6NAO

Discovery of a high affinity inhibitor of cGAS

Replaces:  5V8N

Summary for 6NAO

• Entry DOI: 10.2210/pdb6nao/pdb
• Related: 5V8H 5V8J 5V8O
• Descriptor: CYCLIC GMP-AMP SYNTHASE, ZINC ION, (1R,2S)-2-[(7-hydroxy-5-phenylpyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]cyclohexane-1-carboxylic acid, ... (4 entities in total)
• Functional Keywords: cgas, sting, cgamp, transferase, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 2
• Total formula weight: 85701.42

Authors

Hall, J. (deposition date: 2018-12-06, release date: 2018-12-19, Last modification date: 2024-03-13)

Primary citation

Hall, J.,Brault, A.,Vincent, F.,Weng, S.,Wang, H.,Dumlao, D.,Aulabaugh, A.,Aivazian, D.,Castro, D.,Chen, M.,Culp, J.,Dower, K.,Gardner, J.,Hawrylik, S.,Golenbock, D.,Hepworth, D.,Horn, M.,Jones, L.,Jones, P.,Latz, E.,Li, J.,Lin, L.L.,Lin, W.,Lin, D.,Lovering, F.,Niljanskul, N.,Nistler, R.,Pierce, B.,Plotnikova, O.,Schmitt, D.,Shanker, S.,Smith, J.,Snyder, W.,Subashi, T.,Trujillo, J.,Tyminski, E.,Wang, G.,Wong, J.,Lefker, B.,Dakin, L.,Leach, K.
Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay.
PLoS ONE, 12:e0184843-, 2017

PubMed Abstract: 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.

PubMed: 28934246
DOI: 10.1371/journal.pone.0184843

Experimental method

X-RAY DIFFRACTION (3.23 Å)