{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T16:51:58Z","timestamp":1788367918963,"version":"build-2803163510"},"reference-count":70,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2022,2,17]],"date-time":"2022-02-17T00:00:00Z","timestamp":1645056000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010767","name":"Innovative Medicines Initiative","doi-asserted-by":"publisher","award":["116030"],"award-info":[{"award-number":["116030"]}],"id":[{"id":"10.13039\/501100010767","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Gefitinib is a tyrosine kinase inhibitor (TKI) that selectively inhibits the epidermal growth factor receptor (EGFR), hampering cell growth and proliferation. Due to its action, gefitinib has been used in the treatment of cancers that present abnormally increased expression of EGFR. However, side effects from gefitinib therapy may occur, among which diarrhoea is most common, that can lead to interruption of the planned therapy in the more severe cases. The mechanisms underlying intestinal toxicity induced by gefitinib are not well understood. Therefore, this study aims at providing insight into these mechanisms based on transcriptomic responses induced in vitro. A 3D culture of healthy human colon and small intestine (SI) organoids was exposed to 0.1, 1, 10 and 30 \u00b5M of gefitinib, for a maximum of three days. These drug concentrations were selected using physiologically-based pharmacokinetic simulation considering patient dosing regimens. Samples were used for the analysis of viability and caspase 3\/7 activation, image-based analysis of structural changes, as well as RNA isolation and sequencing via high-throughput techniques. Differential gene expression analysis showed that gefitinib perturbed signal transduction pathways, apoptosis, cell cycle, FOXO-mediated transcription, p53 signalling pathway, and metabolic pathways. Remarkably, opposite expression patterns of genes associated with metabolism of lipids and cholesterol biosynthesis were observed in colon versus SI organoids in response to gefitinib. These differences in the organoids\u2019 responses could be linked to increased activated protein kinase (AMPK) activity in colon, which can influence the sensitivity of the colon to the drug. Therefore, this study sheds light on how gefitinib induces toxicity in intestinal organoids and provides an avenue towards the development of a potential tool for drug screening and development.<\/jats:p>","DOI":"10.3390\/ijms23042213","type":"journal-article","created":{"date-parts":[[2022,2,17]],"date-time":"2022-02-17T20:25:45Z","timestamp":1645129545000},"page":"2213","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["A Transcriptomic Approach to Elucidate the Mechanisms of Gefitinib-Induced Toxicity in Healthy Human Intestinal Organoids"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5527-6627","authenticated-orcid":false,"given":"Daniela","family":"Rodrigues","sequence":"first","affiliation":[{"name":"Department of Toxicogenomics, GROW School for Oncology and Developmental Biology, Maastricht University, 6229 ER Maastricht, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bram","family":"Herpers","sequence":"additional","affiliation":[{"name":"Crown Bioscience Netherlands B.V., J.H. Oortweg 21, 2333 CH Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofia","family":"Ferreira","sequence":"additional","affiliation":[{"name":"Simcyp Division, Certara UK Limited, Sheffield S1 2BJ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heeseung","family":"Jo","sequence":"additional","affiliation":[{"name":"Simcyp Division, Certara UK Limited, Sheffield S1 2BJ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ciar\u00e1n","family":"Fisher","sequence":"additional","affiliation":[{"name":"Simcyp Division, Certara UK Limited, Sheffield S1 2BJ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luke","family":"Coyle","sequence":"additional","affiliation":[{"name":"Boehringer Ingelheim International GmbH, Pharmaceuticals Inc., Ridgefield, CT 06877, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seung-Wook","family":"Chung","sequence":"additional","affiliation":[{"name":"Boehringer Ingelheim International GmbH, Pharmaceuticals Inc., Ridgefield, CT 06877, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos C. S.","family":"Kleinjans","sequence":"additional","affiliation":[{"name":"Department of Toxicogenomics, GROW School for Oncology and Developmental Biology, Maastricht University, 6229 ER Maastricht, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8618-2487","authenticated-orcid":false,"given":"Danyel G. 