2. Academic Validation
  3. (R)-PFI-2 is a potent and selective inhibitor of SETD7 methyltransferase activity in cells

(R)-PFI-2 is a potent and selective inhibitor of SETD7 methyltransferase activity in cells

  • Proc Natl Acad Sci U S A. 2014 Sep 2;111(35):12853-8. doi: 10.1073/pnas.1407358111.
Dalia Barsyte-Lovejoy 1,  Fengling Li 2,  Menno J Oudhoff 3,  John H Tatlock 4,  Aiping Dong 2,  Hong Zeng 2,  Hong Wu 2,  Spencer A Freeman 5,  Matthieu Schapira 6,  Guillermo A Senisterra 2,  Ekaterina Kuznetsova 2,  Richard Marcellus 7,  Abdellah Allali-Hassani 2,  Steven Kennedy 2,  Jean-Philippe Lambert 8,  Amber L Couzens 8,  Ahmed Aman 7,  Anne-Claude Gingras 9,  Rima Al-Awar 10,  Paul V Fish 11,  Brian S Gerstenberger 12,  Lee Roberts 13,  Caroline L Benn 14,  Rachel L Grimley 14,  Mitchell J S Braam 3,  Fabio M V Rossi 15,  Marius Sudol 16,  Peter J Brown 2,  Mark E Bunnage 13,  Dafydd R Owen 13,  Colby Zaph 17,  Masoud Vedadi 18,  Cheryl H Arrowsmith 19
Affiliations

Affiliations

  • 1 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; [email protected] [email protected] [email protected].
  • 2 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7;
  • 3 Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada V6T1Z3;
  • 4 Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, San Diego, CA 92121;
  • 5 Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC, Canada V6T1Z3;
  • 6 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada M5S 1A8;
  • 7 Drug Discovery Program, Ontario Institute for Cancer Research, Toronto, ON, Canada M5G 0A3;
  • 8 Centre for Systems Biology, Lunenfeld-Tanenbaum Research Institute, Toronto, ON, Canada M5G 1X5;
  • 9 Centre for Systems Biology, Lunenfeld-Tanenbaum Research Institute, Toronto, ON, Canada M5G 1X5; Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada M5S 1A8;
  • 10 Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada M5S 1A8; Drug Discovery Program, Ontario Institute for Cancer Research, Toronto, ON, Canada M5G 0A3;
  • 11 Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, Sandwich, Kent CT13 9NJ, United Kingdom;
  • 12 Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, Groton, CT 06340;
  • 13 Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, Cambridge, MA 02139;
  • 14 Neusentis Research Unit, Pfizer Worldwide Research and Development, Cambridge CB21 6GS, United Kingdom;
  • 15 Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada V6T1Z3; Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada V6T1Z3;
  • 16 Weis Center for Research, Geisinger Clinic, Danville, PA 17821;
  • 17 Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada V6T1Z3; Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada V6T1Z3; and.
  • 18 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada M5S 1A8; [email protected] [email protected] [email protected].
  • 19 Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; Princess Margaret Cancer Centre and Department of Medical Biophysics, University of Toronto, ON, Canada M5G 1L7 [email protected] [email protected] [email protected].
PMID: 25136132 DOI: 10.1073/pnas.1407358111
Abstract

SET domain containing (lysine methyltransferase) 7 (SETD7) is implicated in multiple signaling and disease related pathways with a broad diversity of reported substrates. Here, we report the discovery of (R)-PFI-2-a first-in-class, potent (Ki (app) = 0.33 nM), selective, and cell-active inhibitor of the methyltransferase activity of human SETD7-and its 500-fold less active enantiomer, (S)-PFI-2. (R)-PFI-2 exhibits an unusual cofactor-dependent and substrate-competitive inhibitory mechanism by occupying the substrate peptide binding groove of SETD7, including the catalytic lysine-binding channel, and by making direct contact with the donor methyl group of the cofactor, S-adenosylmethionine. Chemoproteomics experiments using a biotinylated derivative of (R)-PFI-2 demonstrated dose-dependent competition for binding to endogenous SETD7 in MCF7 cells pretreated with (R)-PFI-2. In murine embryonic fibroblasts, (R)-PFI-2 treatment phenocopied the effects of Setd7 deficiency on Hippo pathway signaling, via modulation of the transcriptional coactivator Yes-associated protein (YAP) and regulation of YAP target genes. In confluent MCF7 cells, (R)-PFI-2 rapidly altered YAP localization, suggesting continuous and dynamic regulation of YAP by the methyltransferase activity of SETD7. These data establish (R)-PFI-2 and related compounds as a valuable tool-kit for the study of the diverse roles of SETD7 in cells and further validate protein methyltransferases as a druggable target class.

Keywords

chemical biology; chemical probe; epigenetics.

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