Development of a carbon-11 labeled thienopyrimidine based radioligand for EGFR-tyrosine-kinase-inhibitor (TKI) specific PET imaging of glioblastoma. Guttormsen, Y., Moldes-Anaya, A., Fjellaksel, R., Oteiza, A., Martin-Armas, M., Lindemann, M., Bihler, J., Hoff, B. H., Jakobsen, S., Sundby, E., Sundset, R., & Kranz, M. Journal of Nuclear Medicine, 62(supplement 1):1476–1476, Society of Nuclear Medicine, 2021. ![Development of a carbon-11 labeled thienopyrimidine based radioligand for EGFR-tyrosine-kinase-inhibitor (TKI) specific PET imaging of glioblastoma [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper abstract bibtex 1 download 1476Introduction: Glioblastoma (GBM) is a devastating cancer disease affecting 3 of 100000 Europeans. Furthermore, GBM accounts for 25% of all malignant tumors of the central nervous system. Combined approaches, including surgery, chemotherapy and external radiation have shown to only slightly prolong the survival of patients to a maximum of 14 months after first diagnosis. Reasons for this ineffective treatment are the tumor growth pattern, its invasive vascularization and high heterogenicity. Hence, a specific PET radioligand for visualization of GBM biomarkers like EGFR-TKI might help to improve diagnostic and therapy monitoring following anti-angiogenic treatment. Here, we describe the development of a thienopyrimidine based radiotracer from a library of EGFR-TKI from Bugge et al. [1] and its subsequent in vivo characterization and visualization. Methods: The non-radioactive thienopyrimidine standard 1 and the desmethyl precursor 2 were synthesized according to published methods [1]. Radiosynthesis of [11C]1 was achieved by methylation with [11C]MeI using NaOH as a base in DMSO at 100 °C for 3-5 min in a TRACERlab FX2C module. [11C]1 was purified by preparative HPLC and further passed through a C-18 Sep-Pak cartridge. The eluted ethanolic extract was evaporated to approximately 30 \textmul and reconstituted to a final volume of 400 \textmul with 1% polysorbate 80 in isotonic saline for in vivo application. The cell association was evaluated at room temperature by in vitro saturation studies in A431 cells using Erlotinib as a blocking agent. Radiotracer biodistribution in vivo was studied by 1 h dynamic simultaneous PET/MR imaging (DRYMAG 7024/PET, MRsolutions, Guildford, UK) following [11C]1 i.v. injection (5.8\textpm4.3 MBq) in healthy female CD1 mice (31.7\textpm4.2 g, n=4). Initial metabolism was studied in healthy CD1 mice (n=3) administered with 130\textpm45 MBq [11C]1. Animals were sacrificed five minutes after administration and brain, blood and liver were collected and homogenized in ACN/H2O (7:3). Tissue homogenates and plasma were further analyzed by HPLC-RAD-MS. Results: Compounds 1 and 2 were obtained in 29% and 39% overall yields over six steps. Radiomethylation of 2 with [11C]MeI resulted in [11C]1 with 8.9\textpm3.8% (n=12) radiochemical yield (non-decay corrected, EOB) in 45 min total synthesis time. The radiochemical purity was found to be >99% (n=12) and the molar activity achieved was 115\textpm80 GBq/\textmumol (n=12). In vitro stability in human serum shows >99% intact [11C]1 after 30 minutes. Compound [11C]1 showed uptake in EGFR overexpressing A431 cells which was displaceable with Erlotinib. The radiotracer showed blood-brain-barrier (BBB) penetration and nonsignificant amounts of radiometabolites were detected with the methods applied. However, possible hydroxylated metabolites were found in trace amounts by LC/MS analysis. PET/MR imaging revealed high brain uptake (SUV=1.4 at 5 min p.i.) followed by a wash-out during the investigation time. Conclusion: Our preliminary PET/MR and ex vivo metabolism results indicate that compound [11C]1 crosses the BBB and that non-significant radiometabolite amounts are detected with the analysis methods applied . Cellular uptake in A431 cells renders [11C]1 suitable for subsequent in vivo studies. Hence, further studies in brain tumor bearing animals are planned to show specific binding to GBM tissue and to further develop the radiotracer as a tool for brain tumor imaging with PET. 1. Bugge, S., et al., Structure-activity study leading to identification of a highly active thienopyrimidine based EGFR inhibitor. European Journal of Medicinal Chemistry, 2014. 