| 名稱 | NVP-BVU972 |
| 描述 | NVP-BVU972 is a selective and potent Met inhibitor with IC50 of 14 nM. |
| 細(xì)胞實(shí)驗(yàn) | BaF3 cells containing TPR-MET or various mutants thereof are grown in RPMI 1640 medium containing 10% fetal calf serum. For maintenance of parental BaF3 cells the medium is additionally supplemented with 10 ng/mL interleukin-3 (IL-3). For proliferation assays, BaF3 cells are seeded on 96-well-plates in triplicates at 104 cells per well and incubated with various concentrations of NVP-BVU972 for 72 hours followed by quantification of viable cells using a resazurin sodium salt dye reduction readout. IC50 values are determined with the XLFit Excel Add-In using a 4-parameter dose response model.(Only for Reference) |
| 激酶實(shí)驗(yàn) | TR-FRET biochemical assay with MET wild type and mutants: Enzyme activity is measured in a time resolved fluorescence resonance energy transfer (TR-FRET) assay, detecting tyrosine phosphorylation with a Eu-labelled anti-phospho-tyrosine antibody (fluorescence donor) and Allophycocyanin conjugated to Streptavidin (fluorescence acceptor) which binds to a biotin on the substrate peptide. For each variant, Km concentrations for ATP are determined in the absence of NVP-BVU972, and the ATP concentration in the kinase reaction is set to Km (4 μM for MET wt, 1 μM for MET Y1230H and MET F1200I). NVP-BVU972 is dissolved and diluted in DMSO and assayed in quadruplicate. Kinase reactions are carried out in 50 mM Tris-HCl pH 7.5, 8 mM MgCl2, 4 mM MnCl2, 0.05 % Tween 20, 0.05% bovine serum albumin, 0.1 mM EDTA, 1 mM DTT, 0.1 mM Na3VO4, in white 1536 well plates at room temperature. NVP-BVU972 and enzyme are incubated in a volume of 2 μL for 20 min, followed by the addition of 1 μL ATP and 1 μL biotinylated peptide substrate (PTK1) to final concentrations of Km and 1 μM, respectively. Enzyme concentrations in the reactions are 5 nM for MET wt, and 4 nM for the F1200I and Y1230H variants. After 90 min, reactions are stopped by addition of 1 μL stop/detection solution to reach final concentrations of 10 mM EDTA, 3.5 nM Eu-labelled antiphospho-tyrosine antibody PY20, and 10 nM Streptavidin Allophycocyanin. Time resolved fluorescence resonance energy transfer is measured in an Envision plate reader (excitation 320 nm, emission 615 nm and 665 nm). |
| 體內(nèi)活性 | NVP-BVU972有效抑制MET基因擴(kuò)散的細(xì)胞系GTL-16,MKN-45和EBC-1的生長,IC50分別為66,82和32 nM。NVP-BVU972有效抑制MET激酶,而對其他激酶（包括最密切相關(guān)的激酶）RON抑制作用很低,IC50>1000 nM。NVP-BVU972抑制GTL-16細(xì)胞中組成型MET磷酸化,抑制A549細(xì)胞中HGF刺激的MET磷酸化,IC50分別為7.3和 22 nM。NVP-BVU972作用于表達(dá)野生型TPR-MET的BaF3細(xì)胞,劑量依賴性降低TPR-MET磷酸化。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 45 mg/mL (132.21 mM), Sonication is recommended.
Ethanol : 63 mg/mL (185.1 mM)
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | inhibit | NVP-BVU 972 | NVP-BVU972 | BaF3 TPR-MET | phosphorylation | cancer | BaF3 | c-Met/HGFR | anti-proliferation | MET resistance mutations | Inhibitor | NVP-BVU-972 | NVP BVU972 | NVPBVU972 |
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| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 抗結(jié)直腸癌化合物庫 | 激酶抑制劑庫 | 高選擇性抑制劑庫 | 抗肝癌化合物庫 | 抗肺癌化合物庫 | 膜蛋白靶向化合物庫 | 酪氨酸激酶分子庫 |