S-adenosylhomocysteine [Ligand Id: 5265] activity data from GtoPdb and ChEMBL

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ChEMBL ligand: CHEMBL418052 (S-Adenosylhomocysteine, S-Adenosyl Homocysteine, S-Adenosyl-Homocysteine)
  • DNA methyltransferase 1/DNA (cytosine-5)-methyltransferase 1 in Human [ChEMBL: CHEMBL1993] [GtoPdb: 2605] [UniProtKB: P26358]
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  • DNA (cytosine-5-)-methyltransferase 3α/DNA (cytosine-5)-methyltransferase 3A in Human [ChEMBL: CHEMBL1992] [GtoPdb: 2750] [UniProtKB: Q9Y6K1]
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  • DNA (cytosine-5)-methyltransferase 3B in Human [ChEMBL: CHEMBL6095] [UniProtKB: Q9UBC3]
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  • Histamine N-methyltransferase in Human [ChEMBL: CHEMBL2190] [UniProtKB: P50135]
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  • coactivator associated arginine methyltransferase 1/Histone-arginine methyltransferase CARM1 in Human [ChEMBL: CHEMBL5406] [GtoPdb: 1255] [UniProtKB: Q86X55]
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  • enhancer of zeste 1 polycomb repressive complex 2 subunit/Histone-lysine N-methyltransferase EZH1 in Human [ChEMBL: CHEMBL2189116] [GtoPdb: 2835] [UniProtKB: Q92800]
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  • enhancer of zeste 2 polycomb repressive complex 2 subunit/Histone-lysine N-methyltransferase EZH2 in Human [ChEMBL: CHEMBL2189110] [GtoPdb: 2654] [UniProtKB: Q15910]
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  • DOT1 like histone lysine methyltransferase/Histone-lysine N-methyltransferase, H3 lysine-79 specific in Human [ChEMBL: CHEMBL1795117] [GtoPdb: 2650] [UniProtKB: Q8TEK3]
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  • euchromatic histone lysine methyltransferase 2/Histone-lysine N-methyltransferase, H3 lysine-9 specific 3 in Human [ChEMBL: CHEMBL6032] [GtoPdb: 2652] [UniProtKB: Q96KQ7]
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  • euchromatic histone lysine methyltransferase 1/Histone-lysine N-methyltransferase, H3 lysine-9 specific 5 in Human [ChEMBL: CHEMBL6031] [GtoPdb: 2651] [UniProtKB: Q9H9B1]
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  • lysine methyltransferase 2A/Histone-lysine N-methyltransferase MLL in Human [ChEMBL: CHEMBL1293299] [GtoPdb: 2688] [UniProtKB: Q03164]
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  • nuclear receptor binding SET domain protein 2/Histone-lysine N-methyltransferase NSD2 in Human [ChEMBL: CHEMBL3108645] [GtoPdb: 3220] [UniProtKB: O96028]
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  • SET domain containing 7, histone lysine methyltransferase/Histone-lysine N-methyltransferase SETD7 in Human [ChEMBL: CHEMBL5523] [GtoPdb: 2703] [UniProtKB: Q8WTS6]
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  • SET domain bifurcated histone lysine methyltransferase 1/Histone-lysine N-methyltransferase SETDB1 in Human [ChEMBL: CHEMBL2321646] [GtoPdb: 2705] [UniProtKB: Q15047]
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  • SUV39H1 histone lysine methyltransferase/Histone-lysine N-methyltransferase SUV39H1 in Human [ChEMBL: CHEMBL1795118] [GtoPdb: 2715] [UniProtKB: O43463]
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  • SUV39H2 histone lysine methyltransferase/Histone-lysine N-methyltransferase SUV39H2 in Human [ChEMBL: CHEMBL1795177] [GtoPdb: 2716] [UniProtKB: Q9H5I1]
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  • lysine methyltransferase 5C/Histone-lysine N-methyltransferase SUV420H2 in Human [ChEMBL: CHEMBL2321644] [GtoPdb: 2718] [UniProtKB: Q86Y97]
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  • Indolethylamine N-methyltransferase in Human [ChEMBL: CHEMBL2131] [UniProtKB: O95050]
