Pharmaceuticals

37 pages, 10612 KB

Open AccessReview

Chiral-Modified Nucleoside Analogues: From Bioactivity to Therapeutic Applications

by Anna A. Kozlova, Valentina N. Borokh, Vladimir E. Oslovsky, Cyril S. Alexeev and Mikhail S. Drenichev

Pharmaceuticals 2026, 19(7), 1131; https://doi.org/10.3390/ph19071131 - 22 Jul 2026

Viewed by 619

Abstract

Nucleosides are extensively employed for the development of pharmaceuticals, chemotherapeutic agents, and bioregulators. Background/Objectives: The introduction of an additional chiral functionality into a carbohydrate or heterocyclic base fragment may increase the selectivity of interactions with nucleos(t)ide-metabolizing enzymes and receptors and, in some [...] Read more.

Nucleosides are extensively employed for the development of pharmaceuticals, chemotherapeutic agents, and bioregulators. Background/Objectives: The introduction of an additional chiral functionality into a carbohydrate or heterocyclic base fragment may increase the selectivity of interactions with nucleos(t)ide-metabolizing enzymes and receptors and, in some cases, lead to more specific physiological activities. Methods: An improvement of the selectivity of nucleoside-based drugs can be achieved either by the chemical modification of a carbohydrate or a base constituent or by a combination of these two approaches. Additionally, stereospecific enzymatic cleavage of nucleos(t)ide prodrugs containing biodegradable substituents can reduce cytotoxicity and enhance bioavailability. Results: A series of enantiomerically pure nucleosides modified at the ribose or heterocyclic base were obtained by chemical and enzymatic methods. Novel antiviral or anticancer active compounds, inhibiting viral or cellular enzymes or activating cellular nucleoside kinases have been found among chemically synthesized derivatives. Some exhibit strengthened “ligand–receptor” interaction, acting on receptors of the purinergic signaling system. During recent extensive structure–activity studies, several drugs and their prototypes have been proposed for the treatment of viral infections: the 2′C-fluoromethyl derivative of sofosbuvir (anti-SARS-CoV, preclinical), VV-261 (SFTSV, phase I clinical), balapiravir (dengue, phase I clinical), mericitabine (approved drug for HCV), and lumicitabine (approved drug for RSV and HMPV). Conclusions: Modern literature data within the scope of the present review suggest that direct modification of nucleosides with various chiral functionalities can be considered as an approach to increase their efficacy, specificity and selectivity. Full article

(This article belongs to the Special Issue Chirality in Drugs)

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17 pages, 11819 KB

Open AccessArticle

Juglone Protects Against CLP-Induced Sepsis by Regulating Apoptosis, Pyroptosis, and Oxidative Stress Mechanisms

by Ömer Faruk Başer and Mahmut Karapehlivan

Pharmaceuticals 2026, 19(7), 1130; https://doi.org/10.3390/ph19071130 - 22 Jul 2026

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Abstract

Background: Sepsis is a life-threatening systemic condition characterized by organ dysfunction resulting from a dysregulated host response to infection. This study aimed to investigate the protective role of juglone (5-hydroxy-1,4-naphthoquinone), a naturally occurring compound, on lung tissue in a cecal ligation and [...] Read more.

Background: Sepsis is a life-threatening systemic condition characterized by organ dysfunction resulting from a dysregulated host response to infection. This study aimed to investigate the protective role of juglone (5-hydroxy-1,4-naphthoquinone), a naturally occurring compound, on lung tissue in a cecal ligation and puncture (CLP)-induced sepsis model. Methods: Male Wistar-albino rats were used to establish the model, and juglone was administered intraperitoneally at doses of 1, 2, and 3 mg/kg. Lung and serum samples were collected for biochemical, molecular, and histological analyses through ELISA, RT-PCR, Western blot, and histopathological examinations. Results: In the sepsis group, the levels of proinflammatory cytokines (IL-1β, IL-6, IL-18) and pyroptosis-related markers (NLRP3, caspase-1, GSDMD) were significantly elevated, while juglone pretreatment markedly reduced these parameters in a dose-dependent manner. Moreover, juglone upregulated the Nrf2/HO-1 antioxidant pathway while downregulating Keap1 expression. RT-PCR analysis revealed that juglone suppressed the expression of pro-apoptotic genes (caspase-3, caspase-9, Bax) and enhanced anti-apoptotic Bcl-2 expression. Histopathological evaluation demonstrated that juglone alleviated inflammatory cell infiltration, septal thickening, and hemorrhage in lung tissue. Conclusions: These findings suggest that juglone is associated with protection against sepsis-induced lung injury and with changes in oxidative stress, inflammation, apoptosis, and pyroptosis pathways. Therefore, juglone may have protective potential against sepsis-induced pulmonary damage. Full article

(This article belongs to the Section Natural Products)

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17 pages, 1671 KB

Open AccessArticle

Phytochemical Composition and In Vitro Anti-Pigmentation Activity of Persicaria senticosa Flower Absolute: Potential Dual Inhibition of Melanogenesis and Melanosome Transport

by Kyung Jong Won, Hwan Myung Lee, Yoon Yi Kim, Ji Hye Bae, Ji Seong Yun and Do Yoon Kim

Pharmaceuticals 2026, 19(7), 1129; https://doi.org/10.3390/ph19071129 - 22 Jul 2026

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Abstract

Background/Objectives: Persicaria senticosa (Meisn.) H.Gross (PS) has anti-photoaging, anti-inflammatory, and antioxidant activities, but the anti-pigmentation potential of the PS flower absolute (PSFAb) remains largely unexplored. This study aimed to examine the chemical composition and anti-melanogenic and melanosome transport-inhibitory effects of PSFAb using [...] Read more.

Background/Objectives: Persicaria senticosa (Meisn.) H.Gross (PS) has anti-photoaging, anti-inflammatory, and antioxidant activities, but the anti-pigmentation potential of the PS flower absolute (PSFAb) remains largely unexplored. This study aimed to examine the chemical composition and anti-melanogenic and melanosome transport-inhibitory effects of PSFAb using B16BL6 murine melanoma cells. Methods: PSFAb was extracted with hexane and analyzed by gas chromatography–mass spectrometry (GC-MS). The biological activities in B16BL6 murine melanoma cells were evaluated using water-soluble tetrazolium salt (WST) assays, 5-bromo-2′-deoxyuridine (BrdU) incorporation, enzyme-linked immunosorbent assays, and immunoblotting methods. Results: GC-MS analysis identified eight constituents in PSFAb. Cell viability was not significantly altered in B16BL6 cells at concentrations ≤ 100 μg/mL, which were used for additional tests. PSFAb inhibited serum-induced cell proliferation and suppressed α-melanocyte-stimulating hormone (α-MSH)-induced melanin synthesis and tyrosinase activity in B16BL6 cells. PSFAb also downregulated the α-MSH-induced expression of key melanogenic regulators, including microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2. PSFAb decreased extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase phosphorylation but enhanced JNK phosphorylation in α-MSH-stimulated B16BL6 cells. Furthermore, PSFAb reduced the α-MSH-induced expression of melanosome transport-related proteins (melanophilin and Rab27a) in B16BL6 cells. Conclusions: Overall, these results suggest that PSFAb has the potential to exert anti-pigmentation effects by suppressing melanogenesis and downregulating melanosome transport-related proteins. Therefore, PSFAb may be a promising candidate for the development of natural agents targeting hyperpigmentation and skin pigmentation regulation. Full article

(This article belongs to the Section Natural Products)

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18 pages, 3618 KB

Open AccessArticle

Twenty-Year Trends in Antipsychotic Utilization in Serbia: A Nationwide Drug Utilization Study

by Zorana Pavlovic, Milena Stevanovic, Marija Milic, Jelena Filimonovic, Bojana Matejić, Mladen Bogdanovic, Ivana Vukajlovic, Aleksandar Krstić, Miodrag Milenović and Bojana Dunjić Kostić

Pharmaceuticals 2026, 19(7), 1128; https://doi.org/10.3390/ph19071128 - 21 Jul 2026

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Abstract

Background/Objectives: Antipsychotic drug utilization has changed substantially over recent decades, reflecting evolving prescribing practices, drug availability, and treatment guidelines. Methods: This study analyzed national antipsychotic consumption in Serbia from 2006 to 2024 using official data from the Medicines and Medical Devices Agency of [...] Read more.

Background/Objectives: Antipsychotic drug utilization has changed substantially over recent decades, reflecting evolving prescribing practices, drug availability, and treatment guidelines. Methods: This study analyzed national antipsychotic consumption in Serbia from 2006 to 2024 using official data from the Medicines and Medical Devices Agency of Serbia. Utilization was expressed as defined daily doses per 1000 inhabitants per day, and trends were assessed using linear and joinpoint regression analyses. Results: Total antipsychotic utilization increased from 4.35 to 14.42 DDD/1000 inhabitants/day, representing a 231.5% increase. This growth was predominantly driven by atypical antipsychotics, whose utilization increased from 1.16 to 10.91 DDD/1000 inhabitants/day (+840.2%). In contrast, typical antipsychotic utilization remained relatively stable in absolute terms. The share of atypical antipsychotics increased from 26.7% in 2006 to 75.6% in 2024, while the atypical: typical utilization ratio increased from 0.36 to 3.10. Atypical antipsychotics surpassed typical agents in 2013. Marked increases were observed for olanzapine, quetiapine, aripiprazole, paliperidone, risperidone, and clozapine, while chlorpromazine and fluphenazine declined. Conclusions: These findings demonstrate a substantial increase in overall antipsychotic utilization in Serbia and a pronounced structural shift toward atypical agents, highlighting the need for continued monitoring of prescribing trends, safety outcomes, and population-level treatment patterns. Full article

(This article belongs to the Section Pharmacology)

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19 pages, 9387 KB

Open AccessArticle

An Alkaloid from Marine Sirastachys pandanicola Inhibiting Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase Activity

by Yang Man, Zihao Wang, Boyu Chen, Xiaozhen Diao, Hideo Kigoshi, Yiwen Zhao, Jeevithan Elango, Ahsan Javed and Wenhui Wu

Pharmaceuticals 2026, 19(7), 1127; https://doi.org/10.3390/ph19071127 - 21 Jul 2026

Viewed by 332

Abstract

Background/Objectives: Marine microorganism metabolites are structurally unique secondary metabolites possessing therapeutic potential. The current study aims to identify a novel ATPase regulator using a newly established bidirectional activity evaluation system to screen for microbial metabolites that inhibit the activities of Na⁺ [...] Read more.

