Pub Date : 2026-01-06DOI: 10.1016/j.ejps.2026.107428
Simona Marzano , Maria Grazia Ferraro , Nicola Grasso , Rossella Buono , Valentina Arciuolo , Federica Iazzetti , Marialuisa Piccolo , Assunta Passarelli , Federica D’Aria , Francesco Merlino , Paolo Grieco , Antonio Randazzo , Bruno Pagano , Carlo Irace , Jussara Amato
AS1411 is a G-rich DNA aptamer that exhibits intrinsic antitumor activity through selective binding to nucleolin, a protein overexpressed in many cancers. Beyond its cytotoxic effects, AS1411 can also serve as an effective targeting ligand for the delivery of therapeutics with poor cellular uptake, including peptide-based drugs. One such candidate is the pro-apoptotic peptide KLA, which selectively disrupts mitochondrial membranes and induces apoptosis upon internalization. In this study, AS1411-KLA conjugates were designed and synthesized using strain-promoted azide-alkyne cycloaddition chemistry, incorporating protease-cleavable peptide linkers to enable intracellular release of both the aptamer and peptide as independent active units. Circular dichroism analysis showed that, in all constructs, the AS1411 domain preserved the characteristic G-quadruplex structural features, while surface plasmon resonance experiments revealed that all conjugates retained nucleolin binding. The biological activity of the conjugates was evaluated in MCF-7 breast cancer cells and in non-tumorigenic MCF-10A cells, and compared with that of the individual aptamer and peptide. One conjugate displayed an enhanced antiproliferative effect compared to the unconjugated components, underscoring the therapeutic potential of this modular design. Overall, this work demonstrates the potential of aptamer-peptide conjugates as a promising strategy for next-generation targeted cancer therapeutics, combining targeted delivery with synergistic therapeutic effects.
{"title":"Design and evaluation of dual-functional aptamer-peptide conjugates as a platform for targeted cancer therapy","authors":"Simona Marzano , Maria Grazia Ferraro , Nicola Grasso , Rossella Buono , Valentina Arciuolo , Federica Iazzetti , Marialuisa Piccolo , Assunta Passarelli , Federica D’Aria , Francesco Merlino , Paolo Grieco , Antonio Randazzo , Bruno Pagano , Carlo Irace , Jussara Amato","doi":"10.1016/j.ejps.2026.107428","DOIUrl":"10.1016/j.ejps.2026.107428","url":null,"abstract":"<div><div>AS1411 is a G-rich DNA aptamer that exhibits intrinsic antitumor activity through selective binding to nucleolin, a protein overexpressed in many cancers. Beyond its cytotoxic effects, AS1411 can also serve as an effective targeting ligand for the delivery of therapeutics with poor cellular uptake, including peptide-based drugs. One such candidate is the pro-apoptotic peptide KLA, which selectively disrupts mitochondrial membranes and induces apoptosis upon internalization. In this study, AS1411-KLA conjugates were designed and synthesized using strain-promoted azide-alkyne cycloaddition chemistry, incorporating protease-cleavable peptide linkers to enable intracellular release of both the aptamer and peptide as independent active units. Circular dichroism analysis showed that, in all constructs, the AS1411 domain preserved the characteristic G-quadruplex structural features, while surface plasmon resonance experiments revealed that all conjugates retained nucleolin binding. The biological activity of the conjugates was evaluated in MCF-7 breast cancer cells and in non-tumorigenic MCF-10A cells, and compared with that of the individual aptamer and peptide. One conjugate displayed an enhanced antiproliferative effect compared to the unconjugated components, underscoring the therapeutic potential of this modular design. Overall, this work demonstrates the potential of aptamer-peptide conjugates as a promising strategy for next-generation targeted cancer therapeutics, combining targeted delivery with synergistic therapeutic effects.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"218 ","pages":"Article 107428"},"PeriodicalIF":4.7,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145932985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-02DOI: 10.1016/j.ejps.2026.107427
Sofie Heylen , Johan Nicolaï , Stijn Van Asten , Katie De Wagter , Andrea Treyer , Jan Snoeys , Raymond Evers , Stephanie Kourula , Pieter Annaert
In vitro to in vivo extrapolation (IVIVE) methods for hepatic clearance (CLH) prediction often underpredict, partly due to reliance on mathematical liver disposition models such as the well-stirred model (WSM) or parallel tube model (PTM). The ex vivo isolated perfused rat liver (IPRL) model bridges in vitro and in vivo data, providing mechanistic insights into the predictive accuracy of IVIVE models. This study evaluates the IPRL model across a diverse selection of 16 compounds, and benchmarks results against in vitro and in vivo data to verify the predictive performance of the WSM and PTM. Results demonstrate that both the IPRL and in vivo clearance conflict with assumptions of the WSM (AAFE = 2.85) or PTM (AAFE = 1.74), which consider the liver outlet concentration as a driver for the hepatic elimination rate. However, except for terfenadine, IPRL clearance predictions were within two-fold (AAFE = 1.59) of in vivo clearance when the liver inlet concentration was utilized to calculate the CLH. When employing the WSM or PTM for in vitro to ex vivo extrapolation, underpredictions were observed for compounds with high plasma protein binding and subject to sinusoidal hepatic uptake, reflecting model oversimplification compared to in vivo dynamics. Our findings experimentally challenge the theoretical assumptions underlying the use of the WSM and PTM in IVIVE methods. Unique insights from the IPRL model point to the next steps needed to advance IVIVE: refining current liver disposition models through enhanced and next-generation in vitro assays, capturing dynamic in vivo disposition mechanisms, and exploring complementary models.
