Relevance of p-glycoprotein for the enteral absorption of cyclosporin A: In vitro in vivo correlation
Relevance of p-glycoprotein for the enteral absorption of cyclosporin A: In vitro in vivo correlation
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DOI：
10.1111/j.1476-5381.1996.tb15612.x
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发表时间：
1996-08-01
影响因子：
7.3
通讯作者：
Beglinger, C
中科院分区：
文献类型：
--
作者：
Fricker, G;Drewe, J;Beglinger, C
1通过结合人Caco-2细胞的体外实验和健康志愿者的插管研究，研究了环孢菌素A（CyA）与P-糖蛋白在肠道摄取过程中的相互作用。2 CyA摄取到细胞中是不饱和的，仅表现出低的温度敏感性，表明被动扩散。当确定CyA从顶侧到基底侧穿过Caco-2单层的渗透时，总体运输具有高达1 μ M浓度的明显饱和组分。在较高浓度下，渗透增加过成比例。顶侧至基底侧渗透的动力学参数的计算表明，每个过滤器的K-D为0.5 μ l min（-1）的扩散过程，其在基底外侧至顶端方向被活性系统覆盖，K-M为3.8 μ M，J（max）为6.5皮摩尔min（-1）3与从顶侧的渗透相比，当药物从基底侧给药时，CyA渗透显著更高。在长春碱、柔红霉素和非免疫抑制性CyA衍生物的存在下，CyA的顶侧至基底侧转运增加。所有化合物均抑制P-糖蛋白介导的转运过程。在长春碱存在下，CyA的基底外侧至顶端渗透表现出剂量依赖性降低。柔红霉素在Caco-2细胞单层中的渗透率从基底侧到顶侧也高于反之亦然。存在SDZ PSC 833和环孢菌素A时，基底外侧至顶端的渗透减少。4用单克隆抗体C219对Caco-2细胞进行的蛋白质印迹分析证实了所用细胞系统中存在p-糖蛋白。5当测定健康志愿者胃肠道（GI）中CyA的吸收时，血浆AUC的显著降低取决于胃>空肠/回肠>结肠的吸收位置。吸收的减少与胃肠道（胃）P-糖蛋白mRNA的表达显著相关（r=0.994
1 The interaction of cyclosporin A (CyA) with p-glycoprotein during intestinal uptake was investigated by a combination of in vitro experiments with human Caco-2 cells and an intubation study in healthy volunteers.2 CyA uptake into the cells was not saturable and exhibited only a low temperature sensitivity, suggesting passive diffusion. When the permeation of CyA across Caco-2 monolayers from the apical to the basolateral side was determined, overall transport had an apparently saturable component up to a concentration of 1 mu M. At higher concentrations permeation increased over-proportionally. Calculation of the kinetic parameters of apical to basolateral permeation suggested a diffusional process with a K-D of 0.5 mu l min(-1) per filter, which was overlayed by an active system in basolateral to apical direction with a K-M of 3.8 mu M and a J(max) of 6.5 picomol min(-1) per filter.3 CyA permeation was significantly higher when the drug was given from the basolateral side as compared to the permeation from the apical side. Apical to basolateral transport of CyA was increased in the presence of vinblastine, daunomcyin and a non-immunosuppressive CyA-derivative. All compounds inhibit p-glycoprotein-mediated transport processes. Basolateral to apical permeation of CyA showed a dose-dependent decrease in the presence of vinblastine. Permeation of daunomycin across Caco-2 cell monolayers was also higher from the basolateral to the apical side than vice versa. Basolateral to apical permeation was decreased in the presence of SDZ PSC 833 and cyclosporin A.4 Western blot analysis of Caco-2 cells with the monoclonal antibody C219 confirmed the presence of p-glycoprotein in the used cell system.5 When the absorption of CyA in the gastrointestinal (GI)-tract of healthy volunteers was determined, a remarkable decrease of the plasma AUC could be observed dependent on the location of absorption in the rank order stomach >jejunum/ileum > colon. The decrease in absorption exhibited a marked correlation (r=0.994) to the expression of mRNA for p-glycoprotein over the GI-tract (stomach