PXR mediated elevation of CYP3A4 expression is a costly problem in drug development as well as a clinical problem due to clinically important drug interactions caused by the enzyme induction. CYP3A4 is responsible for the metabolism of more than 50% of the drugs commonly used today. Many of these, as well as other compounds e.g. in herbal medicines can induce transcription of CYP3A4 and thereby enhance the metabolism of other drugs, rendering them ineffective or more toxic. By using an in vitro assay for CYP3A4 induction, tests can be performed on candidate drugs early in development and thereby save time and resources since CYP3A4 inducers are eliminated from further development. A reporter gene assay was constructed by inserting three modules, which includes PXR binding sites isolated from the CYP3A4 sequence, in front of a luciferase gene. This construct was transfected together with PXR into HEK 293 cells. Induction was evoked by adding rifampicin, a known CYP3A4 inducer, to the medium. After lysis of the HEK cells and addition of luciferase substrate, luminescence intensity was recorded as a measure of induction. The construct worked and consistently showed induction by rifampicin, but could be further improved to yield higher sensitivity.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:liu-14804 |
| Date | January 2008 |
| Creators | Nylén, Frank |
| Publisher | Linköpings universitet, Institutionen för fysik, kemi och biologi |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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