{"title":"Deconvoluting the in vitro to in vivo drug clearance gap: Questioning the predictive performance of traditional hepatic clearance models","authors":"Sofie Heylen , Johan Nicolaï , Stijn Van Asten , Katie De Wagter , Andrea Treyer , Jan Snoeys , Raymond Evers , Stephanie Kourula , Pieter Annaert","doi":"10.1016/j.ejps.2026.107427","DOIUrl":"10.1016/j.ejps.2026.107427","url":null,"abstract":"<div><div><em>In vitro</em> to <em>in vivo</em> extrapolation (IVIVE) methods for hepatic clearance (CL<sub>H</sub>) prediction often underpredict, partly due to reliance on mathematical liver disposition models such as the well-stirred model (WSM) or parallel tube model (PTM). The <em>ex vivo</em> isolated perfused rat liver (IPRL) model bridges <em>in vitro</em> and <em>in vivo</em> data, providing mechanistic insights into the predictive accuracy of IVIVE models. This study evaluates the IPRL model across a diverse selection of 16 compounds, and benchmarks results against <em>in vitro</em> and <em>in vivo</em> data to verify the predictive performance of the WSM and PTM. Results demonstrate that both the IPRL and <em>in vivo</em> clearance conflict with assumptions of the WSM (AAFE = 2.85) or PTM (AAFE = 1.74), which consider the liver outlet concentration as a driver for the hepatic elimination rate. However, except for terfenadine, IPRL clearance predictions were within two-fold (AAFE = 1.59) of <em>in vivo</em> clearance when the liver inlet concentration was utilized to calculate the CL<sub>H</sub>. When employing the WSM or PTM for <em>in vitro</em> to <em>ex vivo</em> extrapolation, underpredictions were observed for compounds with high plasma protein binding and subject to sinusoidal hepatic uptake, reflecting model oversimplification compared to <em>in vivo</em> dynamics. Our findings experimentally challenge the theoretical assumptions underlying the use of the WSM and PTM in IVIVE methods. Unique insights from the IPRL model point to the next steps needed to advance IVIVE: refining current liver disposition models through enhanced and next-generation <em>in vitro</em> assays, capturing dynamic <em>in vivo</em> disposition mechanisms, and exploring complementary models.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"218 ","pages":"Article 107427"},"PeriodicalIF":4.7,"publicationDate":"2026-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145899620","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The purpose of this study was to evaluate how the dissolution-supersaturation-precipitation (DSP) behaviour of cocrystals translates into drug permeation, employing the biomimetic PermeaPad® barrier in a side-by-side cell set-up. Building on our previous work, which demonstrated that reducing an unnecessarily high cocrystal solubility advantage (SA = Scocrystal/Sdrug) through the generation of non-stoichiometric solution conditions can significantly extend the drug supersaturation, we now investigate this approach in the presence of an absorptive environment. Our findings with the 1:1 ketoconazole (KTZ) -p-aminobenzoic acid (PABA) cocrystal indicate the existence of an optimal SA value (7), where KTZ-PABA dissolution performance resulted in 8-fold AUC increase, translating to a 7-fold increase in cumulative KTZ permeation after 6 h. This enhancement was achieved by dissolving cocrystal (based on the drug therapeutic dose) with an additional coformer solid phase, rationally designed using a graphical approach grounded in the cocrystal thermodynamic (Ksp, Scocrystal, Sdrug) and kinetic parameters (σcrit). The strategy presented in this work can be readily applied during the pre-formulation stage of cocrystal development, enabling targeted selection of coformer concentrations for formulation development. Overall cocrystals offer unique flexibility as a formulation strategy for poorly water-soluble drugs, allowing for tailored DSP behaviour and drug transport over biological barriers based on mechanistic understanding, as demonstrated in this study.
