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Parameterization of Microsomal and Cytosolic Scaling Factors: Methodological and Biological Considerations for Scalar Derivation and Validation

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Abstract

Mathematical models that can predict the kinetics of compounds have been increasingly adopted for drug development and risk assessment. Data for these models may be generated from in vitro experimental systems containing enzymes contributing to metabolic clearance, such as subcellular tissue fractions including microsomes and cytosol. Extrapolation from these systems is facilitated by common scaling factors, known as microsomal protein per gram (MPPG) and cytosolic protein per gram (CPPG). Historically, parameterization of MPPG and CPPG has employed the use of recovery factors, commonly benchmarked to cytochromes P450 which work well in some contexts, but could be problematic for other enzymes. Here, we propose absolute quantification of protein content and supplementary assays to evaluate microsomal/cytosolic purity that should be employed. Examples include calculation of microsomal latency by mannose-6-phosphatase activity and immunoblotting of subcellular fractions with fraction-specific markers. Further considerations include tissue source, as disease states can affect enzyme expression and activity, and the methodology used for scalar parameterization. Regional- and organ-specific expression of enzymes, in addition to differences in organ physiology, is another important consideration. Because most efforts have focused on the liver that is, for the most part, homogeneous, derived scalars may not capture the heterogeneity of other major tissues contributing to xenobiotic metabolism including the kidneys and small intestine. Better understanding of these scalars, and how to appropriately derive them from extrahepatic tissues can provide support to the inferences made with physiologically based pharmacokinetic modeling, increase its accuracy in characterizing in vivo drug pharmacokinetics, and improve confidence in go-no-go decisions for clinical trials.

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  1. Faculty of Pharmaceutical Sciences, The University of British Columbia, 2405 Wesbrook Mall, Vancouver, BC, V6T 1Z3, Canada

    Michael J. Doerksen, Michael W. H. Coughtrie & Abby C. Collier

  2. Genentech Inc, 1 DNA Way, South San Francisco, California, 94080, USA

    Robert S. Jones

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  1. Michael J. Doerksen
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Correspondence to Abby C. Collier.

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MJD is supported by an Alexander Graham Bell Canada Graduate Scholarship from the National Science and Engineering Council of Canada (NSERC), and the Frank S. Abbott Graduate Fellowship in Pharmaceutical Sciences. ACC holds research grants from the NSERC Discovery Grants Program (17-3808), and a Genentech Research Grant that supported this work.

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Doerksen, M.J., Jones, R.S., Coughtrie, M.W.H. et al. Parameterization of Microsomal and Cytosolic Scaling Factors: Methodological and Biological Considerations for Scalar Derivation and Validation. Eur J Drug Metab Pharmacokinet 46, 173–183 (2021). https://doi.org/10.1007/s13318-020-00666-w

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