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Drug Metab."},{"key":"ref_4","first-page":"282","article-title":"Pharmacokinetics of Gefitinib: Roles of Drug Metabolizing Enzymes and Transporters","volume":"14","author":"Zhao","year":"2017","journal-title":"Curr. Drug Deliv."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1110","DOI":"10.1016\/j.jaad.2017.07.048","article-title":"Gefitinib for patients with incurable cutaneous squamous cell carcinoma: A single-arm phase II clinical trial","volume":"77","author":"William","year":"2017","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.ygyno.2007.07.057","article-title":"A phase II trial to evaluate gefitinib as second- or third-line treatment in patients with recurring locoregionally advanced or metastatic cervical cancer","volume":"108","author":"Goncalves","year":"2008","journal-title":"Gynecol. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/s12032-016-0773-5","article-title":"Effects of polymorphisms in CYP2D6 and ABC transporters and side effects induced by gefitinib on the pharmacokinetics of the gefitinib metabolite, O-desmethyl gefitinib","volume":"33","author":"Kobayashi","year":"2016","journal-title":"Med. Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5170","DOI":"10.1158\/1078-0432.CCR-07-0574","article-title":"The intestinotrophic peptide, glp-2, counteracts intestinal atrophy in mice induced by the epidermal growth factor receptor inhibitor, gefitinib","volume":"13","author":"Hare","year":"2007","journal-title":"Clin. Cancer Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1634\/theoncologist.2014-0154","article-title":"Gefitinib and Erlotinib in Metastatic Non-Small Cell Lung Cancer: A Meta-Analysis of Toxicity and Efficacy of Randomized Clinical Trials","volume":"20","author":"Burotto","year":"2015","journal-title":"Oncologist"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3","DOI":"10.3892\/mco.2016.1099","article-title":"Toxicity profile of epidermal growth factor receptor tyrosine kinase inhibitors in patients with epidermal growth factor receptor gene mutation-positive lung cancer (Review)","volume":"6","author":"Takeda","year":"2017","journal-title":"Mol. Clin. Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"8531","DOI":"10.1200\/JCO.2004.00.6650","article-title":"Side effects related to cancer treatment: CASE 1. Hepatitis following treatment with gefitinib","volume":"23","author":"Ho","year":"2005","journal-title":"J. Clin. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2504","DOI":"10.1200\/JCO.2004.09.064","article-title":"Side effects of therapy: Case 1. Nephrotic syndrome associated with gefitinib therapy","volume":"22","author":"Kumasaka","year":"2004","journal-title":"J. Clin. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"366","DOI":"10.3816\/CLC.2003.n.016","article-title":"Dermatologic side effects associated with gefitinib therapy: Clinical experience and management","volume":"4","author":"Herbst","year":"2003","journal-title":"Clin. Lung Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2691","DOI":"10.1007\/s00204-021-03092-2","article-title":"New insights into the mechanisms underlying 5-fluorouracil-induced intestinal toxicity based on transcriptomic and metabolomic responses in human intestinal organoids","volume":"95","author":"Rodrigues","year":"2021","journal-title":"Arch. Toxicol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1065","DOI":"10.4252\/wjsc.v11.i12.1065","article-title":"Three-dimensional cell culture systems as an in vitro platform for cancer and stem cell modeling","volume":"11","author":"Chaicharoenaudomrung","year":"2019","journal-title":"World J. Stem Cells"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Nii, T., Makino, K., and Tabata, Y. (2020). Three-Dimensional Culture System of Cancer Cells Combined with Biomaterials for Drug Screening. Cancers, 12.","DOI":"10.3390\/cancers12102754"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1038\/nature07935","article-title":"Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche","volume":"459","author":"Sato","year":"2009","journal-title":"Nature"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3759","DOI":"10.1074\/jbc.R114.635995","article-title":"Human Enteroids\/Colonoids and Intestinal Organoids Functionally Recapitulate Normal Intestinal Physiology and