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A public-private partnership that supports the discovery of new medicines through open access research.
A-395 A Chemical Probe For EEDThis probe and its inactive control are available from Sigma (A-395 and A-395N) and Cayman (A-395). For any inquiries please contact proberequests@thesgc.org.group newOverview A-395 A-395N (negative control) A collaboration between Abbvie and the SGC has resulted in the discovery of A-395, the first potent and selective chemical probe for EED. The in vitro activity of A-395 includes potent binding to EED with Ki = 0.4 nM, inhibits the PRC2 complex with IC50 = 34 nM for methylation of H3K27 and greater than 100-fold selectivity over other histone methyltransferases and non-epigenetic targets. In cellular assays, A-395 inhibits the PRC2 complex (thus inhibiting the formation of H3K27me3) with IC50 = 90 nM (RD rhabdoid tumor cell line; 3 days). A closely related compound, A-395N, exhibits no activity in the biochemical and cellular assays, and is an ideal control compound for cellular studies. in vitro potency A-395 potently inhibited the formation of H3K27me3 (via antagonizing EED in the trimeric PRC2 complex (EZH2:EED:SUZ12)) with IC50 = 34 ± 2 nM (Hill Slope = 0.7) Properties A-395 A-395N (negative control) Physical and chemical properties for A-395 Molecular weight 486.2 Molecular formula C26H35FN4O2S IUPAC name 1-(5-fluoro-bicyclo[4.3.0]nona-1,3,5-trien-7-yl)-N,N-dimethyl-4-(4-(4-methylsulfonyl-piperazin-1-yl)-phenyl)-pyrrolidin-3-amine MollogP 4.425 PSA 39.69 No. of chiral centres 3 No. of rotatable bonds 5 No. of hydrogen bond acceptors 7 No. of hydrogen bond donors 0 Physical and chemical properties for A-395N Molecular weight 442.2 Molecular formula C24H34N4O2S IUPAC name 1-benzyl-N,N-dimethyl-4-(4-(4-methylsulfonyl-piperazin-1-yl)-phenyl)-pyrrolidin-3-amine MollogP 3.158 PSA 40.58 No. of chiral centres 2 No. of rotatable bonds 6 No. of hydrogen bond acceptors 7 No. of hydrogen bond donors 0 SMILES: A-395: CN(C)[C@H]1CN(C[C@@H]1c1ccc(cc1)N1CCN(CC1)S(C)(=O)=O)C1CCc2cccc(c12)F A-395N: CN(C)[C@@H]1CN(Cc2ccccc2)C[C@H]1c1ccc(cc1)N1CCN(CC1)S(C)(=O)=O InChI: A-395: InChI=1S/C26H35FN4O2S/c1-28(2)25-18-30(24-12-9-20-5-4-6-23(27)26(20)24)17-22(25)19-7-10-21(11-8-19)29-13-15-31(16-14-29)34(3,32)33/h4-8,10-11,22,24-25H,9,12-18H2,1-3H3/t22-,24?,25+/m1/s1 A-395N: InChI=1S/C24H34N4O2S/c1-25(2)24-19-26(17-20-7-5-4-6-8-20)18-23(24)21-9-11-22(12-10-21)27-13-15-28(16-14-27)31(3,29)30/h4-12,23-24H,13-19H2,1-3H3/t23-,24+/m0/s1 InChIKey: A-395: REVJNSVNICWODC-KIDMSAQOSA-N A-395N: VEABDKBNQNHHCB-BJKOFHAPSA-N Selectivity ProfileSelectivity Selectivity of A-395 against 32 methyltransferase enzymes Selectivity of a) A-395 and b) A-395N relative to epigenetic readers using a thermal shift assay A-395 inhibits the activity of the PRC2 complex in cells – high-content screening assay of RD Rhabdoid tumor cell line treated for 72 hours Co-crystal structures Please wait whilst the interactive viewer is loaded! Main features 1. Binding of A-395 to EED 2. Close-up shot of A-395 showing H-bonds. 3. A-395 superimposed with structure of PRC2 complex with Jarid2 peptide (EED site) and H3K27M in EZH2 catalytic site. EED in blue, EZH2 in gray, and SUZ12 in yellow. 4. Close-up of overlay of A-395 with Jarid2 peptide in PRC2 complex. References Yupeng He, Sujatha Selvaraju, Michael L Curtin, et al. The EED protein–protein interaction inhibitor A-395 inactivates the PRC2 complex . Nature Chem. Biol.(2017)
