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ArticleActa histochemica et cytochemica2025

Immunohistochemical Study of the Localization of Ibrutinib-Protein Conjugates in the Rat Gastrointestinal Tract.

Hiroto Kataoka, Tetsuya Saita, Yutaro Yamamoto, Sakiko Kimura, Rintaro Sogawa, Chisato Shimanoe

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Article in Acta histochemica et cytochemica, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

6 authors.

Hiroto KataokaApplied Life Science Department, Faculty of Biotechnology and Life Science, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan.
Tetsuya SaitaApplied Life Science Department, Faculty of Biotechnology and Life Science, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan.
Yutaro YamamotoApplied Life Science Department, Faculty of Biotechnology and Life Science, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan.
Sakiko KimuraDepartment of Pharmacy, Saga University Hospital, 5-1-1 Nabeshima, Saga 849-8501, Japan.
Rintaro SogawaDepartment of Pharmacy, Saga University Hospital, 5-1-1 Nabeshima, Saga 849-8501, Japan.
Chisato ShimanoeDepartment of Pharmacy, Saga University Hospital, 5-1-1 Nabeshima, Saga 849-8501, Japan.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Ibrutinib is an oral irreversible Bruton's tyrosine kinase (BTK) inhibitor that blocks BTK activity by forming covalent bonds with the thiol group of cysteine in the ATP-binding pocket via Michael addition. However, it also reacts with a variety of off-target nonspecific proteins. In this study, we attempted to generate a specific antibody against ibrutinib and develop an immunohistochemical method to detect the ibrutinib-protein conjugates. Ibrutinib has the same amino group as the nucleobase adenine. Paraformaldehyde fixation could not fix it to the tissue via this amino group. However, ibrutinib covalently binds to proteins such as BTKs to exert its action and is therefore immobilized in tissue as ibrutinib-protein conjugates. Thus, immunohistochemistry for ibrutinib detects the location of the ibrutinib-protein conjugates, that is, the sites of covalently bound to the tissue via Michael addition. Using this immunohistochemical method, we visualized the ibrutinib-protein conjugates in the rat gastrointestinal tract (gastric body, duodenum, jejunum, ileum, and colon). This study is the first to elucidate the location of the ibrutinib-protein conjugates in the rat gastrointestinal tract and helps to clarify the mechanism of ibrutinib-induced toxicity.

Indexed as

gastrointestinal tractibrutinibimmunohistochemistrylocalization

Identifiers

PMID40535471
PMCPMC12173639

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