FGFR4-IN-24 is a selective irreversible covalent inhibitor of FGFR4 with an IC50 of 1.2 nM. It demonstrates significantly weaker activity against the other three FGFR family kinases (FGFR1-3). FGFR4-IN-24 effectively inhibits the FGF19/FGFR4 signaling pathway, suppressing the proliferation of the HuH-7 hepatocellular carcinoma cell line (GI50= 17 nM). In HuH-7 mouse xenograft models, FGFR4-IN-24 shows notable antitumor activity. This compound is applicable for hepatocellular carcinoma research.

FGFR4:1.2 nM

FGFR4-IN-24 (Compound 10v) exhibits enhanced apparent inhibitory activity with increased pre-incubation time in accordance with its covalent modification ability of FGFR4. It demonstrates greater potency indicated by a k_inact/K_I of 4.8 × 10^5 M^-1 s^-1, due to tighter binding to the FGFR4 kinase (K_I = 0.62 nM), which increases non-covalent affinity. FGFR4-IN-24 inhibits HuH-7 cell line proliferation over 72 hours (IC_50 = 1.2 nM, GI_50 = 17 nM). At 1 μM, FGFR4-IN-24 significantly inhibits FGFR4 by 79%, showing high selectivity. Furthermore, at concentrations ranging from 1 to 1000 nM over 2 hours, it inhibits the phosphorylation of the FGFR4 substrate FRS2α and reduces phosphorylation levels of ERK in HuH-7 cells.

FGFR4-IN-24 (Compound 10v) administered at 25 and 50 mg/kg via intraperitoneal injection twice daily for 21 days inhibits tumor growth in a dose-dependent manner in the HuH-7 xenograft mouse model while maintaining body weight.