Assessment of Bile Salt Export Pump (BSEP) Inhibition in Membrane Vesicles Using Radioactive and LC/MS-Based Detection Methods.
The bile salt export pump (BSEP, ABCB11) belongs to the ATP-binding-cassette superfamily of transporters and is predominately found in the liver. BSEP is an efflux transporter that plays a critical role in the secretion of bile salts into the bile. Inhibition of BSEP function by drugs can result in the buildup of bile salts in the liver and eventually leads to cholestasis and drug-induced liver injury (DILI). DILI is a major cause of withdrawal of drugs from the pharmaceutical market and accounts for >50% of acute liver failures. Therefore, early detection of BSEP inhibition by drugs can help to mitigate the possibility of BSEP-associated liver injury. This unit describes two assays that investigate the relationship between drug interference with BSEP function and liver injury using membrane vesicles prepared from Hi5 insect cells transfected with human BSEP. Comprehensive protocols for assessing BSEP inhibition in a 384-well format using radiolabeled and liquid chromatography/mass spectrometry (LC/MS)-based detection methods are described. © 2017 by John Wiley & Sons, Inc.
Hit Discovery and Optimization, Pfizer Inc, Groton, Connecticut.
Lisa D
LD
Marroquin
Primary Pharmacology Group, Pfizer Inc, Groton, Connecticut.
Paul D
PD
Bonin
Hit Discovery and Optimization, Pfizer Inc, Groton, Connecticut.
Julie
J
Keefer
Hit Discovery and Optimization, Pfizer Inc, Groton, Connecticut.
Thomas
T
Schroeter
D000074020:ATP Binding Cassette Transporter, Subfamily B, Member 11 / Q000037:antagonists & inhibitors*; D000818:Animals; D002460:Cell Line; D056486:Chemical and Drug Induced Liver Injury; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D020128:Inhibitory Concentration 50; D007313:Insecta; D057230:Limit of Detection; D011874:Radiometry / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D018675:Toxicity Tests
C093491:ABCB11 protein, human; D000074020:ATP Binding Cassette Transporter, Subfamily B, Member 11
ATP-binding cassette transporter; BSEP; DILI; bile salt export pump; cholestasis; drug induced liver injury