Nintedanib 是一种有效的三重血管激酶抑制剂，能够抑制VEGFR1 (IC50:34 nM)、VEGFR2 (IC50:13 nM)、VEGFR3 (IC50:13 nM)，FGFR1 (IC50:69 nM)、FGFR2 (IC50:37 nM)、FGFR3 (IC50:108 nM)，PDGFRα (IC50:59 nM)、PDGFRβ (IC50:65 nM)。

Nintedanib is an inhibitor of the receptor tyrosine kinases VEGFR/FGFR/PDGFR (IC50s: 13-34/37-610/59/65 nM for VEGFR1-3, FGFR1-4, and PDGFRα/β, respectively).

Nintedanib (BIBF 1120) is an indolinone derivative potently blocking VEGFR, PDGFR, and FGFR kinase activity (IC50: 20-100 nmol/L). BIBF 1120 inhibits mitogen-activated protein kinase and Akt signaling pathways in three cell types contributing to angiogenesis, endothelial cells, pericytes, and smooth muscle cells, resulting in inhibition of cell proliferation (EC50: 10-80 nmol/L) and apoptosis [1]. Three human NPC cell lines, CNE-2, HNE-1, and HONE-1, were treated with BIBF 1120 (0–10 μM) for 72 hours. BIBF 1120 significantly inhibited the growth of these three cell lines in a dose-dependent manner (IC50s: 5.62 μM, 4.16 μM, and 6.32 μM) [2].

In all tumor models tested thus far, BIBF 1120 is highly active at well-tolerated doses (25-100 mg/kg daily p.o.), as measured by magnetic resonance imaging of tumor perfusion after 3 days, reducing vessel density and vessel integrity after 5 days, and inducing profound growth inhibition [1]. BIBF 1120 demonstrated significant growth inhibition of NPC tumors in the HNE-1 xenograft model in nude mice as a single agent. At the same time, BIBF 1120 showed greater inhibition of tumor growth in the human NPC cell line xenograft model in nude mice when combined with DDP than in the DDP single-agent group or the control group. Body weight reduction was observed in the DDP single-agent group and combination group, and temporary body weight reduction was observed in the BIBF 1120 single-agent group [2]. TFTD (150 mg/kg/day) and/or nintedanib (40 mg/kg/day) were orally administered to the mice twice daily from day 1 to day 14. The tumor growth inhibition with combination therapy was 61.5, 72.8, 67.6 and 67.5% for the DLD-1, DLD-1/5-FU, HT-29, and HCT116 xenografts, respectively. This was significantly (P<0.05) higher than the effects of monotherapy with either TFTD or nintedanib [3].

The cytoplasmic tyrosine kinase domain of VEGFR-2 (residues 797–1355 according to sequence deposited in databank SWISS-PROT P35968) was cloned into pFastBac fused to GST and extracted as described in supplementary methods. Enzyme activity was assayed using standard conditions using a random polymer (Glu/Tyr 4:1) and in the presence of 100 μmol/L ATP (for details, see supplementary methods). For all other kinase assays, the entire cytoplasmic domains of the receptors (from the end of the transmembrane to the COOH terminus) were cloned into pFastBac vector containing GST and assayed under standard conditions [1].

HUVEC, HUASMC, and BRP were cultured as described above. Two hours before the addition of ligands, BIBF 1120 was added to the cultures. Cell lysates were generated according to standard protocols. Western blotting was done using standard SDS-PAGE methods, loading 50 to 75 μg of protein per lane, with detection by enhanced chemiluminescence. Total and phosphorylated mitogen-activated protein kinase (MAPK) was analyzed using monoclonal antibodies M3807 and M8159. Total Akt was detected using the polyclonal antibody and phosphorylated Akt (Ser473) was analyzed with the monoclonal antibody. Cleaved caspase-3 was detected with the monoclonal antibody [1].

Five-week-old to 6-wk-old athymic NMRI-nu/nu female mice (21–31 g) were purchased from Harlan. After acclimatization, mice were inoculated with 1 to 5 × 10^6 (in 100 μL) FaDu, Caki-1, SKOV-3, H460, HT-29, or PAC-120 cells s.c. into the right flank of the animal. F344 Fischer rats after acclimatization were injected with 5 × 10^6 (in 100 μL) GS-9L cells s.c. into the right flank of the animal. For pharmacokinetic analysis, blood was isolated at indicated time points from the retroorbital plexus of mice and plasma was analyzed using high-performance liquid chromatography-mass spectrometry methodology [1].

656247-17-5

C31H33N5O4

539.636

BIBF 1120;Intedanib;尼达尼布

DMSO:6 mg/mL (11.11 mM)
H2O:<1 mgml
Ethanol:3 mg/mL (5.55 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years