Pub Date : 2025-02-01DOI: 10.1016/j.ejps.2024.106986
Joana F.P.R. Terreiro, Joaquim T. Marquês, Inês Antunes, Catarina Frazão de Faria, Susana Santos, Filomena Martins, Rodrigo F.M. de Almeida
Tuberculosis is one of the leading causes of mortality worldwide due to the growth of multi-drug resistant strains unsusceptible to currently available therapies. Four compounds, isoniazid (INH) and three derivatives, N'-decanoylisonicotinohydrazide (INHC10), N'-(E)-(4-phenoxybenzylidene)isonicotinohydrazide (N34) and N’-(4-phenoxybenzyl)isonicotinohydrazide (N34red), were studied. Owing to their advantageous in vitro selectivity index against the primary mutation responsible for drug resistance in Mycobacterium tuberculosis (Mtb), as well as their suitable lipophilicity and interaction with human serum albumin, INHC10 and N34 were deemed promising antitubercular compounds. N34red, despite differing from N34 only in the saturation of the N′ = C bond, presents a poor selectivity index. To delve deeper into the therapeutic potential of these compounds, their interaction with biomembrane models, mimicking biological barriers on the way to the target inside Mtb cells, was herein evaluated. All compounds, except N34red, weakened the packing of the acyl chains in the rigid lipid gel phase, especially INHC10, which was the only compound disturbing liquid disordered membranes. Notably, all compounds except INH decreased membrane dipole potential, across all types of bilayers studied, but only N34red had a drastic effect. The insertion in gel phase bilayers suggests that the compounds may be able to penetrate the rigid cell wall of Mtb. Förster's resonance energy transfer (FRET) assays in ternary bilayers with liquid ordered/liquid disordered lateral heterogeneity mimicking human cell membranes, showed that the compounds affected neither the size nor the organization of lipid domains. These results provide molecular insights into the low toxicity against human cell lines and improved activity against drug-resistant Mtb of INHC10 and N34.
{"title":"Membrane interaction studies of isoniazid derivatives active against drug-resistant tuberculosis","authors":"Joana F.P.R. Terreiro, Joaquim T. Marquês, Inês Antunes, Catarina Frazão de Faria, Susana Santos, Filomena Martins, Rodrigo F.M. de Almeida","doi":"10.1016/j.ejps.2024.106986","DOIUrl":"10.1016/j.ejps.2024.106986","url":null,"abstract":"<div><div>Tuberculosis is one of the leading causes of mortality worldwide due to the growth of multi-drug resistant strains unsusceptible to currently available therapies. Four compounds, isoniazid (INH) and three derivatives, <em>N</em>'-decanoylisonicotinohydrazide (INH<img>C10), <em>N</em>'-(<em>E</em>)-(4-phenoxybenzylidene)isonicotinohydrazide (N34) and <em>N</em>’-(4-phenoxybenzyl)isonicotinohydrazide (N34red), were studied. Owing to their advantageous <em>in vitro</em> selectivity index against the primary mutation responsible for drug resistance in <em>Mycobacterium tuberculosis</em> (<em>Mtb</em>), as well as their suitable lipophilicity and interaction with human serum albumin, INH<img>C10 and N34 were deemed promising antitubercular compounds. N34red, despite differing from N34 only in the saturation of the <em>N</em>′ = C bond, presents a poor selectivity index. To delve deeper into the therapeutic potential of these compounds, their interaction with biomembrane models, mimicking biological barriers on the way to the target inside <em>Mtb</em> cells, was herein evaluated. All compounds, except N34red, weakened the packing of the acyl chains in the rigid lipid gel phase, especially INH<img>C10, which was the only compound disturbing liquid disordered membranes. Notably, all compounds except INH decreased membrane dipole potential, across all types of bilayers studied, but only N34red had a drastic effect. The insertion in gel phase bilayers suggests that the compounds may be able to penetrate the rigid cell wall of <em>Mtb</em>. Förster's resonance energy transfer (FRET) assays in ternary bilayers with liquid ordered/liquid disordered lateral heterogeneity mimicking human cell membranes, showed that the compounds affected neither the size nor the organization of lipid domains. These results provide molecular insights into the low toxicity against human cell lines and improved activity against drug-resistant <em>Mtb</em> of INH<img>C10 and N34.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"205 ","pages":"Article 106986"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142823788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.ejps.2024.106980
Nadina Zulbeari, Nanna Einshøj Lund, René Holm
The dual centrifugation approach has in the recent years emerged as a powerful milling tool to prepare pharmaceutical suspensions in submicron range with a fast-milling capacity by milling 40 samples simultaneously in 2 mL vials. While there is some standardized milling conditions described in the literature when preparing aqueous suspensions with dual centrifugation, a more systematic and experimental understanding of the milling process to evaluate the impact of different process variables in the dual centrifuge on the final sizes of the suspended drug particles independent of the drug compound used was desired. Overall, the present study demonstrated the applicability of the dual centrifuge for small-scale screening purposes and showed the impact of process parameters on the physical attributes of prepared suspensions. In the present work, the rate of size reduction on three different model compounds, i.e., cinnarizine, haloperidol, and indomethacin, was found to be mostly influenced by the milling speed, size of milling beads, and the bead loading during milling, whereas the rotor temperature did not affect