Pharmaceuticals

19 pages, 1070 KiB

Open AccessArticle

Sparstolonin B Suppresses Proliferation and Modulates Toll-like Receptor Signaling and Inflammatory Pathways in Human Colorectal Cancer Cells

by Bürke Çırçırlı, Çağatay Yılmaz, Tuğçe Çeker, Zerrin Barut, Esma Kırımlıoğlu and Mutay Aslan

Pharmaceuticals 2025, 18(3), 300; https://doi.org/10.3390/ph18030300 - 21 Feb 2025

Abstract

Background: Sparstolonin B (SsnB), a natural compound with anti-inflammatory and anti-proliferative properties, was investigated for its effects on cell viability, apoptosis, and inflammatory pathways in human colorectal cancer cells (HCT-116) and healthy human fibroblasts (BJ). Phorbol 12-myristate 13-acetate (PMA), a tumor promoter and [...] Read more.

Background: Sparstolonin B (SsnB), a natural compound with anti-inflammatory and anti-proliferative properties, was investigated for its effects on cell viability, apoptosis, and inflammatory pathways in human colorectal cancer cells (HCT-116) and healthy human fibroblasts (BJ). Phorbol 12-myristate 13-acetate (PMA), a tumor promoter and inflammatory activator, was used to stimulate proliferation and inflammatory pathways. Methods: HCT-116 and BJ cells were treated with SsnB (3.125–50 μM) or PMA (1–10 nM) for 12–18 h. Cell viability was assessed using MTT analysis, while apoptosis was evaluated through cleaved caspase-3 staining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and flow cytometry. Proliferation was analyzed through proliferating cell nuclear antigen (PCNA) staining. Toll-like receptor (TLR) signaling, cytokine expression, and sphingolipid levels were measured using immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and mass spectrometry, respectively. Results: SsnB reduced HCT-116 cell viability in a dose- and time-dependent manner with minimal effects on BJ cells. SsnB (25 μM, 12 h) decreased HCT-116 viability 0.6-fold, while PMA (10 nM, 12 h) increased it 2-fold (p < 0.01). No significant change was observed in BJ cells. PCNA fluorescence staining increased 2-fold with PMA and decreased 0.4-fold with SsnB (p < 0.001). PMA upregulated TLR2 and TLR4 mRNA and protein levels, with MyD88, p-ERK, and pNF-κB fluorescence increasing 2.1-, 1.5-, and 1.7-fold, respectively (p < 0.001). PMA elevated TNF-α, IL-1β, and IL-6 levels (p < 0.01). SsnB suppressed PMA-induced effects and promoted apoptosis, increasing cleaved caspase-3 levels by 1.5-fold and TUNEL staining by 1.9-fold (p < 0.01). Flow cytometry confirmed a significant increase in early and late apoptotic cells in the SsnB group. SsnB also increased ceramide (C18, C20, C22, and C24) levels (1.3- to 2.5-fold, p < 0.01) while reducing PMA-induced S1P and C1P increases (p < 0.01). Conclusions: SsnB selectively inhibits proliferation, induces apoptosis, and modulates inflammatory and sphingolipid pathways in colorectal cancer cells, with minimal toxicity to healthy fibroblasts, supporting its potential as a targeted therapeutic agent. Full article

(This article belongs to the Section Natural Products)

24 pages, 3256 KiB

Open AccessArticle

Supercritical Extraction and Identification of Bioactive Compounds in Dryopteris fragrans (L.) Schott

by Mayya P. Razgonova, Zhanna M. Okhlopkova, Muhammad A. Nawaz, Polina S. Egorova and Kirill S. Golokhvast

Pharmaceuticals 2025, 18(3), 299; https://doi.org/10.3390/ph18030299 - 21 Feb 2025

Abstract

Background: This is a comparative metabolomic study of the medicinal plant Dryopteris fragrans (L.) Schott from the family Dryopteridaceae Herter (or Aspidiaceae Mett. ex Frank) growing under cold pole conditions in the Oymyakon region of the Republic of Sakha (Yakutia). Methods: The aerial [...] Read more.

Background: This is a comparative metabolomic study of the medicinal plant Dryopteris fragrans (L.) Schott from the family Dryopteridaceae Herter (or Aspidiaceae Mett. ex Frank) growing under cold pole conditions in the Oymyakon region of the Republic of Sakha (Yakutia). Methods: The aerial parts of D. fragrans were subjected to extraction using supercritical CO₂ extraction and maceration methods. Several experimental conditions were investigated, including a pressure range of 50–300 bar and a temperature range of 31–60 °C. A 1% volume of ethanol was used as a co-solvent in the liquid phase of the extraction. Results: The most effective D. fragrans extraction conditions were 200 Bar pressure and a temperature of 55 °C. Tandem mass spectrometry was used to detect the target analytes. A total of 141 bioactive compounds (86 compounds from the polyphenol group and 55 compounds from other chemical groups) were tentatively identified in extracts of aerial parts of D. fragrans. Among these, thirty chemical constituents from the polyphenol group were identified for the first time. Other compound classes that were newly identified in D. fragrans include naphthoquinones (5,8-dihydroxy-6-methyl-2,3-dihydro-1,4-naphthoquinone, 1,8-dihydroxy-anthraquinone, 1,4,8-trihydroxyanthraquinone, chrysophanol, etc.), diterpenoids (tanshinone IIa, cryptotanshinone, isocryptotanshinone II, tanshinone IIb, etc.), polysaccharides, triterpenoids, and sesquiterpenes. Conclusions: These results highlight that D. fragrans is rich in bioactive compounds and put forward several newly detected compounds for further investigation. Full article

