Skip to main content
SpringerLink
Log in

Is metabolism an important arbiter of anticancer activity of ether lipids? metabolism of SRI 62-834 and hexadecylphosphocholine by [31P]-NMR spectroscopy and comparison of their cytotoxicities with those of their metabolites

  • Original Articles
  • Ether Lipids, Metabolism, Cytotoxicity, Phospholipase, 31p-NMR Spectroscopy
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Summary

Antineoplastic ether lipids have entered phase I clinical trial and, although their mechanism of action remains unclear, it is widely believed that the plasma membrane is the primary cellular drug target. In the present study the hypothesis was tested that metabolism of ether lipids acts as a detoxification process. [31P]-nuclear magnetic resonance (NMR) spectroscopy was used to study the metabolism of the ether lipid SRI 62-834 (SRI) and the phosphate ester hexadecylphosphocholine (HPC) in the presence of both isolated phospholipases C and D and post-mitochondrial rat liver homogenate. Both SRI and HPC were slowly metabolised by phospholipase D to their alkyl phosphates and choline, and the alkyl phosphates were subsequently metabolised by phosphatase to yield the alcohols and inorganic phosphate. These studies failed to detect any metabolism of either SRI or HPC by phospholipase C, and the metabolism of platelet-activating factor (PAF) by this enzyme was not inhibited by the addition of either compound. The cytotoxicity of SRI, the related compound HPC and their metabolites was determined in vitro using three cell lines. Cytotoxicity was measured by analysis of cell growth kinetics, MTT assay and lactate dehydrogenase release. Closely similar results were obtained in the JB1 rat hepatoma cell line, in the non-transformed BL8 rat hepatocyte cell line, and in A549 human lung adenocarcinoma cells. SRI was the most toxic of the compounds analysed, the concentration required to produce 50% toxicity or growth inhibition (IC50) being 6–9 μm. The putative metabolite of SRI, 2,2′-bis(hydroxymethyl)tetrahydrofuran, and the known metabolites [2′-(octadecyloxymethyl)tetrahydrofuran-2′-yl]methyl phosphate and 2-hydroxymethyl-2-octadecyloxymethyltetrahydrofuran exhibited IC50 values of >200, >100 and 40–70 μm, respectively, consistent with metabolic detoxification. HPC was more cytotoxic (IC50, 37 μm) than its phosphate metabolite (IC50, 140 μm), but its toxicity was similar to that of its metabolite hexadecanol (IC50, 28 μm), suggesting that only the former metabolic route leads to detoxification.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BnP:

benzylphosphonate

ET18O CH3 :

2-methyl-1-octadecyl-glycero-3-phosphocholine

HPC:

hexadecylphosphocholine

LDH:

lactate dehydrogenase

MES:

morpholinoethanesulphonate

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide

NMR:

nuclear magnetic resonance

PAF:

platelet-activating factor, 2-acetyl-l-octadecyl-glycero-3-phosphocholine

PBS:

phosphate-buffered saline

SRI 62-834:

SRI, [2′-(octadecyloxymethyl)tetrahydrofuran-2′-yl]methyl 2-[N,N,N-trimethylammonio]ethylphosphate

References

  1. Amouroux R, Chastrette F, Chastrette M (1981) Synthèse de polyéthers polycliques: 1. Stéréosélectivité de l'heterocyclisation d'alcools tétrahydrofurfuryliques γ, δéthyléniques par les sels mercuriques. J Heterocyc Chem 18: 565

    Google Scholar 

  2. Bazill GW, Dexter TM (1989) An antagonist to platelet activating factor counteracts the tumouricidal action of alkyl lysophospholipids. Biochem Pharmacol 38: 374

    Google Scholar 

  3. Bazill GW, Dexter TM (1990) Role of endocytosis in the action of ether lipids in WEH1-3B, HL60 and FDCP-mix A4 cells. Cancer Res 50: 7505

