TABLE 1

RAM frequencies in vitro and potency of EDP-239 against NS5A mutations in transient GT1 replicons

HCV genotypeWild type or mutant repliconaNo. (%) of clones with substitutionbEDP-239 EC50 (pM) (mean ± SD)cFold resistancedReplication efficiency (%)e
1a (H77c)Wild type3.6 ± 0.4100
K24G21 (25)177.4 ± 41.750179
K24R34 (41)10.8 ± 2.03135
K24T5 (6.0)5.1 ± 1.81121
M28T10 (12)1,191 ± 22233126
M28V6 (7.1)24.0 ± 3.3793
Q30R19 (23)1,938 ± 522538106
Q30H13 (15)137.3 ± 30.63869
Q30P2 (2.4)4.9 ± 2.81183
L31M3 (3.6)284 ± 6579114
F36L58 (69)4.4 ± 1.21173
Y93C2 (2.4)1,908 ± 6005306
Y93H6 (7.1)58,103 ± 25,59516,1401
Y93N2 (2.4)115,479 ± 40,07132,0782
K24R F36L2 (2.4)116.6 ± 48.73221
1b (Con1)Wild type2.7 ± 1.01100
L31M3 (4.7)5.6 ± 0.52240
L31V22 (34)54.6 ± 9.020338
A92V14 (22)3.2 ± 0.71200
Y93H42 (66)58.5 ± 7.12233
A92V Y93H10 (16)1,917 ± 23271040
L31V Y93H1 (1.6)172,012 ± 16,25363,70848
L31V A92V Y93H1 (1.6)476,379 ± 61,883176,43714
  • 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.