Asunaprevir inhibited HCV replicon reporter activity with genotype- and format-dependent potency. Stable genotype 1a and 1b replicons gave EC50 values of 4.0 ± 0.3 and 1.2 ± 0.3 nM; the stable genotype 3a NS3-hybrid replicon gave 1,162 ± 274 nM (means ± SD). These replicon results do not establish uniform activity across clinical isolates. [1]
In transporter-expressing HEK293 cells, asunaprevir inhibited OATP1B1-mediated E2G uptake and OATP1B3-mediated CCK-8 uptake, with IC50 values of 0.79 ± 0.21 and 0.65 ± 0.26 μM, respectively (means ± SD). The paper reports AdooQ-sourced material, without a catalog or lot identifier. These cell-assay findings alone do not establish a clinically significant drug interaction. [2]
Competitive NS3/4A inhibition and genotype dependence
Biochemical assay · Purified recombinant full-length HCV NS3/4A protease complexes representing genotypes 1–6 [1]
ASV competitively inhibited NS3/4A, with markedly weaker inhibition of genotype 3a than genotype 1 enzymes. VX-950 was the reference inhibitor; its 30-min preincubation differed from ASV, which did not require preincubation to enhance potency.
Literature-Reported Treatment Conditions
- Method
- FRET peptide-cleavage assay; substrate-kinetic analysis for competitive inhibition.
- Reported limits
- Full assay implementation is referenced to earlier methods; no operational reconstruction. IC50 and Ki are endpoints, not treatment doses.
Methods and source locations
Compound reported in the paper: Asunaprevir (BMS-650032); Compound label in the paper: ASV
Published compound-level evidence; no validation of the present A11295 lot or catalog solid-state form.
Results: Proteolytic enzyme activity; Table 1 and footnotes; Fig. 2; Methods: HCV NS3/4A protease genotype assays
NS3/4A protease activity, genotype 1a H77 and genotype 1b J4L6S: FRET; means ± SD from at least three independent experiments. — Table 1, genotype 1a and 1b rows and footnotes a–b
NS3/4A activity, genotype 3a S52: FRET; mean ± SD from at least three independent experiments. — Table 1, genotype 3a row and footnote a
Inhibition of genotype 1a and 1b NS3/4A protease: Kinetic Results report increasing apparent Km with unchanged Vmax; Fig. 2 is a representative experiment, not exact Ki read from the graph. — Results: Proteolytic enzyme activity, kinetic paragraph; Fig. 2A–B
Genotype-dependent activity in HCV replicon assays
Cell-based study · Stable and transient HCV replicon assays; genotypes 2b, 3a and 4a use NS3 protease hybrid-derived replicons [1]
ASV potency depended on genotype and assay format. Stable genotype 1a/1b replicons were more sensitive than stable genotype 2a and the genotype 3a hybrid. These are reporter-based cellular endpoints, not infection-cure measurements.
Literature-Reported Treatment Conditions
- Method
- Luciferase reporter endpoint in Table 3; VX-950 reference inhibitor. Methods describe DMSO comparison and paired viability assessment in replicon wells.
- Reported limits
- Exact replicon antiviral exposure duration is not specified in the selected endpoint paragraph; a 4-day cytotoxicity condition is not assigned to the antiviral EC50 experiment. Construction/culture/infection procedures and prior assay-method papers not extracted.
Methods and source locations
Compound reported in the paper: Asunaprevir (BMS-650032); Compound label in the paper: ASV
Published compound-level evidence; no validation of the present A11295 lot or catalog solid-state form.
Table 3 and footnotes a–c; Results: Antiviral activity in cell culture; Methods: selected reporter endpoint and cytotoxicity paragraphs
Reporter inhibition in stable replicons: Luciferase reporter; means ± SD from at least three independent experiments. VX-950 comparator values for these rows: 995 ± 11, 427 ± 3, 229 ± 32, and 2,731 ± 188 nM, respectively. — Table 3, four stable cell-line rows; footnotes a–c
Reporter inhibition in transient replicon assays: Luciferase reporter; means ± SD from at least three independent experiments. Assay formats remain explicitly separate within one coherent genotype-profiling study. — Table 3, four transient rows; footnotes a–c
Inhibition of OATP1B1 and OATP1B3 substrate uptake
Cell-based study · HEK293 cells expressing human OATP1B1 or OATP1B3, with mock-transfected HEK293 background subtraction [2]
ASV inhibited uptake through both human transporters in a cell assay. These substrate-specific IC50 values indicate interaction potential; they do not alone establish a clinically important drug interaction.
Literature-Reported Treatment Conditions
- Treatment concentration
- 0.01–10 μM ASV; concentration series, not IC50-as-treatment-dose.
- Exposure duration
- Uptake: 3 min for OATP1B1; 5 min for OATP1B3.
- Method
- Coincubation with E2G (100 nM) for OATP1B1 or CCK-8 (10 nM) for OATP1B3. Mock-cell uptake subtracted; DMSO-only transporter uptake set to 100%. Rifampin/BSP used in functional validation.
- Vehicle
- DMSO
- Reported limits
- No current lot validation; no human clinical experiment in this study.
Methods and source locations
Compound reported in the paper: Asunaprevir (BMS-650032); Compound label in the paper: ASV
Published compound-level evidence; no validation of the present A11295 lot or catalog solid-state form.
Methods: Transporter inhibition assays (coincubation method); Results: coincubation inhibition; Fig. 2; Table 2
Inhibition of OATP1B1-mediated E2G uptake: Mean ± SD, three separate experiments, each performed in duplicate; Fig. 2 caption defines replication and normalization. — Table 2, ASV/OATP1B1 IC50; Fig. 2 caption
Inhibition of OATP1B3-mediated CCK-8 uptake: Mean ± SD, three separate experiments, each performed in duplicate; Fig. 2 caption defines replication and normalization. — Table 2, ASV/OATP1B3 IC50; Fig. 2 caption
