Biogenic synthesis, purification, and chemical characterization of anti-inflammatory resolvins derived from docosapentaenoic acid (DPAn-6)

J Biol Chem. 2009 May 29;284(22):14744-59. doi: 10.1074/jbc.M809014200. Epub 2009 Mar 26.

Abstract

Enzymatically oxygenated derivatives of the omega-3 fatty acids cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) and cis-5,8,11,14,17-eicosapentaenoic acid, known as resolvins, have potent inflammation resolution activity (Serhan, C. N., Clish, C. B., Brannon, J., Colgan, S. P., Chiang, N., and Gronert, K. (2000) J. Exp. Med. 192, 1197-1204; Hong, S., Gronert, K., Devchand, P. R., Moussignac, R., and Serhan, C. N. (2003) J. Biol. Chem. 278, 14677-14687). Our objective was to determine whether similar derivatives are enzymatically synthesized from other C-22 fatty acids and whether these molecules possess inflammation resolution properties. The reaction of DHA, DPAn-3, and DPAn-6 with 5-, 12-, and 15-lipoxygenases produced oxylipins, which were identified and characterized by liquid chromatography coupled with tandem mass-spectrometry. DPAn-6 and DPAn-3 proved to be good substrates for 15-lipoxygenase. 15-Lipoxygenase proved to be the most efficient enzyme of the three tested for conversion of long chain polyunsaturated fatty acids to corresponding oxylipins. Since DPAn-6 is a major component of Martek DHA-S oil, we focused our attention on reaction products obtained from the DPAn-6 and 15-lipoxygenase reaction. (17S)-hydroxy-DPAn-6 and (10,17S)-dihydroxy-DPAn-6 were the main products of this reaction. These compounds were purified by preparatory high performance liquid chromatography techniques and further characterized by NMR, UV spectrophotometry, and tandem mass spectrometry. We tested both compounds in two animal models of acute inflammation and demonstrated that both compounds are potent anti-inflammatory agents that are active on local intravenous as well as oral administration. These oxygenated DPAn-6 compounds can thus be categorized as a new class of DPAn-6-derived resolvins.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / chemistry*
  • Anti-Inflammatory Agents / isolation & purification*
  • Anti-Inflammatory Agents / pharmacology
  • Anti-Inflammatory Agents / therapeutic use
  • Arachidonate 15-Lipoxygenase / metabolism
  • Cell Movement / drug effects
  • Chromatography, Liquid
  • Docosahexaenoic Acids / chemistry*
  • Dose-Response Relationship, Drug
  • Edema / drug therapy
  • Fatty Acids, Unsaturated / biosynthesis
  • Fatty Acids, Unsaturated / chemistry*
  • Fatty Acids, Unsaturated / isolation & purification*
  • Fatty Acids, Unsaturated / pharmacology
  • Glycine max / enzymology
  • Granulocytes / drug effects
  • Isomerism
  • Kinetics
  • Leukocytes / cytology
  • Leukocytes / drug effects
  • Macrophages / drug effects
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Mice
  • Rats
  • Solubility / drug effects
  • Substrate Specificity / drug effects
  • Sus scrofa

Substances

  • Anti-Inflammatory Agents
  • Fatty Acids, Unsaturated
  • Docosahexaenoic Acids
  • Arachidonate 15-Lipoxygenase
  • docosapentaenoic acid