TABLE 1.
RAM frequencies in vitro and potency of EDP-239 against NS5A mutations in transient GT1 replicons
| HCV genotype | Wild type or mutant replicona | No. (%) of clones with substitutionb | EDP-239 EC50 (pM) (mean ± SD)c | Fold resistanced | Replication efficiency (%)e |
|---|---|---|---|---|---|
| 1a (H77c) | Wild type | 3.6 ± 0.4 | 100 | ||
| K24G | 21 (25) | 177.4 ± 41.7 | 50 | 179 | |
| K24R | 34 (41) | 10.8 ± 2.0 | 3 | 135 | |
| K24T | 5 (6.0) | 5.1 ± 1.8 | 1 | 121 | |
| M28T | 10 (12) | 1,191 ± 222 | 331 | 26 | |
| M28V | 6 (7.1) | 24.0 ± 3.3 | 7 | 93 | |
| Q30R | 19 (23) | 1,938 ± 522 | 538 | 106 | |
| Q30H | 13 (15) | 137.3 ± 30.6 | 38 | 69 | |
| Q30P | 2 (2.4) | 4.9 ± 2.8 | 1 | 183 | |
| L31M | 3 (3.6) | 284 ± 65 | 79 | 114 | |
| F36L | 58 (69) | 4.4 ± 1.2 | 1 | 173 | |
| Y93C | 2 (2.4) | 1,908 ± 600 | 530 | 6 | |
| Y93H | 6 (7.1) | 58,103 ± 25,595 | 16,140 | 1 | |
| Y93N | 2 (2.4) | 115,479 ± 40,071 | 32,078 | 2 | |
| K24R F36L | 2 (2.4) | 116.6 ± 48.7 | 32 | 21 | |
| 1b (Con1) | Wild type | 2.7 ± 1.0 | 1 | 100 | |
| L31M | 3 (4.7) | 5.6 ± 0.5 | 2 | 240 | |
| L31V | 22 (34) | 54.6 ± 9.0 | 20 | 338 | |
| A92V | 14 (22) | 3.2 ± 0.7 | 1 | 200 | |
| Y93H | 42 (66) | 58.5 ± 7.1 | 22 | 33 | |
| A92V Y93H | 10 (16) | 1,917 ± 232 | 710 | 40 | |
| L31V Y93H | 1 (1.6) | 172,012 ± 16,253 | 63,708 | 48 | |
| L31V A92V Y93H | 1 (1.6) | 476,379 ± 61,883 | 176,437 | 14 |
a
NS5A mutations selected by EDP-239 and cloned into transiently transfected GT1 replicons.
b
Number of clones and frequency at which mutations were identified out of the total number of colonies sequenced for genotypes 1a (84 total) and 1b (64 total).
c
Averages for at least 6 biological replicates.
d
Calculated from the ratio of the EC50s for the mutant to that of the wild-type replicon.
e
Calculated as a percentage of mutant replicon replication relative to replication of the wild-type replicon.