TY - JOUR
T1 - Rational design of dual-domain binding inhibitors for N-acetylgalactosamine transferase 2 with improved selectivity over the T1 and T3 isoforms
AU - Compañón, Ismael
AU - Ballard, Collin J.
AU - Lira-Navarrete, Erandi
AU - Santos, Tanausú
AU - Monaco, Serena
AU - Muñoz-García, Juan C.
AU - Delso, Ignacio
AU - Angulo, Jesus
AU - Gerken, Thomas A.
AU - Schjoldager, Katrine T.
AU - Clausen, Henrik
AU - Tejero, Tomás
AU - Merino, Pedro
AU - Corzana, Francisco
AU - Hurtado-Guerrero, Ramon
AU - Ghirardello, Mattia
N1 - Funding Information: Agencia Estatal de Investigación (AEI, PID2021-127622OB-I00 and PDC2022-133725-C21 to F.C., PID2022-136362NB-I00 to R.H.-G., PID2022137973NB-I00 to P.M.), Universidad de La Rioja (REGI22/16 Project), and Gobierno de Aragón (E34_R17 and LMP58_18) with FEDER (2014–2020) funds for ‘Building Europe from Aragón’ for financial support (to R.H.-G.). The support of the National Institutes of Health grant R01-GM113535 (to T.A.G.) is also acknowledged. M.G. and I.D. acknowledge the Marie Skłodowska-Curie fellowship program (grant agreement nos. 101034288 (M.G.) and 890779 (I.D.)) for financial support. J.A. and S.M. acknowledge support of BBSRC, grant BB/P010660/1. J.A. and J.C.M.-G. also acknowledge funding from the grant AEI/10.13039/501100011033/PID2022-142879NB-I00, cofunded by the European Regional Development Fund (ERDF), “A way of making Europe”, and T.S. thanks MICIU/AEI /10.13039/501100011033 and the European Union NextGenerationEU/PRTR for his Juan de la Cierva contract, JDC2022-048607-I.
PY - 2024/9/23
Y1 - 2024/9/23
N2 - 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.
AB - 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.
KW - GalNAc
KW - glycopeptide
KW - glycosyltransferase
KW - inhibitor
KW - molecular dynamics
KW - N-acetylgalactosamine transferase
KW - STD NMR
UR - http://www.scopus.com/inward/record.url?scp=85203833745&partnerID=8YFLogxK
U2 - 10.1021/jacsau.4c00633
DO - 10.1021/jacsau.4c00633
M3 - Article
AN - SCOPUS:85203833745
VL - 4
SP - 3649
EP - 3656
JO - JACS Au
JF - JACS Au
SN - 2691-3704
IS - 9
ER -