06.05.2013

Boehringer-Ingelheim joins SGC

by: SGC

INGELHEIM, Germany (6th May, 2013) – Today Boehringer Ingelheim announced that it has joined the Structural Genomics Consortium (SGC). The consortium intends to promote research into protein structures and epigenetics that could pave the way for the development of novel therapies for previously uncurable diseases.As a member of the SGC, Boehringer Ingelheim will help fund precompetitive drug research aimed at bringing new, more effective medicines to patients faster. The SGC is based at the Universities of Toronto, Canada, and Oxford, England.

(R)-PFI-2 A probe for SETD7

This probe hydrochloride) is available from Sigma (including its negative control), Tocris (including its negative control) and Cayman Chemical (including its negative control).

  • overview
  • properties
  • selectivity profile
  • cell based assay data
  • references
  • co-crystal structures
overview
Probe Negative control

 

(R)-PFI-2

 

(S)-PFI-2

A collaboration between the SGC and Pfizer has resulted in the discovery of (R)-PFI-2, a chemical Probe for SETD7.  PFI-2 is a potent inhibitor of SETD7 with IC50 2 nM and 1000-fold selectivity over other methyltransferases and other non-epigenetic targets.  PFI-2 has been shown to bind to SETD7 by ITC (Kd=18nM) and biotinylated PFI-2 interacts with SETD7 in pull-down studies.  Its enantiomer (S)-PFI-2 is 500-fold less active making it an excellent negative control.  Treatment of low-density MEFs with (R)-PFI-2 resulted in higher nuclear YAP levels indicating an effect on the Hippo pathway.

properties
Probe Negative control

 

(R)-PFI-2

 

(S)-PFI-2

Physical and chemical properties for (S)-PFI-2
Molecular weight499.2
Molecular formulaC23H25F4N3O3S
MollogP3.504
PSA70.67
No. of chiral centres1
No. of rotatable bonds8
No. of hydrogen bond acceptors8
No. of hydrogen bond donors2
Physical and chemical properties for (R)-PFI-2
Molecular weight499.2
Molecular formulaC23H25F4N3O3S
MollogP3.504
PSA70.67
No. of chiral centres1
No. of rotatable bonds8
No. of hydrogen bond acceptors8
No. of hydrogen bond donors2
  • SMILES:
  • (S)-PFI-2: FC(F)(F)C1=CC(C[C@H](NS(C2=CC(F)=C3CNCCC3=C2)(=O)=O)C(N4CCCC4)=O)=CC=C1
  • (R)-PFI-2: FC(F)(F)C1=CC(C[C@@H](NS(C2=CC3=C(C(F)=C2)CNCC3)(=O)=O)C(N4CCCC4)=O)=CC=C1
  • InChI:
  • (S)-PFI-2: InChI=1S/C23H25F4N3O3S/c24-20-13-18(12-16-6-7-28-14-19(16)20)34(32,33)29-21(22(31)30-8-1-2-9-30)11-15-4-3-5-17(10-15)23(25,26)27/h3-5,10,12-13,21,28-29H,1-2,6-9,11,14H2/t21-/m0/s1
  • (R)-PFI-2: InChI=1S/C23H25F4N3O3S/c24-20-13-18(12-16-6-7-28-14-19(16)20)34(32,33)29-21(22(31)30-8-1-2-9-30)11-15-4-3-5-17(10-15)23(25,26)27/h3-5,10,12-13,21,28-29H,1-2,6-9,11,14H2/t21-/m1/s1
  • InChIKey:
  • (S)-PFI-2: JCKGSPAAPQRPBW-NRFANRHFSA-N
  • (R)-PFI-2: JCKGSPAAPQRPBW-OAQYLSRUSA-N
selectivity profile