{"title":"Impact of cocrystal dissolution-supersaturation-precipitation (DSP) behaviour on drug permeation across the PermeaPad® biomimetic barrier","authors":"Lucy May Newman , Matteo Guidetti , Annette Bauer-Brandl , Naír Rodríguez-Hornedo , Tatiane Cogo Machado","doi":"10.1016/j.ejps.2025.107426","DOIUrl":"10.1016/j.ejps.2025.107426","url":null,"abstract":"<div><div>The purpose of this study was to evaluate how the dissolution-supersaturation-precipitation (DSP) behaviour of cocrystals translates into drug permeation<strong>,</strong> employing the biomimetic PermeaPad® barrier in a side-by-side cell set-up. Building on our previous work, which demonstrated that reducing an unnecessarily high cocrystal solubility advantage (SA = S<sub>cocrystal</sub>/S<sub>drug</sub>) through the generation of non-stoichiometric solution conditions can significantly extend the drug supersaturation, we now investigate this approach in the presence of an absorptive environment. Our findings with the 1:1 ketoconazole (KTZ) -<em>p</em>-aminobenzoic acid (PABA) cocrystal indicate the existence of an optimal SA value (7), where KTZ-PABA dissolution performance resulted in 8-fold AUC increase, translating to a 7-fold increase in cumulative KTZ permeation after 6 h. This enhancement was achieved by dissolving cocrystal (based on the drug therapeutic dose) with an additional coformer solid phase, rationally designed using a graphical approach grounded in the cocrystal thermodynamic (K<sub>sp</sub>, S<sub>cocrystal</sub>, S<sub>drug</sub>) and kinetic parameters (σ<sub>crit</sub>). The strategy presented in this work can be readily applied during the pre-formulation stage of cocrystal development, enabling targeted selection of coformer concentrations for formulation development. Overall cocrystals offer unique flexibility as a formulation strategy for poorly water-soluble drugs, allowing for tailored DSP behaviour and drug transport over biological barriers based on mechanistic understanding<strong>,</strong> as demonstrated in this study.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"218 ","pages":"Article 107426"},"PeriodicalIF":4.7,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145896350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-31DOI: 10.1016/j.ejps.2025.107425
Venkata Krishna Rao Balaga, Argyro Chatziadi, Luděk Ridvan, Miroslav Šoóš
Coamorphization is an attractive approach to modifying the physicochemical properties of drug molecules, especially the solubility, dissolution, and associated bioavailability. Although these formulations may be advantageous, they exhibit poor physical stability and undergo recrystallisation. To address this limitation, this study investigates the effect of positional isomerism on the coamorphous formation and associated physicochemical properties, to select an optimum solid form with improved stability. Enzalutamide (ENZ), a BCS class II drug, was used as a model compound. Four positional isomers including 2,3-, 2,4-, 2,5- and 2,6-dihydroxybenzoic acid (DHB) were used as coamorphous coformers. Coamorphous formulations were prepared by ball mill in a 2:1 molecular ratio (API:coformer). The solid-state properties of the prepared coamorphous forms were characterised using X-ray powder diffractometer (XRPD), modulated differential scanning calorimetry (mDSC), and Fourier transformed infrared spectrometry (FTIR). Additionally, intra isomer variability in the amorphization kinetics and dissolution enhancement of ENZ, along with physical stability, were evaluated. All coformers formed coamorphous systems, as confirmed by XRPD. mDSC data showed that the glass transition temperature (Tg) varied among the prepared coamorphous forms and was lower than that of pure ENZ. Although there was no significant difference in the dissolution behaviour, the physical stability data reveal a contrast trend. Among the prepared coamorphous forms, ENZ-24DHBCAM exhibited superior stability, while ENZ-26DHBCAM exhibited poor stability. This article summarises the similarities and differences between the physicochemical properties of coamorphous forms of Enz because of the change in coformer positional isomerism. Stability studies under different humidity conditions revealed significant differences: at 40% RH, all coamorphous forms remained stable for up to 8 weeks, with minor deviations for ENZ-23DHBCAM and ENZ-25DHBCAM. Under 75% RH, the stability varied markedly; ENZ-24DHBCAM maintained stability for at least 8 weeks, while ENZ-26DHBCAM became unstable within 1-2 weeks, and ENZ-AMP and ENZ-23DHBCAM lost stability by week 6. These results demonstrate the careful selection of coformer positional isomer can quantitatively enhance the stability of coamorphous forms, highlighting the importance of positional isomerism associated chemical design space in optimizing solid-state properties.
