Method for measuring bile salt export and/or formation activity

Abstract

A method is provided to measure modulation of bile salt export transport and/or formation activity in hepatocyte or stable cell line preparations by test agents including but not limited to drugs, drug candidates, biologicals, food components, herb or plant components, proteins, peptides, DNA, RNA. Furthermore, the method is to determine modulation of bile salt export transport and/or formation activity not only by said test agents, but further their metabolites or biotransformed products formed in situ. The bile salt export transport and/or formation activity modulation includes but not limited to inhibition, induction, activation and/or regulation. The method can be practiced to identify test agents, which have potential to cause liver injury, drug-drug interactions, and/or can be used as therapeutic agents for the treatment of cholestasis, abnormality of bile salt metabolism, liver diseases and cholesterol abnormality.

1 . A kit for assessing a test agent's effect on bile salt export transport and/or formation activity comprising one or more bile salt precursor compounds and one or more hepatocyte preparations, wherein the hepatocyte preparations are used to prepare hepatocyte suspensions for assessing a test agent's effect on bile salt export transport and/or formation activity. 
     
     
         2 . The kit of  claim 1  wherein the hepatocyte preparation comprises primary hepatocytes and/or hepatocytes derived from stable cell lines. 
     
     
         3 . The kit of  claim 1  wherein the hepatocyte preparation comprises hepatocytes derived from human or animal tissue. 
     
     
         4 . The kit of  claim 3  wherein the hepatocyte preparation comprises hepatocytes derived from human liver tissue. 
     
     
         5 . The kit of  claim 3  wherein the hepatocytes preparation comprises hepatocytes derived from mouse, rat, dog, rabbit or monkey liver tissue. 
     
     
         6 . The kit of  claim 1  wherein the hepatocyte preparation comprises hepatocytes derived from stable cell lines. 
     
     
         7 . The kit of  claim 1  wherein the hepatocyte preparation comprises pooled hepatocytes derived from one or more human liver tissue or one or more animal liver tissue selected from mouse, rat, dog, rabbit and monkey. 
     
     
         8 . The kit of  claim 1  wherein the one or more bile salt precursor compounds comprise individually or in combination cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid and/or derivatives thereof. 
     
     
         9 . The kit of  claim 8  wherein the bile acid precursor comprises cholic acid. 
     
     
         10 . The kit of  claim 8  wherein the bile acid precursor comprises chenodeoxycholic acid. 
     
     
         11 . The kit of  claim 1  wherein the one or more bile salt precursor compounds comprise fluorescent, stable isotope, or radiolabeled bile salt precursor compounds selected from cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid and/or derivatives thereof. 
     
     
         12 . A kit for assessing a test agent's effect on bile salt export transport and/or formation activity comprising one or more bile salt precursor compounds. 
     
     
         13 . The kit of  claim 12  wherein the one or more bile salt precursor compounds comprise individually or in combination cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid and/or derivatives thereof. 
     
     
         14 . The kit of  claim 12  wherein the bile acid precursor comprises cholic acid. 
     
     
         15 . The kit of  claim 12  wherein the bile acid precursor comprises chenodeoxycholic acid. 
     
     
         16 . The kit of  claim 12  wherein the one or more bile salt precursor compounds comprise fluorescent, stable isotope, or radiolabeled bile salt precursor compounds selected from cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid and/or derivatives thereof. 
     
     
         20 . (canceled) 
     
     
         21 . A kit for assessing a test agent's effect on bile salt export transport and/or formation activity comprising one or more fluorescent, stable isotope, or radiolabeled bile salt precursor compounds. 
     
     
         22 . The kit of  claim 21  wherein the one or more fluorescent, stable isotope, or radiolabeled bile salt precursor compounds comprise fluorescent, stable isotope, or radiolabeled cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid and/or derivatives thereof. 
     
     
         23 . The kit of  claim 21  wherein the bile acid precursor comprises fluorescent, stable isotope, or radiolabeled cholic acid. 
     
     
         24 . The kit of  claim 21  wherein the bile acid precursor comprises fluorescent, stable isotope, or radiolabeled chenodeoxycholic acid.