Resistance Analysis of the Hepatitis C Virus NS5A Inhibitor BMS-790052 in an In Vitro Replicon System

Antimicrobial Agents and Chemotherapy, September 2010, p. 3641-3650, Vol. 54, No. 9
0066-4804/10/$12.00+0 doi:10.1128/AAC.00556-10
Copyright © 2010, American Society for Microbiology. All Rights Reserved.

Robert A. Fridell,* Dike Qiu, Chunfu Wang, Lourdes Valera, and Min Gao

Department of Virology, Bristol-Myers Squibb Research and Development, Wallingford, Connecticut

Received 23 April 2010/ Returned for modification 27 May 2010/ Accepted 23 June 2010

BMS-790052 is the most potent hepatitis C virus (HCV) inhibitor reported to date, with 50% effective concentrations (EC50s) of 50 pM against genotype 1 replicons. This exceptional potency translated to rapid viral load declines in a phase I clinical study. By targeting NS5A, BMS-790052 is distinct from most HCV inhibitors in clinical evaluation. As an initial step toward correlating in vitro and in vivo resistances, multiple cell lines and selective pressures were used to identify BMS-790052-resistant variants in genotype 1 replicons. Similarities and differences were observed between genotypes 1a and 1b. For genotype 1b, L31F/V, P32L, and Y93H/N were identified as primary resistance mutations. L23F, R30Q, and P58S acted as secondary resistance substitutions, enhancing the resistance of primary mutations but themselves not conferring resistance. For genotype 1a, more sites of resistance were identified, and substitutions at these sites (M28T, Q30E/H/R, L31M/V, P32L, and Y93C/H/N) conferred higher levels of resistance. For both subtypes, combining two resistance mutations markedly decreased inhibitor susceptibility. Selection studies with a 1b/1a hybrid replicon highlighted the importance of the NS5A N-terminal region in determining genotype-specific inhibitor responses. As single mutations, Q30E and Y93N in genotype 1a conferred the highest levels of resistance. For genotype 1b, BMS-790052 retained subnanomolar potency against all variants with single amino acid substitutions, suggesting that multiple mutations will likely be required for significant in vivo resistance in this genetic background. Importantly, BMS-790052-resistant variants remained fully sensitive to alpha interferon and small-molecule inhibitors of HCV protease and polymerase.

* Corresponding author. Mailing address: Department of Virology, Bristol-Myers Squibb Research and Development, 5 Research Parkway, Wallingford, CT 06492. Phone: (203) 677-7034. Fax: (203) 677-6088. E-mail: robert.fridell@bms.com

Published ahead of print on 28 June 2010.

Present address: Discovery Biology, Bristol-Myers Squibb Research and Development, Hopewell, NJ.

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