{"title":"Structurally Similar, Functionally Different: Impact of Coformer Positional Isomerism on Co-Amorphous Enzalutamide.","authors":"Venkata Krishna Rao Balaga, Argyro Chatziadi, Luděk Ridvan, Miroslav Šoóš","doi":"10.1016/j.ejps.2025.107425","DOIUrl":"https://doi.org/10.1016/j.ejps.2025.107425","url":null,"abstract":"<p><p>Coamorphization is an attractive approach to modifying the physicochemical properties of drug molecules, especially the solubility, dissolution, and associated bioavailability. Although these formulations may be advantageous, they exhibit poor physical stability and undergo recrystallisation. To address this limitation, this study investigates the effect of positional isomerism on the coamorphous formation and associated physicochemical properties, to select an optimum solid form with improved stability. Enzalutamide (ENZ), a BCS class II drug, was used as a model compound. Four positional isomers including 2,3-, 2,4-, 2,5- and 2,6-dihydroxybenzoic acid (DHB) were used as coamorphous coformers. Coamorphous formulations were prepared by ball mill in a 2:1 molecular ratio (API:coformer). The solid-state properties of the prepared coamorphous forms were characterised using X-ray powder diffractometer (XRPD), modulated differential scanning calorimetry (mDSC), and Fourier transformed infrared spectrometry (FTIR). Additionally, intra isomer variability in the amorphization kinetics and dissolution enhancement of ENZ, along with physical stability, were evaluated. All coformers formed coamorphous systems, as confirmed by XRPD. mDSC data showed that the glass transition temperature (Tg) varied among the prepared coamorphous forms and was lower than that of pure ENZ. Although there was no significant difference in the dissolution behaviour, the physical stability data reveal a contrast trend. Among the prepared coamorphous forms, ENZ-24DHB<sup>CAM</sup> exhibited superior stability, while ENZ-26DHB<sup>CAM</sup> exhibited poor stability. This article summarises the similarities and differences between the physicochemical properties of coamorphous forms of Enz because of the change in coformer positional isomerism. Stability studies under different humidity conditions revealed significant differences: at 40% RH, all coamorphous forms remained stable for up to 8 weeks, with minor deviations for ENZ-23DHB<sup>CAM</sup> and ENZ-25DHB<sup>CAM</sup>. Under 75% RH, the stability varied markedly; ENZ-24DHB<sup>CAM</sup> maintained stability for at least 8 weeks, while ENZ-26DHB<sup>CAM</sup> became unstable within 1-2 weeks, and ENZ-AMP and ENZ-23DHB<sup>CAM</sup> lost stability by week 6. These results demonstrate the careful selection of coformer positional isomer can quantitatively enhance the stability of coamorphous forms, highlighting the importance of positional isomerism associated chemical design space in optimizing solid-state properties.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"107425"},"PeriodicalIF":4.7,"publicationDate":"2025-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145892224","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-28DOI: 10.1016/j.ejps.2025.107424
Praveen Kumar Singh , Matej Zore , Paola San-Martin-Galindo , Moritz M. Kornmayer , Inés Reigada , Leena Hanski , Kirsi Savijoki , Jari Yli-Kauhaluoma , Jayendra Z. Patel
Febuxostat, a marketed anti-gout drug, has been reported to inhibit quorum sensing (QS), biofilm formation, and virulence in Gram-negative bacteria Pseudomonas aeruginosa PAO1 and Chromobacterium violaceum CV026. However, no structure-activity relationship (SAR) studies have been reported to date. To address this gap, we synthesized 27 novel febuxostat analogs and evaluated their QS inhibitory activity using wild-type C. violaceum and the mutant strain CV026. This dual-strain assay enabled differentiation between quorum sensing inhibition and quorum quenching mechanisms, allowing determination of whether QS interference occurred at the level of signal synthesis or signal reception.
Here, we demonstrated that febuxostat acts as a quorum quencher at concentrations lower than previously reported, strongly inhibiting violacein production in CV026 without affecting bacterial viability. At 400 µM, several analogs showed comparable or stronger activity than febuxostat in one or both strains. At 40 µM, trifluoromethoxyphenyl analog 49 emerged as the most effective QS inhibitor in the wild-type strain, whereas cyclopentyl analog 24 was most active in CV026, with a few other derivatives also retaining notable activity. We also assessed whether febuxostat and its analogs exhibit antibacterial activity. While febuxostat showed no antibacterial activity, seven analogs (26–30, 33, and 49) exhibited minimum inhibitory concentrations (MICs) ranging from 2.5 to 40 µM against Gram-positive bacteria Staphylococcus aureus, S. epidermidis, Enterococcus faecalis, and E. faecium. On the other hand, no activity was observed against Gram-negative bacteria. In follow-up studies, the two most potent antibacterial compounds 29 (MIC = 5 µM) and 30 (MIC = 10 µM), showed no cytotoxicity against the mammalian cell line A549, while they were less tolerated by THP-1 cells. In summary, our results provide SAR insights into febuxostat analogs and highlight their potential as novel anti-QS and antibacterial agents against multidrug-resistant bacteria.