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  • methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit/N6-adenosine-methyltransferase catalytic subunit in Human [ChEMBL: CHEMBL4739695] [GtoPdb: 3181] [UniProtKB: Q86U44]
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  • nicotinamide N-methyltransferase/Nicotinamide N-methyltransferase in Human [ChEMBL: CHEMBL2346486] [GtoPdb: 3205] [UniProtKB: P40261]
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  • SET and MYND domain containing 2/N-lysine methyltransferase SMYD2 in Human [ChEMBL: CHEMBL2169716] [GtoPdb: 2714] [UniProtKB: Q9NRG4]
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  • Phenylethanolamine N-methyltransferase in Human [ChEMBL: CHEMBL4617] [GtoPdb: 2496] [UniProtKB: P11086]
  • Phenylethanolamine N-methyltransferase in Bovine [ChEMBL: CHEMBL2331] [UniProtKB: P10938]
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  • protein arginine methyltransferase 1 /Protein-arginine N-methyltransferase 1 in Human [ChEMBL: CHEMBL5524] [GtoPdb: 1252] [UniProtKB: Q99873]
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  • protein arginine methyltransferase 3/Protein arginine N-methyltransferase 3 in Human [ChEMBL: CHEMBL5891] [GtoPdb: 1254] [UniProtKB: O60678]
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  • protein arginine methyltransferase 5 /Protein arginine N-methyltransferase 5 in Human [ChEMBL: CHEMBL1795116] [GtoPdb: 1256] [UniProtKB: O14744]
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  • protein arginine methyltransferase 7 /Protein arginine N-methyltransferase 7 in Human [ChEMBL: CHEMBL3562175] [GtoPdb: 1258] [UniProtKB: Q9NVM4]
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  • rRNA adenine N-6-methyltransferase in Streptococcus pneumoniae [ChEMBL: CHEMBL2757] [UniProtKB: P21236]
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  • tRNA (guanine-N(1)-)-methyltransferase in Mycobacterium tuberculosis [ChEMBL: CHEMBL4523933] [UniProtKB: P9WFY7]
  • tRNA (guanine-N(1)-)-methyltransferase in Staphylococcus aureus (strain NCTC 8325 / PS 47) [ChEMBL: CHEMBL4523934] [UniProtKB: Q2FZ43]
  • tRNA (guanine-N(1)-)-methyltransferase in Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM14847 / LMG 12228 / 1C / PRS 101 / PAO1) [ChEMBL: CHEMBL4523932] [UniProtKB: Q9HXQ1]
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DB Assay description Assay Type Standard value Standard parameter Original value Original units Original parameter Reference
DNA methyltransferase 1/DNA (cytosine-5)-methyltransferase 1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1993] [GtoPdb: 2605] [UniProtKB: P26358]
ChEMBL Inhibition of human recombinant DNMT1 expressed in Sf9 cells assessed as incorporation of [3H]S-adenosyl methionine into hemimethylated oligonucleotide substrate after 3 hrs by scintillation counting B 5.4 pIC50 4000 nM IC50 J Med Chem (2011) 54: 7663-7677 [PMID:21958292]
ChEMBL Inhibition of human recombinant DNMT1 expressed in baculovirus-insect cell system by scintillation counting B 5.4 pIC50 4000 nM IC50 Bioorg Med Chem (2010) 18: 822-829 [PMID:20006515]
ChEMBL Inhibition of human recombinant DNMT1 expressed in baculovirus infected high five insect cells B 5.7 pIC50 2000 nM IC50 Bioorg Med Chem Lett (2009) 19: 2747-2751 [PMID:19362833]
ChEMBL Inhibition of human Dnmt1 using oligonucleotide 2 as substrate after 5000 sec by micro plate reader based real-time break-light assay B 6.03 pIC50 941 nM IC50 Bioorg Med Chem Lett (2012) 22: 3079-3082 [PMID:22483584]
ChEMBL Inhibition of full length N-terminal His6-tagged human DNMT1 after 1 hr by fluorescence analysis B 6.05 pIC50 900 nM IC50 J Med Chem (2012) 55: 1731-1750 [PMID:22280363]
ChEMBL Inhibition of human recombinant DNMT1 B 6.1 pIC50 800 nM IC50 Bioorg Med Chem Lett (2009) 19: 2742-2746 [PMID:19364644]
