RT Journal Article
SR Electronic
T1 bis-Cholesteryl-Conjugated Phosphorothioate Oligodeoxynucleotides Are Highly Selectively Taken Up by the Liver
JF Journal of Pharmacology and Experimental Therapeutics
JO J Pharmacol Exp Ther
FD American Society for Pharmacology and Experimental Therapeutics
SP 619
OP 626
DO 10.1124/jpet.302.2.619
VO 302
IS 2
A1 Martin K. Bijsterbosch
A1 Muthiah Manoharan
A1 Rick Dorland
A1 Richard van Veghel
A1 Erik A. L. Biessen
A1 Theo J. C. van Berkel
YR 2002
UL http://jpet.aspetjournals.org/content/302/2/619.abstract
AB We previously modulated, by conjugating a single cholesterol, plasma protein binding and liver cell uptake of a phosphorothioate oligodeoxynucleotide (PS-ODN). In this study, we investigated the biological fate of a PS-ODN, denoted ISIS-9389 (3′,5′-bis-cholesteryl-conjugated ISIS 3082), provided with two cholesteryl moieties. After intravenous injection of into rats, [3H]ISIS-9389 was cleared from plasma with a half-life of 23.6 ± 0.3 min. After 90 min (approximately 95% cleared), the liver contained 83.0 ± 0.8% of the dose. Spleen and bone (marrow), which constitute with the liver the reticuloendothelial system, contained 3.1 ± 0.3 and 4.3 ± 0.2%, respectively. All other tissues accumulated together <5% of the dose. The hepatic uptake of [3H]ISIS-9389 occurred mainly by endothelial cells (51.9 ± 6.4% of the liver uptake). Parenchymal and Kupffer cells were responsible for 24.9 ± 7.7 and 23.3 ± 2.5%, respectively. Preinjected polyinosinic acid and polyadenylic acid reduced hepatic uptake, albeit the latter was less effective. This finding suggests implication of (multiple) scavenger receptors in liver uptake of ISIS-9389. The interaction of ISIS-9389 with plasma proteins, analyzed by size exclusion chromatography, differs from that of unconjugated PS-ODN and PS-ODN with a single cholesterol. Plasma-incubated ISIS-9389 was mainly recovered as a high molecular weight complex. In conclusion, conjugation of PS-ODNs with two cholesteryl moieties results in almost quantitative uptake by the liver. The liver targeting exceeds the already impressive gain in liver uptake achieved by conjugation of a single cholesterol, and is expected to increase the therapeutic activity against liver-associated targets and reduce side effects in nonhepatic tissues. The American Society for Pharmacology and Experimental Therapeutics