Louis Sandra, Huybrecht T’jollyn, Nele Goeyvaerts, Xavier Woot De Trixhe, Jan Snoeys, Juan-Jose Perez-Ruixo, Anne-Gaëlle Dosne
Affiliation: Janssen Research & Development, Beerse , Belgium
Objectives: Silencing RNA (siRNA) is a novel therapeutic modality allowing for the specific downregulation of protein expression [1]. JNJ-73763989 (subsequently referred to as JNJ-3989), a N-acetylgalactosamine (GalNAc) siRNA combination product consisting of 2 triggers, is currently being developed as a treatment for chronic hepatitis B infection (HBV). GalNac-siRNAs show affinity for the asialoglycoprotein receptor (ASGPR), present on the plasma membrane of the hepatocyte, and allow efficient but saturable liver internalization via clathrin-mediated endocytosis [2]. The objective of this work was to describe the plasma and liver pharmacokinetics (PK) of JNJ-3989 in recombinant adeno-associated virus HBV (rAAV-HBV) infected mice while accounting for ASGPR-mediated liver internalization.
Methods: The analyzed data originated from a preclinical in-vivo experiment including 42 rAAV-HBV infected mice [3]. After infection, mice were monitored until stable viral parameters were achieved. A single dose of JNJ-3989 was administered either intravenous (IV) or subcutaneous (SC). Three SC dose levels (1, 3 and 10 mg kg-1) and one IV dose level (10 mg kg-1) were tested. Plasma PK samples were collected at 0.25, 0.75, 2, 4 hours after IV dose and 0.5, 1, 2, 4 hours after SC dose; an additional plasma sample was collected at sacrifice, i.e. at 1, 7, 21 and 42 days after dose depending on the cohort. Organs were harvested at sacrifice, including the liver.
A target-mediated drug disposition (TMDD) model with quasi-steady-state (QSS) approximations and peripheral compartment, was implemented to describe the free plasma concentrations and non-linear ASGPR-mediated liver internalization [4]. The TMDD QSS model was parameterized so that the total receptor concentration remained constant over time, as receptor recycling is expected to be very rapid (approx. between 10-15 min in human hepatocytes [5]) and the very limited plasma residence time of the drug (< 1 day) does not allow potential changes in protein expression to affect total receptor concentration. Receptor concentration was fixed in the model based on murine hepatic total ASGPR concentrations published by Bon et al. [6]. Liver concentrations expressed in nmol g-1 were scaled to nmol mL-1 based on the calculated relative hepatocyte density [7-8]. Additionally, volumes and clearances were scaled via fixed allometry centered around the sample mean total body weight of 26.7 g. The triggers were initially analyzed separately before being combined in a joint model where selected parameters were merged. Model building was performed based on improvement in objective function value (OFV), goodness-of-fit evaluations, acceptable parameter uncertainty, visual predictive checks and parsimony. All analyses were performed using NONMEM version 7.4 (using the FOCE algorithm) and R version 3.6.2.
Results: A total of 178 plasma and 84 liver PK observations from both triggers were included in the population PK analysis. The model provided an adequate fit to the plasma and liver PK data, with residual variability between 25 and 40% and relative standard errors below 20%. No inter-individual variability was found to be statistically significant. Plasma and liver elimination rates were found to be trigger-dependent. Both triggers distributed rapidly from plasma to liver, and showed a prolonged liver residence time (trigger S plasma t1/2: 0.17 hours versus hepatic t1/2: 11.1 days). The first-order internalization rate (kint) t1/2 was estimated to be 0.41 h, which is comparable with literature [9]. The proportion of the dose entering the liver was predicted to be 68%, 67% and 60% for SC doses of 1, 3 and 10 mg kg-1 respectively, while for 10 mg kg-1 IV this percentage dropped to 22%. This dose and route dependency likely reflected ASGPR saturation, which limits liver uptake at increased plasma concentrations observable after 10 mg kg-1 IV administration.
Conclusions: The TMDD QSS model provides a physiologically relevant description of the plasma and liver pharmacokinetics of JNJ-73763989 in AAV-HBV mice. This model may be used to link with PD endpoints measured in the same study.
References:
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Reference: PAGE 29 (2021) Abstr 9775 [www.page-meeting.org/?abstract=9775]
Poster: Drug/Disease Modelling - Infection