75: p. 354-374. Acknowledgements: This study is financed by 180°N the Norwegian Nuclear Medicine Consortium
@article {Guttormsen1476,
author = {Guttormsen, Yngve and Moldes-Anaya, Angel and Fjellaksel, Richard and Oteiza, Ana and Martin-Armas, Montserrat and Lindemann, Marcel and Bihler, Julia and Hoff, Bard Helge and Jakobsen, Steen and Sundby, Eirik and Sundset, Rune and Kranz, Mathias},
title = {Development of a carbon-11 labeled thienopyrimidine based radioligand for EGFR-tyrosine-kinase-inhibitor (TKI) specific PET imaging of glioblastoma},
volume = {62},
number = {supplement 1},
pages = {1476--1476},
year = {2021},
publisher = {Society of Nuclear Medicine},
abstract = {1476Introduction: Glioblastoma (GBM) is a devastating cancer disease affecting 3 of 100000 Europeans. Furthermore, GBM accounts for 25\% of all malignant tumors of the central nervous system. Combined approaches, including surgery, chemotherapy and external radiation have shown to only slightly prolong the survival of patients to a maximum of 14 months after first diagnosis. Reasons for this ineffective treatment are the tumor growth pattern, its invasive vascularization and high heterogenicity. Hence, a specific PET radioligand for visualization of GBM biomarkers like EGFR-TKI might help to improve diagnostic and therapy monitoring following anti-angiogenic treatment. Here, we describe the development of a thienopyrimidine based radiotracer from a library of EGFR-TKI from Bugge et al. [1] and its subsequent in vivo characterization and visualization. Methods: The non-radioactive thienopyrimidine standard 1 and the desmethyl precursor 2 were synthesized according to published methods [1]. Radiosynthesis of [11C]1 was achieved by methylation with [11C]MeI using NaOH as a base in DMSO at 100 {\textdegree}C for 3-5 min in a TRACERlab FX2C module. [11C]1 was purified by preparative HPLC and further passed through a C-18 Sep-Pak cartridge. The eluted ethanolic extract was evaporated to approximately 30 {\textmu}l and reconstituted to a final volume of 400 {\textmu}l with 1\% polysorbate 80 in isotonic saline for in vivo application. The cell association was evaluated at room temperature by in vitro saturation studies in A431 cells using Erlotinib as a blocking agent. Radiotracer biodistribution in vivo was studied by 1 h dynamic simultaneous PET/MR imaging (DRYMAG 7024/PET, MRsolutions, Guildford, UK) following [11C]1 i.v. injection (5.8{\textpm}4.3 MBq) in healthy female CD1 mice (31.7{\textpm}4.2 g, n=4). Initial metabolism was studied in healthy CD1 mice (n=3) administered with 130{\textpm}45 MBq [11C]1. Animals were sacrificed five minutes after administration and brain, blood and liver were collected and homogenized in ACN/H2O (7:3). Tissue homogenates and plasma were further analyzed by HPLC-RAD-MS. Results: Compounds 1 and 2 were obtained in 29\% and 39\% overall yields over six steps. Radiomethylation of 2 with [11C]MeI resulted in [11C]1 with 8.9{\textpm}3.8\% (n=12) radiochemical yield (non-decay corrected, EOB) in 45 min total synthesis time. The radiochemical purity was found to be \>99\% (n=12) and the molar activity achieved was 115{\textpm}80 GBq/{\textmu}mol (n=12). In vitro stability in human serum shows \>99\% intact [11C]1 after 30 minutes. Compound [11C]1 showed uptake in EGFR