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Accumulation of anthracenyl-amino acid topoisomerase I and II inhibitors in drug-sensitive and drug-resistant human ovarian cancer cell lines determined by high-performance liquid chromatography

  • Original Article
  • Anthracenyl-Amino Acid, HPLC Analysis, Drug Uptake, Drug Resistance, Topoisomerase
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Abstract

Anthracenyl amino acid/dipeptide conjugates (AADC) represent novel structures rationally designed for their DNA-binding properties. A high-performance liquid chromatography method is described for simultaneous determination of five compounds that exhibit novel mechanisms of action as topoisomerase I and II inhibitors. The method uses an Apex ODS-2 column and a mobile phase of 0.25M ammonium acetate/trifluoroacetic acid (pH 3) in methanol with gradient elution. Selective detection is achieved by monitoring at 545 nm, with limits of detection ranging between 2 and 4 ng on the column. AADC are recovered from cell sonicates by solid-phase extraction using C2 cartridges, with extraction efficiencies ranging from 84% to 95%. Drug uptake studies were performed with three active compounds in the human ovarian cancer cell line A2780 and its multi-drug-resistant counterpart 2780AD. Marked differences were observed in the pattern of cellular accumulation produced by each compound. NU/ICRF 505 (tyrosine derivative) was taken up most avidly, reaching plateau levels of 4000 pmol/106 cells after 2 h, with no difference being apparent between A2780 and 2780AD. NU/ICRF 510 (arginine derivative) accumulated slowly in A2780, failing to achieve an equilibrium after 4 h, and appeared to be completely excluded from 2780AD. NU/ICRF 500 (serine derivative) was most rapidly taken up by A2780, producing a plateau of 800 pmol/106 cells after only 30 min with approximately 3-fold less accumulation in 2780AD. These results are correlated to the chemosensitivity of the two cell lines to the three compounds.

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Authors and Affiliations

  1. Imperial Cancer Research Fund, Medical Oncology Unit, Western General Hospital, EH4 2XU, Edinburgh, UK

    Jeffrey Cummings, Ian Meikle, Janet S. Macpherson & John F. Smyth

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  1. Jeffrey Cummings
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  2. Ian Meikle
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  3. Janet S. Macpherson
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  4. John F. Smyth
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Cummings, J., Meikle, I., Macpherson, J.S. et al. Accumulation of anthracenyl-amino acid topoisomerase I and II inhibitors in drug-sensitive and drug-resistant human ovarian cancer cell lines determined by high-performance liquid chromatography. Cancer Chemother. Pharmacol. 37, 103–109 (1995). https://doi.org/10.1007/BF00685636

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  • DOI: https://doi.org/10.1007/BF00685636

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