Journal of Molecular Medicine

, Volume 87, Issue 10, pp 981–993

Cellular membranes function as a storage compartment for celecoxib

  • Thorsten J. Maier
  • Susanne Schiffmann
  • Ivonne Wobst
  • Kerstin Birod
  • Carlo Angioni
  • Marika Hoffmann
  • Jakob J. Lopez
  • Clemens Glaubitz
  • Dieter Steinhilber
  • Gerd Geisslinger
  • Sabine Grösch
Original Article

DOI: 10.1007/s00109-009-0506-8

Cite this article as:
Maier, T.J., Schiffmann, S., Wobst, I. et al. J Mol Med (2009) 87: 981. doi:10.1007/s00109-009-0506-8

Abstract

Celecoxib is a selective cyclooxygenase-2-(COX-2)-inhibitor used to treat inflammation and pain and prevents colorectal cancer in patients at high doses by affecting several non-COX-2 proteins. However, celecoxib concentrations appropriate to inhibit proliferation or to induce apoptosis in cell culture (up to 100 µM) clearly exceed those in human plasma (up to 10 µM). Therefore, we speculated that celecoxib might accumulate in human cells, which may facilitate the drug’s interaction with non-COX-2 proteins. Determination of intracellular celecoxib concentrations by liquid chromatography tandem mass spectrometry gave five- to tenfold higher levels as compared to other coxibs (etoricoxib, valdecoxib, lumiracoxib, and rofecoxib) in different tumor cell types, including human HCA-7 and HCT-116 colon carcinoma cells, BL-41 B lymphocytes, Mono Mac 6 monocytes, and in mouse NIH-3T3 non-tumor fibroblasts. This intracellular accumulation of celecoxib was due to an integration of the drug into cellular phospholipid membranes as demonstrated by nuclear Overhauser spectroscopy/nuclear magnetic resonance. Consequently, celecoxib disturbed the plasma membrane integrity of HCT-116 cells and displayed an increased COX-2-inhibitory potency in HCA-7 cells. The use of other coxibs demonstrated that intracellular accumulation is peculiar of celecoxib. Accumulation of celecoxib in human cells may provide a novel molecular basis for the ability of the drug to interact with non-COX-2 targets in vivo despite comparatively low plasma concentrations.

Keywords

NSAIDs Intracellular accumulation Non-COX-2 targets NOESY-NMR Cyclooxygenase Colon cancer 

Abbreviations

BSA

Bovine serum albumin

COX-2

Cyclooxygenase-2

EDTA

Ethylenediaminetetraacetate

ELISA

Enzyme-linked immunoassay

FAP

Familial adenomatous polyposis

FCS

Fetal calf serum

HPLC

High-performance liquid chromatography

HUVEC

Human umbilical vein endothelial cell

LC-MS/MS

Liquid chromatography coupled with tandem mass spectrometry

MAS NOESY

Magic angle spinning assisted nuclear Overhauser spectroscopy

NMR

Nuclear magnetic resonance

NSAID

Nonsteroidal anti-inflammatory drug

PGE2

Prostaglandin E2

POPC

1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine

TMA-DPH

Trimethylammonium diphenylhexatriene

Supplementary material

109_2009_506_MOESM1_ESM.pdf (604 kb)
Figure S1Relative cross-relaxation rates between 1H substrate and POPC nuclei. Following the full matrix rate analysis (see the text for details), normalized substrate to DMPC cross-peak volumes for assigned aromatic protons (see Figure S2) are plotted in the order of increasing distance from the hydrophobic core of the membrane. For all drugs, a high location probability is observed between lipid chain segment C3 and head group segment R: POPC proton assignments are indicated at the bottom (PDF 604 kb)
109_2009_506_MOESM2_ESM.pdf (864 kb)
Figure S21H NMR spectra and assignments of celecoxib, and its analogs, and POPC (PDF 863 kb)
109_2009_506_MOESM3_ESM.pdf (583 kb)
Figure S3Cross-peak intensities between POPC and DMC with rising mixing times. Plotted are the experimentally measured cross-peak intensities (red dots), and relaxation rate curves calculated with the derived cross-relaxation rates (blue lines). The yellow background indicates intensities of cross-peaks between coxib substrate signals and lipid signals. Curves were plotted a with maximum intensities corresponding to the measured values and b each graph scaled to its maximum value (PDF 582 kb)

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Thorsten J. Maier
    • 1
    • 2
  • Susanne Schiffmann
    • 1
  • Ivonne Wobst
    • 1
  • Kerstin Birod
    • 1
  • Carlo Angioni
    • 1
  • Marika Hoffmann
    • 2
  • Jakob J. Lopez
    • 3
  • Clemens Glaubitz
    • 3
  • Dieter Steinhilber
    • 2
  • Gerd Geisslinger
    • 1
  • Sabine Grösch
    • 1
  1. 1.pharmazentrum frankfurt/ZAFES, Institute of Clinical PharmacologyGoethe-UniversityFrankfurt am MainGermany
  2. 2.Institute of Pharmaceutical Chemistry/ZAFESGoethe-UniversityFrankfurt am MainGermany
  3. 3.Centre for Biomolecular Magnetic Resonance and Institute for Biophysical ChemistryGoethe-UniversityFrankfurt am MainGermany

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