Table 4.
Selection condition | Varianta |
Fold resistanceb |
Replication efficiency relative to WT level | ||||
---|---|---|---|---|---|---|---|
NS3 | NS5A | NS5B | Asunaprevir (ASV) | Daclatasvir (DCV) | 325c | ||
10× or 30× ASV | R155K | — | — | 17–48 | 1 | 1 | 0.3–0.5 |
10× or 30× DCV | — | Q30H-K68R | — | 1 | 1,280–2,000 | 3 | 0.04–0.07 |
10× or 30× 325 | — | — | A421V | 1 | 1 | 1–3 | 0.3–0.9 |
— | — | P495L | 2 | 1 | 88–100 | 0.07–0.1 | |
30× DCV/ASV | R155K | Q30H-K68R | — | 46–68 | 667–700 | 1 | 0.5–0.8 |
R155K | M28T-K68R | — | 34–61 | 1,125–13,333 | 1 | 0.4–1 | |
D168G | Q30H-K68R | — | 18–30 | 111–333 | 1 | 0.3–0.7 | |
30× DCV/325 | — | Q30H-K68R | A421V | 1 | 1000 | 2 | 0.8–1 |
— | Q30H-K68R | P495L | 2 | 1600 | 61–84 | 0.03–0.07 | |
— | Q30H-K68R | L392I | 2 | 2200 | 6–16 | 0.06–0.19 | |
Site-directed mutant | D168G | — | — | 8–16 | 1 | 1 | 0.07–0.09 |
Site-directed mutant | — | M28T | — | 1 | 750–820 | 1 | 0.2–0.4 |
Site-directed mutant | — | Q30H | — | 1 | 1111–1967 | 1 | 0.4–0.6 |
Site-directed mutant | — | K68R | — | 1 | 1 | 1 | 1–4.6 |
Site-directed mutant | — | — | L392I | 1 | 1 | 5–7 | 0.1–0.4 |
Replicons containing specific substitution(s) identified from selections were tested for resistance and replication fitness. Major selected variant constructs are in bold font. Site-directed mutants were not selected but were generated as controls in the WT backbone to assess impact on phenotype. For each selection condition, each row represents a unique variant that includes any linked mutations. —, WT sequence.
Fold resistance was determined by dividing the mutant HCV replicon EC50 by the WT replicon EC50. WT EC50s were 0.7 ± 0.3, 0.006 ± 0.002, and 2.2 ± 0.9 nM for ASV, DCV, and BMS-791325, respectively. Results represent the range of values determined from two or three independent experiments.
325, BMS-791325.