Pathophysiology","volume":"291","author":"Zachos","year":"2016","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1038\/ncb3312","article-title":"Organoids as an in vitro model of human development and disease","volume":"18","author":"Fatehullah","year":"2016","journal-title":"Nat. Cell Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1038\/s41580-020-0259-3","article-title":"Human organoids: Model systems for human biology and medicine","volume":"21","author":"Kim","year":"2020","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1186\/s13567-021-00931-z","article-title":"Cerebral organoids and their potential for studies of brain diseases in domestic animals","volume":"52","author":"Pain","year":"2021","journal-title":"Vet. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1407","DOI":"10.1038\/s41598-018-38130-0","article-title":"A Human iPSC-derived 3D platform using primary brain cancer cells to study drug development and personalized medicine","volume":"9","author":"Plummer","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3991","DOI":"10.1038\/s41467-019-11867-6","article-title":"Patient-derived lung cancer organoids as in vitro cancer models for therapeutic screening","volume":"10","author":"Kim","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e1800268","DOI":"10.1002\/biot.201800268","article-title":"Establishment of 2D Cell Cultures Derived from 3D MCF-7 Spheroids Displaying a Doxorubicin Resistant Profile","volume":"14","author":"Nunes","year":"2019","journal-title":"Biotechnol. J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1089\/ten.tea.2017.0397","article-title":"Bioengineered Submucosal Organoids for In Vitro Modeling of Colorectal Cancer","volume":"23","author":"Devarasetty","year":"2017","journal-title":"Tissue Eng. Part A"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1021\/acs.chemrestox.9b00499","article-title":"Quantitative Systems Toxicology Modeling to Address Key Safety Questions in Drug Development: A Focus of the TransQST Consortium","volume":"33","author":"Ferreira","year":"2020","journal-title":"Chem. Res. Toxicol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.2165\/00003088-200544100-00005","article-title":"Pharmacokinetic drug interactions of gefitinib with rifampicin, itraconazole and metoprolol","volume":"44","author":"Swaisland","year":"2005","journal-title":"Clin. Pharmacokinet."},{"key":"ref_28","unstructured":"(2019, March 05). Astrazeneca, Available online: https:\/\/www.accessdata.fda.gov\/drugsatfda_docs\/label\/2018\/206995s003lbl.pdf."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1002\/jcph.1036","article-title":"Physiologically Based Pharmacokinetic Modeling to Evaluate the Systemic Exposure of Gefitinib in CYP2D6 Ultrarapid Metabolizers and Extensive Metabolizers","volume":"58","author":"Chen","year":"2018","journal-title":"J. Clin. Pharmacol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"726","DOI":"10.2105\/AJPH.86.5.726","article-title":"Adjusting for multiple testing when reporting research results: The Bonferroni vs Holm methods","volume":"86","author":"Aickin","year":"1996","journal-title":"Am. J. Public Health"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"724","DOI":"10.1002\/jcb.20678","article-title":"Gefitinib, a selective EGFR tyrosine kinase inhibitor, induces apoptosis through activation of Bax in human gallbladder adenocarcinoma cells","volume":"97","author":"Ariyama","year":"2006","journal-title":"J. Cell. Biochem."},{"key":"ref_32","first-page":"115","article-title":"Gefitinib-resistance is related to BIM expression in non-small cell lung cancer cell lines","volume":"28","author":"Li","year":"2013","journal-title":"Cancer Biother. Radiopharm."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.lfs.2014.10.008","article-title":"EGFR inhibitor-driven endoplasmic reticulum stress-mediated injury on intestinal epithelial cells","volume":"119","author":"Hong","year":"2014","journal-title":"Life Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.taap.2014.04.015","article-title":"Erlotinib promotes endoplasmic reticulum stress-mediated injury in the intestinal epithelium","volume":"278","author":"Fan","year":"2014","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1189","DOI":"10.1016\/j.cgh.2009.04.024","article-title":"A new mechanism for bile acid diarrhea: Defective feedback inhibition of bile acid biosynthesis","volume":"7","author":"Walters","year":"2009","journal-title":"Clin. Gastroenterol. Hepatol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"R113","DOI":"10.1016\/j.cub.2007.01.008","article-title":"FOXO transcription factors","volume":"17","author":"Carter","year":"2007","journal-title":"Curr. