title: The novel benzimidazole derivative BRP-7 inhibits leukotriene biosynthesis in vitro and in vivo by targeting 5-lipoxygenase-activating protein (FLAP).
abstract: OBJECTIVE
  Leukotrienes (LTs) are inflammatory mediators produced via the 5-lipoxygenase (5-LOX) pathway and are linked to diverse disorders, including asthma, allergic rhinitis and cardiovascular diseases. We recently identified the benzimidazole derivative BRP-7 as chemotype for anti-LT agents by virtual screening targeting 5-LOX-activating protein (FLAP). Here, we aimed to reveal the in vitro and in vivo pharmacology of BRP-7 as an inhibitor of LT biosynthesis.
  
  
  METHODS
  We analysed LT formation and performed mechanistic studies in human neutrophils and monocytes, in human whole blood (HWB) and in cell-free assays. The effectiveness of BRP-7 in vivo was evaluated in rat carrageenan-induced pleurisy and mouse zymosan-induced peritonitis.
  
  
  RESULTS
  BRP-7 potently suppressed LT formation in neutrophils and monocytes and this was accompanied by impaired 5-LOX co-localization with FLAP. Neither the cellular viability nor the activity of 5-LOX in cell-free assays was affected by BRP-7, indicating that a functional FLAP is needed for BRP-7 to inhibit LTs, and FLAP bound to BRP-7 linked to a solid matrix. Compared with the FLAP inhibitor MK-886, BRP-7 did not significantly inhibit COX-1 or microsomal prostaglandin E2 synthase-1, implying the selectivity of BRP-7 for FLAP. Finally, BRP-7 was effective in HWB and impaired inflammation in vivo, in rat pleurisy and mouse peritonitis, along with reducing LT levels.
  
  
  CONCLUSIONS
  BRP-7 potently suppresses LT biosynthesis by interacting with FLAP and exhibits anti-inflammatory effectiveness in vivo, with promising potential for further development.
authors.affiliation: Institute of Pharmacy, Friedrich-Schiller-University Jena, Jena, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Pergola
authors.forename: J
authors.initials: J
authors.lastname: Gerstmeier
authors.forename: B
authors.initials: B
authors.lastname: Mönch
authors.forename: B
authors.initials: B
authors.lastname: Çalışkan
authors.forename: S
authors.initials: S
authors.lastname: Luderer
authors.forename: C
authors.initials: C
authors.lastname: Weinigel
authors.forename: D
authors.initials: D
authors.lastname: Barz
authors.forename: J
authors.initials: J
authors.lastname: Maczewsky
authors.forename: S
authors.initials: S
authors.lastname: Pace
authors.forename: A
authors.initials: A
authors.lastname: Rossi
authors.forename: L
authors.initials: L
authors.lastname: Sautebin
authors.forename: E
authors.initials: E
authors.lastname: Banoglu
authors.forename: O
authors.initials: O
authors.lastname: Werz
mesh_terms: D058946:5-Lipoxygenase-Activating Protein Inhibitors / Q000494:pharmacology*; D058927:5-Lipoxygenase-Activating Proteins / Q000378:metabolism; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D001562:Benzimidazoles / Q000494:pharmacology*; D002351:Carrageenan; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D006801:Humans; D020024:Leukotriene Antagonists / Q000494:pharmacology*; D015289:Leukotrienes / Q000096:biosynthesis*; D008297:Male; D051379:Mice; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology; D010538:Peritonitis / Q000139:chemically induced / Q000201:enzymology / Q000517:prevention & control; D010998:Pleurisy / Q000139:chemically induced / Q000201:enzymology / Q000517:prevention & control; D017208:Rats, Wistar; D015054:Zymosan
chemical_list: C000594600:1-(2-chlorobenzyl)-2-(1-(4-isobutylphenyl)ethyl)-1H-benzimidazole; D058946:5-Lipoxygenase-Activating Protein Inhibitors; D058927:5-Lipoxygenase-Activating Proteins; C548592:ALOX5AP protein, human; C548593:Alox5ap protein, mouse; C548594:Alox5ap protein, rat; D000893:Anti-Inflammatory Agents; D001562:Benzimidazoles; D020024:Leukotriene Antagonists; D015289:Leukotrienes; D002351:Carrageenan; D015054:Zymosan; D001094:Arachidonate 5-Lipoxygenase; C583212:ALOX5 protein, human
keywords: 5-lipoxygenase; 5-lipoxygenase activating protein; benzimidazole; inflammation; leukotriene