in vitro potency
cell based assay data
references

Barsyte-Lovejoy D, Li F, Oudhoff MJ, Tatlock JH, Dong A, Zeng H, Wu H, Freeman SA, Schapira M, Senisterra GA, Kuznetsova E, Marcellus R, Allali-Hassani A, Kennedy S, Lambert JP, Couzens AL, Aman A, Gingras AC, Al-Awar R, Fish PV, Gerstenberger BS, Roberts L, Benn CL, Grimley RL, Braam MJ, Rossi FM, Sudol M, Brown PJ, Bunnage ME, Owen DR, Zaph C, Vedadi M, Arrowsmith CH. (R)-PFI-2 is a potent and selective inhibitor of SETD7 methyltransferase activity in cells. Proc Natl Acad Sci U S A. 2014 Sep 2;111(35):12853-8. doi: 10.1073/pnas.1407358111. Epub 2014 Aug 18.

pk properties
co-crystal structures

Please wait whilst the interactive viewer is loaded!

Main features

  • PFI-2 efficiently exploits the substrate binding site of SETD7.
  • PFI-2 interacts directly with the departing methyl group of SAM.
  • PFI-2 occupies the lysine binding site of SETD7.
synthetic schemes
materials and methods

UNC1999 A chemical probe for EZH2/1

The probe UNC1999 is available from Cayman Chemical, Sigma and Tocris.

  • overview
  • properties
  • selectivity profile
  • cell based assay data
  • references
overview
Probe Negative control

 

UNC1999

 

UNC2400

Biotinylated UNC1999

UNC2399

The histone H3-lysine 27 (H3K27) methyltransferase EZH2 plays a critical role in regulating gene expression, and its aberrant activity is linked to the onset and progression of cancer. A collaboration between the SGC and the Center for Integrative Chemical Biology and Drug Discovery (CICBDD) at the University of North Carolina at Chapel Hill has resulted in the discovery of UNC1999. UNC1999 inhibits EZH2 with an IC50 of 2nM and is over 1000-fold selective for other HMTs except EZH1 (22-fold selectivity). UNC1999 inhibits H3K27 methylation in MCF10A cells with an IC50 of 124nM as measured by immunofluorescence. A dimethylated version, UNC2400, was also used as a negative control compound in key experiments.

properties
UNC1999

N-[(6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl]-1-(propan-2-yl)-6-{6-[4-(propan-2-yl)piperazin-1-yl]pyridin-3-yl}-1H-indazole-4-carboxamide

Click here to download the SD file

Physical and chemical properties
Molecular weight569.74
Molecular formulaC33H43N7O2
IUPAC nameN-[(6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl]-1-(propan-2-yl)-6-{6-[4-(propan-2-yl)piperazin-1-yl]pyridin-3-yl}-1H-indazole-4-carboxamide
logP4.01
PSA95.39
  • SMILES
  • CCCC(C=C(N1)C)=C(CNC(C2=CC(C3=CC=C(N4CCN(C(C)C)CC4)N=C3)=CC5=C2C=NN5C(C)C)=O)C1=O
  • InChi
  • InChI=1S/C33H43N7O2/c1-7-8-24-15-23(6)37-33(42)28(24)19-35-32(41)27-16-26(17-30-29(27)20-36-40(30)22(4)5)25-9-10-31(34-18-25)39-13-11-38(12-14-39)21(2)3/h9-10,15-18,20-22H,7-8,11-14,19H2,1-6H3,(H,35,41)(H,37,42)
  • InChiKey
  • InChIKey=DPJNKUOXBZSZAI-UHFFFAOYSA-N
selectivity profile

UNC1999 is selective for EZH2 over 15 other methyltransferases and proteins in other target classes

  • Less than 20% inhibition on 50 kinase targets @ 10 µM.
  • Less than 50% inhibition on 40 7TM targets @ 10 µM.
  • Greater than 50% inhibition of 4 7TM targets @ 10 µM.
    • H3 (Ki 300 nM), NET (Ki 1500nM), Sigma 1(Ki 4700 nM), Sigma 2 (Ki 65 nM)
    • No functional activity on H3, functional assay not available for Sigma 2
in vitro potency
cell based assay data

UNC1999 is Cellularly Active

A. Treatment of MCF10A cells for 3 days with UNC1999 shows a dose-related decrease in H3K27me3 which is unrelated to cell viability. 
B. UNC2400 did not show any dose-related decrease in H3K27me3.

