Phosphorodiamidate Morpholino Antisense Oligomers Inhibit Expression of Human Cytochrome P450 3A4 and Alter Selected Drug Metabolism
| Autor: | Patrick L. Iversen, Melissa L. Cate, Chandramallika Ghosh, Vikram Arora |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Paclitaxel
Morpholino CYP3A Morpholines Cell Pharmaceutical Science Substrate Specificity Cytochrome P-450 Enzyme System Coumarins Gene expression medicine Cytochrome P-450 CYP3A Cytochrome P-450 Enzyme Inhibitors Humans Cyclophosphamide Fluorescent Dyes Pharmacology biology Cytochrome P450 Transfection Oligonucleotides Antisense Phosphorus Compounds Molecular biology Growth Inhibitors medicine.anatomical_structure Enzyme inhibitor Hepatocytes biology.protein Fluorescein Caco-2 Cells Cisplatin Drug metabolism |
| Zdroj: | Drug Metabolism and Disposition. 30:757-762 |
| ISSN: | 1521-009X 0090-9556 |
| DOI: | 10.1124/dmd.30.7.757 |
| Popis: | Antisense phosphorodiamidate morpholino oligomers (PMO) inhibit targeted gene expression by preventing ribosomal assembly, thus preventing translation. Inhibition of cytochrome P450 (P450) 3A4 expression was examined in primary human hepatocytes from 11 donors and in Caco-2 cells (stably transfected with CYP3A4 cDNA on an extrachromosomal vector) by evaluating the metabolism of substrate 7-benzyloxy-4-[trifluoromethyl]-coumarin and Western immunoblot analysis. Cellular uptake of PMO was confirmed in both cell systems using fluorescein-labeled PMOs. Three antisense PMO sequences and two control PMO sequences were tested. AVI-4557, a 20-mer PMO with the sequence 5'-CTGGGATGAGAGCCATCACT-3' was selected as the optimal agent. AVI-4557 inhibited expression of CYP3A4 in Caco-2/h3A4 cells by 64% at 24 h following administration of 2.8 microM by an assisted delivery protocol. Inhibition of CYP3A activity was observed in primary human hepatocytes after 24 h exposure to AVI-4557 by an average of 32 +/- 11%. Furthermore, AVI-4557 exposure resulted in a sequence-dependent inhibition of cyclophosphamide-related cytocidal activity and a sequence-dependent induction of paclitaxel-related cytocidal activity in both cell types. Finally, the cytocidal activity of cisplatin was not affected with AVI-4557 treatment in either cell type. These studies indicate AVI-4557 is an effective and specific inhibitor of CYP3A4 expression. |
| Databáze: | OpenAIRE |
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