In Huh7 cells carrying a genotype 1b subgenomic replicon, ITMN-191 reduced intracellular replicon RNA with a reported 48-h EC50 of 1.8 nM. This cell-based potency endpoint is distinct from a treatment dose. [1]
Danoprevir also served as an input to an engineered transcription switch in HEK293T cells. It induced luciferase output, while red light partially suppressed that response; this reporter result does not measure antiviral activity. [2]
Preequilibrium inhibition of purified HCV NS3/4A
Biochemical assay · Full-length NS3/4A from K2040 genotype 1b and clinical-isolate genotype sequences [1]
ITMN-191 inhibited substrate cleavage. Genotype 3a was less susceptible than the K2040 reference under the same assay conditions. Initial-rate IC50 values do not describe full binding equilibrium.
Literature-Reported Treatment Conditions
- Exposure duration
- Initial-rate analysis up to 1 h
- Method
- FRET substrate cleavage; four-parameter logistic dose-response fit; no-inhibitor and no-enzyme controls.
- Reported limits
- Dose-series limits and inhibitor vehicle not specified in the selected biochemical method.
Methods and source locations
Compound reported in the paper: Danoprevir (ITMN-191/RG7227)
Evidence applies to the named experimental material and model; salt/form and current Adooq lot equivalence are not independently established.
Table 1 and footnote; Results: Biochemical inhibition of NS3/4A; Methods: Biochemical assays; Fig 2
Inhibition of K2040 genotype 1b NS3/4A substrate cleavage: Mean ± SD; minimum three independent experiments; preequilibrium initial-rate assay. — Table 1, K2040 row and footnote a; Methods: Biochemical assays
Inhibition of genotype 3a NS3/4A substrate cleavage: Mean ± SD; minimum three independent experiments; same preequilibrium assay. — Table 1, genotype 3a row and footnote a
Short-term genotype 1b replicon RNA reduction
Cell-based study · Huh7 cells harboring patient-derived genotype 1b K2040 subgenomic replicon [1]
A 48-h concentration-response experiment yielded a reported EC50 of 1.8 nM for intracellular replicon RNA reduction. This is a cell-based assay endpoint, not an administered treatment dose or an animal efficacy measurement.
Literature-Reported Treatment Conditions
- Treatment concentration
- 5 pM–100 nM
- Exposure duration
- 48 h
- Method
- Intracellular RNA quantified by RT-PCR; four-parameter logistic fit.
- Reported limits
- Vehicle and its final concentration for this 48-h assay are not specified in the read method; 0.1% DMSO from the separate clearance assay is not borrowed.
Methods and source locations
Compound reported in the paper: Danoprevir (ITMN-191/RG7227)
Evidence applies to the named experimental material and model; salt/form and current Adooq lot equivalence are not independently established.
Results: Cellular potency against an HCV subgenomic replicon, first paragraph; Fig 4A and caption; Methods: Cell-based assays, 48-h endpoint only
Reduction in intracellular genotype 1b replicon RNA: Reported fitted value in Results; Fig 4A gives mean percent reduction ± SD from 18 independent experiments. No uncertainty for the fitted EC50 is supplied there. — Results: Cellular potency, first paragraph; Fig 4A/caption
Danoprevir-responsive transcription with red-light suppression
Cell-based study · HEK293T cells carrying DNCR2-Gal4, NS3a-FHL and ΔPhyA-2×ZIM3 (PTRC_CIP-DNV) [2]
Danoprevir induced luciferase output in the engineered transcription system; red light reduced the induced output, with residual activity evident. This is an engineered reporter response, not HCV replication inhibition.
Literature-Reported Treatment Conditions
- Treatment concentration
- 10 μM danoprevir (1× working dilution of the reported 100×, 1 mM stock)
- Exposure duration
- 24 h light/dark comparison
- Method
- Luciferase readout; 660-nm red light at 1 mW/cm² or darkness.
- Vehicle
- Danoprevir stock in DMSO
- Reported limits
- Final vehicle matching and exact raw output values not established; SI/source-data archive not retrieved.
Methods and source locations
Compound reported in the paper: Danoprevir (ITMN-191/RG7227)
Evidence applies to the named experimental material and model; salt/form and current Adooq lot equivalence are not independently established.
Results: Small molecule- and red light-induced transcriptional regulation in mice, first two paragraphs; Fig 5c/caption; Methods: Small molecule induction, Light illumination, Luciferase reporter assay
Luciferase output from PTRC_CIP-DNV: Fig 5c: means ± SD; n=3 independent experiments, three technical replicates per experiment. No exact output or fold change estimated from bars. — Fig 5c and caption; Methods: Luciferase reporter assay and Statistical analysis