UNC1999 Kills DB cells with Y641N mutation

DB cells harbor the EZH2 Y641N mutation

C. 8-day treatment of UNC1999 but not UNC2400 kills DB cells.
D. Western blotting of EZH2, total H3 and H3K27me3 following 3-day treatment of UNC1999 shows no change in H3 or EZH2, but significant change in H3K27me3

references

An Orally Bioavailable Chemical Probe of the Lysine Methyltransferases EZH2 and EZH1
 

Kyle D. Konze, Anqi Ma, Fengling Li, Dalia Barsyte-Lovejoy, Trevor Parton, Christopher J.MacNevin, Feng Liu, Cen Gao, Xi-Ping Huang, Ekaterina Kuznetsova, Marie Rougie, Alice Jiang, Samantha G. Pattenden, Jacqueline L. Norris, Lindsey I. James, Bryan L Roth, Peter J. Brown, Stephen V. Frye, Cheryl H. Arrowsmith, Klaus M. Hahn, Gang Greg Wang, Masoud Vedadi, and Jian Jin.

ACS Chem. Biol., 2013, 8 (6), pp 1324–1334 

DOI: 10.1021/cb400133j • Publication Date (Web): 08 Apr 2013

pk properties
co-crystal structures
synthetic schemes
materials and methods
02.04.2013

Novel structure of a human membrane enzyme sheds light on molecular mechanisms of rare ageing disorders and metabolic syndromes

by: SGC

Laminopathies are a group of rare genetic disorders caused by mutations in genes encoding proteins involved in the building and maintenance of the nuclei of cells. Severe laminopathies such as the premature ageing syndrome pregeria leads to death in the teens from heart disease and other symtoms normally found in people in their 80s. The structure of the nucleus is maintained by a class of proteins called lamins. When one of these proteins, prelamin A is not properly processed by an enzyme ZMPSTE24, laminopathies occur.

I-CBP112 A CREBBP/EP300-selective chemical probe

This probe is available from Sigma and Cayman Chemical.

This probe (as hydrochloride) is available from Cayman Chemical.

  • overview
  • properties
  • selectivity profile
  • cell based assay data
  • references
  • co-crystal structures
overview
Probe

I-CBP112

CREBBP (CBP) and EP300 are general transcriptional co-activators, which are involved in many biological processes like maintenance of genomic stability by affecting DNA replication and DNA repair as well as cell growth, transformation and development. They  also plays and essential role in neuronal plasticity/ memory formation, hematopoiesis and energy homeostasis as demonstrated in a variety of mouse models. By acetylation of non-histone proteins CREBBP can have a positive or negative effect on transcriptional regulation affecting protein- protein interactions, protein-DNA interactions, nuclear retention or protein half-life.

Mutations of CREBBP and EP300 lead to Rubinstein-Taybi syndrome (RTS), characterised by growth retardation, facial abnormalities, organ abnormalities, mental retardation and proneness to tumors. Chromosomal translocations of CREBBP or EP300 with MOZ, MLL have been observed in acute myeloid leukemia. CREBBP has also been associated with Amyotrophic lateral sclerosis (ALS), Lou Gherig’s disease, a neurodegenerative disease with progressive degeneration of motor neurons in the brain and spinal cord, Alzheimer's disease and polyglutamine diseases such as Spinal and Bulbar Muscular Atrophy and Huntington’s disease.

We have developed an inhibitor, I-CBP112, against the CREBBP and EP300 Bromodomains.

