Characterization of the efflux transporter(s) responsible for restricting intestinal mucosa permeation of an acyloxyalkoxy-based cyclic prodrug of the opioid peptide DADLE.
Characterization of the efflux transporter(s) responsible for restricting intestinal mucosa permeation of an acyloxyalkoxy-based cyclic prodrug of the opioid peptide DADLE.
复制标题
负责限制阿片肽 DADLE 的基于酰氧基烷氧基的环状前药肠粘膜渗透的外排转运蛋白的表征。
DOI：
10.1023/a:1016144530146
复制
发表时间：
2002
影响因子：
3.7
通讯作者：
Borchardt,RonaldT
中科院分区：
文献类型：
--
作者：
Tang,Fuxing;Borchardt,RonaldT
目的。目的：阐明阿片肽DADLE(AD)的外排转运体(S)对阿片肽DADLE(AD)环状前体药物透过Caco-2细胞单层的限制作用。用Caco-2细胞、Madin-Darby犬肾野生型II细胞(MDCK-WT)、转人MDR1基因的MDCK细胞(MDCK-MDR1)和转人MRP2基因的MDCK细胞(MDCK-MRP2)研究AD的细胞渗透特性。这些细胞以单分子层的形式生长在微孔膜上。用高效液相色谱检测AD在供体侧的消失和在受体侧的出现。用已知的P-糖蛋白特异性抑制剂GF120918测定AD对P-糖蛋白(P-gp)的底物活性。用已知的MRP2和P-gp抑制剂环孢素A测定AD对MRP2的底物活性。在Caco-2细胞中，测得的AD流量在基底(BL)和根尖(AP)方向的表观渗透系数(Papp)与AP-to-BL方向的流量(Papp BL-to-AP/Papp AP-to-BL)之比为99。在2μM GF120918或25μM环孢素A存在下，PAPP BL与AP/PAPP AP与BL的比值降至11。在MDCK-WT、MDCK-MDR1和MDCK-MRP2细胞中，AD的PAPP BL与AP/PAPP AP与BL的比值分别为4.7、10和5.8。GF120918(2μM)和环孢素A(25μM)的混合物可使MDCK-WT、MDCK-MDR1和MDCK-MRP2细胞中AD的Papp BL/AP/Papp AP/BL比值分别降低到1.2、1.8和2.3。这些数据表明AD是一种比MRP2更好的P-gp底物，这种环状前药在Caco-2细胞和肠粘膜中的限制渗透可能主要是由于它对P-gp的底物活性。
Purpose. To elucidate the efflux transporter(s) responsible for restricting the permeation of an acyloxyalkoxy-based cyclic prodrug of the opioid peptide DADLE (AD) through Caco-2 cell monolayers.Methods. The cellular permeation characteristics of AD were investigated using Caco-2 cells, Madin-Darby canine kidney wild-type II cells (MDCK-WT), MDCK cells transfected with the human MDR1 gene (MDCK-MDR1), and MDCK cells transfected with the human MRP2 gene (MDCK-MRP2). These cells were grown as monolayers onto microporous membranes. The disappearance of AD from the donor side and its appearance on the receiver side were monitored by high-performance liquid chromatography. The substrate activity of AD for P-glycoprotein (P-gp) was determined using GF120918, a known P-gp specific inhibitor. The substrate activity of AD for MRP2 was determined by using cyclosporin A, a known MRP2 and P-gp inhibitor.Results. In Caco-2 cells, the ratio of the apparent permeability coefficients (Papp) of AD flux measured in the basolateral (BL) to apical (AP) direction vs. the flux in the AP-to-BL direction (Papp BL-to-AP/Papp AP-to-BL) was 99. In the presence of 2 μM GF120918 or 25 μM cyclosporin A, the Papp BL-to-AP/Papp AP-to-BLratio was decreased to 11. In MDCK-WT, MDCK-MDR1, and MDCK-MRP2 cells, the Papp BL-to-AP/Papp AP-to-BLratios of AD were 4.7, 10, and 5.8, respectively. A mixture of GF120918 (2 μM) and cyclosporin A (25 μM) decreased the Papp BL-to-AP/Papp AP-to-BLratios of AD in MDCK-WT, MDCK-MDR1, and MDCK-MRP2 cells to 1.2, 1.8, and 2.3, respectively.Conclusions. These data suggest that AD is a much better substrate for P-gp than MRP2 and that the restricted permeation of this cyclic prodrug in Caco-2 cells and in the intestinal mucosa probably is due primarily to its substrate activity for P-gp.