High-content screening of clinically tested anticancer drugs identifies novel inhibitors of human MRP1 (ABCC1)

Pharmacol Res. 2017 May;119:313-326. doi: 10.1016/j.phrs.2017.02.024. Epub 2017 Feb 28.


Multidrug resistance protein 1 (MRP1/ABCC1), an integral transmembrane efflux transporter, belongs to the ATP-binding cassette (ABC) protein superfamily. MRP1 governs the absorption and disposition of a wide variety of endogenous and xenobiotic substrates including various drugs across organs and physiological barriers. Additionally, its overexpression has been implicated in multidrug resistance in chemotherapy of multiple cancers. Here, we describe the development of a high content imaging-based screening assay for MRP1 activity. This live cell-based automated microscopy assay is very robust and allows simultaneous detection of cell permeable, non-toxic and potent inhibitors. The validity of the assay was demonstrated by profiling a library of 386 anti-cancer compounds, which are under clinical trials, for interactions with MRP1. The assay identified 12 potent inhibitors including two known MRP1 inhibitors, cyclosporine A and rapamycin. On the other hand, MRP1-inhibitory activity of tipifarnib, AZD1208, deforolimus, everolimus, temsirolimus, HS-173, YM201636, ESI-09, TAK-733, and CX-6258 has not been previously reported. Inhibition of MRP1 activity was further validated using flow cytometry and confocal microscopy for the respective detection of calcein and doxorubicin in MRP1-overexpressing cells. Among the identified compounds, tipifarnib, AZD1208, rapamycin, deforolimus, everolimus, TAK-733, and temsirolimus resensitized MRP1-overexpressing H69AR cells towards vincristine, a cytotoxic chemotherapeutic agent, by 2-6-fold. Using purified HEK293 membrane vesicles overexpressing MRP1, MRP2, MRP3, and MRP4, we also demonstrated that the identified compounds exert differential and selective response on the uptake of estradiol glucuronide, an endogenous MRP substrate. In summary, we demonstrated the effectiveness of the high content imaging-based high-throughput assay for profiling compound interaction with MRP1.

Keywords: ABC transporter; AZD1208 (PubChem CID: 58423153); Anti-cancer agent; CX-6258 (PubChem CID: 44545852); Cyclosporin A (PubChem CID: 5284373); Deforolimus (PubChem CID: 11520894); Drug disposition; Drug profiling; Drug-transporter interactions; ESI-09 (PubChem CID: 6077765); Everolimus (PubChem CID: 6442177); HS-173 (PubChem CID: 52936849); High content imaging; MRP1; TAK-733 (PubChem CID: 24963252); Tipifarnib (PubChem CID: 159324); YM201636 (PubChem CID: 9956222).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Cell Line
  • Drug Evaluation, Preclinical / methods*
  • Drug Resistance, Multiple / drug effects
  • HEK293 Cells
  • High-Throughput Screening Assays / methods*
  • Humans
  • Multidrug Resistance-Associated Proteins / antagonists & inhibitors*
  • Multidrug Resistance-Associated Proteins / metabolism


  • Antineoplastic Agents
  • Multidrug Resistance-Associated Proteins
  • multidrug resistance-associated protein 1