| 名稱 | PF-562271 |
| 描述 | PF-562271 is an effective ATP-competitive, reversible inhibitor of FAK(IC50=1.5 nM) and Pyk2 kinase(IC50=13 nM). |
| 細(xì)胞實(shí)驗(yàn) | PF-562271 (Haoyuan Chemexpress Co., Ltd.) is dissolved in DMSO and stored, and then diluted with appropriate media before use[2]. Ewing sarcoma cells are plated in 10-cm dishes, allowed to adhere for 24 hours, and then treated with PF-562271, PD0325901, or Dasatinib. ATP content is measured as a surrogate for cell number using the CellTiter-Glo Luminescent Cell Viability Assay. Luminescence readings are obtained using the FLUOstar Omega microplate reader. For experiments with small-molecule treatment, 1.25×103 Ewing sarcoma cells are seeded in each well and treated with a range of concentrations. IC50 values are calculated from ATP measurements obtained after 3 days of treatment using log-transformed, normalized data in GraphPad Prism 5.0. Cell lines are also treated with compound in 6-cm dishes, trypsinized, and counted by light microscopy using trypan blue exclusion. For experiments using shRNA-transduced cells, 1.25×103 cells are seeded per well into 384-well plates on day 3 posttransduction. ATP content is measured on days 3, 6, and 8 posttransduction[2]. |
| 激酶實(shí)驗(yàn) | The purified-activated FAK kinase domain (amino acid 410-689) is reacted with 50 μM ATP and 10 μg per well of a random peptide polymer of Glu and Tyr, p(Glu/Tyr), in kinase buffer (50 mM HEPES pH 7.5, 125 mM NaCl, and 48 mM MgCl2) for 15 min. Phosphorylation of p(Glu/Tyr) is challenged with serially diluted compound at 1/2-Log concentrations starting at a top concentration of 1 μM. Each concentration is tested in triplicate. Phosphorylation of p(Glu/Tyr) is detected with a general antiphospho-tyrosine (PY20) antibody followed by horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG antibody. HRP substrate is added, and absorbance readings at 450 nm are obtained after addition of stop solution (2 M H2SO4). IC50 values are determined using the Hill-Slope Model[1]. |
| 體外活性 | 在脛骨植入MDA-MB-231細(xì)胞的大鼠中,口服 PF-562271（5 mg/kg）,引起骨鈣素和松質(zhì)骨增加,從而使腫瘤細(xì)胞生長減緩.在攜帶H125肺部異種移植腫瘤、異種移植PC3M-luc-C6的小鼠模型中,口服 PF-562271 （25 mg/kg）抑制腫瘤細(xì)胞生長,引起細(xì)胞凋亡.在U87 mg的小鼠中,PF-562271（口服< 33 mg/kg）能夠以時(shí)間和劑量依賴的方式抑制腫瘤中FAK磷酸化.在BxPc3異種移植小鼠、PC3-M異種移植小鼠中口服 PF-562271（50 mg/kg）,能夠抑制腫瘤生長. |
| 體內(nèi)活性 | PF-562271能夠結(jié)合在ATP與FAK結(jié)合的部位,并在激酶鉸鏈區(qū)形成抑制劑與主鏈原子之間的氫鍵。在雞胚絨毛尿囊膜中,PF-562271（1 nM ）阻斷bFGF刺激的血管生成。在PC3-M細(xì)胞中,PF-562271（3.3 μM）能夠使細(xì)胞停止在G1期。對(duì)于A431 細(xì)胞,PF-562271（250 nM ）能夠抑制細(xì)胞侵入膠原蛋白。 |
| 存儲(chǔ)條件 | store at low temperature | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 93 mg/mL (183.3 mM)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | Cancer | PTK2 | Ewing | Antitumor | Pyk2 | PTK2 protein tyrosine kinase 2 | Sarcoma | inhibit | PF-562271 | Breast | FAK | Focal adhesion kinase | VS 6062 | VS6062 | Inhibitor | Proline-rich tyrosine kinase 2 | VS-6062 |
| 相關(guān)產(chǎn)品 | Sodium Oxamate | Ribociclib | Palbociclib monohydrochloride | Abemaciclib methanesulfonate | CASIN | Olomoucine | Palbociclib | GW 441756 | Dinaciclib | Defactinib | Ro-3306 | Abemaciclib |
| 相關(guān)庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 酪氨酸激酶分子庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |