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Fluorescence-based high-throughput screening assay for drug interactions with UGT1A6
Anne Soikkeli1, Mika Kurkela, Jouni Hirvonen
1Division of Pharmaceutical Technology, University of Helsinki,Viikinkaari 5E, Helsinki, Finland. anne.soikkeli@helsinki.fi
Assay and Drug Development Technologies
|March 29, 2011
Summary
A new fluorescent assay effectively screens UDP-glucuronosyltransferases (UGTs) like UGT1A6, identifying drug metabolism inhibitors and substrates. This high-throughput method accurately measures glucuronidation rates, aiding drug development.
Area of Science:
- Pharmacology
- Biochemistry
- Drug Metabolism
Background:
- UDP-glucuronosyltransferases (UGTs) are crucial for drug pharmacokinetics.
- Studying UGTs requires efficient tools for drug candidate metabolism assessment.
Purpose of the Study:
- Develop a fluorescent high-throughput screening (HTS) assay for UGT1A6.
- Screen for potential UGT1A6 inhibitors and substrates.
Main Methods:
- Assay based on fluorescence increase from 1-naphthol glucuronidation.
- Real-time monitoring of 1-naphthylglucuronide formation.
- Optimized probe concentration, protein amount, buffer, and DMSO for linearity and signal separation.
Main Results:
- Established optimal assay conditions for UGT1A6.
- Demonstrated assay success with known UGT1A6 substrates (4-hydroxyindole, scopoletin).
- Identified several nonsteroidal anti-inflammatory drugs and salicylate derivatives as UGT1A6 inhibitors, showing >50% inhibition at 500 μM.
Conclusions:
- The developed fluorescent HTS assay is a powerful tool for UGT research.
- Assay results correlate well with the reference high-performance liquid chromatography (HPLC) method.
- This assay facilitates efficient screening of drug candidates and inhibitors for UGT1A6 activity.
