AZD4547 is an FGFR family inhibitor. AZD4547 selectively inhibits FGFR1 phosphorylation and suppresses cancer cell proliferation through inhibition of FGFR1 signaling, and is able to act on FGFR1 (IC50:0.2 nM), FGFR2 (IC50:2.5 nM), FGFR3 (IC50:1.8 nM) and FGFR4 (IC50:165 nM). IC50:165 nM).[2]

FGFR3:1.8 nM (cell free), FGFR2:2.5 nM (cell free), KDR:24 nM (cell free), FGFR1:0.9 nM (cell free)

方法：NRK-52E细胞用2.5,5,10,20μM AZD4547预处理1小时，然后暴露于0.5μg/ mLLPS指定时间，观察AZD4547对NRK-52E细胞中 LPS 诱导的炎症缺陷。
结果：AZD4547在NRK-52E细胞中以20μM的浓度显著抑制细胞增殖。[2]
方法：NRK-52E细胞用10μM AZD4547预处理1小时，然后暴露于0.5μg/ mL LPS指定时间，研究了 NRK-52E 细胞中促炎因子的水平。
结果：AZD4547通过减少NRK-52E细胞中的TAK1/NF-κB信号通路来减弱LPS诱导的炎症因子。[2]

方法：C57BL/6 小鼠使用LPS（15 mg/kg，腹膜内注射）并用AZD4547治疗（在LPS治疗前1小时灌胃40 mg/kg），研究 AZD4547 在 LPS 诱导的急性肾损伤中的肾保护作用以及测量了促凋亡基因Bax和抗凋亡基因Bcl-2研究AZD4547对AKI中保留肾功能的影响是否通过调节细胞凋亡来介导。
结果：AZD4547可以预防LPS相关的肾功能不全，细胞凋亡和小鼠肾脏炎症。[1]

Cells were treated with AZD4547 or control for 3 hours at 37°C and then stimulated with 10 ng/mL aFGF/bFGF and 10 μg/mL heparin for 20 minutes. Western blotting was conducted with standard SDS-PAGE procedures and antibody incubation carried out overnight at 4°C. Secondary antibodies were applied and immunoreactive proteins visualized with Chemiluminescence substrate according to the manufacturer's instructions [1].

Cell lines were incubated with fixed concentrations of AZD4547 for 72 hours. For fluorescence-activated cell sorting (FACS), cells were fixed with 70% ethanol and then incubated with propidium iodide/RNase A labeling solution. Cell-cycle profiles were assessed with a FACSCalibur instrument and CellQuest analysis software. For apoptotic analysis, cells and media were gently harvested and centrifuged, followed by washing of cell pellets. Cells were then processed for Annexin V-fluorescein isothiocyanate (FITC) staining and propidium iodide uptake according to the manufacturer's instructions. The proportion of cells staining positive for Annexin V were then assessed with a FACSCalibur instrument and quadrant sorting was done by CellQuest analysis software [1].

Tumor xenografts were established by s.c. injection into the left flank with 0.1 mL tumor cells (1 × 10^6 for LoVo, 1 × 10^7 for HCT-15, and 1 × 10^7 for Calu-6) or 0.2 mL (2 × 10^7 for KMS11 and KG1a) mixed 1:1 with Matrigel, with the exception of LoVo and HCT-15, which did not include Matrigel. Mice were randomized into control and treatment groups when tumors reached the determined size of more than 0.2 cm3. Tumor volume (measured by caliper), animal body weight, and tumor condition were recorded twice weekly for the duration of the study. Tumor volume was calculated as described previously [1].