(This article belongs to the Section Natural Products)

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20 pages, 3319 KiB

Open AccessArticle

Rational Design, Synthesis, and Biological Evaluation of Novel Thiazole/Thiazolidinones Multitarget Anti-Human Immunodeficiency Virus Molecules

by Christophe Tratrat, Anthi Petrou, Maria Fesatidou, Micheline Haroun, Mohamad Chohan and Athina Geronikaki

Pharmaceuticals 2025, 18(3), 298; https://doi.org/10.3390/ph18030298 - 21 Feb 2025

Abstract

Background: HIV-1 RT inhibitors were the first drugs approved to treat AIDS and remain key components of highly active antiretroviral therapy (HAART). While HAART effectively suppresses viral replication and slows disease progression, it has limitations, including long-term side effects and the emergence of [...] Read more.

Background: HIV-1 RT inhibitors were the first drugs approved to treat AIDS and remain key components of highly active antiretroviral therapy (HAART). While HAART effectively suppresses viral replication and slows disease progression, it has limitations, including long-term side effects and the emergence of drug-resistant strains, highlighting the need for new treatments. Objectives: Based on our previous experience, and insights from existing inhibitors of HIV-1 RT and RNase H, we aim to design and synthesize safer, multifunctional molecules. Methods: Using molecular docking studies, these compounds will incorporate pharmacophores targeting multiple stages of the HIV life cycle to enhance efficacy, reduce resistance, and improve pharmacokinetics. The compounds were synthesized via a one-pot three component reaction. The synthesized compounds were identified using spectroscopy and tested in vitro for activity against key HIV targets, including RNA-dependent DNA polymerase (RDDP) and RNAse H. Results: Among the synthesized compounds, several demonstrated strong inhibitory activity, with compound 11 showing IC₅₀ values comparable to the reference drug Nevirapine, and compound 4 exhibiting dual inhibition of both RT and RNase H activities. Conclusions: These findings emphasize the importance of a multidisciplinary approach, combining computational modeling with experimental validation, to identify promising leads for therapeutic development. Full article

(This article belongs to the Special Issue Pyrazole and Thiazole Derivatives in Medicinal Chemistry)

46 pages, 11937 KiB

Open AccessReview

Prodrug Approach as a Strategy to Enhance Drug Permeability

by Mateus Mello de Souza, Ana Luísa Rodriguez Gini, Jhonnathan Alves Moura, Cauê Benito Scarim, Chung Man Chin and Jean Leandro dos Santos

Pharmaceuticals 2025, 18(3), 297; https://doi.org/10.3390/ph18030297 - 21 Feb 2025

Abstract

Absorption and permeability are critical physicochemical parameters that must be balanced to achieve optimal drug uptake. These key factors are closely linked to the maximum absorbable dose required to provide appropriate plasma levels of drugs. Among the various strategies employed to enhance drug [...] Read more.

Absorption and permeability are critical physicochemical parameters that must be balanced to achieve optimal drug uptake. These key factors are closely linked to the maximum absorbable dose required to provide appropriate plasma levels of drugs. Among the various strategies employed to enhance drug solubility and permeability, prodrug design stands out as a highly effective and versatile approach for improving physicochemical properties and enabling the optimization of biopharmaceutical and pharmacokinetic parameters while mitigating adverse effects. Prodrugs are compounds with reduced or no activity that, through bio-reversible chemical or enzymatic processes, release an active parental drug. The application of this technology has led to significant advancements in drug optimization during the design phase, and it offers broad potential for further development. Notably, approximately 13% of the drugs approved by the U.S. Food and Drug Administration (FDA) between 2012 and 2022 were prodrugs. In this review article, we will explore the application of prodrug strategies to enhance permeability, describing examples of market drugs. We also describe the use of the prodrug approach to optimize PROteolysis TArgeting Chimeras (PROTACs) permeability by using conjugation technologies. We will highlight some new technologies in prodrugs to enrich permeability properties, contributing to developing new effective and safe prodrugs. Full article

(This article belongs to the Section Medicinal Chemistry)

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20 pages, 8476 KiB

Open AccessArticle

Harnessing Manuka Honey: A Natural Remedy for Accelerated Burn Wound Healing in a Porcine Model

by Boris Privrodski, Mladen Jovanović, Nikola Delić, Radomir Ratajac, Vladimir Privrodski, Aleksandar Stanojković, Bernadeta Gavlik and Ivan Čapo

Pharmaceuticals 2025, 18(3), 296; https://doi.org/10.3390/ph18030296 - 21 Feb 2025

Abstract

Backgrounds: Burn injuries present significant medical challenges due to their complexity in healing and potential for severe scarring. This study evaluates the efficacy of Manuka honey in accelerating burn wound healing compared to conventional antibiotic ointments. Methods: Using a porcine model resembling human [...] Read more.

