Pharmaceuticals doi: 10.3390/ph17121673

Authors: Nicholas Geremia Federico Giovagnorio Agnese Colpani Andrea De Vito Alexandru Botan Giacomo Stroffolini Dan-Alexandru Toc Verena Zerbato Luigi Principe Giordano Madeddu Roberto Luzzati Saverio Giuseppe Parisi Stefano Di Bella

Background: Biofilm-associated infections frequently span multiple body sites and represent a significant clinical challenge, often requiring a multidisciplinary approach involving surgery and antimicrobial therapy. These infections are commonly healthcare-associated and frequently related to internal or external medical devices. The formation of biofilms complicates treatment, as they create environments that are difficult for most antimicrobial agents to penetrate. Fluoroquinolones play a critical role in the eradication of biofilm-related infections. Numerous studies have investigated the synergistic potential of combining fluoroquinolones with other chemical agents to augment their efficacy while minimizing potential toxicity. Comparative research suggests that the antibiofilm activity of fluoroquinolones is superior to that of beta-lactams and glycopeptides. However, their activity remains less effective than that of minocycline and fosfomycin. Noteworthy combinations include fluoroquinolones with fosfomycin and aminoglycosides for enhanced activity against Gram-negative organisms and fluoroquinolones with minocycline and rifampin for more effective treatment of Gram-positive infections. Despite the limitations of fluoroquinolones due to the intrinsic characteristics of this antibiotic, they remain fundamental in this setting thanks to their bioavailability and synergisms with other drugs. Methods: A comprehensive literature search was conducted using online databases (PubMed/MEDLINE/Google Scholar) and books written by experts in microbiology and infectious diseases to identify relevant studies on fluoroquinolones and biofilm. Results: This review critically assesses the role of fluoroquinolones in managing biofilm-associated infections in various clinical settings while also exploring the potential benefits of combination therapy with these antibiotics. Conclusions: The literature predominantly consists of in vitro studies, with limited in vivo investigations. Although real world data are scarce, they are in accordance with fluoroquinolones’ effectiveness in managing early biofilm-associated infections. Also, future perspectives of newer treatment options to be placed alongside fluoroquinolones are discussed. This review underscores the role of fluoroquinolones in the setting of biofilm-associated infections, providing a comprehensive guide for physicians regarding the best use of this class of antibiotics while highlighting the existing critical issues.

Pharmaceuticals doi: 10.3390/ph17121672

Authors: Ruvimbo Faith Tapera Xavier Siwe-Noundou Leshweni Jeremia Shai Shoeshoe Mokhele

Helianthus tuberosus L. (Jerusalem artichoke) tubers and aerial parts possess both nutritional and therapeutic properties. The Jerusalem artichoke has been utilized for various applications, including its use as a functional food source, a reservoir of bioactive compounds, and a raw material to produce biofuels. Moreover, the Jerusalem artichoke is a rich source of an indigestible polysaccharide called inulin, which serves as a prebiotic that improves gastrointestinal health. This plant has been used globally throughout history as a dietary supplement, for pain treatment, to reduce swelling, and to boost the immune system, as well as to treat skin wounds in folk medicine. It is an abundant source of bioactive compounds, such as phenolic acids, coumarins, and flavonoids, which are known to exert pharmacological activities, including antioxidant, antimicrobial, and anti-inflammatory properties. The literature on its potential as an antidiabetic, anticancer, anti-fungistatic, antiviral, and anti-obesity agent, among others, is scanty. This review aims to provide a comprehensive overview of Helianthus tuberosus L.’s traditional uses, nutritional properties, secondary bioactive compounds, and pharmacological properties to further explore its health benefits.

Pharmaceuticals doi: 10.3390/ph17121671

Authors: Renata Torres Pessoa Lucas Yure Santos da Silva Isabel Sousa Alcântara Tarcísio Mendes Silva Eduardo dos Santos Silva Roger Henrique Sousa da Costa Aparecida Barros da Silva Jaime Ribeiro-Filho Anita Oliveira Brito Pereira Bezerra Martins Henrique Douglas Melo Coutinho Jean Carlos Pereira Sousa Andréa Rodrigues Chaves Ricardo Neves Marreto Irwin Rose Alencar de Menezes

Background/Objectives: This study evaluated the antinociceptive effect of the Ximenia americana L. bark extract (HEXA) and its primary component, caffeic acid (CA), through in vivo assays. Methods: The antinociceptive properties were assessed using abdominal writhing, hot plate, and Von Frey tests. Additionally, the study investigated the modulation of various pain signaling pathways using a pharmacological approach. Results: The results demonstrated that all doses of the HEXA significantly increased latency in the hot plate test, decreased the number of abdominal contortions, reduced hyperalgesia in the Von Frey test, and reduced both phases of the formalin test. Caffeic acid reduced licking time in the first phase of the formalin test at all doses, with the highest dose showing significant effects in the second phase. The HEXA potentially modulated α2-adrenergic (52.99%), nitric oxide (57.77%), glutamatergic (33.66%), vanilloid (39.84%), cyclic guanosine monophosphate (56.11%), and K+ATP channel-dependent pathways (38.70%). Conversely, CA influenced the opioid, glutamatergic (53.60%), and vanilloid (34.42%) pathways while inhibiting nitric oxide (52.99%) and cyclic guanosine monophosphate (38.98%). Conclusions: HEXA and CA exhibit significant antinociceptive effects due to their potential interference in multiple pain signaling pathways. While the molecular _targets remain to be fully investigated, HEXA and CA demonstrate significant potential for the development of new analgesic drugs.

Pharmaceuticals doi: 10.3390/ph17121669

Authors: Mateen A. Khan

The hallmark of Alzheimer’s disease (AD) is the buildup of amyloid-β (Aβ), which is produced when the amyloid precursor protein (APP) misfolds and deposits as neurotoxic plaques in the brain. A functional iron responsive element (IRE) RNA stem loop is encoded by the APP 5′-UTR and may be a _target for regulating the production of Alzheimer’s amyloid precursor protein. Since modifying Aβ protein expression can give anti-amyloid efficacy and protective brain iron balance, _targeted regulation of amyloid protein synthesis through modulation of 5′-UTR sequence function is a novel method for the prospective therapy of Alzheimer’s disease. Numerous mRNA interference strategies _target the 2D RNA structure, even though messenger RNAs like tRNAs and rRNAs can fold into complex, three-dimensional structures, adding even another level of complexity. The IRE family is among the few known 3D mRNA regulatory elements. This review seeks to describe the structural and functional aspects of IREs in transcripts, including that of the amyloid precursor protein, that are relevant to neurodegenerative diseases, including AD. The mRNAs encoding the proteins involved in iron metabolism are controlled by this family of similar base sequences. Like ferritin IRE RNA in their 5′-UTR, iron controls the production of APP in their 5′-UTR. Iron misregulation by iron regulatory proteins (IRPs) can also be investigated and contrasted using measurements of the expression levels of tau production, Aβ, and APP. The development of AD is aided by iron binding to Aβ, which promotes Aβ aggregation. The development of small chemical therapeutics to control IRE-modulated expression of APP is increasingly thought to _target messenger RNAs. Thus, IRE-modulated APP expression in AD has important therapeutic implications by _targeting mRNA structures.

Pharmaceuticals doi: 10.3390/ph17121668

Authors: Michele Tonelli Anna Sparatore Ivan Bassanini Valeria Francesconi Fabio Sparatore Kevin K. Maina Serena Delbue Sarah D’Alessandro Silvia Parapini Nicoletta Basilico

Background/Objectives: Four years after the COVID-19 pandemic, a very limited number of drugs has been marketed; thus, the search for new medications still represents a compelling need. In our previous work on antiviral, antiparasitic, and antiproliferative agents, we described several compounds (1–13 and 16–20) structurally related to clofazimine, chloroquine, and benzimidazole derivatives. Thus, we deemed it worthwhile to test them against the replication of SARS-CoV-2, together with a few other compounds (14, 15 and 21–25), which showed some analogy to miscellaneous anti-coronavirus agents. Methods: Twenty-five structurally assorted compounds were evaluated in vitro for cytotoxicity against Vero E6 and for their ability to inhibit SARS-CoV-2 replication. Results: Several compounds (2, 3, 10, 11, 13–15, 18–20) demonstrated antiviral activity (IC50 range 1.5–28 µM) and six of them exhibited an interesting selectivity index in the range 4.5–20. The chloroquine analogs 10 and 11 were more potent than the reference chloroquine itself and doubled its SI value (20 versus 11). Also, the benzimidazole ring emerged as a valuable scaffold, originating several compounds (13–15 and 18–20) endowed with anti-SARS-CoV-2 activity. Despite the modest activity, the cytisine and the arylamino enone derivatives 23 and 25, respectively, also deserve further consideration as model compounds. Conclusions: The investigated chemotypes may represent valuable hit compounds, deserving further in-depth biological studies to define their mechanisms of action. The derived information will guide the subsequent chemical optimization towards the development of more efficient anti-SARS-CoV-2 agents.

Pharmaceuticals doi: 10.3390/ph17121667

Authors: Mohamed H. Attia Deena S. Lasheen Nermin Samir Azza T. Taher Hatem A. Abdel-Aziz Dalal A. Abou El Ella

Background: The increasing prevalence of drug resistance in cancer therapy underscores the urgent need for novel therapeutic approaches. Dual enzyme inhibitors, _targeting critical kinases such as CDK2 and TRKA, represent a promising strategy. The goal of this investigation was to design, synthesize, and evaluate a set of pyrazolo[1,5-a]pyrimidine derivatives for their dual inhibition potential toward CDK2 and TRKA kinases, along with their potential antiproliferative against cancer cell lines. Methods: A set of pyrazolo[1,5-a]pyrimidine derivatives (6a–t, 11a–g, and 12) was synthesized and subjected to in vitro enzymatic assays to determine their inhibitory activity against CDK2 and TRKA kinases. Selected compounds were further assessed for antiproliferative effects across the set of 60 cell lines from the NCI, representing various human cancer types. Additionally, simulations of molecular docking were conducted to explore the modes of binding for the whole active compounds and compare them with known inhibitors. Results: Compounds 6t and 6s exhibited potent dual inhibitory activity, showing an IC50 = 0.09 µM and 0.23 µM against CDK2, and 0.45 µM against TRKA, respectively. These results were comparable to reference inhibitors ribociclib (CDK2, IC50 = 0.07 µM) and larotrectinib (TRKA, IC50 = 0.07 µM). Among the studied derivatives, compound 6n displayed a notable broad-spectrum anticancer activity, achieving a mean growth inhibition (GI%) of 43.9% across 56 cell lines. Molecular docking simulations revealed that the synthesized compounds adopt modes of binding similar to those of the lead inhibitors. Conclusions: In this study, prepared pyrazolo[1,5-a]pyrimidine derivatives demonstrated significant potential as dual CDK2/TRKA inhibitors, and showed potent anticancer activity toward diverse cancer cell lines. These findings highlight their potential as key compounds for the design of novel anticancer therapeutics.

Pharmaceuticals doi: 10.3390/ph17121666

Authors: Jian Wang Nianhui Yu Yunpeng Tang Yingsheng Cheng Hui Li

Background: Albumin-bound paclitaxel (nab-PTX) nanoparticles have been proven effective in treating advanced pancreatic cancer. However, the clinical application of nab-PTX nanoparticles is often associated with suboptimal outcomes and severe side effects due to its non-specific distribution and rapid clearance. This study aims to develop a novel nanoplatform that integrates sonodynamic therapy (SDT) and chemotherapy to enhance treatment efficacy and reduce systemic side effects. Methods: Bovine serum albumin (BSA) was conjugated with chlorin e6 and paclitaxel (PTX) to form stable nanoparticles (NPs). These NPs were then incorporated into a biodegradable poly(lactic-co-glycolic acid)–b-polyethylene glycol–b–poly(lactic-co-glycolic acid) hydrogel for _targeted drug delivery. The system’s stability and drug release profile were analyzed, followed by in vitro studies to evaluate cellular uptake and cancer cell killing efficacy. In vivo evaluation was performed using pancreatic cancer xenograft models, with intratumoral injection of the drug-loaded hydrogel. Results: The developed hydrogel system demonstrated enhanced stability and sustained release of PTX. In vitro analyses revealed significant cellular uptake and synergistic cancer cell killing effects through combined SDT and chemotherapy. In vivo studies showed prolonged intratumoral retention of the drug and remarkable inhibition of tumor growth. Conclusions: This novel nanoplatform offers a promising approach for improving pancreatic cancer treatment by enhancing intratumoral drug retention and minimizing systemic side effects. The synergistic effects of SDT and chemotherapy demonstrate the potential of this strategy in achieving better therapeutic outcomes.

Pharmaceuticals doi: 10.3390/ph17121665

Authors: Yuqi Fu Le Yang Lei Liu Ling Kong Hui Sun Ye Sun Fengting Yin Guangli Yan Xijun Wang

Rhein is a natural active ingredient in traditional Chinese medicine that has attracted much attention due to its wide range of pharmacological activities. However, its clinical application is limited by low water solubility, poor oral absorption, and potential toxicity to the liver and kidneys. Recently, advanced extraction and synthesis techniques have made it possible to develop derivatives of rhein, which have better pharmacological properties and lower toxicity. This article comprehensively summarizes the biological activity and action mechanism of rhein. Notably, we found that TGF-β1 is the _target of rhein improving tissue fibrosis, while NF-κB is the main _target of its anti-inflammatory effect. Additionally, we reviewed the current research status of the pharmacokinetics, toxicology, structural optimization, and potential drug applications of rhein and found that the coupling and combination therapy of rhein and other active ingredients exhibit a synergistic effect, significantly enhancing therapeutic efficacy. Finally, we emphasize the necessity of further studying rhein’s pharmacological mechanisms, toxicology, and development of analogs, aiming to lay the foundation for its widespread clinical application as a natural product and elucidate its prospects in modern medicine.

Pharmaceuticals doi: 10.3390/ph17121664

Authors: Somaia S. Abd El-Karim Manal M. Anwar Yasmin M. Syam Hassan M. Awad Asmaa Negm El-Dein Mohamed K. El-Ashrey Hamad M. Alkahtani Sameh H. Abdelwahed

Background/Objectives: The alarming rise in antibiotic resistance necessitates the discovery of novel antimicrobial agents. This study aims to design, synthesize, and evaluate new benzofuran–pyrazole-based compounds for their antimicrobial, antioxidant, and anti-inflammatory properties. Methods: New benzofuran–pyrazole hybrid molecules were synthesized using the Vilsmeier–Haach reaction and other chemical processes. The structures of the synthesized compounds were confirmed through micro-analytical and spectral analyses. Their antimicrobial activities were assessed against various bacterial and fungal strains, while antioxidant and anti-inflammatory properties were evaluated using DPPH-free radical scavenging and HRBC membrane stabilization assays, respectively. The most promising compounds were further tested for DNA gyrase B inhibition. Results: Compounds 9, 10, and 11b–d exhibited significant broad-spectrum antimicrobial activity with MIC values ranging from 2.50 to 20 µg/mL. Compounds 4, 6, 9, 11b, and 11d demonstrated high antioxidant activity, with DPPH scavenging percentages between 84.16% and 90.52%. Most compounds showed substantial anti-inflammatory effects, with HRBC membrane stabilization percentages ranging from 86.70% to 99.25%. Compound 9 notably inhibited E. coli DNA gyrase B with an IC50 of 9.80 µM, comparable to ciprofloxacin. Conclusions: The benzofuran–pyrazole-based compounds, particularly compound 9, show great potential as new antimicrobial agents due to their broad-spectrum activity and potent DNA gyrase B inhibition. These findings support further development and optimization of these compounds for clinical applications.

Pharmaceuticals doi: 10.3390/ph17121663

Authors: Hui-Wen Chan Deng-Yu Kuo Pei-Wei Shueng Hui-Yen Chuang

The tumor microenvironment (TME) is a critical factor in cancer progression, driving tumor growth, immune evasion, therapeutic resistance, and metastasis. Understanding the dynamic interactions within the TME is essential for advancing cancer management. Molecular imaging provides a non-invasive, real-time, and longitudinal approach to studying the TME, with techniques such as positron emission tomography (PET), magnetic resonance imaging (MRI), and fluorescence imaging offering complementary strengths, including high sensitivity, spatial resolution, and intraoperative precision. Recent advances in imaging probe development have enhanced the ability to _target and monitor specific components of the TME, facilitating early cancer diagnosis, therapeutic monitoring, and deeper insights into tumor biology. By integrating these innovations, molecular imaging offers transformative potential for precision oncology, improving diagnostic accuracy and treatment outcomes through a comprehensive assessment of TME dynamics.

Pharmaceuticals doi: 10.3390/ph17121661

Authors: Da Hyeon An Chan Ho Lee Yeeun Kwon Tae Hee Kim Eun Ji Kim Jae In Jung Sangil Min Eun Ju Cheong Sohyun Kim Hee Kyu Kim Sun Eun Choi

Background/Objectives: Sarcopenia is characterized by the loss of muscle mass and function, increases in mortality rate, and risk of comorbidities in the elderly. This study evaluated the effects of Alnus japonica hot water extract (AJHW) and its active compound, oregonin, on muscle atrophy and apoptosis in vitro. Methods: AJHW underwent phytochemical analysis. C2C12 cells were subjected to H2O2 and dexamethasone to induce oxidative stress and muscle loss, after which AJHW and oregonin were administered to assess their impacts on cell viability, apoptosis, muscle protein synthesis stimulation, and muscle protein degradation inhibition. Cell viability was assessed via an MTT assay, and apoptosis was analyzed by measuring Bcl-2, Bax, cleaved caspase-3, and cleaved PARP through Western blotting. Western blotting and RT-PCR were utilized to analyze MyoD, Myogenin, Atrogin-1, and MuRF1 protein and gene expression in a muscle atrophy model, as well as the Akt/mTOR and FoxO3α pathways. Results: AJHW was confirmed to contain oregonin, an active compound. AJHW and oregonin significantly increased cell viability and reduced apoptosis by upregulating Bcl-2 and downregulating Bax, cleaved caspase-3, and cleaved PARP. They significantly enhanced muscle protein synthesis through the upregulation of MyoD and Myogenin, while diminishing muscle degradation by downregulating Atrogin-1 and MuRF1. The activation of the Akt/mTOR pathway and inhibition of the FoxO3α pathway were also observed. Conclusions: AJHW and oregonin effectively prevented muscle cell apoptosis, promoted muscle protein synthesis, and inhibited muscle protein degradation in vitro. These results suggest that AJHW and oregonin could serve as therapeutic agents to prevent and treat sarcopenia.

Pharmaceuticals doi: 10.3390/ph17121662

Authors: Karen Geoffroy Mélissa Viens Emma Mary Kalin Zied Boudhraa Dominic Guy Roy Jian Hui Wu Diane Provencher Anne-Marie Mes-Masson Marie-Claude Bourgeois-Daigneault

Background/Objectives: Ovarian cancer is the deadliest gynecologic cancer, and with the majority of patients dying within the first five years of diagnosis, new therapeutic options are required. The small guanosine triphosphatase (GTPase) Ras-related nuclear protein (Ran) has been reported to be highly expressed in high-grade serous ovarian cancers (HGSOCs) and associated with poor outcomes. Blocking Ran function or preventing its expression were shown to be promising treatment strategies, however, there are currently no small molecule inhibitors available to specifically inhibit Ran function. Interestingly, a previous study suggested that the Vesicular stomatitis virus (VSV) could inhibit Ran activity. Given that VSV is an oncolytic virus (OV) and, therefore, has anti-cancer activity, we reasoned that oncolytic VSV (oVSV) might be particularly effective against ovarian cancer via Ran inhibition. Methods: We evaluated the sensitivity of patient-derived ovarian cancer cell lines to oVSV, as well as the impact of oVSV on Ran and vice versa, using overexpression systems, small interfering RNAs (siRNAs), and drug inhibition. Results: In this study, we evaluated the interplay between oVSV and Ran and found that, although oVSV does not consistently block Ran, increased Ran activation allows for better oVSV replication and tumor cell killing. Conclusions: Our study reveals a positive impact of Ran on oVSV sensitivity. Given the high expression of Ran in HGSOCs, which are particularly aggressive ovarian cancers, our data suggest that oVSV could be effective against the deadliest form of the disease.

Pharmaceuticals doi: 10.3390/ph17121660

Authors: Neetu Dayal Riddhi Chaudhuri Kofi Simpa Yeboah Nickolas R. Brauer Herman O. Sintim

Background: The protein kinases CLK and ROCK play key roles in cell growth and migration, respectively, and are potential anticancer _targets. ROCK inhibitors have been approved by the FDA for various diseases and CLK inhibitors are currently being trialed in the clinic as anticancer agents. Compounds with polypharmacology are desired, especially in oncology, due to the potential for high efficacy as well as addressing resistance issues. In this report, we have identified and characterized novel, boron-containing dual CLK/ROCK inhibitors with promising anticancer properties. Methods: A library of boronic acid-based CLK/ROCKi was synthesized via Povarov/Doebner-type multicomponent reactions. Kinase inhibition screening and cancer cell viability assays were performed to identify the hit compounds. To gain insights into the probable binding modes of the compounds to the kinases, docking studies were performed. Cell cycle analysis, qPCR and immunoblotting were carried out to further characterize the mode(s) of action of the lead candidates. Results: At 25 nM, the top compounds HSD1400 and HSD1791 inhibited CLK1 and 2 and ROCK2 at greater than 70%. While HSD1400 also inhibited CLK4, the C1 methylated analog HSD1791 did not inhibit CLK4. Antitumor effects of the top compounds were evaluated and dose–response analysis indicated potent inhibition of renal cancer and leukemia cell growth. Immunoblotting results indicated that the top compounds induce DNA damage via upregulation of p-H2AX. Moreover, flow cytometry results demonstrated that the top compounds promote cell cycle arrest in the renal cancer cell line, Caki-1. qPCR and immunoblotting analysis upon HSD1791 dosing indicated suppression of cyclin D/Rb oncogenic pathway upon compound treatment. Conclusions: Novel boronic acid-containing pyrazolo[4,3-f]quinoline-based dual CLK/ROCK inhibitors were identified. The so-called “magic methylation” design approach was used to tune CLK selectivity. Additionally, the findings demonstrate potent in vitro anticancer activity of the lead candidates against renal cancer and leukemia. This adds to the growing list of boron-containing compounds that display biological activities.

