Tandem Benzophenone Amino Pyridines, Potent and Selective Inhibitors of Human Leukotriene C4 Synthase.
Cysteinyl leukotrienes (cys-LTs) are lipid mediators of inflammation. The enzyme catalyzing synthesis of cys-LTs, leukotriene C4 synthase (LTC4S), is considered an important drug target. Here we report the synthesis and characterization of three tandem benzophenone amino pyridines as inhibitors of LTC4S in vitro and in vivo. The inhibitors were characterized in vitro using recombinant human LTC4S, MonoMac 6 cells, and a panel of peripheral human immune cells. In vivo, the compounds were tested in the Zymosan A-induced peritonitis mouse model. The molecules, denoted TK04, TK04a, and TK05, were potent and selective inhibitors of LTC4S with IC50 values of 116, 124, and 95 nM, respectively. Molecular docking revealed binding in a hydrophobic crevice between two enzyme monomers and interaction with two catalytic residues, Arg104 and Arg31. The TK compounds potently inhibited cys-LT biosynthesis in immune cells. In coincubations of platelets and polymorphonuclear leukocytes, inhibition of LTC4S led to shunting of LTA4 toward anti-inflammatory lipoxin A4, which was significantly enhanced by simultaneous inhibition of LTA4H. Finally, we found that TK05 (6 mg⋅kg(-1)⋅body weight) reduces LTE4 levels in peritoneal lavage fluid by 88% and significantly decreases vascular permeability in vivo. Our findings indicate that the TK compounds are valuable experimental tools in eicosanoid research in vitro and in vivo. Their chemical structures may serve as leads for further inhibitor design. Novel drugs depleting cys-LT production could be beneficial for treatment of inflammatory diseases associated with overexpression of LTC4S.
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Thea K
TK
Kleinschmidt
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Martin
M
Haraldsson
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Devaraj
D
Basavarajappa
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Erik
E
Lundeberg
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Madhuranayaki
M
Thulasingam
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Maria
M
Ekoff
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Alexander
A
Fauland
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Christoph
C
Lehmann
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Astrid S
AS
Kahnt
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.).
Lennart
L
Lindbom
Department of Medical Biochemistry and Biophysics (T.K.K., D.B., M.T., A.F., J.Z.H.) and Chemical Biology Consortium Sweden, Science for Life Laboratory Stockholm, Department of Medical Biochemistry and Biophysics (M.H.), and Microvascular Physiology Research Group, Department of Physiology and Pharmacology (E.L., L.L.), Karolinska Institutet, Stockholm Sweden; Clinical Immunology and Allergy Unit, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden (M.E.); and Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt/Main, Germany (C.L., A.S.K.) Jesper.Haeggstrom@ki.se.
Jesper Z
JZ
Haeggström
D000818:Animals; D001577:Benzophenones / Q000737:chemistry*; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology; D002454:Cell Differentiation / Q000187:drug effects; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D004804:Eosinophils / Q000187:drug effects / Q000201:enzymology; D005312:Fetal Blood / Q000166:cytology; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D062105:Molecular Docking Simulation; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology; D011487:Protein Conformation; D011725:Pyridines / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D013379:Substrate Specificity
D001577:Benzophenones; D004791:Enzyme Inhibitors; D011725:Pyridines; C047723:benzophenone; D005982:Glutathione Transferase; C041116:leukotriene-C4 synthase