Backgrounds: Burn injuries present significant medical challenges due to their complexity in healing and potential for severe scarring. This study evaluates the efficacy of Manuka honey in accelerating burn wound healing compared to conventional antibiotic ointments. Methods: Using a porcine model resembling human skin, nine Landrace breed female pigs with standardized deep dermal burns were treated with either Manuka honey in alginate or a combination of antibiotic ointments. Wound healing was assessed through macroscopic evaluation, a histopathological analysis, and immunohistochemical staining over a 60-day period. Results: Our findings indicate that the Manuka honey treatment was associated with significantly increased collagen density in the treated wounds compared to the control group (p < 0.05). The immunohistochemical analysis revealed lower macrophage activity (Iba1 staining) and a reduction in Ki67 expression on days 10 and 17 in the Manuka honey group, suggesting a more rapid transition toward tissue remodeling. The quantitative analysis showed a trend toward delayed epithelialization and increased inflammation in the control group, while wounds treated with Manuka honey exhibited faster reepithelialization and improved epidermal regeneration. However, additional studies are required to further assess collagen fiber organization and overall dermal architecture. Conclusions: These findings support the potential of Manuka honey as a beneficial treatment for burn wound healing, with evidence of enhanced reepithelialization and collagen deposition. Further research, including clinical trials, is necessary to fully elucidate its role in clinical practice and optimize treatment protocols. Full article

(This article belongs to the Special Issue Applications of Beehive Products for Wound Repair and Skin Care)

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14 pages, 3879 KiB

Open AccessArticle

PET Imaging Expedites Detection of Aberration in the Humanization of an Annexin A1 Targeting Antibody

by Hailey A. Houson, Brian D. Wright, Solana R. Fernandez, Tim Buss, Sharon L. White, Brittany Cederstrom, James M. Omweri, Jonathan E. McConathy, Jan E. Schnitzer and Suzanne E. Lapi

Pharmaceuticals 2025, 18(3), 295; https://doi.org/10.3390/ph18030295 - 21 Feb 2025

Abstract

Objectives: Annexin-A1 is a 37 kDa phospholipid-binding protein which is concentrated in a truncated 34 kDa form (AnnA1) in caveolae on the tumor vascular endothelial cell surface with expression in many tumor types. PRISM developed the monoclonal mouse antibody mAnnA1 against AnnA1 [...] Read more.

Objectives: Annexin-A1 is a 37 kDa phospholipid-binding protein which is concentrated in a truncated 34 kDa form (AnnA1) in caveolae on the tumor vascular endothelial cell surface with expression in many tumor types. PRISM developed the monoclonal mouse antibody mAnnA1 against AnnA1 for evaluation of AnnA1 as a potential target for imaging and therapy in oncology. mAnnA1 was humanized to make hAnnA1 for translation to clinical studies. Both PRISM-produced mAnnA1 and cGMP contractor-produced hAnnA1 were investigated using noninvasive PET/CT imaging, and dosimetry was evaluated to enable clinical translation of this strategy and to investigate in vivo behavior of hAnnA1. Methods: Antibodies mAnnA1 and hAnnA1 (PRISM “hAnnA1-P” or contractor generated “hAnnA1-C”) were conjugated with the chelator deferoxamine and evaluated for immunoreactivity with ELISA. Conjugated antibodies were radiolabeled with zirconium-89. Naïve mice, rats, and non-human primates (NHP) were injected with [⁸⁹Zr]mAnnA1 or [⁸⁹Zr]hAnnA1 and imaged with PET/CT up to 10 days post injection. After imaging, mice and rats were euthanized and organs were collected, weighed, and radioactivity was quantified using a gamma counter. Dosimetry in mice and NHPs were calculated using OLINDA. Results: [⁸⁹Zr]mAnnA1 showed similar biodistribution to other antibodies with slow clearance through the liver. Transition to [⁸⁹Zr]hAnnA1-C during the dosimetry studies revealed substantial uptake in the spleen (130 ± 48% ID/g at day 5 post injection in female BALB/c), which was not observed with [⁸⁹Zr]mAnnA1 (5.6 ± 1.7% ID/g at day 7 PI). Further studies in multiple strains of mice showed variable elevated splenic uptake of [⁸⁹Zr]hAnnA1-C across mouse strains, with the highest uptake observed in female BALB/c mice (118.4 ± 23.1% ID/g) and the lowest uptake observed in male CD1 mice (34.7 ± 10.2% ID/g). Additionally, splenic uptake of hAnnA1-C was observed in Fischer rats (2.8 ± 0.6% ID/organ) and NHPs (1.6 ± 0.6% ID/organ), although at lower levels than what was observed in BALB/c mice (8.8 ± 1.8% ID/organ). Dosimetry results showed similar values between estimates based on mouse and NHP data, with the largest difference seen in the spleen (5.2 vs. 2.6 mSv/MBq in females respectively). Sequencing of hAnnA1-C revealed a frameshift mutation in the antibody sequence introduced during cGMP manufacture. Restoration of the antibody sequence by PRISM returned the antibody distribution into alignment with mAnnA1. Conclusions: An aberration introduced during cGMP production of hAnnA1-C resulted in increased splenic uptake and alteration of the biodistribution in mice. PET imaging enabled quantitative detection of the immunogenic behavior of hAnnA1, which led to detection of the sequence error. Restoration of the sequence resulted in an antibody which was non-immunogenic to mice. Full article

(This article belongs to the Special Issue Challenges in the Development of PET and SPECT Radiotracers: Innovative Ideas with Negative Outcomes)

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16 pages, 7196 KiB

Open AccessArticle

Exploring the Potential Mechanism of Liupao Tea Using UPLC-Q-TOF/MS and Network Pharmacology

by Fang Jia, Qi Yang, Lihao Yao, Yunfei Liu, Jiagang Deng, Jing Leng, Lili Fan and Erwei Hao

Pharmaceuticals 2025, 18(3), 294; https://doi.org/10.3390/ph18030294 - 21 Feb 2025

Abstract

Background: Gastrointestinal motility disorder (GMD) is a common condition characterized by dysfunction or degeneration of the myenteric plexus in specific segments of the gastrointestinal tract. Liupao tea (LPT) is a post-fermented tea that is rich in various secondary metabolites and has demonstrated a [...] Read more.

