SGC2085 is an effective and selective coactivator-associated arginine methyltransferase 1 (CARM1) inhibitor (IC50: 50 nM).

CARM1:50 nM

除PRMT6（IC50=5.2 μM）外，SGC2085不抑制其他PRMTs。在测试三种不同浓度（1, 10, 和50 μM）的21种人类蛋白质甲基转移酶中，SGC2085显示出完全的选择性。为了表征SGC2085在溶液中的作用机制，于不同浓度的SAM和肽底物下测定IC50值。底物肽或辅因子浓度的增加不影响IC50值，这表明抑制机制为非竞争性，这一点之前已在其他结合于底物口袋的蛋白质甲基转移酶抑制剂上显示过[1]。在HEK293细胞中以10 μM测试高达48小时时，SGC2085未观察到细胞活性，而双重CARM1/PRMT6抑制剂MS049的10 μM则能够阻断BAF155的甲基化。我们假设SGC2085缺乏细胞活性是由于其较差的渗透性[1]。

SGC2085 is dissolved in DMSO and diluted with the appropriate medium before use. HEK293 cells are grown in 12-well plates in DMEM supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (100 μg/mL). Thirty per cent confluent cells are treated with inhibitors or DMSO. After 48 h, media are removed and cells are lysed in 100 μL of total lysis buffer (20 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 10 mM MgCl2, 0.5% Triton X-100, 12.5 U/mL benzonase), complete EDTA-free protease inhibitor cocktail. After 3 min incubation at room temperature, SDS is added to 1% final concentration. Lysates are run on SDS-PAGE, and immunoblotting is done as outlined below to determine the levels of unmethylated and methylated BAF155[1].