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An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer

Abstract

The clinical development of an inhibitor of cellular proteasome function suggests that compounds targeting other components of the ubiquitin–proteasome system might prove useful for the treatment of human malignancies. NEDD8-activating enzyme (NAE) is an essential component of the NEDD8 conjugation pathway that controls the activity of the cullin-RING subtype of ubiquitin ligases, thereby regulating the turnover of a subset of proteins upstream of the proteasome. Substrates of cullin-RING ligases have important roles in cellular processes associated with cancer cell growth and survival pathways. Here we describe MLN4924, a potent and selective inhibitor of NAE. MLN4924 disrupts cullin-RING ligase-mediated protein turnover leading to apoptotic death in human tumour cells by a new mechanism of action, the deregulation of S-phase DNA synthesis. MLN4924 suppressed the growth of human tumour xenografts in mice at compound exposures that were well tolerated. Our data suggest that NAE inhibitors may hold promise for the treatment of cancer.

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Figure 1: MLN4924 is a potent and selective inhibitor of NEDD8-activating enzyme.
Figure 2: NEDD8 pathway inhibition, in contrast to proteasome inhibition, minimally affects bulk intracellular protein turnover.
Figure 3: Inhibition of the NEDD8 pathway in cells results in S-phase defects, DNA damage and apoptosis.
Figure 4: MLN4924 inhibits the NEDD8 pathway resulting in DNA damage in human tumour xenografts.
Figure 5: MLN4924 produces tumour growth inhibition in human tumour xenografts.

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Acknowledgements

As this project progressed through the discovery phase, many scientists made significant contributions to its success. The authors would like to thank all past contributors to the NAE project including M. H. Foley, V. Chau, T. Gladysheva and S. Sadis. We would also like to thank D. Bowman for technical help with imaging assays, J. Blank for technical help with protein turnover assays and A. Burkhardt, P. Veiby, M. Manfredi and B. Hibner for support and guidance.

Author Contributions T.A.S., P.G.S., M.A.M., A.J.B. and J.M.G. participated in the planning, initiation, data generation and analysis of biological experiments and preparation of the manuscript. C.A.C., E.J.O., S.V., G.S.W., S.C., S.P. and S.P.L. participated in the planning, initiation, design and execution of chemical synthesis. S.A., D.P.C., A.D., H.M., M.R., J.L.G., R.E.G. and T.Talreja performed chemical synthesis; U.N. performed in vitro cell culture experiments; T.Traore and J.Z. performed in vivo anti-tumour activity and pharmacodynamic experiments; J.Y., J.J.G. and M.P.T. performed pharmacodynamic analysis experiments; A.R.L. evaluated compound potencies in cell-based assays; K.E.B. and A.M. performed immunohistochemical experiments; M.D.S. performed crystallography studies. L.R.D., J.E.B., C.F.C., M.R. and J.B.B. provided oversight of the project, reviewed and edited the manuscript.

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Correspondence to Teresa A. Soucy.

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[Competing Interests: All authors were employees and stockholders of Millennium Pharmaceuticals Inc. during the time the studies were completed.]

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MLN4924 will be provided once a standard Materials Transfer Agreement has been executed.

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This file contains Supplementary Figures 1-6 with Legends, Supplementary Tables 1-2, Supplementary Methods and a Supplementary Discussion and Supplementary References. (PDF 523 kb)

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Soucy, T., Smith, P., Milhollen, M. et al. An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer. Nature 458, 732–736 (2009). https://doi.org/10.1038/nature07884

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