Target‐Selective Drug Delivery through Liposomes Labeled with Oligobranched Neurotensin Peptides

The structure and the in vitro behavior of liposomes filled with the cytotoxic drug doxorubicin (Doxo) and functionalized on the external surface with a branched moiety containing four copies of the 8–13 neurotensin (NT) peptide is reported. The new functionalized liposomes, DOPC‐NT 4 Lys(C 18 ) 2 , are obtained by co‐aggregation of the DOPC phospholipid with a new synthetic amphiphilic molecule, NT 4 Lys(C 18 ) 2 , which contains a lysine scaffold derivatized with a lipophilic moiety and a tetrabranched hydrophilic peptide, NT8–13, a neurotensin peptide fragment well known for its ability to mimic the neurotensin peptide in receptor binding ability. Dynamic light scattering measurements indicate a value for the hydrodynamic radius (RH) of 88.3±4.4 nm. The selective internalization and cytotoxicity of DOPC‐NT 4 Lys(C 18 ) 2 liposomes containing Doxo, as compared to pure DOPC liposomes, were tested in HT29 human colon adenocarcinoma and TE671 human rhabdomyosarcoma cells, both of which express neurotensin receptors. Peptide‐functionalized liposomes show a clear advantage in comparison to pure DOPC liposomes with regard to drug internalization in both HT29 and TE671 tumor cells: FACS analysis indicates an increase in fluorescence signal of the NT 4 ‐liposomes, compared to the DOPC pure analogues, in both cell lines; cytotoxicity of DOPC‐NT 4 Lys(C 18 ) 2 ‐Doxo liposomes is increased four‐fold with respect to DOPC‐Doxo liposomes in both HT29 and TE671 cell lines. These effects could to be ascribed to the higher rate of internalization for DOPC‐NT 4 Lys(C 18 ) 2 ‐Doxo liposomes, due to stronger binding driven by a lower dissociation constant of the NT 4 ‐liposomes that bind the membrane onto a specific protein, in contrast to DOPC liposomes, which approach the plasma membrane unselectively.