Cytochrome P450 2D6 (CYP2D6) Inhibitor Screening Kit (Fluorometric) (BN00928)

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BN00928
  • Cytochrome P450 2D6 (CYP2D6) Inhibitor Screening Kit (Fluorometric) (BN00928)
  • Cytochrome P450 2D6 (CYP2D6) Inhibitor Screening Kit (Fluorometric) (BN00928)
  • Cytochrome P450 2D6 (CYP2D6) Inhibitor Screening Kit (Fluorometric) (BN00928)
€792

Description

ELISA Kit Technical ManualMSDS

Cytochrome P450 2D6 (CYP2D6) Inhibitor Screening Kit (Fluorometric)

Cytochrome P450 2D6 (CYP2D6, EC 1.14.14.1) is a member of the cytochrome P450 monooxidase (CYP) family of microsomal xenobiotic metabolism enzymes. CYPs are membrane-bound hemeproteins responsible for Phase I biotransformation reactions, in which lipophilic drugs and other xenobiotic compounds are converted to more hydrophilic products to facilitate excretion from the body. CYP2D6 catalyzes oxidation of lipophilic bases with an aromatic ring and a nitrogen atom and is highly expressed in liver and brain tissue. The enzyme is responsible for metabolism of nearly 25% of all small molecule drugs commonly used by humans, particularly psychiatric drugs such as antidepressants, antipsychotics and stimulants. The CYP2D6 gene is highly polymorphic in the human population, with resulting CYP2D6 activity ranging from complete metabolic deficiency to ultra-rapid metabolism depending upon allelic variation and gene copy number. Due to this wide phenotypic variability, CYP2D6 is frequently implicated in drug toxicity and clinical drug/drug interactions. In addition, for drugs whose pharmacological activity requires metabolism from a pro-drug form, CYP2D6 inhibition or allelic deficiency can lead to decreased drug efficacy. Assay Genie's CYP2D6 Inhibitor Screening Kit enables rapid screening of drugs and other new chemical entities (NCEs) for compound-CYP2D6 interaction in a reliable, high-throughput fluorescence-based assay. The kit provides a yeast microsomal preparation of human CYP2D6 and cytochrome P450 reductase (CPR) enzymes. The assay utilizes a non-fluorescent CYP2D6-selective substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 390/468 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence. The kit contains a complete set of reagents sufficient for performing 200 reactions in a 96-well plate format.

Figure: (a) AHMC standard curve. One mole of AHMC corresponds to the metabolism of one mole of CYP2D6 substrate. (b) Reaction kinetics of recombinant human CYP2D6 enzyme at 37°C in the presence and absence of the indicated CYP2D6 inhibitors (the solvent control reaction contained a final concentration of 0.5% acetonitrile). (c) Dose-response curves for various CYP2D6 ligands of differing structural and mechanistic classes: the canonical competitive CYP2D6 inhibitor quinidine, dextromethorphan, a prototypical CYP2D6 substrate and the antidepressants fluoxetine and paroxetine (a mechanism-based, irreversible CYP2D6 inhibitor). For dose-response curves, percent activity was calculated for each concentration of inhibitor by comparison to activity of reactions containing no inhibitor. For each CYP2D6 inhibitor, IC 50 values were derived by 4-parameter logistic curve fitting with points representing the mean ± SEM of at least three replicate experiments. Assays were performed according to the kit protocol.

Key Information Description

Product SKU

BN00928

Size

200 assays

Detection Method

Fluorescence (Ex/Em 390/468 nm)

Species Reactivity

Eukaryotes

Applications

  • Rapid, high-throughput screening of drugs and novel ligands.
  • Development of structure-activity relationship (SAR) models to predict CYP2D6 inhibition liability of novel compounds.
  • Prediction of adverse drug-drug interaction potential and bioavailability for compounds metabolized by CYP2D6.

Features and Benefits

  • Simple, highly sensitive, high-throughput compatible
  • Rapid screening of CYP2D6 inhibitors or ligands
  • Kit includes the canonical CYP2D6 inhibitor Quinidine and a stable, recombinant human CYP2D6 co-expressed with NADPH Reductase

Kit Components

  • CYP2D6 Assay Buffer
  • AHMC Standard
  • CYP2D6 Inhibitor (Quinidine)
  • NADPH Generating System (100X)
  • Beta-NADP+ Stock (100X)
  • CYP2D6 Substrate
  • Recombinant Human CYP2D6

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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