Development of a novel AAK1 inhibitor via Kinobeads-based screening.

Autor: Yoshida A; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Ohtsuka S; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Matsumoto F; Department of Science Education, Graduate School of Education, Okayama University, Okayama, 700-8530, Japan., Miyagawa T; Department of Science Education, Graduate School of Education, Okayama University, Okayama, 700-8530, Japan., Okino R; Department of Science Education, Graduate School of Education, Okayama University, Okayama, 700-8530, Japan., Ikeda Y; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Tada N; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Gotoh A; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Magari M; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Hatano N; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Morishita R; CellFree Sciences Co. Ltd, Matsuyama, 790-8577, Japan., Satoh A; Organelle Systems Biotechnology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan., Sunatsuki Y; Graduate School of Natural Science and Technology, Okayama University, Okayama, 700-8530, Japan., Nilsson UJ; Department of Chemistry, Lund University, Box 124, 221 00, Lund, Sweden., Ishikawa T; Department of Science Education, Graduate School of Education, Okayama University, Okayama, 700-8530, Japan. teruhiko@cc.okayama-u.ac.jp., Tokumitsu H; Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan. tokumit@okayama-u.ac.jp.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Mar 20; Vol. 14 (1), pp. 6723. Date of Electronic Publication: 2024 Mar 20.
DOI: 10.1038/s41598-024-57051-9
Abstrakt: A chemical proteomics approach using Ca 2+ /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 50  = 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 (IC 50  = 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 50  = 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.
(© 2024. The Author(s).)
Databáze: MEDLINE
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