overexpressing A431 cells which was displaceable with Erlotinib. The radiotracer showed blood-brain-barrier (BBB) penetration and nonsignificant amounts of radiometabolites were detected with the methods applied. However, possible hydroxylated metabolites were found in trace amounts by LC/MS analysis. PET/MR imaging revealed high brain uptake (SUV=1.4 at 5 min p.i.) followed by a wash-out during the investigation time. Conclusion: Our preliminary PET/MR and ex vivo metabolism results indicate that compound [11C]1 crosses the BBB and that non-significant radiometabolite amounts are detected with the analysis methods applied . Cellular uptake in A431 cells renders [11C]1 suitable for subsequent in vivo studies. Hence, further studies in brain tumor bearing animals are planned to show specific binding to GBM tissue and to further develop the radiotracer as a tool for brain tumor imaging with PET. 1. Bugge, S., et al., Structure-activity study leading to identification of a highly active thienopyrimidine based EGFR inhibitor. European Journal of Medicinal Chemistry, 2014. 75: p. 354-374. Acknowledgements: This study is financed by 180{\textdegree}N the Norwegian Nuclear Medicine Consortium},
issn = {0161-5505},
URL = {https://jnm.snmjournals.org/content/62/supplement_1/1476},
journal = {Journal of Nuclear Medicine}
}
Downloads: 1
{"_id":"oQGxNs4NcyfPEznqS","bibbaseid":"guttormsen-moldesanaya-fjellaksel-oteiza-martinarmas-lindemann-bihler-hoff-etal-developmentofacarbon11labeledthienopyrimidinebasedradioligandforegfrtyrosinekinaseinhibitortkispecificpetimagingofglioblastoma-2021","author_short":["Guttormsen, Y.","Moldes-Anaya, A.","Fjellaksel, R.","Oteiza, A.","Martin-Armas, M.","Lindemann, M.","Bihler, J.","Hoff, B. H.","Jakobsen, S.","Sundby, E.","Sundset, R.","Kranz, M."],"bibdata":{"bibtype":"article","type":"article","author":[{"propositions":[],"lastnames":["Guttormsen"],"firstnames":["Yngve"],"suffixes":[]},{"propositions":[],"lastnames":["Moldes-Anaya"],"firstnames":["Angel"],"suffixes":[]},{"propositions":[],"lastnames":["Fjellaksel"],"firstnames":["Richard"],"suffixes":[]},{"propositions":[],"lastnames":["Oteiza"],"firstnames":["Ana"],"suffixes":[]},{"propositions":[],"lastnames":["Martin-Armas"],"firstnames":["Montserrat"],"suffixes":[]},{"propositions":[],"lastnames":["Lindemann"],"firstnames":["Marcel"],"suffixes":[]},{"propositions":[],"lastnames":["Bihler"],"firstnames":["Julia"],"suffixes":[]},{"propositions":[],"lastnames":["Hoff"],"firstnames":["Bard","Helge"],"suffixes":[]},{"propositions":[],"lastnames":["Jakobsen"],"firstnames":["Steen"],"suffixes":[]},{"propositions":[],"lastnames":["Sundby"],"firstnames":["Eirik"],"suffixes":[]},{"propositions":[],"lastnames":["Sundset"],"firstnames":["Rune"],"suffixes":[]},{"propositions":[],"lastnames":["Kranz"],"firstnames":["Mathias"],"suffixes":[]}],"title":"Development of a carbon-11 labeled thienopyrimidine based radioligand for EGFR-tyrosine-kinase-inhibitor (TKI) specific PET imaging of glioblastoma","volume":"62","number":"supplement 1","pages":"1476–1476","year":"2021","publisher":"Society of Nuclear Medicine","abstract":"1476Introduction: Glioblastoma (GBM) is a devastating cancer disease affecting 3 of 100000 Europeans. Furthermore, GBM accounts for 25% of all malignant tumors of the central nervous system. Combined approaches, including surgery, chemotherapy and external radiation have shown to only slightly prolong the survival of patients to a maximum of 14 months after first diagnosis. Reasons for this ineffective treatment are the tumor growth pattern, its invasive vascularization and high heterogenicity. Hence, a specific PET radioligand for visualization of GBM biomarkers like EGFR-TKI might help to improve diagnostic and therapy monitoring following anti-angiogenic treatment. Here, we describe