ODM-203, a Selective Inhibitor of FGFR and VEGFR, Shows Strong Antitumor Activity, and Induces Antitumor Immunity

FGFR2:16nM, VEGFR1:26nM, VEGFR3:5 nM, FGFR3:6nM, FGFR1:1nM, FGFR4:35 nM, VEGFR2:9nM

ODM-203 在生化实验中选择性地并且具有相同的效力抑制 FGFR 和 VEGFR 家族激酶，在低nM范围内显示出等效的抑制效果（IC50 6-35 nmol/L）。在细胞实验中，ODM-203 抑制 VEGFR 引导的管状形成（IC50 33 nmol/L），其抑制效力与它在 FGFR 依赖的细胞系中抑制增殖的效力相似（IC50 50-150 nmol/L）。

在体内，ODM-203 在 FGFR 依赖的异种移植模型和血管生成异种移植模型中均展现出了强大的抗肿瘤活性，且在相似的、良好耐受的剂量下实现。

Inhibition of FRS2 Tyrosine 196 phosphorylation by ODM-203 in FGFR-dependent cell lines was measured using an MSD 96-well multiarray Phospho-FRS2 Tyr196 assay (MesoScale Diagnostics) .?Briefly, the cell lines were seeded at a density of 75,000 cells/well on poly-d-lysine-coated 96-well plates?in the cell culture media .?The cells were allowed to attach overnight and subsequently treated with the vehicle (0.5% DMSO) or increasing concentrations of ODM-203 for 20 minutes.?The cell culture media were aspirated and the cells lysed in MSD Tris Lysis Buffer?supplemented with 10 mmol/L NaF, 1× phosphatase inhibitor cocktail 2 and 3 and Complete Protease Inhibitor Cocktail .?The electrochemiluminescence signal was detected with a SECTOR Imager 2400 plate reader coupled to a CCD camera.?Data were expressed as percentages of vehicle control values and analyzed with GrapPadPrism 7.03 .?Each test concentration was studied at least in triplicate and inhibition percentages were calculated for the parallel samples.?Average IC50 values were calculated from two independent experiments.

Athymic Nude-Foxn1nu female mice (9 weeks old;?Harlan, the Netherlands) were subcutaneously injected with 1 million H1581, KMS11, RT4, or SNU16 cells in 100 μL of McCoy's 5a modified medium and Matrigel (BD) (1:1).?Tumor growth was monitored twice weekly by caliper measurements.?Oral treatment with ODM-203 and AZD-4547 was started when the average tumor volume reached 100 mm^3 and continued for 21 days for the RT4 xenograft model (n = 12/group) and 12 days for the SNU16 xenograft model (n = 6/group).?Necropsy, and plasma and tumor sampling were carried out 4 hours after the last dosing.Doses for 12.5 mg/kg AZD4547 and 40 mg/kg sorafenib were chosen based on published data.?Oral treatment (ODM-203 or AZD4547) was initiated when the average tumor volume reached ≈125 mm^3.?Mean tumor volumes were calculated for each treatment group.