the particle size profiles when stabilized with polysorbate 20 during milling with dual centrifugation. Smaller particle sizes were in general obtained at the highest milling intensity, i.e., 1500 rpm, smallest bead size, i.e., 0.2 mm, and higher bead loadings (42 %, 56 %, and 83 %). The grinding limit of approximately 0.50 µm, 0.70 µm, and 0.35 µm for cinnarizine, haloperidol, and indomethacin, respectively, was achieved relatively fast, i.e., 30 min of milling at the specified conditions, compared to when suspensions were milled with larger bead sizes (i.e., 1.0 mm), lower milling intensities (e.g., 1000 rpm), and lower bead loadings (e.g., 14 or 28 %). The study further confirmed that a higher milling intensity was necessary during milling of haloperidol suspensions probably due to the compounds predominantly plastic properties. Sizes of indomethacin particles increased with longer milling runs up to 240 min and also higher bead loadings of 56 % and 83 %. These observations were further supported by the color conversion from white to yellow of indomethacin suspensions which indicated generation of small quantities of amorphic material after milling with a high milling intensity. Upscale investigations showed comparable particle size profiles for all three model compounds while milling at 1500 rpm for five minutes.
{"title":"Small-scale aqueous suspension preparation using dual centrifugation: the effect of process parameters on the sizes of drug particles","authors":"Nadina Zulbeari, Nanna Einshøj Lund, René Holm","doi":"10.1016/j.ejps.2024.106980","DOIUrl":"10.1016/j.ejps.2024.106980","url":null,"abstract":"<div><div>The dual centrifugation approach has in the recent years emerged as a powerful milling tool to prepare pharmaceutical suspensions in submicron range with a fast-milling capacity by milling 40 samples simultaneously in 2 mL vials. While there is some standardized milling conditions described in the literature when preparing aqueous suspensions with dual centrifugation, a more systematic and experimental understanding of the milling process to evaluate the impact of different process variables in the dual centrifuge on the final sizes of the suspended drug particles independent of the drug compound used was desired. Overall, the present study demonstrated the applicability of the dual centrifuge for small-scale screening purposes and showed the impact of process parameters on the physical attributes of prepared suspensions. In the present work, the rate of size reduction on three different model compounds, <em>i.e.,</em> cinnarizine, haloperidol, and indomethacin, was found to be mostly influenced by the milling speed, size of milling beads, and the bead loading during milling, whereas the rotor temperature did not affect the particle size profiles when stabilized with polysorbate 20 during milling with dual centrifugation. Smaller particle sizes were in general obtained at the highest milling intensity, <em>i.e.,</em> 1500 rpm, smallest bead size, <em>i.e.,</em> 0.2 mm, and higher bead loadings (42 %, 56 %, and 83 %). The grinding limit of approximately 0.50 µm, 0.70 µm, and 0.35 µm for cinnarizine, haloperidol, and indomethacin, respectively, was achieved relatively fast, <em>i.e.,</em> 30 min of milling at the specified conditions, compared to when suspensions were milled with larger bead sizes (<em>i.e.,</em> 1.0 mm), lower milling intensities (<em>e.g.,</em> 1000 rpm), and lower bead loadings (<em>e.g.,</em> 14 or 28 %). The study further confirmed that a higher milling intensity was necessary during milling of haloperidol suspensions probably due to the compounds predominantly plastic properties. Sizes of indomethacin particles increased with longer milling runs up to 240 min and also higher bead loadings of 56 % and 83 %. These observations were further supported by the color conversion from white to yellow of indomethacin suspensions which indicated generation of small quantities of amorphic material after milling with a high milling intensity. Upscale investigations showed comparable particle size profiles for all three model compounds while milling at 1500 rpm for five minutes.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"205 ","pages":"Article 106980"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The formation of protein aggregates, which can be immunogenic and lower the efficacy and safety of protein drugs, has been an issue in biopharmaceutical development for more than a decade. Although protein drugs are often shipped as frozen material, the effect of the accidental dropping of frozen proteins, which can occur during shipping and handling, on the physical stability has not been studied. Here, a frozen Fc fusion protein was subjected to dropping stress and the increase in the aggregate concentration was evaluated. Significant increases in micron-sized aggregates were observed at –30 °C (p ≤ 0.01), but not at –60 °C. Proteins adsorbed on the vial surfaces were not remarkably detached by the action of dropping and were not the primary cause of the increase in micron-sized aggregates. When the vials were dropped, local heat generation occurred and this led to local freeze-thaw stress that induced protein aggregation. Poloxamer-188, which is known to mitigate aggregation caused by freeze-thaw stress, effectively prevented the aggregation caused by the dropping stress in the frozen state at –30 °C. In addition, rapid freezing could suppress the aggregation caused by the dropping stress. The results demonstrated that dropping stress reduced the stability of proteins even in the frozen state, and they provide new insights into the formulation and freezing processes to prevent protein aggregation caused by dropping stress in the frozen state.
