Urea-Peptide Hybrids as VEGF-A165/NRP-1 Complex Inhibitors with Improved Receptor Affinity and Biological Properties.

Anna K Puszko,Karolina Pułka-Ziach,Olivier Hermine,Yves Lepelletier,Aleksandra Misicka,Gérard Hopfgartner,Rachel Rignault-Bricard,Piotr Sosnowski

doi:10.3390/ijms22010072

Abstract

Neuropilin-1 (NRP-1), the major co-receptor of vascular endothelial growth factor receptor-2 (VEGFR-2), may also independently act with VEGF-A165 to stimulate tumour growth and metastasis. Therefore, there is great interest in compounds that can block VEGF-A165/NRP-1 interaction. Peptidomimetic type inhibitors represent a promising strategy in the treatment of NRP-1-related disorders. Here, we present the synthesis, affinity, enzymatic stability, molecular modeling and in vitro binding evaluation of the branched urea–peptide hybrids, based on our previously reported Lys(hArg)-Dab-Oic-Arg active sequence, where the Lys(hArg) branching has been modified by introducing urea units to replace the peptide bond at various positions. One of the resulting hybrids increased the affinity of the compound for NRP-1 more than 10-fold, while simultaneously improving resistance for proteolytic stability in serum. In addition, ligand binding to NRP-1 induced rapid protein stock exocytotic trafficking to the plasma membrane in breast cancer cells. Examined properties characterize this compound as a good candidate for further development of VEGF165/NRP-1 inhibitors.

Highlights

The most important member from the vascular endothelial growth factors (VEGFs) family is the VEGF-A165 isoform, which has been shown to play major roles in physiological and pathological angiogenesis [1,2]
It affects vascular permeability through binding to type III tyrosine kinase receptors from the vascular endothelial growth factor receptors (VEGF-R) family: VEGF-R1 and VEGF-R2 [1,2,3,4,5]. Another significant receptor for VEGF-A165 is neuropilin-1 (NRP-1), expressed on endothelial cells and responsible for enhancing the VEGF-A165 /VEGF-R2 signaling as a co-receptor, thereby increasing endothelial cell proliferation and migration and promoting angiogenesis [6,7,8,9]
Our previous studies showed that the substitution of Lys-ε-hArg amide bond by urea group and replacement of hArg with shorter Arg side chain made peptidomimetics more resistant to enzymatic cleavage, but it affected the affinity for NRP-1

Summary

Introduction

The most important member from the vascular endothelial growth factors (VEGFs) family is the VEGF-A165 isoform, which has been shown to play major roles in physiological and pathological angiogenesis [1,2] It affects vascular permeability through binding to type III tyrosine kinase receptors from the vascular endothelial growth factor receptors (VEGF-R) family: VEGF-R1 and VEGF-R2 [1,2,3,4,5]. Another significant receptor for VEGF-A165 is neuropilin-1 (NRP-1), expressed on endothelial cells and responsible for enhancing the VEGF-A165 /VEGF-R2 signaling as a co-receptor, thereby increasing endothelial cell proliferation and migration and promoting angiogenesis [6,7,8,9]. Designed inhibitors include small molecules [22,23,24,25,26,27,28,29] and linear [30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46]

Objectives

Methods

Results

Conclusion