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N-tert-butyloxycarbonyl-Phe-Leu-Phe-Leu-Phe (BOC2) inhibits the angiogenic activity of heparin-binding growth factors

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Abstract

The peptides N-tert-butyloxycarbonyl-Phe-Leu-Phe-Leu-Phe (BOC2) and BOC-Met-Leu-Phe (BOC1) are widely used antagonists of formyl peptide receptors (FPRs), BOC2 acting as an FPR1/FPR2 antagonist whereas BOC1 inhibits FPR1 only. Extensive investigations have been performed by using these FPR antagonists as a tool to assess the role of FPRs in physiological and pathological conditions. Based on previous observations from our laboratory, we assessed the possibility that BOC2 may exert also a direct inhibitory effect on the angiogenic activity of vascular endothelial growth factor-A (VEGF-A). Our data demonstrate that BOC2, but not BOC1, inhibits the angiogenic activity of heparin-binding VEGF-A165 with no effect on the activity of the non-heparin-binding VEGF-A121 isoform. Endothelial cell-based bioassays, surface plasmon resonance analysis, and computer modeling indicate that BOC2 may interact with the heparin-binding domain of VEGF-A165, thus competing for heparin interaction and preventing the binding of VEGF-A165 to tyrosine kinase receptor VEGFR2, its phosphorylation and downstream signaling. In addition, BOC2 inhibits the interaction of a variety of heparin-binding angiogenic growth factors with heparin, including fibroblast growth factor 2 (FGF2) whose angiogenic activity is blocked by the compound. Accordingly, BOC2 suppresses the angiogenic potential of human tumor cell lines that co-express VEGF-A and FGF2. Thus, BOC2 appears to act as a novel multi-heparin-binding growth factor antagonist. These findings caution about the interpretation of FPR-focusing experimental data obtained with this compound and set the basis for the design of novel BOC2-derived, FPR independent multi-target angiogenesis inhibitors.

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Acknowledgements

The authors wish to thank S. Matarazzo for technical support. This work was supported in part by Associazione Italiana per la Ricerca sul Cancro (IG AIRC Grant No. 18493) to M. P.; S. R. was supported by an AIRC fellowship.

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Authors and Affiliations

  1. Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy

    Imtiaz M. Nawaz, Paola Chiodelli, Sara Rezzola, Giuseppe Paganini, Michela Corsini & Marco Presta

  2. Department of Food and Drug, University of Parma, Parma, Italy

    Alessio Lodola, Alessio Di Ianni & Marco Mor

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  1. Imtiaz M. Nawaz
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Correspondence to Marco Presta.

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Nawaz, I.M., Chiodelli, P., Rezzola, S. et al. N-tert-butyloxycarbonyl-Phe-Leu-Phe-Leu-Phe (BOC2) inhibits the angiogenic activity of heparin-binding growth factors. Angiogenesis 21, 47–59 (2018). https://doi.org/10.1007/s10456-017-9581-6

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