenatifolium Boiss, H. paphlagonicum Czeczott, H. sphondylium subsp. montanum Schleich. ex Gaudin, and H. pastinacifolium subsp. incanum (Boiss. & A.Huet) P.H.Davis). Methods: Total phenolic and flavonoid contents were quantified using the Folin–Ciocalteu and aluminum chloride colorimetric methods, respectively. Essential oils were extracted by hydrodistillation and analyzed via Gas Chromatography–Flame Ionization Detector (GC–FID) and Gas Chromatography–Mass Spectrometry (GC–MS), while Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) evaluated chemical variability among the species. Antioxidant activities were assessed using DPPH and ABTS free radical scavenging assays. Antimicrobial activity was assessed using the broth microdilution method to determine Minimum Inhibitory Concentration (MIC) values, while antibiofilm activity was evaluated using an in vitro microplate-based biofilm model against Pseudomonas aeruginosa PAO1. Anti-QS activity was analyzed using a disc diffusion assay with Chromobacterium violaceum ATCC 12472 as the reporter strain. Results: It was observed that the amounts of total phenolic compounds and total flavonoids were higher in root extracts than in aerial parts extracts for the four species in this study (H. sphondylium subsp. montanum excluding phenolic content). In the analysis of essential oil, it was determined that the major component in the roots was mostly myristicin, and in the fruits it was mostly octyl acetate. Phenolic and flavonoid contents were positively correlated with antioxidant activity. Methanol and n-hexane extracts of H. pastinacifolium (aerial parts) and n-hexane extracts of H. paphlagonicum (root) exhibited notable antimicrobial activity, primarily against Gram-positive bacteria, but none of the extracts showed activity against Klebsiella pneumoniae ATCC 13383 or P. aeruginosa ATCC 27853. Among methanol extracts, H. pastinacifolium (aerial parts) exhibited the highest antibiofilm activity (73.2%), while H. paphlagonicum (aerial parts) showed the highest activity among n-hexane extracts (75.5%). All n-hexane extracts exhibited anti-QS activity, whereas the methanol extracts showed no activity. Conclusions: These findings underscore the chemical diversity and bioactive potential of Heracleum species, contributing to the chemotaxonomic understanding of the genus and supporting their potential applications in medicine and industry. To our knowledge, this is the first study that reveals the antibiofilm and anti-QS properties of these Heracleum species. Full article

(This article belongs to the Special Issue Natural Products and Their Modifications as Agents Used in the Fight against Pathogens)

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16 pages, 3371 KiB

Open AccessArticle

A Prospective Randomized Pilot Study on the Efficacy of a Dietary Supplementation Regimen of Vitamin E and Selenium for the Prevention of Fluoroquinolone-Induced Tendinopathy

by Oana-Maria Mișcă, Liviu-Coriolan Mișcă, Bogdan Huzum, Andreea-Adriana Neamţu, Simona Cerbu, Daniel-Raul Chioibaș, Petrișor Zorin Crăiniceanu and Andrei Gheorghe Marius Motoc

Pharmaceuticals 2025, 18(4), 575; https://doi.org/10.3390/ph18040575 - 15 Apr 2025

Abstract

Background: Fluoroquinolone-induced tendinopathy is a clinically significant adverse effect associated with this class of antibiotics, particularly affecting the Achilles tendon. Despite its growing recognition, the precise pathophysiological mechanisms remain incompletely understood, with hypotheses referencing increased matrix metalloproteinase activity, collagen degradation, and oxidative [...] Read more.

