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Applicability of in vitro-in vivo translation of cathepsin K inhibition from animal species to human with the use of free-drug hypothesis

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Abstract

The correlation of in vitro inhibition of cathepsin K (CatK) activity and in vivo suppression of collagen I biomarkers was examined with three selective CatK inhibitors to explore the potential translatability from animal species to human. These inhibitors exhibited good in vitro potencies toward recombinant CatK enzymes across species, with IC50 values ranging from 0.20 to 6.1 nM. In vivo studies were conducted in animal species following multiple-day dosing of the CatK inhibitors to achieve steady-state plasma drug concentration-time profiles. Measurement of urinary bone resorption biomarkers (cross-linked N-terminal telopeptide and helical peptide of type I collagen) revealed drug concentration-dependent suppression of biomarkers, with EC50 values estimated to be 12 to 160 nM. Marked improvement in the correlation between in vitro and in vivo CatK activities was observed with the application of unbound (free) fraction in plasma, consistent with the conditions stipulated by the free-drug hypothesis. These results indicate that the in vitro-in vivo translation of CatK inhibition observed in animal species can translate to humans when the unbound fraction of the inhibitor is considered. Interestingly, residual levels of urinary bone resorption marker were detected as the suppression reached saturation (at an average of 82% inhibition), an apparent phenomenon observed regardless of the species, biomarker, or compound examined. Since cathepsin enzymes other than CatK were reported to catalyze cleavage of collagen I, it is hypothesized that CatK-mediated degradation of collagen I in bone represents ~82% of overall collagen I turnover in the body.

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Acknowledgements

The authors thank Peter Szczerba and Rachel Korn for conducting measurements of urinary biomarkers, Li Sun and Joan Ellis for providing quantification of CatK inhibitors in plasma, Dr. Michael Gentile for providing additional data from the ovariectomized-rabbit study with odanacatib, and Dr. Stefan Zajic and Dr. Julie Stone for additional human PK/PD analyses and valuable discussion.

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Authors and Affiliations

  1. Department of Pharmacokinetics, Pharmacodynamics and Drug Metabolism, MSD, West Point, PA, 19486, USA

    Bennett Ma

  2. Department of Pharmacology, MSD, West Point, PA, USA

    Bin Luo, Danielle H. Euler, Yangsi Ou, Steven S. Carroll & Laura S. Lubbers

  3. Department of Bone Biology, MSD, West Point, PA, USA

    Tara E. Cusick, Gregg Wesolowski, Helmut Glantschnig & Le T. Duong

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  1. Bennett Ma
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  2. Bin Luo
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  3. Danielle H. Euler
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Correspondence to Bennett Ma.

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All authors are current or former employees of MSD who were employed at the time the experiment was conducted. The authors alone are responsible for the writing and content of this article.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Ma, B., Luo, B., Euler, D.H. et al. Applicability of in vitro-in vivo translation of cathepsin K inhibition from animal species to human with the use of free-drug hypothesis. Naunyn-Schmiedeberg's Arch Pharmacol 390, 435–441 (2017). https://doi.org/10.1007/s00210-017-1356-5

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