AMG458

• 产品号: S2747
• CAS号: 913376-83-7
• 分子式: C30H29N5O5
• 分子量: 539.58176
• 保 存: -20°C

产品别名

• 标题: AMG458
• IUPAC标准名: 1-(2-hydroxy-2-methylpropyl)-N-{5-[(7-methoxyquinolin-4-yl)oxy]pyridin-2-yl}-5-methyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazole-4-carboxamide
• IUPAC传统名: 1-(2-hydroxy-2-methylpropyl)-N-{5-[(7-methoxyquinolin-4-yl)oxy]pyridin-2-yl}-5-methyl-3-oxo-2-phenylpyrazole-4-carboxamide

产品登记号

• CAS号: 913376-83-7

产品性质

• 作用靶点: c-Met
• 成盐信息: Free Base
• 保存条件: -20°C

产品详细信息

详细说明 (English)

Research Area

• Description: Cancer

Biological Activity

• Description: AMG 458 is a potent c-Met inhibitor of human c-Met and mouse c-Met with K_i of 1.2 nM or 2.0 nM.
• Targets: Human c-Met; Mouse c-Met
• IC50: 1.2 nM (K_i); 2.0 nM （K_i) ^[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 (Cl_int: <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: AMG 458 is 100% bioavailable across species and the intrinsic half-life increased with higher mammals.

Protocol

• Protocol: Kinase Assay ^[1]
• 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 K_m 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.
• 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 × 10^4 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.
• Protocol: Animal Study ^[1]
• Animal Models: NIH3T3/TPR-Met and U-87 MG xenograft models are established in female CD-1 nu/nu mice aged 6-8 weeks.
• Formulation: i.v. dose: 1 mg/kg (20% Captisol with pH adjusted to 3.5 using methanesulfonic acid). p.o. dose: 10 mg/kg (2% HPMC and 1% Tween-80 with pH adjusted to 2.2 using HCl). Both are solution formulations with the same drug concentration of 1 mg/mL.
• Doses: 10, 30 or 100 mg/kg
• Administration: Administered via i.v. or p.o.

References

• References: [1] Liu L, et al. J Med Chem, 2008, 51(13), 3688-3691.
• References: [2] Teffera Y, et al. Chem Res Toxicol, 2008, 21(11), 2216-2222.
• References: [3] Torossian A, et al. SS M-Biology, 2011.

参考文献

• Liu L, et al. J Med Chem, 2008, 51(13), 3688-3691.
• Teffera Y, et al. Chem Res Toxicol, 2008, 21(11), 2216-2222.
• Torossian A, et al. SS M-Biology, 2011.