Enhanced Suppression of a Protein-Protein Interaction in Cells Using Small-Molecule Covalent Inhibitors Based on an N-Acyl- N-alkyl Sulfonamide Warhead

Tsuyoshi Ueda; Tomonori Tamura; Masaharu Kawano; Keiya Shiono; Fruzsina Hobor; Andrew J. Wilson; Itaru Hamachi

doi:10.1021/jacs.1c00703

Enhanced Suppression of a Protein-Protein Interaction in Cells Using Small-Molecule Covalent Inhibitors Based on an N-Acyl- N-alkyl Sulfonamide Warhead

Tsuyoshi Ueda, Tomonori Tamura^*, Masaharu Kawano, Keiya Shiono, Fruzsina Hobor, Andrew J. Wilson, Itaru Hamachi^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

26 Citations (Scopus)

Abstract

Protein-protein interactions (PPIs) intimately govern various biological processes and disease states and therefore have been identified as attractive therapeutic targets for small-molecule drug discovery. However, the development of highly potent inhibitors for PPIs has proven to be extremely challenging with limited clinical success stories. Herein, we report irreversible inhibitors of the human double minute 2 (HDM2)/p53 PPI, which employ a reactive N-acyl-N-alkyl sulfonamide (NASA) group as a warhead. Mass-based analysis successfully revealed the kinetics of covalent inhibition and the modification sites on HDM2 to be the N-terminal α-amine and Tyr67, both rarely seen in traditional covalent inhibitors. Finally, we demonstrated prolonged p53-pathway activation and more effective induction of the p53-mediated cell death in comparison to a noncovalent inhibitor. This study highlights the potential of the NASA warhead as a versatile electrophile for the covalent inhibition of PPIs and opens new avenues for the rational design of potent covalent PPI inhibitors.

Original language	English
Pages (from-to)	4766-4774
Number of pages	9
Journal	Journal of the American Chemical Society
Volume	143
Issue number	12
Early online date	18 Mar 2021
DOIs	https://doi.org/10.1021/jacs.1c00703
Publication status	Published - 31 Mar 2021

Bibliographical note

Funding Information:
We thank Eriko Kusaka (Kyoto University) for experimental support of the NMR measurements. We also thank Dr. Muneo Tsujikawa for plasmid construction. This work was funded by a Research Fellowship from the Japan Society for the Promotion of Science (JSPS) for Young Scientists to T.U., a Grant-in-Aid for Young Scientists (18K14334), and a Grant-in-Aid for Scientific Research on Innovative Areas “Integrated Bio-metal Science” (19H05764) to T.T., and Japan Science and Technology Agency (JST) ERATO grant JPMJER1802 to I.H. This work was also supported by a Grant-in-Aid for Scientific Research on Innovative Areas “Chemistry for Multimolecular Crowding Biosystems” (17H06348), EPSRC (EP/N013573/1). A.J.W. holds a Royal Society Leverhulme Trust Senior Fellowship (SRF/R1/191087). The MS raw data and analysis files have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the jPOST partner repository (http://jpostdb.org) with data set identifier PXD018105.

Publisher Copyright:
© 2021 American Chemical Society

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1021/jacs.1c00703

Cite this

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title = "Enhanced Suppression of a Protein-Protein Interaction in Cells Using Small-Molecule Covalent Inhibitors Based on an N-Acyl- N-alkyl Sulfonamide Warhead",

abstract = "Protein-protein interactions (PPIs) intimately govern various biological processes and disease states and therefore have been identified as attractive therapeutic targets for small-molecule drug discovery. However, the development of highly potent inhibitors for PPIs has proven to be extremely challenging with limited clinical success stories. Herein, we report irreversible inhibitors of the human double minute 2 (HDM2)/p53 PPI, which employ a reactive N-acyl-N-alkyl sulfonamide (NASA) group as a warhead. Mass-based analysis successfully revealed the kinetics of covalent inhibition and the modification sites on HDM2 to be the N-terminal α-amine and Tyr67, both rarely seen in traditional covalent inhibitors. Finally, we demonstrated prolonged p53-pathway activation and more effective induction of the p53-mediated cell death in comparison to a noncovalent inhibitor. This study highlights the potential of the NASA warhead as a versatile electrophile for the covalent inhibition of PPIs and opens new avenues for the rational design of potent covalent PPI inhibitors.",

author = "Tsuyoshi Ueda and Tomonori Tamura and Masaharu Kawano and Keiya Shiono and Fruzsina Hobor and Wilson, {Andrew J.} and Itaru Hamachi",

