UNC3866 _Histone Methyltransferase 抑制劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEUNC3866Cat. No.: HY-100832CAS No.: 1872382-47-2分式: CHNO分量: 795.02作靶點(diǎn): Histone Methyltransferase作通路: Epigenetics儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 27 mg/mL (33.96 mM)* means sol

2、uble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 1.2578 mL 6.2891 mL 12.5783 mL5 mM 0.2516 mL 1.2578 mL 2.5157 mL10 mM 0.1258 mL 0.6289 mL 1.2578 mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 UNC3866種有效的 CBX7-H3 拮抗劑，IC50 為 661.2 nM。IC50 & Target

3、IC50: 661.2 nM (CBX7) 1體外研究UNC3866, a potent antagonist of the methyl-lysine (Kme) reading function of the Polycomb CBX and CDYfamilies of chromodomains. UNC3866 binds the chromodomains of CBX4 and CBX7 most potently with a Kd1/2 Master of Small Molecules 您邊的抑制劑師www.MedChemEof 100 nM for each, and i

4、s 6- to 18-fold selective versus seven other CBX and CDY chromodomains whilebeing highly selective versus 250 other protein targets. UNC3866 inhibits PC3 cell proliferation, a knownCBX7 phenotype, while UNC4219, a methylated negative control compound, has negligible effects.UNC3866 is a potent and c

5、ellularly active antagonist of PRC1 chromodomains. UNC3866 is the most potentligand reported for CBX7 with a Kd of 972.4 nM. UNC3866 is equipotent for CBX4, which is most similar toCBX7, while it is 18-, 6- and 12-fold selective for CBX4/7 over CBX2, -6 and -8, respectively. Additionally,UNC3866 is

6、65-fold selective for CBX4/7 over CDY1 and 9-fold selective for CBX4/7 over CDYL1b andCDYL2 1.PROTOCOLKinase Assay 1 The effect of UNC3866 on the methyltransferase activity of G9a, EHMT1, SUV39H1, SUV39H2, SETDB1,SETD8, SUV420H1, SUV420H2, SETD7, MLL1 trimeric complex, MLL3 tetrameric complex, EZH2

7、trimericcomplex, PRMT1, PRMT3, PRMT5-MEP50 complex, PRMT6, PRMT7, PRMT8, PRDM9, SETD2, SMYD2,SMYD3, BCDIN3D and DNMT1 is assessed by monitoring the incorporation of tritium-labeled methyl groupto lysine or arginine residues of peptide substrates using Scintillation Proximity Assay (SPA). Assays arep

8、erformed in a 20 L reaction mixture containing 3H-SAM at substrate concentrations close to the Km valuesfor each enzyme. Three concentrations (1 M, 10 M, and 50 M) of UNC3866 are used in all selectivityassays. To stop the enzymatic reactions, 7.5 M Guanidine hydrochloride is added, followed by 180 L

9、 ofbuffer (20 mM Tris, pH 8.0). The reactions are mixed and then transferred to a 96-well FlashPlate. Thereaction mixtures in Flash plates are incubated for 1 hour and the CPM are measured using a TopCountplate reader. The CPM counts in the absence of compound for each data set are defined as 100% a

10、ctivity. Inthe absence of the enzyme, the CPM counts in each data set are defined as background (0%) 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 PC3 cells are seeded at 200 cells/well into 24-well plates. Cells are allowed to adhere o

11、vernight. The media(DMEM supplemented with 10 % FBS) is then exchanged with fresh media containing DMSO, UNC3866 orUNC4219. On day three, the media are exchanged with fresh media containing DMSO, UNC3866 orUNC4219. For dose-response studies, the EC50 is derived from a six-point titration ranging fro

12、m 100 M to0.4 M of UNC3866 or UNC4219. At day 0, 3 or 6, cells are fixed with ice-cold methanol for 30 sec. andrehydrated with PBS. Nuclei of the cells are stained with DAPI (0.05 g/mL) and numerated using HighContent Microscopy. For dose-response studies, the cell count of UNC3866- or UNC4219-treat

13、ed cells isnormalized to the average cell count of DMSO-treated cells. The EC50 is calculated using the “l(fā)oginhibitorvs. the normalized response-Variable slope” equation in GraphPad Prism 5 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Stuckey JI, et al. A cellular chemical probe targeting the chromodomains of Polycomb repressive complex 1. Nat Chem Biol. 2016Mar;12(3):180-7.M