Proteomic characterisation of drug metabolising enzymes and drug transporters in pig liver

Abstract

1. Liver enzymes and transporters play an essential role in xenobiotic metabolism, distribution and elimination. Pre-clinical safety assessment relies on studies on animal models, including the (mini)pig. The pig shares many anatomical and physiological characteristics with humans, and there is currently a gap in information about porcine metabolism and disposition pathways and their similarities and differences from human ones.

2. Three different sample preparation methods (filter-aided sample preparation (FASP), enhanced FASP (eFASP) and in-solution sample preparation) were used to prepare porcine liver tissue (two samples) for proteomic analysis. The analysis relied on rapid-separation liquid chromatography coupled to Orbitrap mass spectrometry in data-dependent acquisition mode. MASCOT was used for identification and relative label-free quantification was based on spectral counting.

3. The three sample preparation methods provided complementary results, allowing characterisation of approximately 70 pharmacologically relevant proteins. The main quantified proteins included 16 cytochrome P450 (CYP) enzymes, 5 UGT enzymes, and 11 transporters. In addition, 20 Phase I and 14 Phase II enzymes were also characterised. Inter-operator differences were negligible and the pig liver pies for CYP, UGT and efflux transporter proteins were established. Human homologues of the quantified CYP, UGT and transporter proteins were identified.

Keywords:

• Enzymes
• transporters
• proteomics
• pig
• quantification

Acknowledgements

The authors thank the Biological Mass Spectrometry BioMS Core Facility, University of Manchester, for access to mass spectrometry instruments, the Egyptian Government for sponsoring Yasmine Elmorsi and CAPKR consortium (Janssen, Certara, Merck, MSD, Genentech, Takeda Pharmaceuticals and Eli Lilly) for support.

Disclosure statement

The authors report no declarations of interest.