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Pottier, C., Fresnais, M., Gilon, M., Jerusalem, G., Longuespee, R., and Sounni, N.E. (2020). Tyrosine Kinase Inhibitors in Cancer: Breakthrough and Challenges of Targeted Therapy. Cancers, 12.","DOI":"10.3390\/cancers12030731"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1002\/9780470942390.mo130179","article-title":"Establishment of Gastrointestinal Epithelial Organoids","volume":"3","author":"Mahe","year":"2013","journal-title":"Curr. Protoc. Mouse Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1762","DOI":"10.1053\/j.gastro.2011.07.050","article-title":"Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett\u2019s epithelium","volume":"141","author":"Sato","year":"2011","journal-title":"Gastroenterology"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Rodrigues, D., Coyle, L., F\u00fczi, B., Ferreira, S., Jo, H., Herpers, B., Chung, S.-W., Fisher, C., Kleinjans, J.C.S., and Jennen, D. (2022). Unravelling Mechanisms of Doxorubicin-Induced Toxicity in 3D Human Intestinal Organoids. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23031286"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.dnarep.2008.12.001","article-title":"HCT116 cells deficient in p21(Waf1) are hypersensitive to tyrosine kinase inhibitors and adriamycin through a mechanism unrelated to p21 and dependent on p53","volume":"8","author":"Ferrandiz","year":"2009","journal-title":"DNA Repair."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1038\/mt.2009.226","article-title":"Amphiregulin promotes BAX inhibition and resistance to gefitinib in non-small-cell lung cancers","volume":"18","author":"Busser","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1860","DOI":"10.3892\/or.2015.3764","article-title":"AURKA upregulation plays a role in fibroblast-reduced gefitinib sensitivity in the NSCLC cell line HCC827","volume":"33","author":"Chen","year":"2015","journal-title":"Oncol. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1211\/jpp.61.05.0002","article-title":"Cytochrome P450-mediated metabolism in the human gut wall","volume":"61","author":"Thelen","year":"2009","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1879","DOI":"10.1124\/dmd.118.083428","article-title":"Regional Differences in Human Intestinal Drug Metabolism","volume":"46","author":"Iswandana","year":"2018","journal-title":"Drug Metab. Dispos."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"5483","DOI":"10.1021\/acs.biochem.6b00342","article-title":"Cholesterol Biosynthesis: A Mechanistic Overview","volume":"55","author":"Cerqueira","year":"2016","journal-title":"Biochemistry"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1038\/nrm.2017.95","article-title":"AMPK: Guardian of metabolism and mitochondrial homeostasis","volume":"19","author":"Herzig","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2316","DOI":"10.1002\/jcp.22570","article-title":"Lipid raft localization of EGFR alters the response of cancer cells to the EGFR tyrosine kinase inhibitor gefitinib","volume":"226","author":"Irwin","year":"2011","journal-title":"J. Cell. Physiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"6722","DOI":"10.1002\/jcp.26351","article-title":"High cholesterol in lipid rafts reduces the sensitivity to EGFR-TKI therapy in non-small cell lung cancer","volume":"233","author":"Chen","year":"2018","journal-title":"J. Cell. Physiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1096\/fba.2019-00081","article-title":"Lung cancer cells survive epidermal growth factor receptor tyrosine kinase inhibitor exposure through upregulation of cholesterol synthesis","volume":"2","author":"Howell","year":"2020","journal-title":"FASEB Bioadv."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1208\/s12248-009-9099-y","article-title":"Population-based mechanistic prediction of oral drug absorption","volume":"11","author":"Jamei","year":"2009","journal-title":"AAPS J."