{"title":"Protein aggregation in the frozen state induced by dropping stress","authors":"Tetsuo Torisu , Ayuko Maeda , Shuhei Ito , Susumu Uchiyama","doi":"10.1016/j.ejps.2024.106996","DOIUrl":"10.1016/j.ejps.2024.106996","url":null,"abstract":"<div><div>The formation of protein aggregates, which can be immunogenic and lower the efficacy and safety of protein drugs, has been an issue in biopharmaceutical development for more than a decade. Although protein drugs are often shipped as frozen material, the effect of the accidental dropping of frozen proteins, which can occur during shipping and handling, on the physical stability has not been studied. Here, a frozen Fc fusion protein was subjected to dropping stress and the increase in the aggregate concentration was evaluated. Significant increases in micron-sized aggregates were observed at –30 °C (<em>p</em> ≤ 0.01), but not at –60 °C. Proteins adsorbed on the vial surfaces were not remarkably detached by the action of dropping and were not the primary cause of the increase in micron-sized aggregates. When the vials were dropped, local heat generation occurred and this led to local freeze-thaw stress that induced protein aggregation. Poloxamer-188, which is known to mitigate aggregation caused by freeze-thaw stress, effectively prevented the aggregation caused by the dropping stress in the frozen state at –30 °C. In addition, rapid freezing could suppress the aggregation caused by the dropping stress. The results demonstrated that dropping stress reduced the stability of proteins even in the frozen state, and they provide new insights into the formulation and freezing processes to prevent protein aggregation caused by dropping stress in the frozen state.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"205 ","pages":"Article 106996"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142893253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-01DOI: 10.1016/j.ejps.2024.106992
Filip Gorachinov , Monika Koviloska , Katerina Tnokovska , Ana Atanasova , Packa Antovska , Jelena Lazova , Nikola Geskovski
This study leverages Fourier Transform Near-Infrared (FT-NIR) spectroscopy to monitor the coating process of pharmaceutical tablets using PVA-based TiO2-free films, with talc and iron oxides as opacifiers. By employing a combination of multivariate analytical techniques, the correlation between film coating progression and film thickness was evaluated. Assessment of coating thickness for different coating levels was performed by optical microscopy. Additionally, using colorimetric analysis by scanner method, the color progression for different coating levels was evaluated and expressed as the a* value from CIELAB color space. The coordinate value a* showed predictable changes with the progression of the coating process and film thickness values, indicating its utility as a robust reference method for quality control and process optimization. The predictive capability of the OPLS models, validated against measured film thickness and the a* value, demonstrated low prediction errors and confirmed the models' effectiveness in distinguishing coating levels and accurately predicting film coating progression. The OPLS model used knowledge-based peaks of interest, which were further confirmed by loading and coefficient plots. The study demonstrated that film thickness, as a destructive, and a* value from CIELAB color space, as a non-destructive reference method for coating progression could be used during a controlled pharmaceutical coating process for product quality assessment and pharmaceutical process endpoint determination.
