Development of novel, 384-well high-throughput assay panels for human drug transporters: drug interaction and safety assessment in support of discovery research.
Transporter proteins are known to play a critical role in affecting the overall absorption, distribution, metabolism, and excretion characteristics of drug candidates. In addition to efflux transporters (P-gp, BCRP, MRP2, etc.) that limit absorption, there has been a renewed interest in influx transporters at the renal (OATs, OCTs) and hepatic (OATPs, BSEP, NTCP, etc.) organ level that can cause significant clinical drug-drug interactions (DDIs). Several of these transporters are also critical for hepatobiliary disposition of bilirubin and bile acid/salts, and their inhibition is directly implicated in hepatic toxicities. Regulatory agencies took action to address transporter-mediated DDI with the goal of ensuring drug safety in the clinic and on the market. To meet regulatory requirements, advanced bioassay technology and automation solutions were implemented for high-throughput transporter screening to provide structure-activity relationship within lead optimization. To enhance capacity, several functional assay formats were miniaturized to 384-well throughput including novel fluorescence-based uptake and efflux inhibition assays using high-content image analysis as well as cell-based radioactive uptake and vesicle-based efflux inhibition assays. This high-throughput capability enabled a paradigm shift from studying transporter-related issues in the development space to identifying and dialing out these concerns early on in discovery for enhanced mechanism-based efficacy while circumventing DDIs and transporter toxicities.
1Department of Leads Discovery and Optimization, Bristol-Myers Squibb, Princeton, NJ, USA.
Huaping
H
Tang
Ding Ren
DR
Shen
Yong-Hae
YH
Han
Yan
Y
Kong
Praveen
P
Balimane
Anthony
A
Marino
Mian
M
Gao
Sophie
S
Wu
Dianlin
D
Xie
Matthew G
MG
Soars
Jonathan C
JC
O'Connell
A David
AD
Rodrigues
Litao
L
Zhang
Mary Ellen
ME
Cvijic
D001692:Biological Transport / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D017277:Drug Approval; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical; D004347:Drug Interactions; D015507:Drugs, Investigational / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D005456:Fluorescent Dyes; D057166:High-Throughput Screening Assays; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship
D015507:Drugs, Investigational; D005456:Fluorescent Dyes; D026901:Membrane Transport Proteins
high-throughput human drug transporter assays