Background: Fluoroquinolone-induced tendinopathy is a clinically significant adverse effect associated with this class of antibiotics, particularly affecting the Achilles tendon. Despite its growing recognition, the precise pathophysiological mechanisms remain incompletely understood, with hypotheses referencing increased matrix metalloproteinase activity, collagen degradation, and oxidative stress. Methods: This prospective randomized pilot study evaluates the potential protective effectiveness of vitamin E and selenium supplementation in mitigating fluoroquinolone-induced tendinopathy. The study was conducted on 25 patients receiving 500 mg/day levofloxacin antibiotherapy, randomly divided into a control group and an experimental group—vitamin E (400 IU/day) and selenium (200 µg/day), oral supplementation for 28 days. Clinical assessment of the pain level through the VAS score and of functionality through the VISA-A score was performed, alongside ultrasound imaging of the Achilles tendon. To assess potential toxicity and ensure adherence to the supplementation protocol, serial biochemical analyses of serum vitamin E and selenium were performed at predetermined intervals. Results: A significant improvement was observed in pain scores (p = 0.0120) and functional outcomes (p = 0.0340) when comparing the control and experimental groups at the three-month follow-up. Ultrasound analysis revealed reduced tendon thickness and neovascularization, supporting structural recovery. Although the incidence of tendinopathy was lower in the interventional group (13.3% vs. 40%), statistical significance was not reached, possibly due to the small sample size. Conclusions: These findings suggest that antioxidant supplementation with vitamin E and selenium may provide a protective effect against fluoroquinolone-induced tendinopathy, warranting further investigation in larger randomized clinical trials. Full article

(This article belongs to the Special Issue Fluoroquinolones)

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26 pages, 5850 KiB

Open AccessArticle

Lipid-Based Nanoformulations of [6]-Gingerol for the Chemoprevention of Benzo[a] Pyrene-Induced Lung Carcinogenesis: Preclinical Evidence

by Faris Alrumaihi, Ali Yousif Babiker and Arif Khan

Pharmaceuticals 2025, 18(4), 574; https://doi.org/10.3390/ph18040574 - 15 Apr 2025

Abstract

Background/Objectives: [6]-Gingerol ([6]-G), a bioactive compound derived from Zingiber officinale (ginger), exhibits strong anticancer potential but is hindered by poor aqueous solubility and low bioavailability. This study aimed to develop and evaluate PEGylated liposomal [6]-G (6-G-Lip) to enhance its stability, bioavailability, and chemopreventive [...] Read more.

Background/Objectives: [6]-Gingerol ([6]-G), a bioactive compound derived from Zingiber officinale (ginger), exhibits strong anticancer potential but is hindered by poor aqueous solubility and low bioavailability. This study aimed to develop and evaluate PEGylated liposomal [6]-G (6-G-Lip) to enhance its stability, bioavailability, and chemopreventive efficacy in benzo[a]pyrene (BaP)-induced lung carcinogenesis. Methods: 6-G-Lip was synthesized using a modified thin-film hydration technique and characterized for size, polydispersity index (PDI), zeta potential, encapsulation efficiency (EE%), and release kinetics. The chemopreventive effects were assessed in BaP-induced lung cancer in Swiss albino mice, with prophylactic 6-G-Lip administration from two weeks before BaP exposure through 21 weeks. Cancer biomarkers, antioxidant enzyme activity, reactive oxygen species (ROS) generation, induction of apoptosis, and histopathological alterations were analyzed. Results: 6-G-Lip exhibited a particle size of 129.7 nm, a polydispersity index (PDI) of 0.16, a zeta potential of −18.2 mV, and an encapsulation efficiency (EE%) of 91%, ensuring stability and effective drug loading. The formulation exhibited a controlled release profile, with 26.5% and 47.5% of [6]-G released in PBS and serum, respectively, at 72 h. 6-G-Lip significantly lowered cancer biomarkers, restored antioxidant defenses (SOD: 5.60 U/min/mg protein; CAT: 166.66 μm H₂O₂/min/mg protein), reduced lipid peroxidation (MDA: 3.3 nm/min/mg protein), and induced apoptosis (42.2%), highlighting its chemopreventive efficacy. Conclusions: This study is the first to prepare, characterize, and evaluate PEGylated [6]-G-Lip for the chemoprevention of lung cancer. It modulates oxidative stress, restores biochemical homeostasis, and selectively induces apoptosis. These findings support 6-G-Lip as a promising nanotherapeutic strategy for cancer prevention. Full article

(This article belongs to the Special Issue Plant-Derived Nanotherapeutics and Nanocarriers: Recent Progress and Future Directions)

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