{"title":"FT-NIR models for predicting film quality parameters in titanium dioxide-free tablet coatings","authors":"Filip Gorachinov , Monika Koviloska , Katerina Tnokovska , Ana Atanasova , Packa Antovska , Jelena Lazova , Nikola Geskovski","doi":"10.1016/j.ejps.2024.106992","DOIUrl":"10.1016/j.ejps.2024.106992","url":null,"abstract":"<div><div>This study leverages Fourier Transform Near-Infrared (FT-NIR) spectroscopy to monitor the coating process of pharmaceutical tablets using PVA-based TiO<sub>2</sub>-free films, with talc and iron oxides as opacifiers. By employing a combination of multivariate analytical techniques, the correlation between film coating progression and film thickness was evaluated. Assessment of coating thickness for different coating levels was performed by optical microscopy. Additionally, using colorimetric analysis by scanner method, the color progression for different coating levels was evaluated and expressed as the a* value from CIELAB color space. The coordinate value a* showed predictable changes with the progression of the coating process and film thickness values, indicating its utility as a robust reference method for quality control and process optimization. The predictive capability of the OPLS models, validated against measured film thickness and the a* value, demonstrated low prediction errors and confirmed the models' effectiveness in distinguishing coating levels and accurately predicting film coating progression. The OPLS model used knowledge-based peaks of interest, which were further confirmed by loading and coefficient plots. The study demonstrated that film thickness, as a destructive, and a* value from CIELAB color space, as a non-destructive reference method for coating progression could be used during a controlled pharmaceutical coating process for product quality assessment and pharmaceutical process endpoint determination.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"205 ","pages":"Article 106992"},"PeriodicalIF":4.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142853390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-27DOI: 10.1016/j.ejps.2025.107026
Armin Hauk , Alexander Wildschütz , Ina Pahl , Daniel Canton , Roberto Menzel
We evaluated algorithms designed to extrapolate extractables data for predicting process equipment-related leachables (PERLs) and assessing PERL exposure in single-use systems (SUSs) and assemblies. The robustness and sensitivity of these algorithms were tested against variations in input data, including extrapolation algorithms for both short and long contact time extractables data obtained from the standardized extractables protocol provided in USP 〈665〉. Our findings demonstrate that extrapolated data for SUS and assemblies are suitable for safety assessments. Extrapolated and aggregated data do not systematically underestimate potential PERL exposure values, provided that the extractables data originate from experiments with a higher surface area to contact liquid volume ratio and/or a low liquid to material volume ratio compared to the use scenario. The algorithms are non-sensitive to deviations in input data, as these deviations are propagated decreasingly into extrapolated data and parameters. The quality and significance of PERL exposure calculations can be enhanced by incorporating extractables study data from experiments using a semipolar organic solution, such as ethanol.
{"title":"From extractables to exposure data: Sensitivity analysis of extrapolation algorithms with focus on USP 〈665〉","authors":"Armin Hauk , Alexander Wildschütz , Ina Pahl , Daniel Canton , Roberto Menzel","doi":"10.1016/j.ejps.2025.107026","DOIUrl":"10.1016/j.ejps.2025.107026","url":null,"abstract":"<div><div>We evaluated algorithms designed to extrapolate extractables data for predicting process equipment-related leachables (PERLs) and assessing PERL exposure in single-use systems (SUSs) and assemblies. The robustness and sensitivity of these algorithms were tested against variations in input data, including extrapolation algorithms for both short and long contact time extractables data obtained from the standardized extractables protocol provided in USP 〈665〉. Our findings demonstrate that extrapolated data for SUS and assemblies are suitable for safety assessments. Extrapolated and aggregated data do not systematically underestimate potential PERL exposure values, provided that the extractables data originate from experiments with a higher surface area to contact liquid volume ratio and/or a low liquid to material volume ratio compared to the use scenario. The algorithms are non-sensitive to deviations in input data, as these deviations are propagated decreasingly into extrapolated data and parameters. The quality and significance of PERL exposure calculations can be enhanced by incorporating extractables study data from experiments using a semipolar organic solution, such as ethanol.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"207 ","pages":"Article 107026"},"PeriodicalIF":4.3,"publicationDate":"2025-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143046026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-24DOI: 10.1016/j.ejps.2025.107025
Pedro G M Canhão, Jan Snoeys, Suzy Geerinckx, Marjolein van Heerden, An Van den Bergh, Camden Holm, Jan Markus, Seyoum Ayehunie, Mario Monshouwer, Raymond Evers, Patrick Augustijns, Stephanie Kourula
The purpose of this study was to evaluate EpiColon, a novel human organotypic 3D colon microtissue prototype, developed to assess colonic drug disposition, with a particular focus on permeability ranking, and compare its performance to Caco-2 monolayers. EpiColon was characterized for barrier function using transepithelial electrical resistance (TEER), morphology via histology and immunohistochemistry, and functionality through drug transport studies measuring apparent permeability (Papp). Cutoff thresholds for the permeability of FITC-dextran 4 kDa (FD4), FITC-dextran 10 kDa (FD10S), and [14C]mannitol were established to monitor microtissue integrity. Permeability of EpiColon for 20 benchmark drugs was compared with Caco-2 data, and the activity of pivotal efflux transporters, including multidrug resistance protein 1/P-glycoprotein (MDR1/P-gp), along with multidrug resistance protein 2 (MRP2) and breast cancer resistance protein (BCRP), was evaluated using selective substrates. EpiColon exhibited a physiological barrier function (272.0 ± 53.05 Ω x cm2) and effectively discriminated between high (e.g., budesonide and [3H]metoprolol) and low permeable compounds (e.g., [3H]atenolol and [14C]mannitol). The model demonstrated functional activity for key efflux transporters, with efflux ratios of 2.32 for [3H]digoxin (MDR1/P-gp) and 3.34 for sulfasalazine (MRP2 and BCRP). Notably, EpiColon showed an enhanced dynamic range in the low permeability range, differentiating Papp between FD4 and FD10S, in contrast to Caco-2 monolayers. Significant positive correlations were observed between human fraction absorbed (fabs) and logarithmically transformed Papp [AP-BL] values for both EpiColon (rs = 0.68) and Caco-2 (rs = 0.68). Furthermore, EpiColon recapitulates some essential phenotypic and cellular features of the human colon, including the expression of critical marker genes (Pan-Cytokeratin+: epithelial/colonocytes, Vimentin+: mesenchymal/fibroblast, and Alcian Blue+: goblet cell/mucus). In conclusion, EpiColon is a promising platform that offers a valuable complement to conventional Caco-2 monolayers for studying colonic drug disposition. However, the presence of flat and some cuboidal cells, along with low throughput, must be addressed to improve its applicability in both academic research and pharmaceutical industry.
