AEE788 (NVP-AEE 788) has been used in trials studying the treatment of Cancer, Glioblastoma Multiforme, and Brain and Central Nervous System Tumors.

EGFR:2 nM, HER2/ErbB2:6 nM, FLK1:59 nM, c-Fms:60 nM, c-ABL:52 nM

AEE788（50 mg/kg）抑制盲肠和腹膜肿瘤的生长（> 50％）,并将植入裸鼠盲肠的HT29细胞的淋巴结转移发生率降低至70％,对体重无影响.50 mg/kg AEE788作用于NeuT/ErbB2 GeMag模型诱导肿瘤消退达57%.AEE788有效抑制A431肿瘤中的EGF诱导的EGFR磷酸化和GeMag肿瘤中的erbB2磷酸化.AEE788在NCI-H596或DU145异种移植模型中产生对肿瘤生长的剂量依赖性抑制,仅有轻微的体重变化.AEE788作用于K562肿瘤细胞,可以促进LBH589调节的活性氧簇的产生,可以增强凋亡. AEE788剂量依赖性地抑制由VEGF诱导的血管生成并且不抑制bFGF诱导的血管生成.AEE788作用于HT29盲肠癌,明显降低pEGFR和pVEGFR 的表达水平,但是不改变EGF,VEGF,EGFR或VEGFR的表达水平.和CPT-11联用,AEE788明显抑制淋巴癌转移 AEE788抑制Daoy,DaoyPt和DaoyHER2移植瘤的生长,抑制分别达51%,45%和72%.

0.2-1.0 μM AEE788抑制HT29细胞中EGFR和Akt的磷酸化。AEE788抑制人皮肤SCC细胞系（Colo16,HaCaT,SRB1和SRB12细胞）中EGFR,VEGFR2,Akt和MAPK的磷酸化,导致生长抑制和诱导细胞凋亡。 AEE788有效抑制A431细胞中的EGFR磷酸化,IC5??0为11 nM。 AEE788还抑制CHO细胞中KDR的磷酸化和BT-474细胞中的erbB2的磷酸化,而对A31细胞中的PDGF-诱导的磷酸化没有任何影响。AEE788作用于髓母细胞瘤细胞系,抑制细胞增殖,抑制EGF和神经调节蛋白诱导的HER1,HER2和HER3激活。AEE788抑制NCI-H596,MK,BT-474和SK-BR-3细胞增殖,IC50分别为78,56,49和381 nM。AEE788也抑制EGF和VEGF促进的人脐静脉内皮细胞增殖,IC50分别为43和155 nM。

Protein Kinase Assays: The in vitro kinase assays are performed in 96-well plates (30 μL) at ambient temperature for 15–45 min using the recombinant glutathione S-transferase-fused kinase domains (4-100 ng, depending on specific activity). [γ33P]ATP is used as phosphate donor and polyGluTyr-(4:1) peptide as acceptor. With the exception of protein kinase C-α, cyclin-dependent kinase 1/cycB and protein kinase A are protamine sulfate (200 μg/mL), histone H1 (100 μg/mL), and the heptapeptide Leu-Arg-Arg-Ala-Ser-Leu-Gly (known as Kemptide Bachem) respectively and are used as peptide substrates. Assays are optimized for each kinase using the following ATP concentrations: 1.0 μM (c-Kit, c-Met, c-Fms, c-Raf-1, and RET), 2.0 μM (EGFR, erbB2, ErbB3, and ErbB4), 5.0 μM (c-abl), 8.0 μM (Flt-1, Flt-3, Flt-4, Flk, KDR, FGFR-1, and Tek), 10.0 μM (PDGFR-β, protein kinase C-α, and cyclin-dependent kinase 1), and 20.0 μM (c-Src and protein kinase A). The reaction is terminated by the addition of 20 μL 125 mM EDTA. Thirty μL (c-abl, c-Src, insulin-like growth factor-1R, RET-Men2A, and RET-Men2B) or 40 μL (all other kinases) of the reaction mixture is transferred onto Immobilon-polyvinylidene difluoride membrane, presoaked with 0.5% H3PO4 and mounted on a vacuum manifold. Vacuum is then applied and each well rinsed with 200 μL 0.5% H3PO4. Membranes are removed and washed four times with 1.0% H3PO4 and once with ethanol. Dried membranes are counted after mounting in a Packard TopCount 96-well frame and with the addition of 10 μL/well of Microscint. IC50 values (±SE) are calculated by linear regression analysis of the percentage inhibition and are averages of at least three determinations.

Methylene Blue Cell Proliferation Assay.Cells are seeded at 1.5 × 103 cells/well into 96-well microtiter plates and incubated overnight at 37 °C, 5% v/v CO2 and 80% relative humidity. AEE788 dilutions are added on day 1, with the highest concentration being 10 μM. After incubation of the cell plates for an additional 4 (T24) or 6 (BT-474, SK-BR-3, and NCI-H596) days, cells are fixed with 3.3% v/v glutaraldehyde, washed with water, and stained with 0.05% w/v methylene blue. After washing, the dye is eluted with 3% HCl and the absorbance measured at 665 nm with a SpectraMax 340 spectrophotometer. IC50 values are determined by mathematical curve-fitting and are defined as the drug concentration leading to 50% inhibition of net cell mass increase compared with untreated control cultures. (Only for Reference)