In vitro P-glycoprotein assays to predict the in vivo interactions of P-glycoprotein with drugs in the central nervous system
In vitro P-glycoprotein assays to predict the in vivo interactions of P-glycoprotein with drugs in the central nervous system
复制标题
DOI：
10.1124/dmd.107.017434
复制
发表时间：
2008-02-01
影响因子：
3.9
通讯作者：
de Morais, Sonia M.
中科院分区：
文献类型：
--
作者：
Feng, Bo;Mills, Jessica B.;de Morais, Sonia M.
在3种P-糖蛋白（P-gp）体外试验中检测了31种结构多样的市售中枢神经系统（CNS）活性药物、1种活性代谢产物和7种非CNS活性化合物：使用MDCK、人MDR 1-MDCK和小鼠MDR 1a-MDCK细胞、ATP酶和钙黄绿素AM抑制的transwell试验。此外，在两种体外模型中测量这些化合物的渗透性：平行人工膜渗透试验和MDCK中的顶侧至基底侧表观渗透性。皮下给药后，在P-gp敲除（KO）和野生型（WT）小鼠中测量了同一组化合物在大脑和血浆中的暴露量。使用MDR 1-MDCK与MDCK transwell试验中的比值，确定32种CNS化合物中的1种药物及其代谢产物利培酮和9-羟基利培酮以及7种非CNS药物中的6种具有阳性外排。来自transwell研究的数据与P-gp KO和WT小鼠之间32种CNS化合物的脑-血浆曲线下面积比相关性良好。此外，在MDR 1-MDCK和Mdr 1a-MDCK transwell试验中检测了3300种辉瑞化合物，人MDR 1-MDCK和小鼠Mdr 1a-MDCK细胞中的外排率之间具有良好的相关性（R-2 = 0.92）。渗透性数据显示，32种CNS化合物中的大多数具有中度至高度被动渗透性。这项工作已经证明，在体外转运蛋白测定有助于理解的作用，P-gp介导的外排活性，在确定的处置CNS药物在体内，和transwell试验是一个有价值的体外试验，以评估人类P-gp与化合物的相互作用，以评估脑渗透的新的化学实体，以治疗CNS疾病。
Thirty-one structurally diverse marketed central nervous system (CNS)-active drugs, one active metabolite, and seven non-CNS-active compounds were tested in three P-glycoprotein (P-gp) in vitro assays: transwell assays using MDCK, human MDR1-MDCK, and mouse Mdr1a-MDCK cells, ATPase, and calcein AM inhibition. Additionally, the permeability for these compounds was measured in two in vitro models: parallel artificial membrane permeation assay and apical-to-basolateral apparent permeability in MDCK. The exposure of the same set of compounds in brain and plasma was measured in P-gp knockout (KO) and wild-type (WT) mice after subcutaneous administration. One drug and its metabolite, risperidone and 9-hydroxyrisperidone, of the 32 CNS compounds, and 6 of the 7 non-CNS drugs were determined to have positive efflux using ratio of ratios in MDR1-MDCK versus MDCK transwell assays. Data from transwell studies correlated well with the brain-to-plasma area under the curve ratios between P-gp KO and WT mice for the 32 CNS compounds. In addition, 3300 Pfizer compounds were tested in MDR1-MDCK and Mdr1a-MDCK transwell assays, with a good correlation (R-2 = 0.92) between the efflux ratios in human MDR1-MDCK and mouse Mdr1a-MDCK cells. Permeability data showed that the majority of the 32 CNS compounds have moderate to high passive permeability. This work has demonstrated that in vitro transporter assays help in understanding the role of P-gp-mediated efflux activity in determining the disposition of CNS drugs in vivo, and the transwell assay is a valuable in vitro assay to evaluate human P-gp interaction with compounds for assessing brain penetration of new chemical entities to treat CNS disorders.