Pharmaceutical Research

, Volume 26, Issue 2, pp 449–458

Cellular Phototoxicity Evoked Through the Inhibition of Human ABC Transporter ABCG2 by Cyclin-dependent Kinase Inhibitors In vitro

  • Ran An
  • Yuichiro Hagiya
  • Ai Tamura
  • Shanshan Li
  • Hikaru Saito
  • Daisuke Tokushima
  • Toshihisa Ishikawa
Research Paper

DOI: 10.1007/s11095-008-9738-5

Cite this article as:
An, R., Hagiya, Y., Tamura, A. et al. Pharm Res (2009) 26: 449. doi:10.1007/s11095-008-9738-5

Abstract

Purpose

The physiological importance of the human ATP-binding cassette (ABC) transporter ABCG2 has been recognized with regard to porphyrin-mediated photosensitivity. Functional impairment owing to inhibition of ABCG2 by drugs or its genetic polymorphisms may lead to the disruption of porphyrin homeostasis, which in turn causes cellular toxicity.

Materials and Methods

We evaluated the impact on photosensitivity of the inhibition by cyclin-dependent kinase (CDK) inhibitors of ABCG2 function. For this purpose, we established new methods for photosensitivity assays by using Flp-In-293 cells and plasma membrane vesicles prepared from Sf9 insect cells. With the new methods, we subsequently tested CDK inhibitors, i.e., purvalanol A, WHI-P180, bohemine, roscovitine, and olomoucine.

Results

Among CDK inhibitors tested, purvalanol A was found to be the most potent inhibitor (IC50 = 3.5 μM) for ABCG2-mediated hematoporphyrin transport. At a concentration of 2.5 μM, it evoked the photosensitivity of ABCG2-expressing Flp-In-293 cells treated with pheophorbide a. WHI-P180 moderately inhibited ABCG2 function, exhibiting weak phototoxicity. In contrast, the phototoxicity of bohemine, roscovitine, and olomoucine were minimal in our assay system.

Conclusions

It is suggested that the planar structure is an important factor for interactions with the active site of ABCG2. The present study provides a new approach to studying drug-induced phototoxicity in vitro.

KEY WORDS

ABCG2 (BCRP/MXR)cyclin-dependent kinase inhibitorpheophorbide aphototoxicityporphyrin

Abbreviations

ABC

ATP-binding cassette

BCRP

breast cancer resistance protein

CDK

cyclin-dependent kinase

D-MEM

Dulbecco’s modified Eagle’s medium

EDTA

ethylenediaminetetraacetic acid

EGFR

epidermal growth factor receptor

EGTA

ethyleneglycol-bis(2-aminoethyl)-N,N,N′,N′-tetracetic acid

EKI-785

N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide

FCS

fetal calf serum

HEPES

2-[4-(2-Hydroxyethyl)-1-piperazinyl]ethanesulfonic acid

HOMO

highest occupied molecular orbital

IC50

concentration leading to a 50%-inhibition

Km

Michaelis–Menten constant

MXR

mitoxantrone resistance

MO

molecular orbital

QSAR

quantitative structure-activity relationship

Tris

tris(hydroxymethyl)aminomethane

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Ran An
    • 1
  • Yuichiro Hagiya
    • 1
  • Ai Tamura
    • 1
  • Shanshan Li
    • 1
  • Hikaru Saito
    • 1
  • Daisuke Tokushima
    • 2
  • Toshihisa Ishikawa
    • 1
  1. 1.Department of Biomolecular Engineering, Graduate School of Bioscience and BiotechnologyTokyo Institute of TechnologyYokohamaJapan
  2. 2.NEC CorporationTokyoJapan