Potent and selective SETDB1 covalent negative allosteric modulator reduces methyltransferase activity in cells

• DOI: 10.1038/s41467-025-57005-3
• Publication Date: 2025-02-24T06:07:57Z
• Authors: Mélanie Uguen; Devan J. Shell; Madhushika Silva; Yu Deng; Fengling Li; Magdalena M. Szew...; Ka Yang; Yani Zhao; Michael A. Stashko; Jacqueline L. Nor...; Jarod M. Waybright; Serap Beldar; Justin M. Rectenwald; Angie L. Mordant; Thomas S. Webb; Laura E. Herring; Cheryl H. Arrowsmith; Suzanne Ackloo; Steven P. Gygi; Robert K. McGinty; Dalia Barsyte-Lov...; Pengda Liu; Levon Halabelian; Lindsey I. James; Kenneth H. Pearce; Stephen V. Frye
• Keywords: Allosteric modulator

A promising drug target, SETDB1, is a dual methyl-lysine (Kme) reader and methyltransferase implicated in cancer neurodegenerative disease progression. To help understand the role of triple Tudor domain (3TD) its Kme reader, we first identify low micromolar potency small molecule ligand, UNC6535, which occupies simultaneously both TD2 TD3 binding sites. Further optimization leads to discovery UNC10013, covalent 3TD ligand targeting Cys385 SETDB1. UNC10013 potent with kinact/KI 1.0 × 106 M−1s−1 demonstrates proteome-wide selectivity. In cells, negative allosteric modulation SETDB1-mediated Akt methylation occurs after treatment UNC10013. Therefore, potent, selective, cell-active for demonstrating modulator properties making it tool study biological SETDB1 Design cysteine-targeting analogs reversible led high demonstrated inhibition approach therapeutics.