Pharmaceuticals doi: 10.3390/ph17121659

Authors: Peter J. G. Remoto Keith C. Gordon Sara J. Fraser-Miller

Background/Objectives: Two fibre optic probes were custom designed to perform Raman and near-infrared spectroscopic measurements. Our long-term objective is to develop a non-destructive tool able to collect data in hard-to-access locations for real-time analysis or diagnostic purposes. This study evaluated the quantitative performances of Probe A and Probe B using model pharmaceutical tablets. Methods: Measurements were performed using pharmaceutical tablets containing hydroxyl propylcellulose, titanium dioxide (anatase), lactose monohydrate, and indomethacin (γ form). Material content and thickness of bilayer samples (samples consisting of a top layer and a bottom layer of differing materials) were also assessed using Probe A to evaluate its capabilities to collect sub-surface information. Principal component analysis and partial least squares regression models were using individual and fused data to evaluate the performances of the different probe configurations. Results: Hydroxymethyl cellulose (RP2=0.98, RMSEP = 2.27% w/w) and lactose monohydrate (RP2=0.97, RMSEP = 2.96% w/w) content were most effectively estimated by near-infrared spectroscopy data collected using Probe A. Titanium dioxide (RP2=0.99, RMSEP = 0.21% w/w) content was most effectively estimated using a combination of 785 nm Raman spectroscopy and near-infrared spectroscopy using Probe B. Indomethacin (RP2=0.97, RMSEP = 1.01% w/w) was best estimated using a low-level fused dataset collected using 0 mm, 2.5 mm, and 5.0 mm lateral offsets of 785 nm spatially offset Raman spectroscopy using Probe A. Conclusions: The different probe configurations were able to reliably collect data and demonstrated robust quantitative performances. These results highlight the advantage of using multiple techniques for analysing different structures.

Pharmaceuticals doi: 10.3390/ph17121658

Authors: Francisco Rivas García José Antonio García Sierra Maria-Isabel Valverde-Merino Maria Jose Zarzuelo Romero

Food supplements are used for a variety of purposes, one of which is weight reduction. As excess weight is a long-term condition, some supplements are expected to be used for long periods of time. The long-term use of these dietary supplements makes it highly likely that they will be combined with medications, increasing the risk of food supplement–drug interactions, which are not always known or disclosed, and can lead to serious health problems, as has been observed. This article discusses some of the compounds used as food supplements for weight reduction (green tea extract, Garcinia cambogia, chitosan, quercetin and resveratrol) and the interactions they may cause with some drugs such as: dextromethorphan, buspirone, diclofenac, irinotecan, 5-fluorouracil, cytochrome P450 inducers and inhibitors, statins, orlistat, warfarina, acenocoumarol, fluoxetine, valproate, quetiapine, carbamazepine. This information is expected to be useful for healthcare professionals to detect and intervene on food supplement–drug interactions to ensure the optimization of therapy and patient safety.

Pharmaceuticals doi: 10.3390/ph17121657

Authors: Dana Cucu

Background: The expression of the transient receptor potential 1 (TRPA1) gene is increased in many solid tumours, and its function relates to inflammation, oxidative stress or the presence of toxic substances. However, little is known about the correlation of clinical parameters with patients’ cancer stages, metastases and the degree of tumour infiltration by immune cells. Methods: We performed a bioinformatic analysis, using databases and public resources to investigate TRPA1 for many available samples. We used samples from the TCGA project and quantified the mRNA expression and survival analysis using TIMER, TIMER.2 and GEPIA. To analyse hypermethylation, a more extensive database was available from the UALCAN website. Results: We show that the TRPA1 gene is hypermethylated in many cancers. The high expression of TRPA1 is correlated with a better prognosis for several cancer types and correlates with cancer stage and metastasis, while in others the TRPA1 is pro-oncogenic. We also report the effect of TRPA1 expression in immune infiltrating cells. Moreover, the expression is linked to genes essential for inflammation, oxidative stress and cellular motility processes. Conclusions: Our study brings new insights into the regulation of TRPA1 expression in different tumours based on analysis provided by public databases, opening the possibility to further investigate the protein as a putative _target for cancer.

Pharmaceuticals doi: 10.3390/ph17121655

Authors: Fang Lv Xiaoling Cai Chu Lin Wenjia Yang Linong Ji

Background/Objectives: Type 2 diabetes and weight loss are associated with detrimental skeletal health. Incretin-based therapies (GLP-1 receptor agonists, and dual GIP/GLP-1 receptor agonists) are used clinically to treat diabetes and obesity. The potential effects of semaglutide and tirzepatide on bone metabolism in type 2 diabetic mice remain uncertain. Methods: Combined streptozotocin and high fat feeding were employed in female C57BL/6J mice to promote hyperglycemia. Mice were administered for 4 weeks with a saline vehicle (sc., once-daily), semaglutide (40 μg/kg/d, sc., every three days), or tirzepatide (10 nmol/kg, sc., once-daily). Bone strength was assessed by three-point bending. Femur microarchitecture was determined by micro-CT, and bone formation and resorption parameters were measured by histomorphometric analysis. Serum was collected to measure bone resorption (C-telopeptide fragments of type I collagen, CTX) and formation (procollagen type 1 N-terminal propeptide, P1NP) biomarkers, respectively. The expression of bone metabolism-related genes was evaluated in the bone using RT-PCR. Results: Glucose levels significantly reduced after 4 weeks of semaglutide and tirzepatide treatment (both p < 0.05) compared with vehicle treatment. Tirzepatide led to more weight loss than semaglutide. Compared to saline-treated diabetic mice, the mean femur length was shorter in the tirzepatide group. After treatment with tirzepatide or semaglutide, cortical bone and trabecular bone parameters did not change significantly compared to saline-treated diabetic mice, except that cortical thickness was lower in the semaglutide group compared to the saline group (p = 0.032). Though CTX and P1NP levels decreased, however, the change in CTX and P1NP levels did not differ among the four groups during the 4 weeks of treatment (all p > 0.05). Semaglutide affected RANKL and OPG mRNA expression and increased the ratio of OPG/RANKL. No significant difference was found in the quantity of Col1a1, RANKL, OPG, and RUNX2 between tirzepatide- and saline-treated diabetic mice. Conclusions: The 4-week treatment with semaglutide and tirzepatide had a neutral effect on bone mass compared with the controls, and most of the bone microarchitecture parameters were also comparable between groups in diabetic mice. A better understanding of incretin-based therapies on bone metabolism in patients with diabetes requires further evaluation in large clinical trials.

Pharmaceuticals doi: 10.3390/ph17121656

Authors: Lydia Whitham Mahdi Sheikh Markus W. Hollmann Marie-Odile Parat

Background/Objectives: Opium consumption was recently classified by the International Agency for Research on Cancer (IARC) monograph as carcinogenic to humans based on strong evidence for cancers of the larynx, lung, and urinary bladder, and limited evidence for cancers of the oesophagus, stomach, pancreas, and pharynx. This poses the question of a potential pro-cancer effect of pharmaceutical opioid analgesics. In vitro studies employing a variety of experimental conditions suggest that opioid alkaloids have proliferative or antiproliferative effects. We set out to reconcile this discrepancy and explore the hypothesis that opioids promote cancer cell proliferation in an organ-dependent fashion. Methods: Using strictly controlled conditions, we tested the effect of morphine on the proliferation of a series of human cancer cell lines isolated from organs where cancer risk was linked causally to opium consumption in human studies (i.e., lung, bladder, and larynx), or control organs where no link between cancer risk and opium consumption has been reported in human studies (i.e., breast, colon, prostate). Results: Our results showed a minimal effect on proliferation on any cell line and no trend supporting an organ-specific effect of morphine. Conclusions: This argues against a direct effect of opioids on tumour cell proliferation to support their organ-specific effect.

Pharmaceuticals doi: 10.3390/ph17121654

Authors: Hülya Tezel Yalçın Nadir Yalçın Michael Ceulemans Karel Allegaert

Background/Objectives: While breastfeeding is highly recommended, breastfed infants may be exposed to drugs by milk due to maternal pharmacotherapy, resulting in a risk of adverse drug events (ADE) or reactions (ADRs). The U.S. Food and Drug Administration (FDA) Adverse Event Reporting System (FAERS) is an online pharmacovigilance database, while the Drugs and Lactation Database (LactMed®) includes accurate and evidence-based information on levels of substances in breast milk and infant blood, and possible ADRs in nursing infants. We aimed to explore the FAERS database and compare ADE/ADR information patterns between both databases. Methods: The FAERS database was explored (29 July 2024) for ADEs related to drug exposure during lactation to determine annual trends, infant outcomes, and regions of reporting. The active pharmaceutical ingredients (APIs) associated with these ADEs were categorized based on the Anatomical Therapeutic Chemical (ATC, first level) classification. The top five APIs in each ATC group were explored in terms of the type of ADEs reported and compared to information in LactMed®. Results: In total, 2628 ADEs were obtained from the FAERS database, with increased reporting over time. In the FAERS database, 68.4% of the patients were under 2 months old, 5.5% had life threatening ADEs, and 3.6% died, while 84.70% of the cases were categorized as serious. Most ADEs were from North America (44.9%). Most drugs (50.9%) were nervous system drugs. The most frequent reported outcome was “other outcomes (without additional subdivision or information)” (58.2%), reflecting the diversity in outcomes reported. When related to the same drug, the FAERS database and LactMed® resource exhibited both similarities and differences in the types of reported ADE/ADR. Conclusions: The FAERS database is a useful tool to detect potential ADEs (rather sensitive), without ADR assessment, while LactMed® provides guidance driven by relevant ADRs (rather specific). The FAERS database is useful to obtain exploratory information about ADEs during lactation to increase the knowledge about drug safety during breastfeeding and the awareness of the possible risks in nursing infants, while LactMed® translates all available information into guidance.

Pharmaceuticals doi: 10.3390/ph17121653

Authors: Daniel E. K. Kabotso David Neglo Sarah E. Gaba Emmanuel K. Danyo Alberta D. Dayie Anastasia A. Asantewaa Fleischer C. N. Kotey Nicholas T. K. D. Dayie

Background: Antimicrobial resistance (AMR) has become precarious, warranting investments in antimicrobial discovery. Aim: To investigate the antibacterial activity of rosemary essential oil (REO), alone and in combination with selected conventional antibiotics. Methods: REO was subjected to antimicrobial susceptibility testing (including minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) determination) and investigation of anti-pre-biofilm and antibiofilm activities. Results: The phytochemical composition of the REO was eucalyptol (42.68%), bornanone (33.20%), endo-borneol (9.37%), α-terpeneol (7.95%), linalool (2.10%), bornyl acetate (1.81%), caryophyllene (1.09%), 4-terpeneol (0.94%), and anethole (0.87%). The antibacterial inhibition zones generally increased with increasing REO concentration (i.e., 10, 20, 50, 100, and 200 mg/mL). The MIC and MBC ranges of REO for all bacteria were 3.13–6.25 mg/mL and 3.12–12.5 mg/mL, respectively. The MICs (in µg/mL) of ciprofloxacin, chloramphenicol, streptomycin, tetracycline, and ampicillin, respectively, were Escherichia coli (0.98, 3.92, 1.96, 7.81, and 250), Klebsiella pneumoniae (1.25, 7.81, 125, 7.81, and 1000), MRSA (62.5, 7.81, 3.91, 7.81, and 250), Streptococcus mutans and Bacillus subtilis (125, 15.68, 250, 31.25, and 1000), Pseudomonas aeruginosa (125, 31.25, 500, 31.25, and 1000), and Salmonella Typhi (0.98, 15.68, 125, 1.96, and 1000). The MBC-MIC ratios of REO against all bacteria were in the range 1–2, indicating bactericidal effects. Mainly synergy (FICI = 0.16–0.37) was observed between REO and the conventional antibiotics. The IC50 values (in µg/mL) of REO against the bacteria, pre-biofilm vs. biofilm formation, were E. coli (1342.00 vs. 4.00), K. pneumoniae (106.00 vs. 3.00), MRSA (134.00 vs. 6.00), S. mutans (7259.00 vs. 7.00), B. subtilis (120.00 vs. 7.00), P. aeruginosa (4989.00 vs. 7.00), and S. Typhi (10.00 vs. 2.00). Conclusions: Rosemary essential oil had significant bactericidal effects on the bacteria tested, and its MIC and MBC values were low. Overall, it was synergistic with known conventional antibiotics and, thus, has encouraging prospects in combination therapy involving conventional antibiotics, even in the treatment of infections with multidrug-resistant bacteria, including biofilm-forming ones.

Pharmaceuticals doi: 10.3390/ph17121652

Authors: Naoufal El Hachlafi Amine Elbouzidi Amine Batbat Mohamed Taibi Mohamed Jeddi Mohamed Addi Hanae Naceiri Mrabti Kawtar Fikri-Benbrahim

Background/Objectives: Essential oils (EOs) from Citrus species have attracted attention for their diverse properties, including anti-inflammatory, antioxidant and cytotoxic effects, which address critical health challenges such as chronic diseases and skin disorders. Citrus sinensis (L.) Osbeck, which is a widely cultivated citrus fruit, is attracting increasing attention in the field of medicinal research due to its richness of limonene (comprising approximately 85–90% of the oil). This study investigates the chemical profile of CS-EO and biological activities of CS-EO and limonene. Methods and Results: This study used gas chromatography–mass spectrometry (GC-MS), confirming limonene as the predominant compound (70.15%) along with other minor constituents, including thujene (10.52%), myrcene (5.54%) and α-pinene (2.81%). The biological activities of CS-EO and limonene were examined, specifically focusing on their antioxidant, anti-inflammatory, cytotoxicity and dermatoprotective effects. Antioxidant potential was evaluated using DPPH, FRAP and beta-carotene assays, with CS-EO and limonene exhibiting comparable efficacy. Anti-inflammatory properties were assessed via inhibition assays of prostaglandin E2 (PGE2) and nitric oxide (NO) production, showing significant reductions in LPS-stimulated macrophages treated by CS-EO or limonene. Cytotoxicity testing on various cell lines indicated selective activity of the tested compounds, with low toxicity observed on human skin fibroblasts. Limonene and CS-EO were highly effective on HepG2 cellules, with IC50 values of 0.55 ± 0.01 µg/mL and 15.97 ± 1.20 µg/mL, respectively. Dermatoprotective effects were further confirmed using enzymes, where CS-EO and limonene showed remarkable inhibitory potential against elastase (IC50 of 65.72 ± 1.92 and 86.07 ± 1.53 µg/mL, respectively) and tyrosinase (IC50 of 102 ± 2.16 and 78.34 ± 1.15 µg/mL, respectively) enzymes compared to quercetin used as a standard (IC50 of 111.03 ± 0.1 and 124.22 ± 0.07 µg/mL, respectively). Conclusions: The findings of this study suggest the potential for the development of new therapeutic approaches based on CS-EO, which could be applicable in the pharmaceutical, cosmetic and nutraceutical fields and have protective benefits for skin health.

Pharmaceuticals doi: 10.3390/ph17121650

Authors: Abdo Meyiah Satya Kumar Avula Ahmed Al-Harrasi Eyad Elkord

Background: 3-O-acetyl-11-keto-β-boswellic acid (β-AKBA), a triterpene natural product, is one of the main natural products of Boswellia sacra resin (BSR) and has reported biological and immunomodulatory effects. 1H-1,2,3-triazole derivatives of β-AKBA (named 6a–6d) were synthesized from β-AKBA. The 1H-1,2,3-triazole compounds are also known to have a wide range of biological and pharmacological properties as demonstrated by in vitro and in vivo studies. This study aimed to investigate the effects of these 1H-1,2,3-triazole derivatives of β-AKBA on human T-cell proliferation and activation. Methods: PBMCs isolated from healthy donors were activated by anti-CD3/CD28 monoclonal antibodies in the presence of β-AKBA (1) or 1H-1,2,3-triazole derivatives of β-AKBA or DMSO controls. Results: We found that similar to the parent compound β-AKBA (1), derivatives 6a, 6b, and 6d significantly inhibited T-cell expansion/proliferation and reduced the levels of CD25 activation marker on CD4+ and CD8+ T cells without exerting significant cytotoxic effects on T-cell viability at a concentration of 25 µM. However, compound 6c further inhibited T-cell expansion/proliferation and CD25 expression, but had a significant cytotoxic effect on cell viability at similar concentrations of 25 µM. Conclusions: These findings demonstrate the immunoinhibitory effects of β-AKBA (1) and its corresponding triazole derivatives on T-cell proliferation and activation, highlighting the promising therapeutic potential of these compounds in T-cell-mediated diseases.

Pharmaceuticals doi: 10.3390/ph17121651

Authors: Charles W. Emala Tarnjot K. Saroya Yuqi Miao Shuang Wang Shengmin Sang Emily A. DiMango

Background/Objective: A significant number of individuals with asthma have poorly controlled daily symptoms and utilize dietary supplements such as ginger in a quest for improved symptom control; however, its effectiveness at improving the control of symptoms is unproven. We questioned whether low-dose oral ginger would improve subjective and objective measurements of asthma control in mild-to-moderate asthmatics. Methods: We performed a randomized, placebo-controlled, double-blinded study of a low dose (1 g twice daily) of a dietary supplement of ginger in 32 mild-to-moderate uncontrolled asthmatics over a 2-month trial period while maintaining daily conventional asthma therapies. The planned primary outcomes included an increased tolerance to inhaled methacholine and decreased concentrations of fractional excretion of exhaled nitric oxide (FeNO). Secondary planned outcomes included measurements of asthma control by the Asthma Control Test (ACT), a 2-week symptom recall test, and the Juniper mini Asthma Quality of Life Questionnaire (AQLQ), and blood eosinophils and asthma-associated cytokines. Results: Exhaled nitric oxide or blood eosinophils were not changed by oral ginger. However, three different measures of asthma symptom control were improved by the 28-day time point of oral ginger. Asthma-associated serum cytokines (IL-13 and IL-17A) were modulated by oral ginger. Conclusions: This is the first demonstration that a small daily dose of a dietary supplement of ginger may improve asthma symptoms and reduce inflammation in human asthmatics. These findings support the need for additional studies using larger doses of ginger in specific endotypes of asthmatics that may identify a novel therapeutic for asthma.

Pharmaceuticals doi: 10.3390/ph17121649

Authors: Kazim Sahin Ahmet Kayhan Korkusuz Emre Sahin Cemal Orhan Besir Er Abhijeet Morde Muralidhara Padigaru Ertugrul Kilic

Background/Objectives: With increasing interest in plant-based compounds that can enhance sleep quality without the side effects of caffeine, Alpinia galanga (AG) has emerged as a promising herbal supplement for improving mental alertness. This study assessed the impact of water-soluble AG extract on sleep quality; the activity of GABAergic, glutamatergic, and serotonergic receptors; and concentrations of dopamine and serotonin in the brains of mice. Methods: The study employed two experimental models using BALB/c mice to examine the impact of pentobarbital-induced sleep and caffeine-induced insomnia. In the first model, a set of 20 mice was assigned to four groups to assess the effects of pentobarbital (42 mg/kg) or pentobarbital with AG extract on sleep induction, with observations made 45 min post-administration. In the second model, 20 mice were divided into four groups to evaluate the impact of caffeine (25 mg/kg) alone or caffeine with varying doses of AG extract (61.25 or 205.50 mg/kg administered orally) on brain activity along with additional analyses on receptor proteins and neurotransmitters. Results: A higher dose of AG extract (205.50 mg/kg) significantly increased total deep sleep duration compared to the caffeine group (p < 0.0001). Furthermore, this dose extended sleep latency and suppressed GABAergic and glutamatergic receptor activity compared to the lower AG dose (p < 0.05). Additionally, the 205.50 mg/kg dose elevated serotonin and dopamine levels compared to caffeine (p < 0.0001), suggesting improved sleep quality alongside enhanced wakefulness. Conclusions: Our data indicate that a higher dose of AG extract improved sleep latency and duration by regulating GABAergic and glutamatergic receptors through the GABAergic/serotonergic pathway in mice.