In vitro Potency
AssayIC50/Kd
CREBBP (Alphascreen)0.170
CREBBP (BLI)0.170
CREBBP (ITC)0.151
EP300 (ITC)0.625
properties
I-CBP112
1-[7-(3,4-dimethoxyphenyl)-9-{[(3S)-1-methylpiperidin-3-yl]methoxy}-2,3,4,5-tetrahydro-1,4-benzoxazepin-4-yl]propan-1-one
Click here to download SDF file
Physical and chemical properties
Molecular weight468.3
clogP4.5
PSA50.2
Storage2-8°C as powder. NB making aliquots rather than freeze-thawing is recommended
DissolutionSoluble in DMSO at least up to 50 mM
  • SMILES:
  • CN1CCC[C@H](COC2=C3C(CN(CCO3)C(CC)=O)=CC(C4=CC=C(C(OC)=C4)OC)=C2)C1
  • InChI:
  • InChI=1S/C27H36N2O5/c1-5-26(30)29-11-12-33-27-22(17-29)13-21(20-8-9-23(31-3)24(14-20)32-4)15-25(27)34-18-19-7-6-10-28(2)16-19/h8-9,13-15,19H,5-7,10-12,16-18H2,1-4H3/t19-/m0/s1
  • InChIKey:
  • YKNAKDFZAWQEEO-IBGZPJMESA-N
selectivity profile

Selectivity against other Bromodomains

BLI@ 0.2 and 1 µM:

No interaction was detected by BLI for the selectivity panel comprising bromodomains:

ATAD2

BAZ2B

BRD2(2)

BRD4 (1)

PB1(5)

PCAF

PHIP(2)

TIF1α

Against the entire bromodomain family, weak cross-reactivity was only observed for the BET bromodomains. 

in vitro potency
cell based assay data
Probe

I-CBP112

CREBBP (3x bromodomain) (FRAP assay) - Accelerated FRAP recovery at 1 µM.

No significant cytotoxicity up to 50 μM in U2OS cells.

references

Generation of a selective small molecule inhibitor of the CBP/p300 bromodomain for leukemia therapy.
Picaud S, Fedorov O, Thanasopoulou A, Leonards K, Jones K, Meier J, Olzscha H, Monteiro O, Martin S, Philpott M, Tumber A, Filippakopoulos P, Yapp C, Wells C, Hing Che K, Bannister A, Robson S, Kumar U, Parr N, Lee K, Lugo D, Jeffrey P, Taylor S, Vecellio ML, Bountra C, Brennan P, O'Mahony A, Velichko S, Muller S, Hay D, Daniels DL, Urh M, La Thangue NB, Kouzarides T, Prinjha R, Schwaller J, Knapp S.
Cancer Res. 2015;75(23):5106-19.

pk properties
co-crystal structures

I-CBP112 has been co-crystallised with CREBBP and is deposited in the PDB - 4NR6

synthetic schemes
materials and methods

SGC-CBP30 A CREBBP/EP300-selective chemical probe

This probe is available from Tocris, Sigma and Cayman Chemical

  • overview
  • properties
  • selectivity profile
  • cell based assay data
  • references
  • co-crystal structures
overview
Probe Negative control

 

SGC-CBP30

 

BDOIA513 

In vitro Potency
AssayIC50/Kd µM
CREBBP (Alphascreen)0.069
CREBBP (BLI)0.041
CREBBP (ITC)0.021
EP300 (ITC)0.038

CREBBP (CBP) and EP300 are general transcriptional co-activators, which are involved in many biological processes like maintenance of genomic stability by affecting DNA replication and DNA repair as well as cell growth, transformation and development. They also play and essential role in neuronal plasticity/ memory formation hematopoiesis and energy homeostasis as demonstrated in a variety of mouse models. They possess both acetyl-transferase enzymatic and bromodomain containing regions. Through acetylation of non-histone proteins CREBBP can have a positive or negative effect on transcriptional regulation by affecting protein- protein interactions, protein-DNA interactions, nuclear retention or protein half-life.

