PHA-680632 is potent inhibitor of Aurora A, Aurora B and Aurora C with IC50 of 27 nM, 135 nM and 120 nM, respectively. It has 10- to 200-fold higher IC50 for FGFR1, FLT3, LCK, PLK1, STLK2, and VEGFR2/3.

Aurora A:27 nM, Aurora B:135 nM, Aurora C:120 nM

PHA-680632对所有三种Aurora激酶（A、B和C）具有强大的抑制作用，其IC50值分别为27、135和120 nM。PHA-680632选择性抑制Aurora激酶，对FGFR1、FLT3、LCK、PLK1、STLK2、VEGFR2和VEGFR3的IC50值高出10至200倍，对另外22种激酶的IC50值高于10 μM。PHA-680632在广泛的细胞类型中显示出强大的抗增殖效果，其IC50值为0.06–7.15 μM，包括HeLa、HCT116、HT29、LOVO、DU145和NHDF细胞。PHA-680632（0.5 μM）会在肿瘤细胞中引起多倍体。PHA-680632的作用机制与抑制Aurora激酶一致。[1] PHA680632与辐射联合使用在癌细胞中产生加成效应，特别是在p53缺陷细胞中。联合使用离子辐射（IR）和PHA680632（100–400 nM）处理，然后进行IR，可在具有p53缺陷的HCT116细胞中，仅增强辐射诱导的Annexin V阳性细胞、微核形成和Brca1焦点形成，而不影响p53野生型对应物。[2]

HA-680632 (15–60 mg/kg) 在小鼠异种移植模型中，通过减少肿瘤细胞增殖并增加凋亡，抑制了HL60、A2780和HCT116细胞的肿瘤生长。PHA-680632 (45 mg/kg) 抑制了小鼠乳腺瘤病毒v-Ha-ras转基因小鼠中活化ras驱动的乳腺肿瘤生长，导致肿瘤完全稳定和部分回归。[1]

Aurora Kinase Inhibition Assay: Inhibition of kinase activity by PHA-680632 is assessed using a scintillation proximity assay format. The biotinylated substrate is transphosphorylated by the kinase in presence of ATP traced with γ33-ATP. The phosphorylated substrate is then captured using streptavidin-coated scintillation proximity assay beads and the extent of phosphorylation is evaluated by β-counter after a 4-hour rest for the floatation of the beads on a dense 5 M CsCl solution. In particular, a peptide derived from the Chocktide sequence (LRRWSLGL) is used as substrate for Aurora A, whereas the optimized peptide Auroratide is employed for Aurora B and C. The assay is run in a robotized format on 96-well plates. The potency of the compound toward Aurora kinases is evaluated and IC50 values are determined.

Cells (5 × 103 to 1.5 × 104 per cm2) are seeded in 24-well plate. After 24 hours, plates are treated with PHA-680632 and incubated for 72 hours. At the end of incubation time, cells are detached from each plate and counted using a cell counter. IC50s are calculated using percentage of growth versus untreated cont(Only for Reference)