[Control of in vivo characteristics of gene and antisense DNA disposition]

In order to construct a strategy for control of in vivo disposition characteristics of gene and antisense DNA, in vivo stability and basic pharmacokinetic properties of DNA were investigated. A model gene, plasmid DNA (pCAT), and model oligonucleotide (thymidine 10-mer; T10) derivatives with phosphoroamidate substitution at the 3' and/or 5'-terminal internucleotide linkage, were rapidly degraded in vivo after intravenous injection into mice. The degradation rates were much faster than those observed in in vitro experiments using plasma and whole blood, suggesting that they underwent enzymatic degradation in other compartments than the blood pool. More than 70% of injected pCAT was taken up by the liver within 5 minutes, and the uptake clearance was almost identical to the plasma flow rate in the liver. On the other hand, T10 derivatives were rapidly excreted in the urine and taken up by the kidney and liver. The urinary excretion clearance was close to the glomerular filtration rate. In an attempt to control T10 disposition characteristics, the oligonucleotide was conjugated to a macromolecular carrier, carboxymethyl dextran (CMD). The T10-CMD conjugate exhibited increased in vivo stability and prolonged plasma circulation time. Thus, the present study has shown that macromolecular conjugation is a useful approach to improve in vivo disposition of antisense oligonucleotides.