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Drug-induced cholestasis assay in primary hepatocytes

Drug-induced cholestasis (DIC) is a major cause of clinical failure of drug candidates. Numerous patients worldwide are affected when exposed to marketed drugs exhibiting a DIC signature. Prospective identification of DIC during early compound development remains challenging. Here we describe the op...

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Autores principales: Van Brantegem, Pieter, Chatterjee, Sagnik, De Bruyn, Tom, Annaert, Pieter, Deferm, Neel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559231/
https://www.ncbi.nlm.nih.gov/pubmed/33088729
http://dx.doi.org/10.1016/j.mex.2020.101080
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author Van Brantegem, Pieter
Chatterjee, Sagnik
De Bruyn, Tom
Annaert, Pieter
Deferm, Neel
author_facet Van Brantegem, Pieter
Chatterjee, Sagnik
De Bruyn, Tom
Annaert, Pieter
Deferm, Neel
author_sort Van Brantegem, Pieter
collection PubMed
description Drug-induced cholestasis (DIC) is a major cause of clinical failure of drug candidates. Numerous patients worldwide are affected when exposed to marketed drugs exhibiting a DIC signature. Prospective identification of DIC during early compound development remains challenging. Here we describe the optimized in vitro • Exposure of primary human hepatocyte cultures to test compounds in the absence and presence of a physiologically relevant mixture of endogenous bile salts. • Rapid and quantitative assessment of the influence of concomitant bile salt exposure on hepatocyte functionality and integrity after 24 h or 48 h of incubation. • Translation of the in vitro result, expressed as a DIC index (DICI) value, into an in vivo safety margin. Using our historical control data, a new (data driven) DICI cut-off value of 0.78 was established for discerning cholestatic and non-cholestatic compounds. Our DIC assay protocol was further improved by now relying on the principle of the no observable adverse effect level (NOAEL) for determining the highest test compound concentration corresponding to a DICI  ≥  0.78. Predicted safety margin values were subsequently calculated for compounds displaying hepatotoxic and/or cholestatic effects in patients, thus enabling evaluation of the performance of our DIC assay. Of note, this assay can be extended to explore the role of drug metabolites in precipitating DIC.
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spelling pubmed-75592312020-10-20 Drug-induced cholestasis assay in primary hepatocytes Van Brantegem, Pieter Chatterjee, Sagnik De Bruyn, Tom Annaert, Pieter Deferm, Neel MethodsX Method Article Drug-induced cholestasis (DIC) is a major cause of clinical failure of drug candidates. Numerous patients worldwide are affected when exposed to marketed drugs exhibiting a DIC signature. Prospective identification of DIC during early compound development remains challenging. Here we describe the optimized in vitro • Exposure of primary human hepatocyte cultures to test compounds in the absence and presence of a physiologically relevant mixture of endogenous bile salts. • Rapid and quantitative assessment of the influence of concomitant bile salt exposure on hepatocyte functionality and integrity after 24 h or 48 h of incubation. • Translation of the in vitro result, expressed as a DIC index (DICI) value, into an in vivo safety margin. Using our historical control data, a new (data driven) DICI cut-off value of 0.78 was established for discerning cholestatic and non-cholestatic compounds. Our DIC assay protocol was further improved by now relying on the principle of the no observable adverse effect level (NOAEL) for determining the highest test compound concentration corresponding to a DICI  ≥  0.78. Predicted safety margin values were subsequently calculated for compounds displaying hepatotoxic and/or cholestatic effects in patients, thus enabling evaluation of the performance of our DIC assay. Of note, this assay can be extended to explore the role of drug metabolites in precipitating DIC. Elsevier 2020-09-25 /pmc/articles/PMC7559231/ /pubmed/33088729 http://dx.doi.org/10.1016/j.mex.2020.101080 Text en © 2020 The Author(s). Published by Elsevier B.V. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Method Article
Van Brantegem, Pieter
Chatterjee, Sagnik
De Bruyn, Tom
Annaert, Pieter
Deferm, Neel
Drug-induced cholestasis assay in primary hepatocytes
title Drug-induced cholestasis assay in primary hepatocytes
title_full Drug-induced cholestasis assay in primary hepatocytes
title_fullStr Drug-induced cholestasis assay in primary hepatocytes
title_full_unstemmed Drug-induced cholestasis assay in primary hepatocytes
title_short Drug-induced cholestasis assay in primary hepatocytes
title_sort drug-induced cholestasis assay in primary hepatocytes
topic Method Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559231/
https://www.ncbi.nlm.nih.gov/pubmed/33088729
http://dx.doi.org/10.1016/j.mex.2020.101080
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