Background/Objectives: Marine microorganism metabolites are structurally unique secondary metabolites possessing therapeutic potential. The current study aims to identify a novel ATPase regulator using a newly established bidirectional activity evaluation system to screen for microbial metabolites that inhibit the activities of Na⁺-K⁺-ATPase or Ca²⁺-Mg²⁺-ATPase. Methods: A total of 1258 marine microbial strains were isolated from sea mud in Zhoushan, Zhejiang. Results: The extract of strain ZSDH2536 exhibited Na⁺-K⁺ and Ca²⁺-Mg²⁺-ATPase inhibitory activity and was identified as Sirastachys pandanicola based on morphological and molecular phylogenetic analyses. The secondary metabolite was tentatively identified in the ZSDH2536 strain as a bisindole compound, and named Pandanicoline based on ¹H-NMR, ¹³C-NMR and high-resolution mass spectrometry analysis. The chemical formula of Pandanicoline is C₅₁H₆₈N₂O₁₀, with an isotopic mass of 868.4874 Da. The maximum inhibition rate of Pandanicoline on Na⁺-K⁺ and Ca²⁺-Mg²⁺-ATPase was 36.37% and 37.27%, respectively. Moreover, in silico analysis also showed the binding energy of Pandanicoline with Na⁺-K⁺-ATPase was −9.124 kcal/mol and with the Ca²⁺-Mg²⁺-ATPase complex was −10.47 kcal/mol. Conclusions: The strain ZSDH2536 represents a promising source of dual inhibitors targeting Na⁺-K⁺ and Ca²⁺-Mg²⁺-ATPase. Pandanicoline exhibits potential as a lead compound for regulating ion homeostasis, providing new opportunities for further investigation into its mechanism and therapeutic applications. Full article

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32 pages, 5570 KB

Open AccessArticle

Computational Evaluation of Multitarget Capabilities of Phenylethanoid Glycosides Against SARS-CoV-2’s 3CL^pro and PL^pro

by Maria Eduarda Alves Esteves, Bruce Veiga Andriolo, Caio Felipe de Araujo Ribas Cheohen, Thamirys Silva da Fonseca, Mariana Freire Campos, Carla Monteiro Leal, Diego Allonso, Gilda Guimarães Leitão, Suzana Guimarães Leitão and Manuela Leal da Silva

Pharmaceuticals 2026, 19(7), 1126; https://doi.org/10.3390/ph19071126 - 21 Jul 2026

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Abstract

Background: The development of multitarget drugs capable of simultaneously inhibiting SARS-CoV-2 proteases—3CL^pro and PL^pro—may enhance therapeutic efficacy against COVID-19. Given the historical use of Traditional Chinese Medicine (TCM) in the management of respiratory diseases, phenylethanoid glycosides (PGs) represent an attractive [...] Read more.

Background: The development of multitarget drugs capable of simultaneously inhibiting SARS-CoV-2 proteases—3CL^pro and PL^pro—may enhance therapeutic efficacy against COVID-19. Given the historical use of Traditional Chinese Medicine (TCM) in the management of respiratory diseases, phenylethanoid glycosides (PGs) represent an attractive and chemically diverse natural product scaffold for the discovery of antiviral agents. Objectives: This study aimed to identify promising candidates within this class capable of simultaneously inhibiting both target proteases. Methods: The PG structures described in the literature between 1950 and 2020 were gathered and curated to construct a dedicated database, which was subsequently subjected to virtual screening. In silico ADMETox predictions and 2D ligand–protein interaction analyses were then employed to evaluate the identified hit PGs. Ligand stability within the binding sites of the proteases was further assessed using free energy landscape (FEL) and MM/GBSA calculations, while enzymatic inhibition of the commercial PG was evaluated via FRET assays. Results: Virtual screening identified 22 PGs with multitarget potential, predominantly sourced from Asia, followed by the Americas and Europe. The hit compound magnoloside I is found in Magnolia officinalis, a species widely used in TCM for respiratory conditions and officially prescribed during the COVID-19 pandemic. A second hit, calceolarioside B, inhibited more than 90% of the enzymatic activity of both proteases in the FRET assay. Conclusions: Together, these findings highlight phenylethanoid glycosides as promising scaffolds for dual protease inhibition. Full article

(This article belongs to the Section Medicinal Chemistry)

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21 pages, 5178 KB

Open AccessArticle

L-Cysteine Ethyl Ester May Overcome Morphine-Induced Respiratory Depression by Activating Muscarinic Receptors

by Paulina M. Getsy, Walter J. May, Santhosh M. Baby, Gregory A. Coffee, Hubert V. Forster, Matthew R. Hodges, Yunguang Qiu, Feixiong Cheng, James N. Bates and Stephen J. Lewis

Pharmaceuticals 2026, 19(7), 1125; https://doi.org/10.3390/ph19071125 - 21 Jul 2026

Cited by 1 | Viewed by 340

Abstract

Background/Objectives: Opioids inhibit breathing that can lead to fatal overdose, highlighting the need for testing effective countermeasure agents and potential mechanisms of action. Here we examined the role muscarinic cholinergic receptors play in the ability of L-cysteine ethyl ester (L-CYSee) to overcome the [...] Read more.

Background/Objectives: Opioids inhibit breathing that can lead to fatal overdose, highlighting the need for testing effective countermeasure agents and potential mechanisms of action. Here we examined the role muscarinic cholinergic receptors play in the ability of L-cysteine ethyl ester (L-CYSee) to overcome the deleterious effects of morphine on ventilatory parameters in male Sprague Dawley rats and the ventilatory responses during a subsequent hypoxic-hypercapnic (HH) challenge. Methods: Ventilatory parameters were measured by whole body plethysmography. Results: The injection of the muscarinic receptor antagonist, atropine (1.0 mg/kg, IV), elicited an array of ventilatory responses (e.g., an increase in frequency of breathing coupled with a fall in tidal volume). Injection of morphine (10 mg/kg, IV) to vehicle-treated rats elicited a depression of breathing, including sustained decreases in tidal volume, minute ventilation, peak inspiratory flow, and peak inspiratory and expiratory drives, which were associated with marked increases in end inspiratory pause (EIP) and end expiratory pause (EEP), expiratory flow at 50% expired tidal volume (EF₅₀), and rate of achieving peak expiratory flow (Rpef). Most effects of morphine (10 mg/kg, IV) were not altered in atropine-treated rats, except that increases in EIP, EEP and Rpef were reduced. Subsequent injections of L-CYSee (2 × 500 μmol/kg, IV given 15 min apart) overcame the adverse actions of morphine on ventilatory parameters in vehicle-treated rats. The effects of L-CYSee, such as reversal of the effects of morphine on frequency of breathing, tidal volume and minute ventilation, were markedly reduced in atropine-treated rats. The ability of L-CYSee to reverse the adverse effects of morphine to a HH gas challenge was markedly diminished in atropine-treated rats. Conclusions: These findings demonstrate that muscarinic receptors play a vital role in the ability of L-CYSee to overcome the deleterious effects of morphine. Full article

(This article belongs to the Section Pharmacology)

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36 pages, 518 KB

Open AccessReview

Use of Antihistamine Drugs in Colitis: A Review

by Bartosz Bogielski, Dariusz Gach, Katarzyna Michalczyk, Bronisława Skrzep-Poloczek, Mateusz Stojko, Jolanta Zalejska-Fiolka and Dominika Stygar

Pharmaceuticals 2026, 19(7), 1124; https://doi.org/10.3390/ph19071124 - 21 Jul 2026

Viewed by 559

Abstract

Background/Objectives: Colitis involves both local intestinal damage and systemic dysfunction, driven by oxidative stress and immune imbalance. Histamine, acting through its receptors, influences these processes, yet its therapeutic relevance in colitis remains unclear. This review systematically examines histamine-mediated signaling in colitic inflammation [...] Read more.

