Abstract
Development of immunomodulatory agents that enhance innate immune responses represents a promising strategy for combating infectious diseases. In the present studies, we screened a series of 71 arylcarboxylic acid hydrazide derivatives for their ability to induce macrophage tumor necrosis factor α (TNF-α) production and identified six such compounds, including one compound previously shown to be a formyl peptide receptor (FPR/FPRL1) agonist. The two most potent compounds [compound 1, nicotinic acid [5-(3-bromophenyl)-2-furyl]methylene-hydrazide; compound 2, 4-fluoro-benzoic acid [5-(3-trifluoromethyl-phenyl)-2-furyl]-methylene-hydrazide] were selected for further analysis. These compounds induced de novo production of TNF-α in a dose- and time-dependent manner in human and murine monocyte/macrophage cell lines and in primary macrophages. These compounds also induced mobilization of intracellular Ca2+, production of reactive oxygen species, and chemotaxis in human and murine phagocytes. Induction of macrophage TNF-α production was pertussis toxin-sensitive, and analysis of the cellular target of these compounds showed that they were FPRL1-specific agonists and that this response was blocked by FPR/FPRL1 and FPRL1-specific antagonists. In addition, pharmacophore modeling showed a high degree of similarity for low-energy conformations of these two compounds to the current pharmacophore model for FPR ligands (
Mol Pharmacol:-1310, 2005). Overall, these compounds represent novel FPRL1 agonists that induce TNF-α, a response distinct from those induced by other known FPR and FPRL1 agonists.
Footnotes
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This work was supported in part by Department of Defense grant W9113M-04-1-0001, National Institutes of Health grants P20-RR020185 and U54-AI065357, National Institutes of Health contract HHSN266200400009C, an equipment grant from the M.J. Murdock Charitable Trust, and the Montana State University Agricultural Experimental Station.
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ABBREVIATIONS: TNF, tumor necrosis factor; LPS, lipopolysaccharide; TLR, Toll-like receptor; FPR, formyl peptide receptor; FPRL1, formyl peptide receptor-like 1; GPCR, G-protein-coupled receptor; ROS, reactive oxygen species; L-012, 8-Amino-5-chloro-7-phenylpyridol[3,4-d]pyridazine-1,4(2H,3H)-dione; ActD, actinomycin D; CHX, cycloheximide; DMSO, dimethyl sulfoxide; fMLF, N-formyl-Met-Leu-Phe; PMA, phorbol 12-myristate 13-acetate; IL, interleukin; KC, keratinocyte-derived chemokine; PTX, pertussis toxin; RBL, rat basophilic leukemia; Go6976, 12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-5H-indolo(2,3-a)pyrrolo(3,4-c)-carbazole; U73122, 1-[6-[[17β-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione; AM, acetoxymethyl ester; DMEM, Dulbecco's modified Eagle's medium; HBSS, Hanks' balanced salt solutions; FBS, fetal bovine serum; NF-κB, nuclear factor κB; ELISA, enzyme-linked immunosorbent assay; RMS, root-mean-square.
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↵ The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.
- Received March 5, 2008.
- Accepted May 5, 2008.
- The American Society for Pharmacology and Experimental Therapeutics
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