note = "Funding Information: We thank Eriko Kusaka (Kyoto University) for experimental support of the NMR measurements. We also thank Dr. Muneo Tsujikawa for plasmid construction. This work was funded by a Research Fellowship from the Japan Society for the Promotion of Science (JSPS) for Young Scientists to T.U., a Grant-in-Aid for Young Scientists (18K14334), and a Grant-in-Aid for Scientific Research on Innovative Areas “Integrated Bio-metal Science” (19H05764) to T.T., and Japan Science and Technology Agency (JST) ERATO grant JPMJER1802 to I.H. This work was also supported by a Grant-in-Aid for Scientific Research on Innovative Areas “Chemistry for Multimolecular Crowding Biosystems” (17H06348), EPSRC (EP/N013573/1). A.J.W. holds a Royal Society Leverhulme Trust Senior Fellowship (SRF/R1/191087). The MS raw data and analysis files have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the jPOST partner repository (http://jpostdb.org) with data set identifier PXD018105. Publisher Copyright: {\textcopyright} 2021 American Chemical Society",

year = "2021",

month = mar,

day = "31",

doi = "10.1021/jacs.1c00703",

language = "English",

volume = "143",

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Enhanced Suppression of a Protein-Protein Interaction in Cells Using Small-Molecule Covalent Inhibitors Based on an N-Acyl- N-alkyl Sulfonamide Warhead. / Ueda, Tsuyoshi; Tamura, Tomonori; Kawano, Masaharu et al.
In: Journal of the American Chemical Society, Vol. 143, No. 12, 31.03.2021, p. 4766-4774.

Research output: Contribution to journal › Article › peer-review

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AU - Ueda, Tsuyoshi

AU - Tamura, Tomonori

AU - Kawano, Masaharu

AU - Shiono, Keiya

AU - Hobor, Fruzsina

AU - Wilson, Andrew J.

AU - Hamachi, Itaru

N1 - Funding Information: We thank Eriko Kusaka (Kyoto University) for experimental support of the NMR measurements. We also thank Dr. Muneo Tsujikawa for plasmid construction. This work was funded by a Research Fellowship from the Japan Society for the Promotion of Science (JSPS) for Young Scientists to T.U., a Grant-in-Aid for Young Scientists (18K14334), and a Grant-in-Aid for Scientific Research on Innovative Areas “Integrated Bio-metal Science” (19H05764) to T.T., and Japan Science and Technology Agency (JST) ERATO grant JPMJER1802 to I.H. This work was also supported by a Grant-in-Aid for Scientific Research on Innovative Areas “Chemistry for Multimolecular Crowding Biosystems” (17H06348), EPSRC (EP/N013573/1). A.J.W. holds a Royal Society Leverhulme Trust Senior Fellowship (SRF/R1/191087). The MS raw data and analysis files have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the jPOST partner repository (http://jpostdb.org) with data set identifier PXD018105. Publisher Copyright: © 2021 American Chemical Society

PY - 2021/3/31

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N2 - Protein-protein interactions (PPIs) intimately govern various biological processes and disease states and therefore have been identified as attractive therapeutic targets for small-molecule drug discovery. However, the development of highly potent inhibitors for PPIs has proven to be extremely challenging with limited clinical success stories. Herein, we report irreversible inhibitors of the human double minute 2 (HDM2)/p53 PPI, which employ a reactive N-acyl-N-alkyl sulfonamide (NASA) group as a warhead. Mass-based analysis successfully revealed the kinetics of covalent inhibition and the modification sites on HDM2 to be the N-terminal α-amine and Tyr67, both rarely seen in traditional covalent inhibitors. Finally, we demonstrated prolonged p53-pathway activation and more effective induction of the p53-mediated cell death in comparison to a noncovalent inhibitor. This study highlights the potential of the NASA warhead as a versatile electrophile for the covalent inhibition of PPIs and opens new avenues for the rational design of potent covalent PPI inhibitors.

AB - Protein-protein interactions (PPIs) intimately govern various biological processes and disease states and therefore have been identified as attractive therapeutic targets for small-molecule drug discovery. However, the development of highly potent inhibitors for PPIs has proven to be extremely challenging with limited clinical success stories. Herein, we report irreversible inhibitors of the human double minute 2 (HDM2)/p53 PPI, which employ a reactive N-acyl-N-alkyl sulfonamide (NASA) group as a warhead. Mass-based analysis successfully revealed the kinetics of covalent inhibition and the modification sites on HDM2 to be the N-terminal α-amine and Tyr67, both rarely seen in traditional covalent inhibitors. Finally, we demonstrated prolonged p53-pathway activation and more effective induction of the p53-mediated cell death in comparison to a noncovalent inhibitor. This study highlights the potential of the NASA warhead as a versatile electrophile for the covalent inhibition of PPIs and opens new avenues for the rational design of potent covalent PPI inhibitors.

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DO - 10.1021/jacs.1c00703

M3 - Article

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SN - 0002-7863

VL - 143

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EP - 4774

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 12

ER -

Enhanced Suppression of a Protein-Protein Interaction in Cells Using Small-Molecule Covalent Inhibitors Based on an N-Acyl- N-alkyl Sulfonamide Warhead

Abstract

Bibliographical note

ASJC Scopus subject areas

UN SDGs

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