Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay.

Autor:	Hall J; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Brault A; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Vincent F; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Weng S; Pfizer Centers for Therapeutic Innovation (CTI), Boston, Massachusetts, United States of America., Wang H; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Dumlao D; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Aulabaugh A; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Aivazian D; Pfizer Centers for Therapeutic Innovation (CTI), San Diego, California, United States of America., Castro D; Pfizer Centers for Therapeutic Innovation (CTI), San Diego, California, United States of America., Chen M; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Culp J; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Dower K; Inflammation and Immunology, Pfizer, Cambridge, Massachusetts, United States of America., Gardner J; External Research Solutions, Pfizer, Groton, Connecticut, United States of America., Hawrylik S; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Golenbock D; University of Massachusetts Medical School, Worcester, Massachusetts, United States of America., Hepworth D; Medicine Design, Pfizer, Cambridge, Massachusetts, United States of America., Horn M; Pfizer Centers for Therapeutic Innovation (CTI), San Diego, California, United States of America., Jones L; Medicine Design, Pfizer, Cambridge, Massachusetts, United States of America., Jones P; Medicine Design, Pfizer, Cambridge, Massachusetts, United States of America., Latz E; University of Massachusetts Medical School, Worcester, Massachusetts, United States of America.; Institute of Innate Immunity, University Hospitals Bonn, Bonn, Germany., Li J; Medicine Design, Pfizer, Cambridge, Massachusetts, United States of America., Lin LL; Inflammation and Immunology, Pfizer, Cambridge, Massachusetts, United States of America., Lin W; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Lin D; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Lovering F; Medicine Design, Pfizer, Cambridge, Massachusetts, United States of America., Niljanskul N; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Nistler R; Pfizer Centers for Therapeutic Innovation (CTI), Boston, Massachusetts, United States of America., Pierce B; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Plotnikova O; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Schmitt D; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Shanker S; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Smith J; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Snyder W; Pfizer Centers for Therapeutic Innovation (CTI), San Diego, California, United States of America., Subashi T; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Trujillo J; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Tyminski E; Pfizer Centers for Therapeutic Innovation (CTI), Boston, Massachusetts, United States of America., Wang G; Pfizer Centers for Therapeutic Innovation (CTI), Boston, Massachusetts, United States of America., Wong J; Medicine Design, Pfizer, Groton, Connecticut, United States of America., Lefker B; Medicine Design, Pfizer, Cambridge, Massachusetts, United States of America., Dakin L; Medicine Design, Pfizer, Cambridge, Massachusetts, United States of America., Leach K; Pfizer Centers for Therapeutic Innovation (CTI), Boston, Massachusetts, United States of America.
Jazyk:	angličtina
Zdroj:	PloS one [PLoS One] 2017 Sep 21; Vol. 12 (9), pp. e0184843. Date of Electronic Publication: 2017 Sep 21 (Print Publication: 2017).
DOI:	10.1371/journal.pone.0184843
Abstrakt:	Cyclic GMP-AMP synthase (cGAS) initiates the innate immune system in response to cytosolic dsDNA. After binding and activation from dsDNA, cGAS uses ATP and GTP to synthesize 2', 3' -cGAMP (cGAMP), a cyclic dinucleotide second messenger with mixed 2'-5' and 3'-5' phosphodiester bonds. Inappropriate stimulation of cGAS has been implicated in autoimmune disease such as systemic lupus erythematosus, thus inhibition of cGAS may be of therapeutic benefit in some diseases; however, the size and polarity of the cGAS active site makes it a challenging target for the development of conventional substrate-competitive inhibitors. We report here the development of a high affinity (KD = 200 nM) inhibitor from a low affinity fragment hit with supporting biochemical and structural data showing these molecules bind to the cGAS active site. We also report a new high throughput cGAS fluorescence polarization (FP)-based assay to enable the rapid identification and optimization of cGAS inhibitors. This FP assay uses Cy5-labelled cGAMP in combination with a novel high affinity monoclonal antibody that specifically recognizes cGAMP with no cross reactivity to cAMP, cGMP, ATP, or GTP. Given its role in the innate immune response, cGAS is a promising therapeutic target for autoinflammatory disease. Our results demonstrate its druggability, provide a high affinity tool compound, and establish a high throughput assay for the identification of next generation cGAS inhibitors.
Databáze:	MEDLINE
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