| Description | Danoprevir (RG7227) is a peptidomimetic inhibitor of the NS3/4A protease(IC50=0.2-3.5 nM) of hepatitis C virus (HCV). |
| In vivo | 在大鼠或猴子中，给予Danoprevir（30 mg/kg）。 |
| Cell experiments | Serially diluted Danoprevir is added to Huh7 cells harboring the K2040 replicon 1 day after cell plating. For antiviral assays, after a 48-hour incubation, intracellular RNA is extracted, and the level of HCV replicon RNA is quantified by reverse transcription (RT)-PCR assay with the primers (5'-CACTCCCCTGTGAGGAACTACTG-3' and 5'-AGGCTGCACGACACTCATACT-3') and a probe (5'-6-FAM-CTTCACGCAGAAAGCGTCTAGCCATGG-MGBNFQ-3' using an ABI Prism 7900 sequence detection system. Here, FAM is 6-carboxyfluorescin and MGBNFQ is a molecular-groove binding non-fluorescence quencher specific to the HCV 5' untranslated region. Single-tube reactions are performed using the TaqMan Gold RT-PCR kit. Triplicate reactions for the RNA standards and samples are performed in 50 μL with 5 μL intracellular RNA (50 ng). RT is carried out at 48 °C for 30 min followed by 10 min at 95 °C. The PCR is run as follows: 15 seconds at 95 °C and 1 min at 60 °C for 40 cycles. Each RNA concentration is determined in triplicate. The absolute concentration of replicon RNA is calculated based on its signal relative to that of a standard curve generated by known concentrations of an in vitro-transcribed RNA corresponding to a genotype 1b 5' untranslated region. Replicon levels in the presence of Danoprevir are fitted to a four-parameter logistic function to obtain EC50.(Only for Reference) |
| Target activity | HCV NS3/4A protease:0.2 nM-3.5 nM |
| Synonyms | R7227, 丹诺普韦, ITMN-191, RG7227, RO5190591 |
| molecular weight | 731.83 |
| Molecular formula | C35H46FN5O9S |
| CAS | 850876-88-9 |
| Storage | Powder: -20°C for 3 years | In solvent: -80°C for 1 year |
| Solubility | H2O: < 1 mg/mL (insoluble or slightly soluble) DMSO: 133 mg/mL (181.7 mM) Ethanol: 133 mg/mL (181.7 mM) |
| References | 1. Seiwert SD, et al. Antimicrob Agents Chemother, 2008, 52(12), 4432-4441. 2. Bartels DJ, et al. J Infect Dis, 2008, 198(6), 800-807. 3. Imhof I, et al. Hepatology, 2011, 53(4), 1090-1099. |