Pharmacological profile and efficiency in vivo of diflapolin, the first dual inhibitor of 5-lipoxygenase-activating protein and soluble epoxide hydrolase

  • Sci Rep. 2017 Aug 24;7(1):9398. doi: 10.1038/s41598-017-09795-w.
Ulrike Garscha  1 ,  Erik Romp  2 ,  Simona Pace  2 ,  Antonietta Rossi  3 ,  Veronika Temml  4 ,  Daniela Schuster  4 ,  Stefanie König  2 ,  Jana Gerstmeier  2 ,  Stefanie Liening  2 ,  Markus Werner  2 ,  Heiner Atze  2 ,  Sandra Wittmann  5 ,  Christina Weinigel  6 ,  Silke Rummler  6 ,  Gerhard K Scriba  2 ,  Lidia Sautebin  3 ,  Oliver Werz  2
Affiliations
  • 1. Chair of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, Friedrich-Schiller-University Jena, Philosophenweg 14, D-07743, Jena, Germany. [email protected].
  • 2. Chair of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, Friedrich-Schiller-University Jena, Philosophenweg 14, D-07743, Jena, Germany.
  • 3. Department of Pharmacy, School of Medicine, University of Naples Federico II, 80131, Naples, Italy.
  • 4. Department of Pharmacy / Pharmaceutical Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80-82, A-6020, Innsbruck, Austria.
  • 5. Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, 60438, Frankfurt, Germany.
  • 6. Institute of Transfusion Medicine, University Hospital Jena, 07743, Jena, Germany.
PMID: 28839250 DOI: 10.1038/s41598-017-09795-w
Abstract

Arachidonic acid (AA) is metabolized to diverse bioactive lipid mediators. Whereas the 5-lipoxygenase-activating protein (FLAP) facilitates AA conversion by 5-lipoxygenase (5-LOX) to pro-inflammatory leukotrienes (LTs), the soluble Epoxide Hydrolase (sEH) degrades anti-inflammatory epoxyeicosatrienoic acids (EETs). Accordingly, dual FLAP/sEH inhibition might be advantageous drugs for intervention of inflammation. We present the in vivo pharmacological profile and efficiency of N-[4-(benzothiazol-2-ylmethoxy)-2-methylphenyl]-N'-(3,4-dichlorophenyl)urea (diflapolin) that dually targets FLAP and sEH. Diflapolin inhibited 5-LOX product formation in intact human monocytes and neutrophils with IC50 = 30 and 170 nM, respectively, and suppressed the activity of isolated sEH (IC50 = 20 nM). Characteristic for FLAP inhibitors, diflapolin (I) failed to inhibit isolated 5-LOX, (II) blocked 5-LOX product formation in HEK cells only when 5-LOX/FLAP was co-expressed, (III) lost potency in intact cells when exogenous AA was supplied, and (IV) prevented 5-LOX/FLAP complex assembly in leukocytes. Diflapolin showed target specificity, as other Enzymes related to AA metabolism (i.e., COX1/2, 12/15-LOX, LTA4H, LTC4S, mPGES1, and cPLA2) were not inhibited. In the zymosan-induced mouse peritonitis model, diflapolin impaired vascular permeability, inhibited cysteinyl-LTs and LTB4 formation, and suppressed neutrophil infiltration. Diflapolin is a highly active dual FLAP/sEH inhibitor in vitro and in vivo with target specificity to treat inflammation-related diseases.

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