Abstract
N-Formyl peptide receptors (FPRs) are G protein-coupled receptors (GPCRs) involved in host defense and sensing cellular dysfunction. Thus, FPRs represent important therapeutic targets. In the present studies, we screened 32 ligands (agonists and antagonists) of unrelated GPCRs for their ability to induce intracellular Ca2+ mobilization in human neutrophils and HL-60 cells transfected with human FPR1, FPR2, or FPR3. Screening of these compounds demonstrated that antagonists of gastrin-releasing peptide/neuromedin B receptors (BB1/BB2) PD168368 [(S)-a-methyl-a-[[[(4-nitrophenyl)amino]carbonyl]amino]-N-[[1-(2-pyridinyl) cyclohexyl]methyl]-1H-indole-3-propanamide] and PD176252 [(S)-N-[[1-(5-methoxy-2-pyridinyl)cyclohexyl]methyl]-a-methyl-a-[[-(4-nitrophenyl)amino]carbonyl]amino-1H-indole-3-propanamide] were potent mixed FPR1/FPR2 agonists, with nanomolar EC50 values. Cholecystokinin-1 receptor agonist A-71623 [Boc-Trp-Lys(ε-N-2-methylphenylaminocarbonyl)-Asp-(N-methyl)-Phe-NH2] was also a mixed FPR1/FPR2 agonist, but with a micromolar EC50. Screening of 56 Trp- and Phe-based PD176252/PD168368 analogs and 41 related nonpeptide/nonpeptoid analogs revealed 22 additional FPR agonists. Most were potent mixed FPR1/FPR2/FPR3 agonists with nanomolar EC50 values for FPR2, making them among the most potent nonpeptide FPR2 agonists reported to date. In addition, these agonists were also potent chemoattractants for murine and human neutrophils and activated reactive oxygen species production in human neutrophils. Molecular modeling of the selected agonists using field point methods allowed us to modify our previously reported pharmacophore model for the FPR2 ligand binding site. This model suggests the existence of three hydrophobic/aromatic subpockets and several binding poses of FPR2 agonists in the transmembrane region of this receptor. These studies demonstrate that FPR agonists could include ligands of unrelated GPCR and that analysis of such compounds can enhance our understanding of pharmacological effects of these ligands.
Footnotes
↵
The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.This work was supported in part by the National Institutes of Health National Center for Research Resources [Grant P20-RR020185]; by the National Institutes of Health [Contract HHSN266200400009C]; the M. J. Murdock Charitable Trust; and the Montana State University Agricultural Experimental Station.
Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.
doi:10.1124/mol.110.068288.
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ABBREVIATIONS:
- fMLF
- N-formyl-methionyl-leucyl-phenyalanine
- FPR
- formyl peptide receptor
- GPCR
- G protein-coupled receptor
- SAR
- structure-activity relationship
- ROS
- reactive oxygen species
- L-012
- 8-amino-5-chloro-7-phenylpyridol[3,4-d]pyridazine-1,4(2H,3H)-dione
- DMSO
- dimethyl sulfoxide
- WKYMVm
- Trp-Lys-Tyr-Met-Val-d-Met
- HBSS
- Hanks' balanced salt solution
- HBSS+
- Hanks' balanced salt solution containing 1.3 mM CaCl2 and 1.0 mM MgSO4
- AM
- acetoxymethyl ester
- MPO
- myeloperoxidase
- CCK
- cholecystokinin
- TM
- transmembrane
- PD168368
- (S)-a-methyl-a-[[[(4-nitrophenyl)amino]carbonyl]amino]-N-[[1-(2-pyridinyl) cyclohexyl]methyl]-1H-indole-3-propanamide
- PD176252
- (S)-N-[[1-(5-methoxy-2-pyridinyl)cyclohexyl]methyl]-a-methyl-a-[[-(4-nitrophenyl)amino]carbonyl]amino-1H-indole-3-propanamide
- A-71623
- Boc-Trp-Lys(ε-N-2-methylphenylaminocarbonyl)-Asp-(N-methyl)-Phe-NH2
- NNC 63-0532
- (8-naphthalen-1-ylmethyl-4-oxo-1-phenyl-1,3,8-triaza-spiro(4.5)-dec-3-yl)acetic acid methyl ester
- SR 27897
