GNF-7 is Bcr-Abl WT and Bcr-Abl T315I inhibitor with IC50 of 133 nM and 61 nM, respectively.

BCR-ABL:61 nM, BCR-ABL (WT):133 nM

GNF-7对野生型及突变型Bcr-Abl Ba/F3细胞表现出强大的抗增殖活性，其IC50小于11nM。在人类结肠癌细胞（Colo205和SW620）中，GNF-7同样展现出优异的生长抑制活性，IC50分别为5nM和1nM。[1] GNF-7强效且选择性地通过ACK1/AKT和GCK的联合抑制，抑制NRAS依赖型的急性髓性白血病和急性淋巴细胞性白血病细胞。[2]

GNF-7在小鼠中展示出卓越的药代动力学参数。在使用转化的T315I-Bcr-Abl-Ba/F3细胞系的发光异种移植小鼠模型中，GNF-7（10 mg/kg，口服）有效抑制肿瘤生长。[1] 在携带人类突变NRAS表达的MOLT-3-luc+肿瘤的NSG小鼠中，GNF-7（15 mg/kg，口服）显著降低疾病负担，延长总生存期，并强烈抑制phospho-AKT与phospho-RPS6。[2]

For kinase assays, purified CDC5L(295-795)-His6 is mixed with [γ-32P]ATP, COS-7 cell extract, and incubated in 100 μL 20 mM HEPES, pH 7.5, 50 mM NaCl, 2 mM MnCl2, 10 mM MgCl2, 0.5% NP-40, 0.5 mM PMSF, 5 mM benzamidine hydrochloride, 5 mM NaF, 1 mM NaVO3 and the specific inhibitor at 30°C for 10 minutes. Cell extract as a source of kinase activity is prepared from subconfluent, serum-stimulated COS-7 cells lysed in 20 mM HEPES-NaOH, pH 7.5, 50 mM NaCl, 1% Triton X-100, 10% glycerol, protease and phosphotase inhibitors. Phosphorylated proteins are separated by electrophoresis in 15% polyacrylamide-SDS gels. Specific inhibitors included 20 μM staurosporine, 10 μM genistein, 1 μM CVT-313, 10 μM Rp-MB-cAMPS and 50 μM PD98059[1].