ChEMBL Inhibition of DNMT1 (unknown origin) using biotinylated substrate using [3H]-SAM after 1 hr by scintillation proximity assay B 6.22 pIC50 600 nM IC50 Bioorg Med Chem Lett (2019) 29: 826-831 [PMID:30704813]
ChEMBL Inhibition of DNMT1 (unknown origin) B 6.7 pIC50 200 nM IC50 Bioorg Med Chem Lett (2016) 26: 4518-4522 [PMID:27485386]
ChEMBL Inhibition of recombinant human DNMT1 using poly(dl-dC) as substrate by hotspot assay B 7.15 pIC50 71 nM IC50 Eur J Med Chem (2020) 189: 112023-112023 [PMID:31978781]
DNA (cytosine-5-)-methyltransferase 3α/DNA (cytosine-5)-methyltransferase 3A in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1992] [GtoPdb: 2750] [UniProtKB: Q9Y6K1]
ChEMBL Inhibition of human full length DNMT3A expressed in Sf9 cells B 4.3 pIC50 >50000 nM IC50 ACS Med Chem Lett (2015) 6: 408-412 [PMID:25893041]
DNA (cytosine-5)-methyltransferase 3B in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL6095] [UniProtKB: Q9UBC3]
ChEMBL Inhibition of human full length DNMT3B expressed in Sf9 cells B 4.3 pIC50 >50000 nM IC50 ACS Med Chem Lett (2015) 6: 408-412 [PMID:25893041]
ChEMBL Inhibition of human recombinant DNMT3b2 expressed in baculovirus infected high five insect cells B 6.52 pIC50 300 nM IC50 Bioorg Med Chem Lett (2009) 19: 2747-2751 [PMID:19362833]
ChEMBL Inhibition of human recombinant DNMT3B expressed in baculovirus-insect cell system by scintillation counting B 6.6 pIC50 250 nM IC50 Bioorg Med Chem (2010) 18: 822-829 [PMID:20006515]
ChEMBL Inhibition of human recombinant DNMT3b2 B 6.7 pIC50 200 nM IC50 Bioorg Med Chem Lett (2009) 19: 2742-2746 [PMID:19364644]
Histamine N-methyltransferase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2190] [UniProtKB: P50135]
ChEMBL Inhibition constant was evaluated against Histamine N-methyl-transferase B 4.74 pKi 18100 nM Ki J Med Chem (1985) 28: 478-482 [PMID:3981540]
ChEMBL Inhibition constant was evaluated against Histamine N-methyl-transferase B 4.98 pKi 10500 nM Ki J Med Chem (1985) 28: 478-482 [PMID:3981540]
coactivator associated arginine methyltransferase 1/Histone-arginine methyltransferase CARM1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5406] [GtoPdb: 1255] [UniProtKB: Q86X55]
ChEMBL Inhibition of CARM1 B 6.07 pKi 860 nM Ki J Med Chem (2012) 55: 8066-8074 [PMID:22924785]
ChEMBL Inhibition of CARM1 (unknown origin) B 6.4 pKi 400 nM Ki J Med Chem (2013) 56: 8972-8983 [PMID:23879463]
enhancer of zeste 1 polycomb repressive complex 2 subunit/Histone-lysine N-methyltransferase EZH1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2189116] [GtoPdb: 2835] [UniProtKB: Q92800]
ChEMBL Inhibition of EZH1 (unknown origin) by HMT assay B 5.19 pIC50 6400 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
enhancer of zeste 2 polycomb repressive complex 2 subunit/Histone-lysine N-methyltransferase EZH2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2189110] [GtoPdb: 2654] [UniProtKB: Q15910]
ChEMBL Inhibition of EZH2 (unknown origin) B 4.19 pIC50 65000 nM IC50 Bioorg Med Chem Lett (2016) 26: 4518-4522 [PMID:27485386]
ChEMBL Inhibition of recombinant human EZH2 using core histone as substrate by hotspot assay B 4.65 pIC50 22600 nM IC50 Eur J Med Chem (2020) 189: 112023-112023 [PMID:31978781]
