Drug failures often occur through drug-drug interactions involving cytochrome P450s, especially CYP3A4. Designing safer drugs faster requires a sensitive, easy to use technology that can be used for a broad range of applications. Promega's new P450-Glo 3A4 Assay with Luciferin-IPA delivers:
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Cell-Based Applications: the best method for measuring CYP3A4 gene induction!
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Excellent Sensitivity: requires very small amounts of starting material- cells, human liver microsomes, recombinant 3A4...
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Selectivity: the most selective luminogenic substrate available.
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Versatile: Luciferin-IPA is compatible with a broad range of inhibition profiles.
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Robust: No fluorescence interference.
Simple and Fast: "Add and read" assay protocols with short incubation times.
Highly Selective CYP3A4 Substrates: Recombinant human P450s were assayed for activity against Luciferin-IPA using the P450-Glo™ method as shown in figure (mean ± SD, n=3).
Sensitive Cell-based Assay- Fresh Human Hepatocytes: Basal cell activity (untreated fresh human hepatocytes) and induced 3A4 activity (fresh human hepatocytes treated for 48hrs with 10µM rifampicin (rif), 500µM phenobarbitol). Cell were incubated with 4µM luciferin-IPA for 60 minutes.
Inhibition Data Correlates well with Conventional Assays: CYP3A4 inhibition by 9 known inhibitors was measured with a conventional CYP3A4 assay (testosterone (TS) 6β-hydroxylase) and with the Luciferin-IPA CYP3A4 assay. IC50s were calculated and compared.