About A public-private partnership that supports the discovery of new medicines through open access research. About SGC Partners Research Laboratories SGC-Toronto SGC-UNC SGC-Frankfurt SGC-Karolinska SGC-Neuro Governance Collaborators Open Science Equity, Diversity and Inclusion Impact Science and Resources Proteins Structure Gallery Expression Vectors Plasmids Constructs Production Protocols ChromoHub Phylogenetic Trees UbiHub Phylogenetic Trees ChemBioPort Chemical Probes Program CHEMICAL PROBES Donated chemical probes Chemical Handles Human Kinase Chemical Probe Program ANTIBODIES TARGET ENABLING PACKAGES (TEPs) TISSUE PLATFORM Open Lab Notebooks Open Chemistry Network PUBLICATIONS People Global Aled Edwards, CEO Susan McCormick, CFO Max Morgan, General Counsel and Public Policy Director SGC Toronto Cheryl Arrowsmith Dalia Barsyte-Lovejoy Takis Prinos Matthieu Schapira Levon Halabelian Rachel Harding SGC UNC Tim Willson Alison Axtman David Drewry Peter J. Brown Rafael M. Couñago SGC UCL Mat Todd SGC Karolinska Michael Sundström Opher Gileadi SGC Frankfurt Stefan Knapp Susanne Müller-Knapp Thomas Hanke Vladimir Rogov Krishnal Saxena Andreas Joerger Václav Němec Andreas Krämer Sandra Röhm SGC Neuro Edward Fon Thomas Durcan Jean-François Trempe Ziv Gan-Or Peter McPherson Carl Laflamme Roxanne Lariviere News & Outreach News & Events News from SGC Press Releases Events Blog Tweets by thesgconline Careers
A public-private partnership that supports the discovery of new medicines through open access research.
UNC0642 A potent, selective inhibitor of G9a/GLP with improved PK propertiesThis probe is available from Cayman Chemical, Tocris and Sigma For any inquiries please contact proberequests@thesgc.org.group newOverview G9a (EHMT2) and GLP (EHMT1) catalyze the mono and dimethylation of lysine 9 of histone 3 (H3K9) and other non-histone substrates such as p53 and WIZ. Here we present UNC0642, a G9a/GLP chemical probe with improved PK properties relative to UNC0638. UNC0642 exhibits an in vitro IC50 <15 nM with selectivity > 100-fold over 13 other HMTs and selected representatives of kinases, ion channels, 7TMs, and other epigenetic proteins. In cells, UNC0642 results in a potent reduction of H3K9me2 in MDA MB231 cells with IC50 = 106 nM. Properties 2-(4,4-difluoropiperidin-1-yl)-6-methoxy-N-[1-(propan-2-yl)piperidin-4-yl]-7-[3-(pyrrolidin-1-yl)propoxy]quinazolin-4-amine Click here to download SDF file Physical and chemical properties Molecular weight 546.3 Molecular formula C29H44F2N6O2 IUPAC name 2-(4,4-difluoropiperidin-1-yl)-6-methoxy-N-[1-(propan-2-yl)piperidin-4-yl]-7-[3-(pyrrolidin-1-yl)propoxy]quinazolin-4-amine clogP 5.96 PSA 54.6 A SMILES: N1C2=C(C=C(C(=C2)OCCCN(CCC2)C2)OC)C(=NC=1N(CCC(C1)(F)F)C1)NC(CCN(C(C)C)C1)C1 InChI: InChI=1S/C29H44F2N6O2/c1-21(2)36-14-7-22(8-15-36)32-27-23-19-25(38-3)26(39-18-6-13-35-11-4-5-12-35)20-24(23)33-28(34-27)37-16-9-9(30,31)10-17-37/h19-22H,4-18H2,1-3H3,(H,32,33,34) InChIKey: RNAMYOYQYRYFQY-UHFFFAOYSA-N Selectivity Profile Below we show UNC0642 is equipotent to UNC0638 in the G9a in vitro assay: G9a: Ki = 4 ± 2 (nM) Competitive with peptide substrate Non-competitive with SAM cofactor Similar potency as UNC0638 (Ki = 3 nM) In addition, UNC0642 has been shown to be inactive versus 50 kinases at 10 uM and has a similar GPCR selectivity as UNC0638. Cell-based Assay Data a i.p. administration of a single 5 mg/kg dose in male Swiss Albino mice. 8 time points and 3 animals per time point Materials and Methods MDA-MB231 cells were cultured in RPMI with 10% FBS and MCF7 cells cultured in DMEM with 10% FBS. MTT Toxicity Assay Cells were grown in the presence or absence of inhibitors for stated amount of time. The media was removed and replaced with DMEM 10% FBS without phenol red supplemented with 1mg/ml of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and incubated for 1-2h. Live cells reduce yellow MTT to purple formazan. The resulting formazan was solubilized in acidified isopropanol and 1% Triton and absorbance measured at 570nm, corrected for 650nm background. In-Cell Western (ICW) Cells were grown in 96-well plates in the presence of inhibitors as stated in figures. Media was removed by flicking and 2% formaldehyde in PBS added for 15min. After five washes with 0.1% Triton X100 in PBS, cells were blocked for 1h with 1% BSA in PBS. Three out of four replicates were exposed to primary H3K9m2 antibody, Abcam #1220 at 1/800 dilution in 1% BSA, PBS for 2h. One replicate was reserved for background control. The wells were washed five times with 0.1% Tween 20 in PBS, then secondary IR800 conjugated antibody (LiCor) and DNA-intercalating dye, DRAQ5 (LiCor) added for 1h. After 5 washes with 0.1% Tween 20 in PBS, the plates were read on Odyssey (LiCor) scanner at 800nm (H3K9m2 signal; 764nm excitation) and 700nm (DRAQ5 signal; 683nm excitation). Fluorescence intensity was quantified, normalized to background and DRAQ5 signal expressed as percentage of control. ReferencesDiscovery of an in Vivo Chemical Probe of the Lysine Methyltransferases G9a and GLP; Feng Liu, Dalia Barsyte-Lovejoy, Fengling Li, Yan Xiong, Victoria Korboukh, Xi-Ping Huang, Abdellah Allali-Hassani, William P. Janzen, Bryan L. Roth, Stephen V. Frye, Cheryl H. Arrowsmith, Peter J. Brown, Masoud Vedadi, and Jian Jin; J. Med. Chem., 2013, 56 (21), pp 8931–8942.