Abstract

Avoidance of apoptosis is critical for the development and sustained growth of tumours. The pro-survival protein myeloid cell leukemia 1 (MCL1) is overexpressed in many cancers, but the development of small molecules targeting this protein that are amenable for clinical testing has been challenging. Here we describe S63845, a small molecule that specifically binds with high affinity to the BH3-binding groove of MCL1. Our mechanistic studies demonstrate that S63845 potently kills MCL1-dependent cancer cells, including multiple myeloma, leukaemia and lymphoma cells, by activating the BAX/BAK-dependent mitochondrial apoptotic pathway. In vivo, S63845 shows potent anti-tumour activity with an acceptable safety margin as a single agent in several cancers. Moreover, MCL1 inhibition, either alone or in combination with other anti-cancer drugs, proved effective against several solid cancer-derived cell lines. These results point towards MCL1 as a target for the treatment of a wide range of tumours.

  • Subscribe to Nature for full access:

    $199

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    & Cell death signaling. Cold Spring Harb. Perspect. Biol. 7, 1–24 (2015)

  2. 2.

    & Hallmarks of cancer: the next generation. Cell 144, 646–674 (2011)

  3. 3.

    , , & Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nat. Rev. Mol. Cell Biol. 15, 49–63 (2014)

  4. 4.

    et al. Rationale for Bcl-xL/Bad peptide complex formation from structure, mutagenesis, and biophysical studies. Protein Sci. 9, 2528–2534 (2000)

  5. 5.

    , , & Thirty years of BCL-2: translating cell death discoveries into novel cancer therapies. Nat. Rev. Cancer 16, 99–109 (2016)

  6. 6.

    et al. Navitoclax, a targeted high-affinity inhibitor of BCL-2, in lymphoid malignancies: a phase 1 dose-escalation study of safety, pharmacokinetics, pharmacodynamics, and antitumour activity. Lancet Oncol. 11, 1149–1159 (2010)

  7. 7.

    et al. Substantial susceptibility of chronic lymphocytic leukemia to BCL2 inhibition: results of a phase I study of navitoclax in patients with relapsed or refractory disease. J. Clin. Oncol. 30, 488–496 (2012)

  8. 8.

    et al. Targeting BCL2 with venetoclax in relapsed chronic lymphocytic leukemia. N. Engl. J. Med. 374, 311–322 (2016)

  9. 9.

    , , , & Decoding and unlocking the BCL-2 dependency of cancer cells. Nat. Rev. Cancer 13, 455–465 (2013)

  10. 10.

    et al. The landscape of somatic copy-number alteration across human cancers. Nature 463, 899–905 (2010)

  11. 11.

    Mcl-1 is a potential therapeutic target in multiple types of cancer. Cell. Mol. Life Sci. 66, 1326–1336 (2009)

  12. 12.

    et al. Anti-apoptotic Mcl-1 is essential for the development and sustained growth of acute myeloid leukemia. Genes Dev. 26, 120–125 (2012)

  13. 13.

    et al. Targeting of MCL-1 kills MYC-driven mouse and human lymphomas even when they bear mutations in p53. Genes Dev. 28, 58–70 (2014)

  14. 14.

    et al. Requirement for antiapoptotic MCL-1 in the survival of BCR–ABL B-lineage acute lymphoblastic leukemia. Blood 122, 1587–1598 (2013)

  15. 15.

    , , & MCL-1 but not BCL-XL is critical for the development and sustained expansion of thymic lymphoma in p53-deficient mice. Blood 124, 3939–3946 (2014)

  16. 16.

    et al. Re-activation of mitochondrial apoptosis inhibits T-cell lymphoma survival and treatment resistance. Leukemia 30, 1520–1530 (2016)

  17. 17.

    et al. Hierarchy for targeting pro-survival BCL2 family proteins in multiple myeloma: pivotal role of MCL1. Blood blood-2016- 03–704908 (2016)

  18. 18.

    et al. Chemical genomics identifies small-molecule MCL1 repressors and BCL-XL as a predictor of MCL1 dependency. Cancer Cell 21, 547–562 (2012)

  19. 19.

    et al. Potent and selective small-molecule MCL-1 inhibitors demonstrate on-target cancer cell killing activity as single agents and in combination with ABT-263 (navitoclax). Cell Death Dis. 6, e1590 (2015)

  20. 20.

    et al. Discovery of 2-indole-acylsulfonamide myeloid cell leukemia 1 (Mcl-1) inhibitors using fragment-based methods. J. Med. Chem. 59, 2054–2066 (2016)

  21. 21.

