Macrocyclized Extended Peptides: Inhibiting the Substrate-Recognition Domain of Tankyrase
Xu, W Lau, Y.H Fischer, G Tan, Y.S Chattopadhyay, A De La Roche, M Hyvönen, M Verma, C Spring, D.R Itzhaki, L.S
Xu, W
Lau, Y.H
Fischer, G
Tan, Y.S
Chattopadhyay, A
De La Roche, M
Hyvönen, M
Spring, D.R
Itzhaki, L.S
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Alternative Title
Abstract
We report a double-click macrocyclization approach for the design of constrained peptide inhibitors having non-helical or extended conformations. Our targets are the tankyrase proteins (TNKS), poly(ADP-ribose) polymerases (PARP) that regulate Wnt signaling by targeting Axin for degradation. TNKS are deregulated in many different cancer types, and inhibition of TNKS therefore represents an attractive therapeutic strategy. However, clinical development of TNKS-specific PARP catalytic inhibitors is challenging due to off-target effects and cellular toxicity. We instead targeted the substrate-recognition domain of TNKS, as it is unique among PARP family members. We employed a two-component strategy, allowing peptide and linker to be separately engineered and then assembled in a combinatorial fashion via click chemistry. Using the consensus substrate-peptide sequence as a starting point, we optimized the length and rigidity of the linker and its position along the peptide. Optimization was further guided by high-resolution crystal structures of two of the macrocyclized peptides in complex with TNKS. This approach led to macrocyclized peptides with submicromolar affinities for TNKS and high proteolytic stability that are able to disrupt the interaction between TNKS and Axin substrate and to inhibit Wnt signaling in a dose-dependent manner. The peptides therefore represent a promising starting point for a new class of substrate-competitive inhibitors of TNKS with potential for suppressing Wnt signaling in cancer. Moreover, by demonstrating the application of the double-click macrocyclization approach to non-helical, extended, or irregularly structured peptides, we greatly extend its potential and scope, especially given the frequency with which such motifs mediate protein-protein interactions. © 2017 American Chemical Society.
Keywords
Crystal structure, Diseases, Clinical development, Dose-dependent manner, High resolution crystal structure, Poly(ADP-ribose) polymerase, Protein-protein interactions, Proteolytic stability, Sub-micromolar affinity, Substrate recognition, Peptides, alkynyl group, ankyrin, axin, macrocyclic compound, nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase, peptide, protein MDM2, protein p53, tankyrase, telomeric repeat binding factor 1, urea derivative, enzyme inhibitor, macrocyclic compound, peptide, tankyrase, amino acid sequence, amino terminal sequence, Article, circular dichroism, click chemistry, competitive inhibition, consensus sequence, controlled study, cross linking, crystal structure, cyclization, cytotoxicity, dimerization, dissociation constant, double click chemistry, fluorescence polarization, human, human cell, hydrogen bond, infrared spectrometry, liquid chromatography-mass spectrometry, M phase cell cycle checkpoint, macrocyclization, malignant neoplasm, molecular dynamics, molecular recognition, protein conformation, protein degradation, protein interaction, stoichiometry, telomere length, thermodynamics, wild type, Wnt signaling pathway, antagonists and inhibitors, chemical structure, chemistry, isolation and purification, metabolism, synthesis, X ray crystallography, Click Chemistry, Crystallography, X-Ray, Enzyme Inhibitors, Humans, Macrocyclic Compounds, Molecular Dynamics Simulation, Molecular Structure, Peptides, Tankyrases, Thermodynamics
Source Title
Journal of the American Chemical Society
Publisher
American Chemical Society
Series/Report No.
Collections
Rights
Attribution 4.0 International
Date
2017
DOI
10.1021/jacs.6b10234
Type
Article