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Characterization of the Uptake Mechanism for a Novel Loop Diuretic, M17055, in Caco-2 Cells: Involvement of Organic Anion Transporting Polypeptide (OATP)-B

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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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Authors and Affiliations

  1. Division of Pharmaceutical Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan

    Tomohiro Nishimura, Yoshiyuki Kubo, Yukio Kato, Yoshimichi Sai & Akira Tsuji

  2. Drug Metabolism and Pharmacokinetics Laboratory Research Center, Mochida Pharmaceutical Co., Ltd., 722 Jimba-aza-uenohara, Gotemba, Shizuoka, 412-8542, Japan

    Takuo Ogihara

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  1. Tomohiro Nishimura
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  2. Yoshiyuki Kubo
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Correspondence to Akira Tsuji.

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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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