{"title":"Febuxostat analogs as anti-quorum sensing and antibacterial agents","authors":"Praveen Kumar Singh , Matej Zore , Paola San-Martin-Galindo , Moritz M. Kornmayer , Inés Reigada , Leena Hanski , Kirsi Savijoki , Jari Yli-Kauhaluoma , Jayendra Z. Patel","doi":"10.1016/j.ejps.2025.107424","DOIUrl":"10.1016/j.ejps.2025.107424","url":null,"abstract":"<div><div>Febuxostat, a marketed anti-gout drug, has been reported to inhibit quorum sensing (QS), biofilm formation, and virulence in Gram-negative bacteria <em>Pseudomonas aeruginos</em>a PAO1 and <em>Chromobacterium violaceum</em> CV026. However, no structure-activity relationship (SAR) studies have been reported to date. To address this gap, we synthesized 27 novel febuxostat analogs and evaluated their QS inhibitory activity using wild-type <em>C. violaceum</em> and the mutant strain CV026. This dual-strain assay enabled differentiation between quorum sensing inhibition and quorum quenching mechanisms, allowing determination of whether QS interference occurred at the level of signal synthesis or signal reception.</div><div>Here, we demonstrated that febuxostat acts as a quorum quencher at concentrations lower than previously reported, strongly inhibiting violacein production in CV026 without affecting bacterial viability. At 400 µM, several analogs showed comparable or stronger activity than febuxostat in one or both strains. At 40 µM, trifluoromethoxyphenyl analog <strong>49</strong> emerged as the most effective QS inhibitor in the wild-type strain, whereas cyclopentyl analog <strong>24</strong> was most active in CV026, with a few other derivatives also retaining notable activity. We also assessed whether febuxostat and its analogs exhibit antibacterial activity. While febuxostat showed no antibacterial activity, seven analogs (<strong>26</strong>–<strong>30, 33</strong>, and <strong>49</strong>) exhibited minimum inhibitory concentrations (MICs) ranging from 2.5 to 40 µM against Gram-positive bacteria <em>Staphylococcus aureus, S. epidermidis, Enterococcus faecalis</em>, and <em>E. faecium</em>. On the other hand, no activity was observed against Gram-negative bacteria. In follow-up studies, the two most potent antibacterial compounds <strong>29</strong> (MIC = 5 µM) and <strong>30</strong> (MIC = 10 µM), showed no cytotoxicity against the mammalian cell line A549, while they were less tolerated by THP-1 cells. In summary, our results provide SAR insights into febuxostat analogs and highlight their potential as novel anti-QS and antibacterial agents against multidrug-resistant bacteria.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"218 ","pages":"Article 107424"},"PeriodicalIF":4.7,"publicationDate":"2025-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145862528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-22DOI: 10.1016/j.ejps.2025.107423
Shengjie Cong , Yonggang Yang , Rasoul Akram
Bone tissue engineering (BTE) is an attempt to overcome the limitations of conventional grafting through the combination of bioactive scaffolds and regenerative signals. In this study, we prepared an extrusion-based 3D-printed composite scaffold composed of chitosan (CH) combined with dexamethasone (DEX)-loaded mesoporous silica nanoparticle (MSN), (MSN@DEX) in order to compensate for both mechanical insufficiency and temporary osteoinductive signal. MSNs were synthesized by a templated sol-gel method with high drug loading efficiency and biphasic release behavior. Inclusion of MSN@DEX in chitosan resulted in scaffolds with homogeneous and interconnected porosity (310–420 μm), high compressive strength, good swelling profile, and a degradation profile consistent with the time course of bone healing. In vitro experiments with mesenchymal stem cells (MSCs) resulted in a high hemocompatibility, a sustained cell proliferation and a significantly enhanced osteogenic differentiation given by alkaline phosphatase (ALP) activity, calcium deposits and stage-dependent upregulation of RUNX2, ALP, COL1A1 and OCN genes and protein. Together, the hierarchical scaffold architecture, nanostructured reinforcement, and local sustained DEX release provide a cost-effective, stable and clinically adaptable platform for robust osteogenesis with the possibility of enhanced osteogenic induction even with lower dexamethasone release. These findings underscore the potential of CH‑MSN@DEX scaffolds for bone regeneration applications. By combining structural reinforcement with sustained osteoinductive signaling within a single printable construct, this approach represents a clear advancement over previously reported chitosan‑based or DEX‑releasing scaffold systems.
{"title":"3-D printing of chitosan-MSN@dexamethasone scaffold with enhanced properties for bone tissue engineering","authors":"Shengjie Cong , Yonggang Yang , Rasoul Akram","doi":"10.1016/j.ejps.2025.107423","DOIUrl":"10.1016/j.ejps.2025.107423","url":null,"abstract":"<div><div>Bone tissue engineering (BTE) is an attempt to overcome the limitations of conventional grafting through the combination of bioactive scaffolds and regenerative signals. In this study, we prepared an extrusion-based 3D-printed composite scaffold composed of chitosan (CH) combined with dexamethasone (DEX)-loaded mesoporous silica nanoparticle (MSN), (MSN@DEX) in order to compensate for both mechanical insufficiency and temporary osteoinductive signal. MSNs were synthesized by a templated sol-gel method with high drug loading efficiency and biphasic release behavior. Inclusion of MSN@DEX in chitosan resulted in scaffolds with homogeneous and interconnected porosity (310–420 μm), high compressive strength, good swelling profile, and a degradation profile consistent with the time course of bone healing. In vitro experiments with mesenchymal stem cells (MSCs) resulted in a high hemocompatibility, a sustained cell proliferation and a significantly enhanced osteogenic differentiation given by alkaline phosphatase (ALP) activity, calcium deposits and stage-dependent upregulation of RUNX2, ALP, COL1A1 and OCN genes and protein. Together, the hierarchical scaffold architecture, nanostructured reinforcement, and local sustained DEX release provide a cost-effective, stable and clinically adaptable platform for robust osteogenesis with the possibility of enhanced osteogenic induction even with lower dexamethasone release. These findings underscore the potential of CH‑MSN@DEX scaffolds for bone regeneration applications. By combining structural reinforcement with sustained osteoinductive signaling within a single printable construct, this approach represents a clear advancement over previously reported chitosan‑based or DEX‑releasing scaffold systems.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"218 ","pages":"Article 107423"},"PeriodicalIF":4.7,"publicationDate":"2025-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145827119","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-21DOI: 10.1016/j.ejps.2025.107422
Valerio Ciccone , Claudia Cecchin , Maria Frosini , Mario Saletti , Samuele Maramai , Germano Giuliani , Marco Paolino , Andrea Cappelli , Maurizio Anzini , Sandra Donnini , Lucia Morbidelli
Cyclooxygenase-2 (COX-2) is overexpressed in various cancers and has emerged as a promising target in oncological pharmacotherapy.
