Aptamer-functionalized pH-sensitive liposomes for a selective delivery of echinomycin into cancer cells

Echinomycin (quinomycin A) is a peptide antibiotic from the quinoxaline family, which has a DNA bifunctional intercalating activity and an inhibitor of hypoxia-inducible factor (HIF1α). Echinomycin was discovered in 1957 as a potent antitumor agent; however, it was not successful in clinical use due to its low water solubility and short half-life. To revitalize this potent drug, it is important to increase its aqueous solubility and bioavailability. In this study, echinomycin was loaded into PEGylated pH-sensitive liposomes ( PEG Lip pH ) and functionalized with anti-nucleolin aptamer (Apt NCL ) for selective targeting and pH-responsive release of echinomycin into cancer cells. Echinomycin was complexed with γ-cyclodextrin (ECγCD) to enhance its water solubility and then encapsulated into pH-sensitive liposomes ( PEG Lip pH -ECγCD). Then, liposomes were functionalized with Apt NCL (Apt NCL-PEG Lip pH -ECγCD) and the successful functionalization was confirmed by dynamic light scattering (DLS) measurements and gel electrophoresis. Cellular uptake for Apt NCL-PEG Lip pH was evaluated by flow cytometry analysis using MDA-MB-231, MCF7, A549 cancer cell lines with respect to the normal fibroblast cells. The results showed a higher uptake and selectivity for Apt NCL-PEG Lip pH compared to PEG Lip pH . The anti-proliferative effects of Apt NCL-PEG Lip pH -ECγCD were more potent than PEG Lip pH -ECγCD by 3.5, 4, and 5 folds for A549, MDA-MB-231, and MCF7, respectively. Selectivity indices (SI) for Apt NCL-PEG Lip pH -ECγCD for the tumor cell lines compared to the normal cell line after 72 h were MDA-MB-231 (43.3), MCF7 (16.9), and A549 (8.5). Furthermore, SI after 3 h for the three cancer cell lines were 4.7, 2.5, 2.8, respectively. Echinomycin was loaded into PEGylated pH-sensitive liposomes and functionalized with anti-nucleolin aptamer for selective targeting and pH-responsive release of echinomycin into cancer cells.