Oncodesign aacr 2018 development of a high throughput in vitro screening pl...
Pjb Probes 2009
1. Discovery of Epigenetic Chemical Probes Peter J. Brown Project Manager, Epigenetics Nov 19 th 2009 SGC Oxford SGC Toronto SGC Stockholm
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6. Histone DNA Lysine Modification Write Read Erase Acetyl (Ac) HAT Bromo HDAC Methyl (Me n ) HMT Royal KDM Chemical Biology of Epigenetics
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10. HMT Assays in production Struc solved by SGC Struc solved by others Assay in Production BIX-01294
11. HAT Assays in production Struc solved by SGC Struc solved by others Assay in Production
12. Assessing HMT Activity Evys Collazo & Raymond C. Trievel et al. (2005) Analytical Biochemistry 342: 86–92
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14. Design of UNC0224 as a Potent G9a Inhibitor BIX-01294 G9a (Thioglo): IC 50 = 0.11 M GLP (Thioglo): IC 50 = 0.062 M Feng Liu & Xin Chen, CICBDD Abdellah Allali-Hassani, SGC UNC0123 G9a (Thioglo): IC 50 = 0.33 M Reduced MW and lipophilicity while maintaining potency Array-based optimization GLP-BIX-01294 complex Adapted from Chang, et al. 2009, Nat. Stru. Mol. Bio., (16), 316 UNC0224 G9a (Thioglo): IC 50 = 0.015 M GLP: IC 50 = 0.020 M, inactive vs SETD7 & SETD8 Selective over a panel of 30 non-Epi targets (except hitting M 2 at 82% inh at 1 M)
15. UNC0224 is a Potent G9a inhibitor BIX-01294 IC50 = 106 nM UNC0224 IC50 = 15 nM
16. Binding Studies ITC was performed in duplicate with average K d values of 130 ± 18 and 23 ± 8 nM for BIX-01294 and UNC0224 respectively. BIX-01294 UNC0224 0.0 0.5 1.0 1.5 -14 -12 -10 -8 -6 -4 -2 0 2 -0.8 -0.6 -0.4 -0.2 0.0 -10 0 10 20 30 40 50 60 70 80 Time (min) µcal/sec Molar Ratio kcal/mole of injectant 0.0 0.5 1.0 1.5 -14 -12 -10 -8 -6 -4 -2 0 2 -0.8 -0.6 -0.4 -0.2 0.0 -10 0 10 20 30 40 50 60 70 80 Time (min) µcal/sec Molar Ratio kcal/mole of injectant 0.0 0.5 1.0 1.5 -8 -6 -4 -2 0 -0.5 -0.4 -0.3 -0.2 -0.1 0.0 -10 0 10 20 30 40 50 60 70 80 Time (min) µcal/sec Molar Ratio kcal/mole of injectant 0.0 0.5 1.0 1.5 -8 -6 -4 -2 0 -0.5 -0.4 -0.3 -0.2 -0.1 0.0 -10 0 10 20 30 40 50 60 70 80 Time (min) µcal/sec Molar Ratio kcal/mole of injectant
17. Co-crystallization of G9a and UNC0224 Crystals diffracted to better than 1.7Å Deposited in PDB (3K5K)
18. BIX-01294 co-crystallized with GLP [3fpd] UNC0224 co-crystallized with G9a [3k5k] H3K9me2 co-crystallized with GLP [2rfi]
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