Antibodies
AMG-458
- 貨號 Cat. No.
- S2747
- 規格 Size
- 5~200mg
- 品牌 Brand
- Selleckchem
Description
AMG-458
Catalog No.S2747
AMG 458 is a potent c-Met inhibitor with Ki of 1.2 nM, ~350-fold selectivity for c-Met than VEGFR2 in cells.
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Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw
Biological Activity
| Description | AMG 458 is a potent c-Met inhibitor with Ki of 1.2 nM, ~350-fold selectivity for c-Met than VEGFR2 in cells. | |||||
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| Targets | Human c-Met | Mouse c-Met | ||||
| IC50 | 1.2 nM (Ki) | 2.0 nM （Ki) [1] | ||||
| In vitro | AMG 458 also inhibits HGF-mediated c-Met phosphorylation in PC3 and CT26 cells with IC50 of 60 and 120 nM.[1] AMG 458 is observed to bind covalently to liver microsomal proteins from rats and humans in the absence of NADPH. AMG 458 is believed to react with thiol groups in proteins, producing a methoxy quinoline thioether conjugate. [2] A recent study shows that the constitutive phosphorylation of c-Met in H441 is abrogated by AMG 458. The basal and HGF-induced phosphorylation of c-Met in A549 is attenuated by AMG 458. The combination of radiation therapy and AMG 458 treatment is found to synergistically increase apoptosis in the H441 cell line by reduction of p-Akt and p-Erk levels, but not in A549. [3] | |||||
| In vivo | AMG 458 is metabolically stable in the liver microsomes of mouse, rat, dog, monkey, and human with low intrinsic clearances (Clint: <5, 62, 8, 8, 18 (μL/min)/mg, respectively). When administered orally, AMG 458 achieves remarkably high bioavailability in all species tested. Oral dosing of AMG 458 inhibits HGF-mediated c-Met phosphorylation with an approximate ED90 of 30 mg/kg and an associated plasma exposure of approximately 15 μM at 6 hours. AMG 458 significantly inhibits tumor growth in the NIH3T3/TPR-Met and U-87 MG xenograft models at 30 and 100 mg/kg q.d. and 30 mg/kg b.i.d.with no adverse effect on body weight. [1]High concentrations of AMG 458 in some organs may produce toxicity via oxidative stress. [2] | |||||
| Features | Completely bioavailable across species and the intrinsic half-life is increased in higher mammals. | |||||
Protocol(Only for Reference)
Kinase Assay:
| In vitro kinase assay | Optimal enzyme, ATP, and substrate (gastrin peptide) concentrations are established for each enzyme using homogeneous time-resolved fluorescence (HTRF) assays. AMG 458 is tested in a 10-point dose-response curve for each enzyme using an ATP concentration of two-thirds Km for each. Most assays consisted of enzyme mixed with kinase reaction buffer [60 mM HEPES, pH 7.4, 50 mM NaCl, 20 mM MgCl2, and 5 mM MnCl2]. A final concentration of 1 mM DTT, 0.2 mM Na3VO4, and 20 μg/mL BSA is added before each assay. For all assays, 5.75 mg/mL streptavidin-allophycocyanin and 0.1125 nM Eu-PT66 are added immediately before the HTRF reaction. Plates are incubated for 30 minutes at room temperature and the fluorescence ratios are read on a Rubystar instrument. |
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| Cell-based autophosphorylation assay of c-Met | PC3 and CT23 cells are grown as monolayers using standard cell culture conditions. IC50 measurement of AMG 458 activities on HGF-mediated c-Met autophosphorylation is determined in serum-starved PC-3 (human) or CT26 (mouse) cells using a quantitative electrochemiluminescent immunoassay. PC3 and CT26 cells are plated at a density of 2 × 104 cells/well in 96 well-plates. 24 hours after plating, cells are starved in media containing 0.1% BSA for 18 to 20 hours. Cells are then treated with a 10-point serial dilution of AMG 458 for one hour at 37 °C followed by stimulation with optimal concentrations of recombinant human HGF for 10 minutes at 37 °C. Cells are washed once with PBS and lysed (1% Triton X-100, 50 mM Tris pH 8.0, 100 mM NaCl, 300 μM Na3VO4 and protease inhibitors). Cell lysates are incubated with a biotin-labeled goat–anti -c-Met antibody (BAF358- for human c-Met -, BAF527- for mouse c-Met -) for capture followed by a mouse anti-phosphotyrosine antibody 4G10 and a BV-tagTM labeled anti-mouse IgG as the detection antibody. Levels of c-Met phophorylation are then measured on a BioVeris M-Series instrument. The IC50 values are calculated using Xlfit4-parameter equation. |
Animal Study:
| Animal Models | NIH3T3/TPR-Met and U-87 MG xenograft models are established in female CD-1 nu/nu mice aged 6-8 weeks. |
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| Dosages | 10, 30 or 100 mg/kg |
| Administration | Administered via i.v. or p.o. |
| Solubility | 15% Captisol/citrate vehicle, 30 mg/mL |
References
Chemical Information
| Molecular Weight (MW) | 539.58 |
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| Formula | C30H29N5O5 |
| CAS No. | 913376-83-7 |
| Storage | 3 years -20℃Powder |
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| 6 months-80℃in DMSO | |
| Syonnyms | N/A |
| Solubility (25°C) * | In vitro | DMSO | 21 mg/mL (38 mM) |
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| Water | <1 mg/mL (<1 mM) | ||
| Ethanol | <1 mg/mL (<1 mM) | ||
| In vivo | 15% Captisol/citrate vehicle, | 30 mg/mL | |
| * <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. | |||
| Chemical Name | 1-(2-hydroxy-2-methylpropyl)-N-(5-(7-methoxyquinolin-4-yloxy)pyridin-2-yl)-5-methyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazole-4-carboxamide |
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Preparing Stock Solutions
| Stock Solution (1ml DMSO) | 1mM | 10mM | 20mM | 30mM |
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| Mass(mg) | 0.53958 | 5.3958 | 10.7916 | 16.1874 |