ChEMBL Inhibition Assay: Test compounds were serially diluted 3-fold in DMSO in a 10 point-curve and 1 uL was spotted into a 384-well microplate in duplicate using a Platemate Plus equipped with 384-channel head (Thermo Scientific). The final top concentration of test compounds in the assay was 10 uM. Positive control (100% inhibition standard) was 1 mM final concentration of SAH and negative control (0% inhibition standard) contained 1 uL of DMSO. Test compounds were then incubated for 30 minutes with 40 uL per well of wild-type EZH2 (final concentration was 4 nM), Y641F EZH2 (final concentration was 0.1 nM) and A677G and A687V EZH2 (for each, final concentration was 2nM) and peptide in 1x assay buffer (20 mM BICINE pH =7.6, 1 mM DTT, 0.002% TWEEN 20 (generic: polysorbate 20), 0.005% BSG). For the wild-type EZH2 and A677G EZH2 assays, biotinylated peptide H3:21-44 with unmethylated K27 was present at a final concentration of 200 nM, while in the A687V EZH2 assay, biotinylated peptide H3. B 4.78 pIC50 16619.3 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition of N-terminally FLAG-tagged wild type EZH2 in EZH2/SUZ12/EED/RbAp48 complex (unknown origin) expressed in baculovirus infected in SF9 cells assessed as inhibition of methylation of nucleosomes at H3K27 by scintillation counting in presence of [3H]SAM B 4.96 pIC50 11000 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
ChEMBL Inhibition Assay: Test compounds were serially diluted 3-fold in DMSO in a 10 point-curve and 1 uL was spotted into a 384-well microplate in duplicate using a Platemate Plus equipped with 384-channel head (Thermo Scientific). The final top concentration of test compounds in the assay was 10 uM. Positive control (100% inhibition standard) was 1 mM final concentration of SAH and negative control (0% inhibition standard) contained 1 uL of DMSO. Test compounds were then incubated for 30 minutes with 40 uL per well of wild-type EZH2 (final concentration was 4 nM), Y641F EZH2 (final concentration was 0.1 nM) and A677G and A687V EZH2 (for each, final concentration was 2nM) and peptide in 1x assay buffer (20 mM BICINE pH =7.6, 1 mM DTT, 0.002% TWEEN 20 (generic: polysorbate 20), 0.005% BSG). For the wild-type EZH2 and A677G EZH2 assays, biotinylated peptide H3:21-44 with unmethylated K27 was present at a final concentration of 200 nM, while in the A687V EZH2 assay, biotinylated peptide H3. B 5.16 pIC50 6908.2 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition Assay: Test compounds were serially diluted 3-fold in DMSO in a 10 point-curve and 1 uL was spotted into a 384-well microplate in duplicate using a Platemate Plus equipped with 384-channel head (Thermo Scientific). The final top concentration of test compounds in the assay was 10 uM. Positive control (100% inhibition standard) was 1 mM final concentration of SAH and negative control (0% inhibition standard) contained 1 uL of DMSO. Test compounds were then incubated for 30 minutes with 40 uL per well of wild-type EZH2 (final concentration was 4 nM), Y641F EZH2 (final concentration was 0.1 nM) and A677G and A687V EZH2 (for each, final concentration was 2nM) and peptide in 1x assay buffer (20 mM BICINE pH =7.6, 1 mM DTT, 0.002% TWEEN 20 (generic: polysorbate 20), 0.005% BSG). For the wild-type EZH2 and A677G EZH2 assays, biotinylated peptide H3:21-44 with unmethylated K27 was present at a final concentration of 200 nM, while in the A687V EZH2 assay, biotinylated peptide H3. B 5.2 pIC50 6237.9 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition Assay: Test compounds were serially diluted 3-fold in DMSO in a 10 point-curve and 1 uL was spotted into a 384-well microplate in duplicate using a Platemate Plus equipped with 384-channel head (Thermo Scientific). The final top concentration of test compounds in the assay was 10 uM. Positive control (100% inhibition standard) was 1 mM final concentration of SAH and negative control (0% inhibition standard) contained 1 uL of DMSO. Test compounds were then incubated for 30 minutes with 40 uL per well of wild-type EZH2 (final concentration was 4 nM), Y641F EZH2 (final concentration was 0.1 nM) and A677G and A687V EZH2 (for each, final concentration was 2nM) and peptide in 1x assay buffer (20 mM BICINE pH =7.6, 1 mM DTT, 0.002% TWEEN 20 (generic: polysorbate 20), 0.005% BSG). For the wild-type