Metabolism of ∆-Carene by Human Cytochrome P450 Enzymes: Identification and Characterization of Two New Metabolites

نویسندگان

  • Mike Duisken
  • Dorina Benz
  • Thomas H. Peiffer
  • Brunhilde Blömeke
  • Juliane Hollender
چکیده

The metabolism of the bicyclic monoterpene ∆-carene was investigated in vitro using human liver microsomes as well as human smoker/non-smoker lung microsomes and 12 different recombinant cytochrome P450 enzymes coexpressed with human CYP-reductase in Escherichia coli cells. We detected two metabolites using GC-MS analysis. The mass fragmentation indicated for one metabolite hydroxylation in the allyl position and for the other metabolite epoxidation at the double bond. For clear identification the suggested metabolites were synthesized in a four-step reaction. Comparison of GC retention times and mass spectra lead to the identification of the metabolites as ∆-carene-10-ol ((1S, 6R)7,7-Dimethylbicyclo[4.1.0]hept-3-en-3-yl-methanol) and ∆-carene-epoxide ((1S, 3S, 5R, 7R)-3,8,8-Trimethyl-4-oxatricyclo[5.1.0.0]octane). ∆-carene-10-ol was formed by human liver microsomes and recombinant human CYP2B6, CYP2C19 and CYP2D6. ∆-Carene-epoxide was obviously catalyzed only by CYP1A2. In both cases there was a clear correlation between the metabolite formation, incubation time and enzyme concentration, respectively. Further kinetic analysis revealed that CYP2B6 exhibited the highest activity for ∆-carene 10-hydroxylation. Michaelis-Menten Km and Vmax for oxidation of ∆-carene were 0.6 mM and 28.4 nmol/min/nmol P450 using human CYP2B6. For the formation of ∆-carene-epoxide 98.2 mM and 3.9 nmol/min/nmol P450 were determined as Km and Vmax by using human CYP1A2. To our knowledge, this is the first time that ∆-carene-10-ol and ∆-carene-epoxide are described as human metabolites of ∆-

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Expression of cytochrome P450 and glutathione S-transferase in human bone marrow mesenchymal stem cells

Currently several studies are being carried out on various properties of mesenchymal stem cells (MSCs)however there are a few investigations about drug metabolizing properties of these cells. The aim of thisstudy was to measure the key factors involved in drug metabolism in human bone marrow MSCs. For thispurpose, cellular glutathione (GSH), glutathione Stransferase (GSTs) and...

متن کامل

In vitro identification of the P450 enzymes responsible for the metabolism of ropinirole.

The in vitro metabolism of ropinirole was investigated with the aim of identifying the cytochrome P450 enzymes responsible for its biotransformation. The pathways of metabolism after incubation of ropinirole with human liver microsomes were N-despropylation and hydroxylation. Enzyme kinetics demonstrated the involvement of at least two enzymes contributing to each pathway. A high affinity compo...

متن کامل

Identification of enzymes responsible for primary and sequential oxygenation reactions of capravirine in human liver microsomes.

Capravirine, a new non-nucleoside reverse transcriptase inhibitor, undergoes extensive oxygenation reactions, including N-oxidation, sulfoxidation, sulfonation, and hydroxylation in humans. Numerous primary (mono-oxygenated) and sequential (di-, tri-, and tetraoxygenated) metabolites of capravirine are formed via the individual or combined oxygenation pathways. In this study, cytochrome P450 en...

متن کامل

Prevalence of Non-Cytochrome P450-Mediated Metabolism in Food and Drug Administration-Approved Oral and Intravenous Drugs: 2006-2015.

In recent years, claims of increased involvement of non-cytochrome P450 (non-P450) enzymes in the metabolism of drugs have appeared in the literature. However, no temporal summaries of the contribution of non-P450 enzymes to the metabolism of drugs have been published. Using data from human radiolabeled absorption, distribution, metabolism, and excretion studies available for a set of 125 orall...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:

دوره   شماره 

صفحات  -

تاریخ انتشار 2005