| 名稱 | CP-673451 |
| 描述 | CP-673451 is a specific inhibitor of PDGFRα/β (IC50: 10/1 nM) with antiangiogenic and antitumor activity and the selectivity is higher 450-fold than other angiogenic receptors. |
| 細(xì)胞實(shí)驗(yàn) | PAE cells stably expressing full-length PDGFR and VEGFR have been generated. For cell-based selectivity assays, PAE cells are transfected with fulllength human PDGFR-a, PDGFR-h, or VEGFR-2. Cells are seeded at 4×105 cells/mL in 50 μL growth medium (Ham's F-12 media supplemented with 10% fetal bovine serum, 50,000 units each penicillin and streptomycin, and 500 μg/mL gentamicin) per well in 96-well plates. After 6 to 8 hours, the growth medium is replaced with 50 μL serum-depleted medium (as above, but with 0.1% fetal bovine serum) and cells are incubated overnight. Immediately before compound addition, the medium was replaced with 95 μL serum-depleted medium. Compounds are diluted in 100% DMSO, added to the cells at a final DMSO concentration of 0.25% v/v, and incubated at 37°C for 10 minutes. Cells are stimulated with the appropriate ligand and incubated as above for an additional 8 minutes. The medium is removed and the cells washed once with PBS, then lysed with 50 μL HNTG buffer [20 mmol/L HEPES (pH 7.5), 150 mmol/L NaCl, 2% Triton X-100, 10% glycerol, 5 μmol/L EDTA, 2 mmol/L NaVO4, and 1 EDTA-free complete protease inhibitor tablet per 25 mL] for 5 minutes at room temperature. Lysates are then diluted with 50 μL HG buffer [20 mmol/L HEPES (pH 7.5), 10% glycerol]. The diluted cell lysates are mixed thoroughly, 50 μL of supernatant are transferred to the ELISA capture plate, and incubated at room temperature for 2 hours with agitation. ELISA capture plates are prepared by coating 96-well ReactiBind goat-antirabbit plates with 100 μL/well of 5 μg/mL rabbit anti-human PDGFR-h, anti-PDGFR-a, or anti-VEGFR-2 antibody for 60 to 90 minutes. At the end of the 2-hour incubation the plates are washed (PBS, 0.1% Tween 20) before incubation with anti-phosphotyrosine-horseradish peroxidase antibody (diluted in PBS, 0.05% Tween 20) for 30 minutes at room temperature. The plates are washed again, then incubated with tetramethylbenzidine and evaluated as described above.IC50 values are calculated as percent inhibition of control.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Kinase inhibition assay: A glutathione S-transferase-tagged kinase domain construct of the intracellular portion of the PDGFR-β (amino acids 693-1401, accession no. J03278) is expressed in Sf-9 cells (baculovirus expression system). Enzyme kinetics are determined by incubating the enzyme with increasing concentrations of ATP in phosphorylation buffer [50 mmol/L HEPES (pH 7.3), 125 mmol/L NaCl, 24 mmol/L MgCl2 in Nunc Immuno MaxiSorp 96-well plates previously coated with 100 μL of 100 μg/mL poly-Glu-Tyr (4:1 ratio) diluted in PBS. After 10 minutes, the plates are washed (PBS, 0.1% Tween 20), incubated with anti-phosphotyrosine-horseradish peroxidase antibody, and diluted in PBS, 0.05% Tween 20, 3% BSA for 30 minutes at room temperature. The plates are washed as above and incubated with 3,3',5,5'-tetramethylbenzidine. The reaction is stopped by adding an equal volume of 0.09 NaH2SO4. The phosphotyrosine-dependent signal is then quantitated on a plate reader at 450 nm. For routine enzyme assays, the enzyme is incubated with 10 μM ( final) ATP in the presence of compound diluted in DMSO (1.6% v/v DMSO assay final) for 30 minutes at room temperature in plates, as above, previously coated with 100 μL of 6.25 μg/mL poly-Glu-Tyr. The remainder of the assay is carried out as above, and IC50 values are calculated as percent inhibition of control. |
| 體外活性 | CP 673451是一種選擇性的PDGFRα/β抑制劑，其IC50分別為10 nM/1 nM，對(duì)其他血管生成受體具有超過450倍的選擇性。在膠質(zhì)母細(xì)胞瘤腫瘤中，CP-673451（33 mg/kg）能在4小時(shí)內(nèi)使PDGFR-β受體的抑制率超過50%，相應(yīng)的在血漿Cmax時(shí)EC50為120 ng/mL。在海綿血管生成模型中，CP-673451以3 mg/kg劑量（每天一次，經(jīng)口給藥，對(duì)應(yīng)Cmax時(shí)5.5 ng/mL）抑制了70%的PDGF-BB刺激的血管生成。CP-673451通過減少GSK-3α和GSK-3β的磷酸化來降低細(xì)胞增殖率。在RD和RUCH2細(xì)胞培養(yǎng)中，CP-673451損害了成瘤球形成能力和細(xì)胞分化，導(dǎo)致細(xì)胞衰老增加。 |
| 體內(nèi)活性 | CP 673451 (每日一次口服)在多種植入于無胸腺小鼠皮下的人類腫瘤異種移植物中抑制腫瘤生長(ED50 < 33 mg/kg)，包括H460人類肺癌、Colo205和LS174T人類結(jié)腸癌以及U87 mg人類膠質(zhì)母細(xì)胞瘤。[1] 在攜帶RUCH2異種移植物的小鼠中，CP 673451減少了腫瘤生長及間質(zhì)細(xì)胞浸潤。[2] |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : 41.8 mg/mL (100 mM)
DMSO : 18.33 mg/mL (43.91 mM)
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| 關(guān)鍵字 | Inhibitor | CP673451 | PDGFR | CP-673451 | inhibit | CP 673451 | Platelet-derived growth factor receptor |
| 相關(guān)產(chǎn)品 | Imatinib | Gilteritinib | Ribociclib | Axitinib | Sorafenib tosylate | Lenvatinib mesylate | Regorafenib | Pazopanib | Nintedanib | Sorafenib | Pexidartinib | Regorafenib monohydrate |
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