    , , & Small molecules targeting Mcl-1: the search for a silver bullet in cancer therapy. MedChemComm 7, 778–787 (2016)

  22. 22.

    et al. Obligate role of anti-apoptotic MCL-1 in the survival of hematopoietic stem cells. Science 307, 1101–1104 (2005)

  23. 23.

    et al. Deletion of MCL-1 causes lethal cardiac failure and mitochondrial dysfunction. Genes Dev. 27, 1351–1364 (2013)

  24. 24.

    et al. Loss of MCL-1 leads to impaired autophagy and rapid development of heart failure. Genes Dev. 27, 1365–1377 (2013)

  25. 25.

    , & BCL-2 family antagonists for cancer therapy. Nat. Rev. Drug Discov. 7, 989–1000 (2008)

  26. 26.

    et al. Structure-guided design of a series of MCL-1 inhibitors with high affinity and selectivity. J. Med. Chem. 58, 2180–2194 (2015)

  27. 27.

    , & Mcl-1 stability determines mitotic cell fate of human multiple myeloma tumor cells treated with the kinesin spindle protein inhibitor ARRY-520. Mol. Cancer Ther. 9, 2046–2056 (2010)

  28. 28.

    et al. An inhibitor of Bcl-2 family proteins induces regression of solid tumours. Nature 435, 677–681 (2005)

  29. 29.

    et al. ABT-199, a potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets. Nat. Med. 19, 202–208 (2013)

  30. 30.

    et al. An inducible lentiviral guide RNA platform enables the identification of tumor-essential genes and tumor-promoting mutations in vivo. Cell Reports 10, 1422–1432 (2015)

  31. 31.

    et al. Antisense strategy shows that Mcl-1 rather than Bcl-2 or Bcl-x(L) is an essential survival protein of human myeloma cells. Blood 100, 194–199 (2002)

  32. 32.

    , & Myeloid cell factor-1 is a critical survival factor for multiple myeloma. Blood 99, 1885–1893 (2002)

  33. 33.

    et al. A novel BH3 ligand that selectively targets Mcl-1 reveals that apoptosis can proceed without Mcl-1 degradation. J. Cell Biol. 180, 341–355 (2008)

  34. 34.

    et al. Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol. Cell 17, 393–403 (2005)

  35. 35.

    et al. The Bcl-2 specific BH3 mimetic ABT-199: a promising targeted therapy for t(11;14) multiple myeloma. Leukemia 28, 210–212 (2014)

  36. 36.

    et al. MB4-2 breakpoint in MMSET combined with del(17p) defines a subset of t(4;14) multiple myeloma with very poor prognosis. Haematologica 100, e471–e474 (2015)

  37. 37.

    et al. Mcl-1 is critical for survival in a subgroup of non-small-cell lung cancer cell lines. Oncogene 30, 1963–1968 (2011)

  38. 38.

    et al. MCL-1 Is a key determinant of breast cancer cell survival: validation of MCL-1 dependency utilizing a highly selective small molecule inhibitor. Mol. Cancer Ther. 14, 1837–1847 (2015)

  39. 39.

    , , & Unleashing the power of inhibitors of oncogenic kinases through BH3 mimetics. Nat. Rev. Cancer 9, 321–326 (2009)

  40. 40.

    et al. Treatment of B-RAF mutant human tumor cells with a MEK inhibitor requires Bim and is enhanced by a BH3 mimetic. J. Clin. Invest. 118, 3651–3659 (2008)

  41. 41.

    et al. Synthetic lethal interaction of combined BCL-XL and MEK inhibition promotes tumor regressions in KRAS mutant cancer models. Cancer Cell 23, 121–128 (2013)

  42. 42.

    et al. Anti-apoptotic MCL-1 localizes to the mitochondrial matrix and couples mitochondrial fusion to respiration. Nat. Cell Biol. 14, 575–583 (2012)

  43. 43.

    et al. Single diastereomer of a macrolactam core binds specifically to myeloid cell leukemia 1 (MCL1). ACS Med. Chem. Lett. 5, 1308–1312 (2014)

  44. 44.