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1517\/17425250802691074","article-title":"The Simcyp population-based ADME simulator","volume":"5","author":"Jamei","year":"2009","journal-title":"Expert Opin. Drug Metab. Toxicol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.tiv.2018.12.017","article-title":"VIVD: Virtual in vitro distribution model for the mechanistic prediction of intracellular concentrations of chemicals in in vitro toxicity assays","volume":"58","author":"Fisher","year":"2019","journal-title":"Toxicol. In Vitro"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Di, Z., Klop, M.J., Rogkoti, V.M., Le Devedec, S.E., van de Water, B., Verbeek, F.J., Price, L.S., and Meerman, J.H. (2014). Ultra high content image analysis and phenotype profiling of 3D cultured micro-tissues. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0109688"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","article-title":"Trimmomatic: A flexible trimmer for Illumina sequence data","volume":"30","author":"Bolger","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_56","unstructured":"Andrews, S. (2021, April 21). FastQC A Quality Control Tool for High Throughput Sequence Data. Available online: http:\/\/www.bioinformatics.babraham.ac.uk\/projects\/fastqc\/."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"D793","DOI":"10.1093\/nar\/gks1055","article-title":"The ConsensusPathDB interaction database: 2013 update","volume":"41","author":"Kamburov","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"D649","DOI":"10.1093\/nar\/gkx1132","article-title":"The Reactome Pathway Knowledgebase","volume":"46","author":"Fabregat","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"D613","DOI":"10.1093\/nar\/gkaa1024","article-title":"WikiPathways: Connecting communities","volume":"49","author":"Martens","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"2441","DOI":"10.1039\/C4MB00287C","article-title":"KEGG-PATH: Kyoto encyclopedia of genes and genomes-based pathway analysis using a path analysis model","volume":"10","author":"Du","year":"2014","journal-title":"Mol. Biosyst."},{"key":"ref_62","unstructured":"(2021, June 26). BioRender. Available online: https:\/\/biorender.com\/."},{"key":"ref_63","unstructured":"Information NCfB (2021, November 30). PubChem Compound Summary for CID 123631, Gefitinib, Available online: https:\/\/pubchem.ncbi.nlm.nih.gov\/compound\/Gefitinib."},{"key":"ref_64","unstructured":"Meylan WM, H.P. (2000). User\u2019s Guide for HENRYWIN Version 3.10: Henry\u2019s Law Constant Program, Syracuse Research Corporation."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1007\/s10637-006-5269-2","article-title":"Binding of gefitinib, an inhibitor of epidermal growth factor receptor-tyrosine kinase, to plasma proteins and blood cells: In vitro and in cancer patients","volume":"24","author":"Li","year":"2006","journal-title":"Investig. New Drugs"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1002\/cpt.1672","article-title":"Evaluating the Role of Solubility in Oral Absorption of Poorly Water-Soluble Drugs Using Physiologically-Based Pharmacokinetic Modeling","volume":"107","author":"Fink","year":"2020","journal-title":"Clin. Pharmacol. Ther."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"1259","DOI":"10.1002\/jps.20322","article-title":"Physiologically based pharmacokinetic modeling 1: Predicting the tissue distribution of moderate-to-strong bases","volume":"94","author":"Rodgers","year":"2005","journal-title":"J. Pharm. Sci."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1238","DOI":"10.1002\/jps.20502","article-title":"Physiologically based pharmacokinetic modelling 2: Predicting the tissue distribution of acids, very weak bases, neutrals and zwitterions","volume":"95","author":"Rodgers","year":"2006","journal-title":"J. Pharm. Sci."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"669","DOI":"10.2165\/00003088-200847100-00004","article-title":"General framework for the prediction of oral drug interactions caused by CYP3A4 induction from in vivo information","volume":"47","author":"Ohno","year":"2008","journal-title":"Clin. Pharmacokinet."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.2147\/DDDT.S133814","article-title":"Functional characterization of wild-type and 24 CYP2D6 allelic variants on gefitinib metabolism in vitro","volume":"11","author":"Fang","year":"2017","journal-title":"Drug Des. Dev. Ther."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/4\/2213\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:21:23Z","timestamp":1760134883000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/4\/2213"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,17]]},"references-count":70,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["ijms23042213"],"URL":"https:\/\/doi.org\/10.3390\/ijms23042213","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,17]]}}}