Pharmaceuticals doi: 10.3390/ph17121648

Authors: Aleksandar Jovalekic Santiago Bullich Núria Roé-Vellvé Guilherme Domingues Kolinger Lorelei R. Howard Floriana Elsholz Mariana Lagos-Quintana Beatriz Blanco-Rodriguez Esther Pérez-Martínez Rossella Gismondi Audrey Perrotin Marianne Chapleau Richard Keegan Andre Mueller Andrew W. Stephens Norman Koglin

Florbetaben (FBB) is a radiopharmaceutical approved by the FDA and EMA in 2014 for the positron emission tomography (PET) imaging of brain amyloid deposition in patients with cognitive impairment who are being evaluated for Alzheimer’s disease (AD) or other causes of cognitive decline. Initially, the clinical adoption of FBB PET faced significant barriers, including reimbursement challenges and uncertainties regarding its integration into diagnostic clinical practice. This review examines the progress made in overcoming these obstacles and describes the concurrent evolution of the diagnostic landscape. Advances in quantification methods have further strengthened the traditional visual assessment approach. Over the past decade, compelling evidence has emerged, demonstrating that amyloid PET has a strong impact on AD diagnosis, management, and outcomes across diverse clinical scenarios, even in the absence of amyloid-_targeted therapies. Amyloid PET imaging has become essential in clinical trials and the application of new AD therapeutics, particularly for confirming eligibility criteria (i.e., the presence of amyloid plaques) and monitoring biological responses to amyloid-lowering therapies. Since its approval, FBB PET has transitioned from a purely diagnostic tool aimed primarily at excluding amyloid pathology to a critical component in AD drug development, and today, it is essential in the diagnostic workup and therapy management of approved AD treatments.

Pharmaceuticals doi: 10.3390/ph17121646

Authors: Asma N. Alsaleh Ibrahim M. Aziz Reem M. Aljowaie Rawan M. Alshalan Noorah A. Alkubaisi Mourad A. M. Aboul-Soud

Artemisia absinthium L., is a plant with established pharmacological properties, but the A. absinthium root extract (AARE) remains unexplored. The aim of this study was to examine the chemical composition of AARE and assess its biological activity, which included antidiabetic, antibacterial, anticancer, and antioxidant properties. GC-MS was used to analyze the chemical components. The antioxidant activity of the total phenolic and flavonoid content was evaluated. Antibacterial activity and cytotoxic effects were identified. Enzyme inhibition experiments were performed to determine its antidiabetic potential. Molecular docking was utilized to evaluate the potential antioxidant, antibacterial, and anticancer activities of the compounds from AARE using Maestro 11.5 from the Schrödinger suite. AARE exhibited moderate antioxidant activity in DPPH (IC50: 172.41 ± 3.15 μg/mL) and ABTS (IC50: 378.94 ± 2.18 μg/mL) assays. Cytotoxicity tests on MCF-7 and HepG2 cancer cells demonstrated significant anticancer effects, with IC50 values of 150.12 ± 0.74 μg/mL and 137.11 ± 1.33 μg/mL, respectively. Apoptotic studies indicated an upregulation of pro-apoptotic genes (caspase-3, 8, 9, Bax) and a downregulation of anti-apoptotic markers (Bcl-2 and Bcl-Xl). AARE also inhibited α-amylase and α-glucosidase, suggesting potential antidiabetic effects, with IC50 values of 224.12 ± 1.17 μg/mL and 243.35 ± 1.51 μg/mL. Antibacterial assays revealed strong activity against Gram-positive bacteria. Molecular docking and pharmacokinetic analysis identified promising inhibitory effects of key AARE compounds on NADPH oxidase, E. coli Gyrase B, and Topoisomerase IIα, with favorable drug-like properties. These findings suggest AARE’s potential in treating cancer, diabetes, and bacterial infections, warranting further in vivo and clinical studies.

Pharmaceuticals doi: 10.3390/ph17121647

Authors: Huanhuan Liu Xiaojun Zhang Yue Pan Jingfeng Zhang Hui Wen Cong Zhang Xiaodan Xu Guangyu Ma Ruimin Wang Jinming Zhang

Objective: Currently, 68Ga-labeled somatostatin analogs (SSAs) are the most commonly used imaging agents for patients with neuroendocrine tumors (NETs) in clinical practice, demonstrating good results in tumor diagnosis. For applications in peptide receptor radionuclide therapy (PRRT), _targeted drugs should have high tumor uptake and prolonged tumor retention time. To enhance the uptake and retention of tracers in NETs, our goal is to design a 68Ga-labeled heterodimer for optimizing pharmacokinetics and assess whether this form is more efficacious than its monomeric equivalents. Methods: Using the somatostatin analog TATE and quinoline-based compound FAPI-46 as raw materials, we designed and synthesized 68Ga-labeled TATE-46. The labeling efficiency and stability were verified by Radio-HPLC. The receptor binding properties and tumor _targeting were examined both in vitro and in vivo by using NCI-H727 (SSTR2/FAP, positive) and Mc38 (SSTR2/FAP, negative) cell lines and tumor-bearing mouse models. Preclinical evaluation was performed through cell uptake, pharmacokinetics, Micro PET, and biodistribution studies, and the results were compared with [68Ga]Ga-DOTA-TATE and [68Ga]Ga -FAPI-46. Immunohistochemistry and HE staining were performed on tumor tissues from tumor-bearing mice for further validation. Results: [68Ga]Ga-TATE-46 showed comparable SSTR2 and FAP _targeting ability to monomeric TATE and FAPI-46 in cell uptake and PET imaging studies. [68Ga]Ga-TATE-46 exhibited significantly higher uptake in NCI-H727 (SSTR2/FAP, positive) tumors compared to [68Ga]Ga-DOTA-TATE (p < 0.001) and [68Ga]Ga-FAPI-46 (p < 0.001). No increased uptake of [68Ga]Ga-TATE-46 was observed in MC38 tumors (SSTR2/FAP, negative). Additionally, excess DOTA-TATE and/or unlabeled FAPI-46 significantly blocked the uptake of [68Ga]Ga-TATE-46 in NCI-H727 tumors (p < 0.001), confirming its dual-receptor _targeting characteristics. The ex vivo biodistribution, immunofluorescence and immunohistochemistry results were in line with the in vivo imaging findings. Conclusion: Compared with 68Ga-labeled FAPI-46 and DOTA-TATE mono-specific tracers, the dual-_target tracer [68Ga]Ga-TATE-46 improves tumor uptake, extends tumor retention, and enhances pharmacokinetics. It is an effective probe for non-invasive detection of tumors expressing FAP and SSTR2, and it is worth further studying its application in the expression of sstr2 and FAP-related tumors.

Pharmaceuticals doi: 10.3390/ph17121645

Authors: Reetuparna Acharya Pran Kishore Deb Katharigatta N. Venugopala Shakti Prasad Pattanayak

Background: Breast cancer influences more than 2 million women worldwide annually. Since apoptotic dysregulation is a cancer hallmark, _targeting apoptotic regulators encompasses strategic drug development for cancer therapy. One such class of apoptotic regulators is inhibitors of apoptosis proteins (IAP) which are a class of E3 ubiquitin ligases that actively function to support cancer growth and survival. Methods: The current study reports design, synthesis, docking analysis (based on binding to IAP-BIR3 domains), anti-proliferative and anti-tumor potential of the azomethine derivative, 1-(4-chlorophenyl)-N-(4-ethoxyphenyl)methanimine (BCS3) on breast cancer (in vitro and in vivo) and its possible mechanisms of action. Results: Strong selective cytotoxic activity was observed in MDA-MB-231, MCF-7, and MDA-MB-468 breast cancer cell lines that exhibited IC50 values, 1.554 µM, 5.979 µM, and 6.462 µM, respectively, without affecting normal breast cells, MCF-10A. For the evaluation of the cytotoxic potential of BCS3, immunofluorescence, immunoblotting, and FACS (apoptosis and cell cycle) analyses were conducted. BCS3 antagonized IAPs, thereby causing MDM2-p53 and Bcl-2-Caspase-mediated intrinsic and extrinsic apoptosis. It also modulated p53 expression causing p21-CDK1/cyclin B1-mediated cell cycle arrest at S and G2/M phases. The in vitro findings were consistent with in vivo findings as observed by reduced tumor volume and apoptosis initiation (TUNEL assay) by IAP downregulation. BCS3 also produced potent synergistic effects with doxorubicin on tumor inhibition. Conclusions: Having witnessed the profound anti-proliferative potential of BCS3, the possible adverse effects related to anti-cancer therapy were examined following OECD 407 guidelines which confirmed its systemic safety profile and well tolerability. The results indicate the promising effect of BCS3 as an IAP antagonist for breast cancer therapy with fewer adverse effects.

Pharmaceuticals doi: 10.3390/ph17121644

Authors: João Luís Q. Simei José Diogo R. Souza João Francisco Pedrazzi Francisco S. Guimarães Alline Cristina Campos Antônio Zuardi Jaime Eduardo C. Hallak José Alexandre S. Crippa

Background: Emerging evidence supports cannabidiol (CBD) as a promising therapeutic compound for various health conditions, despite its approval as a medication (product for medical purposes) remaining restricted to a limited range of clinical indications. Simultaneously, the regulation of cannabis-derived products for medicinal and recreational use has expanded their global market availability to meet local community demands. This scenario presents a complex challenge for clinicians, researchers, and industry, as the global appeal of therapeutic uses of CBD is growing more rapidly than the scientific evidence supporting its safety and effectiveness. Outcomes: A narrative review was conducted to discuss the best evidence regarding the pharmacological profile of CBD, its efficacy, and safety within the context of regulation and perspectives on the development of new cannabinoid-based drugs. Key articles addressing the various facets of this issue were selected for comprehensive analysis. Conclusions: Clinicians and researchers may face unique challenges in understanding the pharmacological profile of CBD and the prospects for developing its clinical indications, given the heterogeneity of clinical terminologies and the quality and composition of cannabis-based medical products available on the market. More basic and clinical research that complies with regulatory agencies’ testing guidelines, such as good manufacturing practices (GMPs), good laboratory practices (GLPs), and good clinical practices (GCPs), is needed to obtain approval for CBD or any other cannabinoid as a therapeutic for broader clinical indications.

Pharmaceuticals doi: 10.3390/ph17121643

Authors: Charlotte Day John-Paul Silva Rebecca Munro Brice Mullier Véronique Marie André Christian Wolff Gary J. Stephens Angela Bithell

Background/Objectives: Recent studies, typically using patient cerebrospinal fluid (CSF), have suggested that different autoantibodies (Aabs) acting on their respective receptors, may underlie neuropsychiatric disorders. The GluN1 (NR1) subunit of the N-methyl-D-aspartate receptor (NMDAR) has been identified as a _target of anti-NMDAR Aabs in a number of central nervous system (CNS) diseases, including encephalitis and autoimmune epilepsy. However, the role or the nature of Aabs responsible for effects on neuronal excitability and synaptic plasticity is yet to be established fully. Methods: Peptide immunisation was used to generate Aabs against selected specific GluN1 extracellular sequences based on patient-derived anti-NMDAR Aabs that have been shown to bind to specific regions within the GluN1 subunit. ‘Protein A’ purification was used to obtain the total IgG, and further peptide purification was used to obtain a greater percentage of NMDAR-_target specific IgG Aabs. The binding and specificity of these anti-NMDAR Aabs were determined using a range of methodologies including enzyme-linked immunosorbent assays, immunocytochemistry and immunoblotting. Functional effects were determined using different in vitro electrophysiology techniques: two-electrode voltage-clamps in Xenopus oocytes and measures of long-term potentiation (LTP) in ex vivo hippocampal brain slices using multi-electrode arrays (MEAs). Results: We show that anti-NMDAR Aabs generated from peptide immunisation had specificity for GluN1 immunisation peptides as well as _target-specific binding to the native protein. Anti-NMDAR Aabs had no clear effect on isolated NMDARs in an oocyte expression system. However, peptide-purified anti-NMDAR Aabs prevented the induction of LTP at Schaffer collateral-CA1 synapses in ex vivo brain slices, consistent with causing synaptic NMDAR hypofunction at a network level. Conclusions: This work provides a solid basis to address outstanding questions regarding anti-NMDAR Aab mechanisms of action and, potentially, the development of therapies against CNS diseases.

Pharmaceuticals doi: 10.3390/ph17121642

Authors: Nikolay Kalitin Natalia Koroleva Anna Lushnikova Maria Babaeva Nadezhda Samoylenkova Ekaterina Savchenko Galina Smirnova Yulia Borisova Alexander Kostarev Aida Karamysheva Galina Pavlova

Background/Objectives: Indolo[2,3-a]pyrrolo[3,4-c]carbazole scaffold is successfully used as an efficient structural motif for the design and development of different antitumor agents. In this study, we investigated the anti-glioblastoma therapeutic potential of glycosylated indolocarbazole analog LCS1269 utilizing in vitro, in vivo, and in silico approaches. Methods: Cell viability was estimated by an MTT assay. The distribution of cell cycle phases was monitored using flow cytometry. Mitotic figures were visualized by fluorescence microscopy. Quantitative RT-PCR was used to evaluate the gene expression. The protein expression was assessed by Western blotting. Molecular docking and computational ADMET were approved for the probable protein _target simulations and predicted pharmacological assessments, respectively. Results: Our findings clearly suggest that LCS1269 displayed a significant cytotoxic effect against diverse glioblastoma cell lines and patient-derived glioblastoma cultures as well as strongly suppressed xenograft growth in nude mice. LCS1269 exhibited more potent anti-proliferative activity toward glioblastoma cell lines and patient-derived glioblastoma cultures compared to conventional drug temozolomide. We further demonstrated that LCS1269 treatment caused the severe G2 phase arrest of cell cycle in a dose-dependent manner. Mechanistically, we proposed that LCS1269 could affect the CDK1 activity both by _targeting active site of this enzyme and indirectly, in particular through the modulation of the Wee1/Myt1 and FOXM1/Plk1 signaling pathways, and via p21 up-regulation. LCS1269 also showed favorable pharmacological characteristics in in silico ADME prediction in comparison with staurosporine, rebeccamycin, and becatecarin as reference drugs. Conclusions: Further investigations of LCS1269 as an anti-glioblastoma medicinal agent could be very promising.

Pharmaceuticals doi: 10.3390/ph17121641

Authors: Antonia Dimitrakopoulou-Strauss Leyun Pan Christos Sachpekidis

Molecular imaging is a growing field, driven by technological advances, such as the improvement of PET-CT scanners through the introduction of digital detectors and scanners with an extended field of view, resulting in much higher sensitivity and a variety of new specific radiopharmaceuticals that allow the visualization of specific molecular pathways and even theragnostic approaches. In oncology, the development of dedicated tracers is crucial for personalized therapeutic approaches. Novel peptides allow the visualization of many different _targets, such as PD-1 and PD-L1 expression, chemokine expression, HER expression, T-cell imaging, microenvironmental imaging, such as FAP imaging, and many more. In this article, we review recent advances in the development of non-[18F]FDG PET radiopharmaceuticals and their current clinical applications in oncology, as well as some future aspects.

Pharmaceuticals doi: 10.3390/ph17121640

Authors: Susan Cole Henry Pertinez Andrew S. Butler Essam Kerwash Swati Bhat Eman El-Khateeb Andrew Owen

Background: The development of long-acting products of a characterized drug substance is of great interest. It is possible to support the development of these products with available clinical data by matching the exposure to a predefined bracket of a minimal concentration for efficacy and a maximal concentration for safety. This bracketing approach would cut down on the time and cost of new long-acting contraceptive products progressing to market. The current study describes the assessment of the data available to support a bracketing approach to conclude comparable levels of efficacy and safety for a postulated novel long-acting reversible contraceptive (LARC) product of levonorgestrel. Methods: Literature evidence of levonorgestrel efficacy, as quantified by the Pearl Index, was utilized and modeled by incorporating three LARC products for the estimation of a minimal concentration required for efficacy. Further literature was reviewed to quantify the maximal concentration required to ensure product safety. Additionally, a review of the regulatory precedence for the approach was conducted using European and UK databases. Results: There was a reasonable definition of the minimal concentrations for efficacy where the _target concentrations of levonorgestrel were in the range of 200–400 pg/mL. Maximum concentrations for safety were less well defined. Although regulatory guidance supports the bracketing approach, there is little precedence for licensing new products based on pharmacokinetic data only, despite much reduced clinical and non-clinical packages being evidenced. Conclusions: Understanding of the exposure response is not currently considered sufficient to support a bracketing approach for a new levonorgestrel product. If additional safety data are established, current regulations may allow for a reduced application package. Additional work is needed to support the approach, and this could utilize the wealth of information in real-world datasets combined with systems models.

Pharmaceuticals doi: 10.3390/ph17121639

Authors: Wei Zhu Xiaoyong Zhang Dong Wang Qinghua Yao Guang-Lei Ma Xiaohui Fan

The human gastrointestinal microbiota plays a vital role in maintaining host health and preventing diseases, prompting the creation of simulators to replicate this intricate system. The Simulator of the Human Intestinal Microbial Ecosystem (SHIME®), a multicompartment dynamic simulator, has emerged as a pivotal in vitro model for studying the interactions and interferences within the human gut microbiota. The continuous and real-time monitoring hallmarks, along with the programmatically flexible setup, bestow SHIME® with the ability to mimic the entire human intestinal ecosystem with high dynamics and stability, allowing the evaluation of various treatments on the bowel microbiota in a controlled environment. This review outlines recent developments in SHIME® systems, including the M-SHIME®, Twin-SHIME®, Triple-SHIME®, and Toddle SHIME® models, highlighting their applications in the fields of food and nutritional science, drug development, gut health research, and traditional Chinese medicine. Additionally, the prospect of SHIME® integrating with other advanced technologies is also discussed. The findings underscore the versatility of SHIME® technology, demonstrating its significant contributions to current gut ecosystem research and its potential for future innovation in microbiome-related fields.

Pharmaceuticals doi: 10.3390/ph17121638

Authors: Yaswanth Kuthati Chih-Shung Wong

Background: Morphine analgesic tolerance (MAT) limits the clinical application of morphine in the management of chronic pain. IIK7 is a melatonin type 2 (MT2) receptor agonist known to have antioxidant properties. Oxidative stress is recognized as a critical factor in MAT. This study sought to assess the impact of IIK7 on the progression of MAT and its potential to reverse pre-existing MAT. Methods: Wistar rats underwent partial sciatic nerve transection (PSNT) surgery to induce neuropathic pain (NP). Seven days post nerve transection, we implanted an intrathecal (i.t.) catheter and linked it to an osmotic pump. Rats were randomly divided into the following groups: sham-operated/vehicle, PSNT/vehicle, PSNT/IIK7 50 ng/h, PSNT/MOR 15 g/h, and PSNT/MOR 15 g + IIK7 50 ng/h. We implanted two i.t. catheters for drug administration and the evaluation of the efficacy of IIK7 in reversing pre-established MAT. We linked one to an osmotic pump for MOR or saline continuous i.t. infusion. On the 7th day, the osmotic pump was disconnected, and 50 μg of IIK7 or the vehicle was administered through the second catheter. After 3 h, 15 μg of MOR or saline was administered, and the animal behavior tests were performed. We measured the levels of mRNA for Nrf2 and HO-1, pro-inflammatory cytokines (PICs), and the microglial and astrocyte activation in the spinal cord. Results: The co-administration of IIK7 with MOR delayed MAT development in PSNT rats by restoring Nrf2 and HO-1 while also inhibiting the microglial-cell and astrocyte activation, alongside the suppression of PICs. Additionally, a single injection of high-dose 50 μg IIK7 was efficient in restoring MOR’s antinociception in MOR-tolerant rats. Conclusions: Our results indicate that the co-infusion of ultra-low-dose IIK7 can delay MAT development and a high dose can reverse pre-existing MAT.

Pharmaceuticals doi: 10.3390/ph17121637

Authors: Yingyao Wu Jianxin Cai Hanhe Liu Chan Li Qingfa Tang Yuan-Wei Zhang

Background: Albizia julibrissin Durazz. is one of the most popular herbs used for depression treatment, but the molecular basis for its mechanism of action has not been fully addressed. Previously, we isolated and identified two lignan glycoside derivatives that were shown to noncompetitively inhibit serotonin transporter (SERT) activity but with a relatively low inhibitory potency compared with those of conventional antidepressants. Methods: We characterized the pharmacological profile of the parental compound of these previously isolated lignan glycosides, (-)-syringaresinol (SYR), in inhibiting SERT by using biochemical, pharmacological, and behavioral approaches. Results: SYR, as a potent inhibitor, decreases SERT Vmax but with little change in Km for its fluorescent substrate. SYR was shown to block the conformational conversion essential for substrate transport by stabilizing SERT in an outward-open and inward-closed conformation. In addition, our molecular docking and biochemical validation demonstrated that SYR binds to an allosteric site in SERT and noncompetitively inhibits SERT transport and binding activity. Furthermore, administration of SYR was indicated to exert an antidepressant-like activity and to effectively attenuate chronic unpredictable mild stress (CUMS)-induced abnormalities in behaviors and synaptic protein expression in depressive animal models. Conclusions: This study not only provides molecular insights into the mechanism of action of A. julibrissin in the treatment of depression, but also opens up the possibility of development of a novel class of allosteric site-_targeted therapeutic agents with an underlying mechanism of action different from that of conventional antidepressants.

