The 5-lipoxygenase inhibitor RF-22c potently suppresses leukotriene biosynthesis in cellulo and blocks bronchoconstriction and inflammation in vivo

Biochem Pharmacol. 2016 Jul 15:112:60-71. doi: 10.1016/j.bcp.2016.04.019. Epub 2016 May 5.

Abstract

5-Lipoxygenase (5-LO) catalyzes the first two steps in leukotriene (LT) biosynthesis. Because LTs play pivotal roles in allergy and inflammation, 5-LO represents a valuable target for anti-inflammatory drugs. Here, we investigated the molecular mechanism, the pharmacological profile, and the in vivo effectiveness of the novel 1,2-benzoquinone-featured 5-LO inhibitor RF-22c. Compound RF-22c potently inhibited 5-LO product synthesis in neutrophils and monocytes (IC50⩾22nM) and in cell-free assays (IC50⩾140nM) without affecting 12/15-LOs, cyclooxygenase (COX)-1/2, or arachidonic acid release, in a specific and reversible manner, supported by molecular docking data. Antioxidant or iron-chelating properties were not evident for RF-22c and 5-LO-regulatory cofactors like Ca(2+) mobilization, ERK-1/2 activation, and 5-LO nuclear membrane translocation and interaction with 5-LO-activating protein (FLAP) were unaffected. RF-22c (0.1mg/kg; i.p.) impaired (I) bronchoconstriction in ovalbumin-sensitized mice challenged with acetylcholine, (II) exudate formation in carrageenan-induced paw edema, and (III) zymosan-induced leukocyte infiltration in air pouches. Taken together, RF-22c is a highly selective and potent 5-LO inhibitor in intact human leukocytes with pronounced effectiveness in different models of inflammation that warrants further preclinical analysis of this agent as anti-inflammatory drug.

Keywords: 5-Lipoxygenase; Benzoquinones; Inflammation; Leukocytes; Leukotriene.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / administration & dosage
  • Anti-Inflammatory Agents / pharmacology*
  • Anti-Inflammatory Agents / therapeutic use
  • Arachidonate 5-Lipoxygenase / metabolism*
  • Benzoquinones / administration & dosage
  • Benzoquinones / pharmacology*
  • Benzoquinones / therapeutic use
  • Blood Platelets / drug effects
  • Blood Platelets / enzymology
  • Blood Platelets / immunology
  • Bronchoconstriction / drug effects*
  • Bronchoconstriction / immunology
  • Cells, Cultured
  • Edema / drug therapy
  • Edema / enzymology
  • Edema / immunology
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Female
  • Humans
  • Leukotrienes / biosynthesis*
  • Lipoxygenase Inhibitors / administration & dosage
  • Lipoxygenase Inhibitors / pharmacology*
  • Lipoxygenase Inhibitors / therapeutic use
  • Mice, Inbred BALB C
  • Molecular Docking Simulation
  • Monocytes / drug effects
  • Monocytes / enzymology
  • Monocytes / immunology
  • Neutrophils / drug effects
  • Neutrophils / enzymology
  • Neutrophils / immunology

Substances

  • 4,5-dimethoxy-3-dodecyl-1,2-benzoquinone
  • Anti-Inflammatory Agents
  • Benzoquinones
  • Leukotrienes
  • Lipoxygenase Inhibitors
  • Arachidonate 5-Lipoxygenase