Background: Gastrointestinal motility disorder (GMD) is a common condition characterized by dysfunction or degeneration of the myenteric plexus in specific segments of the gastrointestinal tract. Liupao tea (LPT) is a post-fermented tea that is rich in various secondary metabolites and has demonstrated a range of pharmacological effects, including lipid-lowering properties, antioxidant activity, and modulation of the gut microbiota. However, the underlying mechanisms by which LPT improves GMD remain poorly understood. Methods: Blood was collected after gavage of LPT extract in SD rats. The active components in the aqueous extract of LPT and its serum were analyzed using ultra-high-performance liquid chromatography quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The targets of LPT in the treatment of GMD were predicted by network pharmacology and molecular docking. Results: 65 compounds were identified in the water extract of LPT, including flavonoids, phenolic acids, alkaloids, and amino acids. In rats treated with LPT, 14 prototype compounds and 6 metabolites were detected in serum. Network pharmacology and molecular docking analyses revealed 298 common targets between LPT and GMD, including IL-6, AKT1, and TP53. Functional enrichment analysis suggested that LPT may improve GMD through the regulation of immune, inflammatory, and cytokine signaling pathways. Molecular docking further indicated that the primary bioactive components of LPT exhibit a strong affinity for IL-6, AKT1, and TP53. Conclusions: These findings provide new insights into the bioactive components, molecular targets, and mechanisms of LPT, suggesting its potential as a therapeutic strategy for gastrointestinal motility disorders. Full article

(This article belongs to the Section Natural Products)

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25 pages, 1838 KiB

Open AccessReview

Preclinical Retinal Disease Models: Applications in Drug Development and Translational Research

by Sudha Priya Soundara Pandi, Hanagh Winter, Madeleine R. Smith, Kevin Harkin and James Bojdo

Pharmaceuticals 2025, 18(3), 293; https://doi.org/10.3390/ph18030293 - 21 Feb 2025

Abstract

Retinal models play a pivotal role in translational drug development, bridging preclinical research and therapeutic applications for both ocular and systemic diseases. This review highlights the retina as an ideal organ for studying advanced therapies, thanks to its immune privilege, vascular and neuronal [...] Read more.

Retinal models play a pivotal role in translational drug development, bridging preclinical research and therapeutic applications for both ocular and systemic diseases. This review highlights the retina as an ideal organ for studying advanced therapies, thanks to its immune privilege, vascular and neuronal networks, accessibility, and advanced imaging capabilities. Preclinical retinal disease models offer unparalleled insights into inflammation, angiogenesis, fibrosis, and hypoxia, utilizing clinically translatable bioimaging tools like fundoscopy, optical coherence tomography, confocal scanning laser ophthalmoscopy, fluorescein angiography, optokinetic tracking, and electroretinography. These models have driven innovations in anti-inflammatory, anti-angiogenic, and neuroprotective strategies, with broader implications for systemic diseases such as rheumatoid arthritis, Alzheimer’s, and fibrosis-related conditions. By emphasizing the integration of the 3Rs principles and novel imaging modalities, this review highlights how retinal research not only enhances therapeutic precision but also minimizes ethical concerns, paving the way for more predictive and human-relevant approaches in drug development. Full article

(This article belongs to the Special Issue Novel Treatments and Technologies for Retinal Diseases)

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22 pages, 5699 KiB

Open AccessArticle

Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias

by Anqi Chen, Chenxi Su, Zisu Zhang and Haitao Zhang

Pharmaceuticals 2025, 18(3), 292; https://doi.org/10.3390/ph18030292 - 21 Feb 2025

Abstract

Background: The four subtypes of G protein-coupled receptors (GPCRs) regulated by histamine play critical roles in various physiological and pathological processes, such as allergy, gastric acid secretion, cognitive and sleep disorders, and inflammation. Previous experimental structures of histamine receptors (HRs) with agonists and [...] Read more.

Background: The four subtypes of G protein-coupled receptors (GPCRs) regulated by histamine play critical roles in various physiological and pathological processes, such as allergy, gastric acid secretion, cognitive and sleep disorders, and inflammation. Previous experimental structures of histamine receptors (HRs) with agonists and antagonists exhibited multiple conformations for the ligands and G protein binding. However, the structural basis for HR regulation and signaling remains elusive. Methods: We determined the cryo-electron microscopy (cryo-EM) structure of the H4R-histamine-Gi complex at 2.9 Å resolution, and predicted the models for all four HRs in the ligand-free apo and G protein subtype binding states using AlphaFold3 (AF3). Results: By comparing our H4R structure with the experimental HR structures and the computational AF3 models, we elucidated the distinct histamine binding modes and G protein interfaces, and proposed the essential roles of Y^6.51 and Q^7.42 in receptor activation and the intracellular loop 2 (ICL2) in G protein bias. Conclusions: Our findings deciphered the molecular mechanisms underlying the regulation of different HRs, from the extracellular ligand-binding pockets and transmembrane motifs to the intracellular G protein coupling interfaces. These insights are expected to facilitate selective drug discovery targeting HRs for diverse therapeutic purposes. Full article

(This article belongs to the Special Issue Histamine Receptor Ligands in Medicinal Chemistry)

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16 pages, 1367 KiB

Open AccessReview

2024 FDA TIDES (Peptides and Oligonucleotides) Harvest

by Othman Al Musaimi, Danah AlShaer, Beatriz G. de la Torre and Fernando Albericio

Pharmaceuticals 2025, 18(3), 291; https://doi.org/10.3390/ph18030291 - 20 Feb 2025

Abstract

In 2024, the FDA approved fifty novel drugs, including four peptides and oligonucleotides (TIDEs) (two pepTIDEs and two oligonucleoTIDEs), highlighting their increasing importance as effective alternatives to traditional drug classes. TIDEs provide essential therapies for complex diseases, such as genetic disorders, rather than [...] Read more.

