Cytochrome P450 3A4 (CYP3A4) Inhibitor Screening Kit (Fluorometric)

Cytochrome P450 3A4 (CYP3A4, EC 1.14.13.157) 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 transformed to more hydrophilic products to facilitate excretion from the body. CYP3A4 is expressed in high levels in the liver and intestines, where it catalyzes oxidation of an extraordinarily wide variety of structurally distinct ligands. More than half of all small molecule drugs commonly used by humans are metabolized by CYP3A4 and inhibition of CYP3A4-mediated metabolism is a common cause of adverse drug/drug and drug/food interactions and toxicity. In addition, for drugs whose pharmacological activity requires metabolism from a pro-drug form, CYP3A4 inhibition can lead to decreased drug efficacy. BioVision’s CYP3A4 Inhibitor Screening Kit enables rapid screening of drugs and other new chemical entities (NCEs) for compound-CYP3A4 interaction in a reliable, high-throughput fluorescence-based assay. The kit provides a yeast microsomal preparation of human CYP3A4 and cytochrome P450 reductase (CPR) enzymes. The assay utilizes a non-fluorescent CYP3A4 substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 535/587 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence. The concentration of fluorogenic substrate used for screening is roughly equivalent to its Km for CYP3A4, facilitating detection of weak competitive inhibitors. The kit contains a complete set of reagents sufficient for performing 200 reactions in a 96-well plate format.

• Detection method : Fluorescence (Ex/Em 535/587 nm) • Species reactivity : Eukaryotes • Applications : • Rapid, high:throughput screening of drugs and novel ligands. • Development of structure:activity relationship (SAR) models to predict CYP3A4 inhibition liability of novel compounds. • Prediction of adverse drug:drug interaction potential and bioavailability for compounds metabolized by CYP3A4.

• Simple, highly sensitive, high-throughput compatible • Rapid screening of CYP3A4 inhibitors or ligands • Kit includes the canonical CYP3A inhibitor ketoconazole and a stable, recombinant human CYP3A4 co-expressed with NADPH Reductase

Cytochrome P450 3A4, CYP3A4, CYP3A, P450 3A inhibitor

none

• CYP3A4 Assay Buffer • Resorufin Standard (5 mM in DMSO) • CYP3A4 Inhibitor (Ketoconazole) • NADPH Generating System (100X) • β-NADP+ Stock (100X) • CYP3A4 Substrate • Recombinant Human CYP3A4

-20°C

gel pack

12 months

Eukaryotes

• Rapid, high-throughput screening of drugs and novel ligands. • Development of structure-activity relationship (SAR) models to predict CYP3A4 inhibition liability of novel compounds. • Prediction of adverse drug-drug interaction potential and bioavailability for compounds metabolized by CYP3A4.

Samples containing drugs, inhibitors or ligands (compounds that can interact and affect CYP3A activity)

Tissue, pathway, proteinase, peptidase, protease ,acrosin, lipoprotein, activator, caspase, trypsin, papain, esterase inhibitors are proteins or receptor ligands or receptor antagonists that bind to an enzyme receptor and decreases its activity. Since blocking an enzyme's activity can kill a pathogen or correct a metabolic imbalance, many drugs are enzyme inhibitors. Not all receptor antagonist that bind to enzymes are inhibitors; enzyme activator ligands or agonists bind to enzymes and increase their enzymatic activity, while enzyme substrates bind and are converted to products in the normal catalytic cycle of the enzyme.