TY - JOUR
T1 - Bicyclobutane carboxylic amide as a cysteine-directed strained electrophile for selective targeting of proteins
AU - Shindo, Naoya
AU - Ojida, Akio
AU - Tokunaga, Keisuke
AU - Sato, Mami
AU - Kuwata, Keiko
AU - Miura, Chizuru
AU - Fuchida, Hirokazu
AU - Matsunaga, Naoya
AU - Koyanagi, Satoru
AU - Ohdo, Shigehiro
N1 - Funding Information:
This work was supported by a Grant-in-Aid for Scientific Research on Innovative Areas “Chemistry for Multimolecular Crowding Biosystems” (JSPS KAKENHI grant no. JP17H06349) and Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) from AMED under grant number JP18am0101091. N.S. acknowledges Grant-in-Aid for Young Scientists B (JSPS KAKENHI grant no. JP17K15483), Grant-in-Aid for Scientific Research B (JSPS KAKENHI grant no. 19H02854), and AMED under grant no. JP20ak0101121 for their financial support. H.F. acknowledges JSPS Research Fellowships for Young Scientists. ITbM is supported by the World Premier International Research Center Initiative, Japan. K.K. acknowledges a Grant-in-Aid for Scientific Research on Innovative Areas (JSPS KAKENHI grant no. JP15H05955).
Publisher Copyright:
© 2020 American Chemical Society.
PY - 2020/10/28
Y1 - 2020/10/28
N2 - Expanding the repertoire of electrophiles with unique reactivity features would facilitate the development of covalent inhibitors with desirable reactivity profiles. We herein introduce bicyclo[1.1.0]butane (BCB) carboxylic amide as a new class of thiol-reactive electrophiles for selective and irreversible inhibition of targeted proteins. We first streamlined the synthetic routes to generate a variety of BCB amides. The strain-driven nucleophilic addition to BCB amides proceeded chemoselectively with cysteine thiols under neutral aqueous conditions, the rate of which was significantly slower than that of acrylamide. This reactivity profile of BCB amide was successfully exploited to develop covalent ligands targeting Bruton's tyrosine kinase (BTK). By tuning BCB amide reactivity and optimizing its disposition on the ligand, we obtained a selective covalent inhibitor of BTK. The in-gel activitybased protein profiling and mass spectrometry-based chemical proteomics revealed that the selected BCB amide had a higher target selectivity for BTK in human cells than did a Michael acceptor probe. Further chemical proteomic study revealed that BTK probes bearing different classes of electrophiles exhibited distinct off-target profiles. This result suggests that incorporation of BCB amide as a cysteine-directed electrophile could expand the capability to develop covalent inhibitors with the desired proteome reactivity profile.
AB - Expanding the repertoire of electrophiles with unique reactivity features would facilitate the development of covalent inhibitors with desirable reactivity profiles. We herein introduce bicyclo[1.1.0]butane (BCB) carboxylic amide as a new class of thiol-reactive electrophiles for selective and irreversible inhibition of targeted proteins. We first streamlined the synthetic routes to generate a variety of BCB amides. The strain-driven nucleophilic addition to BCB amides proceeded chemoselectively with cysteine thiols under neutral aqueous conditions, the rate of which was significantly slower than that of acrylamide. This reactivity profile of BCB amide was successfully exploited to develop covalent ligands targeting Bruton's tyrosine kinase (BTK). By tuning BCB amide reactivity and optimizing its disposition on the ligand, we obtained a selective covalent inhibitor of BTK. The in-gel activitybased protein profiling and mass spectrometry-based chemical proteomics revealed that the selected BCB amide had a higher target selectivity for BTK in human cells than did a Michael acceptor probe. Further chemical proteomic study revealed that BTK probes bearing different classes of electrophiles exhibited distinct off-target profiles. This result suggests that incorporation of BCB amide as a cysteine-directed electrophile could expand the capability to develop covalent inhibitors with the desired proteome reactivity profile.
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U2 - 10.1021/jacs.0c07490
DO - 10.1021/jacs.0c07490
M3 - Article
C2 - 33047956
AN - SCOPUS:85094568418
VL - 142
SP - 18522
EP - 18531
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 43
ER -