About A public-private partnership that supports the discovery of new medicines through open access research. Mission and Philosophy Partners Governance Collaborators Laboratories Key Achievements Research Collaborations and Communications FAQs Science TARGET ENABLING PACKAGES (TEPs) TISSUE PLATFORM PROTEIN STRUCTURES PUBLICATIONS CHEMICAL PROBES Donated chemical probes Biological probes Epigenetics Probes Collection Human Kinase Chemical Probe Program Probes against other protein classes RECOMBINANT ANTIBODIES Structure-guided Drug Discovery Coalition (SDDC) SDDC Publications CREATE ChemNET Research Program Trainees Open Access Learning Impact Faculty & Industry Participants Structural & Chemical Biology Bootcamp Events Structural Biology Programme Structural Parasitology Rare Diseases Genome Integrity and Neurodegeneration Growth Factor Signalling Integral Membrane Proteins Metabolic Enzymes Phosphorylation-Dependent Signalling Protein Kinases Ubiquitin Biology Chemical Biology Antibodies Protein Families Protein Methyltransferases Histone Deacetylases Histone Acetyltransferases Demethylases Bromodomains Methyl Lysine Readers Probe Development Screening Platforms Epigenetic Cell Assays Fragment Based Screening Informatics Medicinal Chemistry ChromoHub Phylogenetic Trees UbiHub Phylogenetic Trees Histone Tails Epigenetics Pocketome Technological Science Biotechnology Biophysics Screening Platforms Research Informatics Protein Crystallography Medicinal Chemistry Reagents & Resources Reagents Chemical Probes Antibodies SGC Vectors SGC Plasmids SGC Constructs Resources Structure Gallery Protein Production Protocols ChromoHub Phylogenetic Trees UbiHub Phylogenetic Trees Histone Tails Fragment Screening Crystal Forms Fragalysis MichaelaNGLo Technologies High Throughput Protein Crystallisation Lex Bubbling System Frapid People Global SGC Director Aled Edwards Susan McCormick Max Morgan Academic Collaborators Industrial/ SME collaborators Non-Scientists Toronto Cheryl Arrowsmith Dalia Barsyte-Lovejoy Peter J. Brown Jinrong Min Takis Prinos Matthieu Schapira Masoud Vedadi Levon Halabelian Biotechnology Crystallography UNC Tim Willson Bill Zuercher Carrow Wells Alison Axtman David Drewry Karolinska Michael Sundström Susanne Gräslund Louise Berg Per-Johan Jakobsson Frankfurt Stefan Knapp Susanne Müller-Knapp News & Outreach News & Events SGC News Symposia & Workshops Blog Press Releases Public Events Public Engagement SGC Languages General Public Open Lab Notebooks Schools Media Patient Groups Governments TOronto Ubiquitin Club And Network (TOUCAN) Careers Tweets by thesgconline
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)
