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Organometallic palladium reagents for cysteine bioconjugation

Nature volume 526, pages 687–691 (2015)Cite this article

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Abstract

Reactions based on transition metals have found wide use in organic synthesis, in particular for the functionalization of small molecules1,2. However, there are very few reports of using transition-metal-based reactions to modify complex biomolecules3,4, which is due to the need for stringent reaction conditions (for example, aqueous media, low temperature and mild pH) and the existence of multiple reactive functional groups found in biomolecules. Here we report that palladium(ii) complexes can be used for efficient and highly selective cysteine conjugation (bioconjugation) reactions that are rapid and robust under a range of bio-compatible reaction conditions. The straightforward synthesis of the palladium reagents from diverse and easily accessible aryl halide and trifluoromethanesulfonate precursors makes the method highly practical, providing access to a large structural space for protein modification. The resulting aryl bioconjugates are stable towards acids, bases, oxidants and external thiol nucleophiles. The broad utility of the bioconjugation platform was further corroborated by the synthesis of new classes of stapled peptides and antibody–drug conjugates. These palladium complexes show potential as benchtop reagents for diverse bioconjugation applications.

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Figure 1: Organometallic palladium reagents for cysteine modification.
Figure 2: The substrate scope of cysteine arylation using organometallic palladium reagents.
Figure 3: Protein modification using the developed palladium reagents.
Figure 4: Peptide stapling and antibody drug conjugate formation using palladium-based reagents.

References

  1. Crabtree, R. H. in The Organometallic Chemistry of the Transition Metals 6th edn (Wiley, 2014)

    Book  Google Scholar 

  2. Diederich, F. & Stang, P. J. (eds) Metal-catalyzed Cross-coupling Reactions (Wiley & Sons, 2008)

    Google Scholar 

  3. Antos, J. M. & Francis, M. B. Transition metal catalyzed methods for site-selective protein modification. Curr. Opin. Chem. Biol. 10, 253–262 (2006)

    Article  CAS  Google Scholar 

  4. Yang, M., Li, J. & Chen, P. R. Transition metal-mediated bioorthogonal protein chemistry in living cells. Chem. Soc. Rev. 43, 6511–6526 (2014)

    Article  CAS  Google Scholar 

  5. Walsh, C. T., Garneau-Tsodikova, S. & Gatto, G. J. Protein posttranslational modifications: the chemistry of proteome diversifications. Angew. Chem. Int. Edn 44, 7342–7372 (2005)

    Article  CAS  Google Scholar 

  6. Rabuka, D. Chemoenzymatic methods for site-specific protein modification. Curr. Opin. Chem. Biol. 14, 790–796 (2010)

    Article  CAS  Google Scholar 

  7. Spicer, C. D. & Davis, B. G. Selective chemical protein modification. Nature Commun. 5, 4740 (2014)

    Article  CAS  ADS  Google Scholar 

  8. Chalker, J. M., Bernardes, G. J. L., Lin, Y. A. & Davis, B. G. Chemical modification of proteins at cysteine: opportunities in chemistry and biology. Chem. Asian J. 4, 630–640 (2009)

    Article  CAS  Google Scholar 

  9. Cal, P. M. S. D., Bernardes, G. J. L. & Gois, P. M. P. Cysteine-selective reactions for antibody conjugation. Angew. Chem. Int. Edn 53, 10585–10587 (2014)

    Article  CAS  Google Scholar 

  10. Toda, N., Asano, S. & Barbas, C. F. Rapid, stable, chemoselective labeling of thiols with Julia–Kocieński-like reagents: a serum-stable alternative to maleimide based protein conjugation. Angew. Chem. Int. Edn 52, 12592–12596 (2013)

    Article  CAS  Google Scholar 

  11. Lyon, R. P. et al. Self-hydrolyzing maleimides improve the stability and pharmacological properties of antibody-drug conjugates. Nature Biotechnol. 32, 1059–1062 (2014)

    Article  CAS  Google Scholar 

  12. Simmons, R. L., Yu, R. T. & Myers, A. G. Storable arylpalladium(II) reagents for alkene labeling in aqueous media. J. Am. Chem. Soc. 133, 15870–15873 (2011)

    Article  CAS  Google Scholar 

  13. Cheng, G., Lim, R. K. V., Li, N. & Lin, Q. Storable palladacycles for selective functionalization of alkyne-containing proteins. Chem. Commun. 49, 6809–6811 (2013)

    Article  CAS  Google Scholar 

  14. Barder, T. E., Biscoe, M. R. & Buchwald, S. L. Structural insights into active catalyst structures and oxidative addition to (biaryl)phosphine-palladium complexes via density functional theory and experimental studies. Organometallics 26, 2183–2192 (2007)

    Article  CAS  Google Scholar 

  15. Lee, C.-F., Liu, Y.-C. & Badsara, S. S. Transition-metal-catalyzed C–S bond coupling reaction. Chem. Asian J. 9, 706–722 (2014)

    Article  CAS  Google Scholar 

  16. Bong, D. T. & Ghadiri, M. R. Chemoselective Pd(0)-catalyzed peptide coupling in water. Org. Lett. 3, 2509–2511 (2001)

    Article  CAS  Google Scholar 

  17. Korneeva, E. N., Ovchinnikov, M. V. & Kostić, N. M. Peptide hydrolysis promoted by polynuclear and organometallic complexes of palladium(II) and platinum(II). Inorg. Chim. Acta 243, 9–13 (1996)

    Article  CAS  Google Scholar 

  18. Tilley, S. D. & Francis, M. B. Tyrosine-selective protein alkylation using π-allylpalladium complexes. J. Am. Chem. Soc. 128, 1080–1081 (2006)

