Document Detail

Fluorescence-based assay for the interaction of small molecules with the human renal organic anion transporter 1.
MedLine Citation:
PMID:  10929807     Owner:  NLM     Status:  MEDLINE    
Secretion of small molecules from the systemic blood circulation into urine is one of the physiologically essential functions of the kidney. The human organic anion transporter (hOAT1) is a key component in the renal tubular secretion of negatively charged molecules including a variety of important therapeutics. In some cases, compounds interacting with hOAT1 may induce pharmacokinetic drug-drug interactions or cause nephrotoxicity. We developed a fluorescence-based, 96-well format assay using CHO cells stably expressing hOAT1, which allows for the evaluation of interactions between small molecules and hOAT1. The assay is based on the inhibition of the transport of 6-carboxyfluorescein, a high-affinity hOAT1 substrate (Km = 3.9 microM), which was identified as one of several fluorescent organic anions. The relative inhibition potency of various known hOAT1 substrates determined using the 6-carboxyfluorescein-based inhibition assay correlated well with their Km values, indicating that the fluorescent assay exhibits a proper specificity. This in vitro assay can be employed to evaluate the mechanism of renal clearance of organic anions, to assess potential drug-drug interactions and/or nephrotoxic effects of various therapeutics, and to screen for novel hOAT1 inhibitors that could serve as efficient nephroprotectants.
T Cihlar; E S Ho
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Analytical biochemistry     Volume:  283     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2000 Jul 
Date Detail:
Created Date:  2000-11-06     Completed Date:  2001-01-11     Revised Date:  2005-11-17    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  49-55     Citation Subset:  IM    
Gilead Sciences, Foster City, California 94404, USA.
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MeSH Terms
Anion Transport Proteins
Anions / metabolism
Arylsulfonates / chemistry
Boron Compounds / chemistry
CHO Cells
Carrier Proteins / metabolism*
DNA, Complementary / metabolism
Dose-Response Relationship, Drug
Fluorescein / chemistry
Fluoresceins / chemistry,  metabolism
Fluorescent Dyes / chemistry
Glutarates / pharmacology
Inhibitory Concentration 50
Models, Chemical
Spectrometry, Fluorescence / methods*
Time Factors
Reg. No./Substance:
0/4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; 0/Anion Transport Proteins; 0/Anions; 0/Arylsulfonates; 0/Boron Compounds; 0/Carrier Proteins; 0/DNA, Complementary; 0/Fluoresceins; 0/Fluorescent Dyes; 0/Glutarates; 2321-07-5/Fluorescein; 3301-79-9/6-carboxyfluorescein; 6358-69-6/pyranine; 76823-03-5/4-carboxyfluorescein

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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