    , , & A synergistic approach to protein crystallization: combination of a fixed-arm carrier with surface entropy reduction. Protein Sci. 19, 901–913 (2010)

  45. 45.

    XDS. Acta Crystallogr. D Biol. Crystallogr. 66, 125–132 (2010)

  46. 46.

    & MOLREP: an automated program for molecular replacement. J. Appl. Cryst. 30, 1022–1025 (1997)

  47. 47.

    , & Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D Biol. Crystallogr. 53, 240–255 (1997)

  48. 48.

    et al. Development and optimization of a binding assay for the XIAP BIR3 domain using fluorescence polarization. Anal. Biochem. 332, 261–273 (2004)

  49. 49.

    , , & Off-rate screening (ORS) by surface plasmon resonance. An efficient method to kinetically sample hit to lead chemical space from unpurified reaction products. J. Med. Chem. 57, 2845–2850 (2014)

  50. 50.

    et al. Fragment screening by weak affinity chromatography: comparison with established techniques for screening against HSP90. Anal. Chem. 85, 6756–6766 (2013)

  51. 51.

    et al. The c-myc oncogene driven by immunoglobulin enhancers induces lymphoid malignancy in transgenic mice. Nature 318, 533–538 (1985)

Download references

Acknowledgements

We thank S. Courtade-Gaiani and D. Valour for bioinformatics support, E. Borges for assistance on manuscript formatting, E. Schneider and C. Wagner-Legrand, H. Johnson, G. Siciliano and K. Hughes for technical help for in vivo studies, N. Whitehead for protein production support, P. Bouillet and L. A. O’Reilly for assistance with histology, M. Fallowfield, J. D’Alessandro, L. Terry, V. Lemesre, J.-P. Galizzi and C. de la Moureyre for in vitro assay support, H. Simmonite for analytical support and L. Andrieu, L. Montane and A. Schmutz for biostatistical support. Research at WEHI is supported by the National Health and Medical Research Council Australia (NHMRC, GNT1016647, GNT1016701, GNT1020363, GNT1086291, GNT1049720, GNT1057742, GNT1079560), the Leukemia and Lymphoma Society (SCOR grant 7001-03), The Cancer Council (1086157 GLK, grant in aid to A.W.R. and D.C.S.H.), The Kay Kendall Leukemia Fund Intermediate Fellowship (KKL331 to G.L.K.), the Victoria Cancer Agency, the Australian Cancer Research Foundation, a Victorian State Government Operational Infrastructure Support (OIS) grant and the estate of Anthony (Toni) Redstone OAM.

Author information

Affiliations

  1. Servier Research Institute of Medicinal Chemistry, Budapest 1031, Hungary

    • András Kotschy
    • , Zoltán Szlavik
    • , Márton Csekei
    • , Attila Paczal
    • , Zoltán B. Szabo
    • , Szabolcs Sipos
    • , Gábor Radics
    • , Agnes Proszenyak
    • , Balázs Balint
    • , Levente Ondi
    •  & Gábor Blasko
  2. Vernalis (R&D) Ltd., Cambridge CB21 6GB, UK

    • James Murray
    • , James Davidson
    • , Alan Robertson
    • , Allan Surgenor
    • , Pawel Dokurno
    • , Ijen Chen
    • , Natalia Matassova
    • , Julia Smith
    • , Christopher Pedder
    •  & Christopher Graham
  3. Institut de Recherches Servier Oncology R&D Unit, Croissy Sur Seine 78290, France

    • Ana Leticia Maragno
    • , Gaëtane Le Toumelin-Braizat
    • , Maïa Chanrion
    • , Alain Bruno
    • , Aurélie Studeny
    • , Gaëlle Lysiak-Auvity
    • , Anne-Marie Girard
    • , Fabienne Gravé
    • , Ghislaine Guasconi
    • , Nicolas Cauquil
    • , Frédéric Colland
    • , John A. Hickman
    •  & Olivier Geneste
  4. The Walter and Eliza Hall Institute of Medical Research, Melbourne 3052, Australia

