# US 10,481,149 B2 · Method for measuring bile salt export transport and/or formation activity > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,481,149 B2 - Document ID: US10481149B2 - Status: Granted - Kind: B2 - Application number: 15/675,388 - Filed: 2017-08-11 - Published: 2017-11-30 - Granted: 2019-11-19 - Earliest claimed: 2013-06-14 - Assignees: Biotranex, LLC - Applicants: Biotranex, LLC - Inventors: Kan He - Web page: https://patentgrep.com/patent/US10481149B2 ## CPC - G01N 33/5067: {Liver cells} - G01N 33/60: involving radioactive labelled substances - G01N 33/92: involving lipids, e.g. cholesterol {, lipoproteins, or their receptors (steroid hormones G01N33/743)} - G01N 33/582: {with fluorescent label} - G01N 33/6848: {Methods of protein analysis involving mass spectrometry} - G01N 2500/00: Screening for compounds of potential therapeutic value - G01N 2800/085: Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## 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. ## Claims ### Claim 1 A method for assessing a test agent's effect on bile salt export transport and/or formation activity comprising: (a) separately incubating a hepatocyte suspension preparation and a bile salt precursor with and without a test agent at about 37° C. under conditions allowing bile salt export transport and/or formation for a time sufficient to assess bile salt export transport and/or formation activity of bile salt precursor derived bile salts, (b) post-incubation separating extracellular media and intracellular media of the hepatocyte suspension preparations; and (c) quantifying the bile salt precursor derived bile salts present in the extracellular and/or intracellular media of the post-incubation hepatocyte suspension preparations. ### Claim 2 (depends on 1) The method of claim 1 wherein the hepatocyte suspension preparation comprises freshly prepared or cryopreserved hepatocytes. ### Claim 3 (depends on 1) The method of claim 1 wherein the hepatocyte suspension preparation comprises hepatocytes derived from human or animal tissue. ### Claim 4 (depends on 3) The method of claim 3 wherein the hepatocytes are derived from human liver tissue. ### Claim 5 (depends on 3) The model system of claim 3 wherein the hepatocytes are derived from mouse, rat, dog, rabbit or monkey liver tissue. ### Claim 6 (depends on 1) The model system of claim 1 wherein the hepatocyte suspension preparation comprises hepatocytes derived from stable cell lines. ### Claim 7 (depends on 1) The method of claim 1 wherein the hepatocyte suspension preparation is a pooled hepatocyte suspension preparation. ### Claim 8 (depends on 1) The method of claim 1 wherein the bile salt precursor compound comprises individually or in combination cholic acid, chenodeoxycholic acid, deoxycholic acid, lithocholic acid and/or derivatives thereof. ### Claim 9 (depends on 8) The method of claim 8 wherein the bile acid precursor comprises cholic acid. ### Claim 10 (depends on 8) The model system of claim 8 wherein the bile acid precursor comprises chenodeoxycholic acid. ### Claim 11 (depends on 1) The method of claim 1 wherein the bile salt precursor derived bile salts comprise glycocholic acid, taurocholic acid, glycodeoxy cholic acid, taurodeoxycholic acid, glycolithocholic acid, taurolithocholic acid, glycochenodeoxycholic acid and/or taurochenodeoxycholic acid. ### Claim 12 (depends on 1) The method of claim 1 wherein the test agent is a drug, drug candidate, food component, herb or plant component, amino acid, peptide, protein, oligonucleotide, DNA, or RNA. ### Claim 13 (depends on 12) The method of claim 12 wherein the test agent is a drug or drug candidate. ### Claim 14 (depends on 1) A method for assessing potential treatments for hepatic cholestasis, drug-drug interactions and drug-induced liver injury comprising the method of claim 1. ### Claim 15 A method of generating a report for evaluating the Bile Salts Export Pump (BSEP) inhibition potential of a test agent comprising results of a test agent's effect on bile salt export transport and/or formation activity obtained by: (a) separately incubating a hepatocyte suspension preparation and a bile salt precursor with and without a test agent at about 37° C. under conditions allowing bile salt export transport and/or formation for a time sufficient to assess bile salt export transport and/or formation activity of bile salt precursor derived bile salt, (b) post-incubation separating extracellular media and intracellular media of the hepatocyte suspension preparations; (c) quantifying the bile salt precursor derived bile salts present in the extracellular and/or intracellular media of the post-incubation hepatocyte suspension preparations; and (d) reporting the results of a test agent's effect on bile salt export transport and/or activity as percent inhibition, IC50, and/or Ki. ### Claim 16 (depends on 15) The method of generating a report of claim 15 wherein the test agent is a drug and/or a drug candidate. ### Claim 17 (depends on 15) The method of generating a report of claim 15 further comprising results for at least one reference agent.