Physicochemical and disposition characteristics of antisense oligonucleotides complexed with glycosylated...

The disposition characteristics of a 20 mer antisense phosphodiester oligonucleotide (PO) and its fully phosphorothioated derivative (PS) alone or complexed with glycosylated poly(L-lysine) (galactosylated polylysine, Gal-PLL; mannosylated polylysine, Man-PLL) were studied in mice in relation to their physicochemical characteristics. Good complex formation was obtained at a ratio of 1:0.6, w/w [oligonucleotides (ODNs)/carrier]. The 1:0.6 weight ratio of ODNs/Gal-PLL and ODNs/Man-PLL complexes had zeta potentials of -27 to -31 mV and mean particle size of 100 to 160 nm. After intravenous injection, 35S-labeled ODNs were eliminated rapidly from the circulation; however, their organ disposition characteristics depended on their type. Complex formation with glycosylated PLL increased the hepatic uptake and decreased the urinary clearance of these ODNs to a great extent. These complexes were taken up by both liver parenchymal cells (PC) and nonparenchymal cells (NPC). However, ODNs/Gal-PLL complexes showed a fairly high PC concentration, whereas ODNs/Man-PLL complexes distributed equally to both PC and NPC. The hepatic uptakes of PS/Gal-PLL and PS/Man-PLL complexes were partially inhibited by prior administration of Gal-BSA and Man-BSA, respectively, suggesting their hepatic uptake via the respective receptor-mediated endocytosis. However, uptake by galactose receptors of Kupffer cells, zeta potential, particle size, and Kupffer cell phagocytosis also seem to influence their uptake process. In conclusion, this study illustrates that ODNs can be delivered to hepatocytes and macrophages via galactose and mannose receptors, respectively.