Synthesis of Modified RGD‐Based Peptides and Their in vitro Activity

Arg‐Gly‐Asp (RGD) peptides represent the most outstanding recognition motif involved in cell adhesion that binds to the αvβ3 integrin, which has been targeted for cancer therapy. Various RGD‐containing peptides and peptidomimetics have been designed and synthesized to selectively inhibit this integrin. In this study, the synthesis of RGD‐based peptides through the incorporation of the short bioactive peptide Phe‐Ala‐Lys‐Leu‐Phe (FAKLF) at the C and N termini of RGD has been achieved by using a solid‐phase peptide synthesis approach. The peptides were purified by means of preparative RP‐HPLC and their structures were confirmed through HRMS (ESI). The MTT assay revealed that the RGD and FAKLF peptides inhibited the proliferation of human umbilical vein endothelial cells (HUVECs) in a dose‐dependent manner, with IC50 values of 3000 and 500 ng mL−1, respectively. Interestingly, a drastic improvement was observed in the antiproliferative activity of the combined structures of the FAKLFRGD and RGDFAKLF peptides, leading to IC50 values of 200 and 136.7 ng mL−1, respectively. Meanwhile, based on apoptosis results, the potential of peptides to induce apoptosis, in accordance with their antiproliferative activity, indicated that the RGD and FAKLF peptides, and the peptides synthesized based on their combinations induced cell apoptosis in a dose‐dependent manner followed by inhibition of the proliferation of endothelial cells. Moreover, the incorporation of d‐leucine increased the induction of apoptosis by these peptides. Programmed to die: The synthesis of peptides containing the RGD and FAKLF motifs are described. An in vitro study shows that peptides containing a combination of these fragments, FAKLFRGD and RGDFAKLF, have greater suppressing activity toward endothelial cells. Moreover, the incorporation of d‐leucine increases the induction of apoptosis by these peptides.