title: High throughput screening identifies ATP-competitive inhibitors of the NLRP1 inflammasome.
abstract: Nod-like receptors (NLRs) are cytoplasmic pattern recognition receptors that are promising targets for the development of anti-inflammatory therapeutics. Drug discovery efforts targeting NLRs have been hampered by their inherent tendency to form aggregates making protein generation and the development of screening assays very challenging. Herein we report the results of an HTS screen of NLR family member NLRP1 (NLR family, pyrin domain-containing 1) which was achieved through the large scale generation of recombinant GST-His-Thrombin-NLRP1 protein. The screen led to the identification of a diverse set of ATP competitive inhibitors with micromolar potencies. Activity of these hits was confirmed in a FP binding assay, and two homology models were employed to predict the possible binding mode of the leading series and facilitate further lead-optimization. These results highlight a promising strategy for the identification of inhibitors of NLR family members which are rapidly emerging as key drivers of inflammation in human disease.
authors.affiliation: Pattern Recognition Receptor DPU, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA. Electronic address: philip.a.harris@gsk.com.
authors.forename: Philip A
authors.initials: PA
authors.lastname: Harris
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Chaya
authors.initials: C
authors.lastname: Duraiswami
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Donald T
authors.initials: DT
authors.lastname: Fisher
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: James
authors.initials: J
authors.lastname: Fornwald
authors.affiliation: Pattern Recognition Receptor DPU, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Sandra J
authors.initials: SJ
authors.lastname: Hoffman
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Glenn
authors.initials: G
authors.lastname: Hofmann
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Ruth
authors.initials: R
authors.lastname: Lehr
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Patricia M
authors.initials: PM
authors.lastname: McCormick
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Leng
authors.initials: L
authors.lastname: Nickels
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Schwartz
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Zining
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Platform Technology & Science, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Guofeng
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Pattern Recognition Receptor DPU, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Marquis
authors.affiliation: Pattern Recognition Receptor DPU, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: John
authors.initials: J
authors.lastname: Bertin
authors.affiliation: Pattern Recognition Receptor DPU, GlaxoSmithKline, Collegeville Road, Collegeville, PA 19426, USA.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Gough
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry* / Q000378:metabolism; D051017:Apoptosis Regulatory Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D057166:High-Throughput Screening Assays; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism*; D062105:Molecular Docking Simulation; D000070576:NLR Proteins; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011720:Pyrazoles / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D013329:Structure-Activity Relationship
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D058847:Inflammasomes; D000070576:NLR Proteins; C427903:NLRP1 protein, human; D011720:Pyrazoles; D011993:Recombinant Fusion Proteins; D000255:Adenosine Triphosphate
keywords: Dihydroisoquinolines; Inflammasome; NLRP1; Pyrazolidinediones