Mutations of CREBBP and EP300 lead to Rubinstein-Taybi syndrome (RTS), characterised by growth retardation, facial abnormalities, organ abnormalities, mental retardation and altered tumor susceptibility. Chromosomal translocations of CREBBP or EP300 with MOZ, MLL have been observed in acute myeloid leukemia.

CREBBP has also been associated with Amyotrophic Lateral Sclerosis (ALS) or Lou Gehrig’s disease, a neurodegenerative disease with progressive degeneration of motor neurons in the brain and spinal cord, Alzheimer's disease and poly glutamine repeat diseases such as Spinal and Bulbar Muscular Atrophy and Huntington’s disease.

We have developed an inhibitor, SGC-CBP30* against the CREBBP and EP300 Bromodomains.

* This is officially pronounced SGC-CBP-three-oh

SGC-CBP30N (BDOIA513) is the negative control for SGC-CBP30. This compound has moderate activity for CREBBP (Thermal shift 2°C compared to 9.7°C for SGC-CBP30). This compound is inactive against BRD4 (Thermal shift 0.4°C compared to 1.8°C for SGC-CBP30)

properties
SGC-CBP30
8-(3-chloro-4-methoxy-phenethyl)-4-(3,5-dimethyl-isoxazol-4-yl)-9-(2-(morpholin-4-yl)-propyl)-7,9-diaza-bicyclo[4.3.0]nona-1(6),2,4,7-tetraene
Click here to download SDF file
Physical and chemical properties
Molecular weight508.2
Molecular formulaC28H33ClN4O3
IUPAC name8-(3-chloro-4-methoxy-phenethyl)-4-(3,5-dimethyl-isoxazol-4-yl)-9-(2-(morpholin-4-yl)-propyl)-7,9-diaza-bicyclo[4.3.0]nona-1(6),2,4,7-tetraene
logP4.89
PSA51.8
Storage2-8°C as powder. NB making aliquots rather than freeze-thawing is recommended
DissolutionSoluble in DMSO at least up to 50mM
  • SMILES:
  • CC1=NOC(C)=C1C2=CC(N=C(N3C[C@@H](N4CCOCC4)C)CCC5=CC(Cl)=C(OC)C=C5)=C3C=C2
  • InChI:
  • InChI=1S/C28H33ClN4O3/c1-18(32-11-13-35-14-12-32)17-33-25-8-7-22(28-19(2)31-36-20(28)3)16-24(25)30-27(33)10-6-21-5-9-26(34-4)23(29)15-21/h5,7-9,15-16,18H,6,10-14,17H2,1-4H3/t18-/m0/s1
  • InChIKey:
  • InChIKey=GEPYBHCJBORHCE-SFHVURJKSA-N

Measured Properties


(click for a larger version)

selectivity profile

Differential Scanning Fluorimetry (DSF)

Thermal melting experiments were carried out using an Mx3005p Real Time PCR machine (Stratagene). Proteins were buffered in 10 mM HEPES pH 7.5, 500 mM NaCl and assayed in a 96-well plate at a final concentration of 2 µM in 20 µl volume. BDOIA518 was added at a final concentration of 10 µM. SYPRO Orange (Molecular Probes) was added as a fluorescence probe at a dilution of 1:1000. Excitation and emission filters for the SYPRO-Orange dye were set to 465 nm and 590 nm, respectively. The temperature was raised with a step of 3 °C per minute from 25 °C to 96 °C and fluorescence readings were taken at each interval. The temperature dependence of the fluorescence during the protein denaturation process was fit to the Boltzmann equation.