    Google Scholar 

  4. Berdel WE, Bausert WR, Weltzein HU, Modolell ML, Widman KH, Munder PG (1980) The influence of alkyl-lysophospholipids and lysophospholipid-activated macrophages on the development of metastasis of 3-Lewis lung carcinoma. Eur J Cancer 16: 1199

    Google Scholar 

  5. Berdel WE, Bausert WRE, Fink U, Rastetter J, Munder PG (1981) Anti-tumour action of alkyl-lysophospholipids. Anticancer Res 1: 345

    Google Scholar 

  6. Bergmeyer HU, Bernt E (1974) Lactate dehydrogenase. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol II. Academic Press, New York, p 574

    Google Scholar 

  7. Bertz RT (1963) Furtural diacetate (2-furanmethanediol, diacetate). Org Synth Coll IV: 489

    Google Scholar 

  8. Bomalaski JS, Clark MA (1987) The effect ofsn-2 fatty acid substitution on phospholipase C enzyme activities. Biochem J 244: 497

    Google Scholar 

  9. Breiser A, Kim D-J, Fleer EA, Damenz W, Drube A, Berger M, Nagel GA, Eibl H, Unger C (1987) Distribution and metabolism of hexadecylphosphocholine in mice. Lipids 22: 95

    Google Scholar 

  10. Dive C, Watson JV, Workman P (1991) Multiparametric flow cytometry of the modulation of tumor cell membrane permeability by developmental antitumor ether lipid SRI 62-834 in EMT6 mouse mammary tumor and HL-60 human promyelocytic leukemia cells. Cancer Res 51: 799

    Google Scholar 

  11. Eftax DSP, Dunlop AP (1961) New diols in the furan and pyran series. J Org Chem 26: 2106

    Google Scholar 

  12. El-Sayed MY, DeBose CD, Coury LA, Roberts MF (1985) Sensitivity of phospholipase C (Bacillus cereus) activity to phosphatidylcholine structural modifications. Biochim Biophys Acta 837: 325

    Google Scholar 

  13. Fleer EAM, Unger C, Kim D-J, Eibl H (1987) Metabolism of ether phospholipids and analogs in neoplastic cells. Lipids 22: 856

    Google Scholar 

  14. Houlihan WJ, Lee ML, Munder PG, Nemecek GM, Handley DA, Winslow CM, Happy J, Jaeggi C (1987) Antitumour activity of SR 62-834, a cyclic ether analog of ET-18-OCH3. Lipids 22: 884

    Google Scholar 

  15. Lazenby CM, Thompson MG, Hickman JA (1990) Elevation of leukemic cell intracellular calcium by ether lipid SRI62-834. Cancer Res 50: 3327

    Google Scholar 

  16. Manson MM, Legg RF, Watson JV, Green JA, Neal GE (1981) An examination of the relative resistances to aflatoxin B1 and susceptibilities to γ-glutamyl p-phenylene diamine mustard of γ-glutamyl transferase negative and positive cell lines. Carcinogenesis 2: 661

    Google Scholar 

  17. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: applications to proliferation and cytotoxicity assays. J Immunol Methods 65: 55

    Google Scholar 

  18. Noseda A, White JG, Godwin PL, Jerome WG, Modest EJ (1989) Membrane damage in leukemic cells induced by ether and ester lipids: an electron microscopic study. Exp Mol Pathol 50: 69

    Google Scholar 

  19. Okamoto S, Olson AC, Vogler WR, Winton EF (1987) Purging leukemic cells from simulated human remission marrow with alkyllysophospholipids. Blood 69: 1381

    Google Scholar 

  20. Okayasu T, Hoshii K, Seyama K, Ishibashi T, Imai Y (1986) Metabolism of platelet-activating factor in primary cultured adult rat hepatocytes by a new pathway involving phospholipase C and alkyl monooxygenase. Biochim Biophys Acta 876: 58

    Google Scholar 

  21. Perich JW, Johns RB (1988) Di-tert-butylN,N-diethylphosphoramidite. A new phosphitylating agent for the efficient phosphorylation of alcohols. Synthesis 142