{"title":"Human organotypic colon in vitro microtissue: unveiling a new window into colonic drug disposition.","authors":"Pedro G M Canhão, Jan Snoeys, Suzy Geerinckx, Marjolein van Heerden, An Van den Bergh, Camden Holm, Jan Markus, Seyoum Ayehunie, Mario Monshouwer, Raymond Evers, Patrick Augustijns, Stephanie Kourula","doi":"10.1016/j.ejps.2025.107025","DOIUrl":"https://doi.org/10.1016/j.ejps.2025.107025","url":null,"abstract":"<p><p>The purpose of this study was to evaluate EpiColon, a novel human organotypic 3D colon microtissue prototype, developed to assess colonic drug disposition, with a particular focus on permeability ranking, and compare its performance to Caco-2 monolayers. EpiColon was characterized for barrier function using transepithelial electrical resistance (TEER), morphology via histology and immunohistochemistry, and functionality through drug transport studies measuring apparent permeability (P<sub>app</sub>). Cutoff thresholds for the permeability of FITC-dextran 4 kDa (FD4), FITC-dextran 10 kDa (FD10S), and [<sup>14</sup>C]mannitol were established to monitor microtissue integrity. Permeability of EpiColon for 20 benchmark drugs was compared with Caco-2 data, and the activity of pivotal efflux transporters, including multidrug resistance protein 1/P-glycoprotein (MDR1/P-gp), along with multidrug resistance protein 2 (MRP2) and breast cancer resistance protein (BCRP), was evaluated using selective substrates. EpiColon exhibited a physiological barrier function (272.0 ± 53.05 Ω x cm<sup>2</sup>) and effectively discriminated between high (e.g., budesonide and [<sup>3</sup>H]metoprolol) and low permeable compounds (e.g., [<sup>3</sup>H]atenolol and [<sup>14</sup>C]mannitol). The model demonstrated functional activity for key efflux transporters, with efflux ratios of 2.32 for [<sup>3</sup>H]digoxin (MDR1/P-gp) and 3.34 for sulfasalazine (MRP2 and BCRP). Notably, EpiColon showed an enhanced dynamic range in the low permeability range, differentiating P<sub>app</sub> between FD4 and FD10S, in contrast to Caco-2 monolayers. Significant positive correlations were observed between human fraction absorbed (f<sub>abs</sub>) and logarithmically transformed P<sub>app</sub> [AP-BL] values for both EpiColon (r<sub>s</sub> = 0.68) and Caco-2 (r<sub>s</sub> = 0.68). Furthermore, EpiColon recapitulates some essential phenotypic and cellular features of the human colon, including the expression of critical marker genes (Pan-Cytokeratin<sup>+</sup>: epithelial/colonocytes, Vimentin<sup>+</sup>: mesenchymal/fibroblast, and Alcian Blue<sup>+</sup>: goblet cell/mucus). In conclusion, EpiColon is a promising platform that offers a valuable complement to conventional Caco-2 monolayers for studying colonic drug disposition. However, the presence of flat and some cuboidal cells, along with low throughput, must be addressed to improve its applicability in both academic research and pharmaceutical industry.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"107025"},"PeriodicalIF":4.3,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143046028","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}
Premature drug release is the primary hindrance to the effective function of the lyso-thermosensitive liposomes (LTSLs) of doxorubicin (Dox), known as ThermoDox® for the treatment of cancer. Herein, we have optimized LTSLs by using a combination of phospholipids (PLs) with high transition temperatures (Tm) to improve the therapeutic outcome in an assisted ultrasound approach. For this, several Dox LTSLs were prepared using the remote loading method at varying molar ratios (0 to 90 %) of DPPC (Tm 41 °C) and HSPC (Tm 54.5 °C), as well as a constant molar ratio of MSPC (10 %), DSPE-mPEG2000 (4 %). The treatment efficacy was explored by using ultrasound as external hyperthermia (HT) (40–42℃) in mice bearing C26 murine colon carcinoma. All the formulations had an average diameter of around 110 nm, PDI ≤ 0.15, zeta potential of around -12 mV, and Dox encapsulation of >90 %. The cytotoxicity results indicated a higher IC50 value of Dox-LTSLs compared to the ThermoDox® (F0: DPPC:MSPC:DSPE-mPEG2000, 90:10:4), attributed to the faster Dox release in F0 formulation devoid of HSPC. Among various formulations, F25 (DPPC: MSPC: DSPE-mPEG2000: HSPC, 65:10:4:25) showed the highest cellular uptake at 42℃ and significantly improved the antitumor and survival efficacy in mice bearing C26 colon carcinoma in combination with ultrasonic HT compared to F0. Collectively, results demonstrated that optimizing the rigidity of the liposomal bilayers through the combinatorial selection of PLs of different transition temperatures could improve the plasma stability of the liposome, and hence ameliorate the outcome of therapy in assistance with an effective HT approach.