    • Gemma L. Kelly
    • , Jia-Nan Gong
    • , David Segal
    • , Chris D. Riffkin
    • , Laura C. A. Galbraith
    • , Brandon J. Aubrey
    • , Margs S. Brennan
    • , Marco J. Herold
    • , Catherine Chang
    • , Andrew W. Roberts
    • , David C. S. Huang
    • , Andreas Strasser
    •  & Guillaume Lessene
  5. Department of Medical Biology, University of Melbourne, Melbourne 3010, Australia

    • Gemma L. Kelly
    • , Jia-Nan Gong
    • , David Segal
    • , Chris D. Riffkin
    • , Laura C. A. Galbraith
    • , Brandon J. Aubrey
    • , Margs S. Brennan
    • , Marco J. Herold
    • , Catherine Chang
    • , Andrew W. Roberts
    • , David C. S. Huang
    • , Andreas Strasser
    •  & Guillaume Lessene
  6. Australian Centre for Blood Diseases, Monash University, Melbourne 3004, Australia

    • Donia M. Moujalled
    • , Giovanna Pomilio
    •  & Andrew H. Wei
  7. Department of Clinical Haematology and Bone Marrow Transplantation, The Royal Melbourne Hospital, Victorian Comprehensive Cancer Centre, Melbourne 3050, Australia

    • Brandon J. Aubrey
    •  & Andrew W. Roberts
  8. Institut de Recherches Servier, Biomarker Research Division, Croissy Sur Seine 78290, France.

    • Fabien Melchiore
    • , Nolwen Guigal-Stephan
    •  & Brian Lockhart
  9. Faculty of Medicine, The University of Melbourne, Melbourne 3010, Australia.

    • Andrew W. Roberts
  10. Department of Clinical Haematology, The Alfred Hospital, Melbourne 3004, Australia.

    • Andrew H. Wei
  11. Department of Pharmacology and Pharmaceutics, The University of Melbourne, Melbourne 3010, Australia