Pharmaceuticals doi: 10.3390/ph17121636

Authors: Xianghui Chen Zanwen Zuo Xianbin Li Qizhang Li Lei Zhang

Background/Objectives: Breast cancer is the second most common malignancy worldwide and poses a significant threat to women’s health. However, the prognostic biomarkers and therapeutic _targets of breast cancer are unclear. A prognostic model can help in identifying biomarkers and _targets for breast cancer. In this study, a novel prognostic model was developed to optimize treatment, improve clinical prognosis, and screen potential phosphoglycerate kinase 1 (PGK1) inhibitors for breast cancer treatment. Methods: Using data from the Gene Expression Omnibus (GEO) database, differentially expressed genes (DEGs) were identified in normal individuals and breast cancer patients. The biological functions of the DEGs were examined using bioinformatics analysis. A novel prognostic model was then constructed using the DEGs through LASSO and multivariate Cox regression analyses. The relationship between the prognostic model, survival, and immunity was also evaluated. In addition, virtual screening was conducted based on the risk genes to identify novel small molecule inhibitors of PGK1 from Chemdiv and _targetmol libraries. The effects of the potential inhibitors were confirmed through cell experiments. Results: A total of 230 up- and 325 down-regulated DEGs were identified in HER2, LumA, LumB, and TN breast cancer subtypes. A new prognostic model was constructed using ten risk genes. The analysis from The Cancer Genome Atlas (TCGA) indicated that the prognosis was poorer in the high-risk group compared to the low-risk group. The accuracy of the model was confirmed using the ROC curve. Furthermore, functional enrichment analyses indicated that the DEGs between low- and high-risk groups were linked to the immune response. The risk score was also correlated with tumor immune infiltrates. Moreover, four compounds with the highest score and the lowest affinity energy were identified. Notably, D231-0058 showed better inhibitory activity against breast cancer cells. Conclusions: Ten genes (ACSS2, C2CD2, CXCL9, KRT15, MRPL13, NR3C2, PGK1, PIGR, RBP4, and SORBS1) were identified as prognostic signatures for breast cancer. Additionally, results showed that D231-0058 (2-((((4-(2-methyl-1H-indol-3-yl)-1,3-thiazol-2-yl)carbamoyl)methyl)sulfanyl)acetic acid) may be a novel candidate for treating breast cancer.

Pharmaceuticals doi: 10.3390/ph17121635

Authors: Sodiq Kolawole Lawal Samuel Oluwaseun Olojede Babatunde Adebola Alabi Kafalotse Sylvia Dithole Samuel Thopho Matula Edwin Coleridge Naidu Carmen Olivia Rennie Onyemaechi Okpara Azu

Adverse complications like metabolic disorders, neurotoxicity, and low central nervous system (CNS) penetration are associated with the long-term use of tenofovir disoproxil fumarate (TDF). Therefore, some modifications are required to enhance neurological functions using silver nanoparticles (AgNPs). This study aimed to evaluate the neuroprotective impact of silver nanoparticles (AgNPs)-conjugated TDF as AgNPs-TDF on the hippocampal microanatomy and some neuro-biomarkers of diabetic rats. Forty-two male Sprague-Dawley rats, with an average weight of 250 ± 13 g, were divided into non-diabetic and diabetic groups. They were further divided into 3 groups each (n = 7): non-diabetic control (NC), non-diabetic + TDF (NTF), and non-diabetic + TDF + silver nanoparticles (NTS), as well as diabetic control (DC), diabetic + TDF (DTF), and diabetic + TDF + silver nanoparticles (DTS). The characterization of AgNPs-TDF was assessed, and the conjugates were administered to the diabetic rats, followed by behavioral testing and biochemical, immunohistochemical, and microanatomy analyses of the hippocampus. The results showed that the administration of AgNPs-TDF significantly reduced the blood glucose level, malondialdehyde (MDA), and inflammatory biomarker concentrations in DTS compared with the DTF and DC groups. Furthermore, AgNPs-TDF administration significantly increased the levels of tissue superoxide dismutase (SOD), reduced glutathione (GSH), and insulin-like growth factor-1 in DTS compared with the DTF and DC groups. In addition, the DTS group revealed a monomorphic pattern of dark-stained neuronal nuclei similar to the control group and showed neuroprotective effects on hippocampal microanatomy compared with the DTF group. This study shows that AgNPs-TDF restores various alterations in the hippocampus and improves cognitive functions in diabetic rats.

Pharmaceuticals doi: 10.3390/ph17121634

Authors: Qiang He Jie Wang Jingjing Li Wenchao Yang

Background/Objective: Bee pollen, a rich nutritional food, was employed to develop a raw material for skin whitening. Methods: The polyphenol profile and antioxidant, antityrosinase, and anti-melanogenesis activities of the ethanol extracts of five species of bee pollens (EEBPs) were determined. Results: The results showed that there were a total of 121 phenolic compounds in these EEBPs. Each type of bee pollen had unique substances. The best anti-melanogenesis activity was observed for sunflower EEBP, about 25% at a concentration of 25 μg/mL BEEP. The anti-melanogenesis activities of EEBPs from high to low were sunflower, apricot, camellia, rapeseed, and lotus EEBPs. The anti-melanogenesis activity in B16F10 cells was positively correlated with the antityrosinase activity and total phenol content, with coefficients of 0.987 and 0.940. The Kyoto Encyclopedia of Genes and Genomes enrichment analysis results of un_targeted proteomics revealed that sunflower EEBP inhibited melanogenesis in B16F10 cells by reducing the expression of the proteins MAP2K1, NFKB2, RELB, RPS6KA3, CASP3, TRAF6, MAP2K5, MAPKAPK3, STRADA, CCNA2, and FASN involved in the cAMP, MAPK, and TNF signaling pathways, even though these pathways were not significantly different from the control group. Conclusions: The sunflower EEBP has high inhibition effect on melanogenesis than other species EEBPs. The results provide a basis for the future industrial development of a raw material for skin whitening.

Pharmaceuticals doi: 10.3390/ph17121633

Authors: Chutintorn Somnin Joseph Chamieh Laurent Leclercq Christelle Medina Olivier Rousseaux Hervé Cottet

Background: Gadolinium-based contrast agents (GBCA) are widely used in magnetic resonance imaging (MRI) to enhance image contrast by interacting with water molecules, thus improving diagnostic capabilities. However, understanding the residual accumulation of GBCA in tissues after administration remains an area of active research. This highlights the need for advanced analytical techniques capable of investigating interactions between GBCAs and biopolymers, such as type I collagen, which are abundant in the body. Objective: This study explores the interactions of neutral and charged GBCAs with type I collagen under physiological pH conditions (pH 7.4) using Taylor dispersion analysis (TDA) and frontal analysis continuous capillary electrophoresis (FACCE). Methods: Collagen from bovine achilles tendon was ground using a vibratory ball mill to achieve a more uniform particle size and increased surface area. Laser granulometry was employed to characterize the size distributions of both raw and ground collagen suspensions in water. TDA was used to assess the hydrodynamic radius (Rh) of the soluble collagen fraction present in the supernatant. Results: From the TDA and FACCE results, it was shown that there were no significant interactions between the tested GBCAs and either the ground collagen or its soluble fraction at pH 7.4. Interestingly, we also observed that collagen interacts with filtration membranes, indicating that careful selection of membrane material, or the absence of filtration in the experimental protocol, is essential in interaction studies involving collagen. Conclusion: These findings bring valuable insights into the behavior of GBCAs in biological systems with potential implications for clinical applications.

Pharmaceuticals doi: 10.3390/ph17121632

Authors: Ameen Ali Abu-Hashem Sami A. Al-Hussain

Background: Recently, pyrido[2,3-d] pyrimidine, triazolopyrimidine, thiazolopyrimidine, quinoline, and pyrazole derivatives have gained attention due to their diverse biological activities, including antimicrobial, antioxidant, antitubercular, antitumor, anti-inflammatory, and antiviral effects. Objective: The synthesis of new heterocyclic compounds including 5-quinoline-pyrido[2,3-d] pyrimidinone (1–2, 4, 6–7), 6-quinoline-pyrido[2,3-d]thiazolo[3,2-a]pyrimidinone (3, 5, 8–10), 1,2,4-triazole-6-quinoline-pyrido[2,3-d]thiazolo[3,2-a]pyrimidinone (11–13), and pyrido[2,3-d]thiazolo[3,2-a]pyrimidine-ethyl-(pyridine)-9-thiaazabenzo[cd]azulenone (14) derivatives was performed with high yields while evaluating antimicrobial activities. Methods: A new series of quinoline-pyrido[2,3-d]thiazolo[3,2-a]pyrimidine derivatives were prepared using a modern style and advanced technology, resulting in high yields of these new compounds. Various reagents were utilized, specifically tailored to the production needs of each compound, through reactions that included alkylation, addition, condensation, acylation, the formation of Schiff bases, and intramolecular cyclization. Results: The chemical structures of the new compounds were determined using spectroscopy analyses, including IR, NMR, and MS, achieving good yields ranging from 68% to 90% under mild conditions in a regular system. All compounds were tested for in vitro antimicrobial activity and compared to standard drugs, specifically cefotaxime sodium and nystatin. The results showed that compounds 10 to 14 exhibited excellent antimicrobial activity, with a minimum inhibitory concentration (MIC) of 1 to 5 µmol/mL, compared to that of the standard drugs, which had MIC values of 1 to 3 µmol/mL. Furthermore, molecular docking studies were conducted to explore the interactions of specific compounds with antimicrobial _target proteins. The findings revealed that compounds 10 to 14 displayed significant binding energies, with ΔG values ranging from −7.20 to −11.70 kcal/mol, indicating effective binding to the active sites of antimicrobial protein receptors. Conclusions: The SAR study confirmed a relationship between antimicrobial activity and the tested compounds. Molecular docking demonstrated that compounds 10, 11, 12, 13, and 14 exhibited significant binding energy, effectively interacting with the active sites of antimicrobial protein receptors. This consistent finding supports that these new compounds’ practical and theoretical studies align regarding their antimicrobial activity.

Pharmaceuticals doi: 10.3390/ph17121631

Authors: Ioana Rada Popa Ilie Carmen Maximiliana Dobrea Anca Butuca Calin Homorodean Claudiu Morgovan Andreea Loredana Vonica-Tincu Felicia Gabriela Gligor Steliana Ghibu Adina Frum

Background/Objectives: Pasireotide (PAS) is a somatostatin receptor ligand (SRL) used to treat acromegaly, a chronic condition caused by excess growth hormone. While it offers significant benefits as a second-line treatment for uncontrolled acromegaly, its use raises major concerns due to hyperglycemic side effects and gastrointestinal issues, the latter being similar to those seen with first-generation SRLs. The aim of this study is to evaluate the real-world evidence on adverse drug reactions (ADRs) reported for PAS in the EudraVigilance database, in comparison to other established drug-based therapies for acromegaly. Methods: A descriptive analysis and a disproportionality analysis were conducted. Results: The fewest individual case safety reports (ICSRs) and adverse drug reactions (ADRs) were reported for PAS, with 698 (4%) ICSRs and 1,647 (4%) ADRs, which is even lower than for pegvisomant (PEG), which had 1765 (11%) ICSRs and 4842 (10%) ADRs. Both PAS and lanreotide (LAN) exhibited the lowest proportion of cases classified as serious. Among the total reported ADRs, those categorized as “Metabolic and nutrition disorders” were most frequent and severe for PAS (PAS—17.5% vs. OCT—4.6%, LAN—4.5%, and PEG—2.7%). Additionally, PAS demonstrated a higher likelihood of reporting endocrine disorders, which were frequently classified as serious, as well as stones affecting the hepatobiliary system compared to other drugs. Conclusions: Although PAS had the fewest ICSRs and ADRs, and less frequent serious ADRs, it had more reports frequently classified as serious in the “Metabolism and Nutrition Disorders” category (including events such as elevated blood glucose levels or diabetes) and “Endocrine Disorders” category compared to other SRLs and PEG. Furthermore, there was a higher likelihood of reporting hepatobiliary stones with PAS compared to OCT and PEG. This highlights the importance of adequately monitoring glycemic control and the biliary tract through ultrasound at the initiation and during follow-up of PAS therapy. Improved monitoring and reporting of these ADRs could enhance care for patients with acromegaly.

Pharmaceuticals doi: 10.3390/ph17121630

Authors: Dumitriţa Rugină Mihai Adrian Socaciu Madalina Nistor Zorita Diaconeasa Mihai Cenariu Flaviu Alexandru Tabaran Carmen Socaciu

Background/Objectives: Pentacyclic triterpenoids are increasingly studied as anticancer agents with many advantages compared to synthetic chemotherapeutics. The aim of this study was to prepare liposomal and nanostructured lipid formulations including a standardized extract of silver birch (Betula pendula) outer bark (TTs) and to evaluate their potential as anticancer agents in vitro, using Melanoma B16-F10 and Walker carcinoma cells. Methods: Appropriate solvents were selected for efficient TTs extraction, and original recipes were used to obtain Pegylated liposomes and nanolipid complexes with entrapped TTs, comparative to pure standards (betulinic acid and doxorubicin) in similar conditions. The composition, morphology, and sizes of all nanoformulations were checked by high-performance liquid chromatography/mass spectrometry, Transmission Electronic Microscopy, and Diffraction Light Scattering. The entrapment efficiency and its impact on cell viability, cell cycle arrest, and apoptosis by flow cytometry was also measured on both cancer cell lines. Conclusions: The standardized TTs, including betulin, lupeol, and betulinic acid, showed good stability and superior activity compared to pure betulinic acid. According to experimental data, TTs showed good entrapment in liposomal and NLC nanoformulations, both delivery systems including natural, biodegradable ingredients and enhanced bioavailability. The apoptosis and necrosis effects were more pronounced for TTs liposomal formulations in both types of cancer cells, with lower cytotoxicity compared to Doxorubicin, and can be correlated with their increased bioavailability.

Pharmaceuticals doi: 10.3390/ph17121629

Authors: Daniela Damiani Mario Tiribelli

Chimeric antigen receptor (CAR) T-cell therapy represents one of the most impressive advances in anticancer therapy of the last decade. While CAR T-cells are gaining ground in various B cell malignancies, their use in acute myeloid leukemia (AML) remains limited, and no CAR-T product has yet received approval for AML. The main limitation of CAR-T therapy in AML is the lack of specific antigens that are expressed in leukemic cells but not in their healthy counterparts, such as hematopoietic stem cells (HSCs), as their _targeting would result in an on-_target/off-tumor toxicity. Moreover, the heterogeneity of AML and the tendency of blasts to modify surface antigens’ expression in the course of the disease make identification of suitable _targets even more challenging. Lastly, AML’s immunosuppressive microenvironment dampens CAR-T therapeutic activities. In this review, we focus on the actual pitfalls of CAR T-cell therapy in AML, and we discuss promising approaches to overcome them.

Pharmaceuticals doi: 10.3390/ph17121627

Authors: Andrea Macejova Veronika Kovacova Ingrid Tonhajzerova Zuzana Visnovcova Nikola Ferencova Zuzana Mlyncekova Tomas Kukucka Igor Ondrejka

Background: Adolescent treatment resistant depression (TRD) is increasing in recent years. While ketamine showed rapid antidepressant effects in adult TRD studies, research on its effectiveness in adolescents is limited. Methods: This study examines the effects of intravenous ketamine vs. midazolam on depressive and anxiety symptomatology assessed by the Montgomery–Åsberg Depression Rating Scale (MADRS), Hamilton Anxiety Rating Scale (HAM-A), and Children’s Depression Inventory (CDI) at two time points—2 h after initial infusion (T0+2h) and 24 h after the end of the treatment (Te+24h) in a sample of 55 adolescent TRD females (27 receiving ketamine, 28 midazolam). Results: At T0+2h, within-group comparisons revealed a significant reduction in MADRS and HAM-A scores compared to baseline in the ketamine and midazolam groups. At Te+24h, both groups demonstrated similar significant reductions in MADRS, HAM-A, and CDI scores compared to baseline. The MADRS assessment in the ketamine group showed 33% and 59% responders, and in the midazolam group, 14% and 46% responders at T0+2h and Te+24h, respectively. HAM-A evaluation in the ketamine group revealed 33% and 56% responders, and in the midazolam group, 11% and 39% responders at T0+2h and at Te+24h, respectively. CDI rating discovered 11% and 44% responders in the ketamine group and 4% and 21% responders in the midazolam group at T0+2h and Te+24h, respectively. Moreover, inner tension significantly decreased in ketamine compared to the midazolam group at Te+24h. Conclusions: Ketamine showed a reduction in depressive and anxiety symptoms during a short-term period with particular efficacy in alleviating inner tension over midazolam, suggesting its potential advantages in specific symptom relief in rarely studied adolescent TRD.

Pharmaceuticals doi: 10.3390/ph17121628

Authors: Sara E. Maloney Norcross Leanna P. K. Levin Anthony J. Hickey David B. Hill

Natural and synthetic biopolymers are gaining popularity in the development of inhaled drug formulations. Their highly tunable properties and ability to sustain drug release allow for the incorporation of attributes not achieved in dry powder inhaler formulations composed only of micronized drugs, standard excipients, and/or carriers. There are multiple physiological barriers to the penetration of inhaled drugs to the epithelial surface, such as the periciliary layer mucus mesh, pulmonary macrophages, and inflammation and mucus compositional changes resulting from respiratory diseases. Biopolymers may facilitate transport to the epithelial surface despite such barriers. A variety of categories of biopolymers have been assessed for their potential in inhaled drug formulations throughout the research literature, ranging from natural biopolymers (e.g., chitosan, alginate, hyaluronic acid) to those synthesized in a laboratory setting (e.g., polycaprolactone, poly(lactic-co-glycolic acid)) with varying structures and compositions. To date, no biopolymers have been approved as a commercial dry powder inhaler product. However, advances may be possible in the treatment of respiratory diseases and infections upon further investigation and evaluation. Herein, this review will provide a thorough foundation of reported research utilizing biopolymers in dry powder inhaler formulations. Furthermore, insight and considerations for the future development of dry powder formulations will be proposed.

Pharmaceuticals doi: 10.3390/ph17121626

Authors: Ruiyi Qian Xue Zhao Dongbin Lyu Qingqing Xu Kai Yuan Xin Luo Wanying Wang Yang Wang Yutong Liu Yu Cheng Yingting Tan Fan Mou Chengmei Yuan Shunying Yu

Background: Restless legs syndrome (RLS) is a common sensorimotor sleep disorder that affects sleep quality of life. Much effort has been made to make progress in RLS pharmacotherapy; however, patients with RLS still report poor long-term symptom control. Methods: Comprehensive Mendelian randomization (MR) was performed to search for potential causal genes and drug _targets using the cis-pQTL and RLS GWAS data. Robustness was validated using the summary-based Mendelian randomization (SMR) method and co-localization analysis. Further evidence of pleiotropy of the _target genes and their potential side effects was provided by phenome-wide MR analysis (MR-PheWAS). Finally, molecular docking simulations were conducted on drug candidates corresponding to these _targets, which revealed promising binding affinities and interaction patterns and underscored the druggable potential of the _target gene. All of the analyses above were conducted in the context of Homo sapiens. Results: MAN1A2 showed a statistically significant result in the MR analysis, which was validated through SMR and co-localization analysis. The MR-PheWAS showed a low probability of pleiotropy and prospective side effects. Molecular docking was used to visualize the binding structure and fine affinity for MAN1A2 and the drugs predicted by DSigDB. Conclusions: Our study provides comprehensive evidence supporting MAN1A2 as a promising causal gene and therapeutic _target for RLS, offering insights into the underlying molecular mechanisms and paving the way for future drug development efforts.

Pharmaceuticals doi: 10.3390/ph17121624

Authors: Justin J. Odanga Sharon M. Anderson Sharon C. Presnell Edward L. LeCluyse Jingsong Chen Jessica R. Weaver

Background: Liver diseases are a global health concern. Many in vitro liver models utilize cryopreserved primary human hepatocytes (PHHs), which commonly undergo post-thaw processing through colloidal silica gradients to remove debris and enrich for a viable PHH population. Post-thaw processing effects on healthy PHHs are partially understood, but the consequences of applying disease-origin PHHs to post-thaw density gradient separation have not been described. Methods: Using the TruVivo® system, diseased, type 2 diabetes mellitus (T2DM), and fibrotic PHHs were cultured for 14 days after initially being subjected to either low-density (permissive) or high-density (selective) gradients using Percoll-based thawing medium. Results: Changes in functionality, including albumin and urea secretion and CYP3A4 activity, were measured in diseased, T2DM, and fibrotic PHHs enriched in low Percoll compared to PHHs enriched in high Percoll. Lipogenesis increased in the PHHs enriched in low Percoll. Higher expression of CK18 and TGF-β, two fibrotic markers, and changes in expression of the macrophage markers CD68 and CD163 were also measured. Conclusions: The use of Percoll for the enrichment of PHHs post-thaw results in differences in attachment and functionality, along with changes in diseased phenotypes, in the TruVivo® system.

Pharmaceuticals doi: 10.3390/ph17121625

Authors: María Carolina Otero Francisco Ceric Sebastián Miranda-Rojas Carolina Carreño Rachelly Escares María José Escobar Chiara Saracini Cristian Atala Ricardo Ramírez-Barrantes Felipe Gordillo-Fuenzalida

Hypericum perforatum, also known as St. John’s Wort, pericon, or yellow grass, is known for its antidepressant potential. It could represent a natural alternative to current pharmacological antidepressant treatments, which have a high incidence of side effects in patients and therefore lead to early dropouts. Through a bibliographic revision of clinical trials and information collected from scientific articles during the first period of 2020, we aimed to evaluate whether its administration could be beneficial in the treatment of mild-to-moderate depression, with fewer side effects compared to synthetic drugs. Among the main components, hypericin and hyperforin have been related to the observed antidepressant activity; therefore, their possible mechanism of action was reviewed and highlighted. Furthermore, patients receiving Hypericum extracts were less likely to withdraw from studies because of adverse effects compared to those receiving older standard antidepressants. This review aims to provide suggestions for an alternative treatment of mild-to-moderate depression disorder under the supervision of a medical doctor, since, although it appears to be a potentially efficient treatment with a low presence of adverse effects in comparison to synthetic antidepressants, it might also interact with other medications and lead to therapeutic failures if misused for self-medication.

