ENMD2076是一种多靶点激酶抑制剂，抑制AuRORa A、FLT3、KDR/VEGFR2、FLT4/VEGFR3、FGFR1、FGFR2、Src、PDGFRα的IC50值分别为1.86、14、58.2、15.9、92.7、70.8、20.2 and 56.4 nM。

ENMD-2076, a multi-targeted kinase inhibitor, has specific activity against Aurora A, Flt3, KDR/VEGFR2, Flt4/VEGFR3, FGFR1, FGFR2, Src, PDGFRα.

在MDA-MB-231异种移植模型中, ENMD-2076可以抑制形成新血管,同时使形成的血管发生退化.在HT29异种移植模型中,ENMD-2076对Flt3以及VEGFR2 / KDR和FGFR1 / 2的激活持续发挥抑制作用.在H929人浆细胞瘤异种移植物中,口服ENMD-2076（50-200 mg / kg /day）,其中磷酸 - 组蛋白3（pH3）,Ki-67显著降低,裂解的胱天蛋白酶-3显著增加.

ENMD-2076作用于PI3K/AKT通路,下调凋亡抑制蛋白。ENMD-2076能够抑制aurora A和B激酶,且诱导细胞周期停在G2/M 期。在多种作用于血管生成的激酶中（IC50=1.86-120 nM）,ENMD-2076能够发挥作用,比如VEGFR2/KDR 和VEGFR3, FGFR1和FGFR2, 及PDGFRα。在多种人类实体瘤和血癌细胞系中（IC50=0.025 -0.7 μM）,ENMD-2076使细胞停滞在G2/M 期,诱导细胞凋亡

Kinase assays: Recombinant Aurora A and B kinase enzymes and appropriate PanVera Z'-Lyte kinase assay kits are purchased. Assays are carried out in kinase assay buffer (50 mM of HEPES, pH 7.5, 10 mM of MgCl2, 5 mM of EGTA, 0.05% Brij-35) supplemented with 2 mM of DTT. Activities are determined at an ATP concentration equivalent to the apparent Km for each enzyme, and an enzyme concentration that results in approximately 30% phosphorylation of the peptide substrate after 1 hour. Dose–response curves of relative enzyme activity versus ENMD-2076 concentration are plotted with Grafit and used to calculate IC50 values. Potency of ENMD-2076 free base against a select panel of 100 kinase enzymes is determined using the SelectScreen kinase profiling service. ATP concentrations are at the apparent Km for each enzyme or 100 μM if the apparent Km could not be reached. Percent inhibition is determined at an ENMD-2076 free base concentration of 1 μM; for kinases where significant inhibition is noted, IC50 values are determined by generating full 10-point dose–response curves.

The antiproliferative effect of ENMD-2076 on adherent tumor cell lines is measured by plating 500 cells per well in a 96-well plate and incubating with ENMD-2076 for 96 hours. Cellular proliferation is measured using the sulforhodamine B assay. The leukemia-derived, nonadherent cell lines are assayed by plating 5 × 103 cells per well in a 96-well plate. The cells are incubated with ENMD-2076 for 48 hours and then survival is assayed using the Alamar Blue reagent. To measure the effect of ENMD-2076 on VEGF- and fibroblast growth factor (FGF)-induced proliferation of human umbilical vein endothelial cell (HUVEC), cells are serum starved for 6 hours, then treated with ENMD-2076 free base, and stimulated with 5 ng/mL bFGF or 25 ng/mL VEGF (R and D Systems) for 72 hours. Cell proliferation is measured using WST-(Only for Reference)

934353-76-1

C21H25N7

375.48

Ethanol:<1 mgml
DMSO:97 mg/mL (258.3 mM)
H2O:1 mg/mL (2.66 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years