Fully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imaging
Introduction Indoleamine 2,3-dioxygenase (IDO) catalyzes the initial step in the catabolism of l-tryptophan along the kynurenine pathway and exerts immunosuppressive properties in inflammatory and tumor tissues by blocking locally T-lymphocyte proliferation. Recently, 1-(2-[19F]fluoroethyl)-dl-trypt...
| Main Authors: | , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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Elsevier
2016
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| _version_ | 1826260379451260928 |
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| author | Henrottin, J Lemaire, C Egrise, D Zervosen, A Van Den Eynde, B Plenevaux, A Franci, X Goldman, S Luxen, A |
| author_facet | Henrottin, J Lemaire, C Egrise, D Zervosen, A Van Den Eynde, B Plenevaux, A Franci, X Goldman, S Luxen, A |
| author_sort | Henrottin, J |
| collection | OXFORD |
| description | Introduction Indoleamine 2,3-dioxygenase (IDO) catalyzes the initial step in the catabolism of l-tryptophan along the kynurenine pathway and exerts immunosuppressive properties in inflammatory and tumor tissues by blocking locally T-lymphocyte proliferation. Recently, 1-(2-[19F]fluoroethyl)-dl-tryptophan (1-[19F]FE-dl-Trp) was reported as a good and specific substrate of this enzyme. Herein, the radiosynthesis of its radioactive isotopomer (1-[18F]FE-dl-Trp, dl-[18F]5) is presented along with in vitro enzymatic and cellular uptake studies. Methods The one-pot n.c.a. radiosynthesis of this novel potential PET imaging tracer, including HPLC purification and formulation, has been fully automated on a FASTlab™ synthesizer. Chiral separation of both isomers and their formulation were implemented on a second cassette. In vitro enzymatic and cellular uptake studies were then conducted with the d-, l- and dl-radiotracers. Results The radiolabeling of the tosylate precursor was performed in DMF (in 5 min; RCY: 57% (d.c.), n = 3). After hydrolysis, HPLC purification and formulation, dl-[18F]5 was obtained with a global radiochemical yield of 18 ± 3% (not decay corrected, n = 7, in 80 min) and a specific activity of 600 ± 180 GBq/μmol (n = 5). The subsequent separation of l- and d-enantiomers was performed by chiral HPLC and both were obtained after formulation with an RCY (d.c.) of 6.1% and 5.8%, respectively. In vitro enzymatic assays reveal that l-[18F]5 is a better substrate than d-[18F]5 for human IDO. In vitro cellular assays show an IDO-specific uptake of the racemate varying from 30% to 50% of that of l-[18F]5, and a negligible uptake of d-[18F]5. Conclusion In vitro studies show that l-[18F]5 is a good and specific substrate of hIDO, while presenting a very low efflux. These results confirm that l-[18F]5 could be a very useful PET radiotracer for IDO expressing cells in cancer imaging. |
| first_indexed | 2024-03-06T19:04:42Z |
| format | Journal article |
| id | oxford-uuid:14be154d-fc89-4f77-96c9-8f1cd16e213b |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:04:42Z |
| publishDate | 2016 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:14be154d-fc89-4f77-96c9-8f1cd16e213b2022-03-26T10:21:29ZFully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imagingJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:14be154d-fc89-4f77-96c9-8f1cd16e213bEnglishSymplectic Elements at OxfordElsevier2016Henrottin, JLemaire, CEgrise, DZervosen, AVan Den Eynde, BPlenevaux, AFranci, XGoldman, SLuxen, AIntroduction Indoleamine 2,3-dioxygenase (IDO) catalyzes the initial step in the catabolism of l-tryptophan along the kynurenine pathway and exerts immunosuppressive properties in inflammatory and tumor tissues by blocking locally T-lymphocyte proliferation. Recently, 1-(2-[19F]fluoroethyl)-dl-tryptophan (1-[19F]FE-dl-Trp) was reported as a good and specific substrate of this enzyme. Herein, the radiosynthesis of its radioactive isotopomer (1-[18F]FE-dl-Trp, dl-[18F]5) is presented along with in vitro enzymatic and cellular uptake studies. Methods The one-pot n.c.a. radiosynthesis of this novel potential PET imaging tracer, including HPLC purification and formulation, has been fully automated on a FASTlab™ synthesizer. Chiral separation of both isomers and their formulation were implemented on a second cassette. In vitro enzymatic and cellular uptake studies were then conducted with the d-, l- and dl-radiotracers. Results The radiolabeling of the tosylate precursor was performed in DMF (in 5 min; RCY: 57% (d.c.), n = 3). After hydrolysis, HPLC purification and formulation, dl-[18F]5 was obtained with a global radiochemical yield of 18 ± 3% (not decay corrected, n = 7, in 80 min) and a specific activity of 600 ± 180 GBq/μmol (n = 5). The subsequent separation of l- and d-enantiomers was performed by chiral HPLC and both were obtained after formulation with an RCY (d.c.) of 6.1% and 5.8%, respectively. In vitro enzymatic assays reveal that l-[18F]5 is a better substrate than d-[18F]5 for human IDO. In vitro cellular assays show an IDO-specific uptake of the racemate varying from 30% to 50% of that of l-[18F]5, and a negligible uptake of d-[18F]5. Conclusion In vitro studies show that l-[18F]5 is a good and specific substrate of hIDO, while presenting a very low efflux. These results confirm that l-[18F]5 could be a very useful PET radiotracer for IDO expressing cells in cancer imaging. |
| spellingShingle | Henrottin, J Lemaire, C Egrise, D Zervosen, A Van Den Eynde, B Plenevaux, A Franci, X Goldman, S Luxen, A Fully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imaging |
| title | Fully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imaging |
| title_full | Fully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imaging |
| title_fullStr | Fully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imaging |
| title_full_unstemmed | Fully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imaging |
| title_short | Fully automated radiosynthesis of N1-[18F]fluoroethyl-tryptophan and study of its biological activity as a new potential substrate for indoleamine 2,3-dioxygenase PET imaging |
| title_sort | fully automated radiosynthesis of n1 18f fluoroethyl tryptophan and study of its biological activity as a new potential substrate for indoleamine 2 3 dioxygenase pet imaging |
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