Background/Objectives: Colitis involves both local intestinal damage and systemic dysfunction, driven by oxidative stress and immune imbalance. Histamine, acting through its receptors, influences these processes, yet its therapeutic relevance in colitis remains unclear. This review systematically examines histamine-mediated signaling in colitic inflammation and evaluates preclinical and clinical data on antihistamines to assess their potential as a mechanism-based treatment. Methods: A structured literature search was conducted using PubMed and Google Scholar to identify studies addressing histamine signaling and the use of antihistamines in colitis, using keywords such as “colitis,” “histamine,” “histamine receptors,” and “antihistamines.” Relevant experimental and clinical studies were screened and critically analyzed to provide an integrated overview of mechanistic and therapeutic insights. This review was designed as a mechanistic narrative synthesis based on a structured search and qualitative appraisal rather than a formal systematic review with quantitative risk-of-bias grading. Results: Findings from preclinical investigations consistently indicate that pharmacological manipulation of histamine signaling—especially through H3 and H4 receptor subtypes—reduces inflammatory activity and modulates oxidative balance in animal models of colitis. Conversely, clinical evidence concerning H1 and H2 receptor antagonists remains scarce and discordant, lacking sufficient support for a definitive causal relationship or unambiguous therapeutic efficacy. Importantly, no clinical studies to date have assessed the effects of selective H3 or H4 receptor blockers in individuals with colitis, revealing a substantial disconnect between bench research and bedside application. Existing human trials have mainly concentrated on mucosal endpoints, with little attention paid to systemic manifestations or oxidative stress-related parameters. Conclusions: Histamine-dependent signaling constitutes a mechanistically credible target in colitis, connecting immune activation, impairment of the epithelial barrier, and oxidative injury. Although experimental data are encouraging, clinical corroboration remains absent. Antihistamines may hold greater promise for alleviating systemic oxidative stress than for directly ameliorating intestinal inflammation. Rigorously designed clinical trials that incorporate both gastrointestinal and systemic outcome measures are necessary to elucidate their therapeutic position. Full article

(This article belongs to the Special Issue Recent Advances in the Discovery and Development of Drugs for Civilization Diseases, 2nd Edition)

34 pages, 25538 KB

Open AccessArticle

A Deep Learning Framework for the Discovery of Natural-Product Candidate Binders of Acetyl-CoA Carboxylase 2 (ACC2) with Potential Relevance to Cardiometabolic Lipid Metabolism

by Nada A. Alzunaidy

Pharmaceuticals 2026, 19(7), 1123; https://doi.org/10.3390/ph19071123 - 21 Jul 2026

Viewed by 390

Abstract

Background/Objectives: Obesity and related metabolic diseases arise from an interplay of lipid overload, insulin resistance and oxidative stress. Acetyl-CoA carboxylase 2 (ACC2) controls malonyl-CoA production and thereby gates mitochondrial fatty-acid oxidation, placing it at the intersection of lipid handling and redox-sensitive metabolic dysfunction. [...] Read more.

Background/Objectives: Obesity and related metabolic diseases arise from an interplay of lipid overload, insulin resistance and oxidative stress. Acetyl-CoA carboxylase 2 (ACC2) controls malonyl-CoA production and thereby gates mitochondrial fatty-acid oxidation, placing it at the intersection of lipid handling and redox-sensitive metabolic dysfunction. Dietary antioxidants such as polyphenols, flavonoids and terpenoids are increasingly studied as modulators of these pathways, yet systematic prioritization of food-derived antioxidant compounds against defined metabolic targets remains challenging. We developed an integrated deep learning and structure-based workflow to prioritize FooDB compounds with predicted ACC2-binding potential. Methods: A curated set of 3983 ACC2 bioactivity records from ChEMBL 36 was used to train scaffold-split models, including graph neural-network and graph–Morgan fingerprint-fusion architectures. The calibrated ensemble screened 139,988 FooDB compounds; 200 candidates with predicted activity probability above 0.70 were docked against the ACC2 carboxyltransferase domain (PDB ID: 3FF6), and six prioritized complexes underwent 500 ns molecular dynamics and MM/GBSA analysis. Results: Redocking of the co-crystallized ligand reproduced the experimental pose (RMSD 1.2 Å). Although the highest-ranked screening hits were antioxidant terpenoids and alkaloids, docking-based prioritization from the top candidates selected six larger, more polar food-derived compounds, including glycosides and two nucleotide/cofactor-like conjugates, which showed docking scores from −7.47 to −6.65 kcal/mol versus −6.21 kcal/mol for the reference ligand. Glu539 emerged as a recurrent interaction hotspot. All candidates gave more favourable MM/GBSA binding free energies than the reference (ΔG = −22.52 kcal/mol), led by FDB029596 (−35.65), FDB021568 (−34.14) and FDB017807 (−33.87 kcal/mol). Conclusions: This workflow provides a reproducible framework for prioritizing food-derived compounds as candidate ACC2 binders relevant to obesity and metabolic disease, generating structurally supported hypotheses for biochemical and nutritional validation. The prioritized compounds are computational candidates only and require biochemical and cellular (experimental) validation before any ACC2-related biological relevance can be established. Full article

(This article belongs to the Section AI in Drug Development)

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19 pages, 1089 KB

Open AccessArticle

Hepatic Safety Profile of Atomoxetine and Methylphenidate in Patients with ADHD: Disproportionality Analysis Using EudraVigilance Database Data

by Raffaella Di Napoli, Ludovica Vittoria Laino, Concetta Rafaniello, Luigi Di Costanzo, Maria Giuseppa Sullo, Cristina Scavone and Annalisa Capuano

Pharmaceuticals 2026, 19(7), 1122; https://doi.org/10.3390/ph19071122 - 21 Jul 2026

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Abstract

Background: Hepatotoxicity induced by atomoxetine (ATX) and methylphenidate (MPH) when used to treat ADHD is a rare but potentially serious complication. This study aims to describe the hepatic adverse drug reactions (ADRs) reported for ATX and MPH by analysing data from the [...] Read more.

Background: Hepatotoxicity induced by atomoxetine (ATX) and methylphenidate (MPH) when used to treat ADHD is a rare but potentially serious complication. This study aims to describe the hepatic adverse drug reactions (ADRs) reported for ATX and MPH by analysing data from the EudraVigilance database. Methods: Individual case safety reports (ICSRs) listing ATX and/or MPH as suspected drugs and reporting at least one adverse event (AE) within the ‘hepatobiliary disorders’ system organ class (SOC) were extracted for the period from 1 January 2012 to 20 May 2025. Descriptive and disproportionality analyses were then performed. Results: During the study period, 421 ICSRs reporting AEs classified under the “hepatobiliary disorders” SOC and involving ATX and/or MPH as suspected drugs were retrieved (ATX, N = 232; MPH, N = 181). Most cases involved adult (N = 261) and female (N = 222) patients. The majority of reports were classified as serious (N = 349). Overall, 375 AEs were identified. Drug-induced liver injury (DILI) was the most frequently reported AE (N = 103 ATX; N = 47 MPH), followed by hepatitis (N = 20 ATX; N = 9 MPH) and jaundice (N = 15 ATX; N = 20 MPH). The disproportionality analysis, based on a head-to-head comparison, showed a higher reporting frequency of hepatobiliary disorders for ATX compared to MPH (ROR 2.41 [95%CI 2.11–3.11]). Specifically, ATX was associated with significantly higher reporting frequencies than MPH for the AEs of DILI, hepatitis, and jaundice (6.42 [4.45–9.06]; 6.48 [2.95–14.24]; and 2.19 [1.12–4.27], respectively). Conclusions: This analysis, based on the EudraVigilance database, suggests that both drugs are associated with hepatobiliary adverse drug reactions, with a higher overall reporting frequency observed for ATX. Full article

(This article belongs to the Special Issue Neuropsychiatric Disorders: Pharmacological Aspects)

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30 pages, 2794 KB

Open AccessArticle

Bacterial Etiology and Antimicrobial Susceptibility Patterns in Pediatric Intra-Abdominal Infections—Implications for Empirical Treatment and Antimicrobial Stewardship

by Florin Daniel Enache, Ancuta Lupu, Tatiana Chisnoiu, Adriana Luminita Balasa, Emil Anton, Gabriel Florin Panculescu, Ioana Livia Suliman, Violeta Popovici, Ramona Mihaela Stoicescu, Iulian Manac, Vasile Valeriu Lupu and Cristina Maria Mihai

Pharmaceuticals 2026, 19(7), 1121; https://doi.org/10.3390/ph19071121 - 20 Jul 2026

Viewed by 1143

Abstract

Objectives: Intra-abdominal infections (IAIs) requiring surgical intervention represent a significant cause of morbidity in pediatric patients, often leading to prolonged hospitalization and increased antimicrobial exposure. This study aimed to characterize the distribution of pathogenic bacteria, antimicrobial resistance patterns, and clinical associations in [...] Read more.

Objectives: Intra-abdominal infections (IAIs) requiring surgical intervention represent a significant cause of morbidity in pediatric patients, often leading to prolonged hospitalization and increased antimicrobial exposure. This study aimed to characterize the distribution of pathogenic bacteria, antimicrobial resistance patterns, and clinical associations in pediatric intra-abdominal infections complicated by surgical site involvement. Methods: A retrospective observational study was conducted on children aged 0–16 years, who underwent surgery for intra-abdominal infections with microbiological confirmation. Peritoneal fluid, pus, and other intraoperative or postoperative specimens were analyzed using standard microbiological techniques. Infections were classified as monomicrobial or polymicrobial, and antimicrobial susceptibility was assessed phenotypically. Associations between bacterial pathogens, patient age, underlying diagnosis, surgical procedures, and antibiotic susceptibility patterns were analyzed. Results: 177 pediatric patients were included. Appendicitis was the most common diagnosis (53.53%), followed by intra-abdominal abscesses (32.94%). Gram-negative (GN) bacteria predominated (44.07%), with Escherichia coli being the most frequently isolated pathogen (83.33%). Double and triple GN and GP associations were identified in approximately 25% of cases, particularly in abscesses, complicated appendicitis, and surgical site infections. Our findings revealed that piperacillin–tazobactam and carbapenems were expected to be effective against almost all GN pathogens identified in surgical specimens of pediatric patients. Pathogen distribution and antimicrobial susceptibility varied significantly according to age group and clinical diagnosis. Conclusions: In pediatric patients, intra-abdominal infections requiring surgical management were mainly caused by Gram-negative bacteria and polymicrobial associations. Effective treatment relies on prompt surgical source control and empiric broad-spectrum antimicrobial therapy, followed by culture-guided de-escalation to support antimicrobial stewardship. Full article

(This article belongs to the Special Issue Epidemiological Surveillance of Clinical Bacterial Infections and Antimicrobial Resistance Management)

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28 pages, 20646 KB

Open AccessArticle

Machine Learning and Molecular Modeling Strategy for the Identification of CNS-Active Acetylcholinesterase Inhibitors

by Muhammad Yasir, Jinyoung Park, Eun-Taek Han, Won Sun Park, Jin-Hee Han, Jongseon Choe and Wanjoo Chun

Pharmaceuticals 2026, 19(7), 1120; https://doi.org/10.3390/ph19071120 - 20 Jul 2026

Viewed by 321

Abstract

Background: Acetylcholinesterase (AChE) is a key therapeutic target in neurological disorders, and the discovery of novel inhibitors with improved efficacy and pharmacokinetic properties remains a significant challenge. Methods: In this study, an integrated computational and experimental approach was employed to identify potential AChE [...] Read more.