    Article  CAS  Google Scholar 

  19. Shafer, D. E., Inman, J. K. & Lees, A. Reaction of tris(2-carboxyethyl)phosphine (TCEP) with maleimide and α-haloacyl groups: anomalous elution of TCEP by gel filtration. Anal. Biochem. 282, 161–164 (2000)

    Article  CAS  Google Scholar 

  20. Gorin, G., Martic, P. A. & Doughty, G. Kinetics of the reaction of N-ethylmaleimide with cysteine and some congeners. Arch. Biochem. Biophys. 115, 593–597 (1966)

    Article  CAS  Google Scholar 

  21. Gilbreth, R. N. & Koide, S. Structural insights for engineering binding proteins based on non-antibody scaffolds. Curr. Opin. Struct. Biol. 22, 413–420 (2012)

    Article  CAS  Google Scholar 

  22. Kung, K. K.-Y. et al. Cyclometalated gold(III) complexes for chemoselective cysteine modification via ligand controlled C–S bond-forming reductive elimination. Chem. Commun. 50, 11899–11902 (2014)

    Article  CAS  Google Scholar 

  23. Arora, N. & Leppla, S. H. Fusions of anthrax toxin lethal factor with shiga toxin and diphtheria toxin enzymatic domains are toxic to mammalian cells. Infect. Immun. 62, 4955–4961 (1994)

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Bird, G. H., Gavathiotis, E., LaBelle, J. L., Katz, S. G. & Walensky, L. D. Distinct BimBH3 (BimSAHB) stapled peptides for structural and cellular studies. ACS Chem. Biol. 9, 831–837 (2014)

    Article  CAS  Google Scholar 

  25. Verdine, G. L. & Hilinski, G. J. Stapled peptides for intracellular drug targets. Methods Enzymol. 503, 3–33 (2012)

    Article  CAS  Google Scholar 

  26. Lau, Y. H., de Andrade, P., Wu, Y. & Spring, D. R. Peptide stapling techniques based on different macrocyclization chemistries. Chem. Soc. Rev. 44, 91–102 (2015)

    Article  CAS  Google Scholar 

  27. Writing the macrocycle manual. Nature Chem. Biol. 10, 693 (2014)

  28. Spokoyny, A. M. et al. A perfluoroaryl-cysteine SNAr chemistry approach to unprotected peptide stapling. J. Am. Chem. Soc. 135, 5946–5949 (2013)

    Article  CAS  Google Scholar 

  29. Chari, R. V. J., Miller, M. L. & Widdison, W. C. Antibody-drug conjugates: an emerging concept in cancer therapy. Angew. Chem. Int. Edn 53, 3796–3827 (2014)

    Article  CAS  Google Scholar 

  30. Sun, M. M. C. et al. Reduction-alkylation strategies for the modification of specific monoclonal antibody disulfides. Bioconjug. Chem. 16, 1282–1290 (2005)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Financial support for this work was provided by the National Institutes of Health (GM-58160; GM-110535; postdoctoral fellowship for A.M.S., 1F32GM101762), the MIT start-up fund (B.L.P.), a Damon Runyon Cancer Research Foundation Award (B.L.P.) and the Sontag Foundation Distinguished Scientist Award (B.L.P.). C.Z. is the recipient of the George Büchi Research Fellowship and the Koch Graduate Fellowship in Cancer Research of MIT. We thank R. J. Collier (Harvard) for contributing select laboratory equipment used in this study. We thank the Biological Instrument Facility of MIT for providing the Octet BioLayer Interferometry System (NSF S10 OD016326). We are indebted to the NERCE facility (U54A1057159) for expressing the toxin proteins. We thank M. Lu and A. Rabideu for help with cell assays. The Varian 300 spectrometer used for portions of this work was purchased with funds from NSF (grant CHE-9808061). The departmental X-ray diffraction instrumentation was purchased with the help of funding from NSF (CHE-0946721). We are grateful to P. Müller (MIT) for X-ray crystallographic analysis of 1A–OTf˙CH3CN and to A. Rancier (Merck) for the ICP-MS analysis.

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Author notes

  1. Alexander M. Spokoyny

    Present address: †Present address: Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095, USA.,

  2. Ekaterina V. Vinogradova and Chi Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Ekaterina V. Vinogradova, Chi Zhang, Alexander M. Spokoyny, Bradley L. Pentelute & Stephen L. Buchwald

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  1. Ekaterina V. Vinogradova
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  2. Chi Zhang
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  3. Alexander M. Spokoyny
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  4. Bradley L. Pentelute
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Contributions

S.L.B. conceived the idea of using palladium(ii) reagents for bioconjugation; E.V.V., C.Z., A.M.S., B.L.P. and S.L.B. designed the research; E.V.V. and C.Z. conducted the majority of the experimental work; and A.M.S. conducted initial feasibility experiments. E.V.V. and C.Z. wrote the manuscript. All authors commented on the final draft of the manuscript and contributed to the analysis and interpretation of the data.

Corresponding authors

Correspondence to Bradley L. Pentelute or Stephen L. Buchwald.

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Competing interests

MIT has patents on the ligand used in this paper from which S.L.B. and former co-workers receive royalty payments.

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This file contains Supplementary Text and Data, Supplementary Tables 1-7, Supplementary Figures 1-16 and Supplementary References. (PDF 6807 kb)

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Vinogradova, E., Zhang, C., Spokoyny, A. et al. Organometallic palladium reagents for cysteine bioconjugation. Nature 526, 687–691 (2015). https://doi.org/10.1038/nature15739

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