NVP-BHG712

製品コードS2202 バッチS220201

印刷

化学情報

Chemical Structure Synonyms N/A Storage
(From the date of receipt)		 3 years -20°C powder
1 years -80°C in solvent
化学式

C26H20F3N7O
分子量 503.48 CAS No. 940310-85-0
Solubility (25°C)* 体外 DMSO 101 mg/mL (200.6 mM)
Ethanol 3 mg/mL (5.95 mM)
Water Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

溶剤液（一定の濃度）を調合する

生物活性

製品説明 NVP-BHG712 is a specific EphB4 inhibitor with ED50 of 25 nM that discriminates between VEGFR and EphB4 inhibition; also shows activity against c-Raf, c-Src and c-Abl with IC50 of 0.395 μM, 1.266 μM and 1.667 μM, respectively.
in vitro NVP-BHG712 treatment also dose dependently leads to the inhibition of RTK autophosphorylation in stable transfected A375 melanoma cells with EC50 of 25 nM and 4.2 μM for EphB4 and VEGFR2, respectively. [1]
in vivo In a growth factor-induced angiogenesis model, NVP-BHG712 (3 mg/kg, p.o) significantly suppresses VEGF stimulated tissue formation and vascularization by inhibiting EphB4 forward signaling. Furthermore, NVP-BHG712 (10 mg/kg/kg, p.o.) potently reverses VEGF enhanced tissue formation and vessel growth. NVP-BHG712 (3 mg/kg, p.o.) shows a long lasting exposure with concentrations around 10 μM in plasma as well as in lung and liver tissue for up to 8 hours, and thus results in a long lasting inhibition of EphB4 kinase activity in mice. [1]
特徴 Discriminates between VEGFR and EphB4.

プロトコル(参考用のみ)

キナーゼアッセイ In vitro kinase assays
All in vitro kinase assays are performed with recombinant purified kinases either purchased from external vendors or produced in house. To estimate kinase activity both, TR-FRET-based LanthaScreenTM and Caliper mobility shift are used. In brief, the LanthaScreenTM assay technology is based on the discrimination between the unphosphorylated substrate and the phosphorylated product by a phospho-specific antibody, binding only to the phosphorylated version of the substrate. Both, antibody and substrate, carry fluorescent labels and the close proximity of the labels in the formed complex allows the measurement of a fluorescence resonance energy transfer (FRET) signal. Reading of the FRET signal in a time-dependent/time-gated manner further improves the assay performance by reducing background fluorescence. For dose–response measurements NVP-BHG712 is pre-diluted in 90% DMSO and 50 nL of compound solutions are dispensed directly into the empty assay plate using a HummingBird nanodispenser. The kinase reactions are started by addition of 4.5 μL ATP solution (4 μM ATP, 20 mM Tris/HCL, 1 mM DTT, 0.03% Tween20, 0.01 mM Na3VO4) and 4.5 μL enzyme/substrate mix (100 nM fluorescein poly-GAT), 0.5% bovine serum albumin, 20 mM Tris/HCL, 1 mM DTT, 0.03% Tween20, 0.01 mM Na3VO4). Further components of the enzyme/substrate mix are the enzymes as well as MgCl2/MnCl2 which are adjusted specifically to the requirements of the individual enzyme. After incubation for 60 minutes at r.t. the kinase reactions are stopped by addition of 4.5 μL stop solution (50 mM EDTA pH 8.0, 0.04% NP-40, 20 mM Tris/HCl pH 7.4) followed by 4.5 μL of detection mix (1.72 μg/mL Tb-PY20 antibody), 1% bovine serum albumin, 20 mM Tris/HCl, 1 mM DTT, 0.03% Tween20, 0.01 mM Na3VO4). After incubation for 45 minutes at r.t. plates are analyzed in a BMG PHERAstar plate reader. In the Caliper mobility shift assays kinase reactions are analyzed by microfluidic capillary electrophoresis. The transfer of phosphate from ATP to a short peptide by a kinase causes a change in the net-charge of the peptide by −2. The charge difference between the unphosphorylated and phosphorylated entities of the peptide can be separated in an electrical field. Using peptides attached with a fluorescent label allows detection and quantification of both forms and hence the calculation of the reaction turnover. For dose–response measurements NVP-BHG712 is pre-diluted in 90% DMSO and 50 nL aliquots of solution are dispensed directly into the empty assay plate using a HummingBird nanodispenser. The kinase reactions are started by addition of 4.5 μL substrate mix consisting of ATP and peptide substrate in assay buffer (50 mM HEPES pH 7.5, 0.02% bovine serum albumin, 1 mM DTT, 0.02% Tween20, 0.01 mM Na34, 10 mM beta-glycerophosphate) and 4.5 μL enzyme solution in assay buffer. The peptide concentration is 2 μM in the assays. Concentrations for the enzyme, as well as for MgCl2 and MnCl2 are adjusted specifically to the requirements of the individual enzyme. ATP concentrations are adjusted to the Km values of the specific enzyme. After incubation for 60 minutes at 30 °C the kinase reactions are stopped by addition of 16 μL stop solution (100 mM HEPES pH 7.5, 5% DMSO, 0.1% coating reagent 10 mM EDTA pH 8.0, 0.015% BRIJ35). Stopped kinase reactions are analyzed in a LC3000 reader.
動物実験 動物モデル VEGF-mediated angiogenesis in vivo is induced in a growth factor implant model in mice.
投薬量 ≤30 mg/kg
投与方法 Administered via p.o.