the development of a thienopyrimidine based radiotracer from a library of EGFR-TKI from Bugge et al. [1] and its subsequent in vivo characterization and visualization. Methods: The non-radioactive thienopyrimidine standard 1 and the desmethyl precursor 2 were synthesized according to published methods [1]. Radiosynthesis of [11C]1 was achieved by methylation with [11C]MeI using NaOH as a base in DMSO at 100 °C for 3-5 min in a TRACERlab FX2C module. [11C]1 was purified by preparative HPLC and further passed through a C-18 Sep-Pak cartridge. The eluted ethanolic extract was evaporated to approximately 30 \\textmul and reconstituted to a final volume of 400 \\textmul with 1% polysorbate 80 in isotonic saline for in vivo application. The cell association was evaluated at room temperature by in vitro saturation studies in A431 cells using Erlotinib as a blocking agent. Radiotracer biodistribution in vivo was studied by 1 h dynamic simultaneous PET/MR imaging (DRYMAG 7024/PET, MRsolutions, Guildford, UK) following [11C]1 i.v. injection (5.8\\textpm4.3 MBq) in healthy female CD1 mice (31.7\\textpm4.2 g, n=4). Initial metabolism was studied in healthy CD1 mice (n=3) administered with 130\\textpm45 MBq [11C]1. Animals were sacrificed five minutes after administration and brain, blood and liver were collected and homogenized in ACN/H2O (7:3). Tissue homogenates and plasma were further analyzed by HPLC-RAD-MS. Results: Compounds 1 and 2 were obtained in 29% and 39% overall yields over six steps. Radiomethylation of 2 with [11C]MeI resulted in [11C]1 with 8.9\\textpm3.8% (n=12) radiochemical yield (non-decay corrected, EOB) in 45 min total synthesis time. The radiochemical purity was found to be >99% (n=12) and the molar activity achieved was 115\\textpm80 GBq/\\textmumol (n=12). In vitro stability in human serum shows >99% intact [11C]1 after 30 minutes. Compound [11C]1 showed uptake in EGFR overexpressing A431 cells which was displaceable with Erlotinib. The radiotracer showed blood-brain-barrier (BBB) penetration and nonsignificant amounts of radiometabolites were detected with the methods applied. However, possible hydroxylated metabolites were found in trace amounts by LC/MS analysis. PET/MR imaging revealed high brain uptake (SUV=1.4 at 5 min p.i.) followed by a wash-out during the investigation time. Conclusion: Our preliminary PET/MR and ex vivo metabolism results indicate that compound [11C]1 crosses the BBB and that non-significant radiometabolite amounts are detected with the analysis methods applied . Cellular uptake in A431 cells renders [11C]1 suitable for subsequent in vivo studies. Hence, further studies in brain tumor bearing animals are planned to show specific binding to GBM tissue and to further develop the radiotracer as a tool for brain tumor imaging with PET. 1. Bugge, S., et al., Structure-activity study leading to identification of a highly active thienopyrimidine based EGFR inhibitor. European Journal of Medicinal Chemistry, 2014. 75: p. 354-374. Acknowledgements: This study is financed by 180°N the Norwegian Nuclear Medicine Consortium","issn":"0161-5505","url":"https://jnm.snmjournals.org/content/62/supplement_1/1476","journal":"Journal of Nuclear Medicine","bibtex":"@article {Guttormsen1476,\n\tauthor = {Guttormsen, Yngve and Moldes-Anaya, Angel and Fjellaksel, Richard and Oteiza, Ana and Martin-Armas, Montserrat and Lindemann, Marcel and Bihler, Julia and Hoff, Bard Helge and Jakobsen, Steen and Sundby, Eirik and Sundset, Rune and Kranz, Mathias},\n\ttitle = {Development of a carbon-11 labeled thienopyrimidine based radioligand for EGFR-tyrosine-kinase-inhibitor (TKI) specific PET imaging of glioblastoma},\n\tvolume = {62},\n\tnumber = {supplement 1},\n\tpages = {1476--1476},\n\tyear = {2021},\n\tpublisher = {Society of Nuclear