Abstract

The ubiquitin–proteasome system (UPS) comprises a network of enzymes that is responsible for maintaining cellular protein homeostasis. The therapeutic potential of this pathway has been validated by the clinical successes of a number of UPS modulators, including proteasome inhibitors and immunomodulatory imide drugs (IMiDs). Here we identified TAK-243 (formerly known as MLN7243) as a potent, mechanism-based small-molecule inhibitor of the ubiquitin activating enzyme (UAE), the primary mammalian E1 enzyme that regulates the ubiquitin conjugation cascade. TAK-243 treatment caused depletion of cellular ubiquitin conjugates, resulting in disruption of signaling events, induction of proteotoxic stress, and impairment of cell cycle progression and DNA damage repair pathways. TAK-243 treatment caused death of cancer cells and, in primary human xenograft studies, demonstrated antitumor activity at tolerated doses. Due to its specificity and potency, TAK-243 allows for interrogation of ubiquitin biology and for assessment of UAE inhibition as a new approach for cancer treatment.

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Acknowledgements

The authors would like to thank W. Harper (Harvard Medical School) for the UBA6-knockout and control MEFs, A. Berger for critical review of the manuscript, and E. Koenig and P. Shah for genomic data analysis. We would also like to thank J. Afroze, I. Bharathan, J. Gaulin, M. Girad, C. McIntyre, F. Soucy, T.T. Wong and Y. Ye for performing the chemical synthesis of TAK-243. All activities were completed and funded through Takeda Pharmaceuticals Inc.

Author information

Author notes

    • Marc L Hyer
    •  & Neil F Bence

    Present addresses: Agios Pharmaceuticals, Cambridge, Massachusetts, USA (M.L.H.) and Nurix, Inc., San Francisco, California, USA (N.F.B.).

    • Marc L Hyer
    •  & Michael A Milhollen

    These authors contributed equally to this work.

Affiliations

  1. Takeda Pharmaceuticals Inc., Cambridge, Massachusetts, USA.

    • Marc L Hyer
    • , Michael A Milhollen
    • , Jeff Ciavarri
    • , Paul Fleming
    • , Tary Traore
    • , Darshan Sappal
    • , Jessica Huck
    • , Judy Shi
    • , James Gavin
    • , Jim Brownell
    • , Yu Yang
    • , Bradley Stringer
    • , Robert Griffin
    • , Frank Bruzzese
    • , Teresa Soucy
    • , Jennifer Duffy
    • , Claudia Rabino
    • , Jessica Riceberg
    • , Kara Hoar
    • , Anya Lublinsky
    • , Saurabh Menon
    • , Michael Sintchak
    • , Nancy Bump
    • , Sai M Pulukuri
    • , Steve Langston
    • , Stephen Tirrell
    • , Mike Kuranda
    • , Petter Veiby
    • , John Newcomb
    • , Ping Li
    • , Jing Tao Wu
    • , Josh Powe
    • , Lawrence R Dick
    • , Paul Greenspan
    • , Katherine Galvin
    • , Mark Manfredi
    • , Chris Claiborne
    • , Benjamin S Amidon
    •  & Neil F Bence

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Contributions

M.L.H., M.A.M., N.F.B., B.S.A., J.G. and L.R.D. participated in writing, reviewing and editing of the manuscript; M.L.H., M.A.M. and N.F.B. participated in the planning, initiation, data generation and analysis of biological experiments; J.C., S.L. and P.F. participated in the planning, initiation, design and execution of chemical synthesis; M.S. and N.B. performed crystallography experiments; J.P. was the toxicology representative on the program; J.G., T.S., F.B. and J.B. performed biochemical analyses; M.A.M., D.S., J.D., C.R., J.R. and K.H. performed in vitro cell culture experiments; M.L.H., T.T., J.H., J.S. and S.M.P. performed in vivo antitumor activity and pharmacodynamic experiments; A.L. and S.M. evaluated compound potencies in cell-based assays; Y.Y. and B.S. performed immunohistochemistry experiments; R.G. performed pharmacokinetic analyses; B.S.A., S.T., M.K., P.V., J.N., P.L., J.T.W., P.G., K.G., M.M. and C.C. provided project oversight and review.

Competing interests

All of the authors were employees of Takeda Pharmaceuticals at the time of these studies.

Corresponding author

Correspondence to Michael A Milhollen.

Supplementary information

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    Supplementary Figures & Tables

    Supplementary Figures 1–16 & Supplementary Tables 1, 3

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    Life Sciences Reporting Summary

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    Supplementary Table 2

    Selectivity profiling of TAK-243 against kinases, cellular receptors (Novascreen) and carbonic anhydrases.

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DOI

https://doi.org/10.1038/nm.4474