title: Inhibition of JNK and p38 MAPK phosphorylation by 5-(acetylamino)-4-oxo-6-phenyl-2-hexenoic acid methyl ester and 4-phenyl-butenoic acid decreases substance P-induced TNF-α upregulation in macrophages.
abstract: The interactions between the immune and nervous systems play an important role in immune and inflammatory conditions. Substance P (SP), the undecapeptide RPKPQQFFGLM-NH2, is known to upregulate the production of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α. We report here that 5-(acetylamino)-4-oxo-6-phenyl-2-hexenoic acid methyl ester (AOPHA-Me) and 4-phenyl-3-butenoic acid (PBA), two anti-inflammatory compounds developed in our laboratory, reduce SP-stimulated TNF-α expression in RAW 264.7 macrophages. We also show that AOPHA-Me and PBA both inhibit SP-stimulated phosphorylation of JNK and p38 MAPK. Furthermore, molecular modeling studies indicate that both AOPHA-Me and PBA dock at the ATP binding site of apoptosis signal-regulating kinase 1 (ASK1), a member of the MAPKs upstream of both JNK and p38 MAPK, with predicted interaction energies of -7.0 kcal/mol and -5.9 kcal/mol, respectively; this binding overlaps with that of staurosporine, a known inhibitor of ASK1. Taken together, these findings suggest that AOPHA-Me and PBA inhibition of TNF-α expression in SP-stimulated RAW 264.7 macrophages is a consequence of the inhibition of JNK and p38 MAPK phosphorylation. We have previously shown that AOPHA-Me and PBA inhibit the amidative bioactivation of SP, which also would be expected to decrease formation of pro-inflammatory cytokines. It is conceivable that this dual action of inhibiting amidation and MAPK phosphorylation may be of some advantage in enhancing the anti-inflammatory activity of a therapeutic molecule.
authors.affiliation: School of Chemistry and Biochemistry and the Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA 30332, USA.
authors.forename: Jacob D
authors.initials: JD
authors.lastname: Lucrezi
authors.affiliation: Department of Pharmacological Sciences, College of Pharmacy and Health Sciences, Mercer University, Atlanta, GA 30341, USA.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Burns
authors.affiliation: Department of Pharmacological Sciences, College of Pharmacy and Health Sciences, Mercer University, Atlanta, GA 30341, USA.
authors.forename: Diane F
authors.initials: DF
authors.lastname: Matesic
authors.affiliation: School of Chemistry and Biochemistry and the Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA 30332, USA.
authors.forename: Charlie D
authors.initials: CD
authors.lastname: Oldham
authors.affiliation: School of Chemistry and Biochemistry and the Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA 30332, USA. Electronic address: sheldon.may@chemistry.gatech.edu.
authors.forename: Sheldon W
authors.initials: SW
authors.lastname: May
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002208:Caproates / Q000494:pharmacology*; D002460:Cell Line; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology*; D048670:MAP Kinase Kinase 4 / Q000378:metabolism; D048848:MAP Kinase Kinase Kinase 5 / Q000037:antagonists & inhibitors; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D019311:Staurosporine / Q000494:pharmacology; D013373:Substance P / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: C518642:5-(acetylamino)-4-oxo-6-phenyl-2-hexenoic acid methyl ester; D000893:Anti-Inflammatory Agents; D002208:Caproates; D005229:Fatty Acids, Monounsaturated; D014409:Tumor Necrosis Factor-alpha; C064180:4-phenyl-3-butenoic acid; D013373:Substance P; D048051:p38 Mitogen-Activated Protein Kinases; D048848:MAP Kinase Kinase Kinase 5; D048670:MAP Kinase Kinase 4; D019311:Staurosporine
keywords: JNK; RAW 264.7 macrophage; Substance P; TNF-α; p38 MAPK
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