Abstract
Purpose
M17055 is under development as a novel loop diuretic for oral administration. To investigate the molecular mechanism of its gastrointestinal absorption, we initially aimed to clarify the mechanism of uptake of M17055 by Caco-2 cells, focusing on possible involvement of OATP-B (SLCO2B1), which is localized in the apical membranes of human intestinal epithelial cells.
Materials and Methods
The uptake of [14C]M17055 by Caco-2 cells cultured on multi-well dishes was measured after cultivation for 14 days. Uptake of [14C]M17055 by HEK293 cells stably expressing OATP-B (HEK293/OATP-B cells) was also examined.
Results
M17055 uptake by Caco-2 cells was saturable, and was inhibited by various organic anions, including other loop diuretics, and several bile acids. Uptake of M17055 by HEK293/OATP-B cells was much higher than that by mock cells. The inhibitory profiles of various organic anions and the estimated K m values for M17055 uptake were similar in Caco-2 and HEK293/OATP-B cells. Moreover, the values of inhibition constants of several inhibitors for M17055 uptake were comparable in the two cell lines.
Conclusion
Our data suggest that OATP-B plays a major role in the uptake of the novel loop diuretic M17055 from apical membranes in Caco-2 cells.
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Abbreviations
- BSP:
-
bromosulfophthalein
- DIDS:
-
5-isothiocyanato-2-[2-(4-isothiocyanato-2-sulfo-phenyl)ethenyl]benzenesulfonic acid
- MCT1:
-
monocarboxylic transporter 1
- PEPT1:
-
oligopeptide transporter 1
- SLC:
-
solute carrier
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Acknowledgements
We thank Dr. Per Artursson (Uppsala University, Sweden) for providing Caco-2 cells. We also thank Ms Lica Ishida for technical assistance. This study was supported in part by a Grant-in-Aid for Scientific Research provided by the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Nishimura, T., Kubo, Y., Kato, Y. et al. Characterization of the Uptake Mechanism for a Novel Loop Diuretic, M17055, in Caco-2 Cells: Involvement of Organic Anion Transporting Polypeptide (OATP)-B. Pharm Res 24, 90–98 (2007). https://doi.org/10.1007/s11095-006-9127-x
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DOI: https://doi.org/10.1007/s11095-006-9127-x