EZH2 and A677G EZH2 assays, biotinylated peptide H3:21-44 with unmethylated K27 was present at a final concentration of 200 nM, while in the A687V EZH2 assay, biotinylated peptide H3. B 5.23 pIC50 5903.4 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition Assay: Compound 75 was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. Compound 75 was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained 5-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 5.32 pIC50 4800 nM IC50 US-8895245-B2. Inhibitors of human EZH2 and methods of use thereof (2014)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 5.32 pIC50 4800 nM IC50 US-9333217-B2. Inhibitors of human EZH2, and methods of use thereof (2016)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% TWEEN® 20 (generic: polvsorbate 20), 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated FLASHPLATE™ (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TOPCOUNT NXT HTS (Perkin Elmer). B 5.32 pIC50 4800 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition of N-terminally FLAG-tagged EZH2 Y641N mutant in EZH2/SUZ12/EED/RbAp48 complex (unknown origin) expressed in baculovirus infected in SF9 cells assessed as inhibition of methylation of nucleosomes at H3K27 by scintillation counting in presence of [3H]SAM B 5.72 pIC50 1900 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.33 pIC50 467 nM IC50 US-9333217-B2. Inhibitors of human EZH2, and methods of use thereof (2016)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% TWEEN® 20 (generic: polvsorbate 20), 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated FLASHPLATE™ (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TOPCOUNT NXT HTS (Perkin Elmer). B 6.33 pIC50 467 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition Assay: Compound 75 was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. Compound 75 was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained 5-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.33 pIC50 467 nM IC50 US-8895245-B2. Inhibitors of human EZH2 and methods of use thereof (2014)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% TWEEN® 20 (generic: polvsorbate 20), 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated FLASHPLATE™ (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TOPCOUNT NXT HTS (Perkin Elmer). B 6.42 pIC50 380 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition Assay: Compound 75 was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. Compound 75 was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained 5-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.42 pIC50 380 nM IC50 US-8895245-B2. Inhibitors of human EZH2 and methods of use thereof (2014)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.42 pIC50 380 nM IC50 US-9333217-B2. Inhibitors of human EZH2, and methods of use thereof (2016)
ChEMBL Inhibition Assay: Compound 75 was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. Compound 75 was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained 5-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.55 pIC50 283 nM IC50 US-8895245-B2. Inhibitors of human EZH2 and methods of use thereof (2014)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% TWEEN® 20 (generic: polvsorbate 20), 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated FLASHPLATE™ (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TOPCOUNT NXT HTS (Perkin Elmer). B 6.55 pIC50 283 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.55 pIC50 283 nM IC50 US-9333217-B2. Inhibitors of human EZH2, and methods of use thereof (2016)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.58 pIC50 263 nM IC50 US-9333217-B2. Inhibitors of human EZH2, and methods of use thereof (2016)
