Rational design of dual-domain binding inhibitors for N-acetylgalactosamine transferase 2 with improved selectivity over the T1 and T3 isoforms

Ismael Compañón, Collin J. Ballard, Erandi Lira-Navarrete, Tanausú Santos, Serena Monaco, Juan C. Muñoz-García, Ignacio Delso, Jesus Angulo, Thomas A. Gerken, Katrine T. Schjoldager, Henrik Clausen, Tomás Tejero, Pedro Merino, Francisco Corzana, Ramon Hurtado-Guerrero, Mattia Ghirardello (Lead Author)

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Abstract

The GalNAc-transferase (GalNAc-T) family, consisting of 20 isoenzymes, regulates the O-glycosylation process of mucin glycopeptides by transferring GalNAc units to serine/threonine residues. Dysregulation of specific GalNAc-Ts is associated with various diseases, making these enzymes attractive targets for drug development. The development of inhibitors is key to understanding the implications of GalNAc-Ts in human diseases. However, developing selective inhibitors for individual GalNAc-Ts represents a major challenge due to shared structural similarities among the isoenzymes and some degree of redundancy among the natural substrates. Herein, we report the development of a GalNAc-T2 inhibitor with higher potency compared to those of the T1 and T3 isoforms. The most promising candidate features bivalent GalNAc and thiophene moieties on a peptide chain, enabling binding to both the lectin and catalytic domains of the enzyme. The binding mode was confirmed by competitive saturation transfer difference NMR experiments and validated through molecular dynamics simulations. The inhibitor demonstrated an IC50 of 21.4 μM for GalNAc-T2, with 8- and 32-fold higher selectivity over the T3 and T1 isoforms, respectively, representing a significant step forward in the synthesis of specific GalNAc-T inhibitors tailored to the unique structural features of the targeted isoform.

Original languageEnglish
Pages (from-to)3649-3656
Number of pages8
JournalJACS Au
Volume4
Issue number9
Early online date11 Sep 2024
DOIs
Publication statusPublished - 23 Sep 2024

Keywords

  • GalNAc
  • glycopeptide
  • glycosyltransferase
  • inhibitor
  • molecular dynamics
  • N-acetylgalactosamine transferase
  • STD NMR

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