カスタマーフィードバック

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Selleckの高級品が、幾つかの出版された研究調査結果（以下を含む）で使われた：

EGFR-phosphorylated GDH1 harmonizes with RSK2 to drive CREB activation and tumor metastasis in EGFR-activated lung cancer [ Cell Rep, 2022, 41(11):111827] PubMed: 36516759
Adaptive activation of EFNB2/EPHB4 axis promotes post-metastatic growth of colorectal cancer liver metastases by LDLR-mediated cholesterol uptake [ Oncogene, 2022, 10.1038/s41388-022-02519-z] PubMed: 36376513
Antiviral drug screen identifies DNA-damage response inhibitor as potent blocker of SARS-CoV-2 replication [ Cell Rep, 2021, 35(1):108940] PubMed: 33784499
EFNA4 promotes cell proliferation and tumor metastasis in hepatocellular carcinoma through a PIK3R2/GSK3β/β-catenin positive feedback loop [ Mol Ther Nucleic Acids, 2021, 25:328-341] PubMed: 34484860
Targeting the EphB4 receptor tyrosine kinase sensitizes HER2-positive breast cancer cells to Lapatinib. [ Cancer Lett, 2020, 475:53-64] PubMed: 32006616
Identification of SYK Inhibitor, R406 as a Novel Senolytic Agent [ Aging (Albany NY), 2020, 7;12(9):8221-8240] PubMed: 32379705
EPHB4 inhibition activates ER stress to promote immunogenic cell death of prostate cancer cells. [ Cell Death Dis, 2019, 10(11):801] PubMed: 31641103
Deep Learning Enhancing Kinome-Wide Polypharmacology Profiling: Model Construction and Experiment Validation. [ J Med Chem, 2019, 10.1021/acs.jmedchem.9b00855] PubMed: 31364850
Mitochondrial metabolic reprograming via BRAF inhibition ameliorates senescence. [ Exp Gerontol, 2019, 126:110691] PubMed: 31421186
Comparative analysis of the phototoxicity induced by BRAF inhibitors and alleviation through antioxidants. [ Photodermatol Photoimmunol Photomed, 2019, 10.1111/phpp.12520] PubMed: 31618797

長期の保管のために-20°Cの下で製品を保ってください。

人間や獣医の診断であるか治療的な使用のためにでない。

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