ChEMBL Inhibition Assay: Compound 75 was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. Compound 75 was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% Tween 20, 0.005% BSG). A substrate mix at 10 μL per well was added which contained 5-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated Flashplate (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TopCount NXT HTS (Perkin Elmer). B 6.58 pIC50 263 nM IC50 US-8895245-B2. Inhibitors of human EZH2 and methods of use thereof (2014)
ChEMBL Inhibition Assay: S-Adenosyl-L-homocysteine (SAH) was serially diluted 3 fold in DMSO for 10 points and 1 μL was plated in a 384 well microtiter plate. Positive control (100% inhibition standard) was 100 μM final concentration of SAH and negative control (0% inhibition standard) contained 1 μL of DMSO. SAH was then incubated for 30 minutes with 40 μL per well of EZH2 wild-type and mutants at 8 nM in pH 7.6 assay buffer (20 mM BICINE, 100 mM KCl, 1 mM DTT, 0.002% TWEEN® 20 (generic: polvsorbate 20), 0.005% BSG). A substrate mix at 10 μL per well was added which contained S-adenosylmethionine-Cl (SAM) at 150 nM and tritiated SAM at 100 nM, and biotinylated oligonucleosome at 150 nM in pH 7.6 assay buffer. Quenched enzyme reaction was transferred to a streptavidin-coated FLASHPLATE™ (Perkin Elmer, catalog number SMP410), allowed to bind for one hour, and detected on a TOPCOUNT NXT HTS (Perkin Elmer). B 6.58 pIC50 263 nM IC50 US-9175331-B2. Inhibitors of human EZH2, and methods of use thereof (2015)
DOT1 like histone lysine methyltransferase/Histone-lysine N-methyltransferase, H3 lysine-79 specific in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1795117] [GtoPdb: 2650] [UniProtKB: Q8TEK3]
ChEMBL Binding affinity at human recombinant DOT1L catalytic domain amino acid (1 to 472) by isothermal titration calorimetric assay B 6.44 pKd 360 nM Kd J Med Chem (2012) 55: 8066-8074 [PMID:22924785]
ChEMBL Binding affinity at human recombinant DOT1L catalytic domain amino acid (1 to 472)-nucleosome complex by isothermal titration calorimetric assay B 6.82 pKd 150 nM Kd J Med Chem (2012) 55: 8066-8074 [PMID:22924785]
ChEMBL Inhibition of DOT1-like Histone H3 Methyltransferase (unknown origin) B 6.57 pKi 270 nM Ki J Med Chem (2019) 62: 10005-10025 [PMID:31188592]
ChEMBL Inhibition of human recombinant DOT1L catalytic domain amino acid (1 to 472) using [3H]-SAM after 30 mins by scintillation counter B 6.8 pKi 160 nM Ki J Med Chem (2012) 55: 8066-8074 [PMID:22924785]
ChEMBL Competitive inhibition of recombinant human DOT1L using adenosine/deazaadenosine as substrate and SAM cofactor B 6.8 pKi 160 nM Ki J Med Chem (2013) 56: 8972-8983 [PMID:23879463]
ChEMBL Competitive inhibition of human recombinant DOT1L (1 to 420 amino acid residues) overexpressed in Escherichia coli BL21 (DE3) using [3H]-SAM as substrate assessed as inhibition of nucleosome methylation incubated for 30 mins prior to substrate addition measured after 1 hr by scintillation counting analysis B 6.22 pIC50 600 nM IC50 Bioorg Med Chem (2013) 21: 1787-1794 [PMID:23433670]
ChEMBL Competitive inhibition of human recombinant DOT1L (1 to 472 amino acid residues) expressed in Escherichia coli BL21 (DE3) using [3H]-SAM assessed as inhibition of nucleosome methylation incubated for 10 mins prior to substrate addition measured after 30 mins by scintillation counting analysis B 6.22 pIC50 600 nM IC50 Bioorg Med Chem (2013) 21: 1787-1794 [PMID:23433670]
ChEMBL Inhibition of human recombinant DOT1L (1 to 420 amino acids) expressed in Escherichia coli B 6.66 pIC50 220 nM IC50 Bioorg Med Chem Lett (2016) 26: 4518-4522 [PMID:27485386]
euchromatic histone lysine methyltransferase 2/Histone-lysine N-methyltransferase, H3 lysine-9 specific 3 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL6032] [GtoPdb: 2652] [UniProtKB: Q96KQ7]