This study investigates the in vitro antitumor properties and mechanism of action of novel vicinal diaryl-substituted heterocyclic COX-2 inhibitors, with a focus on VA1213, in comparison to celecoxib, a widely marketed COX-2 inhibitor known for its off-target effects. We assessed cytotoxicity, apoptosis induction, cell-cycle distribution, antimetastatic activity, and alterations in key signaling pathways in HT-29 colorectal carcinoma and MDA-MB-231 breast carcinoma cell lines. Among the novel compounds, VA1213 exhibited the most potent growth-inhibitory activity, demonstrating time-dependent cytotoxicity with a lower IC50 after 48–72 h of treatment compared to VA692 and VA694, and consistent with that observed for celecoxib.
Unlike celecoxib, which produced rapid cytotoxic effects, VA1213 required prolonged exposure, suggesting a distinct mechanism of action. VA1213 induced G₀/G₁ phase cell cycle arrest and apoptosis via caspase-3 activation. Furthermore, it impaired EGFR downstream signaling by reducing ERK1/2 and AKT phosphorylation, without directly inhibiting EGFR itself. At sub-cytotoxic concentrations, VA1213 was more effective than celecoxib in inhibiting cell migration and demonstrated a comparable reduction in clonogenic potential. These findings highlight VA1213 as a COX-2 inhibitor with noteworthy in vitro antitumor efficacy, comparable to that of celecoxib. Its ability to interfere with multiple cancer-associated signaling pathways and reduce tumor cell aggressiveness underscores its potential as a promising therapeutic candidate. Further in vivo studies are warranted to confirm its efficacy and assess potential off-target effects.
{"title":"VA1213, a selective COX-2 inhibitor, exhibits antitumor activity by suppressing EGFR, AKT, and ERK1/2 phosphorylation","authors":"Valerio Ciccone , Claudia Cecchin , Maria Frosini , Mario Saletti , Samuele Maramai , Germano Giuliani , Marco Paolino , Andrea Cappelli , Maurizio Anzini , Sandra Donnini , Lucia Morbidelli","doi":"10.1016/j.ejps.2025.107422","DOIUrl":"10.1016/j.ejps.2025.107422","url":null,"abstract":"<div><div>Cyclooxygenase-2 (COX-2) is overexpressed in various cancers and has emerged as a promising target in oncological pharmacotherapy.</div><div>This study investigates the in vitro antitumor properties and mechanism of action of novel vicinal diaryl-substituted heterocyclic COX-2 inhibitors, with a focus on <strong>VA1213</strong>, in comparison to celecoxib, a widely marketed COX-2 inhibitor known for its off-target effects. We assessed cytotoxicity, apoptosis induction, cell-cycle distribution, antimetastatic activity, and alterations in key signaling pathways in HT-29 colorectal carcinoma and MDA-MB-231 breast carcinoma cell lines. Among the novel compounds, <strong>VA1213</strong> exhibited the most potent growth-inhibitory activity, demonstrating time-dependent cytotoxicity with a lower IC<sub>50</sub> after 48–72 h of treatment compared to <strong>VA692</strong> and <strong>VA694</strong>, and consistent with that observed for celecoxib.</div><div>Unlike celecoxib, which produced rapid cytotoxic effects, <strong>VA1213</strong> required prolonged exposure, suggesting a distinct mechanism of action. <strong>VA1213</strong> induced G₀/G₁ phase cell cycle arrest and apoptosis via caspase-3 activation. Furthermore, it impaired EGFR downstream signaling by reducing ERK1/2 and AKT phosphorylation, without directly inhibiting EGFR itself. At sub-cytotoxic concentrations, <strong>VA1213</strong> was more effective than celecoxib in inhibiting cell migration and demonstrated a comparable reduction in clonogenic potential. These findings highlight <strong>VA1213</strong> as a COX-2 inhibitor with noteworthy in vitro antitumor efficacy, comparable to that of celecoxib. Its ability to interfere with multiple cancer-associated signaling pathways and reduce tumor cell aggressiveness underscores its potential as a promising therapeutic candidate. Further in vivo studies are warranted to confirm its efficacy and assess potential off-target effects.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"219 ","pages":"Article 107422"},"PeriodicalIF":4.7,"publicationDate":"2025-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145818640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-18DOI: 10.1016/j.ejps.2025.107412
Conglu Zhao , Liyuan Ji , Xiaoting Wang , Jia Zhang , Xiang Xu , Xiaoting Zhang , Yanjie Ding , Keran Li , Chaoyue Zheng , Kaijun Qiu , Jing Yan , Songtao Gu , Honggang Zhou , Cheng Yang , Hongli Li , Xiaoting Gu , Xiaoyu Ai