Background: Acetylcholinesterase (AChE) is a key therapeutic target in neurological disorders, and the discovery of novel inhibitors with improved efficacy and pharmacokinetic properties remains a significant challenge. Methods: In this study, an integrated computational and experimental approach was employed to identify potential AChE inhibitors. A machine learning-based model was developed to predict bioactive compounds from large chemical libraries, followed by Blood–Brain Barrier (BBB) permeability screening to ensure Central Nervous System (CNS) suitability. The shortlisted compounds were further evaluated using molecular docking, molecular dynamics simulations, and MM-PBSA binding free-energy calculations to assess their interaction profiles and stability. Selected top-ranked compounds were subjected to in vitro biological evaluation for AChE inhibitory activity. Results: The results demonstrated that several screened compounds, including Z1498348710 and Z1824281875, exhibited notable inhibition with AChE activity reduced to approximately 75% and 72%, respectively, compared to the control. Other compounds such as Z29542160, Z105150208, Z94570687, and Z94570675 showed moderate inhibitory effects, maintaining AChE activity in the range of 82–86%. In comparison, the reference inhibitors Donepezil and Neostigmine bromide displayed significantly stronger inhibition, reducing AChE activity to approximately 20% and 18%, respectively. Conclusions: Overall, the identified compounds demonstrated moderate AChE inhibitory activity while exhibiting favorable predicted physicochemical and computational profiles. These findings suggest that they represent promising starting points for medicinal chemistry optimization and future development as CNS-active AChE inhibitors. Full article

(This article belongs to the Special Issue QSAR and Chemoinformatics in Drug Design and Discovery)

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17 pages, 7090 KB

Open AccessArticle

Preclinical Evaluation of Dual NOP/Mu Partial Agonists for Methamphetamine Use Disorder

by Thamires Righi, Gilles Zribi, Sebastian Garcia, Stephen M. Husbands, Lawrence Toll and Andrea Cippitelli

Pharmaceuticals 2026, 19(7), 1119; https://doi.org/10.3390/ph19071119 - 20 Jul 2026

Viewed by 286

Abstract

Background/Objectives: Methamphetamine (meth) use disorder (MUD) continues to pose a significant public health challenge, and there are currently no FDA-approved pharmacological treatments available. Compounds that simultaneously activate nociceptin opioid peptide (NOP) and mu opioid receptors offer a promising therapeutic approach for substance use [...] Read more.

Background/Objectives: Methamphetamine (meth) use disorder (MUD) continues to pose a significant public health challenge, and there are currently no FDA-approved pharmacological treatments available. Compounds that simultaneously activate nociceptin opioid peptide (NOP) and mu opioid receptors offer a promising therapeutic approach for substance use disorders, including psychostimulant addiction. Methods: We evaluated two mixed NOP/mu receptor agonists for their ability to reduce meth intake using a translational drug-versus-food choice self-administration paradigm in male and female Sprague–Dawley rats. In addition, both compounds were tested in female rats for their effects on cue- and priming-induced reinstatement of meth-seeking behavior, established preclinical models of relapse. Results: PPL-138 and PPL-143 are structurally related bifunctional NOP/mu partial agonists that exhibit greater efficacy at NOP than at mu receptors, with PPL-143 showing the highest NOP efficacy. In behavioral assays, both compounds significantly reduced meth self-administration in male rats and exhibited comparable potency in this effect. In female rats, however, PPL-143 was less effective than PPL-138 in reducing meth intake. In reinstatement models, PPL-138, but not PPL-143, attenuated prime-induced reinstatement of meth seeking, whereas both compounds produced comparable increases in meth seeking in the cue-induced reinstatement paradigm. Additional findings indicated that PPL-138 enhanced meth-induced locomotor activation. Conclusions: The findings support mixed mu/NOP partial agonists as a promising pharmacological strategy for treating MUD, while also indicating a potential for increased drug seeking under certain conditions. Notably, enhancing the NOP component does not appear to meaningfully improve either the safety profile or the anti-meth efficacy of these bifunctional ligands. Full article

(This article belongs to the Section Pharmacology)

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17 pages, 1597 KB

Open AccessArticle

Antidepressants and Road Safety: A Forensic Toxicological Perspective Based on Observational Data and Current Evidence

by Davide Filardi, Francesca Vernich, Federico Mineo, Giulio Mannocchi and Roberta Tittarelli

Pharmaceuticals 2026, 19(7), 1118; https://doi.org/10.3390/ph19071118 - 20 Jul 2026

Viewed by 383

Abstract

Background/Objectives: Antidepressants are widely prescribed medications that may affect psychomotor performance and driving ability depending on their pharmacological profile. This study investigated the prevalence and patterns of antidepressant use among drivers undergoing forensic toxicological evaluation and explored their potential implications for road safety. [...] Read more.

Background/Objectives: Antidepressants are widely prescribed medications that may affect psychomotor performance and driving ability depending on their pharmacological profile. This study investigated the prevalence and patterns of antidepressant use among drivers undergoing forensic toxicological evaluation and explored their potential implications for road safety. Methods: An observational study was conducted on n = 6316 drivers undergoing forensic toxicological assessment following licence suspension for driving under the influence (DUI) of alcohol and/or drugs between January 2023 and December 2025. Reported antidepressants were classified into selective serotonin reuptake inhibitors (SSRIs), serotonin–norepinephrine reuptake inhibitors (SNRIs), serotonin antagonist and reuptake inhibitors (SARIs), tricyclic antidepressants (TCAs), norepinephrine–dopamine reuptake inhibitors (NDRIs), noradrenergic and specific serotonergic antidepressants (NaSSAs), and monoamine oxidase inhibitors (MAOIs). Distribution patterns were analysed descriptively and discussed considering the available literature. Results: Antidepressant use was reported by n = 132 participants (2.1%). SSRIs were the most common class (44.0%), followed by SNRIs (26.5%) and SARIs (17.4%). TCAs (6.1%), NDRIs (3.7%), and NaSSAs (2.3%) were less common, while no MAOI use was reported. Among antidepressant users, n = 28 individuals (21.2%) tested positive for other psychoactive substances, including benzodiazepines, cocaine, and cannabinoids. Conclusions: The use of antidepressants was relatively uncommon in the study population. However, clinical evaluation remains important, particularly at the start of treatment and during dose adjustments to determine the potential for an increased risk while driving. Further studies integrating toxicological analyses and clinical data are needed to better define the relationship between antidepressant exposure, polysubstance use, and road safety. Full article

(This article belongs to the Special Issue Effects of Drug Abuse and Its Consequences on Health)

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19 pages, 1022 KB

Open AccessSystematic Review

Geranylgeraniol as a Modulator of Mevalonate Pathway Disruption: A Scoping Review of Cellular Mechanisms and Skeletal Outcomes in Osteoporosis Models

by Sophia Ogechi Ekeuku, Mohammed Farhan Abed Al Salman, Nur Vaizura Mohamad, Sok Kuan Wong and Kok-Yong Chin

Pharmaceuticals 2026, 19(7), 1117; https://doi.org/10.3390/ph19071117 - 20 Jul 2026

Viewed by 327

Abstract

Background/Objectives: Geranylgeraniol (GGOH), an isoprenoid intermediate of the mevalonate pathway, regulates bone cell viability and function, particularly by mitigating cellular toxicity induced by nitrogen-containing bisphosphonates (N-BPs). Despite this, its role in osteoporosis remains underexplored. This scoping review synthesises evidence on the effects of [...] Read more.