{"title":"Ultrasound-assisted efficient targeting of doxorubicin to the tumor microenvironment by lyso-thermosensitive liposomes of varying phase transition temperatures","authors":"Seyedeh Maryam Hosseinikhah , Leila Farhoudi , Farshad Mirzavi , Fatemeh Vahdat-Lasemi , Leila Arabi , Fatemeh Gheybi , Ameneh Sazgarnia , Seyedeh Hoda Alavizadeh , Mahmoud Reza Jaafari","doi":"10.1016/j.ejps.2025.107024","DOIUrl":"10.1016/j.ejps.2025.107024","url":null,"abstract":"<div><div>Premature drug release is the primary hindrance to the effective function of the lyso-thermosensitive liposomes (LTSLs) of doxorubicin (Dox), known as ThermoDox® for the treatment of cancer. Herein, we have optimized LTSLs by using a combination of phospholipids (PLs) with high transition temperatures (Tm) to improve the therapeutic outcome in an assisted ultrasound approach. For this, several Dox LTSLs were prepared using the remote loading method at varying molar ratios (0 to 90 %) of DPPC (Tm 41 °C) and HSPC (Tm 54.5 °C), as well as a constant molar ratio of MSPC (10 %), DSPE-mPEG<sub>2000</sub> (4 %). The treatment efficacy was explored by using ultrasound as external hyperthermia (HT) (40–42℃) in mice bearing C26 murine colon carcinoma. All the formulations had an average diameter of around 110 nm, PDI ≤ 0.15, zeta potential of around -12 mV, and Dox encapsulation of >90 %. The cytotoxicity results indicated a higher IC<sub>50</sub> value of Dox-LTSLs compared to the ThermoDox® (F0: DPPC:MSPC:DSPE-mPEG<sub>2000</sub>, 90:10:4), attributed to the faster Dox release in F0 formulation devoid of HSPC. Among various formulations, F25 (DPPC: MSPC: DSPE-mPEG<sub>2000</sub>: HSPC, 65:10:4:25) showed the highest cellular uptake at 42℃ and significantly improved the antitumor and survival efficacy in mice bearing C26 colon carcinoma in combination with ultrasonic HT compared to F0. Collectively, results demonstrated that optimizing the rigidity of the liposomal bilayers through the combinatorial selection of PLs of different transition temperatures could improve the plasma stability of the liposome, and hence ameliorate the outcome of therapy in assistance with an effective HT approach.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"206 ","pages":"Article 107024"},"PeriodicalIF":4.3,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143037653","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-21DOI: 10.1016/j.ejps.2025.107017
Morenikeji Aina , Darya Kuznyetsova , Fabien Baillon , Romain Sescousse , Noelia M. Sanchez-Ballester , Sylvie Begu , Ian Soulairol , Martial Sauceau
This study investigates the effects of disintegrants sodium starch glycolate (SSG) and crospovidone (CP) on the printability, rheological properties, and disintegration time of agar and hydroxypropyl methylcellulose (HPMC)-based formulations designed for semi-solid extrusion. Printability was assessed by measuring the dimensional accuracy of manually extruded filaments. Rheological analysis was performed using oscillatory measurements. Principal component analysis (PCA) and Spearman correlation analysis identified three key components (phase angle, critical strain, and elastic modulus) that explained the total variance in the rheological dataset. A 2 × 3 factorial design was employed to evaluate the impact of CP, SSG, and HPMC on these rheological parameters, as well as on printability and disintegration time. Results indicated that formulations containing HPMC and SSG generally exhibited better printability. Formulations containing CP achieved satisfactory printability only when SSG or HPMC was included. The optimal printability and rheological properties were achieved with formulations containing 5 % CP and 10 % SSG. Linear regression models correlated geometric volumes of the model and pycnometric volumes of printed objects, with validation showing that predicted masses were within a 95 % confidence interval of measured values for various shapes. All formulations demonstrated immediate-release properties, confirming the successful fabrication of personalised immediate-release dosage forms using semi-solid extrusion technology.
