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CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens
The proof of target engagement (TE) is a key element for evaluating potential investment in drug development. The cellular thermal shift assay (CETSA) is expected to facilitate direct measurement of intracellular TE at all stages of drug development. However, there have been no reports of applying t...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5638916/ https://www.ncbi.nlm.nih.gov/pubmed/29026104 http://dx.doi.org/10.1038/s41598-017-12513-1 |
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author | Ishii, Tsuyoshi Okai, Takuro Iwatani-Yoshihara, Misa Mochizuki, Manabu Unno, Satoko Kuno, Masako Yoshikawa, Masato Shibata, Sachio Nakakariya, Masanori Yogo, Takatoshi Kawamoto, Tomohiro |
author_facet | Ishii, Tsuyoshi Okai, Takuro Iwatani-Yoshihara, Misa Mochizuki, Manabu Unno, Satoko Kuno, Masako Yoshikawa, Masato Shibata, Sachio Nakakariya, Masanori Yogo, Takatoshi Kawamoto, Tomohiro |
author_sort | Ishii, Tsuyoshi |
collection | PubMed |
description | The proof of target engagement (TE) is a key element for evaluating potential investment in drug development. The cellular thermal shift assay (CETSA) is expected to facilitate direct measurement of intracellular TE at all stages of drug development. However, there have been no reports of applying this technology to comprehensive animal and clinical studies. This report demonstrates that CETSA can not only quantitatively evaluate the drug-TE in mouse peripheral blood, but also confirm TE in animal tissues exemplified by using the receptor interacting protein 1 kinase (RIPK1) lead compound we have developed. Our established semi-automated system allows evaluation of the structure-activity relationship using native RIPK1 in culture cell lines, and also enables estimation of drug occupancy ratio in mouse peripheral blood mononuclear cells. Moreover, optimized tissue homogenisation enables monitoring of the in vivo drug-TE in spleen and brain. Our results indicate that CETSA methodology will provide an efficient tool for preclinical and clinical drug development. |
format | Online Article Text |
id | pubmed-5638916 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-56389162017-10-18 CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens Ishii, Tsuyoshi Okai, Takuro Iwatani-Yoshihara, Misa Mochizuki, Manabu Unno, Satoko Kuno, Masako Yoshikawa, Masato Shibata, Sachio Nakakariya, Masanori Yogo, Takatoshi Kawamoto, Tomohiro Sci Rep Article The proof of target engagement (TE) is a key element for evaluating potential investment in drug development. The cellular thermal shift assay (CETSA) is expected to facilitate direct measurement of intracellular TE at all stages of drug development. However, there have been no reports of applying this technology to comprehensive animal and clinical studies. This report demonstrates that CETSA can not only quantitatively evaluate the drug-TE in mouse peripheral blood, but also confirm TE in animal tissues exemplified by using the receptor interacting protein 1 kinase (RIPK1) lead compound we have developed. Our established semi-automated system allows evaluation of the structure-activity relationship using native RIPK1 in culture cell lines, and also enables estimation of drug occupancy ratio in mouse peripheral blood mononuclear cells. Moreover, optimized tissue homogenisation enables monitoring of the in vivo drug-TE in spleen and brain. Our results indicate that CETSA methodology will provide an efficient tool for preclinical and clinical drug development. Nature Publishing Group UK 2017-10-12 /pmc/articles/PMC5638916/ /pubmed/29026104 http://dx.doi.org/10.1038/s41598-017-12513-1 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Ishii, Tsuyoshi Okai, Takuro Iwatani-Yoshihara, Misa Mochizuki, Manabu Unno, Satoko Kuno, Masako Yoshikawa, Masato Shibata, Sachio Nakakariya, Masanori Yogo, Takatoshi Kawamoto, Tomohiro CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens |
title | CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens |
title_full | CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens |
title_fullStr | CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens |
title_full_unstemmed | CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens |
title_short | CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens |
title_sort | cetsa quantitatively verifies in vivo target engagement of novel ripk1 inhibitors in various biospecimens |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5638916/ https://www.ncbi.nlm.nih.gov/pubmed/29026104 http://dx.doi.org/10.1038/s41598-017-12513-1 |
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