Background and Aims: Nucleotide analogue hepatitis C virus (HCV) polymerase inhibitors are considered a key component of directacting antiviral (DAA) combination therapies for chronic HCV infection treatment with pan-genotypic activity and high resistance barrier. JNJ-54257099 (JNJ-099) is a novel liver-targeted uridinebased nucleotide prodrug. Methods: The inhibitory activity of the nucleoside triphosphate (JNJ099 TP) against a recombinant HCV genotype (GT)1b NS5B polymerase was tested. JNJ-099 is not activated in HuH-7 cells; hence a phosphoramidate prodrug yielding the same TP as JNJ-099 was used for evaluating antiviral activity against HCV replicons harbouring NS5B sequences from multiple GTs or site-directed mutations associated with in vitro HCV inhibitor resistance. JNJ-099 TP formation in primary human hepatocytes (PHH) and cytotoxicity using a panel of human cell lines were also assessed. HCV GT1a- and GT3a-infected chimeric mice (pre-treatment serum HCV RNA level >106 copies/mL) with >70% human hepatocytes were treated with JNJ-099 for 7 days to assess anti-HCV activity in vivo. Results: JNJ-099 TP is a potent inhibitor of the HCV NS5B polymerase (IC50 1.0 μM, Ki 0.13 μM). A prodrug yielding the same active TP as JNJ-099 exhibited a broad HCV GT coverage with fold change in EC50 values compared with wild-type GT1b (FC) <2 for GT1a/b, GT2a/b, GT3a, GT4a and <3 for GT6a. Activity is retained against replicons harbouring mutations with in vitro resistance against NS5B inhibitors, including S282T (FC = 2.3). JNJ-099 TP was not a substrate for human mitochondrial RNA polymerase and demonstrated no inhibition of human DNA or RNA polymerases. No in vitro cytotoxicity was observed in a panel of human cell lines or in a high-content cytotoxicity screening assay up to the maximum concentrations tested. Incubation with JNJ-099 (10 μM) in PHH resulted in high concentrations of its TP from 2 h (87 pmol/106 PHH) with a max at 8h (172 pmol/106 PHH). Dose-dependent JNJ-099 anti-HCV activity was observed in HCV-infected chimeric mice with a median change from baseline at Day 7 of, respectively, −1.18 (GT1a) and −1.08 (GT3a) log10 HCV RNA copies/mL treated with 100 mg/kg/bid and -1.74 (GT3a) log10 HCV RNA copies/mL treated with 200 mg/kg/bid. Conclusions: JNJ-099, a potent and pan-genotypic uridine-based nucleotide analogue with a favourable resistance profile, is currently tested in Phase I as a potential novel component for chronic hepatitis C infection treatment.
In the ERA of New Direct Acting Antiviral Agents HCV Sequencing Allows the Most Accurate Subtype and Genotype Assignment / Aragri, M., Di Maio, V.C., Di Paolo, D., Cento, V., De Leonardis, F., Gianserra, L., Tontodonati, M., Micheli, V., Landonio, S., Manunta, A., Bertoli, A., Ciotti, M., Antonucci, F.P., Lenci, I., Francioso, S., Nicolini, L.A., Marenco, S., Teti, E., Lambiase, L., Milana, M., et al.. - In: JOURNAL OF HEPATOLOGY. - ISSN 0168-8278. - STAMPA. - 64:(2016), pp. S419-S420. (EASL International Liver Congress Barcelona, SPAIN APR 13-17, 2016) [10.1016/S0168-8278(16)00676-0].
In the ERA of New Direct Acting Antiviral Agents HCV Sequencing Allows the Most Accurate Subtype and Genotype Assignment
D’Ettorre, G.;Lichtner, M.;Mastroianni, C.;Taliani, G.;Pasquazzi, C.;
2016
Abstract
Background and Aims: Nucleotide analogue hepatitis C virus (HCV) polymerase inhibitors are considered a key component of directacting antiviral (DAA) combination therapies for chronic HCV infection treatment with pan-genotypic activity and high resistance barrier. JNJ-54257099 (JNJ-099) is a novel liver-targeted uridinebased nucleotide prodrug. Methods: The inhibitory activity of the nucleoside triphosphate (JNJ099 TP) against a recombinant HCV genotype (GT)1b NS5B polymerase was tested. JNJ-099 is not activated in HuH-7 cells; hence a phosphoramidate prodrug yielding the same TP as JNJ-099 was used for evaluating antiviral activity against HCV replicons harbouring NS5B sequences from multiple GTs or site-directed mutations associated with in vitro HCV inhibitor resistance. JNJ-099 TP formation in primary human hepatocytes (PHH) and cytotoxicity using a panel of human cell lines were also assessed. HCV GT1a- and GT3a-infected chimeric mice (pre-treatment serum HCV RNA level >106 copies/mL) with >70% human hepatocytes were treated with JNJ-099 for 7 days to assess anti-HCV activity in vivo. Results: JNJ-099 TP is a potent inhibitor of the HCV NS5B polymerase (IC50 1.0 μM, Ki 0.13 μM). A prodrug yielding the same active TP as JNJ-099 exhibited a broad HCV GT coverage with fold change in EC50 values compared with wild-type GT1b (FC) <2 for GT1a/b, GT2a/b, GT3a, GT4a and <3 for GT6a. Activity is retained against replicons harbouring mutations with in vitro resistance against NS5B inhibitors, including S282T (FC = 2.3). JNJ-099 TP was not a substrate for human mitochondrial RNA polymerase and demonstrated no inhibition of human DNA or RNA polymerases. No in vitro cytotoxicity was observed in a panel of human cell lines or in a high-content cytotoxicity screening assay up to the maximum concentrations tested. Incubation with JNJ-099 (10 μM) in PHH resulted in high concentrations of its TP from 2 h (87 pmol/106 PHH) with a max at 8h (172 pmol/106 PHH). Dose-dependent JNJ-099 anti-HCV activity was observed in HCV-infected chimeric mice with a median change from baseline at Day 7 of, respectively, −1.18 (GT1a) and −1.08 (GT3a) log10 HCV RNA copies/mL treated with 100 mg/kg/bid and -1.74 (GT3a) log10 HCV RNA copies/mL treated with 200 mg/kg/bid. Conclusions: JNJ-099, a potent and pan-genotypic uridine-based nucleotide analogue with a favourable resistance profile, is currently tested in Phase I as a potential novel component for chronic hepatitis C infection treatment.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
