Biotransformation, Using Recombinant CYP450-Expressing Baker's Yeast Cells, Identifies a Novel CYP2D6.10 Variant Which Is a Superior Metabolizer of Codeine to Morphine Than the Wild-Type Enzyme.

Clicks: 317
ID: 52723
2018
CYP2D6, a cytochrome P450 (CYP) enzyme, metabolizes codeine to morphine. Within the human body, 0-15% of codeine undergoes O-demethylation by CYP2D6 to form morphine, a far stronger analgesic than codeine. Genetic polymorphisms in wild-type CYP2D6 (CYP2D6-wt) are known to cause poor-to-extensive metabolism of codeine and other CYP2D6 substrates. We have established a platform technology that allows stable expression of human CYP genes from chromosomal loci of baker's yeast cells. Four CYP2D6 alleles, (i) chemically synthesized CYP2D6.1, (ii) chemically synthesized CYP2D6-wt, (iii) chemically synthesized CYP2D6.10, and (iv) a novel CYP2D6.10 variant CYP2D6-C (i.e., CYP2D6.10) isolated from a liver cDNA library, were cloned for chromosomal integration in yeast cells. When expressed in yeast, CYP2D6.10 enzyme shows weak activity compared with CYP2D6-wt and CYP2D6.1 which have moderate activity, as reported earlier. Surprisingly, however, the CYP2D6-C enzyme is far more active than CYP2D6.10. More surprisingly, although CYP2D6.10 is a known low metabolizer of codeine, yeast cells expressing CYP2D6-C transform >70% of codeine to morphine, which is more than twice that of cells expressing the extensive metabolizers, CYP2D6.1, and CYP2D6-wt. The latter two enzymes predominantly catalyze formation of codeine's N-demethylation product, norcodeine, with >55% yield. Molecular modeling studies explain the specificity of CYP2D6-C for O-demethylation, validating observed experimental results. The yeast-based CYP2D6 expression systems, described here, could find generic use in CYP2D6-mediated drug metabolism and also in high-yield chemical reactions that allow the formation of regio-specific dealkylation products.
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williams2018biotransformationacs Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Williams, Ibidapo S;Gatchie, Linda;Bharate, Sandip B;Chaudhuri, Bhabatosh;
Journal ACS omega
Year 2018
DOI 10.1021/acsomega.8b00809
URL
Keywords
pharmacokinetics poloxamer sustained release in vitro release sd, standard deviation im, intramuscular atp, adenosine 5′ triphosphate auc80h, area under the analyte concentration versus time curve from time zero to 80 h auclast, area under the analyte concentration versus time curve from time zero to the time of the last measurable (non-below quantification level) concentration auc∞, area under the analyte concentration vs time curve from time zero to infinite time c0, analyte plasma concentration at time zero can, acetonitrile cmc, critical micellar concentration cmt, critical micellar temperature cmax, maximum observed analyte plasma concentration dn, dose normalized doe, design of experiments eo, ethylene oxide gpt, gel point temperature gel erosion h&e, hematoxylin and eosin iv, intravenous in situ forming gels k2.edta, potassium ethylenediaminetetraacetic acid lc–ms/ms, liquid chromatography-tandem mass spectrometry mdr-tb, multi-drug resistant tuberculosis mrm, multiple reaction monitoring nmp, n-methyl-2-pyrrolidone p338, poloxamer 338 p407, poloxamer 407 plga, poly-(dl-lactic-co-glycolic acid) po, propylene oxide tfa, trifluoroacetic acid uhplc, ultra-high performance liquid chromatography uv, ultraviolet n, sample size t1/2, apparent terminal elimination half-life tlast, sampling time until the last measurable (non-below quantification level) analyte plasma concentration tmax, sampling time to reach the maximum observed analyte plasma concentration

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