CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil
Target engagement is a critical factor for therapeutic efficacy. Assessment of compound binding to native target proteins in live cells is therefore highly desirable in all stages of drug discovery. We report here the first compound library screen based on biophysical measurements of intracellular t...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2018
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/89820 http://hdl.handle.net/10220/47161 |
| _version_ | 1824455811517120512 |
|---|---|
| author | Almqvist, Helena Axelsson, Hanna Jafari, Rozbeh Dan, Chen Mateus, André Haraldsson, Martin Larsson, Andreas Molina, Daniel Martinez Artursson, Per Lundbäck, Thomas Nordlund, Pär |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Almqvist, Helena Axelsson, Hanna Jafari, Rozbeh Dan, Chen Mateus, André Haraldsson, Martin Larsson, Andreas Molina, Daniel Martinez Artursson, Per Lundbäck, Thomas Nordlund, Pär |
| author_sort | Almqvist, Helena |
| collection | NTU |
| description | Target engagement is a critical factor for therapeutic efficacy. Assessment of compound binding to native target proteins in live cells is therefore highly desirable in all stages of drug discovery. We report here the first compound library screen based on biophysical measurements of intracellular target binding, exemplified by human thymidylate synthase (TS). The screen selected accurately for all the tested known drugs acting on TS. We also identified TS inhibitors with novel chemistry and marketed drugs that were not previously known to target TS, including the DNA methyltransferase inhibitor decitabine. By following the cellular uptake and enzymatic conversion of known drugs we correlated the appearance of active metabolites over time with intracellular target engagement. These data distinguished a much slower activation of 5-fluorouracil when compared with nucleoside-based drugs. The approach establishes efficient means to associate drug uptake and activation with target binding during drug discovery. |
| first_indexed | 2025-02-19T03:44:08Z |
| format | Journal Article |
| id | ntu-10356/89820 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2025-02-19T03:44:08Z |
| publishDate | 2018 |
| record_format | dspace |
| spelling | ntu-10356/898202023-02-28T17:02:18Z CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil Almqvist, Helena Axelsson, Hanna Jafari, Rozbeh Dan, Chen Mateus, André Haraldsson, Martin Larsson, Andreas Molina, Daniel Martinez Artursson, Per Lundbäck, Thomas Nordlund, Pär School of Biological Sciences Screening DRNTU::Science::Biological sciences Molecular Biophysics Target engagement is a critical factor for therapeutic efficacy. Assessment of compound binding to native target proteins in live cells is therefore highly desirable in all stages of drug discovery. We report here the first compound library screen based on biophysical measurements of intracellular target binding, exemplified by human thymidylate synthase (TS). The screen selected accurately for all the tested known drugs acting on TS. We also identified TS inhibitors with novel chemistry and marketed drugs that were not previously known to target TS, including the DNA methyltransferase inhibitor decitabine. By following the cellular uptake and enzymatic conversion of known drugs we correlated the appearance of active metabolites over time with intracellular target engagement. These data distinguished a much slower activation of 5-fluorouracil when compared with nucleoside-based drugs. The approach establishes efficient means to associate drug uptake and activation with target binding during drug discovery. Published version 2018-12-21T04:25:04Z 2019-12-06T17:34:14Z 2018-12-21T04:25:04Z 2019-12-06T17:34:14Z 2016 Journal Article Almqvist, H., Axelsson, H., Jafari, R., Dan, C., Mateus, A., Haraldsson, M., . . . Nordlund, P. (2016). CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil. Nature Communications, 7, 11040-. doi:10.1038/ncomms11040 https://hdl.handle.net/10356/89820 http://hdl.handle.net/10220/47161 10.1038/ncomms11040 27010513 en Nature Communications © 2016 The Author(s) (Published by Nature Publishing Group). This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. 11 p. application/pdf |
| spellingShingle | Screening DRNTU::Science::Biological sciences Molecular Biophysics Almqvist, Helena Axelsson, Hanna Jafari, Rozbeh Dan, Chen Mateus, André Haraldsson, Martin Larsson, Andreas Molina, Daniel Martinez Artursson, Per Lundbäck, Thomas Nordlund, Pär CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil |
| title | CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil |
| title_full | CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil |
| title_fullStr | CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil |
| title_full_unstemmed | CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil |
| title_short | CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil |
| title_sort | cetsa screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5 fluorouracil |
| topic | Screening DRNTU::Science::Biological sciences Molecular Biophysics |
| url | https://hdl.handle.net/10356/89820 http://hdl.handle.net/10220/47161 |
| work_keys_str_mv | AT almqvisthelena cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT axelssonhanna cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT jafarirozbeh cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT danchen cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT mateusandre cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT haraldssonmartin cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT larssonandreas cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT molinadanielmartinez cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT arturssonper cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT lundbackthomas cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil AT nordlundpar cetsascreeningidentifiesknownandnovelthymidylatesynthaseinhibitorsandslowintracellularactivationof5fluorouracil |