title: In vitro metabolism of the anti-inflammatory clerodane diterpenoid polyandric acid A and its hydrolysis product by human liver microsomes and recombinant cytochrome P450 and UDP-glucuronosyltransferase enzymes.
abstract: 1. The metabolism of the anti-inflammatory diterpenoid polyandric acid A (PAA), a constituent of the Australian Aboriginal medicinal plant Dodonaea polyandra, and its de-esterified alcohol metabolite, hydrolysed polyandric acid A (PAAH) was studied in vitro using human liver microsomes (HLM) and recombinant UDP-glucuronosyltransferase (UGT) and cytochrome P450 (CYP) enzymes. 2. Hydrolysis of PAA to yield PAAH occurred upon incubation with HLM. Further incubations of PAAH with HLM in the presence of UGT and CYP cofactors resulted in significant depletion, with UGT-mediated depletion as the major pathway. 3. Reaction phenotyping utilising selective enzyme inhibitors and recombinant human UGT and CYP enzymes revealed UGT2B7 and UGT1A1, and CYP2C9 and CYP3A4 as the major enzymes involved in the metabolism of PAAH. 4. Analysis of incubations of PAAH with UDP-glucuronic acid-supplemented HLM and recombinant enzymes by UPLC/MS/MS identified three glucuronide metabolites. The metabolites were further characterised by β-glucuronidase and mild alkaline hydrolysis. The acyl glucuronide of PAAH was shown to be the major metabolite. 5. This study demonstrates the in vitro metabolism of PAA and PAAH and represents the first systematic study of the metabolism of an active constituent of an Australian Aboriginal medicinal plant.
authors.affiliation: a Department of Clinical Pharmacology , School of Medicine, Flinders University , Adelaide , Australia.|b Centre for Drug Discovery and Development, Sansom Institute for Health Research, University of South Australia , Adelaide , Australia.
authors.forename: Matthew Y
authors.initials: MY
authors.lastname: Bendikov
authors.affiliation: a Department of Clinical Pharmacology , School of Medicine, Flinders University , Adelaide , Australia.|c Flinders Centre for Innovation in Cancer, School of Medicine, Flinders University , Adelaide , Australia.
authors.forename: John O
authors.initials: JO
authors.lastname: Miners
authors.affiliation: c Flinders Centre for Innovation in Cancer, School of Medicine, Flinders University , Adelaide , Australia.|d Quality Use of Medicines and Pharmacy Research Centre, Sansom Institute for Health Research, University of South Australia , Adelaide , Australia.
authors.forename: Bradley S
authors.initials: BS
authors.lastname: Simpson
authors.affiliation: e Department of Clinical Pharmacology , Flinders Medical Centre , Adelaide , Australia.
authors.forename: David J
authors.initials: DJ
authors.lastname: Elliot
authors.affiliation: d Quality Use of Medicines and Pharmacy Research Centre, Sansom Institute for Health Research, University of South Australia , Adelaide , Australia.
authors.forename: Susan J
authors.initials: SJ
authors.lastname: Semple
authors.affiliation: f Chuulangun Aboriginal Corporation , Cairns Mail Centre , Cairns , Australia , and.
authors.forename: David J
authors.initials: DJ
authors.lastname: Claudie
authors.affiliation: c Flinders Centre for Innovation in Cancer, School of Medicine, Flinders University , Adelaide , Australia.
authors.forename: Ross A
authors.initials: RA
authors.lastname: McKinnon
authors.affiliation: g School of Chemistry and Molecular Biosciences, the University of Queensland , Brisbane , Australia.
authors.forename: Elizabeth M J
authors.initials: EMJ
authors.lastname: Gillam
authors.affiliation: b Centre for Drug Discovery and Development, Sansom Institute for Health Research, University of South Australia , Adelaide , Australia.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Sykes
mesh_terms: D000893:Anti-Inflammatory Agents / Q000378:metabolism*; D001315:Australia; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D045785:Diterpenes, Clerodane / Q000378:metabolism*; D020719:Glucuronides / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000378:metabolism; D006801:Humans; D008862:Microsomes, Liver / Q000378:metabolism*; D010084:Oxidation-Reduction; D000099148:UGT1A1 Enzyme
chemical_list: D000893:Anti-Inflammatory Agents; D003577:Cytochrome P-450 Enzyme System; D045785:Diterpenes, Clerodane; D020719:Glucuronides; D014453:Glucuronosyltransferase; D000099148:UGT1A1 Enzyme; C587407:polyandric acid A
keywords: Clerodane diterpenoid; UDP-glucuronosyltransferase; cytochrome P450; esterases; glucuronidation; in vitro–in vivo extrapolation; polyandric acid A; reaction phenotyping