{"title":"Erratum to “Intratracheal instillation of chitosan-coated formononetin-loaded porous microspheres prolongs lung retention and improves the treatment” [European Journal of Pharmaceutical Sciences. 2025 Dec 1;215:107347]","authors":"Conglu Zhao , Liyuan Ji , Xiaoting Wang , Jia Zhang , Xiang Xu , Xiaoting Zhang , Yanjie Ding , Keran Li , Chaoyue Zheng , Kaijun Qiu , Jing Yan , Songtao Gu , Honggang Zhou , Cheng Yang , Hongli Li , Xiaoting Gu , Xiaoyu Ai","doi":"10.1016/j.ejps.2025.107412","DOIUrl":"10.1016/j.ejps.2025.107412","url":null,"abstract":"","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107412"},"PeriodicalIF":4.7,"publicationDate":"2025-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145793541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-17DOI: 10.1016/j.ejps.2025.107417
Wen‐Chien Huang , Chieh-Yung Wang , Ying Chen , Yi-Hsuan Lin , Chih-Ying Changchien , Chih-Feng Chian , Chen-Liang Tsai
Background
Non-small cell lung cancer (NSCLC) frequently develops resistance to EGFR-tyrosine kinase inhibitors (TKIs), particularly in malignant pleural effusion (MPE), where a highly acidic tumor microenvironment promotes the emergence of drug-tolerant persister (DTP) cells. Building upon previous evidence that MPE acidity drives metabolic plasticity, this study investigates how acid-sensing ion channel 3 (ASIC3) coordinates PLCG1-dependent macropinocytosis to maintain osimertinib tolerance.
Methods
Primary MPE-derived NSCLC cultures underwent next-generation sequencing to define mutational heterogeneity. Acidic (pH 6.5–6.8) conditions were used to generate osimertinib-induced DTP (Osi-DTP) models. Functional assays assessed viability, invasion, colony formation, autophagy, and macropinocytosis. ASIC3 was silenced using shRNA, followed by Seahorse metabolic analysis. Transcriptomic profiling identified differentially expressed genes. ASIC3-targeted therapeutic interventions were evaluated in vitro and in xenograft models.
Results
MPE-derived NSCLC cultures showed substantial genomic diversity, including EGFR exon 19 deletion and T790M mutations. Acid-adapted Osi-DTP cells exhibited EMT-like phenotypes, reduced proliferation, elevated stemness markers, and strong activation of stress-response pathways. ASIC3 was consistently upregulated under acidic conditions and drove PLCG1-mediated macropinocytosis to support nutrient scavenging and survival. ASIC3 knockdown markedly reduced DTP cell viability, invasiveness, colony formation, autophagy, and glycolysis, while inducing a metabolic shift toward oxidative phosphorylation. In vivo, combining an ASIC3 inhibitor with osimertinib significantly delayed tumor progression and improved survival without added toxicity. Clinically, high ASIC3 expression correlated with poor overall survival and increased autophagy-associated markers.
Conclusion
ASIC3 is a central regulator of acidosis-driven drug persistence in MPE-associated NSCLC, sustaining osimertinib tolerance through PLCG1-dependent macropinocytosis and metabolic reprogramming. Targeting ASIC3 restores TKI sensitivity by disrupting nutrient acquisition, autophagy, and metabolic adaptation, representing a promising therapeutic strategy to prevent or delay EGFR-TKI resistance.
{"title":"ASIC3–PLCG1 axis–driven macropinocytosis promotes osimertinib drug-tolerant persistence in malignant pleural effusion–associated NSCLC","authors":"Wen‐Chien Huang , Chieh-Yung Wang , Ying Chen , Yi-Hsuan Lin , Chih-Ying Changchien , Chih-Feng Chian , Chen-Liang Tsai","doi":"10.1016/j.ejps.2025.107417","DOIUrl":"10.1016/j.ejps.2025.107417","url":null,"abstract":"<div><h3>Background</h3><div>Non-small cell lung cancer (NSCLC) frequently develops resistance to EGFR-tyrosine kinase inhibitors (TKIs), particularly in malignant pleural effusion (MPE), where a highly acidic tumor microenvironment promotes the emergence of drug-tolerant persister (DTP) cells. Building upon previous evidence that MPE acidity drives metabolic plasticity, this study investigates how acid-sensing ion channel 3 (ASIC3) coordinates PLCG1-dependent macropinocytosis to maintain osimertinib tolerance.</div></div><div><h3>Methods</h3><div>Primary MPE-derived NSCLC cultures underwent next-generation sequencing to define mutational heterogeneity. Acidic (pH 6.5–6.8) conditions were used to generate osimertinib-induced DTP (Osi-DTP) models. Functional assays assessed viability, invasion, colony formation, autophagy, and macropinocytosis. ASIC3 was silenced using shRNA, followed by Seahorse metabolic analysis. Transcriptomic profiling identified differentially expressed genes. ASIC3-targeted therapeutic interventions were evaluated <em>in vitro</em> and in xenograft models.