Method of screening candidate compounds for susceptibility to biliary excretion

Abstract

A method of screening a candidate compound for susceptibility to biliary excretion. The method includes the steps of providing a culture of hepatocytes, 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. 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.

1 . A method of screening a candidate compound for susceptibility to biliary excretion, the method comprising the steps of:
 (a) providing a culture of hepatocytes, the culture of hepatocytes comprising at least one bile canaliculus;   (b) exposing a candidate compound to the culture; and   (c) determining an amount of candidate compound in the at least one bile canaliculus, the amount of the candidate compound in the at least one bile canaliculus indicating the susceptibility of the candidate compound to biliary excretion.   
   
   
       2 . The method of  claim 1 , wherein the hepatocytes are isolated from a source selected from the group consisting of rat, human, monkey, ape, cat, dog, pig, hog, cattle, oxen, sheep, horses, turkeys, chickens, ducks and geese. 
   
   
       3 . The method of  claim 1 , wherein the culture of hepatocytes further comprises a long-term culture of hepatocytes. 
   
   
       4 . The method of  claim 1 , wherein the culture of hepatocytes further comprises a canalicular network. 
   
   
       5 . The method of  claim 1 , wherein the culture of hepatocytes is further characterized as having a configuration selected from the group consisting of clusters of hepatocytes, aggregates of hepatocytes, at least one layer of hepatocytes, and combinations thereof. 
   
   
       6 . The method of  claim 5 , wherein the hepatocytes are embedded in a matrix. 
   
   
       7 . The method of  claim 5 , wherein the culture of hepatocytes further comprises a sandwich culture of hepatocytes, the sandwich culture comprising at least one layer of hepatocytes and at least one bile canaliculus with the at least one layer of hepatocytes. 
   
   
       8 . The method of  claim 7 , wherein the sandwich culture of hepatocytes further comprises a long-term sandwich culture of hepatocytes. 
   
   
       9 . The method of  claim 7 , wherein the at least one layer of hepatocytes is sandwiched between two layers of matrix. 
   
   
       10 . The method of  claim 9 , wherein the matrix is selected from the group consisting of a biological matrix medium, a synthetic matrix medium, and combinations thereof. 
   
   
       11 . The method of  claim 10 , wherein the biological matrix medium is selected from the group consisting of collagens, laminins, basement membrane-derived complexes, derivatives thereof and combinations thereof. 
   
   
       12 . The method of  claim 1 , wherein the amount of the candidate compound in the at least one bile canaliculus is determined by calculating a biliary clearance value for the culture, the calculated biliary clearance value indicating the susceptibility of the candidate compound to biliary excretion. 
   
   
       13 - 64 . (canceled)