Background/Objectives: Geranylgeraniol (GGOH), an isoprenoid intermediate of the mevalonate pathway, regulates bone cell viability and function, particularly by mitigating cellular toxicity induced by nitrogen-containing bisphosphonates (N-BPs). Despite this, its role in osteoporosis remains underexplored. This scoping review synthesises evidence on the effects of GGOH in in vitro and in vivo models of osteoporosis. Methods: PubMed, Scopus, and Ovid were searched using GGOH- and osteoporosis-related terms. Primary studies evaluating GGOH exposure in cellular or animal osteoporosis models were eligible. Twenty-nine studies met the inclusion criteria. Results: In vitro findings demonstrate that GGOH reverses N-BP-induced depletion of geranylgeranyl pyrophosphate, restoring protein prenylation which is essential for osteoclast and osteoblast survival, cytoskeletal organisation, and differentiation. GGOH reduced osteoclast apoptosis, restored nuclear factor of activated T-cells 1 and carbonic anhydrase II expression, and prevented N-BP-associated suppression of bone resorption. In osteoblasts and mesenchymal stem cells, GGOH improved viability, upregulated osteogenic markers including runt-related transcription factor 2, alkaline phosphatase, collagen type I, and bone morphogenetic proteins, and rescued mineralisation impaired by alendronate or zoledronate. Independent of N-BPs, GGOH exerted divergent effects on osteoclasts, by inhibiting osteoclastogenesis or promoting retinoic acid receptor-mediated bone resorption and attenuating zoledronate protection in vascular calcification settings in a model-specific manner. In vivo, dietary GGOH supplementation improved trabecular and cortical bone parameters and reduced serum C-terminal telopeptide of type I collagen in obese mice, indicating suppression of bone resorption. Conclusions: Overall, although GGOH shows osteoprotective potential, its capacity to antagonise N-BP efficacy limits systemic co-administration. Current evidence suggests that local delivery may warrant future investigation as a strategy to mitigate N-BP-induced skeletal toxicity. However, studies evaluating bone tissue exposure, pharmacokinetics, and clinically achievable concentrations are required before this approach can be translated. Full article

(This article belongs to the Special Issue Aging-Related Musculoskeletal Disorders: Therapeutic Targets and Pharmacological Strategies)

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29 pages, 4589 KB

Open AccessReview

Preclinical Models of Bladder Cancer: Barrier, Metabolic, and Translational Susceptibility

by Tianjia Liu, Wei Li, Qinzhamusu Yin, Da Liu, Yong Wang and Ning Cui

Pharmaceuticals 2026, 19(7), 1116; https://doi.org/10.3390/ph19071116 - 20 Jul 2026

Viewed by 427

Abstract

Preclinical bladder cancer models are often judged by tumor take, tumor growth or treatment inhibition, yet these endpoints do not reveal which bladder-specific constraints a given model preserves or bypasses. The bladder is shaped by a specialized urothelial barrier, urine exposure, cyclic filling [...] Read more.

Preclinical bladder cancer models are often judged by tumor take, tumor growth or treatment inhibition, yet these endpoints do not reveal which bladder-specific constraints a given model preserves or bypasses. The bladder is shaped by a specialized urothelial barrier, urine exposure, cyclic filling and emptying, inflammatory injury, metabolic stress and intravesical treatment pressure. In this review, we use susceptibility engineering as an organizing framework for model selection and validation. We define susceptibility engineering as the deliberate definition, perturbation and reporting of model states that alter tumor initiation, adhesion, colonization, survival or therapeutic exposure. This framework groups cell lines, patient-derived organoids, cell-line-derived xenograft (CDX) and patient-derived xenograft (PDX) models, orthotopic transplantation, N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced tumors, genetically engineered mouse models and large-animal platforms according to the biological constraints they test. We focus on three linked dimensions: urothelial barrier integrity and uroplakin-related tools; local colonization thresholds under bladder-specific selection; metabolic susceptibility involving peroxisome proliferator-activated receptor gamma (PPARG)-associated differentiation programs and candidate solute carrier family 25 (SLC25)-linked mitochondrial stress nodes. We further distinguish large-animal systems as platforms for local delivery, imaging, device testing and procedural scale rather than universal substitutes for mouse models. A susceptibility-based validation framework could improve model selection, explain divergent responses across systems and support tiered platforms that connect patient-derived biology, mechanistic mouse studies and clinically realistic intravesical evaluation. Full article

(This article belongs to the Special Issue Animal Models in Pharmacological Research: Focusing on Oncology and Related Fields)

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40 pages, 4136 KB

Open AccessReview

From Whole-Plant Phytochemistry to Precision Oncology: A Paradigm-Shifting Systematic Review of Lycium barbarum L. (Goji Berries)

by Yuanhong Lan, Lanfei Ma, Yuxin Kong, Dina Mahemuti, Congcong Zhang, Yuxiang Zhang, Wenfang Li and Ayitila Maimaitijiang

Pharmaceuticals 2026, 19(7), 1115; https://doi.org/10.3390/ph19071115 - 20 Jul 2026

Viewed by 477

Abstract

Medicinal and edible plants are promising resources for low-toxicity therapeutics and precision nutrition. Lycium barbarum L. (goji berry), a quintessential medicine–food homologous herb with 2000 years of ethnopharmacological use, has attracted global attention. However, existing studies are limited by fruit-centric bias, lack of [...] Read more.

Medicinal and edible plants are promising resources for low-toxicity therapeutics and precision nutrition. Lycium barbarum L. (goji berry), a quintessential medicine–food homologous herb with 2000 years of ethnopharmacological use, has attracted global attention. However, existing studies are limited by fruit-centric bias, lack of correlation between processing methods, component properties and bioactivities, and incomplete antitumor mechanistic understanding. This systematic review establishes a holistic research paradigm that integrates whole-plant resource utilization, processing–property–bioactivity associations, multi-target pharmacology, and clinical translation. We delineate tissue-specific distributions of core bioactives (polysaccharides, phenolics, carotenoids, alkaloids) and their synergistic networks underlying antioxidant, anti-inflammatory, hypoglycemic, and immunomodulatory effects. Critically, we systematically summarize preclinical evidence for the antitumor potential of goji berries, identifying four proposed non-overlapping cell death pathways (apoptosis, cell cycle arrest, ferroptosis, autophagy) and proposed unique roles in reversing multi-drug resistance, alleviating chemoradiotherapy toxicity, and serving as biocompatible nanocarriers, all of which remain predominantly at the preclinical stage. We further propose a three-stage evidence-based roadmap to address key translational bottlenecks. This review bridges the gap between traditional ethnopharmacology and modern precision nutrition, providing a scientific foundation for the sustainable development of the global goji berry industry. Full article

(This article belongs to the Section Natural Products)

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12 pages, 1155 KB

Open AccessArticle

Selective CB₂ Agonist JWH-133 Suppresses Viability and Migration-Related Responses in Prostate Cancer Cells

by Seda Sabah Özcan, Rehime Yapar, İsmail Değerli, Levent Elmas, Mehmet Korkmaz and Murat Çakır

Pharmaceuticals 2026, 19(7), 1114; https://doi.org/10.3390/ph19071114 - 19 Jul 2026

Viewed by 413

Abstract

Background/Objectives: Cannabinoid receptor type 2 (CB₂) agonists have attracted attention because of their potential effects on tumor-related cellular processes in different cancer models. In the present study, we investigated the effects of the selective CB₂ agonist JWH-133 on prostate [...] Read more.

Background/Objectives: Cannabinoid receptor type 2 (CB₂) agonists have attracted attention because of their potential effects on tumor-related cellular processes in different cancer models. In the present study, we investigated the effects of the selective CB₂ agonist JWH-133 on prostate cancer cell lines (LNCaP, DU-145, and PC3). Methods: Cell viability was evaluated using the MTT assay, while colony-forming capacity and migration-related responses were assessed by colony formation and wound-healing assays, respectively. In addition, the expression levels of selected cell-cycle- and apoptosis-related genes were analyzed by quantitative real-time PCR. Results: JWH-133 reduced cell viability in a time- and concentration-dependent manner, although the magnitude of this effect differed among prostate cancer cell lines. The compound also reduced colony formation in PC3 and LNCaP cells and decreased wound closure in PC3 cells under the experimental conditions used. Furthermore, JWH-133 altered the expression of several cell-cycle- and apoptosis-related genes in DU-145 and PC3 cells. Conclusions: Overall, these findings suggest that JWH-133 modulates multiple cellular responses in prostate cancer cells in a cell-line-dependent manner. However, additional mechanistic studies are required to clarify the molecular pathways underlying these effects. Full article

(This article belongs to the Section Biopharmaceuticals)

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30 pages, 856 KB

Open AccessReview

Gut Microbiota and Metabolic Dysfunction-Associated Steatotic Liver Disease: From Dysbiosis to Metagenomic Insights and Therapeutic Perspectives

by Otilia Elena Frăsinariu, Violeta Ștreangă, Aniela Luminița Rugină, Dana Elena Mîndru, Teodora Cristina Vintilă, Oana Viola Bădulescu, Iris Bararu-Bojan, Vasile Valeriu Lupu, Ancuța Lupu, Adriana Mihai, Isabela Ioana Loghin, Daniela Eugenia Popescu and Dragoș Florin Teșoi

Pharmaceuticals 2026, 19(7), 1113; https://doi.org/10.3390/ph19071113 - 19 Jul 2026

Viewed by 507

Abstract

Metabolic dysfunction-associated steatotic liver disease (MASLD) has emerged as the most common chronic liver disorder in the pediatric population, closely paralleling the global rise in childhood obesity. Increasing evidence highlights the gut–liver axis as a key contributor to MASLD pathogenesis, with gut microbiota [...] Read more.

Metabolic dysfunction-associated steatotic liver disease (MASLD) has emerged as the most common chronic liver disorder in the pediatric population, closely paralleling the global rise in childhood obesity. Increasing evidence highlights the gut–liver axis as a key contributor to MASLD pathogenesis, with gut microbiota dysbiosis influencing hepatic steatosis through multiple interconnected mechanisms, including increased intestinal permeability, endotoxemia, altered bile acid metabolism, and modulation of host energy homeostasis. In children, the characterization of microbiota signatures associated with MASLD remains challenging due to heterogeneity across studies, age-related microbial dynamics, and methodological variability. This review synthesizes current evidence regarding the role of the gut microbiota in pediatric MASLD, focusing on pathogenetic pathways, reported microbial patterns, and microbiota-targeted therapeutic strategies, while incorporating relevant mechanistic evidence from adult studies where pediatric data remain limited. Although several taxa have been repeatedly associated with pediatric MASLD, findings are not yet sufficiently consistent for clinical application. Interventions such as probiotics, prebiotics, and dietary modulation show promising but still preliminary results, with limited high-quality pediatric trials available. A deeper mechanistic understanding and standardized study designs are needed to clarify causality and to support microbiota-based precision approaches in pediatric MASLD management. Full article

(This article belongs to the Special Issue The Regulatory Roles of the Gut Microbiota in Multisystem Diseases)

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25 pages, 1857 KB

Open AccessArticle

Cognitive and Mood Effects of a Soluble Mango Leaf Extract (Zynamite^® S): A Randomized, Double-Blind, Placebo-Controlled, Crossover Replication Trial

by Ana Beltrán-Arranz, Agustín Aibar-Almazán, David Fuentes-Ríos, Rubén Pérez-Machín, María del Carmen Carcelén-Fraile, Laura López-Ríos and Yolanda Castellote-Caballero

Pharmaceuticals 2026, 19(7), 1112; https://doi.org/10.3390/ph19071112 - 18 Jul 2026

Viewed by 615

Abstract

Background/Objectives: Botanical nootropics are increasingly sought as natural alternatives to synthetic stimulants. Mangifera indica leaf extract, standardized to the polyphenol mangiferin, has shown promise in regulating brain activity and enhancing cognitive performance. This study aimed to replicate, in an independent cohort, the acute [...] Read more.

