1. 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
- Author
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David J. Elliot, Susan J. Semple, Matthew Y Bendikov, John O. Miners, Matthew J. Sykes, Bradley S Simpson, Elizabeth M. J. Gillam, Ross A. McKinnon, David J. Claudie, Bendikov, Matthew Y, Miners, John O, Simpson, Bradley S, Elliot, David J, Semple, Susan J, Claudie, David J, McKinnon, Ross A, Gillam, Elizabeth MJ, and Sykes, Matthew J
- Subjects
0301 basic medicine ,Glucuronosyltransferase ,cytochrome P450 ,Health, Toxicology and Mutagenesis ,Metabolite ,Glucuronidation ,Anti-Inflammatory Agents ,Toxicology ,digestive system ,030226 pharmacology & pharmacy ,Biochemistry ,Diterpenes, Clerodane ,esterases ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Glucuronides ,Cytochrome P-450 Enzyme System ,polyandric acid A ,Humans ,reaction phenotyping ,Pharmacology ,chemistry.chemical_classification ,clerodane diterpenoid ,biology ,glucuronidation ,Australia ,Cytochrome P450 ,General Medicine ,UGT2B7 ,030104 developmental biology ,Enzyme ,chemistry ,Microsome ,biology.protein ,Microsomes, Liver ,in vitro–in vivo extrapolation ,Glucuronide ,Oxidation-Reduction ,UDP-glucuronosyltransferase - 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. Refereed/Peer-reviewed
- Published
- 2016