Derazantinib (ARQ-087) is a potent, ATP-competitive, orally active tyrosine kinase inhibitor with IC50 values ​​of 4.5/1.8/4.35/3.4 nM for chondrocyte FGFR1/FGFR2/FGFR3/FGFR4, respectively.

FGFR1:4.5 nM, FGFR3:4.5 nM, FGFR2:1.8 nM

方法：Derazantinib (ARQ-087)（0.1,0.3，1.1,3.3，10μM，2小时）处理异位表达 FGFR1、FGFR2、FGFR3 或 FGFR4 的 COS-1 细胞，用 100 pM FGF1/2/7 刺激 15 分钟，通过Western blot分析评估总FGFR和磷酸化FGFR。
结果：Derazantinib与EC一起抑制FGFR1、FGFR2、FGFR3和FGFR4的磷酸化IC50值分别为＜0.123μM，0.185μM，0.463μM和＞10μM。[1]

方法：Derazantinib (ARQ-087)（25,50,75 mg/kg）处理 SNU-16和NCI-H716异种移植无胸腺小鼠模型；Derazantinib （50,100,150mg/kg）处理Ba/F3-FGFR2异种移植无胸腺小鼠模型；Derazantinib （75mg/kg）处理Ba/F3-INSR异种移植无胸腺小鼠模型；均采用口服的方式并评估评估 Derazantinib 的体内抗肿瘤作用。
结果：Derazantinib在 Ba/F3-FGFR2 模型中显示出有效的肿瘤生长抑制作用，而未能抑制 Ba/F3-INSR 模型的生长；在SNU-16异种移植研究中，用75 mg/kg和50 mg/kg处理分别达到83%和69%的TGI；在NCI-H716人盲肠模型中， 50 mg/kg 和 75 mg/kg 分别显示出显著的 TGI，分别为 68%和 96%。 [1]

Derazantinib is titrated in DMSO utilizing a 3-fold dilution scheme and then diluted 10-fold further in deionized water for a final DMSO concentration of 10%. A volume (2.5 μL) of these dilutions or vehicle is added to each well of a reaction plate. FGFR1 or FGFR2 is added to assay buffer to each well in a volume of 17.5 μL for a final concentration of 0.50 or 0.25 nM, respectively. After a 30-minute pre-incubation period, ATP and substrate are added in assay buffer (5 μL) for final concentrations of 0-1,000 μM ATP and 80 nM biotinylated-PYK2, for a final reaction volume of 25 μL. The plates are incubated for 60 minutes at room temperature and then stopped in the dark by the addition of 10 μL stop/detection mixture prepared in assay buffer containing EDTA [1].

Cells are seeded at 3000-5000 cells per well with 130 μL media in 96-well tissue culture-treated plates. The cells are incubated overnight and subsequently treated with 3-fold serial dilutions of Derazantinib starting at 100 μM. The cells are returned to a 37°C humidified incubator for 72 hours. Cell proliferation is measured using the MTS assay [1].

Female NCr nu/nu mice (SNU-16) or CB17 SCID mice (NCI-H716) with well established (400 mg) subcutaneous tumors are given a single oral dose of Derazantinib or vehicle control. Plasma and tumor samples are collected 4 hours post single dose. Derazantinib is administered orally. The dosing volume for all groups is 10 mL/kg or 0.1 mL/10 g body weight [1].