Pharmaceuticals doi: 10.3390/ph17121623

Authors: Vitória Santório de São José Bruno Marques Vieira Vivaldo Moura Neto Lidia M. Lima

Background/Objectives: Glioblastoma is a malignant tumor with a poor prognosis for the patient due to its high lethality and limited chemotherapy available. Therefore, from the point of view of chemotherapy treatment, glioblastoma can be considered an unmet medical need. This has led to the investigation of new drugs for monotherapy or associations, acting by synergistic pharmacological mechanisms. Methods: Here, we propose the combination of Osimertinib (a potent EGFR inhibitor) and Gedatolisib (a potent PI3K/mTOR dual inhibitor) through an in vitro phenotypic study using five human GB lines and establish the cytotoxic potency, selectivity, and effect on proliferation, apoptosis, and cell cycle by simultaneously inhibiting EGFR, PI3K, and mTOR. Results: Cytotoxic potency of Gedatolisib and Osimertinib in the selected GB cell lines was determined, which highlighted the synergistic response from their combination and its impact on migration reduction, G0/G1 cell cycle arrest, GB cytotoxicity, and apoptosis-inducing effects for different GB cell lines. Conclusions: From the drug combination studies in phenotypic in vitro models, it was possible to suggest a new potential treatment for glioblastoma that justifies further safe in vivo phases of preclinical trials with the combination.

Pharmaceuticals doi: 10.3390/ph17121622

Authors: Cristiano da Rosa Larissa Kaori Maquedano Ian Lucas Bueno Fernando Augusto Lima Marson Giovanna Barbarini Longato

Background/Objectives: Treating chronic wounds incurs substantial costs for Brazil’s Unified Health System. Natural compounds, particularly propolis, are increasingly explored as low-cost alternatives due to their healing properties. Brazilian green propolis, distinct in its chemical composition, has garnered scientific interest. This study aimed to assess the healing effects of green propolis ointment on lower-limb ulcers from leprosy. Methods: A blinded, randomized clinical trial included 18 wounds in two groups: propolis ointment (G1) and control (G2), with evaluations conducted weekly for 61 days. Wound progress was monitored using morphometry and the Pressure Ulcer Scale for Healing (PUSH). Results: No participants exhibited sensitivity to the propolis. G1 showed significant initial healing: average wound area reduction (%) for G1 vs. G2 included 56.38 vs. 6.13–p < 0.001 (week 1); 79.51 vs. 24.16–p = 0.022 (week 4); and 84.33 vs. 39.73–p = 0.051 (week 7). In G1, the PUSH scores decreased from the beginning, whereas in G2, reductions were observed after three weeks. By week 5, 71.4% of G1 wounds scored below eight points, versus 33.3% in G2. G1 wounds exhibited a reduced area and exudate, as well as revitalized granulation tissue without adverse effects. Conclusions: The findings suggest that green propolis ointment is safe, supports tissue repair and may offer cost-effective treatment benefits. Standard wound dressings are selected to support all healing stages, with an emphasis on antimicrobial action, hemostasis to reduce exudate, and pain-reducing and non-irritant properties. Green propolis ointment meets these criteria, offering a cost-effective treatment that accelerates lesion reduction and encouraging leprosy patients to follow the therapeutic regimen.

Pharmaceuticals doi: 10.3390/ph17121621

Authors: Xiaoyan Liu Ruihu Du Tao Zhang Yingzi Li Ludi Li Zheng Yang Youbo Zhang Qi Wang

Background/Objectives: Spatholobi Caulis (SPC) is a medicinal plant that mainly grows in China and Southeast Asian countries and commonly used in clinics; the pharmacokinetic characteristics in humans need to be determined. This study was to establish the physiologically based pharmacokinetic (PBPK) models of multiple active constituents from SPC in rats, and predict the pharmacokinetic properties of rats with different dosages and extrapolated to humans. Methods: The parameters were collected based on our previous study and by prediction using ADMET Predictor software predict. The PBPK models for 3′-methoxydadizein (1), 8-O-methylretusin (2), daidzin (3), and isolariciresinol (4) administered orally to rats were established using GastroPlus software. These models were employed to simulate the pharmacokinetic properties in rats across various dosages, and subsequently extrapolated to humans. The calculated parameters including Cmax, Tmax, and AUC were compared with observed values. The accuracy of the PBPK models was assessed using fold-error (FE) values. Result: The FE values ranged from 1.03 to 1.52, meeting the PBPK model regulations where FE should be less than 2. The sensitivity analysis focusing on the absorption amount and AUC0→t of these four constituents in humans was also conducted. These results confirm the successful establishment of PBPK models of these four constituents from SPC in this study, and these models were applicable to predict pharmacokinetics across various doses and extrapolate across species. Conclusions: The PBPK models of four constituents can be used to predict the pharmacokinetic characteristics in humans after oral administration of SPC and provide useful data for safe and rational medication in clinical practice.

Pharmaceuticals doi: 10.3390/ph17121620

Authors: Béla E. Tóth István Takács Kristóf Kádár Sara Mirani Miklós Vecsernyés Péter Lakatos

Background/Objectives: Epidemiological data on vitamin D status revealed that, despite various dosage and durations of supplementation, the effectiveness often fails to achieve optimal outcomes. The need for higher doses than previously recommended was suggested, but several modifying factors should be considered, including the level of deficiency, and BMI. The objectives of this post hoc evaluation are to characterize treatment effectiveness based on the applied dose, duration and BMI; and to assess the safety aspects associated with rapid repletion of vitamin D. Methods: Vitamin D deficient subjects selected in the post-hoc analysis: seventy patients included from a combined loading-maintenance supplementation (300,000 IU followed by 60,000 IU) protocol and 62 deficient subjects who received a low-dose maintenance (1000 IU/day) therapy. The risk of overload and the incidence of hypercalciuria and hypercalcemia resulting from loading or post-loading maintenance were investigated. Results: The moderate–fast-loading schedule of 60,000 IU per week for 5 weeks, effectively achieves the _target in 25(OH)D levels over 30 ng/mL for all deficient subjects, regardless of their BMI. Slower loading with lower weekly doses confirms the safety of supplementation, but the effectiveness is dependent on the subjects’ BMI; overweight and obese patients require higher doses to reach the same vitamin D levels. No difference in safety parameters observed compared to low-dose therapies. Conclusions: The loading treatment involving a total dose of 300,000 IU administered over 5 or 10 weeks is effective for repletion, does not lead to 25(OH)D overload, and poses no additional risks of hypercalcemia or hypercalciuria. Furthermore, there are no safety concerns regarding changes in bone resorption markers. A combination of the loading treatment with a subsequent maintenance dose of 2000 IU daily is adequate to achieve the _target vitamin D levels.

Pharmaceuticals doi: 10.3390/ph17121619

Authors: Geovanna Nallely Quiñonez-Bastidas Lucia Elhy Grijalva-Contreras Selene Isabel Patiño-Camacho Andrés Navarrete

Trigeminal neuralgia (TN) is chronic pain caused by damage to the somatosensorial system on the trigeminal nerve or its branches, which involves peripheral and central dysfunction pain pathways. Trigeminal pain triggers disruptive pain in regions of the face, including within and around the mouth. Besides clinical experiences, translating the language of suffering into scientific terminology presents substantial challenges. Due to the complex and multifactorial pathophysiology underlying trigeminal pain, elucidating its social impact presents significant difficulties. Carbamazepine and oxcarbazepine are first-line treatments for TN, achieving approximately 50% pain reduction in 60–70% of treated patients. However, their efficacy is often limited by common side effects, such as dizziness, vertigo, nausea, seizures, and cognitive symptoms. In some cases, patients experience severe side effects, including myelosuppression, hyponatremia, hormonal imbalances, liver toxicity, suicidal ideation, teratogenicity, and other adverse reactions. Given these clinical limitations, the search for new painkiller candidates continues. Hence, we focused this review on salvinorin A (SalA), a natural agonist of κ-opioid receptors (KORs), which demonstrated anti-nociceptive, anti-inflammatory, and anti-neuropathic properties in various experimental models of the spinal sensory system. Furthermore, preclinical evidence indicates that SalA does not induce dependence and demonstrates a favorable toxicological and safety profile in comparison with currently marketed opioid drugs. We propose Salvinorin A as a promising candidate for treating trigeminal neuralgia, offering the potential for reduced adverse effects.

Pharmaceuticals doi: 10.3390/ph17121618

Authors: Stefano Comai Sara De Martin Andrea Mattarei Clotilde Guidetti Marco Pappagallo Franco Folli Andrea Alimonti Paolo L. Manfredi

Uncompetitive NMDAR (N-methyl-D-aspartate receptor) antagonists restore impaired neural plasticity, reverse depressive-like behavior in animal models, and relieve major depressive disorder (MDD) in humans. This review integrates recent findings from in silico, in vitro, in vivo, and human studies of uncompetitive NMDAR antagonists into the extensive body of knowledge on NMDARs and neural plasticity. Uncompetitive NMDAR antagonists are activity-dependent channel blockers that preferentially _target hyperactive GluN2D subtypes because these subtypes are most sensitive to activation by low concentrations of extracellular glutamate and are more likely activated by certain pathological agonists and allosteric modulators. Hyperactivity of GluN2D subtypes in specific neural circuits may underlie the pathophysiology of MDD. We hypothesize that neural plasticity is epigenetically regulated by precise Ca2+ quanta entering cells via NMDARs. Stimuli reach receptor cells (specialized cells that detect specific types of stimuli and convert them into electrical signals) and change their membrane potential, regulating glutamate release in the synaptic cleft. Free glutamate binds ionotropic glutamatergic receptors regulating NMDAR-mediated Ca2+ influx. Quanta of Ca2+ via NMDARs activate enzymatic pathways, epigenetically regulating synaptic protein homeostasis and synaptic receptor expression; thereby, Ca2+ quanta via NMDARs control the balance between long-term potentiation and long-term depression. This NMDAR Ca2+ quantal hypothesis for the epigenetic code of neural plasticity integrates recent psychopharmacology findings into established physiological and pathological mechanisms of brain function.

Pharmaceuticals doi: 10.3390/ph17121617

Authors: Ahmed M. Hassan Hattan S. Gattan Arwa A. Faizo Mohammed H. Alruhaili Azzah S. Alharbi Leena H. Bajrai Ibrahim A. AL-Zahrani Vivek Dhar Dwivedi Esam I. Azhar

Background/Objectives: Monkeypox is a re-emerging viral disease with features of infectiously transmitted zoonoses. It is now considered a public health priority because of its rising incidence and transmission from person to person. Monkeypox virus (MPXV) VP39 protein is identified as an essential protein for replication of the virus, and therefore, it is a potential _target for antiviral drugs. Methods: This work analyzes the binding affinities and the differential conformational stability of three _target compounds and one control compound with the VP39 protein through multiple computational methods. Results: The re-docking analysis revealed that the compounds had high binding affinities towards the _target protein; among these compounds, compounds 1 and 2 showed the highest binding energies in the virtual screening, and thus, these were considered as the most active inhibitor candidates. Intermolecular interaction analysis revealed distinct binding mechanisms. While compound 1 had very strong hydrogen bonds and hydrophobic interactions, compound 2 had numerous water-mediated interactions, and compound 3 had only ionic and hydrophobic contacts. In molecular dynamic simulations, compounds 1 and 2 showed that the protein–ligand complexes had a stable conformation, with protein RMSD values around 2 Å for both compounds. In contrast, compound 3 was slightly flexible, and the control compound was more flexible. MM/GBSA analysis again supported these results, which gave the binding free energies that were also supportive for these compounds. Conclusions: Notably, all the selected compounds, especially compounds 1 and 2, demonstrate high binding affinity. Therefore, these compounds can be further tested as antiviral agents against monkeypox treatment.

Pharmaceuticals doi: 10.3390/ph17121616

Authors: Boris Parra Maximiliano Sandoval Vicente Arriagada Luis Amsteins Cristobal Aguayo Andrés Opazo-Capurro Arnaud Dechesne Gerardo González-Rocha

Background/Objectives: Antimicrobial resistance (AMR) is a major public health threat, which is exacerbated by the lack of new antibiotics and the emergence of multidrug-resistant (MDR) superbugs. Comprehensive efforts and alternative strategies to combat AMR are urgently needed to prevent social, medical, and economic consequences. Pseudomonas aeruginosa is a pathogen responsible for a wide range of infections, from soft tissue infections to life-threatening conditions such as bacteremia and pneumonia. Bacteriophages have been considered as a potential therapeutic option to treat bacterial infections. Our aim was to isolate phages able to infect MDR P. aeruginosa strains. Methods: We isolated two lytic phages, using the conventional double layer agar technique (DLA), from samples obtained from the influent of a wastewater treatment plant in Concepción, Chile. The phages, designated as PaCCP1 and PaCCP2, were observed by electron microscopy and their host range was determined against multiple P. aeruginosa strains using DLA. Moreover, their genomes were sequenced and analyzed. Results: Phage PaCCP1 is a member of the Septimatrevirus genus and phage PaCCP2 is a member of the Pbunavirus genus. Both phages are tailed and contain dsDNA. The genome of PaCCP1 is 43,176 bp in length with a GC content of 54.4%, encoding 59 ORFs, one of them being a tRNA gene. The genome of PaCCP2 is 66,333 bp in length with a GC content of 55.6%, encoding 102 non-tRNA ORFs. PaCCP1 is capable of infecting five strains of P. aeruginosa, whereas phage PaCCP2 is capable of infecting three strains of P. aeruginosa. Both phages do not contain bacterial virulence or AMR genes and contain three and six putative Anti-CRISPR proteins. Conclusions: Phages PaCCP1 and PaCCP2 show promise as effective treatments for MDR P. aeruginosa strains, offering a potential strategy for controlling this clinically important pathogen through phage therapy.

Pharmaceuticals doi: 10.3390/ph17121615

Authors: Madumani Amararathna David W. Hoskin Kerry B. Goralski H. P. Vasantha Rupasinghe

Oral supplementation of anthocyanins-rich haskap (Lonicera caerulea) berry (HB) reduces 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung tumorigenesis, cytotoxicity, DNA damage, and modulated inflammation in vitro and in vivo. The procarcinogen NNK is metabolically activated by cytochrome P450 (P450) enzymes, producing reactive metabolites that induce lung carcinogenesis. Hypothesis: Therefore, we hypothesized that the HB-modulated protective effect against NNK could be due to its ability to suppress P450 enzymes. Methods: HB (6 mg of cyanidin-3-O-glucoside [C3G] in 0.2 g of HB/mouse/day) was given to A/J mice as a dietary supplement following subsequent administration of NNK (100 mg/kg body weight). The liver tissues of mice were analyzed to determine the expression of P450s and metabolites. Results: HB upregulated the expression of cyp2a4 and cyp2a5 mRNA and nuclear receptor/transcription factor (PPARα) in NNK-deprived hepatic tissues. With NNK, HB downregulated the expression of cyp2a4 and cyp2a5 and facilitated the formation of non-carcinogenic NNK metabolites. Molecular docking indicated a high binding affinity and strong hydrophobic interactions between C3G and its major metabolites, peonidin-3-O-glucoside, petunidin-3-O-glucoside, peonidin and cyanidin with Cyp2a5 and with human P450 homologue CYP2A13. Conclusions: HB could be a potential dietary supplement to inhibit the P450 activated NNK carcinogenic metabolites formation. Hence, inhibiting the activation of NNK by lung CYP2A13 through dietary HB supplementation could be a strategy to reduce lung carcinogenesis among smokers. Understanding the effect of HB on the activity of CYP2A13 in human studies is necessary before recommending these natural compounds as therapeutics.

Pharmaceuticals doi: 10.3390/ph17121614

Authors: Hend I. Almohamady Yasmin Mortagi Shadeed Gad Sawsan Zaitone Reem Alshaman Abdullah Alattar Fawaz E. Alanazi Pierre A. Hanna

Background/Objectives: Drugs exhibiting poor aqueous solubility present a challenge to efficient delivery to the site of action. Spanlastics (a nano, surfactant-based drug delivery system) have emerged as a powerful tool to improve solubility, bioavailability, and delivery to the site of action. This study aimed to better understand factors affecting the physicochemical properties of spanlastics, quantify their effects, and use them to enhance the bioavailability of famotidine (FMT), a model histamine H2 receptor antagonist (BCS class IV). Methods: FMT was incorporated into nano-spanlastics drug delivery system. The ethanol injection method, Box–Behnken design, and mathematical modeling were utilized to fabricate famotidine-loaded nano-spanlastics and optimize the formula. Spanlastics were characterized for their particle size, polydispersity index, zeta potential, entrapment efficiency, drug loading, compatibility of the excipients (using DSC), in vitro drug release, and in vivo pharmacokinetics. Results: Span 60 (the non-ionic surfactant) and tween 60 (the edge activator) gave rise to spanlastics with the best characteristics. The optimal spanlastic formulation exhibited small particle size (<200 nm), appropriate polydispersity index (<0.4), and zeta potential (>−30 mV). The entrapment efficiency and drug loading of the optimum formula assured its suitability for hydrophobic drug entrapment as well as practicability for use. DSC assured the compatibility of all formulation components. The drug release manifested a biphasic release pattern, resulting in a fast onset and sustained effect. Spanlastics also showed enhanced Cmax, AUC0–24, and bioavailability. Conclusions: Spanlastics manifested improved FMT dissolution, drug release characteristics, membrane permeation, and pharmacokinetic behavior.

Pharmaceuticals doi: 10.3390/ph17121612

Authors: Sindy Magri Roque Ana Carolina Furian Marcela Kim Takemoto Marta Cristina Teixeira Duarte Rafaela Durrer Parolina Adriano Luís Roque Nelson Duran Janaína de Cássia Orlandi Sardi Renata Maria Teixeira Duarte Karina Cogo Muller

Introduction: Simvastatin is an antilipidemic drug that has already demonstrated antibacterial activities on oral and non-oral microorganisms. Silver nanoparticles also exhibit antimicrobial properties, particularly for coating implant surfaces. In this study, we evaluated the effects of combining simvastatin with silver nanoparticles on the formation and viability of biofilms consolidated on titanium discs. Methods: Silver nanoparticles were first biosynthesized using the fungus Fusarium oxysporum and then characterized using Dynamic Light Scattering, X-ray Diffraction, Transmission Electron Microscopy, and energy dispersive spectroscopy. Species of Streptococcus oralis, Streptococcus mutans, Porphyromonas gingivalis, Methicillin-sensitive Staphylococcus aureus, and Methicillin-resistant Staphylococcus aureus were used and tested using Minimum Inhibitory Concentration assays with concentrations of silver nanoparticles and simvastatin alone and in combination. Biofilm inhibition and viability tests were performed on titanium surfaces. Toxicity tests were also performed on Galleria mellonella moth larvae. Results: The silver nanoparticles had a spherical shape without the formation of aggregates as confirmed by Transmission Electron Microscopy. Dynamic Light Scattering revealed nanoparticles with an average diameter of 53.8 nm (±1.23 nm), a polydispersity index of 0.23 and a zeta potential of −25 mV (±2.19 mV). The silver nanoparticles inhibited the growth of the strains tested in the range of 0.001592 and 63.75, while simvastatin alone inhibited the growth of the same strains in the range of 3.125–62.5 µg/mL. The antibacterial activity test of the combination of the two substances showed a reduction in the Minimum Inhibitory Concentration of about two to eight times, showing synergistic effects on Staphylococcus aureus and additive effects on Streptococcus oralis and Porphyromonas gingivalis. As for biofilm, sub-inhibitory concentrations of the combination of substances showed better antibacterial activity in inhibiting the formation of Streptococcus oralis biofilm, and this combination also proved effective in eradicating already established biofilms compared to the substances alone. The combination of silver nanoparticles and simvastatin showed low toxicity to Galleria mellonella moth larvae. Conclusions: The results presented indicate that the combination of the two substances could be an alternative for the prevention and reduction of biofilms on implants. These findings open up new possibilities in the search for alternatives for the treatment of peri-implant infections, as well as the possibility of using lower doses compared to single drugs, achieving the same results and reducing potential toxic effects.

Pharmaceuticals doi: 10.3390/ph17121613

Authors: Gayane A. Atazhanova Yana K. Levaya Karakoz Zh. Badekova Margarita Yu. Ishmuratova Marlen K. Smagulov Zhanna O. Ospanova Elina M. Smagulova

This review is devoted to a systematic analysis of studies aimed at investigating plant extracts, essential oils and phytochemical compounds capable of inhibiting Streptococcus mutans biofilm formation. This paper investigates the effect of extracts, essential oils and individual plant compounds on inhibiting the biofilm formation of Streptococcus mutans, one of the major pathogens responsible for the development of dental caries. Using cultural microbiology and molecular biology techniques, the authors describe the mechanisms by which plant samples reduce Streptococcus mutans adhesion and growth. The results show that several plant components have antibacterial properties, contributing to the reduction of Streptococcus mutans colony numbers and inhibiting the synthesis of extract-exopolysaccharide matrices required for biofilm formation. This work highlights the potential of botanicals in inhibiting Streptococcus mutans biofilm formation, which can be applied as natural antimicrobial agents in the prevention and treatment of dental diseases. Views on the use of these plant extracts and their components in dental preparations such as toothpastes, rinses and gels aimed at preventing dental caries are evaluated. The review shows the relevance of the research to optimizing the use of plant extracts, essential oils, individual compounds and their active actions in the control of Streptococcus mutans biofilms.

