Pharmacokinetics and tissue distribution of a ribozyme directed against hepatitis C virus RNA following subcutaneous or intravenous administration in mice.
A nuclease resistant ribozyme targeting the 5' untranslated region (5' UTR) of hepatitis C virus (HCV) at site 195 has been identified. To investigate the therapeutic utility of this ribozyme, we evaluated the pharmacokinetics and tissue distribution with two labeled forms of this ribozyme. [(32)P]-labeled ribozyme was administered as a single subcutaneous (SC) or intravenous (IV) bolus at a dose of 10 mg/kg or 30 mg/kg in C57Bl/6 mice. Regardless of route of administration, peak liver concentrations achieved were greater than the concentration necessary to inhibit HCV-IRES-luciferase expression in cell culture. The ribozyme was well absorbed after SC administration (89%) and had an elimination half-life of 23 minutes. To show intracellular localization of the ribozyme in target tissue, a tetramethyl rhodamine (TMR)-labeled ribozyme was administered as a single SC or IV bolus at a dose of 30 mg/kg in C57Bl/6 mice. Mice treated SC or IV with TMR-labeled ribozyme had positive fluorescence in the liver from 15 minutes to 48 hours after dosing. Definite positive fluorescence was still present at 72 hours in the mice dosed via the IV route. At early time points (15 and 30 minutes postinjection), nuclear and possibly cytoplasmic fluorescence was present in the hepatocytes, and sinusoidal fluorescence was intense. At the later time points, fluorescence became more punctate. Abundant staining was often present in Kupffer cells. This study confirms the retention of ribozyme in liver cells and supports the potential of an anti-HCV ribozyme as a therapeutic agent for treatment of chronic hepatitis C.
Lee
P A
PA
Ribozyme Pharmaceuticals Inc., Boulder, CO 80301, USA.
Blatt
L M
LM
Blanchard
K S
KS
Bouhana
K S
KS
Pavco
P A
PA
Bellon
L
L
Sandberg
J A
JA
D000818:Animals; D001483:Base Sequence / Q000235:genetics; D004279:DNA, Viral / Q000187:drug effects*; D005260:Female; D005456:Fluorescent Dyes; D016174:Hepacivirus / Q000235:genetics*; D007275:Injections, Intravenous; D007279:Injections, Subcutaneous; D007425:Intracellular Membranes / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009994:Osmolar Concentration; D010761:Phosphorus Radioisotopes; D016337:RNA, Catalytic / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000493:pharmacokinetics*; D012235:Rhodamines; D014018:Tissue Distribution
D004279:DNA, Viral; D005456:Fluorescent Dyes; D010761:Phosphorus Radioisotopes; D016337:RNA, Catalytic; D012235:Rhodamines; C005358:tetramethylrhodamine