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

METHODS: COS-1 cells ectopically expressing FGFR1, FGFR2, FGFR3, or FGFR4 were treated with derazantinib (ARQ-087) (0.1, 0.3, 1.1, 3.3, 10 μM, 2 hours) and stimulated with 100 pM FGF1/2/7 for 15 minutes. Total and phosphorylated FGFRs were assessed by Western blot analysis.
RESULTS Derazantinib inhibited phosphorylation of FGFR1, FGFR2, FGFR3, and FGFR4 with IC50 values ​​of <0.123 μM, 0.185 μM, 0.463 μM, and >10 μM, respectively, together with EC. [1]

METHODS: Derazantinib (ARQ-087) (25, 50, 75 mg/kg) treated SNU-16 and NCI-H716 xenograft athymic mouse models; Derazantinib (50, 100, 150 mg/kg) treated Ba/F3-FGFR2 xenograft athymic mouse models Thymic mouse model; Derazantinib (75 mg/kg) treated Ba/F3-INSR xenograft athymic mouse model; both were administered orally and the in vivo anti-tumor effect of Derazantinib was evaluated.
RESULTS Derazantinib showed potent tumor growth inhibition in the Ba/F3-FGFR2 model but failed to inhibit the growth of the Ba/F3-INSR model; in the SNU-16 xenograft study, 75 mg/kg and 50 mg /kg treatment achieved TGI of 83% and 69%, respectively; in the NCI-H716 human cecum model, 50 mg/kg and 75 mg/kg showed significant TGI of 68% and 96%, respectively. [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].