Pharmaceuticals doi: 10.3390/ph17121611

Authors: Alin Dumitru Ciubotaru Carmen-Ecaterina Leferman Bogdan-Emilian Ignat Anton Knieling Delia Lidia Salaru Dana Mihaela Turliuc Liliana Georgeta Foia Lorena Dima Bogdan Minea Luminita Diana Hritcu Bogdan Ionel Cioroiu Laura Stoica Ioan-Adrian Ciureanu Alin Stelian Ciobica Bogdan Alexandru Stoica Cristina Mihaela Ghiciuc

Background/Objectives: Ongoing challenges in epilepsy therapy warrant research on alternative treatments that offer improved efficacy and reduced side effects. Designed to enhance mitochondrial _targeting and increase bioavailability, mitocurcumin (MitoCur) was evaluated for the first time as an antiepileptic agent, with curcumin (Cur) and sodium valproate (VPA), a standard antiepileptic drug, included for comparison. This study investigated the effects on seizure onset, severity, and progression in a zebrafish model of pentylenetetrazole (PTZ)-induced seizures and measured the concentrations of the compounds in brain tissue. Methods: Zebrafish were pre-treated with MitoCur and Cur (both at 0.25 and 0.5 µM doses) and VPA (0.25 and 0.5 mM) and observed for four minutes to establish baseline locomotor behavior. Subsequently, the animals were exposed to a 5 mM PTZ solution for 10 min, during which seizure progression was observed and scored as follows: 1—increased swimming; 2—burst swimming, left and right movements; 3—circular movements; 4—clonic seizure-like behavior; 5—loss of body posture. The studied compounds were quantified in brain tissue through HPLC and LC-MS. Results: Compared to the control group, all treatments reduced the distance moved and the average velocity, without significant differences between compounds or doses. During PTZ exposure, seizure latencies revealed that all treatments effectively delayed seizure onset up to score 4, demonstrating efficacy in managing moderate seizure activity. Notably, MitoCur also provided significant protection against the most severe seizure score (score 5). Brain tissue uptake analysis indicated that MitoCur achieved higher concentrations in the brain compared to Cur, at both doses. Conclusions: These results highlight the potential of MitoCur as a candidate for seizure management.

Pharmaceuticals doi: 10.3390/ph17121610

Authors: Sophia Athanasopoulou Eleni Spanidi Eleni Panagiotidou Andrea Cavagnino Anaïs Bobier Konstantinos Gardikis

Background/Objectives: Vine leaves are a bulky by-product that are disposed of and treated as waste in the wine production process. In the present study polyphenols from vine leaves were extracted and simultaneously encapsulated in a new delivery system consisting of liposomes and cyclodextrins. This system was further combined with propolis polyphenols encapsulated in cyclodextrins, resulting in a colloidal suspension for the release of antioxidants in a time-controlled way, the rate of which depends on the ratio of the materials. The result is a raw material that exhibits antioxidant and ECM protective effects when administered in skin fibroblasts (NHDFs). Methods: The antioxidant and ECM promoting efficacy of the produced raw material was assessed by the Folin–Ciocalteu method, DPPH assay, and in cellulo assays in fibroblasts, such as the cell viability assay, scratch assay, cell migration assay, gene expression analysis, and immunofluorescence analysis, for the detection, visualization, and quantification of collagen-I, collagen-IIIa, and elastin signals and collagenase assay. Results: Treatment of NHDFs with the combinatorial delivery system promoted collagen and elastin synthesis and deposition in normal conditions and, upon induced external stress, as assessed by in vitro transcriptomic and proteomic analysis. A significant inhibition of collagenase was also observed, suggesting a multi_targeted efficacy of the active ingredients also by preventing collagen degradation. Conclusions: Therefore, this liposome–cyclodextrin encapsulated polyphenol complex represents a novel bioactive ingredient with promising skin applications.

Pharmaceuticals doi: 10.3390/ph17121609

Authors: Gary Punjabi Elena Ramírez

Background/Objectives: This study evaluated the appropriateness of transmucosal immediate-release fentanyl (TIRF) prescriptions in a Madrid emergency room during 2019 and 2022, following a 2018 warning about off-label use. Methods: TIRF prescription in the emergency room search yielded 993 patients in 2019 and 1499 in 2022, of which 140 were randomized for the study, 70 in 2019, and 70 in 2022. Dose appropriateness and indication for TIRF were analyzed according to established criteria. Results: Despite a high prevalence of cancer diagnoses (77.9%, 109/140), only 32.9% (46/140) of patients met the appropriateness criteria pre-hospitalization. This improved to 42.5% (51/120) at discharge, but the change was not statistically significant overall. However, focusing on surviving patients reveals a significant improvement in appropriateness, increasing from 30.83% (37/120) to 42.50% (p = 0.002). This improvement was particularly pronounced in 2022 (p = 0.0269), but not in 2019 (p = 0.0771). Interestingly, appropriateness in patients with prior TIRF prescriptions remained relatively stable from pre-hospitalization (46.75%) to discharge (48.78%). A concerningly high proportion of patients with cancer diagnoses (68.75%) received low-dose opioid therapy (<60 MME) at discharge, and 36.8% of patients over 80 years old were co-prescribed benzodiazepines, contradicting prescribing guidelines. Conclusions: This study found inappropriate TIRF prescriptions were common in an emergency room setting, often due to low pre-hospital opioid doses. While hospitalization improved TIRF appropriateness in survivors, especially in 2022, concerning prescribing practices persisted. This emphasizes the need for better education and interventions to ensure safe and effective TIRF use.

Pharmaceuticals doi: 10.3390/ph17121608

Authors: Sabrin Haddad Manuel Hessenberger Cornelia Ablinger Clarissa Eibl Ruslan Stanika Marta Campiglio Gerald J. Obermair

Background: α2δ proteins regulate membrane trafficking and biophysical properties of voltage-gated calcium channels. Moreover, they modulate axonal wiring, synapse formation, and trans-synaptic signaling. Several rare missense variants in CACNA2D1 (coding for α2δ-1) and CACNA2D3 (coding for α2δ-3) genes were identified in patients with autism spectrum disorder (ASD). However, the pathogenicity of these variants is not known, and the molecular mechanism by which α2δ proteins may contribute to the pathophysiology of autism is, as of today, not understood. Therefore, in this study we functionally characterized two heterozygous missense variants in α2δ-1 (p.R351T) and α2δ-3 (p.A275T), previously identified in patients with ASD. Methods: Electrophysiological recordings in transfected tsA201 cells were used to study specific channel-dependent functions of mutated α2δ proteins. Membrane expression, presynaptic _targeting, and trans-synaptic signaling of mutated α2δ proteins were studied upon expression in murine cultured hippocampal neurons. Results: Homologous expression of both mutated α2δ proteins revealed a strongly reduced membrane expression and synaptic localization compared to the corresponding wild type α2δ proteins. Moreover, the A275T mutation in α2δ-3 resulted in an altered glycosylation pattern upon heterologous expression. However, neither of the mutations compromised the biophysical properties of postsynaptic L-type (CaV1.2 and CaV1.3) and presynaptic P/Q-type (CaV2.1) channels when co-expressed in tsA201 cells. Furthermore, presynaptic expression of p.R351T in the α2δ-1 splice variant lacking exon 23 did not affect trans-synaptic signaling to postsynaptic GABAA receptors. Conclusions: Our data provide evidence that the pathophysiological mechanisms of ASD-causing mutations of α2δ proteins may not involve their classical channel-dependent and trans-synaptic functions. Alternatively, these mutations may induce subtle changes in synapse formation or neuronal network function, highlighting the need for future α2δ protein-linked disease models.

Pharmaceuticals doi: 10.3390/ph17121607

Authors: Stephanie Perrino Udi Vazana Ofer Prager Lior Schori Gal Ben-Arie Anna Minarik Yinhsuan Michely Chen Orçun Haçariz Masakazu Hashimoto Yiftach Roth Gabriel S. Pell Alon Friedman Pnina Brodt

Background: Glioblastoma multiforme is an aggressive malignancy with a dismal 5-year survival rate of 5–10%. Current therapeutic options are limited, due in part to drug exclusion by the blood–brain barrier (BBB). We have previously shown that high-amplitude repetitive transcranial magnetic stimulation (rTMS) in rats allowed the delivery across the BBB of an IGF signaling inhibitor—IGF-Trap. The objective of this study was to assess the therapeutic effect of IGF-Trap when delivered in conjunction with rTMS on the intracerebral growth of glioma. Results: We found that systemic administration of IGF-Trap without rTMS had a minimal effect on the growth of orthotopically injected glioma cells in rats and mice, compared to control animals injected with vehicle only or treated with sham rTMS. In rats treated with a combination of rTMS and IGF-Trap, we observed a growth retardation of C6 tumors for up to 14 days post-tumor cell injection, although tumors eventually progressed. In mice, tumors were detectable in all control groups by 14–17 days post-injection of glioma GL261 cells and progressed rapidly thereafter. In mice treated with rTMS prior to IGF-Trap administration, tumor growth was inhibited or delayed, although the tumors also eventually progressed. Conclusion: The results showed that rTMS could increase the anti-tumor effect of IGF-Trap during the early phases of tumor growth. Further optimization of the rTMS protocol is required to improve survival outcomes.

Pharmaceuticals doi: 10.3390/ph17121604

Authors: Saikat Fakir Nektarios Barabutis

Background/Objectives: Endothelial hyperpermeability is the hallmark of severe disease, including sepsis and acute respiratory syndrome (ARDS). The development of medical countermeasures to treat the corresponding illness is of utmost importance. Synthetic somatostatin analogs (SSA) are FDA-approved drugs prescribed in patients with neuroendocrine tumors, and they act via growth hormone (GH) suppression. Preclinical investigations suggest that Octreotide (OCT) alleviates Lipopolysaccharide (LPS)-induced injury. The aim of the study is to investigate the involvement of activating transcription factor 6 (ATF6) in the protective effects of OCT in endothelial dysfunction. To the best of our knowledge, the available information on that topic is limited. Methods: Human lung microvascular endothelial cells (HULEC-5a) and bovine pulmonary artery endothelial cells (BPAEC) which expressed elevated levels of ATF6 due to AA147 were exposed to OCT or vehicle. Protein expression, endothelial permeability, and reactive oxygen species (ROS) generation were assessed utilizing Western blot analysis, Fluorescein isothiocyanate (FITC)-Dextran assay, and Dichlorofluorescein diacetate measurements, respectively. Results: Our observations suggest that ATF6 activation significantly improves OCT-induced endothelial barrier enhancement. This combination led to increased expression of binding immunoglobulin protein (BiP) and glucose-regulated protein 94 (Grp94), which are downstream unfolded protein response (UPR) _targets. Moreover, ATF6 activation prior to OCT treatment resulted in decreased activation of myosin light chain 2 (MLC2) and cofilin; and reduced reactive oxygen species (ROS) generation. ATF6 activation enhanced the anti-inflammatory effects of OCT, as reflected in the suppression of transducer and activator of transcription (STAT) 1, STAT3, and P38 phosphorylation. Conclusions: Our findings suggest that ATF6 activation prior to OCT treatment enhances the beneficial effects of OCT in the endothelium.

Pharmaceuticals doi: 10.3390/ph17121605

Authors: Heba M. Aboud Adel A. Ali Nada H. Mohammed Ahmed H. E. Hassan Eun Joo Roh Shahira F. El Menshawe

Background/objectives: Idiopathic pulmonary fibrosis (IPF) is a prevalent interstitial lung disease that typically progresses gradually, leading to respiratory failure and ultimately death. IPF can be treated with the tyrosine kinase inhibitor, nintedanib (NTD), owing to its anti-fibrotic properties, which ameliorate the impairment of lung function. This study aimed to formulate, optimize, and assess NTD-loaded ufasomes (NTD-UFSs) as a nanosystem for its pulmonary _targeting to snowball the bioavailability and therapeutic efficacy of the drug. Methods: To investigate the influence of numerous factors on NTD-UFSs assembly and to determine the optimal formulation, Box–Behnken statistical design was implemented with the assistance of Design-Expert® software. The thin-film hydration strategy was employed to fabricate NTD-UFSs. The optimum NTD-UFSs formulation was subsequently selected and subjected to additional evaluations. Also, using a rat model, a comparative pharmacokinetic analysis was scrutinized. Results: The optimal NTD-UFSs elicited an accumulative release of 65.57% after 24 h, an encapsulation efficiency of 62.51%, a zeta potential of −36.07 mV, and a vesicular size of 364.62 nm. In addition, it disclosed remarkable stability and a continuous cumulative release pattern. In vivo histopathological studies ascertained the tolerability of NTD-UFSs administered intratracheally. According to the pharmacokinetic studies, intratracheal NTD-UFSs administration manifested a significantly higher AUC0–∞ value than oral and intratracheal NTD suspensions, by approximately 5.66- and 3.53-fold, respectively. Conclusions: The findings of this study proposed that UFSs might be a promising nanoparadigm for the non-invasive pulmonary delivery of NTD.

Pharmaceuticals doi: 10.3390/ph17121606

Authors: Veysel Toprak İlhan Özdemir Şamil Öztürk Orhan Yanar Yusuf Ziya Kizildemir Mehmet Cudi Tuncer

Background/Objectives: Ovarian cancer has the highest mortality rate in the world. Treatment methods are listed as surgery, chemotherapy, and radiotherapy, depending on the stage of cancer, but developing resistance to chemotherapy increases the need for alternative agents that act on the same pathways. The effects of rosmarinic acid (RA) and doxorubicin (DX) on the activation of FOXP3, an important tumor suppressor gene, in OVCAR3 cells were examined. Materials and Methods: In this study, a human ovarian adenocarcinoma cell line was used. MTT analysis was performed to reveal the result of RA and DX on ovarian cancer cell proliferation. Expression levels of FOXP3 for cell proliferation and Capase-3 for apoptosis were determined by RT-qPCR. The wound healing model was applied to determine cell migration rates. The results were evaluated with one-way ANOVA in an SPSS 20.0 program as p ≤ 0.05. Results: It was determined that RA and DX alone and in combination inhibited the proliferation of OVCAR3 cells in different doses for 24, 48, and 72 h, and caused the cells to die by causing them to undergo apoptosis. Caspase-3 expression increased approximately tenfold in OVCAR3 cells, while FOXP3 expression was upregulated only in RA treatment and was downregulated in DX and RA + DX treatments. Conclusions: According to the results of our study, it was determined that the FOXP3 signaling pathway related to apoptosis, and proliferation was affected by the combination treatment of RA and DX in the OVCAR3 cancer cell line. This shows that RA will gain an important place in cancer treatment with more comprehensive study.

Pharmaceuticals doi: 10.3390/ph17121602

Authors: Ahmad Elbahnsi Balint Dudas Isabelle Callebaut Alexandre Hinzpeter Maria A. Miteva

The ATP-binding cassette (ABC) and solute carrier (SLC) transporters play pivotal roles in cellular transport mechanisms, influencing a wide range of physiological processes and impacting various medical conditions. Recent advancements in structural biology and computational modeling have provided significant insights into their function and regulation. This review provides an overview of the current knowledge of human ABC and SLC transporters, emphasizing their structural and functional relationships, transport mechanisms, and the contribution of computational approaches to their understanding. Current challenges and promising future research and methodological directions are also discussed.

Pharmaceuticals doi: 10.3390/ph17121603

Authors: Cristina Sempio Jorge Campos-Palomino Jelena Klawitter Erica N. Peters Laura MacNair Mehdi Haghdoost Marcel O. Bonn-Miller Amy Harrison Shanna Babalonis Uwe Christians Jost Klawitter

Background: Tetrahydrocannabivarin (THCV) is a phytocannabinoid commonly found in cannabis with potential pharmacological properties; however, its post-acute pharmacokinetics (PK) in humans have not been studied yet. THCV has two isomers, Δ9- and Δ8-THCV, which seem to have different pharmacological properties. We investigated the PK of the Δ8-THCV isomer after oral administration as part of a two-phase, dose-ranging, placebo-controlled trial in healthy participants. Methods: Participants (n = 21) were enrolled in six study sessions and randomly received the following doses of a medium-chain triglyceride (MCT) oil oral formulation of Δ8-THCV: placebo, 12.5 mg, 25 mg, 50 mg, 100 mg, and 200 mg. Plasma samples from 15 participants were collected up to 8 h after administration and were analyzed by a validated two-dimensional high-performance liquid chromatography–tandem mass spectrometry assay. The trial was registered on clinicaltrials.gov (NCT05210634). Results: After oral administration, 11-nor-9-carboxy-Δ8-THCV (Δ8-THCV-COOH) was the main metabolite detected. The median time-to-maximum concentration (tmax) ranged 3.8–5.0 h across doses for Δ8-THCV and 4.6–5.3 h for Δ8-THCV-COOH. The maximum concentration (Cmax) and area under the concentration–time curve over the observation period (AUClast) appeared to be dose-linear. Median AUClast increased 2.3- to 4.8-fold and 1.7- to 2.9-fold for Δ8-THCV and Δ8-THCV-COOH, respectively, every two-fold increase in the dose. The isomers Δ9-THCV and Δ9-THCV-COOH were detected in plasma, despite being undetected in the formulated drug product analyzed by a third-party laboratory. Conclusions: For the first time, we report the pharmacokinetics of Δ8-THCV and its major metabolites after oral administration in humans. Δ8-THCV AUClast showed dose linearity but the observed possible conversion to the Δ9-THCV isomer should be further studied.

Pharmaceuticals doi: 10.3390/ph17121601

Authors: Chris-Tiann Roberts Nicole Raabe Lara Wiegand Ashraf Kadar Shahib Mojgan Rastegar

Metformin is a commonly used drug for treating type 2 diabetes. Metformin is an inexpensive drug with low/no side effects and is well tolerated in human patients of different ages. Recent therapeutic strategies for human disease have considered the benefits of drug repurposing. This includes the use of the anti-diabetic drug metformin. Accordingly, the anti-inflammatory, anti-cancer, anti-viral, neuroprotective, and cardioprotective potentials of metformin have deemed it a suitable candidate for treating a plethora of human diseases. As results from preclinical studies using cellular and animal model systems appear promising, clinical trials with metformin in the context of non-diabetes-related illnesses have been started. Here, we aim to provide a comprehensive overview of the therapeutic potential of metformin in different animal models of human disease and its suggested relationship to epigenetics and ailments with epigenetic components.

Pharmaceuticals doi: 10.3390/ph17121600

Authors: Reem Binsuwaidan Thanaa A. El-Masry Maysa M. F. El-Nagar Enas I. El Zahaby Mohamed M. S. Gaballa Maisra M. El-Bouseary

Background: The potent antioxidant lycopene has attracted a large amount of research attention given its potential health benefits. We aimed to assess the antimicrobial, anti-inflammatory, and antioxidant properties of lycopene (Lyc), selenium nanoparticles (Se-NPs), and lycopene selenium nanoparticles (Lyc-Se-NPs). Methods: FTIR, polydispersity index, and zeta potential evaluations provided a complete characterization of the synthesized Lyc-Se-NPs. The broth dilution method and a crystal violet microtiter plate assay were employed to assess the antibacterial and antibiofilm activity, respectively. The rat wound infection model was performed to study the anti-inflammatory effect. Findings: The Lyc-Se-NPs had a zeta potential range of −16.93 to −31.04 mV and a mean particle size of 126.6 ± 3.12 nm. All peaks’ percentage transmittance decreased, according to the FTIR analysis of the Lyc-Se-NPs, with the exception of one peak at 2924.22 cm−1, which is suggestive of C-H stretching. The mean scavenging concentrations for Lyc-Se-NPs in the DPPH and ABTS radical scavenging experiments were 3.85 ± 0.65 and 4.26 ± 0.7 µg/mL, respectively. For S. aureus, the Lyc-Se-NPs’ MIC values varied from 64 to 1024 µg/mL. CLSM verified that S. aureus treated with sub-MICs of Lyc-Se-NPs showed a significant reduction in biofilm formation. Furthermore, the group treated with 50 mg of Lyc-Se-NPs showed the quickest rate of wound healing. They demonstrated a notable elevation of the HO−1 content in skin tissues, together with the greatest downregulation of TNF-α, IL-1β, and COX-2. Conclusions: The distinguishing features of Lyc-Se-NPs reveal that this unique compound is a promising antibacterial, antioxidant, and anti-inflammatory agent.

Pharmaceuticals doi: 10.3390/ph17121599

Authors: Qinyi Li Xiaohong Zhao Huan Yang Xiaolong Zhu Xinbing Sui Jiao Feng

Endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) play critical roles in tumorigenesis, cancer progression, and drug resistance. Persistent activation of the ER stress system enhances the survival capacities of malignant tumor cells, including increased proliferation, invasion, and resistance to treatment. Dysregulation of ER function and the resultant stress is a common cellular response to cancer therapies and may lead to cancer cell death. Currently, growing evidence suggests that Traditional Chinese medicine (TCM), either as a monotherapy or in combination with other treatments, offers significant advantages in preventing cancer, inhibiting tumor growth, reducing surgical complications, improving drug sensitivity, and mitigating drug-induced damage. Some of these natural products have even entered clinical trials as primary or complementary anticancer agents. In this review, we summarize the anticancer effects of TCM monomers/natural products on the gastrointestinal (GI) tumors and explore their mechanisms through ER stress modulation. We believe that ongoing laboratory research and the clinical development of TCM-based cancer therapies hold considerable potential for advancing future cancer treatments.

