title: The 5-lipoxygenase inhibitor RF-22c potently suppresses leukotriene biosynthesis in cellulo and blocks bronchoconstriction and inflammation in vivo.
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.
authors.affiliation: Department of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, University of Jena, Philosophenweg 14, D-07743 Jena, Germany. Electronic address: anja.schaible@gmx.de.
authors.forename: Anja M
authors.initials: AM
authors.lastname: Schaible
authors.affiliation: Department of Experimental Medicine, Second University of Naples, Via Costantinopoli, 16 - I-80138 Naples, Italy; Consorzio Sannio Tech, Via Appia, SNC - 82030 Apollosa (BN), Italy. Electronic address: rosanna.filosa@unina2.it.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Filosa
authors.affiliation: Department of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, University of Jena, Philosophenweg 14, D-07743 Jena, Germany. Electronic address: verena.krauth@uni-jena.de.
authors.forename: Verena
authors.initials: V
authors.lastname: Krauth
authors.affiliation: Institute of Pharmacy/Pharmacognosy, University of Innsbruck, Center for Molecular Biosciences Innsbruck (CMBI), Innrain 80/82, AT-6020 Innsbruck, Austria. Electronic address: Veronika.Temml@uibk.ac.at.
authors.forename: Veronika
authors.initials: V
authors.lastname: Temml
authors.affiliation: Department of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, University of Jena, Philosophenweg 14, D-07743 Jena, Germany. Electronic address: simona.pace@uni-jena.de.
authors.forename: Simona
authors.initials: S
authors.lastname: Pace
authors.affiliation: Department of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, University of Jena, Philosophenweg 14, D-07743 Jena, Germany. Electronic address: ulrike.garscha@uni-jena.de.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Garscha
authors.affiliation: Department of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, University of Jena, Philosophenweg 14, D-07743 Jena, Germany. Electronic address: stefanie.liening@uni-jena.de.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Liening
authors.affiliation: Institute of Transfusion Medicine, University Hospital Jena, D-07743 Jena, Germany. Electronic address: christina.weinigel@med.uni-jena.de.
authors.forename: Christina
authors.initials: C
authors.lastname: Weinigel
authors.affiliation: Institute of Transfusion Medicine, University Hospital Jena, D-07743 Jena, Germany. Electronic address: Silke.Rummler@med.uni-jena.de.
authors.forename: Silke
authors.initials: S
authors.lastname: Rummler
authors.affiliation: Leibniz Institute for Natural Product Research and Infection Biology, Hans-Knöll-Institut, Adolf-Reichwein-Str. 23, D-07745 Jena, Germany. Electronic address: sebastian.schieferdecker@leibniz-hki.de.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Schieferdecker
authors.affiliation: Leibniz Institute for Natural Product Research and Infection Biology, Hans-Knöll-Institut, Adolf-Reichwein-Str. 23, D-07745 Jena, Germany; Department of Biochemical and Chemical Engineering, Technical Biology, Technical University Dortmund, Emil-Figge-Straße 66, D-44227 Dortmund, Germany. Electronic address: markus.nett@leibniz-hki.de.
authors.forename: Markus
authors.initials: M
authors.lastname: Nett
authors.affiliation: Department of Experimental Medicine, Second University of Naples, Via Costantinopoli, 16 - I-80138 Naples, Italy. Electronic address: antonellapeduto@gmail.com.
authors.forename: Antonella
authors.initials: A
authors.lastname: Peduto
authors.affiliation: Department of Experimental Medicine, Second University of Naples, Via Costantinopoli, 16 - I-80138 Naples, Italy. Electronic address: s.collarile@hotmail.it.
authors.forename: Selene
authors.initials: S
authors.lastname: Collarile
authors.affiliation: Department of Experimental Medicine, Second University of Naples, Via Costantinopoli, 16 - I-80138 Naples, Italy. Electronic address: mascuotto1985@gmail.com.
authors.forename: Maria
authors.initials: M
authors.lastname: Scuotto
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano, I-80136 Naples, Italy. Electronic address: roviezzo@cds.unina.it.
authors.forename: Fioretina
authors.initials: F
authors.lastname: Roviezzo
authors.affiliation: Department of Experimental Medicine, Second University of Naples, Via Costantinopoli, 16 - I-80138 Naples, Italy. Electronic address: giuseppe.spaziano@alice.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Spaziano
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano, I-80136 Naples, Italy. Electronic address: mario.derosa@unina2.it.
authors.forename: Mario
authors.initials: M
authors.lastname: de Rosa
authors.affiliation: Institute of Pharmacy/Pharmacognosy, University of Innsbruck, Center for Molecular Biosciences Innsbruck (CMBI), Innrain 80/82, AT-6020 Innsbruck, Austria. Electronic address: hermann.stuppner@uibk.ac.at.
authors.forename: Hermann
authors.initials: H
authors.lastname: Stuppner
authors.affiliation: Institute of Pharmacy/Pharmaceutical Chemistry, University of Innsbruck, Center for Molecular Biosciences Innsbruck (CMBI), Innrain 80/82, AT-6020 Innsbruck, Austria. Electronic address: Daniela.Schuster@uibk.ac.at.
authors.forename: Daniela
authors.initials: D
authors.lastname: Schuster
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via D. Montesano, I-80136 Naples, Italy. Electronic address: bruno.dagostino@unina2.it.
authors.forename: Bruno
authors.initials: B
authors.lastname: D'Agostino
authors.affiliation: Department of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, University of Jena, Philosophenweg 14, D-07743 Jena, Germany. Electronic address: oliver.werz@uni-jena.de.
authors.forename: Oliver
authors.initials: O
authors.lastname: Werz
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D016227:Benzoquinones / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D016084:Bronchoconstriction / Q000187:drug effects* / Q000276:immunology; D002478:Cells, Cultured; D004487:Edema / Q000188:drug therapy / Q000201:enzymology / Q000276:immunology; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D005260:Female; D006801:Humans; D015289:Leukotrienes / Q000096:biosynthesis*; D016859:Lipoxygenase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use; D008807:Mice, Inbred BALB C; D062105:Molecular Docking Simulation; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology / Q000276:immunology
chemical_list: C000619249:4,5-dimethoxy-3-dodecyl-1,2-benzoquinone; D000893:Anti-Inflammatory Agents; D016227:Benzoquinones; D015289:Leukotrienes; D016859:Lipoxygenase Inhibitors; D001094:Arachidonate 5-Lipoxygenase
keywords: 5-Lipoxygenase; Benzoquinones; Inflammation; Leukocytes; Leukotriene