caa a22 4500 477127428 CHVBK 20180405111654.0 cr unu---uuuuu 170330e19970401xx s 000 0 eng 10.1023/A:1012116003706 doi (NATIONALLICENCE)springer-10.1023/A:1012116003706 Hepatic Distribution and Clearance of Antisense Oligonucleotides in the Isolated Perfused Rat Liver [Elektronische Daten] [Arno Nolting, Robert DeLong, Michael Fisher, Eric Wickstrom, Gary Pollack, Rudolph Juliano, Kim Brouwer] Purpose. This study was conducted to investigate the impact of backbone modifications on the hepatobiliary disposition of oligonucleotides. Methods. The disposition of backbone-modified antisense oligonucleotides [phosphorothioate (PS) and methylphosphonate (MP)] of the same base-length and sequence (5′-TAC-GCC-AAC-AGC-TCC-3′), complementary to the codon 12 activating mutation of Ki-ras, was investigated in the isolated perfused rat liver. Livers were perfused for 2 hr; perfusate and bile concentrations were analyzed by HPLC. Hepatocellular distribution was examined by measuring the amount of radiolabeled PS oligonucleotide associated with hepatocytes and Kupffer cells. Protein binding of the PS and MP oligonucleotides was determined in rat serum by ultrafiltration. Results. MP oligonucleotide perfusate concentrations remained constant during the 2-hour perfusion. In contrast, PS oligonucleotide was eliminated slowly by the isolated perfused liver [Cl = 1.05 ± 0.21 mL/min; extraction ratio = 0.06 ± 0.01]. Uptake of PS oligonucleotide by Kupffer cells appeared to exceed uptake by hepatocytes, based on standard cell separation techniques as well as confocal microscopy. The degree of protein binding in rat serum was greater for the PS oligonucleotide (79.9 ± 2.2%) than for the MP oligonucleotide (53.0 ± 4.7%). Conclusions. Backbone modifications significantly influence the hepatic clearance of oligonucleotides. Uncharged MP oligonucleotides are not extracted by the isolated perfused rat liver, whereas the charged PS oligonucleotide is processed more readily. Plenum Publishing Corporation, 1997 isolated perfused rat liver nationallicence oligonucleotides nationallicence hepatocellular distribution nationallicence Nolting Arno Division of Pharmaceutics, School of Pharmacy, University of North Carolina, 27599, Chapel Hill, North Carolina aut DeLong Robert Department of Pharmacology, School of Medicine, University of North Carolina, 27599, Chapel Hill, North Carolina aut Fisher Michael Department of Pharmacology, School of Medicine, University of North Carolina, 27599, Chapel Hill, North Carolina aut Wickstrom Eric Department of Pharmacology, Thomas Jefferson University, 19107, Philadelphia, Pennsylvania aut Pollack Gary Division of Pharmaceutics, School of Pharmacy, University of North Carolina, 27599, Chapel Hill, North Carolina aut Juliano Rudolph Department of Pharmacology, School of Medicine, University of North Carolina, 27599, Chapel Hill, North Carolina aut Brouwer Kim Division of Pharmaceutics, School of Pharmacy, University of North Carolina, 27599, Chapel Hill, North Carolina aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/4(1997-04-01), 516-521 0724-8741 14:4<516 1997 14 11095 https://doi.org/10.1023/A:1012116003706 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012116003706 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nolting Arno Division of Pharmaceutics, School of Pharmacy, University of North Carolina, 27599, Chapel Hill, North Carolina aut NATIONALLICENCE 700 1- DeLong Robert Department of Pharmacology, School of Medicine, University of North Carolina, 27599, Chapel Hill, North Carolina aut NATIONALLICENCE 700 1- Fisher Michael Department of Pharmacology, School of Medicine, University of North Carolina, 27599, Chapel Hill, North Carolina aut NATIONALLICENCE 700 1- Wickstrom Eric Department of Pharmacology, Thomas Jefferson University, 19107, Philadelphia, Pennsylvania aut NATIONALLICENCE 700 1- Pollack Gary Division of Pharmaceutics, School of Pharmacy, University of North Carolina, 27599, Chapel Hill, North Carolina aut NATIONALLICENCE 700 1- Juliano Rudolph Department of Pharmacology, School of Medicine, University of North Carolina, 27599, Chapel Hill, North Carolina aut NATIONALLICENCE 700 1- Brouwer Kim Division of Pharmaceutics, School of Pharmacy, University of North Carolina, 27599, Chapel Hill, North Carolina aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/4(1997-04-01), 516-521 0724-8741 14:4<516 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer