Table 4.

Characterization of gt 1a variants identified in the dual selections using transient replication assays

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

^aReplicons 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.

^bFold 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.

^c325, BMS-791325.