Cytochrome P450 2C19 (CYP2C19) Inhibitor Screening Kit (Fluorometric) (BN01065)

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BN01065
  • Cytochrome P450 2C19 (CYP2C19) Inhibitor Screening Kit (Fluorometric) (BN01065)
  • Cytochrome P450 2C19 (CYP2C19) Inhibitor Screening Kit (Fluorometric) (BN01065)
  • Cytochrome P450 2C19 (CYP2C19) Inhibitor Screening Kit (Fluorometric) (BN01065)
€511

Description

ELISA Kit Technical ManualMSDS

Cytochrome P450 2C19 (CYP2C19) Inhibitor Screening Kit (Fluorometric)

Cytochrome P450 2C19 (CYP2C19, 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. CYP2C19 is primarily expressed in liver and intestinal tissue and catalyzes oxidation of neutral or weakly basic lipophilic molecules with 2-3 hydrogen bond donor/acceptor sites. Isoforms of the CYP2C subfamily are responsible for metabolism of nearly 20% of all small molecule drugs commonly used by humans. Polymorphisms in the human CYP2C19 gene have been implicated in clinical drug/drug interactions involving widely-prescribed drugs, including proton pump inhibitors, antiplatelet agents and anticonvulsants. In addition, for drugs whose pharmacological activity requires metabolism from a pro-drug form (such as the antithrombotic drug clopidogrel), CYP2C19 inhibition or allelic deficiency can lead to decreased drug efficacy. Assay Genie's CYP2C19 Inhibitor Screening Kit enables rapid screening of drugs and other new chemical entities (NCEs) for compound-CYP2C19 interaction in a reliable, high-throughput fluorescence-based assay. The kit provides a yeast microsomal preparation of human CYP2C19 and cytochrome P450 reductase (CPR) enzymes. The assay utilizes a non-fluorescent CYP2C19 substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 406/468 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence. The kit contains a complete set of reagents sufficient for performing 100 reactions in a 96-well plate format.

Figure: (a) 3-cyano-7-hydroxycoumarin (3-CHC) standard curve. One mole of 3-CHC corresponds to the metabolism of one mole of CYP2C19 substrate. (b) Reaction kinetics of recombinant human CYP2C19 enzyme at 37°C in the presence and absence of the indicated CYP2C19 inhibitors (the solvent control reaction contained a final concentration of 0.5% acetonitrile). (c) Dose-response curves for various CYP2C19 ligands of differing structural and mechanistic classes: the CYP2C19-selective competitive inhibitor (+)-N- 3-benzylnirvanol, the irreversible CYP2C19 inhibitor ticlopidine, the antidepressants tranylcypromine and fluvoxamine and the proton pump inhibitor lansoprazole. For dose-response curves, percent activity was calculated for each concentration of inhibitor by comparison to activity of reactions containing no inhibitor. For each CYP2C19 inhibitor, IC 50 values were derived by 4-parameter logistic curve fitting with each point representing the mean ± SEM of at least three replicates. Assays were performed according to the kit protocol.

Key Information Description

Product SKU

BN01065

Size

100 assays

Detection Method

Fluorescence (Ex/Em 406/468 nm)

Species Reactivity

Eukaryotes

Applications

  • Rapid, high-throughput screening of drugs and novel ligands.
  • Development of structure-activity relationship (SAR) models to predict CYP2C19 inhibition liability of novel compounds.
  • Prediction of adverse drug-drug interaction potential and bioavailability for compounds metabolized by CYP2C19.

Features and Benefits

  • Simple, highly sensitive, high-throughput compatible
  • Rapid screening of CYP2C19 inhibitors or ligands
  • Kit includes the CYP2C19 inhibitor Ticlopidine and a stable, recombinant human CYP2C19 co-expressed with NADPH Reductase

Kit Components

  • CYP2C19 Assay Buffer
  • 3-CHC Standard
  • CYP2C19 Inhibitor (Ticlopidine)
  • NADPH Generating System (100X)
  • Beta-NADP+ Stock (100X)
  • CYP2C19 Substrate
  • Recombinant Human CYP2C19

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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