Activity against BRD4(1) by ITC 0.86 mM: 40 fold selective for CBP. (click for larger version)

SGC-CBP30 was profiled against the CEREP panel of Receptors, Ion Channels and Enzymes with the following results

 
(click for larger version)

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(click for larger version)

​

(click for larger version)

in vitro potency
cell based assay data
Activity In cells
CREBBP (3x bromodomain) (FRAP assay)Accelerated FRAP recovery at 1 µM

Moderate cytotoxicity in U2OS cells and HeLa cells.

references

Discovery and Optimization of Small-Molecule Ligands for the CBP/p300 Bromodomains

Duncan A. Hay, Oleg Fedorov, Sarah Martin, Dean C. Singleton, Cynthia Tallant,Christopher Wells, Sarah Picaud, Martin Philpott, Octovia P. Monteiro, Catherine M. Rogers, Stuart J. Conway, Timothy P. C. Rooney, Anthony Tumber, Clarence Yapp,Panagis Filippakopoulos, Mark E. Bunnage,Susanne Müller, Stefan Knapp, Christopher J. Schofield, and Paul E. Brennan

J. Am. Chem. Soc., 2014, doi: 10.1021/

 

CBP30, a selective CBP/p300 bromodomain inhibitor, suppresses human Th17 responses.

Hammitzsch A, Tallant C, Fedorov O, O'Mahony A, Brennan PE, Hay DA, Martinez FO, Al-Mossawi MH, de Wit J, Vecellio M, Wells C, Wordsworth P, Müller S, Knapp S, Bowness P.

Proc Natl Acad Sci U S A. 2015, 112(34), 10768-73. doi: 10.1073/pnas.1501956112

pk properties
co-crystal structures

SGC-CBP30 has been co-crystallised with CREBBP and is deposited in the PDB - 4NR7

synthetic schemes
materials and methods

Bromosporine (BSP)

This compound is available from Tocris, Sigma and Cayman Chemical

  • overview
  • properties
  • selectivity profile
  • cell based assay data
  • references
  • synthetic schemes
overview
Probe

Bromosporine (BSP)

Bromodomains (BRDs) are protein interaction modules that read epigenetic marks recognizing ε-N-lysine acetylation motifs. The conserved BRD fold contains a deep, largely hydrophobic acetyllysine binding site, an attractive pocket for the development of small, pharmaceutically active molecules. BRDs have an important role in the targeting of chromatin-modifying enzymes to specific sites, including methyltransferases, HATs and transcription factors and regulate diverse biological processes from cell proliferation and differentiation to energy homeostasis and neurological processes.


Co-crystal structure of Bromosporine with BRD4(1)

Proteins that contain BRDs have been implicated in the development of a large variety of diseases, including various cancers, inflammatory diseases and neurological diseases and the therapeutic potential of bromodomain inhibition has been shown in several of these diseases, such as HIV, cancer and inflammation. Bromosporine is a broad spectrum inhibitor for bromodomains and as such will be very useful in elucidating further biological roles of reader domains as well as a tool for the validation of functional assays.
properties

ethyl N-[6-(3-methanesulfonamido-4-methylphenyl)-3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-8-yl]carbamate
Click here to download SDF file

Physical and chemical properties
Molecular weight404.44
Molecular formulaC17H20N6O4S
IUPAC Nameethyl N-[6-(3-methanesulfonamido-4-methylphenyl)-3-methyl-[1,2,4]triazolo[4,3-b]pyridazin-8-yl]carbamate
clogP0.78
PSA105.64
Storage2-8°C as powder. NB making aliquots rather than freeze-thawing is recommended
DissolutionSoluble in DMSO at least up to 50mM
SMILES:

CC1=NN=C2N1N=C(C3=CC=C(C)C(NS(=O)(C)=O)=C3)C=C2NC(OCC)=O

InChI:

InChI=1S/C17H20N6O4S/c1-5-27-17(24)18-15-9-14(21-23-11(3)19-20-16(15)23)12-7-6-10(2)13(8-12)22-28(4,25)26/h6-9,22H,5H2,1-4H3,(H,18,24)

InChIKey:

UYBRROMMFMPJAN-UHFFFAOYSA-N

selectivity profile

Differential Scanning Fluorimetry (DSF)