  22. Seewald MJ, Olsen RA, Sehgal I, Melder DC, Modest EJ, Powis G (1990) Inhibition of growth factor-dependent inositol phosphate Ca2+ signaling by antitumor ether lipid analogues. Cancer Res 50: 4458

    Google Scholar 

  23. Still WC, Kahn M, Mitra A (1978) Rapid chromatographic technique for preparative separations with moderate resolution. J Org Chem 43: 2923

    Google Scholar 

  24. Twentyman PR, Luscombe M (1987) A study of some variables in a tetrazolium dye (MTT) based assay for cell growth and chemosensitivity. Br J Cancer 56: 279

    Google Scholar 

  25. Überall F, Oberhuber H, Maly K, Zaknun J, Demuth L, Grunicke HH (1991) Hexadecylphosphocholine inhibits inositol phosphate formation and protein kinase C activity. Cancer Res 51: 807

    Google Scholar 

  26. Unger C, Eibl H, Kim DJ, Fleer EA, Koetting J, Bartsch HH, Nagel GA, Pfitzenmaier K (1987) Sensitivity of leukemia cell lines to cytotoxic alkyl-lysophospholipids in relation toO-alkyl cleavage enzyme activities. J Natl Cancer Inst 78: 219

    Google Scholar 

  27. Unger C, Damenz W, Fleer EA, Kim DJ, Breiser A, Hilgard P, Engel J, Nagel G, Eibl H (1989) Hexadecylphosphocholine, a new ether lipid analogue. Studies on the antineoplastic activity in vitro and in vivo. Acta Oncol 28: 213

    Google Scholar 

  28. Vallari DS, Smith ZL, Snyder F (1988) HL-60 cells become resistant towards antitumor ether-linked phospholipids following differentiation into a granulocytic form. Biochem Biophys Res Commun 156: 1

    Google Scholar 

  29. Wilcox RW, Wykle RL, Schmitt JD, Daniel LW (1987) The degradation of platelet-activating factor and related lipids: susceptibility to phospholipases C and D. Lipids 22: 800

    Google Scholar 

  30. Workman P (1991) Antitumour ether lipids: endocytosis as a determinant of cellular sensitivity. Cancer Cells 3: 315

    Google Scholar 

  31. Zheng B, Oishi K, Shoji M, Eibl H, Berdel WE, Hajdu J, Vogler WR, Kuo JF (1990) Inhibition of protein kinase C, (sodium plus potassium)-activated adenosine triphosphatase, and sodium pump by synthetic phospholipid analogues. Cancer Res 50: 3025

    Google Scholar 

Download references

Author information

Author notes

  1. Frances E. Bishop

    Present address: Department of Pharmaceutics, School of Pharmacy, University of Washington, 98 195, Seattle, Washington, USA

  2. Caroline Dive

    Present address: Department of Physiological Sciences, Manchester University, M13 9PT, Manchester, UK

Authors and Affiliations

  1. Cancer Research Campaign Experimental Chemotherapy Group, Pharmaceutical Science Institute, Aston University, B4 7ET, Birmingham, UK

    Frances E. Bishop, Caroline Dive, Sally Freeman & Andreas Gescher

Authors
  1. Frances E. Bishop
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Caroline Dive
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sally Freeman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Andreas Gescher
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by CRC programme grant SP1518 and by an MRC postgraduate fellowship (awarded to F. E. B.). One of the authors (S. E.) is a Lister Institute Fellow and another (C. D.) was supported by the Wolfson Trust

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bishop, F.E., Dive, C., Freeman, S. et al. Is metabolism an important arbiter of anticancer activity of ether lipids? metabolism of SRI 62-834 and hexadecylphosphocholine by [31P]-NMR spectroscopy and comparison of their cytotoxicities with those of their metabolites. Cancer Chemother. Pharmacol. 31, 85–92 (1992). https://doi.org/10.1007/BF00685092

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00685092

Keywords

Search

Navigation