In 2024, the FDA approved fifty novel drugs, including four peptides and oligonucleotides (TIDEs) (two pepTIDEs and two oligonucleoTIDEs), highlighting their increasing importance as effective alternatives to traditional drug classes. TIDEs provide essential therapies for complex diseases, such as genetic disorders, rather than merely addressing symptoms. In addition to oligonucleotide therapeutics for various genetic conditions, peptides became the first approved treatment for Rett Syndrome in 2023 and were also used to treat Niemann–Pick disease type C (NPC) in 2024. Interestingly, among the strategies employed in recent approvals to enhance stability and/or delivery, the prodrug approach, exemplified by palopegteriparatide and pegulicianine, is emerging as a more targeted and precise therapeutic strategy. Additionally, the Enhanced Stabilization Chemistry (ESC)-GalNAc platform has been expanded for hepatic delivery of a new oligonucleotide drug, olezarsen. Furthermore, novel modifications to the ribose moiety in oligonucleotides, such as the 3′-amino substitution in imetelstat, enhance their stability. This review examines the TIDES approved in 2024 based on their chemical structure, medical targets, modes of action, administration routes, and common adverse effects. In addition, it highlights how the prodrug strategy has improved targeting efficiency and extended the half-lives of the active drugs. Full article

(This article belongs to the Section Biopharmaceuticals)

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39 pages, 6209 KiB

Open AccessArticle

Optimized Hesperidin-Loaded Lipid Nanoparticles with Tea Tree Oil for Enhanced Wound Healing: Formulation, Characterization, and Evaluation

by Borros Arneth, Rehab Abdelmonem, Mohamed A. El-Nabarawi, Mahmoud Hassan Teaima, Kareem Omar Rashwan, Mohamed A. Soliman and Inas Essam Ibrahim Al-Samadi

Pharmaceuticals 2025, 18(3), 290; https://doi.org/10.3390/ph18030290 - 20 Feb 2025

Abstract

Objectives: This study aimed to develop hesperidin solid lipid nanoparticles (HESP-SLNs) to enhance their stability, solubility, and sustained release for wound healing; further enhancement was achieved through prepared nanostructured lipid carriers (HESP-NLCs) using Tea Tree Oil (TTO) to explore their synergistic efficacy. Methods [...] Read more.

Objectives: This study aimed to develop hesperidin solid lipid nanoparticles (HESP-SLNs) to enhance their stability, solubility, and sustained release for wound healing; further enhancement was achieved through prepared nanostructured lipid carriers (HESP-NLCs) using Tea Tree Oil (TTO) to explore their synergistic efficacy. Methods: A factorial design of 2⁴ trials was established to evaluate the influence of lipid type (X1), lipid conc (%) (X2), surfactant type (X3), and sonication amplitude (%) (X4) of prepared HESP-SLNs on the particle size (nm) (Y1), polydispersibility index (Y2), zeta potential (Y3), and encapsulation efficiency (%) (Y4). The optimized HESP-SLNs formula was selected utilizing Design Expert^® software version 13, which was additionally enhanced by preparing TTO-loaded HESP-NLCs. In vitro release, Raman spectroscopy, and transmission electron microscopy were carried out for both lipid nanoparticles. Cytotoxicity, in vivo wound-healing assessments, and skin irritancy tests were performed to evaluate the performance of TTO-incorporated HESP-NLCs compared to HESP-SLNs. Results: The optimized formula demonstrated PS (280 ± 1.35 nm), ZP (−39.4 ± 0.92 mV), PDI (0.239 ± 0.012), and EE% (88.2 ± 2.09%). NLCs enhanced Q6% release, (95.14%) vs. (79.69%), for SLNs and showed superior antimicrobial efficacy. Both lipid nanoparticles exhibited spherical morphology and compatibility between HESP and excipients. NLCs achieved the highest wound closure percentage, supported by histological analysis and inflammatory biomarker outcomes. Cytotoxicity evaluation showed 87% cell viability compared to untreated HSF cells, and the skin irritancy test confirmed the safety of NLCs. Conclusions: TTO-loaded HESP-NLCs are promising candidates exhibiting superior wound-healing capabilities, making them a potential therapeutic option for cutaneous wound management. Full article

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

13 pages, 242 KiB

Open AccessReview

Protegrin-1 and Analogues Against Acinetobacter baumannii: A Narrative Review

by Giulio Rizzetto, Edoardo De Simoni, Elisa Molinelli, Cecilia Busignani, Corrado Tagliati, Daisy Gambini, Annamaria Offidani and Oriana Simonetti

Pharmaceuticals 2025, 18(3), 289; https://doi.org/10.3390/ph18030289 - 20 Feb 2025

Abstract

A. baumannii is recognised as an important etiologic agent for hospital infections and increases the risk of postoperative complications, worsening mortality and prolonging hospitalisation. Protegrin-1 (PG-1) is one of the most promising antimicrobial peptides (AMPs) in the literature, since its antimicrobial action covers [...] Read more.

