title: Identification of selective small molecule inhibitors of the nucleotide-binding oligomerization domain 1 (NOD1) signaling pathway.
abstract: NOD1 is an intracellular pattern recognition receptor that recognizes diaminopimelic acid (DAP), a peptidoglycan component in gram negative bacteria. Upon ligand binding, NOD1 assembles with receptor-interacting protein (RIP)-2 kinase and initiates a signaling cascade leading to the production of pro-inflammatory cytokines. Increased NOD1 signaling has been associated with a variety of inflammatory disorders suggesting that small-molecule inhibitors of this signaling complex may have therapeutic utility. We utilized a cell-based screening approach with extensive selectivity profiling to search for small molecule inhibitors of the NOD1 signaling pathway. Via this process we identified three distinct chemical series, xanthines (SB711), quinazolininones (GSK223) and aminobenzothiazoles (GSK966) that selectively inhibited iE-DAP-stimulated IL-8 release via the NOD1 signaling pathway. All three of the newly identified compound series failed to block IL-8 secretion in cells following stimulation with ligands for TNF receptor, TLR2 or NOD2 and, in addition, none of the compound series directly inhibited RIP2 kinase activity. Our initial exploration of the structure-activity relationship and physicochemical properties of the three series directed our focus to the quinazolininone biarylsulfonamides (GSK223). Further investigation allowed for the identification of significantly more potent analogs with the largest boost in activity achieved by fluoro to chloro replacement on the central aryl ring. These results indicate that the NOD1 signaling pathway, similarly to activation of NOD2, is amenable to modulation by small molecules that do not target RIP2 kinase. These compounds should prove useful tools to investigate the importance of NOD1 activation in various inflammatory processes and have potential clinical utility in diseases driven by hyperactive NOD1 signaling.
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rickard
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Clark A
authors.initials: CA
authors.lastname: Sehon
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Viera
authors.initials: V
authors.lastname: Kasparcova
authors.affiliation: Biological Reagents and Assay Development, Platform Technology Sciences, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Lorena A
authors.initials: LA
authors.lastname: Kallal
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Haile
authors.affiliation: Screening and Compound Profiling, Platform Technology Sciences, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Xin
authors.initials: X
authors.lastname: Zeng
authors.affiliation: Screening and Compound Profiling, Platform Technology Sciences, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Monica N
authors.initials: MN
authors.lastname: Montoute
authors.affiliation: Biological Reagents and Assay Development, Platform Technology Sciences, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Derek D
authors.initials: DD
authors.lastname: Poore
authors.affiliation: Biological Reagents and Assay Development, Platform Technology Sciences, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Hu
authors.initials: H
authors.lastname: Li
authors.affiliation: Screening and Compound Profiling, Platform Technology Sciences, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Zining
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Eidam
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: John G
authors.initials: JG
authors.lastname: Emery
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Marquis
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Gough
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: John
authors.initials: J
authors.lastname: Bertin
mesh_terms: D000818:Animals; D052160:Benzothiazoles / Q000494:pharmacology*; D006801:Humans; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D053474:Nod1 Signaling Adaptor Protein / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D052999:Quinazolinones / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D013329:Structure-Activity Relationship; D014970:Xanthines / Q000494:pharmacology*
chemical_list: D052160:Benzothiazoles; D053474:Nod1 Signaling Adaptor Protein; D052999:Quinazolinones; D014970:Xanthines