Uncoupling of Cytochrome P450 2B6 and stimulation of reactive oxygen species production in pharmacogenomic alleles affected by interethnic variability

• Published in: Biochimica et biophysica acta. General subjects Vol. 1868; no. 5; p. 130595
• Main Authors: Yamoune, Sabrina, Müller, Julian Peter, Langmia, Immaculate Mbongo, Scholl, Catharina, Stingl, Julia Carolin
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2024
• Subjects: Alleles; amino acids; biochemical pathways; biotransformation; Bupropion - metabolism; Bupropion - pharmacology; CYP2B6; CYP2B66; cytochrome P-450; Cytochrome P-450 CYP2B6 - drug effects; Cytochrome P-450 CYP2B6 - genetics; dissociation; enzyme activity; functional diversity; Humans; Hydrogen Peroxide; ketamine; Ketamine - metabolism; Ketamine - pharmacology; peroxidase; Peroxide shunt; Pharmacogenetics; Pharmacogenomics; pharmacokinetics; Reactive Oxygen Species; ROS; Tandem Mass Spectrometry; Uncoupling; Uncoupling; KETA; Peroxide shunt; CYP2B6; EFV; CYP2B66; CYP450; ROS; HB; BUP; HE; NK; Pharmacogenomics
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2024.130595

Cytochrome P450 mediated substrate metabolism is generally characterized by the formation of reactive intermediates. In vitro and in vivo reaction uncoupling, results in the accumulation and dissociation of reactive intermediates, leading to increased ROS formation. The susceptibility towards uncoupling and altered metabolic activity is partly modulated by pharmacogenomic alleles resulting in amino acid substitutions. A large variability in the prevalence of these alleles has been demonstrated in CYP2B6, with some being predominantly unique to African populations. The aim of this study is to characterize the uncoupling potential of recombinant CYP2B6*1, CYP2B6*6 and CYP2B6*34 metabolism of specific substrates. Therefore, functional effects of these alterations on enzyme activity were determined by quantification of bupropion, efavirenz and ketamine biotransformation using HPLC-MS/MS. Determination of H2O2 levels was performed by the AmplexRed/horseradish peroxidase assay. Our studies of the amino acid substitutions Q172H, K262R and R487S revealed an exclusive use of the peroxide shunt for the metabolism of bupropion and ketamine by CYP2B6*K262R. Ketamine was also identified as a trigger for the peroxide shunt in CYP2B6*1 and all variants. Concurrently, ketamine acted as an uncoupler for all enzymes. We further showed that the expressed CYP2B6*34 allele results in the highest H2O2 formation. We therefore conclude that the reaction uncoupling and peroxide shunt are directly linked and can be substrate specifically induced with K262R carriers being most likely to use the peroxide shunt and R487S carrier being most prone to reaction uncoupling. This elucidates the functional diversity of pharmacogenomics in drug metabolism and safety. [Display omitted] •CYP2B6*6 enzyme shows greater bupropion metabolism in vitro compared to CYP2B6*1.•CYP2B6*6 and CYP2B6*34 enzymes show higher H2O2 production in vitro.•H2O2 production is reduced in presence of bupropion and ketamine, not efavirenz.•K262R and R487S subsitutions are more prone to use the H2O2-shunt.•Ketamine induces the metabolism via the H2O2-shunt in CYP2B6*1 and CYP2B6 variants.