Branched pentapeptides as potent inhibitors of the vascular endothelial growth factor 165 binding to Neuropilin-1: Design, synthesis and biological activity

Tymecka, Dagmara; Puszko, Anna K.; Lipiński, Piotr F. J.; Fedorczyk, Bartlomiej; Wileńska, Beata; Sura, Karolina; Perret, G; Misicka, Aleksandra

doi:10.1016/j.ejmech.2018.08.083

Cited by 23 publications

(39 citation statements)

References 49 publications

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“…[ 108 ] Based on these findings, the group of Misicka has developed branched pentapeptides of the type Lys(Har)-Xaa-Xaa-Arg (Har = homoarginine) with submicromolar IC 50 values, for example 0.3 μM for Xaa-Xaa= Pro-Ala ( NV1 ). However, they observed enzymatic cleavage of the peptides, which led to a low blood plasma stability in vitro [ 109 ]. To overcome the peptide’s instability, the authors turned their attention to 1,4-disubstituted 1,2,3-triazoles as a stable amide bond surrogate.…”

Section: Applicationsmentioning

confidence: 99%

“…However, the in vitro plasma stability results revealed that these two analogues are significantly more stable. After 48 h, 70% of compound NV2 and >90% of compound NV3 were still intact (half-lives not determined), whereas the previously reported compound NV1 possess a half-life of only 39 min [ 109 ]. Further modification of NV2 at the N-terminus was attempted to optimise the inhibition of NRP-1/VEGF 165 interaction but efforts remained unsuccessful.…”

Section: Applicationsmentioning

confidence: 99%

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1,4-Disubstituted 1,2,3-Triazoles as Amide Bond Surrogates for the Stabilisation of Linear Peptides with Biological Activity

2020

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Peptides represent an important class of biologically active molecules with high potential for the development of diagnostic and therapeutic agents due to their structural diversity, favourable pharmacokinetic properties, and synthetic availability. However, the widespread use of peptides and conjugates thereof in clinical applications can be hampered by their low stability in vivo due to rapid degradation by endogenous proteases. A promising approach to circumvent this potential limitation includes the substitution of metabolically labile amide bonds in the peptide backbone by stable isosteric amide bond mimetics. In this review, we focus on the incorporation of 1,4-disubstituted 1,2,3-triazoles as amide bond surrogates in linear peptides with the aim to increase their stability without impacting their biological function(s). We highlight the properties of this heterocycle as a trans-amide bond surrogate and summarise approaches for the synthesis of triazole-containing peptidomimetics via the Cu(I)-catalysed azide-alkyne cycloaddition (CuAAC). The impacts of the incorporation of triazoles in the backbone of diverse peptides on their biological properties such as, e.g., blood serum stability and affinity as well as selectivity towards their respective molecular target(s) are discussed.

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Section: Applicationsmentioning

confidence: 99%

Section: Applicationsmentioning

confidence: 99%

1,4-Disubstituted 1,2,3-Triazoles as Amide Bond Surrogates for the Stabilisation of Linear Peptides with Biological Activity

2020

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“…Figure 4E). Thus, all of compounds are able to partially mimic the terminal carboxyl contacts that are considered to be critical for anchoring the terminal Arg of CendR peptides 12,31,[38][39][40][41][42][44][45][46][47][48][49][50][51][52][53] (Figure 1C) or known small molecule mimetics and inhibitors that contain a terminal guanidyl from Arg moiety and carboxylic group 54,61,62 . Figure 4F).…”

Section: Series Analysis and Docked Binding Modesmentioning

confidence: 99%

“…Due to its role in cancer, NRP-1 has been a target for drug design for over 20 years. During this time, discovery efforts have focused on development of NRP-1 antibody therapies 9,[28][29][30][31][32][33] , including a recent dual-specificity antibody to VEGFA and NRP-1 32,33 , peptides that target transmembrane domain interactions 11,[34][35][36][37] or the CendR interaction site 12,31,[38][39][40][41][42][43][44][45][46][47][48][49][50][51][52][53] , as well as small-molecules that target the CendR site [54][55][56][57][58][59][60][61] . With one exception 43 , the CendR site targeting peptides contain a CendR motif, even those which are cyclical 39,45,46,49 or branched 51,53 .…”

Section: Introductionmentioning

confidence: 99%

In silico identification and validation of inhibitors of the interaction between neuropilin receptor 1 and SARS-CoV-2 Spike protein

Perez‐Miller

Pátek²,

Moutal

et al. 2020

Preprint

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Neuropilin-1 (NRP-1) is a multifunctional transmembrane receptor for ligands that affect developmental axonal growth and angiogenesis. In addition to a role in cancer, NRP-1 is a reported entry point for several viruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causal agent of coronavirus disease 2019 (COVID-19). The furin cleavage product of SARS-CoV-2 Spike protein takes advantage of the vascular endothelial growth factor A (VEGF-A) binding site on NRP-1 which accommodates a polybasic stretch ending in a C-terminal arginine. This site has long been a focus of drug discovery efforts for cancer therapeutics. We recently showed that interruption of the VEGF-A/NRP-1 signaling pathway ameliorates neuropathic pain and hypothesize that interference of this pathway by SARS-CoV-2 spike protein interferes with pain signaling. Here, we report hits from a small molecule and natural product screen of nearly 0.5 million compounds targeting the VEGF-A binding site on NRP-1. We identified nine chemical series with lead- or drug-like physico-chemical properties. Using an ELISA, we demonstrate that six compounds disrupt VEGF-A-NRP-1 binding more effectively than EG00229, a known NRP-1 inhibitor. Secondary validation in cells revealed that almost all tested compounds inhibited VEGF-A triggered VEGFR2 phosphorylation. Two compounds displayed robust inhibition of a recombinant vesicular stomatitis virus protein that utilizes the SARS-CoV-2 Spike for entry and fusion. These compounds represent a first step in a renewed effort to develop small molecule inhibitors of the VEGF-A/NRP-1 signaling for the treatment of neuropathic pain and cancer with the added potential of inhibiting SARS-CoV-2 virus entry.