Pharmaceuticals doi: 10.3390/ph17121598

Authors: Umme Hani Sharanya Paramshetti Mohit Angolkar Wajan Khalid Alqathanin Reema Saeed Alghaseb Saja Mohammed Al Asmari Alhanouf A. Alsaab Farhat Fatima Riyaz Ali M. Osmani Ravi Gundawar

Background: Psoriatic arthritis (PsA), a chronic inflammatory disease, mainly affects the joints, with approximately 30% of psoriasis patients eventually developing PsA. Characterized by both innate and adaptive immune responses, PsA poses significant challenges for effective treatment. Recent advances in drug delivery systems have sparked interest in developing novel formulations to improve therapeutic outcomes. The current research focuses on the development and evaluation of a nanosponge-loaded, cyclo-oxygenase-2 (COX-2) inhibitor-based topical gel for the treatment of PsA. Methods: Nanosponges (NSs) were prepared by using beta-cyclodextrin as a polymer and dimethyl carbonate (DMC) as a crosslinker by melting, and gels were prepared by employing carbopol and badam gum as polymers. Results: Solubility studies confirmed that the prepared nanosponges were highly soluble. FT-IR studies confirmed the formation of hydrogen bonds between lumiracoxib and beta-cyclodextrin. SEM confirmed that the prepared formulations were roughly spherical and porous in nature. The average particle size was 190.5 ± 0.02 nm, with a zeta potential of −18.9 mv. XRD studies showed that the crystallinity of lumiracoxib decreased after encapsulation, which helped to increase its solubility. The optimized nanosponges (NS2) were incorporated in an optimized gel (FG10) to formulate a nanosponge-loaded topical gel. The optimized gel formulation exhibited a homogeneous consistency, with a pH of 6.8 and a viscosity of 1.15 PaS, indicating its suitability for topical application and stability. The in vitro diffusion studies for the topical gel showed drug release of 82.32% in 24 h. The optimized formulation demonstrated significant antipsoriatic activity, as confirmed through cytotoxicity studies conducted on HaCaT cells. Conclusions: On the basis of the findings, it can be concluded that the prepared nanosponge-loaded topical gel formulation presents a promising solution for the effective management of PsA, offering enhanced drug solubility, sustained release, and improved therapeutic potential.

Pharmaceuticals doi: 10.3390/ph17121597

Authors: Alessandro Medoro Giovanni Scapagnini Simone Brogi Tassadaq Hussain Jafar Truong Tan Trung Luciano Saso Sergio Davinelli

Background/Objectives: This study investigated the potential of green algae-derived carotenoids as natural inhibitors of the proprotein convertase subtilisin/kexin type 9 (PCSK9), a key regulator of cholesterol metabolism. PCSK9 promotes the degradation of low-density lipoprotein receptors (LDLR), thereby increasing blood cholesterol levels and elevating the risk of cardiovascular diseases. Methods/Results: We screened the pharmacophore fit score of 27 carotenoids with PCSK9 and identified 14 that were analyzed for binding affinity and molecular interactions. Astaxanthin, siphonaxanthin, and prasinoxanthin were identified as the top candidates, demonstrating strong binding affinity (−10.5, −10.3, and −9.4 Kcal/mol, respectively) and stable interactions with several known key residues within the active site of PCSK9, including Pro-331, Arg-357, Cys-358, Val-359, Asp-360, Ile-416, Leu-436, Thr-437, Pro-438, Leu-440, Arg-458, Val-460, Trp-461, Arg-476, Cys-477, Ala-478, Ala-649, Val-650, and Asp-651. Density functional theory analysis confirmed the stability of astaxanthin and its favorable electronic properties, suggesting its potential as an effective inhibitor. Molecular dynamics simulations of the PCSK9–astaxanthin complex revealed sustained structural stability and key interactions critical for maintaining the functional integrity of the protein. Conclusions: These findings provide evidence that specific carotenoids, particularly astaxanthin, may offer a cost-effective alternative to existing PCSK9 inhibitors, providing a potential approach for managing cholesterol levels and reducing cardiovascular risk. Pre-clinical and clinical validations are required to confirm the therapeutic potential of these compounds.

Pharmaceuticals doi: 10.3390/ph17121596

Authors: Anna Stasiłowicz-Krzemień Julia Wójcik Anna Gościniak Marcin Szymański Piotr Szulc Krzysztof Górecki Judyta Cielecka-Piontek

Background: Curcuminoids, the bioactive compounds found in turmeric, exhibit potent antioxidant, anti-inflammatory, and neuroprotective properties. This study aims to enhance the extraction of curcuminoids from turmeric using environmentally friendly solvents supercritical CO2 (scCO2) combined with natural deep eutectic solvents (NADESs) in one process, and to evaluate the resulting biological activity. Methods: A Box–Behnken statistical design was applied to optimize scCO2 extraction conditions—pressure, CO2 volume, and temperature—to maximize curcuminoid yield. Next, the menthol and lactic acid NADESs were selected, and these two solvents were combined into a single turmeric extraction process. The biological activity of the resulting extract was evaluated using antioxidant assays (ferric reducing antioxidant power and 2,2-diphenyl-1-picrylhydrazyl) and enzyme inhibition assays (acetylcholinesterase, butyrylcholinesterase, and tyrosinase). Toxicity assessments were conducted on the aquatic invertebrates Daphnia pulex, Artemia sp., and Chironomus aprilinus. Results: The most effective extraction was achieved using a menthol–lactic acid NADES as a cosolvent, integrated at a 1:20 ratio of plant material to NADESs while in combination with scCO2. The optimized scCO2–NADES extraction resulted in a high curcuminoid yield (33.35 mg/g), outperforming scCO2 extraction (234.3 μg/g), NADESs ultrasound-assisted extraction (30.50 mg/g), and alcohol-based solvents (22.95–26.42 mg/g). In biological assays, the extract demonstrated significant antioxidant activity and effective inhibition of enzymes (acetylcholinesterase, butyrylcholinesterase, and tyrosinase). Toxicity studies showed a concentration-dependent response, with EC50 for Chironomus aprilinus at the level of 0.098 μL/mL and Daphnia pulex exhibiting high sensitivity to the extract. Conclusions: This study highlights the potential of combining NADESs and scCO2 extraction in one process, demonstrating the effectiveness of scCO2–NADES extraction in maximizing curcuminoid yield and enhancing bioactivity.

Pharmaceuticals doi: 10.3390/ph17121595

Authors: Edgar Daniel Moyers-Montoya María Jazmín Castañeda-Muñoz Daniel Márquez-Olivas René Miranda-Ruvalcaba Carlos Alberto Martínez-Pérez Perla E. García-Casillas Wilber Montejo-López María Inés Nicolás-Vázquez René Gerardo Escobedo-González

Background/Objectives: Breast cancer is a disease with a high mortality rate worldwide; consequently, urgent achievements are required to design new greener drugs, leaving natural products and their derivatives as good options. A constant antineoplastic effect has been observed when the phytoproduct contains an indole fragment. Methods: Therefore, the objective of this work was to carry out a thoughtful computational study to perform an appropriate evaluation of four novel molecules of the class of the 3-indolylquinones as phytodrug candidates for antineoplastic activity: thymoquinone (TQ), 2,6-dimethoxy-1,4-benzoquinone (DMQ), 2,3-dimethoxy-5-methyl-1,4-benzoquinone (DMMQ), and 2,5-dihydroxy-1,4-benzoquinone (DHQ). It is important to highlight that the obtained computational results of the _target compounds were compared-correlated with the theoretical and experimental literature data previously reported of several indolylquinones: indolylperezone, indolylisoperezone, indolylmenadione, and indolylplumbagin (IE-IH, respectively). Results: The results revealed that the studied structures possibly presented antineoplastic activity, in addition to the fact that the reactivity parameters showed that two of the evaluated compounds have the option to present IC50 values lower than or similar to 25 mg/mL, activity like that of indolylisoperezone; moreover, they show molecular coupling to PARP-1. Finally, the prediction of the calculated physicochemical parameters coincides with the Lipinski and Veber rules, indicating that the adsorption, metabolism, and toxicity parameters are acceptable for the studied compounds, obtaining high drug score values. Conclusions: Finally, a comparison between the proposed molecules and others previously synthesized was appropriately performed, establishing that the synthesis of the studied compounds and the determination of their pharmacological properties in an experimental manner are of interest.

Pharmaceuticals doi: 10.3390/ph17121594

Authors: Daniel Ricardo Delgado Jennifer Katiusca Castro-Camacho Claudia Patricia Ortiz Diego Ivan Caviedes-Rubio Fleming Martinez

Background: Solubility is one of the most important parameters in the research and development processes of the pharmaceutical industry. In this context, cosolubility is one of the most used strategies to improve the solubility of poorly soluble drugs, besides allowing to identify some factors involved in the dissolution process. The aim of this research is to evaluate the solubility of sulfamethazine in acetotinitrile + 1-propanol cosolvent mixtures at 9 temperatures (278.15, 283.15, 288.15, 293.15, 298.15, 303.15, 308.15, 313.15, and 318.15 K); a drug used in human and veterinary therapy and two solvents of great chemical–pharmaceutical interest. Methods: The determination was carried out by the shaking flask method and the drug was quantified by UV/Vis spectrophotometry. Results: The solubility of sulfamethazine increases from pure 1-propanol (solvent in which it reaches its lowest solubility at 278.15 K) to pure acetonitrile (solvent in which it reaches its maximum solubility at 318.15 K), behaving in a logarithmic-linear fashion. Conclusions: The increase in solubility is related to the acid/base character of the cosolvent mixtures and not to the solubility parameter of the mixtures. The dissolution process is endothermic and favored by the solution entropy, and also shows a strong entropic compensation.

Pharmaceuticals doi: 10.3390/ph17121593

Authors: Octavia-Laura Oancea Șerban Andrei Gâz Gabriel Marc Ioana-Andreea Lungu Aura Rusu

Background/Objectives: Fluoroquinolones (FQs) are topoisomerase II inhibitors with antibacterial activity, repositioned recently as anti-cancer agents. Glutamic acid (GLA) is an amino acid that affects human metabolism. Since an anti-cancer mechanism of FQs is human topoisomerase II inhibition, it is expected that FQ-GLA hybrids can act similarly. Methods: We designed 27 hypothetical hybrids of 6 FQs and GLA through amide bonds at the 3- and 7-position groups of FQs or via ethylenediamine/ethanolamine linkers at the carboxyl group of the FQ. Hydroxamic acid derivatives were also theoretically formulated. Computational methods were used to predict their physicochemical, pharmacokinetic, or toxicological properties and their anti-cancer activity. For comparison, etoposide was used as an anti-cancer agent inhibiting topoisomerase II. Molecular docking assessed whether the hybrids could interact with the human topoisomerase II beta in the same binding site and interaction sites as etoposide. Results: All the hybrids acted as potential topoisomerase II inhibitors, demonstrating possible anti-cancer activity on several cancer cell lines. Among all the proposed hybrids, MF-7-GLA would be the ideal candidate as a lead compound. The hybrid OF-3-EDA-GLA and the hydroxamic acid derivatives also stood out. Conclusions: Both FQs and GLA have advantageous structures for obtaining hybrids with favourable properties. Improvements in the hybrids’ structure could lead to promising results.

Pharmaceuticals doi: 10.3390/ph17121592

Authors: Katarina Dathe Carolin Benndorf Simone Bergner Christof Schaefer

Background: Paracetamol and non-steroidal anti-inflammatory drugs (NSAIDs) are frequently used during pregnancy. Due to their fetotoxicity, NSAIDs are contraindicated during the third trimester. There is ongoing controversy about the extent to which NSAIDs may cause cardiovascular and renal impairment in the fetus earlier in the second trimester. Paracetamol, used as an effective treatment for closure of patent ductus arteriosus (PDA) after birth, is suspected to cause similar but unwanted effects during the third trimester of pregnancy. Methods: Three major pharmacovigilance databases (VigilanceCentral, EudraVigilance, and VigiBase) were searched for Individual Case Safety Reports (ICSRs; n = 1288) on fetotoxic effects that have been shown to result from NSAID exposure in late pregnancy. Results: In 219/1288 cases, an NSAID and/or paracetamol was taken after the first trimester, and the ICSR was not related to other reported risk factors. Out of these 219 ICSRs, 48 were exposed to NSAIDs in the second but not the third trimester or to paracetamol in the third trimester. Causality assessment was “probable or likely” in four NSAID reports and none of the paracetamol reports. Conclusions: The scarcity of adverse drug reactions (ADRs) in our study and in the literature, despite decades of pharmaceutical marketing and worldwide use of paracetamol as an analgesic of choice in the third trimester and the absence of formal contraindications against NSAIDs in the second trimester, speaks against a substantial cardiovascular and nephrotoxic risk of temporary use of NSAIDs in the second trimester or paracetamol in the third trimester. NSAIDs continue to be contraindicated in the third trimester.

Pharmaceuticals doi: 10.3390/ph17121591

Authors: Fatheia N. Hamza Khalid Said Mohammad

Bone metastases are a prevalent complication in advanced cancers, particularly in breast, prostate, and lung cancers, and are associated with severe skeletal-related events (SREs), including fractures, spinal cord compression, and debilitating pain. Conventional bone-_targeted treatments like bisphosphonates and RANKL inhibitors (denosumab) reduce osteoclast-mediated bone resorption but do not directly impact tumor progression within the bone. This review focuses on examining the growing potential of immunotherapy in _targeting the unique challenges posed by bone metastases. Even though immune checkpoint inhibitors (ICIs) have significantly changed cancer treatment, their impact on bone metastases appears limited because of the bone microenvironment’s immunosuppressive traits, which include high levels of transforming growth factor-beta (TGFβ) and the immune-suppressing cells, such as regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs). This review underscores the investigation of combined therapeutic approaches that might ease these difficulties, such as the synergy of immune checkpoint inhibitors with agents aimed at bones (denosumab, bisphosphonates), chemotherapy, and radiotherapy, as well as the combination of immune checkpoint inhibitors with different immunotherapeutic methods, including CAR T-cell therapy. This review provides a comprehensive analysis of preclinical studies and clinical trials that show the synergistic potential of these combination approaches, which aim to both enhance immune responses and mitigate bone destruction. By offering an in-depth exploration of how these strategies can be tailored to the bone microenvironment, this review underscores the need for personalized treatment approaches. The findings emphasize the urgent need for further research into overcoming immune evasion in bone metastases, with the goal of improving patient survival and quality of life.

Pharmaceuticals doi: 10.3390/ph17121590

Authors: Elias Leiva-Salcedo Denise Riquelme Juan Pablo Huidobro-Toro Claudio Coddou

Background/Objectives: P2X receptor channels are widely expressed in the CNS, where they have multiple functions in health and disease. The rat P2X2a (rP2X2a) receptor channel is modulated by copper, an essential trace element that plays important roles in synaptic modulation and neurodegenerative disorders. Although essential extracellular amino acids that coordinate copper have been identified, the exact mechanism of copper-induced modulation has not been yet elucidated. Methods: We used HEK293T cells expressing rP2X2a channel(s) and performed outside-out single-channel and whole-cell recordings to explore copper’s effects on rP2X2 currents and determine whether this metal can increase the cooperative gating of rP2X2a channel. Results: In whole-cell recordings and in patches containing 2 or 3 rP2X2a channels, copper enhanced the ATP-induced currents, significantly reducing the ATP EC50 and increasing the Hill coefficient. Moreover, copper increased the apparent Po in patches containing two or three channels. By contrast, in patches containing only one rP2X2a channel, we did not observe any significant changes in ATP EC50, the Hill coefficient, or Po. Conclusions: Copper modulates the gating of rP2X2a channels, enhancing interchannel cooperativity without altering single-channel conductance or Po. This novel regulatory mechanism could be relevant for understanding the role of P2X2 channels in physiological and pathological processes.

Pharmaceuticals doi: 10.3390/ph17121589

Authors: Jieun Park Jaehong Kim

Prostate cancer (PCa) is the most prevalent malignancy and the second leading cause of cancer-related death in men. Although current therapies can effectively manage the primary tumor, most patients with late-stage disease manifest with metastasis in different organs. From surgery to treatment intensification (TI), several combinations of therapies are administered to improve the prognosis of patients with metastatic PCa. Due to the high frequency of the mutation during the metastatic phase, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) genetic engineering tool can accelerate the effects of TI by enhancing _targeted gene therapy or immunotherapy. This review describes the genetic background of metastatic PCa and how CRISPR/Cas9 technology can contribute to the field of PCa treatment development. It also discusses the current limitations of conventional PCa therapy and the potential of CRISPR-based PCa therapy.

Pharmaceuticals doi: 10.3390/ph17121588

Authors: Jan Bures Martin Novak Vera Radochova Darina Kohoutova Lukas Prchal Jan Martinek Jan Mares Jaroslav Cerny Stepan Suchanek Jaroslav Pejchal Barbora Voxova Petr Urbanek Miroslav Zavoral Ondrej Soukup

Background/Objectives: Tacrine is a centrally active non-competitive reversible acetylcholinesterase inhibitor. It also exerts antagonising activity against N-methyl-D-aspartate receptors. Tacrine was approved for the treatment of Alzheimer’s disease in 1993, but was withdrawn from clinical use in 2013 because of its hepatotoxicity and gastrointestinal side effects. Nevertheless, tacrine is currently facing a renewed wave of interest primarily due to several new tacrine-incorporated hybrids and derivates. There were two specific aims for this study: firstly, to explain the mechanisms of the adverse action of tacrine, as a distinctive example of a highly effective acetylcholinesterase inhibitor; and secondly to check whether luminal impedance planimetry is feasible for preclinical testing of possible side effects of compounds potentially toxic to the gastrointestinal tract. Methods: Six experimental pigs were used as the animal model in this study. Five major parameters were evaluated: luminal pressure (mmHg), estimated diameter (mm), cross-sectional area (mm2), distensibility (mm2/mmHg), and zone compliance (mm3/mmHg). All measurements were performed before and 360 min after intragastric administration of 200 mg tacrine (at the porcine tacrine Tmax). Results: This study consistently demonstrated an increase in luminal pressure (a directly measured indicator) for the particular balloon filling volumes used, and inversely a reciprocal decrease in the other parameters after tacrine administration. Conclusions: Endoscopic luminal impedance planimetry is a feasible method to evaluate functional response of the lower oesophageal sphincter to tacrine in experimental pigs. Tacrine did not compromise the function of the lower oesophageal sphincter either toward oesophageal spasms or, in contrast, decreased competence of the lower oesophageal sphincter.

Pharmaceuticals doi: 10.3390/ph17121587

Authors: Ursula Wolf Luise Drewas Hassan Ghadir Christian Bauer Lars Becherer Karl-Stefan Delank Rüdiger Neef

Background/Objectives: Falls and fractures are emerging as a near-pandemic and major global health concern, placing an enormous burden on ageing patients and public health economies. Despite the high risk of polypharmacy in the elderly patients, falls are usually attributed to age-related changes. For the “Individual Pharmacotherapy Management (IPM)” established at the University Hospital Halle, the IPM medication adjustments and their association with in-hospital fall prevention were analysed. Methods: On the basis of the most updated digital overall patient view via his inpatient electronic health record (EHR), IPM adapts each drug’s Summary of Product Characteristics to the patient’s condition. A retrospective pre-post intervention study in geriatric traumatology on ≥70 years old patients compared 200 patients before IPM implementation (CG) with 204 patients from the IPM intervention period (IG) for the entire medication list, organ, cardiovascular and vital functions and fall risk parameters. Results: Statistically similar baseline data allowed a comparison of the average 80-year-old patient with a mean of 11.1 ± 4.9 (CG) versus 10.4 ± 3.6 (IG) medications. The IPM adjusted for drug-drug interactions, drug-disease interactions, overdoses, anticholinergic burden, adverse drug reactions, esp. from opioids inducing increased intrasynaptic serotonin, psychotropic drugs, benzodiazepines, contraindications and missing prescriptions. IPM was associated with a significant reduction in in-hospital falls from 18 (9%) in CG to 3 (1.5%) in IG, a number needed to treat of 14, relative risk reduction 83%, OR 0.17 [95% CI 0.04; 0.76], p = 0.021 in multivariable regression analysis. Factors associated with falls were antipsychotics, digitoxin, corticosteroids, Würzburg pain drip (combination of tramadol, metamizole, metoclopramide), head injury, cognitive impairment and aspects of the Huhn Fall Risk Scale including urinary catheter. Conclusion: The results indicate medication risks constitute a major iatrogenic cause of falls in this population and support the use of EHR-based IPM in standard care for the prevention of falls in the elderly and for patient and drug safety. In terms of global efforts, IPM contributes to the running WHO and United Nations Decade of Healthy Ageing (2021–2030).

Pharmaceuticals doi: 10.3390/ph17121585

Authors: Kenzi Riboulet-Zemouli Josep Allué Creus

This study explores the trajectory of interest in and use of Extractum Cannabis (hemp extract, i.e., extract of Cannabis sativa L.) for the symptomatic treatment of minor gastrointestinal distress and dyspepsia in nineteenth- and early twentieth-century Barcelona (Catalonia, Spain) prior to 1939, through a review of primary sources. The objective of this paper is to present a historical pharmaceutical and applied review of the medical use of the hemp genus (Cannabis L.) prior to its prohibition, thereby contributing to its recognition as a medicinal product. The information provided demonstrates evidence of the medicinal use of cannabis within the historical context studied. The interactions between this legacy medical use and the contemporary body of pharmacological and toxicological knowledge (on hemp, its constituents, and the endocannabinoid system in gastrointestinal and stomach disorders) are discussed, providing new possible clinical perspectives. Within its limitations—including the scope, limited accessibility to, and varying quality of archives—this research contributes to a more granular understanding of the historical embeddedness of psychoactive hemp medicines in northeastern Spain, suggesting that medical and pharmaceutical traditions could play a role in informing contemporary approaches to “medical marijuana”.

