Gallic Acid–Triethylene Glycol Aptadendrimers Synthesis, Biophysical Characterization and Cellular Evaluation

André Miranda; Roi Lopez-Blanco; Jéssica Lopes-Nunes; Ana M. Melo; Maria Paula Cabral Campello; António Paulo; Maria Cristina Oliveira; et al

Informações chave

Autores:

André Miranda; Roi Lopez-Blanco; Jéssica Lopes-Nunes; Ana M. Melo; Maria Paula Cabral Campello (Maria Paula Cordeiro Crespo Cabral Campello Aboim de Barros); António Paulo (António Manuel Rocha Paulo); Maria Cristina Oliveira (Maria Cristina das Neves Oliveira); et al

Publicado em

14/11/2022

Resumo

Herein, we describe the synthesis of an aptadendrimer by covalent bioconjugation of a gallic acid–triethylene glycol (GATG) dendrimer with the G-quadruplex (G4) AT11 aptamer (a modified version of AS1411) at the surface. We evaluated the loading and interaction of an acridine orange ligand, termed C8, that acts as an anticancer drug and binder/stabilizer of the G4 structure of AT11. Dynamic light scattering experiments demonstrated that the aptadendrimer was approximately 3.1 nm in diameter. Both steady-state and time-resolved fluorescence anisotropy evidenced the interaction between the aptadendrimer and C8. Additionally, we demonstrated that the iodine atom of the C8 ligand acts as an effective intramolecular quencher in solution, while upon complexation with the aptadendrimer, it adopts a more extended conformation. Docking studies support this conclusion. Release experiments show a delivery of C8 after 4 h. The aptadendrimers tend to localize in the cytoplasm of various cell lines studied as demonstrated by confocal microscopy. The internalization of the aptadendrimers is not nucleolin-mediated or by passive diffusion, but via endocytosis. MTT studies with prostate cancer cells and non-malignant cells evidenced high cytotoxicity mainly due to the C8 ligand. The rapid internalization of the aptadendrimers and the fluorescence properties make them attractive for the development of potential nanocarriers.