Background/Objectives: Botanical nootropics are increasingly sought as natural alternatives to synthetic stimulants. Mangifera indica leaf extract, standardized to the polyphenol mangiferin, has shown promise in regulating brain activity and enhancing cognitive performance. This study aimed to replicate, in an independent cohort, the acute cognitive benefits of a soluble mango leaf extract (Zynamite^® S) following a previous proof-of-concept trial. Methods: In a double-blind, randomized, placebo-controlled crossover trial, 88 healthy young adults (aged 18–25) received either a single 100 mg dose of Zynamite^® S or a matched placebo. Cognitive performance was assessed using the Trail Making Test (TMT), Digit Symbol Substitution Test (DSST), and Stroop Color-Word Test. Emotional states were assessed via the Profile of Mood States (POMS) at baseline, and then at 30 min, 3 h, and 5 h post-ingestion. Results: Zynamite^® S supplementation resulted in the replication of previous findings, with significant improvements in mental processing speed, attention, and cognitive flexibility (p < 0.05). Notably, this trial identified a rapid onset of action, with significant cognitive and mood improvements detectable as early on as 30 min post-dose and a sustained duration of the effect with benefits observed up to 5 h post-administration. Mood assessments confirmed an overall improvement in emotional balance (p < 0.05) by reducing tension, mental fatigue, and low mood during the completion of cognitive-demanding tasks. Conclusions: These findings reproduce the acute benefits of Zynamite^® S in a healthy young adult population, including a rapid onset of effects and a sustained improvement in cognitive performance and mood over a 5 h window. Full article

(This article belongs to the Section Natural Products)

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15 pages, 19707 KB

Open AccessArticle

Casticin Alleviates Acetaminophen-Induced Acute Liver Injury by Modulating the TLR4/MyD88/TRAF6/NF-κB Signaling Pathway

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

Pharmaceuticals 2026, 19(7), 1111; https://doi.org/10.3390/ph19071111 - 18 Jul 2026

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Abstract

Background: Acute liver injury (ALI) is commonly caused by acetaminophen (APAP) overdose, which drives oxidative stress alongside activation of innate immune signaling. Casticin, a naturally occurring flavonoid, has anti-inflammatory and antioxidant properties. Focusing on the toll-like receptor 4 (TLR4)/myeloid differentiation primary response 88 [...] Read more.

Background: Acute liver injury (ALI) is commonly caused by acetaminophen (APAP) overdose, which drives oxidative stress alongside activation of innate immune signaling. Casticin, a naturally occurring flavonoid, has anti-inflammatory and antioxidant properties. Focusing on the toll-like receptor 4 (TLR4)/myeloid differentiation primary response 88 (MyD88)/tumor necrosis factor receptor-associated factor 6 (TRAF6)/nuclear factor kappa B (NF-κB) pathway, this study assessed casticin’s ability to protect mice from APAP-induced hepatotoxicity. Methods: APAP-induced ALI was established in mice randomly assigned to the following six groups: normal control, casticin control, APAP, APAP plus N-acetylcysteine (NAC), APAP plus low-dose casticin, and APAP plus high-dose casticin. Casticin was administered for three consecutive days before APAP to evaluate its preventive rather than therapeutic potential. Biochemical and histological analyses were performed, with molecular assessments using Western blotting, ELISA, quantitative real-time PCR (qPCR), and immunohistochemistry. Results: APAP significantly elevated serum ALT, AST, and ALP and markedly deteriorated hepatic architecture, confirming hepatotoxicity. APAP also induced lipid peroxidation and depleted antioxidant defenses. Hepatic TNF-α and IL-6 increased, IL-10 decreased, and the abundance of TLR4, MyD88, TRAF6, and NF-κB p65 was elevated. Casticin reduced these pathway components, lowered TNF-α and IL-6, and increased IL-10 dose-dependently, with effects approaching those of NAC. Conclusions: Casticin protected the liver against APAP toxicity, restraining oxidative injury while damping TLR4/MyD88/TRAF6/NF-κB signaling. Full article

(This article belongs to the Special Issue Natural Products Drug Discovery and Their Pharmacological Applications)

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15 pages, 5289 KB

Open AccessArticle

Portable, Rapid, and Cost-Effective Smartphone-Based Colorimetric Quantification of Total Lactones in Andrographis paniculata

by Sutasinee Apichai, Suphakorn Katib, Teerapat Ouirungroj, Thanawat Pattananandecha, Kanokwan Kiwfo, Fumihiko Ogata, Naohito Kawasaki, Kate Grudpan and Chalermpong Saenjum

Pharmaceuticals 2026, 19(7), 1110; https://doi.org/10.3390/ph19071110 - 18 Jul 2026

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Abstract

Background/Objectives: Andrographis paniculata is listed in the Thai National List of Essential Medicines. The Thai Herbal Pharmacopoeia specifies the required contents of total lactones and andrographolide for the quality control of the aerial parts of A. paniculata. Effective pre-harvest quality control [...] Read more.

Background/Objectives: Andrographis paniculata is listed in the Thai National List of Essential Medicines. The Thai Herbal Pharmacopoeia specifies the required contents of total lactones and andrographolide for the quality control of the aerial parts of A. paniculata. Effective pre-harvest quality control throughout the cultivation period is essential to ensure compliance with these quality standards. In this study, we aimed to develop a portable smartphone-based colorimetric method for the determination of total lactone content, thereby facilitating in-field quality control of A. paniculata raw materials. Methods: Methanol was used as the extraction solvent, and the analytes were extracted using a simple procedure derived from the United States Pharmacopeia concept. The colorimetric reaction was based on a charge-transfer reaction between the α,β-unsaturated γ-lactone moiety and 3,5-dinitrobenzoic acid, producing a red-purple product in a microwell plate. The delta-green intensity of the reaction product was captured as an andrographolide equivalent using a smartphone camera and quantified through digital image processing with a custom-developed mobile application. Results: A linear working range of 15–100 µg/mL was exhibited with limits of detection and quantification of 4.9 and 15.0 µg/mL, respectively. The practical applicability of the developed method was evaluated using A. paniculata samples and compared with the conventional spectrophotometric method. The results showed excellent agreement between the two methods, with a correlation coefficient of 0.9986. Conclusions: The results of this study suggest that this portable and rapid method is feasible for in-field/on-site analysis to facilitate pre-harvest quality control throughout the cultivation period, ensuring that harvested A. paniculata materials comply with established quality standards for plant-derived natural active pharmaceutical ingredients (NAPIs). The proposed method has the potential to promote sustainable production and responsible resource utilization by improving the quality of herbal raw materials intended for the manufacture of herbal medicines and dietary supplements, in line with UN-SDG #12 and #3. Full article

(This article belongs to the Special Issue Quality Control and Standardization of Plant-Based Pharmaceuticals)

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15 pages, 1572 KB

Open AccessArticle

Real-World Comparison of Trough- and Online-Bayesian-Calculator-Derived AUC_0–24/MIC Target Attainment for Vancomycin Monitoring in Critically Ill Patients

by Sufyan Alomair, Zahra Alsultan, Fatimah Alsultan, Fatimah Alghadeer, Alzahraa Aljafar, Anas Alkhawaldeh, Ayat Alherz and Batool Alhassan

Pharmaceuticals 2026, 19(7), 1109; https://doi.org/10.3390/ph19071109 - 18 Jul 2026

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Abstract

Introduction: The preferred PK/PD goal for vancomycin is now AUC_0–24/MIC-guided dosing, which requires two blood samples, additional time, increased cost, and possibly specialized staff, potentially limiting its use. Our study compared vancomycin target attainment using trough versus single-sample Bayesian AUC_0–24 [...] Read more.

