Derazantinib (ARQ-087) (ARQ-087) is an ATP competitive tyrosine kinase inhibitor. It exhibits potent activity against FGFR1/FGFR2/FGFR3 chondrocytes (IC50s: 4.5 nM/1.8 nM/4.5 nM).

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

In cells, Derazantinib effectively inhibits FGFR2 auto-phosphorylation and the activity of proteins downstream in the FGFR pathway (FRS2α, AKT, ERK), showcasing its anti-proliferative effects on cell lines affected by FGFR dysregulation due to amplifications, fusions, and mutations. Studies reveal a clear correlation between Derazantinib's ability to induce G1 cell cycle arrest and promote apoptosis in cell lines with elevated FGFR2 protein. Additionally, Derazantinib counteracts FGF2-mediated growth arrest with an EC50 of approximately 100 nM, maintaining safety at concentrations up to 500 nM and significantly inhibiting FGF2 effects within a 70-500 nM range. Beyond its influence on cell proliferation, Derazantinib prevents FGF-mediated deterioration of the extracellular matrix and premature chondrocyte senescence, supporting chondrocyte differentiation in tibia cultures. Its inhibitory action is specific to FGFR1-4, distinguishing Derazantinib from other receptor tyrosine kinases inhibitors in cell-free kinase assays, and extends to FGFR1 and FGFR2 mutants linked to craniosynostoses.

Derazantinib effectively inhibits tumor growth in FGFR2-altered SNU-16 and NCI-H716 xenograft models with gene amplification and fusions [1]. Additionally, 81.3% of embryos injected with Derazantinib exhibit abnormal external phenotypes, likely due to inhibited proliferation of limb bud mesenchyme [2].

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].