Fabrication of self-assembled peptide nanoparticles for in vitro assessment of cell apoptosis pathway and in vivo therapeutic efficacy

Near-infrared fluorescent (NIRF) dye-coupled self-assembled RGD-linked proapoptotic peptide nanoparticles have been synthesized with spherical shape and size ~ 30–40 nm diameters. The peptide sequence was coupled with cyanine 5.5 probe as NIRF-dye to introduce optical imaging properties and pH-dependent method was used to design Cy5.5 coupled self-assembled peptide nanoparticles ( f -SAPNs). This nanoprobe has the ability to target α v β 3 -integrin receptor overexpressed on cancer cell’s surface with improved internalization capabilities into the mitochondria. The in situ study showed that this peptide sequence has potential to disrupt the mitochondrial membrane efficiently, activating the Caspase-3 enzyme, and ultimately induces cell apoptosis. It has been observed from in vitro study that the degree of apoptosis for f -SAPNs was increased from 25.6% to 96.3%, while decreased degree of necrosis from 51.7% to 0.2% compared with its parent peptide analog (Cy5.5-c[RGDKLAK]; f -CP) occurs. Further investigations revealed that these f -SAPNs showed high uptake in U87MG glioblastoma cells in comparison with PC-3 prostate cancer cells. Moreover, in vivo therapeutic studies represented the prominent decrease in the size of tumor tissue treated with f -CP and f -SAPNs (201 ± 13 mm 3 and 104 ± 6 mm 3 , respectively) compared with untreated tumor tissues (366 ± 18 mm 3 ). These outcomes highlighted the specificity, and efficacy of f -SAPNs toward α v β 3 -integrin expressing tumor tissue in vivo and suggested that these novel designed f -SAPNs may serve as a potential theranostic drug for brain tumor glioblastoma multiforme. Graphical abstract The pH-sensitive method gives NIRF dye-coupled self-assembled peptide nanoparticle ( f -SAPNs), enables the tunable synthesis of spherical nanoparticles with high stability towards proteolysis, improved biocompatibility, and promising therapeutic efficacy.