Metabolism of 1,8-cineole by human cytochrome P450 enzymes: identification of a new hydroxylated metabolite

• Published in: Biochimica et biophysica acta. General subjects 2005-04, Vol.1722 (3), p.304-311
• Main Authors: Duisken, Mike, Sandner, Frank, Blömeke, Brunhilde, Hollender, Juliane
• Format: Article
• Language: English
• Subjects: 1,8-Cineole; 2α-Hydroxy-1,8-cineole; 3α-Hydroxy-1,8-cineole; CYP3A; Cytochrome P450; Metabolism
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2004.12.019

Human metabolism of the monoterpene cyclic ether 1,8-cineole was investigated in vitro and in vivo. In vitro, the biotransformation of 1,8-cineole was investigated by human liver microsomes and by recombinant cytochrome P450 enzymes coexpressed with human CYP-reductase in Escherichia coli cells. Besides the already described metabolite 2α-hydroxy-1,8-cineole we found another metabolite produced at high rates. The structure was identified by a comparison of its mass spectrum and retention time with the reference compounds as 3α-hydroxy-1,8-cineole. There was a clear correlation between the concentration of the metabolites, incubation time and enzyme content, respectively. CYP3A4/5 antibody significantly inhibited the 2α- and 3α-hydroxylation catalyzed by pooled human liver microsomes. Further kinetic analysis revealed that the Michaelis–Menten K m and V max for oxidation of 1,8-cineole in position three were 19 μM and 64.5 nmol/min/nmol P450 for cytochrome P450 3A4, and 141 μM and 10.9 nmol/min/nmol P450 for cytochrome P450 3A5, respectively. To our knowledge, this is the first time that 3α-hydroxy-1,8-cineole is described as a human metabolite of 1,8-cineole. We confirmed these in vitro results by the investigation of human urine after the oral administration of cold medication containing 1,8-cineole. In human urine we found by GC-MS analysis the described metabolites, 2α-hydroxy-1,8-cineole and 3α-hydroxy-1,8-cineole.