AEE788 (NVP-AEE788)

ERBB2, EGFR

440, 58

NCI-H596, DU145, A431, B16 and oncogenic NeuT-transfected HC11 cells in female BALB/c nu/nu (nude) mice

A Phase I/II study in adult patients with recurrent or relapsing glioblastoma multiforme has been completed.

50 mg/kg

2 nM, 2 nM, 2 nM, 2 nM, 2 nM, 2 nM

AEE788 produces a dose-dependent inhibition of tumor growth in NCI-H596 or DU145 xenograft models, with only minor body weight changes. AEE788 induces tumor regression by 57% at 50 mg/kg in the NeuT/erbB2 GeMag model. AEE788 potently inhibits EGF-induced EGFR phosphorylation in A431 tumors and erbB2 phosphorylation in GeMag tumors. AEE788 dose-dependently inhibited angiogenesis induced by VEGF and does not inhibit bFGF-induced angiogenesis. [1] AEE788 suppresses the growth of tumor volume by 54% in Colo16 xenografts at 50 mg/kg, which dues to the inhibition of phosphorylation of EGFR, VEGFR, Akt, and MAPK. [2] AEE788 (50 mg/kg) also inhibits growth of tumors in the cecum and peritoneum (>50%) and reduces the incidence of lymph node metastasis to 70% in HT29 cells implanted in the cecum of nude mice, without loss of body weight and gross evidence of neovascularization. AEE788 significantly lowers the expression levels of pEGFR and pVEGFR in HT29 cecal tumors and does not alter those of EGF, VEGF, EGFR, or VEGFR. Combined with CPT-11, AEE788 has significantly smaller tumors and complete inhibition of lymph node metastasis. [3] AEE788 inhibits the growth of Daoy, DaoyPt, and DaoyHER2 xenografts by 51%, 45%, and 72%, respectively. [4] AEE788 could promote LBH589-mediated generation of reactive oxygen species in K562 tumor cells, which in turn increase apoptosis. [5]

Protein Kinase Assays, The in vitro kinase assays are performed in 96-well plates (30 uL) at ambient temperature for 15–45 min using the recombinant glutathione S-transferase-fused kinase domains (4-100 ng, depending on specific activity). [gamma33P]ATP is used as phosphate donor and polyGluTyr-(4:1) peptide as acceptor. With the exception of protein kinase C-alpha, cyclin-dependent kinase 1/cycB and protein kinase A are protamine sulfate (200 ug/mL), histone H1 (100 ug/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 uM (c-Kit, c-Met, c-Fms, c-Raf-1, and RET), 2.0 uM (EGFR, erbB2, ErbB3, and ErbB4), 5.0 uM (c-abl), 8.0 uM (Flt-1, Flt-3, Flt-4, Flk, KDR, FGFR-1, and Tek), 10.0 uM (PDGF receptor-beta, protein kinase C-alpha, and cyclin-dependent kinase 1), and 20.0 uM (c-Src and protein kinase A). The reaction is terminated by the addition of 20 u125 mM EDTA. Thirty uL (c-abl, c-Src, insulin-like growth factor-1R, RET-Men2A, and RET-Men2B) or 40 uL (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 uL 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 uL/well of Microscint. IC50 values (+/-SE) are calculated by linear regression analysis of the percentage inhibition and are averages of at least three determinations.

DMSO 88 mg/mL, Water <1 mg/mL, Ethanol <1 mg/mL

(R)-6-(4-((4-ethylpiperazin-1-yl)methyl)phenyl)-N-(1-phenylethyl)-7H-pyrrolo[2, 3-d]pyrimidin-4-amine