Biotechnological Synthesis of the Designer Drug Metabolite 4'-Hydroxymethyl-a-pyrrolidinohexanophenone in Fission Yeast Heterologously Expressing Human Cytochrome P450 2D6―A Versatile Alternative to Multistep Chemical Synthesis

1-(4-Methylphenyl)-2-pyrrolidin-1-ylhexan-1-one (4'-methyl-a-pyrrolidinohexanophenone, MPHP) is a new designer drug that appeared on the illicit drug market. It is mainly metabolized to 4'-hydroxymethyl-a-pyrrolidinohexanophenone (HO-MPHP) followed by oxidation to the respective carboxylic...

Full description

Saved in:
Bibliographic Details
Published in:Journal of analytical toxicology 2009-05, Vol.33 (4), p.190-197
Main Authors: PETERS, Frank T, DRAGAN, Calin-Aurel, KAUFFELS, Anne, SCHWANINGER, Andrea E, ZAPP, Josef, BUREIK, Matthias, MAURER, Hans H
Format: Article
Language:English
Subjects:
Biological and medical sciences
Drug addictions
Medical sciences
Schizosaccharomyces pombe
Toxicology
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:1-(4-Methylphenyl)-2-pyrrolidin-1-ylhexan-1-one (4'-methyl-a-pyrrolidinohexanophenone, MPHP) is a new designer drug that appeared on the illicit drug market. It is mainly metabolized to 4'-hydroxymethyl-a-pyrrolidinohexanophenone (HO-MPHP) followed by oxidation to the respective carboxylic acid. For studies on the quantitative involvement of human cytochrome P450 (CYP) isoenzymes in the initial hydroxylation, a reference standard of HO-MPHP was needed. Therefore, the aim of this study was to synthesize this metabolite using a biotechnological approach. MPHP·HNO3 (250 µmol) was incubated with 1 L culture of the fission yeast (Schizosaccharomyces pombe) strain CAD64 heterologously co-expressing human CYP reductase and CYP2D6. After centrifugation, the product was isolated from the incubation supernatants by solid-phase extraction. Further product cleanup was achieved by semi-preparative high-performance liquid chromatography (HPLC). After extraction of HO-MPHP from the respective eluent fractions, it was precipitated as its hydrochloric salt. The final product HO-MPHP·HCl was obtained in a yield of 138 µmol (43 mg, 55%). Its identity was confirmed by full scan gas chromatography-mass spectrometry (after trimethylsilylation), 1H-NMR, and 13C-NMR. The product purity as estimated from HPLC-ultraviolet analysis was greater than 99%. The described biotechnological approach proved to be a versatile alternative to the chemical synthesis of HO-MPHP.
ISSN:0146-4760
1945-2403
DOI:10.1093/jat/33.4.190