- 1-((2-(4-(2-chlorophenyl)thiazol-2-yl)aminocarbonyl)indolyl)acetic acid
- YM 022
- 1-(2,3-dihydro-1-(2′-methylphenacyl)-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl)-3-(3-methylphenyl)urea
- LY 288513
- 1-(4-bromophenylaminocarbonyl)-4,5-diphenyl-3-pyrazolidinone
- BML-190
- indomethacin morpholinylamide
- AM 630
- iodopravadoline
- GW 405833
- 1-(2,3-dichlorobenzoyl)-5-methoxy-2-methyl-3-[2-(4-morpholinyl)ethyl]-1H-indole
- S 25585
- 1-benzoyl-2-[[trans-4-[[[[2-nitro-4-trifluoromethyl)phenyl]sulfonyl]amino]methyl]cyclohexyl]carbonyl]hydrazine
- SR 49059
- relcovaptan
- AC 55541
- (2E)-2-[1-(3-bromo-phenyl)ethylidene] α-(benzoylamino)-3,4-dihydro-4-oxo-1-phthalazineacetic acid hydrazide
- JTE 013
- 1-[1,3-dimethyl-4-(2-methylethyl)-1H-pyrazolo[3,4-b]pyridin-6-yl]-4-(3,5-dichloro-4-pyridinyl)-semicarbazide
- RF9
- adamantylcarbonyl-arginyl-phenylalaninamide
- SR 142948
- 2-[[[5-(2,6-dimethoxyphenyl)-1-[4- [[[3-(dimethylamino)propyl]methylamino]carbonyl]-2-(1-methylethyl)phenyl]-1H-pyrazol-3-yl]carbonyl]amino]-tricyclo[3.3.1.13,7]decane-2-carboxylic acid
- FR 139317
- N-(N-(N-((hexahydro-1H-azepin-1-yl)carbonyl)-l-leucyl)-1-methyl-d-tryptophyl)-3-(2-pyridinyl)-d-alanine
- ONO 1078
- pranlukast
- L-692,585
- 3-[[(2R)-2-hydroxypropyl]amino]-3-methyl-N-[(3R)-2,3,4,5-tetrahydro-2-oxo-1-[[2′-(1H-tetrazol-5-yl)[1,1′-biphenyl]-4-yl]methyl]-1H-1-benzazepin-3-yl]-butanamide
- T 98475
- 7-[(2,6-difluorophenyl)methyl]-4,7-dihydro-2-[4-[(2-methyl-1-oxopropyl)amino]phenyl]-3-[[methyl(phenylmethyl)amino]methyl]-4-oxo-thieno[2,3-b]pyridine-5-carboxylic acid 1-methylethyl ester
- L-371,257
- 1-(1-(4-((N-acetyl-4-piperidinyl)oxy)-2-methoxybenzoyl)piperidin-4-yl)-4H-3,1-benzoxazin-2(1H)-one
- FK 888
- N(2)-(4-hydroxy-1-(1-methyl-1H-indol-3-yl)carbonyl-l-prolyl)-N-methyl-N-phenylmethyl-3-(2-naphthyl)-l-alaninamide
- SDZ NKT 343
- 2-nitrophenylcarbamoyl-(S)-prolyl-(S)-3-(2-naphthyl)alanyl-N-benzyl-N-methylamide
- L-732,138
- N-acetyl-l-tryptophan 3,5-bis(trifluoromethyl)benzyl ester
- WEB 2086
- apafant
- L-161,982
- N-[[4′-[[3-butyl-1,5-dihydro-5-oxo-1-[2-(trifluoromethyl)phenyl]-4H-1,2,4-triazol-4-yl]methyl][1,1′-biphenyl]-2-yl]sulfonyl]-3-methyl-2-thiophenecarboxamide
- L-054,264
- 2-((spiro(1H-indene-1,4′-piperidin)-1′-ylcarbonyl)amino)-N-(3-aminomethyl-1-cyclohexylmethyl)-3-(1H-indol-3-yl)propanamide
- BIM 187
- 1-de(5-oxo-l-proline)-2-de-l-valine-3-d-phenylalanine-10-l-leucine-11-l-leucinamide-ranatensin
- BIM 189
- 1-de(5-oxo-l-proline)-2-de-l-valine-3-d-phenylalanine-10-l-leucine-11-(4-chloro-l-phenylalaninamide)-ranatensin
- BIM 23042
- d-Nal-Cys-Tyr-d-Trp-Lys-Val-Cys-Nal-NH2
- BIM 23127
- d-Nal-Cys-Tyr-d-Trp-Orn-Val-Cys-Nal-NH2
- ICI 216,140
- N-isobutyryl-His-Trp-Ala-Val-d-Ala-His-Leu-NHMe
- PD165929
- 2-[3-(2, 6-diisopropyl-phenyl)-ureido]3-(1H-indol-3-yl)-2-methyl-N-(1-pyridin-2-yl-cyclohexylmethyl)-proprionate
- Frohn-11
- 1-((5-methoxyindol-2-yl)carbonyl)-3-(2-ethylbenzimidazol-1-yl)(3R)pyrrolidine
- Bürli-25
- N-(4-bromophenyl)-N-(1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-4-yl)urea
- Cilibrizzi-14x
- N-(4-bromophenyl)-2-[5-(4-methoxybenzyl)-3-methyl-6-oxo-6H-pyridazin-1-yl]-acetamide
- NCGC00168126–01
- N-(4-(5-(3-(furan-2-ylmethyl)-4-oxo-1,2,3,4-tetrahydroquinazolin-2-yl)-2-methoxybenzyloxy)phenyl acetamide.
- Received August 18, 2010.
- Accepted October 13, 2010.
- Copyright © 2011 The American Society for Pharmacology and Experimental Therapeutics
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Gastrin-Releasing Peptide/Neuromedin B Receptor Antagonists PD176252, PD168368, and Related Analogs Are Potent Agonists of Human Formyl-Peptide Receptors
