Biological studies of an ICG-tagged aptamer as drug delivery system for malignant melanoma.

Malignant melanoma accounts for about 1% of all skin malignant tumors and represents the most aggressive and lethal form of skin cancer. Clinically, there exist different therapeutic options for melanoma treatment, such as surgery, chemotherapy, radiotherapy, photodynamic therapy and immunotherapy. However, serious adverse effects usually arise, and survival rates are still low because a high number of patients present relapses within 6-9 months after therapy. AS1411 is a G-quadruplex (G4) aptamer capable of tumor-specific recognition, since it binds to nucleolin, a multi-functional protein expressed in many different types of cancer cells. In this work, we present a novel drug delivery system composed of AS1411 and indocyanine green (ICG) to track its accumulation in tumoral cells in a melanoma mouse model. Using a simple supramolecular strategy, we conjugated the complex AS1411-ICG with C ligand, an acridine orange derivative with potential anticancer ligand. Then, we performed in vitro cytotoxicity experiments using the B16 mouse melanoma cell line, and in vivo experiments using a B16 mouse melanoma model to study biodistribution and histological changes. The circular dichroism (CD) data suggest that C does not affect the parallel G4 topology of AS1411-ICG, whereas it increases its thermal stability. Incubation of B16 melanoma cells with the AS1411-ICG complex associated with C increases the cytotoxicity compared with AS1411-ICG alone. From the in vivo studies, we conclude that both AS1411-ICG and AS1411-ICG-C presented the potential to accumulate preferentially in tumor tissues. Moreover, these compounds seem to be efficiently removed from the mice's bodies through kidney clearance. In summary, these results suggest that these complexes derived from AS1411 aptamer could act as a delivery system of ligands with antitumoral activity for in vivo melanoma therapy.