In vitro and in vivo identification of ABCB1 as an efflux transporter of bosutinib

J Hematol Oncol. 2015 Jul 7:8:81. doi: 10.1186/s13045-015-0179-4.

Abstract

Background: Bosutinib is a recently approved ABL inhibitor. In spite of the well-documented effectiveness of BCR-ABL inhibitors in treating chronic myeloid leukemia, development of resistance is a continuous clinical challenge. Transporters that facilitate drug uptake and efflux have been proposed as one potential source of resistance to tyrosine kinase inhibitor treatment. Our aim was to determine which carriers are responsible for bosutinib transport.

Methods: K562S cells overexpressing the drug transporters ABCB1, ABCG2, and SLC22A1 were generated, characterized and used in proliferation assay and intracellular uptake and retention assay (IUR). In vivo experiments were performed in nude mice injected with K562S, K562DOX cells (overexpressing ABCB1), and K562DOX silenced for ABCB1 (K562DOX/sh P-GP).

Results: The IUR assay using C-14 bosutinib showed that only ABCB1 was responsible for active bosutinib transport. K562DOX cells showed the lowest intracellular level of bosutinib, while K562DOX cells treated with the ABCB1 inhibitor verapamil showed intracellular bosutinib levels comparable with parental K562S. Proliferation assays demonstrated that K562DOX are resistant to bosutinib treatment while verapamil is able to restore the sensitivity to the drug. Nude mice injected with K562DOX and treated with bosutinib showed very limited response and quickly relapsed after stopping treatment while K562S as well as K562DOX/sh P-GP remained tumor-free.

Conclusions: Our data suggest that the analysis of ABCB1 expression levels might help determine treatment options for patients exhibiting resistance to bosutinib.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B / drug effects
  • Aniline Compounds / administration & dosage
  • Aniline Compounds / metabolism
  • Aniline Compounds / therapeutic use*
  • Animals
  • Cell Line, Tumor
  • Cell Proliferation
  • Female
  • Humans
  • In Vitro Techniques
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / drug therapy*
  • Mice
  • Mice, Nude
  • Microscopy, Confocal
  • Nitriles / administration & dosage
  • Nitriles / metabolism
  • Nitriles / therapeutic use*
  • Quinolines / administration & dosage
  • Quinolines / metabolism
  • Quinolines / therapeutic use*
  • Transfection

Substances

  • ABCB1 protein, human
  • ATP Binding Cassette Transporter, Subfamily B
  • Aniline Compounds
  • Nitriles
  • Quinolines
  • bosutinib