ChEMBL Inhibition of G9a (unknown origin) B 6.24 pKi 570 nM Ki J Med Chem (2013) 56: 8972-8983 [PMID:23879463]
ChEMBL Inhibition of G9a (unknown origin) B 5.39 pIC50 4100 nM IC50 Bioorg Med Chem Lett (2016) 26: 4518-4522 [PMID:27485386]
ChEMBL Inhibition of G9a (unknown origin) by HMT assay B 5.7 pIC50 2000 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
ChEMBL Inhibition of recombinant human G9a using histone H3 (1 to 21) as substrate by hotspot assay B 5.74 pIC50 1820 nM IC50 Eur J Med Chem (2020) 189: 112023-112023 [PMID:31978781]
euchromatic histone lysine methyltransferase 1/Histone-lysine N-methyltransferase, H3 lysine-9 specific 5 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL6031] [GtoPdb: 2651] [UniProtKB: Q9H9B1]
ChEMBL Inhibition of EHMT1 (unknown origin) by HMT assay B 6.64 pIC50 230 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
lysine methyltransferase 2A/Histone-lysine N-methyltransferase MLL in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1293299] [GtoPdb: 2688] [UniProtKB: Q03164]
ChEMBL Inhibition of MLL (unknown origin) by HMT assay B 5.64 pIC50 2300 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
nuclear receptor binding SET domain protein 2/Histone-lysine N-methyltransferase NSD2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL3108645] [GtoPdb: 3220] [UniProtKB: O96028]
ChEMBL Inhibition of recombinant human N-terminal GST-tagged NSD2 CD (941 to 1240 residues) expressed in Escherichia coli using SAM as substrate in presence of nucleosomes by AlphaLISA method B 4.41 pIC50 39100 nM IC50 Eur J Med Chem (2021) 222: 113592-113592 [PMID:34147909]
SET domain containing 7, histone lysine methyltransferase/Histone-lysine N-methyltransferase SETD7 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL5523] [GtoPdb: 2703] [UniProtKB: Q8WTS6]
ChEMBL Inhibition of SETD7 (unknown origin) B 4.33 pIC50 47000 nM IC50 Bioorg Med Chem Lett (2016) 26: 4518-4522 [PMID:27485386]
ChEMBL Inhibition of SETD7 (unknown origin) by HMT assay B 4.52 pIC50 30000 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
SET domain bifurcated histone lysine methyltransferase 1/Histone-lysine N-methyltransferase SETDB1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2321646] [GtoPdb: 2705] [UniProtKB: Q15047]
ChEMBL Inhibition of SETDB1 (unknown origin) by HMT assay B 5.66 pIC50 2200 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
SUV39H1 histone lysine methyltransferase/Histone-lysine N-methyltransferase SUV39H1 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1795118] [GtoPdb: 2715] [UniProtKB: O43463]
ChEMBL Inhibition of SUV39H1 B 5.31 pKi 4900 nM Ki J Med Chem (2012) 55: 8066-8074 [PMID:22924785]
ChEMBL Inhibition of SUV39H1 (unknown origin) by HMT assay B 5.92 pIC50 1200 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
SUV39H2 histone lysine methyltransferase/Histone-lysine N-methyltransferase SUV39H2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL1795177] [GtoPdb: 2716] [UniProtKB: Q9H5I1]
ChEMBL Inhibition of SUV39H2 (unknown origin) by HMT assay B 4.49 pIC50 32000 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
ChEMBL Inhibition of SUV39H2 (unknown origin) B 4.66 pIC50 22000 nM IC50 Bioorg Med Chem Lett (2016) 26: 4518-4522 [PMID:27485386]
lysine methyltransferase 5C/Histone-lysine N-methyltransferase SUV420H2 in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2321644] [GtoPdb: 2718] [UniProtKB: Q86Y97]
ChEMBL Inhibition of SUV420H2 (unknown origin) by HMT assay B 5 pIC50 10000 nM IC50 Bioorg Med Chem Lett (2015) 25: 1532-1537 [PMID:25746813]
Indolethylamine N-methyltransferase in Human (target type: SINGLE PROTEIN) [ChEMBL: CHEMBL2131] [UniProtKB: O95050]
ChEMBL Inhibitory constant towards indole N-methyl-transferase B 5.7 pKi 2000 nM Ki J Med Chem (1983) 26: 1470-1477 [PMID:6620306]