</div></div><div><h3>Results</h3><div>MPE-derived NSCLC cultures showed substantial genomic diversity, including EGFR exon 19 deletion and T790M mutations. Acid-adapted Osi-DTP cells exhibited EMT-like phenotypes, reduced proliferation, elevated stemness markers, and strong activation of stress-response pathways. ASIC3 was consistently upregulated under acidic conditions and drove PLCG1-mediated macropinocytosis to support nutrient scavenging and survival. ASIC3 knockdown markedly reduced DTP cell viability, invasiveness, colony formation, autophagy, and glycolysis, while inducing a metabolic shift toward oxidative phosphorylation. In vivo, combining an ASIC3 inhibitor with osimertinib significantly delayed tumor progression and improved survival without added toxicity. Clinically, high ASIC3 expression correlated with poor overall survival and increased autophagy-associated markers.</div></div><div><h3>Conclusion</h3><div>ASIC3 is a central regulator of acidosis-driven drug persistence in MPE-associated NSCLC, sustaining osimertinib tolerance through PLCG1-dependent macropinocytosis and metabolic reprogramming. Targeting ASIC3 restores TKI sensitivity by disrupting nutrient acquisition, autophagy, and metabolic adaptation, representing a promising therapeutic strategy to prevent or delay EGFR-TKI resistance.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107417"},"PeriodicalIF":4.7,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145793577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-17DOI: 10.1016/j.ejps.2025.107418
Manxiang Wang , Umara Arif , Tayyaba Kaleem , Humaira Majeed Khan , Abid Mehmood Yousaf , Nadeem Ahmad , Imran Nazir , Yasser Shahzad
The present study is aimed at developing baricitinib (BAR) and tenoxicam (TNX) loaded nanosponge (NS) gel for potential topical treatment of rheumatoid arthritis (RA). DPC crosslinked β-CD NS were synthesized at various molar ratios (1:2–1:20), with the 1:8 ratio showing the highest drug loading efficiency (67.0 ± 6.9% for BAR; 82.1 ± 6.8% for TNX). XRD and DSC analysis confirmed amorphization of drugs, SEM revealed porous morphology, DLS revealed particle sizes of 545.5 ± 16.4 nm (BAR-NS) and 585.2 ± 57.7 nm (TNX-NS) with zeta potentials of −11.1 ± 4.4 and −14.6 ± 3.9 mV, respectively. The optimized NS were formulated into topical gels, demonstrating sustained release (BAR: 77–90%; TNX: 96% within 12 h). In vivo evaluation using a CFA-induced RA rat model revealed that combined BAR-NS and TNX-NS gel (0.05/0.25% w/w) inhibited paw edema by >90% at day 25, significantly downregulated pro-inflammatory cytokines (IL-6, IL-1β, TNF-α), restored IL-10, and normalized liver enzymes (ALT, AST, ALP). Histopathological analysis confirmed preservation of cartilage and bone with minimal synovial infiltration. In conclusion, combination of BAR-NS and TNX-NS gel showed synergistic anti-arthritic and anti-inflammatory effects, thus offering a potential topical alternative to conventional therapies for the management of RA.
{"title":"Diphenyl carbonate crosslinked β-cyclodextrin nanosponge-based topically applied gel for co-delivery of baricitinib and tenoxicam in rheumatoid arthritis","authors":"Manxiang Wang , Umara Arif , Tayyaba Kaleem , Humaira Majeed Khan , Abid Mehmood Yousaf , Nadeem Ahmad , Imran Nazir , Yasser Shahzad","doi":"10.1016/j.ejps.2025.107418","DOIUrl":"10.1016/j.ejps.2025.107418","url":null,"abstract":"<div><div>The present study is aimed at developing baricitinib (BAR) and tenoxicam (TNX) loaded nanosponge (NS) gel for potential topical treatment of rheumatoid arthritis (RA). DPC crosslinked β-CD NS were synthesized at various molar ratios (1:2–1:20), with the 1:8 ratio showing the highest drug loading efficiency (67.0 ± 6.9% for BAR; 82.1 ± 6.8% for TNX). XRD and DSC analysis confirmed amorphization of drugs, SEM revealed porous morphology, DLS revealed particle sizes of 545.5 ± 16.4 nm (BAR-NS) and 585.2 ± 57.7 nm (TNX-NS) with zeta potentials of −11.1 ± 4.4 and −14.6 ± 3.9 mV, respectively. The optimized NS were formulated into topical gels, demonstrating sustained release (BAR: 77–90%; TNX: 96% within 12 h). In vivo evaluation using a CFA-induced RA rat model revealed that combined BAR-NS and TNX-NS gel (0.05/0.25% w/w) inhibited paw edema by >90% at day 25, significantly downregulated pro-inflammatory cytokines (IL-6, IL-1β, TNF-α), restored IL-10, and normalized liver enzymes (ALT, AST, ALP). Histopathological analysis confirmed preservation of cartilage and bone with minimal synovial infiltration. In conclusion, combination of BAR-NS and TNX-NS gel showed synergistic anti-arthritic and anti-inflammatory effects, thus offering a potential topical alternative to conventional therapies for the management of RA.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107418"},"PeriodicalIF":4.7,"publicationDate":"2025-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145793517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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