GNF-5是 Bcr-Abl 的特异性非 ATP 竞争性抑制剂，IC50为0.22uM。它是 GNF-2 的类似物，具有改进的药代动力学特性。

GNF-5 is a specific non-ATP competitive inhibitor of Bcr-Abl (IC50: 0.22±0.1 uM, Wild-type Abl). It is an analog of GNF-2 with improved pharmacokinetic properties.

GNF-5（75 mg/kg）与尼罗替尼（50 mg/kg）联用可使T315I Bcr-Abl BMT模型中总存活数提高.GNF-5（100 mg/kg）表现出对异种移植和骨髓移植模型中野生型和T315I Bcr-Abl依赖性增殖的疗效.

GNF-5的抗增殖活性较强,可抑制wt-Bcr-Abl（EC50：430 nM）和E255K突变型（EC50：580 nM）Bcr-Abl细胞增殖。GNF-5与尼罗替尼或伊马替尼联用可抑制体外耐药变异的出现,且在抗Bcr-Abl T315I突变体的细胞试验和生物化学中显示了额外的抑制活性。

Kinetic characterization of Abl inhibition: The ATP/NADH-coupled assay system in a 96-well format is used to determine the initial velocity of Abl tyrosine kinase catalyzed peptide phosphorylation. The reaction mixture contained 20 mM Tris-HCl, (pH 8.0), 50 mM NaCl, 10 mM MgCl2, 2 mM PEP [2-(Phosphonooxy)- 2-propenoic acid) and 20 μg Abl peptide substrate (EAIYAAPFAKKK), fixed or varied (to determine inhibitor kinetic parameters) concentration of inhibitor applied, 1/50 of the final reaction mixture volume of PK/LDH enzyme (pyruvate kinase/lactic dehydrogenase enzymes from rabbit muscle), 160 μM NADH, 0.16 μM Abl, and ATP added last to start the reaction. Absorbance data are collected every 20s at 340 nm using a SpectraMax M5 Microplate Reader.

Ba/F3.p210 cells are obtained by transfecting the IL-3-dependent murine hematopoietic Ba/F3 cell line with a pEYK vector containing p210BCR-ABL and Bcr-Abl mutations. All cell lines are cultured with 5% CO2 at 37 °C in RPMI 1640 with 10% fetal bovine serum (FBS) and supplemented with 1% l-glutamine. Parental Ba/F3 cells are similarly cultured with 10% WEHI-conditioned medium as a source of IL-3. Transfected cell lines are cultured in media supplemented with 25 μg/mL zeocin. The 48 h cell proliferation studies are obtained using the CellTiter-Glo assay.(Only for Reference)

778277-15-9

C20H17F3N4O3

418.376

GNF 5

Ethanol:16 mg/mL (38.2 mM)
DMSO:77 mg/mL (184 mM)
H2O:<1 mgml
Powder: -20°C for 3 years
In solvent: -80°C for 2 years