6-Methyl-2,4-Disubstituted Pyridazin-3(<i>2H</i>)-ones: A Novel Class of Small-Molecule Agonists for Formyl Peptide Receptors

Cilibrizzi, Agostino; Quinn, Mark T.; Kirpotina, Liliya N.; Schepetkin, Igor A.; Holderness, Jeff; Ye, Richard D.; Rabiet, Marie‐Josèphe; Biancalani, Claudio; Cesari, Nicoletta; Graziano, Alessia; Vergelli, Claudia; Pieretti, Stefano; Piaz, Vittorio Dal; Giovannoni, Maria Paola

doi:10.1021/jm900592h

Cited by 52 publications

(96 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Small-molecule FPR1 agonists have also been identified in Table 2 and Supplemental Fig. 1. a number of recent studies (Schepetkin et al, 2007;Cilibrizzi et al, 2009;Kirpotina et al, 2010), the most potent of which had a Ca 21 response EC 50 value of 630 nM (Schepetkin et al, 2007). Potent FPR2 agonists have also been reported with Ca 21 response EC 50 values in the 30-40 nM range (Burli et al, 2006;Frohn et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

Selective Agonists and Antagonists of Formylpeptide Receptors: Duplex Flow Cytometry and Mixture-Based Positional Scanning Libraries

et al. 2013

View full text Add to dashboard Cite

The formylpeptide receptor (FPR1) and formylpeptide-like 1 receptor (FPR2) are G protein-coupled receptors that are linked to acute inflammatory responses, malignant glioma stem cell metastasis, and chronic inflammation. Although several Nformyl peptides are known to bind to these receptors, more selective small-molecule, high-affinity ligands are needed for a better understanding of the physiologic roles played by these receptors. High-throughput assays using mixture-based combinatorial libraries represent a unique, highly efficient approach for rapid data acquisition and ligand identification. We report the superiority of this approach in the context of the simultaneous screening of a diverse set of mixture-based small-molecule libraries. We used a single cross-reactive peptide ligand for a duplex flow cytometric screen of FPR1 and FPR2 in colorcoded cell lines. Screening 37 different mixture-based combinatorial libraries totaling more than five million small molecules (contained in 5,261 mixture samples) resulted in seven libraries that significantly inhibited activity at the receptors. Using positional scanning deconvolution, selective high-affinity (low nM K i ) individual compounds were identified from two separate libraries, namely, pyrrolidine bis-diketopiperazine and polyphenyl urea. The most active individual compounds were characterized for their functional activities as agonists or antagonists with the most potent FPR1 agonist and FPR2 antagonist identified to date with an EC 50 of 131 nM (4 nM K i ) and an IC 50 of 81 nM (1 nM K i ), respectively, in intracellular Ca 21 response determinations. Comparative analyses of other previous screening approaches clearly illustrate the efficiency of identifying receptor selective, individual compounds from mixture-based combinatorial libraries.

show abstract

Section: Discussionmentioning

confidence: 99%

Selective Agonists and Antagonists of Formylpeptide Receptors: Duplex Flow Cytometry and Mixture-Based Positional Scanning Libraries

et al. 2013

View full text Add to dashboard Cite

show abstract

“…Additional FPR2-specific agonists, including compound 25 (Cilibrizzi et al, 2009), AG-09/3, AG-09/4, AG-09/5, AG-09/6, AG-09/8, and AG-09/42 and mixed FPR1/FPR2 agonists AG-09/9, AG-09/10 ( Kirpotina et al, 2010) were superimposed onto the template using the FieldAlign program (FieldAlign Version 2.0.1; Cresset Biomolecular Discovery Ltd., Hertfordshire, UK). The molecular structures were imported into FieldAlign in Tripos MOL2 format.…”

Section: Methodsmentioning

confidence: 99%

Gastrin-Releasing Peptide/Neuromedin B Receptor Antagonists PD176252, PD168368, and Related Analogs Are Potent Agonists of Human Formyl-Peptide Receptors

et al. 2010

Self Cite

View full text Add to dashboard Cite

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 Ca 2ϩ 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 ( -(4-nitrophenyl) 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 EC 50 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.

show abstract

“…This may suggest a new strategy for developing novel medicines that could effectively inhibit neutrophil migration, although it would be necessary to investigate the therapeutic effects of FPR agonists in various disease models, especially in more chronic or severe models. Recently, a number of non-peptidyl compounds acting as FPR agonists have been reported, 20,[43][44][45][46] and more potent therapeutic agents than selective antagonists for chemoattractant receptors may develop from these.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of neutrophil migration in mice by mouse formyl peptide receptors 1 and 2 dual agonist: indication of cross-desensitization in vivo

et al. 2010

View full text Add to dashboard Cite

Summary It has been reported that the stimulation of neutrophils with N‐formyl‐Met‐Leu‐Phe (fMLF), an agonist for formyl peptide receptor (Fpr) 1, renders cells unresponsive to other chemoattractants in vitro. This is known as cross‐desensitization, but its functional relevance in neutrophil migration in vivo has not been investigated. Here, we show that precedent stimulation of mouse neutrophils with compound 43, a non‐peptidyl agonist for mouse Fpr1 and Fpr2, rendered the cells unresponsive to a second stimulation with C5a, leukotriene B4, or keratinocyte‐derived cytokine (KC) in calcium mobilization and chemotaxis assays in vitro. The expression of chemokine (C‐X‐C motif) receptor 2 (CXCR2) on the surface of neutrophils was concomitantly diminished by stimulating the cells with the compound. Moreover, oral administration of the compound to mice before they were exposed to lipopolysaccharide (LPS) aerosol resulted in a dose‐dependent reduction in the neutrophil count in bronchoalveolar lavage fluid. The expression of CXCR2 on blood neutrophils was also reduced in the compound‐administered mice. The recipient mice that underwent adoptive transfer of fluorescence‐labelled neutrophils that had been incubated with the compound showed a substantial decrease in neutrophil counts in bronchoalveolar lavage fluid after they were exposed to LPS, when compared with the control mice to which vehicle‐treated neutrophils had been transferred. These results are consistent with the idea that the agonist for Fpr1 and Fpr2 induced cross‐desensitization in neutrophils and attenuated neutrophil migration into the airways. Our results also revealed the unpredicted effect of an Fpr1 and Fpr2 dual agonist, which may act as a functional antagonist for multiple chemoattractant receptors in vivo.

show abstract

6-Methyl-2,4-Disubstituted Pyridazin-3(2H)-ones: A Novel Class of Small-Molecule Agonists for Formyl Peptide Receptors

Cited by 52 publications

References 26 publications

Selective Agonists and Antagonists of Formylpeptide Receptors: Duplex Flow Cytometry and Mixture-Based Positional Scanning Libraries

Selective Agonists and Antagonists of Formylpeptide Receptors: Duplex Flow Cytometry and Mixture-Based Positional Scanning Libraries

Gastrin-Releasing Peptide/Neuromedin B Receptor Antagonists PD176252, PD168368, and Related Analogs Are Potent Agonists of Human Formyl-Peptide Receptors

Inhibition of neutrophil migration in mice by mouse formyl peptide receptors 1 and 2 dual agonist: indication of cross-desensitization in vivo

Contact Info

Product

Resources

About