{"title":"Impact of disintegrants on rheological properties and printability in SSE 3D printing of immediate-release formulations","authors":"Morenikeji Aina , Darya Kuznyetsova , Fabien Baillon , Romain Sescousse , Noelia M. Sanchez-Ballester , Sylvie Begu , Ian Soulairol , Martial Sauceau","doi":"10.1016/j.ejps.2025.107017","DOIUrl":"10.1016/j.ejps.2025.107017","url":null,"abstract":"<div><div>This study investigates the effects of disintegrants sodium starch glycolate (SSG) and crospovidone (CP) on the printability, rheological properties, and disintegration time of agar and hydroxypropyl methylcellulose (HPMC)-based formulations designed for semi-solid extrusion. Printability was assessed by measuring the dimensional accuracy of manually extruded filaments. Rheological analysis was performed using oscillatory measurements. Principal component analysis (PCA) and Spearman correlation analysis identified three key components (phase angle, critical strain, and elastic modulus) that explained the total variance in the rheological dataset. A 2 × 3<span><math><msup><mrow></mrow><mrow><mn>2</mn></mrow></msup></math></span> factorial design was employed to evaluate the impact of CP, SSG, and HPMC on these rheological parameters, as well as on printability and disintegration time. Results indicated that formulations containing HPMC and SSG generally exhibited better printability. Formulations containing CP achieved satisfactory printability only when SSG or HPMC was included. The optimal printability and rheological properties were achieved with formulations containing 5 % CP and 10 % SSG. Linear regression models correlated geometric volumes of the model and pycnometric volumes of printed objects, with validation showing that predicted masses were within a 95 % confidence interval of measured values for various shapes. All formulations demonstrated immediate-release properties, confirming the successful fabrication of personalised immediate-release dosage forms using semi-solid extrusion technology.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"206 ","pages":"Article 107017"},"PeriodicalIF":4.3,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-21DOI: 10.1016/j.ejps.2025.107023
Rui Silva , Helena Colom , Anabela Almeida , Joana Bicker , Andreia Carona , Ana Silva , Francisco Sales , Isabel Santana , Amílcar Falcão , Ana Fortuna
Zonisamide exhibits significant pharmacokinetic variability, demanding for the development of population pharmacokinetic (PopPK) models to identify key factors influencing drug disposition.
This study aimed to develop and validate a PopPK model to optimize zonisamide posology in patients with refractory epilepsy.
A total of 114 plasma concentrations of zonisamide, obtained from 64 patients, were used for PopPK model development, employing the nonlinear mixed-effects modelling approach. The final model was evaluated by visually inspecting the goodness-of-fit plots and the visual predictive check plot and by the bootstrap resampling method. A one-compartment model with first-order elimination was the one that best described the pharmacokinetic profile of zonisamide. Between-patient variability (BPV) was included on clearance (CL/F), volume of distribution (Vd/F) and absorption rate constant (ka). The residual error (RE) was modeled as proportional. The final model estimates for CL/F, Vd/F and ka were 0.761 L/h, 48.10 L and 0.671 h⁻¹, respectively. The BPV associated with CL/F, Vd/F, and ka was 43.93%, 52.06%, and 91.27%, respectively, while the proportional RE was 7.18%. The concomitant administration of enzyme-inducing antiseizure drugs (EIASDs), included in the model as inducer drug load (INDDL), significantly accounted for BPV associated with CL/F and led to increased CL/F in patients receiving EIASDs compared to the others. Consequently, patients receiving EIASDs require higher daily doses of zonisamide to achieve therapeutic plasma concentrations compared to those not treated with EIASDs.
Model validation, using bootstrap and visual predictive checks, confirmed its stability and robustness, making it a valuable tool for individualized zonisamide dosing in adults with refractory epilepsy.