A. baumannii is recognised as an important etiologic agent for hospital infections and increases the risk of postoperative complications, worsening mortality and prolonging hospitalisation. Protegrin-1 (PG-1) is one of the most promising antimicrobial peptides (AMPs) in the literature, since its antimicrobial action covers a wide range of Gram-positive and Gram-negative bacteria, including A. baumannii. PG-1 represents a valid new therapeutic option for the treatment of A. baumannii multi-drug resistant infections, showing synergic activity with traditional antibiotics, such as colistin. However, its clinical use in humans still requires studies, especially considering the haemolytic risk. For this reason, the use of PG-1 analogues, such as PLP-3, HV2, CDP-1, and IB367, seems to be the most promising way for the clinical use of this class of AMPs. Full article

(This article belongs to the Section Medicinal Chemistry)

31 pages, 4769 KiB

Open AccessArticle

Formulation Development of Natural Polymeric Nanoparticles, In Vitro Antiaging Evaluation, and Metabolite Profiling of Toona sinensis Leaf Extracts

by Uce Lestari, Muhaimin Muhaimin, Anis Yohana Chaerunisaa and Wawan Sujarwo

Pharmaceuticals 2025, 18(3), 288; https://doi.org/10.3390/ph18030288 - 20 Feb 2025

Abstract

Background/Objectives: Natural polymer nanoparticles have potential as delivery systems, can enhance pharmacological activity, and can improve stability in the cosmetic field. In this research, we implemented a development approach for chitosan–alginate and chitosan–pectin nanoparticles. This study aimed to investigate effect of formulation, process [...] Read more.

Background/Objectives: Natural polymer nanoparticles have potential as delivery systems, can enhance pharmacological activity, and can improve stability in the cosmetic field. In this research, we implemented a development approach for chitosan–alginate and chitosan–pectin nanoparticles. This study aimed to investigate effect of formulation, process variables, in vitro antiaging evaluation, and metabolite profiling of Toona sinensis leaf extracts. Methods: Polymeric nanoparticles have been prepared using the ionic gelation method (Temperature = 40 °C, time = 1 h and speed = 1000 rpm), in vitro antiaging evaluation using the Neutrophil Elastase Inhibitor Screening Kit method, and analysis of metabolite profiling with UHPLC–HRMS. Results: Research results found that the SLE and EAFSL nanoparticles that have good and stable characteristics before and after storage in a climatic chamber after 3 months are FIIA-NPSLE (0.75% chitosan and 1.25% Alginate), FIP-NPSLE (1% chitosan and 0.5% Pectin), FIIA-NPEAFSL (0.75% chitosan and 1.25% Alginate), and FIIIP-NPEAFSL (0.125% chitosan and 0.375% Alginate). Chitosan–alginate polymers, such as FIIA-NPEAFSL, have higher inhibition of the elastase enzyme than FIIA-NPSLE, with a % inhibition (IC₅₀) of FIIA-NPEAFSL being 87.30%, while the IC₅₀ of FIIA-NPSLE is 39.40%. Meanwhile, using chitosan–pectin polymers, such as FIP-NPSLE, results in lower inhibition of the elastase enzyme compared to the chitosan–alginate polymer, with an IC₅₀ of 27.28% while IC₅₀ FIIIP-NPEAFSL is 39.53%. SLE and EAFSL nanoparticles with chitosan–alginate and chitosan–pectin polymers resulted in a significant PDI during storage from 1.3 to 1.9, and zeta potential values were very low, ranging from −11 mV to −27 mV. Metabolite profiling using UHPLC–HRMS on T. sinensis leaf extracts revealed that the main compounds contained were glycitein, quercetin, quercetin-3β-D-glucoside, kaempferol, and ellagic acid, which has potential as an antiaging agent. Conclusions: It can be concluded that using chitosan, alginate, and pectin in the process of encapsulating extracts into nanoparticles with the same process variables affect evaluation of antiaging activity in elastase enzymes. Further research will develop these nanoparticles into nanohydrogels with antiaging activity. Full article

(This article belongs to the Section Pharmaceutical Technology)

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18 pages, 10138 KiB

Open AccessArticle

Micheliolide Alleviates Hepatic Fibrosis by Inhibiting Autophagy in Hepatic Stellate Cells via the TrxR1/2-Mediated ROS/MEK/ERK Pathway

by Yi Liu, Ling Yao, Yuanyuan Liu, Yunheng Yang, Ailing Liang, Honglin He, Yao Lei, Wenfu Cao and Zhiwei Chen

Pharmaceuticals 2025, 18(3), 287; https://doi.org/10.3390/ph18030287 - 20 Feb 2025

Abstract

Background: Hepatic fibrosis is a major global health issue without an optimal drug treatment, highlighting the urgent need to find effective therapies. This study aimed to clarify the role and mechanism of micheliolide in treating hepatic fibrosis. Methods: The efficacy of [...] Read more.