    • Guillaume Lessene

Authors

  1. Search for András Kotschy in:

  2. Search for Zoltán Szlavik in:

  3. Search for James Murray in:

  4. Search for James Davidson in:

  5. Search for Ana Leticia Maragno in:

  6. Search for Gaëtane Le Toumelin-Braizat in:

  7. Search for Maïa Chanrion in:

  8. Search for Gemma L. Kelly in:

  9. Search for Jia-Nan Gong in:

  10. Search for Donia M. Moujalled in:

  11. Search for Alain Bruno in:

  12. Search for Márton Csekei in:

  13. Search for Attila Paczal in:

  14. Search for Zoltán B. Szabo in:

  15. Search for Szabolcs Sipos in:

  16. Search for Gábor Radics in:

  17. Search for Agnes Proszenyak in:

  18. Search for Balázs Balint in:

  19. Search for Levente Ondi in:

  20. Search for Gábor Blasko in:

  21. Search for Alan Robertson in:

  22. Search for Allan Surgenor in:

  23. Search for Pawel Dokurno in:

  24. Search for Ijen Chen in:

  25. Search for Natalia Matassova in:

  26. Search for Julia Smith in:

  27. Search for Christopher Pedder in:

  28. Search for Christopher Graham in:

  29. Search for Aurélie Studeny in:

  30. Search for Gaëlle Lysiak-Auvity in:

  31. Search for Anne-Marie Girard in:

  32. Search for Fabienne Gravé in:

  33. Search for David Segal in:

  34. Search for Chris D. Riffkin in:

  35. Search for Giovanna Pomilio in:

  36. Search for Laura C. A. Galbraith in:

  37. Search for Brandon J. Aubrey in:

  38. Search for Margs S. Brennan in:

  39. Search for Marco J. Herold in:

  40. Search for Catherine Chang in:

  41. Search for Ghislaine Guasconi in:

  42. Search for Nicolas Cauquil in:

  43. Search for Fabien Melchiore in:

  44. Search for Nolwen Guigal-Stephan in:

  45. Search for Brian Lockhart in:

  46. Search for Frédéric Colland in:

  47. Search for John A. Hickman in:

  48. Search for Andrew W. Roberts in:

  49. Search for David C. S. Huang in:

  50. Search for Andrew H. Wei in:

  51. Search for Andreas Strasser in:

  52. Search for Guillaume Lessene in:

  53. Search for Olivier Geneste in:

Contributions

A.K., Z.S., J.D., M.Cs., A.Pa., Z.B.S., S.S., G.R., A.Pr., B.B., L.O., G.B. and C.G. supervised and performed the chemistry. A.K., Z.B.S., J.M., J.D. and I.C. performed the drug design and molecular modelling. A.L.M., G.L.T.-B., G.L.K., J.-N.G., D.M.M., A.Stu., D.S., C.D.R., G.P., C.C., G.G. and N.C. performed cell based experiments. G.L.K., G.L.-A., A.-M.G., F.G., M.S.B., L.C.A.G. and M.J.H. performed the in vivo experiments. B.J.A. contributed to the histology analysis. J.M., A.R., A.Su., P.D., N.M., J.S. and C.P. produced recombinant proteins, performed biochemical assays and crystallographic studies. M.Ch., G.L.K., A.B. and M.J.H. designed the in vivo experiments. F.M., N.G.-S. and B.L. designed and performed the bioinformatic analysis. A.-L.M., J.M., J.D., G.L.K., F.C., J.A.H., A.W.R., D.C.S.H., A.H.W., A.Str., G.L. and O.G. supervised the studies, designed the experiments and interpreted the results. A.Str., G.L., D.C.S.H. and O.G. wrote the manuscript with the assistance of A.L.M., G.L.K. and the other authors.

Competing interests

G.L.T.-B., A.L.M., G.G., A.Stu., N.C., M.Ch., A.B., G.L.-A., A.-M.G., F.G., O.G., J.A.H., B.L., N.G.-S., F.C., F.M., A.K., Z.Szl., M.Cs., A.Pa., Z.Sza., S.S., G.R., L.O., A.Pr., B.B. and G.B. are employees of Servier. N.M., C.P., J.S., I.C., C.G., J.D., A.R., A.Su., P.D. and J.M. are employees of Vernalis (R&D) Ltd., G.L., A.Str., G.L.K., M.J.H., J.G., D.S., C.D.R., A.W.R., C.C., M.S.B. and D.C.S.H. are employees of the Walter and Eliza Hall Institute of Medical Research, which receives research funding and milestone payments in relation to venetoclax (ABT-199). G.L., A.Str., G.L.K., M.J.H., J.-N.G., D.S., C.D.R., A.W.R., M.S.B. and D.C.S.H. receive research funding from Servier. A.H.W. serves on the advisory board for Servier and receives research funding from Servier.

Corresponding author

Correspondence to Olivier Geneste.

Reviewer Information Nature thanks S. Fletcher, F. Stegmeier, G. Wagner and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Extended data

Extended data figures

  1. 1.

    Biophysical characterization of the binding of MCL1 inhibitors to human and mouse MCL1 and serum effect on their cellular potency.

  2. 2.

    Targeting MCL1 genetically in H929 cells induces caspase-mediated cell death.

  3. 3.

    Impact of treatment with S63845 on the interaction of MCL1 with pro-apoptotic BCL-2 family members and on the level and stability of MCL1.

  4. 4.

    S63845 induces apoptosis of sensitive tumour derived cell lines.

  5. 5.

    Correlation between the sensitivity of multiple-myeloma-derived cell lines to S63845 and killing by the MCL1-selective ligand BIM2A. BAX/BAK dependency for S63845-induced killing in KMS-12-PE and AMO1 myeloma cells. Expression of pro-survival BCL-2 family proteins across a panel of multiple-myeloma-derived lines.

  6. 6.

    Anti-tumour efficacy and effect of dose scheduling of S63845 in H929 and AMO1 multiple myeloma xenograft models. The in vitro activity of S63845 in human Burkitt-lymphoma-derived cell lines and in vivo activity of S63845 on individual Eμ-Myc lymphoma cell lines.

  7. 7.

    Spearman correlations distribution in haematological cell lines and AML data on patient samples.

  8. 8.

    S63845 activity in solid tumour-derived cell lines, correlation with BCL-XL mRNA expression and impact of treatment with the MEK1/2 inhibitor, trametenib, the HER-2 inhibitor, lapatinib, the B-RAF inhibitor, PLX-4032, or the EGFR inhibitor, tarceva, on the levels of p-ERK and the pro-apoptotic protein BIM.

  9. 9.

    Tolerability of S63845 in mice and impact of treatment on healthy tissues at doses that prevent tumour expansion.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-2, Supplementary Tables 1-2 and Supplementary Methods.

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.