TY - JOUR
T1 - Characterization of the uptake mechanism for a novel loop diuretic, M17055, in Caco-2 cells
T2 - Involvement of organic anion transporting polypeptide (OATP)-B
AU - Nishimura, Tomohiro
AU - Kubo, Yoshiyuki
AU - Kato, Yukio
AU - Sai, Yoshimichi
AU - Ogihara, Takuo
AU - Tsuji, Akira
N1 - Funding Information:
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.
PY - 2007/1
Y1 - 2007/1
N2 - 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.
AB - 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.
KW - Caco-2
KW - Intestinal absorption
KW - Loop diuretics
KW - Membrane transport
KW - OATP-B
KW - Transporter
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U2 - 10.1007/s11095-006-9127-x
DO - 10.1007/s11095-006-9127-x
M3 - Article
C2 - 17103337
AN - SCOPUS:33845460145
SN - 0724-8741
VL - 24
SP - 90
EP - 98
JO - Pharmaceutical Research
JF - Pharmaceutical Research
IS - 1
ER -