Chemical genetics strategy identifies an HCV NS5A inhibitor with a potent clinical effect

Journal name:
Nature
Volume:
465,
Pages:
96–100
Date published:
DOI:
doi:10.1038/nature08960
Received
Accepted
Published online

The worldwide prevalence of chronic hepatitis C virus (HCV) infection is estimated to be approaching 200million people1. Current therapy relies upon a combination of pegylated interferon-α and ribavirin, a poorly tolerated regimen typically associated with less than 50% sustained virological response rate in those infected with genotype 1 virus2, 3. The development of direct-acting antiviral agents to treat HCV has focused predominantly on inhibitors of the viral enzymes NS3 protease and the RNA-dependent RNA polymerase NS5B4. Here we describe the profile of BMS-790052, a small molecule inhibitor of the HCV NS5A protein that exhibits picomolar half-maximum effective concentrations (EC50) towards replicons expressing a broad range of HCV genotypes and the JFH-1 genotype 2a infectious virus in cell culture. In a phase I clinical trial in patients chronically infected with HCV, administration of a single 100-mg dose of BMS-790052 was associated with a 3.3log10 reduction in mean viral load measured 24h post-dose that was sustained for an additional 120h in two patients infected with genotype 1b virus. Genotypic analysis of samples taken at baseline, 24 and 144h post-dose revealed that the major HCV variants observed had substitutions at amino-acid positions identified using the in vitro replicon system. These results provide the first clinical validation of an inhibitor of HCV NS5A, a protein with no known enzymatic function, as an approach to the suppression of virus replication that offers potential as part of a therapeutic regimen based on combinations of HCV inhibitors.

At a glance

Figures

  1. Structures of the iminothiazolidinone BMS-858, BMS-790052, the biotin-tagged HCV NS5A inhibitor 1 and its inactive stereoisomer 2.
    Figure 1: Structures of the iminothiazolidinone BMS-858, BMS-790052, the biotin-tagged HCV NS5A inhibitor 1 and its inactive stereoisomer 2.
  2. The active inhibitor 1 binds to genotype 1b HCV NS5A whereas the inactive stereoisomer 2 does not.
    Figure 2: The active inhibitor 1 binds to genotype 1b HCV NS5A whereas the inactive stereoisomer 2 does not.

    Genotype 1b replicon cells were exposed to either 1 or 2 for approximately 18h before the cells were lysed. A portion of the supernatant was set aside to serve as input control for immunoblot analysis (lanes 1 and 2) while the remainder was mixed with streptavidin–agarose beads and incubated for several hours. Bound proteins were detected by immunoblotting with primary antibodies directed to HCV NS5A. Lanes 1 and 3 depict the results of experiments with the active inhibitor compound 1 whereas lanes 2 and 4 depict the results with the inactive stereoisomer 2.

  3. Mean plasma concentration–time profile (time 0–72[thinsp]h) of BMS-790052 after single oral administration of 1–200[thinsp]mg of drug to healthy subjects.
    Figure 3: Mean plasma concentration–time profile (time 0–72h) of BMS-790052 after single oral administration of 1–200mg of drug to healthy subjects.

    In a double-blind, placebo-controlled, sequential, single ascending-dose study, eight male or female subjects were randomized within each dose panel (1, 10, 25, 50, 100 and 200mg) to drug or placebo in a ratio of 3:1. BMS-790052 or placebo was administered in the fasted state. The plasma samples obtained at various times were analysed for BMS-790052 by a validated liquid chromatography tandem mass spectrometry assay. Pharmacokinetic parameter values for individual subjects were derived by non-compartmental methods by a validated pharmacokinetic analysis programme. PBA EC90, protein-binding-adjusted EC90 for the individual genotype in a replicon assay. Error bars, standard deviation.

  4. Mean change in log10 HCV RNA with 90% confidence intervals after administration of single oral doses of BMS-790052 to HCV-infected patients.
    Figure 4: Mean change in log10 HCV RNA with 90% confidence intervals after administration of single oral doses of BMS-790052 to HCV-infected patients.

    In a double-blind, placebo-controlled, sequential, single ascending-dose study, six subjects were randomized within each dose panel (1, 10, 100mg) to drug or placebo in a ratio of 5:1. BMS-790052 or placebo was administered in the fasted state. Owing to a dosing error, all six subjects received BMS-790052 in the 1mg panel. One subject in the 10mg panel withdrew from the study 8h after administration of the study drug for non-drug-related reasons; HCV RNA data from the subject are included up until the subject withdrew.

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Author information

Affiliations

  1. Department of Virology, Bristol-Myers Squibb Research and Development, 5 Research Parkway, Wallingford, Connecticut 06492, USA

    • Min Gao,
    • Robert A. Fridell,
    • Jin-Hua Sun,
    • Donald R. O’Boyle II,
    • Julie A. Lemm,
    • Chunfu Wang &
    • Richard J. Colonno
  2. Department of Discovery Medicine and Clinical Pharmacology, Bristol-Myers Squibb Research and Development, Princeton, New Jersey 08543, USA

    • Richard E. Nettles,
    • Caly Chien &
    • Dennis M. Grasela
  3. Department of Discovery Chemistry,

    • Makonen Belema,
    • Lawrence B. Snyder,
    • Van N. Nguyen,
    • Michael H. Serrano-Wu,
    • Nicholas A. Meanwell &
    • Lawrence G. Hamann
  4. Department of Computer-Aided Drug Design,

    • David R. Langley
  5. Department of Metabolism and Pharmacokinetics, Bristol-Myers Squibb Research and Development, 5 Research Parkway, Wallingford, Connecticut 06492, USA

    • Jay O. Knipe

Contributions

R.A.F., J.A.L., D.R.O., J.-H.S., C.W. and M.G. designed and performed the replicon screen, constructed replicons and hybrid replicons, conducted genotype coverage and inhibitor-binding experiments, isolated and mapped resistant variants, designed in vitro combination studies and performed genotypic and phenotypic analysis of clinical samples. M.G., R.A.F., J.A.L. and R.J.C. designed the overall virology studies and provided input to the overall research direction. J.O.K. designed and interpreted in vivo pharmacokinetic studies and in vitro compound profiling. M.B., V.N.N., L.B.S. and M.H.S.-W. designed and synthesized the compounds, L.B.S., L.G.H. and N.A.M. provided direction to the chemistry research and contributed to the design of compounds. D.R.L. constructed models of HCV NS5A and contributed to the development of mechanistic hypotheses. R.E.N., C.C. and D.M.G. designed and interpreted human trials, and D.M.G. provided input to the overall research direction. M.G., R.E.N. and N.A.M. co-authored the manuscript with input from all co-authors, and N.A.M. compiled and edited the article.

Competing financial interests

The authors are or were, at the time this work was conducted, employees of Bristol-Myers Squibb.

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  1. Supplementary Information (401K)

    This file contains Supplementary Tables S1-S14, Supplementary References and data for Experimental Chemistry.

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