NVP-ADW742

Injection i.p. or oral gavage

MM-1S, MM-1R, RPMI-8226/S, OPM-1, OCI-My5, SKMM2, , KMS-12-BM, XG-1, L363, S6B45 cells and et al.

Dissolved in DMSO, final concentrations ca. 10 uM

10 mg/kg twice daily

0.17 uM [1], 0.17 uM [1], 0.17 uM [1], 0.17 uM [1], 0.17 uM [1], 0.17 uM [1]

Administration of NVP-ADW742 at 10 mg/kg twice daily significantly inhibits tumor growth, prolongs survival, and enhances the antitumor effect of cytotoxic chemotherapy melphalan in the mice model of diffuse MM. [1]

Cellular kinase activity assay, The IC50 value for the effect of NVP-ADW742 on the autophosphorylation of IGF-1R is determined at the cellular level in the presence of increasing concentrations of NVP-ADW742, using 96-well “Capture ELISAs” assays. Briefly, NWT-21 cells are seeded into 96-well tissue culture plates in complete growth medium and grown to 70-80% confluency, and are then starved for 24 hours in 0.5% FCS medium. Subsequently, cells are incubated for 90 minutes in the presence of NVP-ADW742 followed by the stimulation with of IGF-I (10 ng/mL) for 10 minutes at 37 C. Subsequently, the cells are washed twice with ice-cold PBS and lysed at 4 C with 50 uL/well RIPA-buffer (50 mM Tris-HCl, pH 7.2, 120 mM NaCl, 1 mM EDTA, 6 mM EGTA, 1% NP-40, 20 mM NaF, 1 mM benzamidine, 15 mM sodium pyrophosphate, 1 mM PMSF and 0.5 mM Na3VO4). The lysates from each experiment are then transferred to black ELISA plates precoated with capture antibodies specific for IGF-1R. After capture by the antibodies, lysates aixed with 40 uL of an alkaline phosphatase (AP) labelled anti-phosphotyrosine Ab (PY20(AP) diluted to 0.2 ug/mL in RIPA buffer, and incubated overnight at 4 C. After washing (PBST) and incubation for 45 minutes at RT with the luminescent AP-substrate CDPStar RTU with Emerald II (90 uL/well), luminescence is measured using a Packard Top Count Scintillation Counter.

453, 58

DMSO 10 mg/mL, Water <1 mg/mL, Ethanol 3 mg/mL