{"title":"A new population pharmacokinetic model for dosing optimization of zonisamide in patients with refractory epilepsy","authors":"Rui Silva , Helena Colom , Anabela Almeida , Joana Bicker , Andreia Carona , Ana Silva , Francisco Sales , Isabel Santana , Amílcar Falcão , Ana Fortuna","doi":"10.1016/j.ejps.2025.107023","DOIUrl":"10.1016/j.ejps.2025.107023","url":null,"abstract":"<div><div>Zonisamide exhibits significant pharmacokinetic variability, demanding for the development of population pharmacokinetic (PopPK) models to identify key factors influencing drug disposition.</div><div>This study aimed to develop and validate a PopPK model to optimize zonisamide posology in patients with refractory epilepsy.</div><div>A total of 114 plasma concentrations of zonisamide, obtained from 64 patients, were used for PopPK model development, employing the nonlinear mixed-effects modelling approach. The final model was evaluated by visually inspecting the goodness-of-fit plots and the visual predictive check plot and by the bootstrap resampling method. A one-compartment model with first-order elimination was the one that best described the pharmacokinetic profile of zonisamide. Between-patient variability (BPV) was included on clearance (CL/F), volume of distribution (Vd/F) and absorption rate constant (ka). The residual error (RE) was modeled as proportional. The final model estimates for CL/F, Vd/F and ka were 0.761 L/h, 48.10 L and 0.671 h⁻¹, respectively. The BPV associated with CL/F, Vd/F, and ka was 43.93%, 52.06%, and 91.27%, respectively, while the proportional RE was 7.18%. The concomitant administration of enzyme-inducing antiseizure drugs (EIASDs), included in the model as inducer drug load (INDDL), significantly accounted for BPV associated with CL/F and led to increased CL/F in patients receiving EIASDs compared to the others. Consequently, patients receiving EIASDs require higher daily doses of zonisamide to achieve therapeutic plasma concentrations compared to those not treated with EIASDs.</div><div>Model validation, using bootstrap and visual predictive checks, confirmed its stability and robustness, making it a valuable tool for individualized zonisamide dosing in adults with refractory epilepsy.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"207 ","pages":"Article 107023"},"PeriodicalIF":4.3,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-17DOI: 10.1016/j.ejps.2025.107018
Sandi Svetič , Laura Medved , Franc Vrečer , Klemen Korasa
One of the main concerns with formulations containing amorphous solid dispersions (ASDs) is their physical stability. Stability can be compromised if a formulation contains any residual crystallinity of an active pharmaceutical ingredient (API) that could act as seeds for further crystallisation. This study presents four methods for crystalline amlodipine maleate quantification in ASD, which were developed using one Raman and three NIR process analysers. A preliminary analysis revealed distinct differences between amorphous and non-amorphous forms of the API, both in the API alone and in the formulation. These differences laid the foundation for model development in subsequent steps. The development of four partial least squares (PLS) models proceeded through two stages, initially using a single granulation batch dataset for training, and then expanding to include three batches. Their predictability was evaluated on an additional batch dataset. Models were evaluated primarily using root mean square error of prediction (RMSEP), residual prediction deviation (RPD), and limit of detection along with other metrics. To the best of authors’ knowledge, this is the first study that focuses on process monitoring of fluidized bed granulation used in preparation of ASD. The results of this study and their interpretations present novel aspects of Raman and NIR process analyser applications in combination with PLS.
{"title":"Quantifying crystallinity of amlodipine maleate in amorphous solid dispersions produced by fluidized bed granulation using PAT tools","authors":"Sandi Svetič , Laura Medved , Franc Vrečer , Klemen Korasa","doi":"10.1016/j.ejps.2025.107018","DOIUrl":"10.1016/j.ejps.2025.107018","url":null,"abstract":"<div><div>One of the main concerns with formulations containing amorphous solid dispersions (ASDs) is their physical stability. Stability can be compromised if a formulation contains any residual crystallinity of an active pharmaceutical ingredient (API) that could act as seeds for further crystallisation. This study presents four methods for crystalline amlodipine maleate quantification in ASD, which were developed using one Raman and three NIR process analysers. A preliminary analysis revealed distinct differences between amorphous and non-amorphous forms of the API, both in the API alone and in the formulation. These differences laid the foundation for model development in subsequent steps. The development of four partial least squares (PLS) models proceeded through two stages, initially using a single granulation batch dataset for training, and then expanding to include three batches. Their predictability was evaluated on an additional batch dataset. Models were evaluated primarily using root mean square error of prediction (RMSEP), residual prediction deviation (RPD), and limit of detection along with other metrics. To the best of authors’ knowledge, this is the first study that focuses on process monitoring of fluidized bed granulation used in preparation of ASD. The results of this study and their interpretations present novel aspects of Raman and NIR process analyser applications in combination with PLS.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"206 ","pages":"Article 107018"},"PeriodicalIF":4.3,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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