Introduction: The preferred PK/PD goal for vancomycin is now AUC_0–24/MIC-guided dosing, which requires two blood samples, additional time, increased cost, and possibly specialized staff, potentially limiting its use. Our study compared vancomycin target attainment using trough versus single-sample Bayesian AUC_0–24/MIC in critically ill patients, using a validated online calculator. Methods: This retrospective cohort study included adults aged ≥18 years with stable renal function who were receiving vancomycin. Exclusions were age under 18, unstable renal function, hemodialysis, missing data, or non-steady-state vancomycin. Steady-state troughs were obtained, and AUC_0–24/MIC was calculated using ClinCalc. The primary endpoint was discordance in target attainment between trough (15–20 mg/L) and AUC_0–24/MIC (400–600 mg·h/L). The secondary endpoint was AKI within 48 h of initiating vancomycin. Results: The mean trough was 14.8 ± 8.0 mg/L, and the mean AUC_0–24/MIC was 499.7 ± 201.3 mg·h/L. Trough levels were within target in 20%, and AUC_0–24/MIC in 67%. The 3 × 3 cross-tabulation showed a significant association (χ² = 27.33, p < 0.001), but only fair agreement (Cohen’s κ = 0.215) and a biased disagreement pattern (p < 0.001). Thirty-eight patients (38%) had “false alarming” results: trough < 15 mg/L despite AUC_0–24/MIC ≥ 400 mg·h/L. Three patients (3.0%) showed “falsely reassuring” results: trough 15–20 mg/L with AUC_0–24/MIC > 600 mg·h/L. The odds of supratherapeutic AUC_0–24/MIC > 600 mg·h/L were tenfold higher in patients with trough 15–20 mg/L than in those below 15 mg/L (OR 10.24, p = 0.048). Conclusions: In critically ill ICU patients, calculator-derived AUC_0–24/MIC showed significant discordance with single-trough vancomycin monitoring, primarily indicating apparent under-exposure by trough criteria in patients with adequate AUC_0–24/MIC levels, which may lead to unnecessary dose escalation. Full article

(This article belongs to the Special Issue Innovative Tools for Drug Analysis and Therapeutic Drug Monitoring (TDM): 2nd Edition)

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21 pages, 16473 KB

Open AccessArticle

In Silico Docking and Spectroscopic Evaluation of a Thiocarbohydrazone Derivative: Structural Elucidation and Enzyme Inhibitory Mechanisms

by Maria Karatzia, Nikitas Georgiou, Ektoras Vasileios Apostolou, Eleftherios Papamichalis, Sophia C. Hayes, Thomas Mavromoustakos and Demeter Tzeli

Pharmaceuticals 2026, 19(7), 1108; https://doi.org/10.3390/ph19071108 - 17 Jul 2026

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Abstract

Objectives: Thiocarbohydrazones represent an important class of Schiff base derivatives with versatile chemical and biological properties. Methods: Herein, we present a combined in silico spectroscopic and molecular docking investigation of N′-benzylidenehydrazinecarbothiohydrazide (1). Results: Conformational docking studies were conducted against cathepsin B, acetylcholinesterase, HER2, [...] Read more.

Objectives: Thiocarbohydrazones represent an important class of Schiff base derivatives with versatile chemical and biological properties. Methods: Herein, we present a combined in silico spectroscopic and molecular docking investigation of N′-benzylidenehydrazinecarbothiohydrazide (1). Results: Conformational docking studies were conducted against cathepsin B, acetylcholinesterase, HER2, protein kinase C, and protein kinase A. The compound displayed favorable binding affinities and key interactions within the catalytic sites of all targets, with the strongest predicted binding observed for acetylcholinesterase. Notably, all conformers exhibited higher affinity for protein kinase C than the reference inhibitor balanol, and hydroxylation led to an approximately 10% enhancement in docking performance. Density functional theory (DFT) calculations were employed to analyze vibrational properties, and IR and Raman spectra were computed to elucidate structural features and conformational behavior. Conclusions: The integrated spectroscopic and docking analyses provide mechanistic insights into ligand–target interactions and support rational drug design. These findings identify thiocarbohydrazone derivatives as promising multi-target candidates for the development of enzyme inhibitors relevant to neurodegenerative, oncological, and inflammatory diseases. Full article

(This article belongs to the Special Issue Active Molecule Design Based on Conformational Pharmacology: Synthesis, NMR Spectroscopy, and Computational Modeling)

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30 pages, 34687 KB

Open AccessArticle

Material Basis and Mechanisms of Action of PuRenDan in the Treatment of Type 2 Diabetes Mellitus: An Integrated Network Pharmacology and Molecular Simulation Study

by Wenshuai Yang, Gaojie Ouyang, Wenwen Zhou, Binan Lu and Zongran Pang

Pharmaceuticals 2026, 19(7), 1107; https://doi.org/10.3390/ph19071107 - 17 Jul 2026

Viewed by 374

Abstract

Background/Objectives: Type 2 diabetes mellitus (T2DM) is a chronic multifactorial metabolic disorder requiring multi-target therapeutic strategies. This study aimed to predict the potential material basis, key targets and molecular mechanisms by which PuRenDan (PRD) may act against T2DM through an integrated network [...] Read more.

Background/Objectives: Type 2 diabetes mellitus (T2DM) is a chronic multifactorial metabolic disorder requiring multi-target therapeutic strategies. This study aimed to predict the potential material basis, key targets and molecular mechanisms by which PuRenDan (PRD) may act against T2DM through an integrated network pharmacology and molecular simulation approach. Methods: Active compounds of PRD were screened from TCMSP, HERB 2.0 and the literature, and compound-related targets were predicted using TCMSP, SwissTargetPrediction and PharmMapper. T2DM-associated targets were collected from OMIM, DrugBank, DisGeNET, HPO, ClinPGx and GeneCards to obtain drug–disease intersection targets. Cytoscape was used to construct herb–compound–target and protein–protein interaction (PPI) networks, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Molecular docking was performed using AutoDock Vina1.1.2, and representative ligand–receptor complexes were further assessed by 100 ns molecular dynamics (MD) simulations and molecular mechanics/Poisson–Boltzmann surface area (MM/PBSA) binding free-energy analysis. Results: A total of 163 active compounds, 597 PRD-related targets, 9138 T2DM-associated targets and 483 intersection targets were identified. β-sitosterol, emodin, quercetin, kaempferol and formononetin were predicted as major active compounds, whereas AKT1, TP53, SRC, IL6, TNF, EGFR and ESR1 were identified as disease-related network hubs. KEGG enrichment highlighted the PI3K-Akt, MAPK, HIF-1, FoxO, mTOR, AGE-RAGE and TNF signalling pathways. Docking predicted a comparatively favourable multi-target binding tendency for β-sitosterol. MD and MM/PBSA analyses further suggested favourable dynamic stability for β-sitosterol-TNF, β-sitosterol-AKT1, β-sitosterol-SRC and emodin-EGFR complexes, with β-sitosterol-TNF showing the lowest predicted binding free energy among the simulated systems. Conclusions: These in silico findings suggest that PRD may regulate T2DM-related inflammatory, insulin-signalling, oxidative-stress and metabolic networks through coordinated multi-compound, multi-target and multi-pathway actions. β-sitosterol may represent an important candidate material basis of PRD, with TNF, AKT1, SRC and EGFR as potential key targets. These conclusions remain predictive and require validation in biochemical, cellular and animal experiments. Full article

(This article belongs to the Topic Biological and Pharmacological Activity of Natural Products)

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21 pages, 4299 KB

Open AccessArticle

Functional Characterization and Inhibition Analysis of a Glutathione Transferase from Cryptosporidium parvum: A Potential Target for Antiparasitic Drug Development

by Panagiota D. Pantiora, Nikolaos D. Georgakis, Dimitris Matiadis, Marina Sagnou and Nikolaos E. Labrou

Pharmaceuticals 2026, 19(7), 1106; https://doi.org/10.3390/ph19071106 - 17 Jul 2026

Viewed by 314

Abstract

Background/Objectives: Cryptosporidiosis, caused by Cryptosporidium parvum, is a significant cause of diarrheal disease, particularly affecting young children and immunocompromised individuals. With current treatments offering limited efficacy, there is an urgent need for novel therapeutic targets. Methods: In this study, we [...] Read more.

Background/Objectives: Cryptosporidiosis, caused by Cryptosporidium parvum, is a significant cause of diarrheal disease, particularly affecting young children and immunocompromised individuals. With current treatments offering limited efficacy, there is an urgent need for novel therapeutic targets. Methods: In this study, we report the cloning, expression, and functional characterization of a glutathione transferase (GST) from C. parvum (CpGST). Results: Biocomputing analysis revealed a single gene encoding a cytosolic enzyme with distinct structural features, compared to human cytosolic homologs. Structural modeling indicated a non-canonical thioredoxin fold and a truncated C-terminal domain, suggesting functional divergence. CpGST was expressed in Escherichia coli, and its enzymatic properties were characterized. Although the enzyme displayed a narrow substrate spectrum, it showed a distinct substrate preference, retaining catalytic activity toward the standard GST substrates 1-chloro-2,4-dinitrobenzene (CDNB) and cumene hydroperoxide (CuOOH). Steady-state kinetic analysis revealed limited affinity for both reduced glutathione (GSH) and CDNB. Inhibition analysis identified several polyphenols and synthetic curcumin analogues as potent inhibitors, with IC₅₀ values in the low micromolar range. Kinetic analysis with the most potent inhibitor revealed a mixed-type inhibition mechanism. Conclusions: These findings support the classification of CpGST as a structurally and functionally distinct member of the GST family, likely adapted to the parasite’s physiology and metabolism. The enzyme’s divergence from human GSTs, along with its favorable druggability profile, underscores its potential as a target for anti-cryptosporidial drug development, particularly in strategies aimed at disrupting stress response and detoxification pathways. Full article

(This article belongs to the Special Issue Advances in the Mechanism of Action and the Therapeutic Role of Phytopharmaceuticals)

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17 pages, 2167 KB

Open AccessArticle

Development and Evaluation of Physiologically Based Pharmacokinetic (PBPK) Models to Investigate the Effect of CYP2D6 Polymorphism on Metoclopramide Systemic Exposure

by Iqra Shahzad, Ammara Zamir, Muhammad Fawad Rasool, Amer S. Alali, Iltaf Hussain and Faleh Alqahtani

Pharmaceuticals 2026, 19(7), 1105; https://doi.org/10.3390/ph19071105 - 17 Jul 2026

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Abstract

Background: Physiologically based pharmacokinetic (PBPK) modeling is a mechanistic tool used to predict how a drug moves through the body by incorporating real human physiology, including organ sizes, blood flows, tissue compositions, and enzyme activities. It has been widely employed to estimate drug [...] Read more.

