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Protocols for Synthesis of SNIPERs and the Methods to Evaluate the Anticancer Effects

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Targeted Protein Degradation

Abstract

Inducing degradation of undruggable target proteins by the use of chimeric small molecules, represented by proteolysis-targeting chimeras, is a promising strategy for drug development. We developed a series of chimeric molecules, termed “specific and nongenetic inhibitor of apoptosis protein (IAP)-dependent protein erasers” (SNIPERs) that recruit IAP ubiquitin ligases to induce degradation of target proteins. SNIPERs also induce degradation of some IAPs, including cIAP1 and XIAP, which are antiapoptotic proteins that are overexpressed in many cancers. Such protein degraders have unique properties that could be especially useful in cancer therapy. This chapter describes (1) the design and synthesis of SNIPER compounds, (2) the methods used for the detection of target protein degradation and ubiquitylation, and (3) the protocol to evaluate the antitumor activity of SNIPER.

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Acknowledgments

This study was supported by MEXT/JSPS KAKENHI (grants JP19K07724 and JP16K18444 to Y.T., JP19K16333 to G.T., JP18K07311 to N.S., JP18K06567 to N.O., JP17K08385 to Y.D., JP16H05090, and JP18H05502 to M.N.) and by AMED (grants JP17cm0106522j0002 to N.S., JP19cm0106136, JP19ak0101073 to N.O., JP19ak0101073j0603, and JP19im0210616j0002 to M.N.). We are grateful to Journal of Biological Chemistry and the American Society for Biochemistry and Molecular Biology (ASBMB) for allowing the reproduction of Figs. 5, 6, and 7, which were originally published in the Journal of Biological Chemistry [8] (Ohoka N, Okuhira K, Ito M et al. In vivo knockdown of pathogenic proteins via specific and nongenetic inhibitor of apoptosis protein (IAP)-dependent protein erasers (SNIPERs). J Biol Chem. 2017; 11:4556–4570. © the American Society for Biochemistry and Molecular Biology.) We also thank Marla Brunker, from Edanz Group (www.edanzediting.com/ac), for editing a draft of this manuscript.

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Correspondence to Mikihiko Naito .

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Tsukumo, Y. et al. (2021). Protocols for Synthesis of SNIPERs and the Methods to Evaluate the Anticancer Effects. In: Cacace, A.M., Hickey, C.M., Békés, M. (eds) Targeted Protein Degradation. Methods in Molecular Biology, vol 2365. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1665-9_18

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  • DOI: https://doi.org/10.1007/978-1-0716-1665-9_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1664-2

  • Online ISBN: 978-1-0716-1665-9

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