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“…All of these interactions may play a crucial role in the formation of the ligand-receptor complex.Among the antiangiogenic molecules, a groundbreaking discovery was the A7R heptapeptide, isolated using phage display library screening, which in vivo decreases breast cancer angiogenesis and growth [19][20][21]. Based on the A7R peptide, which showed the ability to inhibit VEGF-A 165 /NRP-1 complex formation with a high IC 50 value (IC 50 = 80 µM) [21], and respecting CendR rules, we previously obtained short-branched pentapeptides, with a general sequence Lys(hArg)-AA 2 -AA 3 -Arg, that have reduced IC 50 values (IC 50 = 2-20 µM) [37,38]. In the present study, we decided to examine how an additional Cys-Asp (CD) fragment (present at the N-terminus of the C-terminal hexapeptide of VEGF-A 165 -CDKPRR) or Cys (C) residue will influence affinity for NRP-1 of the best analogue of our branched peptides, namely Lys(hArg)-Dap-Dhp-Arg.…”

mentioning

confidence: 99%

Does Cysteine Rule (CysR) Complete the CendR Principle? Increase in Affinity of Peptide Ligands for NRP-1 Through the Presence of N-Terminal Cysteine

et al. 2020

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The structure-activity relationship of branched H-Lys(hArg)-Dab-Dhp-Arg-OH sequence analogues, modified with Cys-Asp or Cys at N-terminal amino acids (Lys, hArg), in VEGF-A 165 /Neuropilin-1 complex inhibition is presented. The addition of Cys residue led to a 100-fold decrease in the IC 50 value, compared to the parent peptide. The change occurred regardless of coupling Cys to the free N-terminal amino group present in the main or the side chain. A few analogues extended by the attachment of Cys at the N-terminus of several potent NRP-1 peptide ligands documented in the literature are also presented. In all studied cases, the enhancement of inhibitory properties after the addition of Cys at the N-terminus is observed. It is particularly evident for the tetrapeptide derived from the C-terminus of VEGF-A 165 (KPRR), suggesting that extending the K/RXXK/R motif (CendR) with the Cys moiety can significantly improve affinity to NRP-1 of CendR peptides.Biomolecules 2020, 10, 448 2 of 11 development in cancerous tumors and that its expression on the surface of cancerous cells is increased and often associated with poor prognosis [10][11][12][13]. These results indicate that searching for new NRP-1 inhibitors of pathological angiogenesis, including functional peptides, could play a crucial role in drug development (e.g., in oncology).The VEGF-A 165 fragment encoded by exons 7 and 8 is mainly responsible for protein interaction with NRP-1 [14-17]. The binding pocket of the NRP-1 b1 domain is shallow, and only the C-terminal Arg of the ligand is able to directly enter into it [6,18]. Screening of phage libraries led to the identification of peptides possessing a R/KXXR/K consensus motif embedded in their terminal sequence, which showed a high binding affinity to NRP-1 [18]. As with VEGF-A 165 , the C-terminal Arg (or rarely, Lys) was important for NRP-1-binding. Blocking the free carboxyl group of this Arg residue by amidation or adding another amino acid eliminated ligand binding to the receptor. The strict requirements of the R/KXXR/K C-terminal structural motif in NRP-1 peptidic ligands is called the C-end rule (CendR) [19]. The CendR fragment is present in the structure of active peptidic NRP-1 ligands described in the literature, e.g., heptapeptide A7R (ATWLPPR) [19][20][21], shorter analogues of A7R [22], tuftsin [23], and penetrating iRGD peptide [24] as well as the C-terminus of VEGF-A 165 (CDKPRR) encoded by the exon 8 and multiple viruses' capsid proteins, e.g., human T-cell lymphotropic virus type 1 (HTLV-1) [25,26] and Epstein-Barr virus (EBV) [27].Another characteristic feature of the VEGF-A 165 structure is the presence of eight invariant cysteine (Cys) residues associated with intermolecular and intramolecular disulfide bonds [28]. The N-terminal fragment of the VEGF-A 165 domain ERTCDKPRR encoded by exons 7 and 8, as well as its shorter analogue CDKPRR, showed nearly 100% inhibition of the formation of the VEGF-A 165 /NRP-1 complex at a concentration of 100 µM [29]. The replacement of Cys with Ser at posit...

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Branched pentapeptides as potent inhibitors of the vascular endothelial growth factor 165 binding to Neuropilin-1: Design, synthesis and biological activity

Cited by 23 publications

References 49 publications

1,4-Disubstituted 1,2,3-Triazoles as Amide Bond Surrogates for the Stabilisation of Linear Peptides with Biological Activity

1,4-Disubstituted 1,2,3-Triazoles as Amide Bond Surrogates for the Stabilisation of Linear Peptides with Biological Activity

In silico identification and validation of inhibitors of the interaction between neuropilin receptor 1 and SARS-CoV-2 Spike protein

Does Cysteine Rule (CysR) Complete the CendR Principle? Increase in Affinity of Peptide Ligands for NRP-1 Through the Presence of N-Terminal Cysteine

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