Background: Hepatic fibrosis is a major global health issue without an optimal drug treatment, highlighting the urgent need to find effective therapies. This study aimed to clarify the role and mechanism of micheliolide in treating hepatic fibrosis. Methods: The efficacy of MCL was evaluated in a mouse model of CCl₄-induced hepatic fibrosis. LX-2 cells were subjected to MCL treatment, and subsequent changes in fibrosis markers, autophagy, and the MEK/ERK pathway were analyzed using transcriptomics and Western blotting. The interaction between MCL and TrxR1 or TrxR2 were validated using cellular thermal shift assays (CETSA) and drug affinity responsive target stability (DARTS) assays. Results: Our findings indicated that MCL significantly alleviated CCl₄-induced hepatic fibrosis, improved liver function, and downregulated the expression of fibrosis markers. Additionally, MCL significantly inhibited LX-2 cell activation by suppressing cell proliferation, extracellular matrix (ECM) production, and autophagy, while activating the MEK/ERK pathway. Moreover, MCL elevated intracellular and mitochondrial reactive oxygen species (ROS) levels, reduced mitochondrial membrane potential, and altered mitochondrial morphology. The ROS scavenger N-acetylcysteine (NAC) attenuated MCL-induced MEK/ERK pathway activation and increased collagen type I alpha 1 (COL1A1) and fibronectin (FN) expression. Further analysis confirmed that MCL directly interacts with TrxR1 and TrxR2, leading to the inhibition of their enzymatic activities and the induction of ROS generation. Ultimately, MCL attenuated the fibrotic process and autophagic flux in LX-2 cells. Conclusions: The findings of our study confirmed that MCL has the potential to alleviate hepatic fibrosis, thereby introducing a novel candidate drug and therapeutic strategy for management of this condition. Full article

(This article belongs to the Section Pharmacology)

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45 pages, 9208 KiB

Open AccessReview

Therapeutic Potential of Plant- and Marine-Derived Bioactive Compounds in Prostate Cancer: Mechanistic Insights and Translational Applications

by Arif Jamal Siddiqui, Mohd Adnan, Juhi Saxena, Mohammad Jahoor Alam, Abdelmushin Abdelgadir, Riadh Badraoui and Ritu Singh

Pharmaceuticals 2025, 18(3), 286; https://doi.org/10.3390/ph18030286 - 20 Feb 2025

Abstract

It is widely recognized that prostate cancer is a multifaceted illness that is the second most common cause of cancer-related fatalities among males. Natural sources from both plants and marine organisms have long been used in treating various diseases and in the discovery [...] Read more.

It is widely recognized that prostate cancer is a multifaceted illness that is the second most common cause of cancer-related fatalities among males. Natural sources from both plants and marine organisms have long been used in treating various diseases and in the discovery of new pharmaceutical compounds. Medicinal plants, in particular, provide bioactive substances like alkaloids, phenolic compounds, terpenes, and steroids. In addition, marine natural products play a crucial role in the search for novel cancer treatments. A substantial number of anticancer drugs have been derived from natural sources, including plants, marine organisms, and microorganisms. In fact, over the past 60 years, 80% of new chemical entities have originated from natural sources, which are generally considered safer than synthetic compounds. This review seeks to emphasize the role of phytochemical compounds derived from both plant and marine sources in prostate cancer, highlighting their potential therapeutic impact. It is also intended to support global researchers working on the identification of natural-based treatments for prostate cancer. Full article

(This article belongs to the Special Issue Exploring Natural Products with Antioxidant and Anticancer Properties)

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14 pages, 1361 KiB

Open AccessArticle

Implementation of the Anchor-Based Indirect Comparison Method for Equivalence Margin Derivation in Biosimilar Development

by Claudia Hemmelmann, Rachid El Galta, Jessie Wang, Susanne Schmitt and Ramin Arani

Pharmaceuticals 2025, 18(3), 285; https://doi.org/10.3390/ph18030285 - 20 Feb 2025

Abstract

Background/Objectives: To derive the equivalence margin (EQM), typically, a “classical” meta-analysis on direct within-trial estimation of the effect size of the reference drug compared to the placebo or standard of care is performed: a certain factor of the 95% confidence interval for [...] Read more.

Background/Objectives: To derive the equivalence margin (EQM), typically, a “classical” meta-analysis on direct within-trial estimation of the effect size of the reference drug compared to the placebo or standard of care is performed: a certain factor of the 95% confidence interval for the pooled treatment effect compared to placebo is used. However, treatment regimens in many indications are becoming more complex (e.g., combination treatments), and for most of these clinical study data, direct comparisons are not available. On the other hand, data for the comparison of the common treatment to the reference treatment in one study and to the placebo in another study are available in some situations. Methods: In such situations, an anchor-based indirect comparison can be applied to estimate the treatment effect of Reference vs. Placebo. This treatment effect (Reference vs. Placebo) can be estimated by calculating the difference of the two treatment effects and the variance as the sum of both variances. The 95% confidence interval of this estimated treatment effect can then be used to derive the EQM. To alleviate any concerns about the underlying assumptions of transitivity and consistency, multiple sensitivity analyses can be performed. Results: We present a case study for deriving the EQM using the anchor-based indirect comparison along with sensitivity analyses (i.e., direct comparison against similar reference drug, the impact of variation of treatment effect on Comparator, and effect size Reference vs. Placebo, including trial data with slightly different population characteristics) for a planned efficacy trial in the biosimilar setting. Conclusions: An anchor-based indirect comparison for EQM derivation is an approach health authorities can agree to if sufficiently supported through other means, e.g., relevant sensitivity analyses. Full article

(This article belongs to the Topic Biosimilars and Interchangeability)

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20 pages, 3725 KiB

Open AccessReview

Rethinking Clinical Trials in Age-Related Macular Degeneration: How AI-Based OCT Analysis Can Support Successful Outcomes

by Marie Louise Enzendorfer, Merle Tratnig-Frankl, Anna Eidenberger, Johannes Schrittwieser, Lukas Kuchernig and Ursula Schmidt-Erfurth

Pharmaceuticals 2025, 18(3), 284; https://doi.org/10.3390/ph18030284 - 20 Feb 2025

Abstract

Age-related macular degeneration (AMD) is a leading cause of blindness in the developed world. Due to an aging population, its prevalence is expected to increase, making novel and optimized therapy options imperative. However, both late-stage forms of the disease, neovascular AMD (nAMD) and [...] Read more.

