The permeation of acamprosate is predominantly caused by paracellular diffusion across Caco-2 cell monolayers: A paracellular modelling approach.
The permeation of acamprosate is predominantly caused by paracellular diffusion across Caco-2 cell monolayers: A paracellular modelling approach.
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阿坎酸的渗透主要是由跨 Caco-2 细胞单层的旁细胞扩散引起的：一种旁细胞建模方法。
DOI：
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发表时间：
2019
影响因子：
4.9
通讯作者：
B. Steffansen
中科院分区：
文献类型：
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作者：
I. Antonescu;Karina Rasmussen;S. Neuhoff;X. Fretté;M. Karlgren;Christel A. S. Bergström;C. Nielsen;B. Steffansen
在药物开发中，估计渗透性限制化合物在人体中的吸收分数（Fa）是重要但具有挑战性的。为了从穿过过滤器生长的Caco-2细胞单层的表观渗透率（Papp）对此类化合物的Fa进行建模，关键在于阐明化合物的肠渗透机制。本研究旨在完善计算渗透性模型，以研究细胞旁和跨细胞途径对生物利用度约为11%的渗透性受限化合物阿坎酸的跨Caco-2单层Papp的相对贡献。测量了阿坎酸和几种细胞旁标志物分子的Papp。这些Papp值用于细化Caco-2单层的系统特异性参数，即细胞旁孔半径、孔容量和电位降。随后，使用从两个公开的渗透性模型收集的数学模型，将改进的参数用作对测试化合物的渗透性（Pmodelled）进行建模的输入。实验数据表明，阿坎酸Papp穿过Caco-2单层是低的，并且在两个运输方向上是相似的。获得的阿坎酸Papp为1.56 ± 0.28 × 10-7 cm·s-1，与细胞旁通透性的分子标记物即甘露醇2.72 ± 0.24 × 10-7 cm·s-1、荧光黄1.80 ± 0.35 × 10-7 cm·s-1和荧光素的Papp相似，2.10 ± 0.28 × 10-7 cm·s-1，低于阿替洛尔的7.32 ± 0.60 × 10-7 cm·s-1（均数± SEM，n = 3-6），而由细胞单层内化的阿坎酸的终点量Qmonolayer低于甘露醇。阿坎酸不影响单层的屏障功能，因为它既不改变三种细胞旁标志物的Papp，也不改变细胞单层的跨上皮电阻（TEER）。所有细胞旁标志物和阿坎酸的Pmodelled由Ppara组分主导，并与实验获得的Papp相匹配。此外，阿坎酸不抑制Caco-2细胞中表达的溶质载体PEPT 1、TAUT、PAT 1、EAAT 1、B 0、+AT/rBAT、OATP 2B 1和ASBT的探针底物的摄取。因此，Pmodeled估计的井Ppara和细胞旁途径似乎是阿坎酸Papp穿过Caco-2单层的主要机制，而由被动扩散或载体介导的替代跨细胞途径被认为仅起不重要的作用。
In drug development, estimating fraction absorbed (Fa) in man for permeability limited compounds in important but challenging. To model Fa of such compounds from apparent permeabilities (Papp) across filter-grown Caco-2 cell monolayers, it is central to elucidate the intestinal permeation mechanism(s) of the compound. The present study aims to refine a computational permeability model in order to investigate the relative contribution of paracellular and transcellular routes to the Papp across Caco-2 monolayers of the permeability limited compound acamprosate having a bioavailability of ~11%. The Papp of acamprosate and of several paracellular marker molecules were measured. These Papp values were used to refine system-specific parameters of the Caco-2 monolayers, i.e. paracellular pore radius, pore capacity and potential drop. The refined parameters were subsequently used as input in modelling the permeability (Pmodelled) of the tested compounds using mathematical models collected from two published permeability models. The experimental data shows that acamprosate Papp across Caco-2 monolayers is low and similar in both transport directions. The obtained acamprosate Papp, 1.56 ± 0.28 × 10-7 cm∙s-1, is similar to the Papp of molecular markers for paracellular permeability namely mannitol, 2.72 ± 0.24 × 10-7 cm∙s-1, lucifer yellow, 1.80 ± 0.35 × 10-7 cm∙s-1 and fluorescein, 2.10 ± 0.28 × 10-7 cm∙s-1 and lower than that of atenolol 7.32 ± 0.60 × 10-7 cm∙s-1 (mean ± SEM, n = 3-6), while the end-point amount of acamprosate internalized by the cell monolayer, Qmonolayer, was lower than that of mannitol. Acamprosate did not influence the barrier function of the monolayers since it neither altered the Papp of the three paracellular markers, nor the transepithelial electrical resistance (TEER) of the cell monolayer. The Pmodelled for all the paracellular markers and acamprosate was dominated by the Ppara component and matched the experimentally obtained Papp. Furthermore, acamprosate did not inhibit the uptake of probe substrates for the solute carriers PEPT1, TAUT, PAT1, EAAT1, B0,+AT/rBAT, OATP2B1 and ASBT expressed in Caco-2 cells. Thus, the Pmodelled estimated well Ppara and the paracellular route appears to be the predominant mechanism for acamprosate Papp across Caco-2 monolayers, while the alternative transcellular routes, mediated by passive diffusion or carriers, are suggested to only play insignificant roles.