Eva J. Streekstra , Marit Keuper-Navis , Jeroen J.W.M. van den Heuvel , Petra van den Broek , Rick Greupink , Martijn W.J. Stommel , Willem P. de Boode , Sanne M.B.I. Botden , Frans G.M. Russel , Evita van de Steeg , Saskia N. de Wildt
{"title":"从儿童和成人中提取的肠溶物在研究肠道 CYP3A4/5 代谢的年龄依赖性差异方面的潜力。","authors":"Eva J. Streekstra , Marit Keuper-Navis , Jeroen J.W.M. van den Heuvel , Petra van den Broek , Rick Greupink , Martijn W.J. Stommel , Willem P. de Boode , Sanne M.B.I. Botden , Frans G.M. Russel , Evita van de Steeg , Saskia N. de Wildt","doi":"10.1016/j.ejps.2024.106868","DOIUrl":null,"url":null,"abstract":"<div><p>Drug metabolism in the intestinal wall affects bioavailability of orally administered drugs and is influenced by age. Hence, it is important to fully understand the drug metabolizing capacity of the gut to predict systemic exposure. The aim of this study was to investigate the potential of enteroids as a tool to study CYP3A4/5 -mediated metabolism in both children and adults.</p><p>Bioconversion of midazolam, a CYP3A4/5 model substrate, was studied using enteroid monolayers as well as tissue explants in the Ussing chamber, both derived from pediatric [median (range age): 54 weeks (2 days – 13 years), <em>n</em> = 21] and adult (<em>n</em> = 5) tissue. Caco-2 cellular monolayers were employed as controls. In addition, mRNA expression of CYP3A4 was determined in enteroid monolayers (<em>n</em> = 11), tissue (<em>n</em> = 23) and Caco-2 using RT-qPCR.</p><p>Midazolam metabolism was successfully detected in all enteroid monolayers, as well as in all tissue explants studied in the Ussing chamber, whereas Caco-2 showed no significant metabolite formation. The extracted fraction of midazolam was similar between enteroid monolayers and tissue. The fraction of midazolam extracted increased with age in enteroid monolayers derived from 0 to 70 week old donors. No statistically significant correlation was observed in tissue likely due to high variability observed and the smaller donor numbers included in the study. At the level of gene expression, CYP3A4 increased with age in tissues (<em>n</em> = 32), while this was not reflected in enteroid monolayers (<em>n</em> = 16). Notably, asymmetric metabolite formation was observed in enteroids and tissue, with higher metabolite formation on the luminal side of the barrier.</p><p>In summary, we demonstrated that enteroids can be used to measure CYP3A4/5 midazolam metabolism, which we show is similar as observed in fresh isolated tissue. This was the case both in children and adults, indicating the potential of enteroids to predict intestinal metabolism. This study provides promising data to further develop enteroids to study drug metabolism <em>in vitro</em> and potentially predict oral absorption for special populations as an alternative to using fresh tissue.</p></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"201 ","pages":"Article 106868"},"PeriodicalIF":4.3000,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0928098724001805/pdfft?md5=1296665988e874307519ccd3ae0bcd79&pid=1-s2.0-S0928098724001805-main.pdf","citationCount":"0","resultStr":"{\"title\":\"The potential of enteroids derived from children and adults to study age-dependent differences in intestinal CYP3A4/5 metabolism\",\"authors\":\"Eva J. Streekstra , Marit Keuper-Navis , Jeroen J.W.M. van den Heuvel , Petra van den Broek , Rick Greupink , Martijn W.J. 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In addition, mRNA expression of CYP3A4 was determined in enteroid monolayers (<em>n</em> = 11), tissue (<em>n</em> = 23) and Caco-2 using RT-qPCR.</p><p>Midazolam metabolism was successfully detected in all enteroid monolayers, as well as in all tissue explants studied in the Ussing chamber, whereas Caco-2 showed no significant metabolite formation. The extracted fraction of midazolam was similar between enteroid monolayers and tissue. The fraction of midazolam extracted increased with age in enteroid monolayers derived from 0 to 70 week old donors. No statistically significant correlation was observed in tissue likely due to high variability observed and the smaller donor numbers included in the study. At the level of gene expression, CYP3A4 increased with age in tissues (<em>n</em> = 32), while this was not reflected in enteroid monolayers (<em>n</em> = 16). 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The potential of enteroids derived from children and adults to study age-dependent differences in intestinal CYP3A4/5 metabolism
Drug metabolism in the intestinal wall affects bioavailability of orally administered drugs and is influenced by age. Hence, it is important to fully understand the drug metabolizing capacity of the gut to predict systemic exposure. The aim of this study was to investigate the potential of enteroids as a tool to study CYP3A4/5 -mediated metabolism in both children and adults.
Bioconversion of midazolam, a CYP3A4/5 model substrate, was studied using enteroid monolayers as well as tissue explants in the Ussing chamber, both derived from pediatric [median (range age): 54 weeks (2 days – 13 years), n = 21] and adult (n = 5) tissue. Caco-2 cellular monolayers were employed as controls. In addition, mRNA expression of CYP3A4 was determined in enteroid monolayers (n = 11), tissue (n = 23) and Caco-2 using RT-qPCR.
Midazolam metabolism was successfully detected in all enteroid monolayers, as well as in all tissue explants studied in the Ussing chamber, whereas Caco-2 showed no significant metabolite formation. The extracted fraction of midazolam was similar between enteroid monolayers and tissue. The fraction of midazolam extracted increased with age in enteroid monolayers derived from 0 to 70 week old donors. No statistically significant correlation was observed in tissue likely due to high variability observed and the smaller donor numbers included in the study. At the level of gene expression, CYP3A4 increased with age in tissues (n = 32), while this was not reflected in enteroid monolayers (n = 16). Notably, asymmetric metabolite formation was observed in enteroids and tissue, with higher metabolite formation on the luminal side of the barrier.
In summary, we demonstrated that enteroids can be used to measure CYP3A4/5 midazolam metabolism, which we show is similar as observed in fresh isolated tissue. This was the case both in children and adults, indicating the potential of enteroids to predict intestinal metabolism. This study provides promising data to further develop enteroids to study drug metabolism in vitro and potentially predict oral absorption for special populations as an alternative to using fresh tissue.
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The journal publishes research articles, review articles and scientific commentaries on all aspects of the pharmaceutical sciences with emphasis on conceptual novelty and scientific quality. The Editors welcome articles in this multidisciplinary field, with a focus on topics relevant for drug discovery and development.
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