Age-related macular degeneration (AMD) is a leading cause of blindness in the developed world. Due to an aging population, its prevalence is expected to increase, making novel and optimized therapy options imperative. However, both late-stage forms of the disease, neovascular AMD (nAMD) and geographic atrophy (GA), exhibit considerable variability in disease progression and treatment response, complicating the evaluation of therapeutic efficacy and making it difficult to design clinical trials that are both inclusive and statistically robust. Traditional trial designs frequently rely on generalized endpoints that may not fully capture the nuanced benefits of treatment, particularly in diseases like GA, where functional improvements can be gradual or subtle. Artificial intelligence (AI) has the potential to address these issues by identifying novel, condition-specific biomarkers or endpoints, enabling precise patient stratification and improving recruitment strategies. By providing an overview of the advances and application of AI-based optical coherence tomography analysis in the context of AMD clinical trials, this review highlights the transformative potential of AI in optimizing clinical trial outcomes for patients with nAMD or GA secondary to AMD. Full article

(This article belongs to the Special Issue Novel Treatments and Technologies for Retinal Diseases)

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22 pages, 3558 KiB

Open AccessArticle

Dengue Virus Inhibitors as Potential Broad-Spectrum Flavivirus Inhibitors

by Larisa Ivanova, Krystyna Naumenko, Margus Varjak, Sandra Koit, Yehudit Morozovsky, Andres Merits, Mati Karelson and Eva Zusinaite

Pharmaceuticals 2025, 18(3), 283; https://doi.org/10.3390/ph18030283 - 20 Feb 2025

Abstract

Background. Flaviviruses spread from endemic to non-endemic areas, causing illness in millions of people worldwide. The lack of effective therapies and the rapid expansion of flaviviral infections worldwide emphasize the importance of finding effective antivirals to treat such diseases. Objectives. To find out [...] Read more.

Background. Flaviviruses spread from endemic to non-endemic areas, causing illness in millions of people worldwide. The lack of effective therapies and the rapid expansion of flaviviral infections worldwide emphasize the importance of finding effective antivirals to treat such diseases. Objectives. To find out the potential broad-spectrum flavivirus inhibitors among previously reported inhibitors of DENV2/DENV4. Methods. The cytotoxicity of compounds was tested using WST-1 assay. The compounds were tested for their ability to inhibit the infection of DENV2, ZIKV, KUNV, and TBEV, and the most active compounds were also analyzed using the replicon-based assay. Interactions of one of the identified inhibitors with possible viral targets were studied using molecular dynamics simulations. Results. Two out of eight previously reported DENV2/DENV4 inhibitors demonstrated the ability to inhibit all studied viruses at low micromolar concentrations. Compound C6 demonstrated the ability to inhibit both DENV2 and TBEV. Compounds C1 (lycorine), C3 (mycophenolic acid), and C7 (vidarabine) were demonstrated as inhibitors of TBEV infection for the first time. Conclusions. Several compounds, previously described as inhibitors of DENV, are also able to inhibit other flaviviruses. This work is the first report on the anti-TBEV activity of lycorine (C1) and mycophenolic acid (C3), as well as vidarabine (C7). In addition, this is the first experimental confirmation of the antiviral activity of compound C5 and the lack of detectable antiviral activity of compound C8, demonstrating the necessity of experimental verification of the computational predictions. Full article

(This article belongs to the Special Issue Antiviral Agents, 2024)

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29 pages, 6612 KiB

Open AccessReview

Artificial Intelligence Models and Tools for the Assessment of Drug–Herb Interactions

by Marios Spanakis, Eleftheria Tzamali, Georgios Tzedakis, Chryssalenia Koumpouzi, Matthew Pediaditis, Aristides Tsatsakis and Vangelis Sakkalis

Pharmaceuticals 2025, 18(3), 282; https://doi.org/10.3390/ph18030282 - 20 Feb 2025

Abstract

Artificial intelligence (AI) has emerged as a powerful tool in medical sciences that is revolutionizing various fields of drug research. AI algorithms can analyze large-scale biological data and identify molecular targets and pathways advancing pharmacological knowledge. An especially promising area is the assessment [...] Read more.

Artificial intelligence (AI) has emerged as a powerful tool in medical sciences that is revolutionizing various fields of drug research. AI algorithms can analyze large-scale biological data and identify molecular targets and pathways advancing pharmacological knowledge. An especially promising area is the assessment of drug interactions. The AI analysis of large datasets, such as drugs’ chemical structure, pharmacological properties, molecular pathways, and known interaction patterns, can provide mechanistic insights and identify potential associations by integrating all this complex information and returning potential risks associated with these interactions. In this context, an area where AI may prove valuable is in the assessment of the underlying mechanisms of drug interactions with natural products (i.e., herbs) that are used as dietary supplements. These products pose a challenging problem since they are complex mixtures of constituents with diverse and limited information regarding their pharmacological properties, especially their pharmacokinetic data. As the use of herbal products and supplements continues to grow, it becomes increasingly important to understand the potential interactions between them and conventional drugs and the associated adverse drug reactions. This review will discuss AI approaches and how they can be exploited in providing valuable mechanistic insights regarding the prediction of interactions between drugs and herbs, and their potential exploitation in experimental validation or clinical utilization. Full article

(This article belongs to the Special Issue Therapeutic Drug Monitoring and Adverse Drug Reactions: 2nd Edition)

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