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Development of a novel AAK1 inhibitor via Kinobeads-based screening

Yoshida, Akari ; Ohtsuka, Satomi ; Matsumoto, Fumiya ; Miyagawa, Tomoyuki ; Okino, Rei ; Ikeda, Yumeya ; Tada, Natsume ; Gotoh, Akira ; Magari, Masaki and Hatano, Naoya , et al. (2024) In Scientific Reports 14. p.6723-6723
Abstract
A chemical proteomics approach using Ca2+/calmodulin-dependent protein kinase kinase (CaMKK) inhibitor–immobilized sepharose (TIM-063-Kinobeads) identified main targets such as CaMKKα/1 and β/2, and potential off-target kinases, including AP2-associated protein kinase 1 (AAK1), as TIM-063 interactants. Because TIM-063 interacted with the AAK1 catalytic domain and inhibited its enzymatic activity moderately (IC50 = 8.51 µM), we attempted to identify potential AAK1 inhibitors from TIM-063-derivatives and found a novel AAK1 inhibitor, TIM-098a (11-amino-2-hydroxy-7H-benzo[de]benzo[4,5]imidazo[2,1-a]isoquinolin-7-one) which is more potent (IC50 = 0.24 µM) than TIM-063 without any inhibitory... (More)
A chemical proteomics approach using Ca2+/calmodulin-dependent protein kinase kinase (CaMKK) inhibitor–immobilized sepharose (TIM-063-Kinobeads) identified main targets such as CaMKKα/1 and β/2, and potential off-target kinases, including AP2-associated protein kinase 1 (AAK1), as TIM-063 interactants. Because TIM-063 interacted with the AAK1 catalytic domain and inhibited its enzymatic activity moderately (IC50 = 8.51 µM), we attempted to identify potential AAK1 inhibitors from TIM-063-derivatives and found a novel AAK1 inhibitor, TIM-098a (11-amino-2-hydroxy-7H-benzo[de]benzo[4,5]imidazo[2,1-a]isoquinolin-7-one) which is more potent (IC50 = 0.24 µM) than TIM-063 without any inhibitory activity against CaMKK isoforms and a relative AAK1-selectivity among the Numb-associated kinases family. TIM-098a could inhibit AAK1 activity in transfected cultured cells (IC50 = 0.87 µM), indicating cell-membrane permeability of the compound. Overexpression of AAK1 in HeLa cells significantly reduced the number of early endosomes, which was blocked by treatment with 10 µM TIM-098a. These results indicate TIM-063-Kinobeads-based chemical proteomics is efficient for identifying off-target kinases and re-evaluating the kinase inhibitor (TIM-063), leading to the successful development of a novel inhibitory compound (TIM-098a) for AAK1, which could be a molecular probe for AAK1. TIM-098a may be a promising lead compound for a more potent, selective and therapeutically useful AAK1 inhibitor. (Less)
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Scientific Reports
volume
14
pages
12 pages
publisher
Nature Publishing Group
external identifiers
  • scopus:85188234504
  • pmid:38509168
ISSN
2045-2322
DOI
10.1038/s41598-024-57051-9
language
English
LU publication?
yes
additional info
© 2024. The Author(s).
id
338fd61a-6553-4d7d-92df-967d41c6c2d2
date added to LUP
2024-03-21 16:13:03
date last changed
2024-11-15 11:32:03
@article{338fd61a-6553-4d7d-92df-967d41c6c2d2,
  abstract     = {{A chemical proteomics approach using Ca<sup>2+</sup>/calmodulin-dependent protein kinase kinase (CaMKK) inhibitor–immobilized sepharose (TIM-063-Kinobeads) identified main targets such as CaMKKα/1 and β/2, and potential off-target kinases, including AP2-associated protein kinase 1 (AAK1), as TIM-063 interactants. Because TIM-063 interacted with the AAK1 catalytic domain and inhibited its enzymatic activity moderately (IC<sub>50</sub> = 8.51 µM), we attempted to identify potential AAK1 inhibitors from TIM-063-derivatives and found a novel AAK1 inhibitor, TIM-098a (11-amino-2-hydroxy-7<i>H</i>-benzo[de]benzo[4,5]imidazo[2,1-<i>a</i>]isoquinolin-7-one) which is more potent (IC<sub>50</sub> = 0.24 µM) than TIM-063 without any inhibitory activity against CaMKK isoforms and a relative AAK1-selectivity among the Numb-associated kinases family. TIM-098a could inhibit AAK1 activity in transfected cultured cells (IC<sub>50</sub> = 0.87 µM), indicating cell-membrane permeability of the compound. Overexpression of AAK1 in HeLa cells significantly reduced the number of early endosomes, which was blocked by treatment with 10 µM TIM-098a. These results indicate TIM-063-Kinobeads-based chemical proteomics is efficient for identifying off-target kinases and re-evaluating the kinase inhibitor (TIM-063), leading to the successful development of a novel inhibitory compound (TIM-098a) for AAK1, which could be a molecular probe for AAK1. TIM-098a may be a promising lead compound for a more potent, selective and therapeutically useful AAK1 inhibitor.}},
  author       = {{Yoshida, Akari and Ohtsuka, Satomi and Matsumoto, Fumiya and Miyagawa, Tomoyuki and Okino, Rei and Ikeda, Yumeya and Tada, Natsume and Gotoh, Akira and Magari, Masaki and Hatano, Naoya and Morishita, Ryo and Satoh, Ayano and Sunatsuki, Yukinari and Nilsson, Ulf J. and Ishikawa, Teruhiko and Tokumitsu, Hiroshi}},
  issn         = {{2045-2322}},
  language     = {{eng}},
  month        = {{03}},
  pages        = {{6723--6723}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Scientific Reports}},
  title        = {{Development of a novel AAK1 inhibitor via Kinobeads-based screening}},
  url          = {{http://dx.doi.org/10.1038/s41598-024-57051-9}},
  doi          = {{10.1038/s41598-024-57051-9}},
  volume       = {{14}},
  year         = {{2024}},
}