Cell experiment:

For the determination of cytotoxicity of JTK-853, the Huh-7.5 cells are treated with JTK-853 for 2 weeks. The thumb pocket NNI-B and NS5Ai are added at 100 μM and 100 nM, respectively. JTK-853 is added at 10 μM. JTK-853-containing medium is changed twice a week. Two weeks after the culture, the cells are stained with crystal violet [1% (v/v) in methanol], and then lysed by the lysis buffer. The cytotoxicity is determined as a measurement of OD 595 nm of the cell lysates[1].

JTK-853 is a novel, non-nucleoside Hepatitis C Virus (HCV) polymerase inhibitor which shows effective antiviral activity in HCV replicon cells with EC50s of 0.38 and 0.035 ?M in genotype 1a H77 and 1b Con1 strains, respectively.

JTK-853 is a novel, non-nucleoside Hepatitis C Virus Polymerase inhibitor which shows effective antiviral activity in HCV replicon cells with EC50s of 0.38 and 0.035 ?M in genotype 1a H77 and 1b Con1 strains, respectively. When JTK-853 is incubated with the replicon cells for 48 h, it shows antiviral activity against genotype 1a H77 and 1b Con1 replicon cells with EC90 values of 6.5±0.5 and 0.34±0.05 ?M, respectively. At 10 ?M, JTK-853 induces apparent Huh-7.5 cell death in 2-week culture. JTK-853 suppresses the drug-resistant colony formation in the genotype 1a replicon cells, and the numbers of JTK-853-resistant colonies are much lower than those of GS-9190-resistant colonies for both genotypes[1].

[1]. Ando I, et al. JTK-853, a novel non-nucleoside hepatitis C virus polymerase inhibitor, demonstrates a high genetic barrier to resistance in vitro. Intervirology. 2013;56(5):302-9.