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In vitro Phase I- and Phase II-Drug Metabolism in The Liver of Juvenile and Adult Göttingen Minipigs

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Abstract

Purpose

In view of pediatric drug development, juvenile animal studies are gaining importance. However, data on drug metabolizing capacities of juvenile animals are scarce, especially in non-rodent species. Therefore, we aimed to characterize the in vitro biotransformation of four human CYP450 substrates and one UGT substrate in the livers of developing Göttingen minipigs.

Methods

Liver microsomes from late fetal, Day 1, Day 3, Day 7, Day 28, and adult male and female Göttingen minipigs were incubated with a cocktail of CYP450 substrates, including phenacetin, tolbutamide, dextromethorphan, and midazolam. The latter are probe substrates for human CYP1A2, CYP2C9, CYP2D6, and CYP3A4, respectively. In addition, the UGT multienzyme substrate (from the UGT-GloTM assay), which is glucuronidated by several human UGT1A and UGT2B enzymes, was also incubated with the porcine liver microsomes.

Results

For all tested substrates, drug metabolism significantly rose postnatally. At one month of age, 60.5 and 75.4% of adult activities were observed for acetaminophen and dextrorphan formations, respectively, while 35.4 and 43.2% of adult activities were present for 4-OH-tolbutamide and 1’-OH-midazolam formations. Biotransformation of phenacetin was significantly higher in 28-day-old and adult females compared with males.

Conclusions

Maturation of metabolizing capacities occurred postnatally, as described in man.

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Abbreviations

CYP450:

Cytochrome P450 enzymes

DGA:

Days of gestational age

HLM:

Human liver microsomes

KPO4 :

Potassium phosphate

LLOQ:

Lower limit of quantification

MLM:

Minipig liver microsomes

mol/min/mg MP:

Moles of metabolite formed per minute per milligram of microsomal protein

MP:

Microsomal protein

PBS:

Phosphate buffered saline

PND:

Postnatal day

RLU:

Relative light unit

RT:

Room temperature

TBS:

Tris buffered saline

UDPGA:

Uridine-5’-diphospho-glucuronic acid

UGT:

Uridine diphosphate glucuronosyltransferase

Vmax :

Maximal velocity

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ACKNOWLEDGMENTS AND DISCLOSURES

The Applied Veterinary Morphology research group would like to thank Ellegaard Göttingen Minipig A/S for the kind donation of animals. The authors from the University of Antwerp are members of COST Action BM1308 ‘Sharing Advances on Large Animal Models (SALAAM)’.

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Authors and Affiliations

  1. Laboratory of Applied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610, Wilrijk, Belgium

    Els Van Peer, Christophe Casteleyn, Chris Van Ginneken & Steven Van Cruchten

  2. Janssen R&D, Beerse, Belgium

    Frank Jacobs, Jan Snoeys, Jos Van Houdt & Ils Pijpers

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  1. Els Van Peer
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Correspondence to Steven Van Cruchten.

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The authors declare no conflict of interest.

Ethical Approval

The Ethical Committee of Animal Experimentation from the University of Antwerp (Belgium) approved the protocol and use of the animals and research adhered to the ‘Principles of Laboratory Animal Care’ (NIH publication Nr 85-23, revised in 1985).

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Van Peer, E., Jacobs, F., Snoeys, J. et al. In vitro Phase I- and Phase II-Drug Metabolism in The Liver of Juvenile and Adult Göttingen Minipigs. Pharm Res 34, 750–764 (2017). https://doi.org/10.1007/s11095-017-2101-y

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  • DOI: https://doi.org/10.1007/s11095-017-2101-y

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