Background: Physiologically based pharmacokinetic (PBPK) modeling is a mechanistic tool used to predict how a drug moves through the body by incorporating real human physiology, including organ sizes, blood flows, tissue compositions, and enzyme activities. It has been widely employed to estimate drug exposure in different populations with organ impairment, genotype variabilities, and physiological variations. Metoclopramide is an antiemetic and prokinetic agent that is subject to CYP2D6 polymorphism. The study aims to develop PBPK models for several CYP2D6 variants to predict changes in the pharmacokinetic (PK) behavior of metoclopramide. Methods: To conduct this study, a literature review was conducted, and the retrieved physicochemical, biochemical, and PK data were integrated into PK-Sim to develop a PBPK model. Initially, a non-genotype-specific model was developed and extrapolated to genotype-based models. The models were verified using a Visual Predicted Check (VPC), mean predicted-to-observed ratio (R_pre/obs) values, and mean relative deviation (MRD). Results: The simulated profiles were aligned with the reported data, and all the predicted and observed PK parameters were comparable, as the R_pre/obs values were within the 0.5–2 range and MRD values were <2. Moreover, an increasing trend in AUC_0–∞ was observed across CYP2D6*wt/*wt, CYP2D6*wt/*10, CYP2D6*10/*10, and CYP2D6*5/*10, with approximately 1.63-, 2.64-, and 2.88-fold increases compared with the CYP2D6*wt/*wt genotype. Conclusions: The models have adequately estimated the PK behavior of metoclopramide across different CYP2D6 variants. These models might be helpful for populations with diverse CYP2D6 genotypes in dose optimization. Full article

(This article belongs to the Special Issue Population Pharmacokinetics and Pharmacogenetics, 2nd Edition)

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17 pages, 5008 KB

Open AccessReview

Deconstructing the Master Switch: Advances in Direct NLRP3 Inhibition

by Yiming Xu and Sasha Murphy

Pharmaceuticals 2026, 19(7), 1104; https://doi.org/10.3390/ph19071104 - 17 Jul 2026

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Abstract

As a bona fide “master switch,” NOD-like receptor family pyrin domain containing 3 (NLRP3) functions not only as an inflammatory mediator but also as a primary sensor of metabolic stress and danger signals. Its dysregulation has been implicated in an exceptionally broad spectrum [...] Read more.

As a bona fide “master switch,” NOD-like receptor family pyrin domain containing 3 (NLRP3) functions not only as an inflammatory mediator but also as a primary sensor of metabolic stress and danger signals. Its dysregulation has been implicated in an exceptionally broad spectrum of human diseases, making it one of the most intensively studied therapeutic targets. While the discovery of the first direct antagonist MCC950 marked a turning point, the subsequent explosion of diverse inhibitor classes demands a systematic, up-to-date evaluation. This review comprehensively analyzes the current landscape of direct NLRP3 inhibitors, focusing on how recent structural breakthroughs illuminate specific mechanism-of-action differences. We systematically reviewed peer-reviewed literature from 2015 to 2026, categorizing small-molecule inhibitors based on their chemical scaffolds and binding pockets as revealed by cryo-EM and X-ray crystallography data. By mapping these structural insights into functional outcomes, we provide a definitive molecular-level analysis designed to guide the rational design and optimization of next-generation NLRP3-targeted therapeutics. Full article

(This article belongs to the Special Issue Targeting the NLRP3: From Novel Inhibitor Design to Clinic Trials and Therapeutic Applications)

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31 pages, 5454 KB

Open AccessArticle

Development and Characterization of Honey- and Essential Oil-Based Structured Systems for Skin Applications

by Corina-Bianca Ioniță-Mîndrican, Manuela Ghica, Ancuța Cătălina Fița, Eliza Oprea, Mihaela Buleandră, Irinel Adriana Badea, Cristina-Ionela Stănciulescu, Emma Adriana Ozon, Silviu-Iulian Filipiuc and Carolina Negrei

Pharmaceuticals 2026, 19(7), 1103; https://doi.org/10.3390/ph19071103 - 17 Jul 2026

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Abstract

Background: The present study aimed to develop and characterize three honey- and essential oil-based structured systems intended for topical skin application. Materials and Methods: The semisolid systems were prepared as oil-in-water structured emulsions containing four types of honey (Manuka, Tualang, chestnut, [...] Read more.

Background: The present study aimed to develop and characterize three honey- and essential oil-based structured systems intended for topical skin application. Materials and Methods: The semisolid systems were prepared as oil-in-water structured emulsions containing four types of honey (Manuka, Tualang, chestnut, and manna), three essential oils (palmarosa, cistus, and lavender), and five vegetable oils (pomegranate seed, aloe, centella, hemp seed, and calendula). Each formulation consisted of two honey types, one essential oil, and two vegetable oils, with final concentrations of 5% honey and 0.1–0.2% essential oil. The formulations were investigated through physicochemical, rheological, and in vivo skin evaluations. Results: Rheological analysis demonstrated non-Newtonian pseudoplastic behavior with shear-thinning and thixotropic characteristics, indicating that the systems are structured, semisolid, and suitable for topical application. Differences in spreadability and consistency suggested variations in the internal organization of the emulsion matrices. In vivo skin assessments were performed over four weeks using non-invasive instrumental methods. The obtained results demonstrated improvements in skin hydration, elasticity, firmness, and skin barrier function, together with reductions in transepidermal water loss and erythema. The evaluation of skin textural parameters revealed improvements in skin uniformity and a reduction in the appearance of wrinkles. Among the tested structured systems, F3 formulation exhibited the most pronounced moisturizing effect, while F1 formulation showed notable improvements in parameters associated with skin texture and wrinkle-related features. Conclusions: Overall, the results indicate that honey-based topical systems enriched with essential and vegetable oils represent promising multifunctional semisolid formulations for topical skin-conditioning and barrier-supportive applications. Their favorable rheological behavior, combined with beneficial effects on skin hydration, barrier function, and skin surface properties, supports their potential use in skin-conditioning and anti-aging-related formulations. Full article

(This article belongs to the Special Issue Natural Products for Skin Applications)

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28 pages, 11962 KB

Open AccessArticle

Bioactive Silver Nanoparticles Synthesized Using Endophytic Bacillus subtilis CG1 and Their Antimicrobial and Antibiofilm Potential Against Drug-Resistant Pathogens

by Ghaida Saud Aljohani and Saleh H. Salmen

Pharmaceuticals 2026, 19(7), 1102; https://doi.org/10.3390/ph19071102 - 17 Jul 2026

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Abstract

Background/Objectives: The study addresses the global health challenge posed by multidrug-resistant (MDR) pathogens, highlighting the urgent need for alternative antimicrobial solutions. This study investigated the in vitro antimicrobial and antibiofilm potential of endophytic mediated-synthesized silver nanoparticles (AgNPs). Methods: An endophytic bacterium was isolated [...] Read more.

Background/Objectives: The study addresses the global health challenge posed by multidrug-resistant (MDR) pathogens, highlighting the urgent need for alternative antimicrobial solutions. This study investigated the in vitro antimicrobial and antibiofilm potential of endophytic mediated-synthesized silver nanoparticles (AgNPs). Methods: An endophytic bacterium was isolated from the medicinal plant Commiphora gileadensis in Saudi Arabia and identified as Bacillus subtilis CG1 through 16S rRNA gene sequencing. The bacterium was utilized for the green synthesis of AgNPs, as confirmed by Ultraviolet-visible (UV–Vis) spectroscopy. AgNPs characterization was done using Fourier-transform infrared (FTIR) spectroscopy, Transmission and scanning electron microscopy (TEM and SEM), energy-dispersive X-ray spectroscopy (EDX), and dynamic light scattering (DLS). The antimicrobial efficacy of the fabricated AgNPs was tested against eight clinically relevant pathogens using standard in vitro assays such as the agar disk diffusion method, minimum inhibitory concentration (MIC), minimum bactericidal and fungicidal concentrations (MBC and MFC). Additionally, AgNPs were tested for antibiofilm activity against P. aeruginosa and S. epidermidis. Tested pathogens included Methicillin-Resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Candida auris, Candida albicans, and Candida tropicalis. The antibiofilm efficacy was tested using the Crystal violet assay. Results: UV–Vis spectroscopy confirmed AgNP formation with a characteristic absorption peak at 412 nm. FTIR analysis identified the presence of hydroxyl, nitrile, and alkyne functional groups, which are involved in nanoparticle reduction and stabilization. TEM and SEM revealed predominantly spherical AgNPs with sizes ranging from 17 to 72 nm, while EDX confirmed silver as the major elemental component. DLS analysis showed a Z-average particle size of 113.9 ± 67.75 nm and a zeta potential of −24.2 mV. The synthesized AgNPs exhibited concentration-dependent antimicrobial activity, producing inhibition zones of 10–20 mm at 240 µg/mL. MIC values ranged from 6.25 to 25 µg/mL, whereas MBC and MFC values ranged from 6.25 to 50 µg/mL and 25 to 100 µg/mL, respectively. Moreover, bacterial growth kinetics analysis demonstrated a concentration-dependent inhibition of growth by AgNPs at MIC and sub-MIC concentrations. Additionally, AgNPs demonstrated significant antibiofilm activity against P. aeruginosa and S. epidermidis.Conclusions: Overall, B. subtilis CG1-mediated AgNPs exhibited promising physicochemical properties and antimicrobial and antibiofilm activities, suggesting their potential as alternatives for combating resistant and biofilm-associated infections. Full article

(This article belongs to the Section Medicinal Chemistry)

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Pharmaceuticals, Volume 19, Issue 7 (July 2026) – 159 articles

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