Dorsomorphin (BML-275) is an AMPK inhibitor (Ki=109 nM) that is selective and ATP-competitive. Dorsomorphin inhibits the BMP type I receptors ALK2, ALK3, and ALK6. Dorsomorphin induces autophagy, and possesses antitumor activity.

AMPK:109 nM(Ki)

方法：人肿瘤细胞 HeLa 和 HCT116 用 Dorsomorphin (1.25-80 μM) 处理 24 h，使用 CCK-8 方法检测细胞活力。
结果：Dorsomorphin 抑制 HeLa 和 HCT116 细胞的活力，IC50 值分别为 10.71 μM 和 11.34 μM。[1]
方法：ATL 患者来源的 PBMCs 细胞用 Dorsomorphin (5-25 μM) 处理 24 h，使用 Flow Cytometry 检测细胞凋亡情况。
结果：Dorsomorphin 以剂量依赖的方式增加了急性和慢性型 ATL 患者 PBMC 中早期凋亡细胞的频率。[2]

方法：为检测体内抗肿瘤活性，将 Dorsomorphin (10 mg/kg) 腹腔注射给携带人类肿瘤 S1T 的 NOD/SCID 小鼠，每天一次，持续二十八天。
结果：Dorsomorphin 抑制了 NOD/SCID 小鼠中人 ATL 肿瘤异种移植物的生长。[2]
方法：为检测体内对 SMAD 活性的影响，将 Dorsomorphin (10 mg/kg) 单次腹腔注射给 iron-dextran 处理的 C57BL/6 小鼠。
结果：Dorsomorphin 消除了 iron-dextran 引起的铁介导的肝脏 SMAD1/5/8 磷酸化的增加。[3]

HT1080 cells are seeded in 24-well plates (2×104 cells per well) and treated with Dorsomorphin in the presence or absence of glucose or 10 mM 2DG for 2 h. HT1080 cells that overexpressed the wild-type and dominant negative AMPKα1 are prepared by transfecting plasmid DNA (pAMPKα1-wt, pAMPKα1-D168A and pcFlag as a control) in 6-well plates, seeding in 24-well plate and treating with UPR inhibitors. Cells are lysed with cell lysis buffer (20 mM Tris-HCl, pH 7.5, 250 mM NaCl, 10% glycerol, 0.5% NP-40, 1 mM EDTA, 1 mM EGTA, 0.2 mM PMSF, 1 μg/mL pepstatin, 0.5 μg/mL leupeptin, 5 mM NaF, 2 mM Na3Vo4, 2 mM β-glycerophosphate, 1 mM DTT). Relative AMPK kinase activity (mean±SD of duplicate determinations) to control sample (vehicle or pcFlag under normal growth conditions) is determined using the CycLex AMPK kinase assay kit[2].

Dorsomorphin is dissolved in DMSO (10 mM) and stored,and then diluted with appropriate media (DMSO 0.5%) before use[2].HeLa and 786-O cells are treated with various concentrations of Dorsomorphin (0,0.3,1,3,10 μM ),Versipelostatin and Phenformin in the presence or absence of 10 mM 2DG or 1 μg/mL of Tunicamycin as a stressor for 30 h in 96-well plates.For the combination study,786-O cells are treated with various concentrations of UPR inhibitors in the presence or absence of 10 mM 2DG for 24 h.The medium is then replaced with fresh growth medium,and cells are cultured for a further 15 h.Subsequently,MTT is added to the culture medium,and the absorbance of each well is determined.For the viability assay under glucose-withdrawal conditions,HT1080 cells are treated with various concentrations of Dorsomorphin and phenformin in 12-well plates in the presence or absence of glucose for 18 h,seeded in 96-well plates with growth medium,and then cultured for a further 48 h before MTT is added.Relative cell survival (mean±SD of quadruplicate determinations) is calculated by setting each control absorbance from untreated cells as 100%.The effects of drug combinations at concentrations producing 80% cell growth inhibition (IC80) are analyzed using the isobologram method[2].