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Synthesis, Characterization, and Biological Evaluation of a Dual-Action Ligand Targeting αvβ3 Integrin and VEGF Receptors.

Zanella S, Mingozzi M, Dal Corso A, Fanelli R, Arosio D, Cosentino M, Schembri L, Marino F, De Zotti M, Formaggio F, Pignataro L, Belvisi L, Piarulli U, Gennari C - ChemistryOpen (2015)

Bottom Line: In vitro binding assays on isolated integrin αVβ3 and VEGFR-1 showed that the dual-action conjugate retains a good level of affinity for both its target receptors, although with one order of magnitude (10/20 times) decrease in potency.The dual-action ligand strongly inhibited the VEGF-induced morphogenesis in Human Umbilical Vein Endothelial Cells (HUVECs).Remarkably, its efficiency in preventing the formation of new blood vessels was similar to that of the original individual ligands, despite the worse affinity towards integrin αVβ3 and VEGFR-1.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Chimica, Università degli Studi di Milano Via C. Golgi 19, 20133, Milan, Italy.

ABSTRACT
A dual-action ligand targeting both integrin αVβ3 and vascular endothelial growth factor receptors (VEGFRs), was synthesized via conjugation of a cyclic peptidomimetic αVβ3 Arg-Gly-Asp (RGD) ligand with a decapentapeptide. The latter was obtained from a known VEGFR antagonist by acetylation at the Lys13 side chain. Functionalization of the precursor ligands was carried out in solution and in the solid phase, affording two fragments: an alkyne VEGFR ligand and the azide integrin αVβ3 ligand, which were conjugated by click chemistry. Circular dichroism studies confirmed that both the RGD and VEGFR ligand portions of the dual-action compound substantially adopt the biologically active conformation. In vitro binding assays on isolated integrin αVβ3 and VEGFR-1 showed that the dual-action conjugate retains a good level of affinity for both its target receptors, although with one order of magnitude (10/20 times) decrease in potency. The dual-action ligand strongly inhibited the VEGF-induced morphogenesis in Human Umbilical Vein Endothelial Cells (HUVECs). Remarkably, its efficiency in preventing the formation of new blood vessels was similar to that of the original individual ligands, despite the worse affinity towards integrin αVβ3 and VEGFR-1.

No MeSH data available.


Related in: MedlinePlus

CD spectra of peptide 4 in water, 2,2,2-trifluoroethanol (TFE) and methanol (MeOH) (0.1 mm).
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fig04: CD spectra of peptide 4 in water, 2,2,2-trifluoroethanol (TFE) and methanol (MeOH) (0.1 mm).

Mentions: The circular dichroism (CD) spectrum of peptide 4 measured in water (Figure 4) is consistent with that published by D′Andrea and co-workers for the parent peptide 3.14 The position and the intensities of the two negative maxima indicate the presence of a relevant population of α-helical structures. However, the negative maximum of the amide π→π* electronic transition, located at 203 nm, is significantly blue-shifted with respect to the canonical position (208 nm) of an α-helical conformation. This finding, together with the intensity decrease of the band at about 222 nm, can be safely assigned to a non-negligible participation of unordered conformations, also called “polyproline type-II”. We extended our analysis to two additional solvents, namely trifluoroethanol (TFE) and methanol, roughly mimicking the hydrophobic biological environment in which the peptide would probably display its action.22 These CD spectra (Figure 4) are characterized by two negative maxima typical of an α-helical conformation. This increase in the helical content, moving from water to the alcoholic solvents, clearly indicates that peptide 4 is not very rigid, as its 3 D-structure is highly dependent on the environment. The same is true for the parent peptide 3, whose CD spectra undergo a parallel modification in the cited solvents (see the Supporting Information).


Synthesis, Characterization, and Biological Evaluation of a Dual-Action Ligand Targeting αvβ3 Integrin and VEGF Receptors.

Zanella S, Mingozzi M, Dal Corso A, Fanelli R, Arosio D, Cosentino M, Schembri L, Marino F, De Zotti M, Formaggio F, Pignataro L, Belvisi L, Piarulli U, Gennari C - ChemistryOpen (2015)

CD spectra of peptide 4 in water, 2,2,2-trifluoroethanol (TFE) and methanol (MeOH) (0.1 mm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608532&req=5

fig04: CD spectra of peptide 4 in water, 2,2,2-trifluoroethanol (TFE) and methanol (MeOH) (0.1 mm).
Mentions: The circular dichroism (CD) spectrum of peptide 4 measured in water (Figure 4) is consistent with that published by D′Andrea and co-workers for the parent peptide 3.14 The position and the intensities of the two negative maxima indicate the presence of a relevant population of α-helical structures. However, the negative maximum of the amide π→π* electronic transition, located at 203 nm, is significantly blue-shifted with respect to the canonical position (208 nm) of an α-helical conformation. This finding, together with the intensity decrease of the band at about 222 nm, can be safely assigned to a non-negligible participation of unordered conformations, also called “polyproline type-II”. We extended our analysis to two additional solvents, namely trifluoroethanol (TFE) and methanol, roughly mimicking the hydrophobic biological environment in which the peptide would probably display its action.22 These CD spectra (Figure 4) are characterized by two negative maxima typical of an α-helical conformation. This increase in the helical content, moving from water to the alcoholic solvents, clearly indicates that peptide 4 is not very rigid, as its 3 D-structure is highly dependent on the environment. The same is true for the parent peptide 3, whose CD spectra undergo a parallel modification in the cited solvents (see the Supporting Information).

Bottom Line: In vitro binding assays on isolated integrin αVβ3 and VEGFR-1 showed that the dual-action conjugate retains a good level of affinity for both its target receptors, although with one order of magnitude (10/20 times) decrease in potency.The dual-action ligand strongly inhibited the VEGF-induced morphogenesis in Human Umbilical Vein Endothelial Cells (HUVECs).Remarkably, its efficiency in preventing the formation of new blood vessels was similar to that of the original individual ligands, despite the worse affinity towards integrin αVβ3 and VEGFR-1.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Chimica, Università degli Studi di Milano Via C. Golgi 19, 20133, Milan, Italy.

ABSTRACT
A dual-action ligand targeting both integrin αVβ3 and vascular endothelial growth factor receptors (VEGFRs), was synthesized via conjugation of a cyclic peptidomimetic αVβ3 Arg-Gly-Asp (RGD) ligand with a decapentapeptide. The latter was obtained from a known VEGFR antagonist by acetylation at the Lys13 side chain. Functionalization of the precursor ligands was carried out in solution and in the solid phase, affording two fragments: an alkyne VEGFR ligand and the azide integrin αVβ3 ligand, which were conjugated by click chemistry. Circular dichroism studies confirmed that both the RGD and VEGFR ligand portions of the dual-action compound substantially adopt the biologically active conformation. In vitro binding assays on isolated integrin αVβ3 and VEGFR-1 showed that the dual-action conjugate retains a good level of affinity for both its target receptors, although with one order of magnitude (10/20 times) decrease in potency. The dual-action ligand strongly inhibited the VEGF-induced morphogenesis in Human Umbilical Vein Endothelial Cells (HUVECs). Remarkably, its efficiency in preventing the formation of new blood vessels was similar to that of the original individual ligands, despite the worse affinity towards integrin αVβ3 and VEGFR-1.

No MeSH data available.


Related in: MedlinePlus