申请人:UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL
公开号:EP2532739A2
公开(公告)日:2012-12-12
A method of screening a candidate compound for susceptibility to biliary excretion by a hepatocyte transport protein. The method includes the steps of providing a culture of hepatocytes comprising a transport protein, the culture having at least one bile canaliculus; exposing a candidate compound to the culture; and determining an amount of candidate compound in the at least one bile canaliculus, the amount of candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion by the transport protein. In some embodiments determining the amount of candidate compound in the bile canaliculus comprises inhibiting expression of the transport protein, measuring the amount of candidate compound in the bile canaliculus and comparing amounts of compound in the canalicules with and without inhibition of the transport protein. A difference in the amount of candidate compound in the canaliculus indicates susceptibility of the candidate compound to biliary excretion by the transport protein. In one embodiment, expression of the transport protein is inhibited through introduction of a RNA having a sequence corresponding to a coding strand of the gene encoding the transport protein into the hepatocyte. Optionally, the culture of hepatocytes is a long-term culture in a sandwich configuration. The method is particularly applicable to the screening of multiple candidate compounds in a single effort.
一种筛选候选化合物是否易受肝细胞转运蛋白胆汁排泄影响的方法。该方法包括以下步骤:提供包含转运蛋白的肝细胞培养物,该培养物具有至少一个胆管;将候选化合物暴露于该培养物;以及确定至少一个胆管中候选化合物的量,至少一个胆管中候选化合物的量表明候选化合物对转运蛋白胆汁排泄的敏感性。在某些实施方案中,确定候选化合物在胆管中的含量包括抑制转运蛋白的表达,测量候选化合物在胆管中的含量,并比较转运蛋白被抑制和未被抑制时胆管中的化合物含量。管腔中候选化合物量的差异表明候选化合物易受转运蛋白的影响而被胆汁排泄。在一个实施方案中,通过将具有与编码转运蛋白基因的编码链相对应序列的 RNA 导入肝细胞来抑制转运蛋白的表达。可选地,肝细胞的培养是在夹层结构中的长期培养。该方法尤其适用于一次性筛选多种候选化合物。