Medicine},\n\tabstract = {1476Introduction: Glioblastoma (GBM) is a devastating cancer disease affecting 3 of 100000 Europeans. Furthermore, GBM accounts for 25\\% of all malignant tumors of the central nervous system. Combined approaches, including surgery, chemotherapy and external radiation have shown to only slightly prolong the survival of patients to a maximum of 14 months after first diagnosis. Reasons for this ineffective treatment are the tumor growth pattern, its invasive vascularization and high heterogenicity. Hence, a specific PET radioligand for visualization of GBM biomarkers like EGFR-TKI might help to improve diagnostic and therapy monitoring following anti-angiogenic treatment. Here, we describe the development of a thienopyrimidine based radiotracer from a library of EGFR-TKI from Bugge et al. [1] and its subsequent in vivo characterization and visualization. Methods: The non-radioactive thienopyrimidine standard 1 and the desmethyl precursor 2 were synthesized according to published methods [1]. Radiosynthesis of [11C]1 was achieved by methylation with [11C]MeI using NaOH as a base in DMSO at 100 {\\textdegree}C for 3-5 min in a TRACERlab FX2C module. [11C]1 was purified by preparative HPLC and further passed through a C-18 Sep-Pak cartridge. The eluted ethanolic extract was evaporated to approximately 30 {\\textmu}l and reconstituted to a final volume of 400 {\\textmu}l with 1\\% polysorbate 80 in isotonic saline for in vivo application. The cell association was evaluated at room temperature by in vitro saturation studies in A431 cells using Erlotinib as a blocking agent. Radiotracer biodistribution in vivo was studied by 1 h dynamic simultaneous PET/MR imaging (DRYMAG 7024/PET, MRsolutions, Guildford, UK) following [11C]1 i.v. injection (5.8{\\textpm}4.3 MBq) in healthy female CD1 mice (31.7{\\textpm}4.2 g, n=4). Initial metabolism was studied in healthy CD1 mice (n=3) administered with 130{\\textpm}45 MBq [11C]1. Animals were sacrificed five minutes after administration and brain, blood and liver were collected and homogenized in ACN/H2O (7:3). Tissue homogenates and plasma were further analyzed by HPLC-RAD-MS. Results: Compounds 1 and 2 were obtained in 29\\% and 39\\% overall yields over six steps. Radiomethylation of 2 with [11C]MeI resulted in [11C]1 with 8.9{\\textpm}3.8\\% (n=12) radiochemical yield (non-decay corrected, EOB) in 45 min total synthesis time. The radiochemical purity was found to be \\>99\\% (n=12) and the molar activity achieved was 115{\\textpm}80 GBq/{\\textmu}mol (n=12). In vitro stability in human serum shows \\>99\\% intact [11C]1 after 30 minutes. Compound [11C]1 showed uptake in EGFR overexpressing A431 cells which was displaceable with Erlotinib. The radiotracer showed blood-brain-barrier (BBB) penetration and nonsignificant amounts of radiometabolites were detected with the methods applied. However, possible hydroxylated metabolites were found in trace amounts by LC/MS analysis. PET/MR imaging revealed high brain uptake (SUV=1.4 at 5 min p.i.) followed by a wash-out during the investigation time. Conclusion: Our preliminary PET/MR and ex vivo metabolism results indicate that compound [11C]1 crosses the BBB and that non-significant radiometabolite amounts are detected with the analysis methods applied . Cellular uptake in A431 cells renders [11C]1 suitable for subsequent in vivo studies. Hence, further studies in brain tumor bearing animals are planned to show specific binding to GBM tissue and to further develop the radiotracer as a tool for brain tumor imaging with PET. 1. Bugge, S., et al., Structure-activity study leading to identification of a highly active thienopyrimidine based EGFR inhibitor. European Journal of Medicinal Chemistry, 2014. 75: p. 354-374. 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