Thermal melting experiments were carried out using an Mx3005p Real Time PCR machine (Stratagene). Proteins were buffered in 10 mM HEPES pH 7.5, 500 mM NaCl and assayed in a 96-well plate at a final concentration of 2 µM in 20 µl volume. Bromosporine was added at a final concentration of 10 µM. SYPRO Orange (Molecular Probes) was added as a fluorescence probe at a dilution of 1:1000. Excitation and emission filters for the SYPRO-Orange dye were set to 465 nm and 590 nm, respectively. The temperature was raised with a step of 3 °C per minute from 25 °C to 96 °C and fluorescence readings were taken at each interval. The temperature dependence of the fluorescence during the protein denaturation process was approximated by the equation (displayed below) where ΔuG(T) is the difference in unfolding free energy between the folded and unfolded state, R is the gas constant and yF and yU are the fluorescence intensity of the probe in the presence of completely folded and unfolded protein respectively. The baselines of the denatured and native states were approximated by a linear fit. The observed temperature shifts, ΔTmobs, were recorded as the difference between the transition midpoints of sample and reference wells containing protein without ligand in the same plate and determined by non-linear least squares fit.

Selectivity
BromodomainThermal melt
10 µM
ΔTm
BRD2(1)4.4
BRD2(2)5.9
BRD3(1)5.3
BRD3(2)5.9
BRD4(1)6.9
BRD4(2)6.2
BRDT(1)7.3
BRDT(2)5.2
CECR28.3
PB1(5)0.4
TAF1(1)1.3
TAF1(2)5.2
TAF1L(1)0.9
TAF1L(2)0.9
BAZ2A1.2
TIF1α0.4
ATAD2-0.2
BRD93.9
CREBBP3.4
in vitro potency
cell based assay data
Potency in Cells
BRD4Accelerated FRAP recovery at 1 µM
CREBBPAccelerated FRAP recovery at 1 µM
TIF1αInactive at 10 µM
BAZ2AInactive at 10 µM
SMARCA2Inactive at 10 µM

Bromosporine shows moderate cytotoxicity in HeLa cells at 18 µM

references

 

Promiscuous targeting of bromodomains by bromosporine identifies BET proteins as master regulators of primary transcription response in leukemia

Sarah Picaud, Katharina Leonards, Jean-Philippe Lambert, Oliver Dovey, Christopher Wells, Oleg Fedorov, Octovia Monteiro, Takao Fujisawa, Chen-Yi Wang, Hannah Lingard, Cynthia Tallant, Nikzad Nikbin, Lucie Guetzoyan, Richard Ingham, Steven V. Ley, Paul Brennan, Susanne Muller, Anastasia Samsonova, Anne-Claude Gingras, Juerg Schwaller, George Vassiliou, Stefan Knapp and Panagis Filippakopoulos. Science Advances  12 Oct 2016: Vol. 2, no. 10, e1600760. DOI: 10.1126/sciadv.1600760

pk properties
co-crystal structures
synthetic schemes

Scheme 1

Scheme 2

materials and methods
16.11.2012

GlaxoSmithKline releases EZH2 inhibitor as part of SGC epigenetics initiative

by: SGC

GlaxoSmithKline has developed a potent, selective, cell-active EZH2 inhibitor, GSK343, and has made this available as a chemical probe as part of the SGC epigenetics initiative.  The histone H3-lysine 27 (H3K27) methyltransferase EZH2 plays a critical role in regulating gene expression, and its aberrant activity is linked to the onset and progression of cancer.  GSK343 inhibits EZH2 with an IC50 of 4nM and is over 1000-fold selective for other HMTs except EZH1 (60-fold selectivity).