Pharmaceuticals doi: 10.3390/ph17121586

Authors: Elitsa L. Pavlova Elena P. Nenova Lyubomira D. Yocheva Iliana A. Ivanova Peter A. Georgiev

Objectives: The antimicrobial, oxidative activities, and ecotoxicity of synthesized silver-loaded zeolites (X and ZSM-5(MFI), Si-to-Al ratios 12 and 25) were studied, linking antimicrobial properties to material structure and released active silver species. Methods: The materials were characterized by SEM, EDX, TEM, and XRPD. All materials, with a silver content of 1–3%wt for the Ss and about 35%wt for the X-zeolites, were tested against Escherichia coli and Staphylococcus aureus. Redox activity was studied in physiological (pH 7.4/37 °C) and optimal (pH 8.5/37 °C) conditions in chemiluminescent model systems. In the ecotoxicity tests, we used Daphnia magna. Results: A proportional correlation was observed between the bactericidal effect of and the silver content in the zeolites. AgX with a Si/Al ratio of ~1.23 and 35% silver showed a higher antimicrobial efficiency, particularly against Gram-negative E. coli versus Gram-positive S. aureus. The concentration thresholds were as follows: AgXas had a bactericidal effect at 0.003 g/L−1, with an MIC at 0.0015 m/L−1 for E. coli; SA25-Ag, AgXcl, AgXrc had a bactericidal effect at 2.5 g/L−1. The bacteria were more resilient than Daphnia magna, which showed a 90–100% lethality at Ag–zeolite concentrations of 0.00625 to 0.0125 g/L−1. AgXas and AgXrc demonstrated strong reactive oxygen species generation at both the physiological and optimal pH, explaining their bactericidal effects. In general, the tested materials showed an inhibition of the generated reactive oxygen species depending on the model system and conditions. Conclusions: The silver species leached from the new materials explain their higher oxidation and bactericidal activity. While suitable for stringently controlled biological applications, their release into the environment, in concentrations higher than 0.01g/L−1, should be avoided.

Pharmaceuticals doi: 10.3390/ph17121584

Authors: Rupert M. Bauersachs Edelgard Lindhoff-Last Robert Klamroth Andreas Koster Marc Schindewolf Harry Magnani

(1) Background: Danaparoid sodium is a heparinoid antithrombotic that has been used for over 40 years for prophylaxis of DVT in non-HIT patients and for the treatment of heparin-induced thrombocytopenia (HIT) with and without thrombosis. This update summarises current information on its pharmacology and reviews danaparoid dose management in a broad spectrum of clinical situations, including off-label indications. (2) Methods: Evidence from published clinical studies, case reports, compassionate use of danaparoid, and spontaneously reported serious adverse events is summarised and analysed by an interdisciplinary expert group to develop a consensus on dosing regimens of danaparoid for complex clinical situations, including vulnerable patient populations. (3) Results: Dosing regimens are proposed, together with monitoring recommendations and _target anti-factor Xa ranges. (4) Conclusion: In a comprehensive summary detailed interdisciplinary dosing recommendations are described to provide a basis for safe and effective use of danaparoid.

Pharmaceuticals doi: 10.3390/ph17121583

Authors: Lei Zheng Biao Zhao Zhenxi Zhang Yutong Liu Yingying Zhang Jing Cai Tong Qiao

Background/Objectives: Chronic Limb-Threatening Ischemia (CLTI) is a chronic limb ischemic disease caused by vascular lesions, characterized by pain, ulcers, and gangrene, which can be life-threatening in severe cases. The objective of this study is to explore whether Berbamine (BBM) can protect against and repair ischemic muscle tissue in the lower limbs; Methods: Using a mouse hindlimb ischemia (HLI) model, 36 C57BL6 mice were divided into sham, HLI, and HLI+BBM treatment groups. Results: Our findings indicate that BBM can restore motor function and muscle tissue pathology in mice, potentially by inhibiting the nuclear translocation of nuclear factor kappa-B (NF-κB), thereby alleviating tissue inflammation caused by chronic ischemia, reducing muscle cell apoptosis, inhibiting M1 macrophage polarization, and promoting angiogenesis. Conclusions: Our research suggests that BBM has the potential to protect against ischemic damage in lower limb muscle tissue, providing a new approach to the treatment of CLTI.

Pharmaceuticals doi: 10.3390/ph17121582

Authors: Anton A. Kornev Stanislav V. Shmakov Alexander I. Ponyaev Alexander V. Stepakov Vitali M. Boitsov

Background: A series of spiro-fused heterocyclic compounds containing cyclopropa[a]pyrrolizidine-2,3′-oxindole and 3-spiro[3-azabicyclo[3.1.0]-hexane]oxindole frameworks were synthesized and studied for their in vitro antiproliferative activity against human erythroleukemia (K562), cervical carcinoma (HeLa), acute T cell leukemia (Jurkat), melanoma (Sk-mel-2) and breast cancer (MCF-7) as well as mouse colon carcinoma (CT26) cell lines. Methods: Cell proliferation was evaluated in vitro by MTS assay. Confocal microscopy was used to study actin cytoskeleton structure and cell motility. Cell cycle analysis was evaluated by flow cytometry. Results: It was found that compounds 4, 8, 18 and 24 showed antiproliferative activity against the Jurkat, K-562, HeLa and Sk-mel-2 cell lines with IC50 ranging from 2 to 10 μM (72 h). Evaluation of the impact on cell cycle progression showed that the tested compounds achieved significant cell-cycle perturbation with a higher accumulation of cells in the SubG1 and G0/G1 phases of the cell cycle, in comparison to the negative control. I Incubation with tested compounds led to the disappearance of stress fibers (granular actin was distributed diffusely in the cytoplasm in up to 38% of treated HeLa cells) and changes in the number of filopodia-like deformations (reduced from 93% in control cells to 64% after treatment). The impact on the Sk-mel-2 cell actin cytoskeleton structure was even greater: granular actin was distributed diffusely in the cytoplasm in up to 90% of treated cells while the number of filopodia-like deformations was reduced by up to 23%. A scratch test performed on the human melanoma cell line showed that these cells did not fill the scratched strip and lose their ability to move under treatment. Conclusions: The obtained results support the antitumor effect of the tested spiro-compounds and encourage the extension of this study in order to improve their anticancer activity as well as reduce their toxicological risks.

Pharmaceuticals doi: 10.3390/ph17121581

Authors: Maram H. Abduljabbar

This paper investigates the therapeutic use of PCSK9 inhibitors, particularly Evolocumab, as monoclonal antibodies for the treatment of atherosclerosis based on recent literature reviews. PCSK9 is an outstanding example of a breakthrough in medical science, with advancements in understanding its biological function driving substantial progress in atherosclerosis treatment. Atherosclerotic cardiovascular disease (ASCVD) is a leading global cause of mortality, imposing substantial financial burdens on healthcare systems. Elevated low-density lipoprotein cholesterol (LDL-C), a modifiable risk factor, plays a pivotal role in the development of ASCVD. Emerging treatments such as PCSK9 inhibitors are now being introduced to combat this issue, with the goal of reducing ASCVD risk by directly _targeting LDL-C levels. This discovery highlighted the potential of monoclonal antibodies to inhibit PCSK9, thereby enhancing LDL-C receptor activity. This breakthrough led to the development of Alirocumab and Evolocumab inhibitors, which typically reduce LDL-C levels by approximately 50%. This research underscores the importance of PCSK9 inhibitors in treating ASCVD, drawing on evidence from various randomized controlled trials such as FOURIER, ODYSSEY OUTCOMES, and VESALIUS-CV. These trials have also shown that PCSK9 inhibitors are effective and safe for the treatment of several cardiovascular disorders. PCSK9 inhibitors are therefore useful in patients who do not reach their _target LDL-C levels when on the highest doses of statins or patients with very high cardiovascular risk who cannot tolerate statins at all.

Pharmaceuticals doi: 10.3390/ph17121580

Authors: Dhafer Alwayli Xiaoli Jiang Jiaxu Liang Syed Rafiq Hussain Shah Atta Ullah Mohammed F. Z. Abusidu Wen Shu

Background: Sublingual immunotherapy (SLIT) has shown promise in mitigating allergic asthma symptoms; nevertheless, its high dose and prolonged duration of treatment raise safety concerns. This study explored the potential of Lactobacillus paracasei (L. paracasei) to enhance the effectiveness of SLIT in a mouse model of allergic asthma. Methods: Allergic asthma was induced in Balb/c mice following sensitization and challenge with a house dust mite (HDM) allergen. Subsequently, the mice were subjected to SLIT (66 and 132 µg) either alone or in combination with L. paracasei supplementation. Asthma-associated parameters, including rubbing frequency, IgE level, cytokine profiles, and histological changes, were evaluated to assess treatment efficacy. Results: mice that received SLIT 132 µg combined with the probiotic (combined 132) demonstrated a significant reduction in allergic symptoms (rubbing). This treatment strategy led to a marked IgE and eosinophil level decrease in serum; an increase in anti-inflammatory cytokines like IFN-γ and IL-10; and a reduction in pro-inflammatory cytokines IL-17 and TNF-α. The combination therapy also mitigated lung inflammation and supported the restoration of the structural integrity of the colon, promoting the recovery of goblet cells and mucus secretion. Probiotic treatment alone also effectively reduced IgE levels, increased IFN-γ, and decreased levels of IL-17 and TNF-α. Conclusions: The adjuvant effect of L. paracasei in enhancing SLIT represents a promising approach for improving asthma treatment efficacy.

Pharmaceuticals doi: 10.3390/ph17121579

Authors: Cristina Hernández Hugo Ramos Anne Létondor Rafael Simó

Background/Objectives: Early stages of diabetic retinopathy are currently considered an unmet medical need due to the lack of effective treatments beyond proper monitoring and control of glycemia and blood pressure. Sitagliptin eye drops have emerged as a new therapeutic approach against early stages of the disease, as they can prevent its main hallmarks, including both neurodegeneration and microvascular impairment. Interestingly, all of these effects occur without any glycemic systemic improvement. In the present study, we aimed to investigate the pharmacokinetics and distribution of the drug within the eye and plasma. Methods: A total of 48 male New Zealand rabbits were treated with topical administration (eye drops) of sitagliptin at two concentrations: 5 mg/mL and 10 mg/mL. Blood, iris/ciliary body, retina/choroid, aqueous humor, and vitreous humor samples were collected at specific intervals post-administration (10 and 30 min and 1, 3, 6, 15, and 24 h), processed, and analyzed using an LC-MS/MS method. The pharmacokinetics of sitagliptin were then calculated, and statistical comparisons were performed. Results: Our findings indicate that sitagliptin reaches the retina prior to the aqueous and vitreous humors, suggesting that its absorption follows the transscleral route. Additionally, systemic absorption was minimal and below pharmacologically active concentrations. Conclusions: These results support the use of an eye drop formulation for the treatment of diabetic retinopathy and other retinal diseases.

Pharmaceuticals doi: 10.3390/ph17121578

Authors: Redouane Tarik Aziz Drioiche Jalila El Amri Mohamed Ed-Dahmouny Abdelaaty Abdelaziz Shahat Nadia Hadi Mouradi Aicha Handaq Nadia Fadoua El Makhoukhi Abdelhakim El Ouali Lalami Noureddine Elmoualij Eto Bruno Hajji Lhoussain Touriya Zair

Background: Teucrium capitatum L., a member of the Lamiaceae family, is widely used in traditional medicine for its therapeutic properties. This study aims to analyze the chemical composition of its essential oil and extracts, evaluate their antimicrobial and antioxidant activities, and investigate the interactions of their bioactive compounds with biological _targets using in silico methods to better understand their mechanisms of action. Methods: Essential oil was extracted via hydrodistillation from leaves collected in Morocco, while phenolic compounds were obtained through Soxhlet and decoction extraction methods. Gas chromatography-mass spectrometry (GC-MS) was used for chemical profiling. Antimicrobial and antioxidant activities were assessed using standard methods, including DPPH, FRAP, and TAC assays. Molecular docking was conducted to explore interactions between major constituents and biological _targets. Results: GC-MS analysis revealed significant bioactive components in the essential oil, such as β-pinene (24.5%), α-cadinol (17.02%), and shyobunol (12.13%). Extracts (hydro-ethanolic, hydro-methanolic, and aqueous via decoction) were rich in poliumoside (27.74%) and cirsimaritin (28.22%). The essential oil and extracts showed significant antimicrobial activity, particularly against Staphylococcus aureus, Escherichia coli, Candida albicans, and Aspergillus niger. Antioxidant assays confirmed strong activity. Molecular docking results supported strong interactions of major compounds with key biological _targets. Conclusions: The high presence of phenolic and flavonoid compounds in Teucrium capitatum extracts contributes to their strong antimicrobial and antioxidant properties, supporting their potential for development as natural therapeutic agents.

Pharmaceuticals doi: 10.3390/ph17121577

Authors: Seulgi Shin Hyejung Jo Tomoyo Agura Seoyoun Jeong Hyovin Ahn Soyoung Pang June Lee Jeong-Ho Park Yejin Kim Jae Seung Kang

Background/Objectives: Vitamin C is a well-known antioxidant with antiviral, anticancer, and anti-inflammatory properties. However, its therapeutic applications are limited by rapid oxidation due to heat and light sensitivity. Aptamin C, which employs aptamers to bind vitamin C, has demonstrated enhanced stability and efficacy. This study investigates the potential of Aptamin C to inhibit the progression of pulmonary fibrosis, a prominent inflammatory lung disease with no effective treatment. Methods: Mice bearing bleomycin-induced pulmonary fibrosis were administered vitamin C or Aptamin C, and their weight changes and survival rates were monitored. Inflammatory cell infiltration was assessed in the bronchoalveolar lavage fluid (BALF), and the degree of alveolar fibrosis was measured by H&E and Masson’s trichrome staining. To elucidate the mechanism of action of Aptamin C, Western blot analysis was performed in HaCaT and lung tissues from bleomycin-induced pulmonary fibrosis mice. Results: The Aptamin C-treated group showed a notably higher survival rate at 50%, whereas all subjects in the vitamin C-treated group died. Histological examination of lung tissue showed that inflammation was significantly suppressed in the Aptamin C-supplemented group compared to the vitamin C-supplemented group, with a 10% greater reduction in cell infiltrations, along with noticeably less tissue damage. Additionally, it was observed that Aptamin C increased SVCT-1 expression in the HaCaT cells and the lung tissues. Conclusions: Taken together, Aptamin C not only increases the stability of vitamin C but also induces an increase in SVCT-1 expression, facilitating greater vitamin C absorption into cells and tissues, thereby inhibiting the progression of symptoms and associated inflammatory responses in pulmonary fibrosis.

Pharmaceuticals doi: 10.3390/ph17121576

Authors: Mike Wagenbrenner Tizian Heinz Philip M. Anderson Ioannis Stratos Joerg Arnholdt Susanne Mayer-Wagner Konstantin Horas Denitsa Docheva Boris M. Holzapfel Maximilian Rudert Manuel Weißenberger

Background: The aim of our study was to examine the combined effects of tranexamic acid (TXA) and vancomycin powder (VP) on chondrocytes in vitro. Despite the use of TXA and VP being linked to a reduced risk of extensive postoperative blood loss and periprosthetic joint infections (PJIs) in TKA, the possible cytotoxic side effects on periarticular cell types remain unclear. Methods: Human chondrocytes were harvested from hyaline cartilage and expanded in monolayer culture before being simultaneously exposed to different concentrations of TXA and VP for varying exposure times. Cell viability and proliferation were assessed using an ATP assay and an Annexin 5 assay, respectively, while changes in the relative expression of chondrogenic marker genes were examined using semiquantitative RT-PCR. Results: The simultaneous exposure of chondrocytes to TXA and VP for more than 48 h led to a reduction in both cell viability and proliferation rates. When exposing chondrocytes to the lowest examined concentrations of both TXA (10 mg/mL) and VP (3 mg/mL), the observed effects were delayed until 96 h. However, our study found no dependencies of the observed effects on the concentrations tested. Further, we found no effects on the expression of chondrogenic marker genes. Conclusions: Consequently, limiting the exposure time of chondrocytes to TXA and VP in an in vitro setting to 24 h may be considered safe and could help to further improve the understanding of the safe use of substances in vivo. However, further in vitro research is required to develop a comprehensive understanding of the effects of both VP and TXA on important periarticular cell types in TKA, including chondrocytes, osteocytes, and tenocytes.

Pharmaceuticals doi: 10.3390/ph17121575

Authors: Othoniel H. Aragon-Martinez Marco M. González-Chávez Othir G. Galicia-Cruz Santiago de J. Méndez-Gallegos Mario A. Isiordia-Espinoza Flavio Martinez-Morales

Background/Objectives: Current urate-lowering therapies may cause serious side effects in patients. Thus, alternative treatments are needed to regulate uric acid (UA) levels in patients with hyperuricemia associated with kidney injury, and natural antioxidant sources have demonstrated utility in this field. For the first time, our study evaluated the effects of an extract of Dactylopius opuntiae insects on the levels of xanthine oxidase (XO) enzymes and synthetic free radicals in vitro and in vivo. Methods: Insects were bred and collected, and two different extracts (D1 and D2) were obtained. For both extracts, XO inhibition and radical scavenging assays were performed. Subsequently, serum purine levels and renal markers were quantified in male BALB/c mice who received a hyperuricemia induction using potassium oxonate, hypoxanthine, and gentamicin. Results: The D2 extract contained 18,037.7 µg/mL of carminic acid, inhibited 53.2% of XO activity at one concentration, and showed IC50 values of 18,207.8 and 5729.6 µg/mL against ABTS and DPPH radicals, respectively. D2 administration reduced serum UA and creatinine levels and prevented an increase in kidney weight and reduction in renal antioxidant capacity caused by hyperuricemia induction and allopurinol use in mice. Despite the satisfactory antioxidant results obtained in vitro, the D1 extract killed the animal models due to its citric acid content. Conclusions: The D2 insect extract can be used as an effective urate-lowering therapy when the increased level of serum uric acid is due to kidney damage.

Pharmaceuticals doi: 10.3390/ph17121574

Authors: Yunxi Ma Yunhan Tao Mingzhu Yuan Xufang Sun

Purpose: Our purpose was to appraise the efficacy and safety of intravitreous vascular endothelial growth factor inhibitor (anti-VEGF) therapy combined with steroids for persistent diabetic macular edema. Methods: A systematic review was conducted of the research evaluating the combination therapy of anti-VEGF and steroids for persistent diabetic macular edema compared to anti-VEGF alone. A meta-analysis was performed using a protocol registered in PROSPERO (CRD42023476333). Continuous and binary variables were extracted. Results were expressed as the mean difference (MD) and risk ratio (RR). Results: A total of 9 trials with 537 eyes were included. The MDs of improvement in best-corrected visual acuity (BCVA) at 1/2/3/6/9/12 months between the combined and monotherapy groups were 1.33 (95% CI [−1.31,3.96]), 3.03 (95% CI [0.01, 6.06]), −0.37 (95% CI [−4.74, 4.00]), −1.37 (95% CI [−4.65, 1.91]), 1.05 (95% CI [−3.68, 5.77]), and 1.70 (95% CI [−3.52, 6.93]). The MDs concerned with a central retinal thickness (CMT) decline in at 1/2/3/6/9/12 months between the two groups were −47.33, 95% CI [−94.35, −0.32]), −89.19 (95% CI [−114.38, −64.00]), −58.84 (95% CI [−96.93, −20.74]), −57.23 (95% CI [−102.62, −11.84]), −40.59 (95% CI [−80.59, −0.58]), and −38.89 (95% CI [−77.38, −0.40]), respectively. Furthermore, the combined group obtained higher relative risks of experiencing events with high intraocular pressure and progressed cataracts. Conclusions: Anti-VEGF combined with ocular steroids showed a significant advantage in improving the retinal anatomical structure compared to anti-VEGF monotherapy for persistent diabetic macular edema. However, as the treatment period extended, the combination treatment was no more effective than monotherapy after 2 months, with more severe side effects.

Pharmaceuticals doi: 10.3390/ph17121573

Authors: Gil Mendes Viana Edézio Ferreira da Cunha-Junior Paloma Wetler Meireles Carreiros Assumpção Marianne Grilo Rezende Yago Sousa dos Santos Emiliano Laiza Maria da Silva Soares Gabriel Rodrigues Coutinho Pereira Carlos Rangel Rodrigues Lucio Mendes Cabral Eduardo Caio Torres-Santos

Background: Leishmaniasis, caused by Leishmania protozoa and transmitted by vectors, presents varied clinical manifestations based on parasite species and host immunity. The lack of effective vaccines or treatments has prompted research into new therapies, including thiourea derivatives, which have demonstrated antiprotozoal activities. Methods: We synthesized two series of N,N′-disubstituted thiourea derivatives through the reaction of isothiocyanates with amines. These compounds were evaluated in vitro against promastigote and amastigote forms of L. amazonensis, alongside cytotoxicity assessments on macrophages. In silico studies were conducted to analyze structure–activity relationships (SARs) and drug-likeness. Results: A total of fifty thiourea derivatives were synthesized and tested. Compound 3e from the first generation exhibited significant anti-leishmanial activity with an IC50 of 4.9 ± 1.2 µM and over 80-fold selectivity compared to that of miltefosine (IC50 = 7.5 ± 1.2 µM). The introduction of a piperazine ring in the second-generation thioureas enhanced potency and selectivity, with compound 5i achieving an IC50 of 1.8 ± 0.5 µM and a selectivity index of approximately 70. Pharmacokinetic predictions indicated favorable profiles for the active compounds. Conclusions: SAR and ADMET analyses identified compound 5i as the most promising candidate for further preclinical evaluation, suggesting that piperazine thiourea derivatives represent a novel class of anti-leishmanial agents.