19.09.2012

Pfizer and SGC Oxford receive the first Wellcome Trust Pathfinder Award for Rare Diseases

by: SGC

The Pfizer-SGC project aims to tackle a rare, hereditary metabolic disorder called homocystinuria, which leaves patients unable to metabolise the amino acid methionine. A team of researchers at Pfizer Rare Disease unit will work with Dr Wyatt Yue at the Structural Genomics Consortium at the University of Oxford to investigate methods of restoring normal metabolism and preventing the progression of the disease. The team will study the enzyme that is affected by the disease and investigate ways to restore its function.

GSK343 A potent, selective inhibitor of EZH2 with cellular activity

This probe is available from Sigma, Cayman Chemical and Tocris.

  • overview
  • properties
  • selectivity profile
  • cell based assay data
  • references
overview
Probe

GSK343

The histone H3-lysine 27 (H3K27) methyltransferase EZH2 plays a critical role in regulating gene expression, and its aberrant activity is linked to the onset and progression of cancer.  GlaxoSmithKline has developed a potent, selective, cell-active EZH2 inhibitor, GSK343, and has made this available as a chemical probe as part of the SGC epigenetics initiative.  GSK343 inhibits EZH2 with an IC50 of 4nM and is over 1000-fold selective for other HMTs except EZH1 (60-fold selectivity).  GSK343 inhibits H3K27 methylation in HCC1806 cells with an IC50 of <200nM as measured by immunofluorescence.

properties

N-[(6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl]
-6-[2-(4-methylpiperazin-1-yl)pyridin-4-yl]
-1-(propan-2-yl)-1H-indazole-4-carboxamide
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Physical and chemical properties
Molecular weight541.3
Molecular formulaC31H39N7O2
IUPAC nameN-[(6-methyl-2-oxo-4-propyl-1,2-dihydropyridin-3-yl)methyl]
-6-[2-(4-methylpiperazin-1-yl)pyridin-4-yl]
-1-(propan-2-yl)-1H-indazole-4-carboxamide
logP3.23
PSA95.39
  • SMILES:
  • CCCC1=C(CNC(=O)C2=C3C=NN(C(C)C)C3=CC(=C2)C2=CC=NC(=C2)N2CCN(C)CC2)C(=O)NC(C)=C1
  • InChI:
  • InChI=1S/C31H39N7O2/c1-6-7-23-14-21(4)35-31(40)26(23)18-33-30(39)25-15-24(16-28-27(25)19-34-38(28)20(2)3)22-8-9-32-29(17-22)37-12-10-36(5)11-13-37/h8-9,14-17,19-20H,6-7,10-13,18H2,1-5H3,(H,33,39)(H,35,40)
  • InChIKey:
  • ULNXAWLQFZMIHX-UHFFFAOYSA-N
selectivity profile

in vitro potency
cell based assay data

GSK343 inhibits H3K27 methylation in HCC1806 cells with an IC50 of <200nM as measured by immunofluorescence.


Biological characterization of GSK343. (a) H3K27me3 imaging in HCC1806 cells treated with DMSO or GSK343 for 72 hours; (b) Dose-response of GSK343-treated HCC 1806 cells.

references

Identification of Potent, Selective, Cell-Active Inhibitors of the Histone Lysine Methyltransferase EZH2 

Sharad K. Verma*, Xinrong Tian, Louis V. LaFrance, Céline Duquenne, Dominic P.Suarez, Kenneth A. Newlander, Stuart P. Romeril, Joelle L. Burgess, Seth W. Grant, James A. Brackley, Alan Graves, Daryl A. Scherzer, Art Shu, Christine S. Thompson, Heidi Morgan-Ott, Glenn S. Van Aller, Carl A. Machutta, Elsie Diaz, Yong Jiang, Neil W. Johnson, Steven Knight, Ryan G. Kruger, Michael T. McCabe, Dashyant Dhanak, Peter J. Tummino, Caretha L. Creasy and William H. Miller.  ACS Med. Chem. Lett., 2012, 3 (12), pp 1091–1096 DOI: 10.1021/ml3003346.

pk properties
co-crystal structures
synthetic schemes
materials and methods
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