| 名稱 | Pelitinib |
| 描述 | Pelitinib (EKB-569) (EKB-569) is an effective irreversible EGFR inhibitor (IC50: 38.5 nM). |
| 細(xì)胞實(shí)驗(yàn) | Cells are seeded in 96-well dishes, and after 2 hours, Pelitinib is added and incubated for 5 days. After incubation, the medium is removed from each well and fresh medium (150 μL) + 1 mg/mL MTT solution (50 μL) is added. After incubation for 2 hours at 37 °C, the medium is replaced with 150 μL DMSO, and absorbance at 540 nm in each well is determined. The IC50 is calculated by linear regression of the data. (Only for Reference) |
| 激酶實(shí)驗(yàn) | Autophosphorylation of EGFR in cells: For experiments using cells in culture, A431 cells are treated with various concentrations of Pelitinib for 2.75 hours before co-incubation with 100 ng/mL EGF for 0.25 hour. Cells are washed twice with cold phosphate-buffered saline (PBS) before adding to lysis buffer (10 mM Tris, pH 7.5, 5 mM ethylenediamine tetra-acetic acid (EDTA), 150 mM NaCl, 1% Triton X-100, 1% Sodium deoxycholate, 0.1 % SDS, 1 mM PMSF, 10 mg/mL pepstatin A, 10 mg/mL leupeptin, 20 KIU/mL aprotinin, 2 mM sodium orthovanadate, and 100 mM sodium fluoride) for 20 minutes on ice, before immunoprecipitation and SDS-PAGE-immunoblotting. For immunoprecipitation, cultured cells are placed in cold lysis buffer and immediately homogenized on ice with a polytron with several pulses. The homogenate is first centrifuged at 2500 rpm (20 minutes, 4 °C) and then again at 14,000 rpm in a microcentrifuge (10 minutes, 4 °C). Supernatants (1000 μg protein) are incubated for 2 hours at 4 °C with 15 mL of EGFR polyclonal antibody. After 2 hours, 50 μL of protein G plus/protein A agarose beads is added and incubated with constant rotation for 2 hours at 4 °C. After washing with lysis buffer, beads are boiled for 2 minutes in Laemmli sample buffer. Proteins are then resolved by SDS-PAGE, transferred to immobilon membrane and probed overnight with an anti-phosphotyrosine antibody conjugated with horseradish peroxidase (HRP). Membranes are developed using the ECL reagent. Total EGFR protein is determined by stripping membranes and re-probing with receptor-specific antibodies. Quantitation of bands is done by densitometry, using ImageQuant software with a Molecular Dynamics laser transmittance scanner. |
| 體外活性 | 在達(dá)EGFR的A431異種移植模型中,Pelitinib（10 mg/kg,p.o.）可有效抑制EGFR磷酸化,1 h內(nèi)抑制90%,24 h后抑制率>50%.小鼠氣道上皮細(xì)胞重構(gòu)模型,可被病毒感染誘導(dǎo),具有延遲但永久性轉(zhuǎn)變?yōu)楸瓲罴?xì)胞化生的特點(diǎn),Pelitinib（20 mg/kg/day）可完全阻斷纖毛細(xì)胞的增加和Clara細(xì)胞的減少,從而從3個方面校正上皮細(xì)胞重構(gòu),并明顯抑制杯狀細(xì)胞的發(fā)生.Pelitinib（20 mg/kg/day）抑制APCMin/+小鼠體內(nèi)87%的腫瘤發(fā)生,與2倍劑量EKI-785（40 mg/kg/day）的作用接近,與體內(nèi)活性較高一致. Pelitinib對體內(nèi)氣道上皮細(xì)胞中EGFR信號有選擇性抑制效果. |
| 體內(nèi)活性 | Pelitinib（75-500 nM）可特異性抑制AKT和ERK1/2的活化,而對NF-κB通路無影響。在正常人角質(zhì)細(xì)胞中,Pelitinib也可有效抑制TGF-α介導(dǎo)的EGFR活化（IC50：56 nM）,抑制STAT3和ERK1/2的活化（IC50：60 nM和62 nM）。在A431和正常人角質(zhì)細(xì)胞中,Pelitinib對EGF誘導(dǎo)的EGFR磷酸化有抑制作用（IC50：20-80 nM）,還抑制STAT3的磷酸化（IC50：30-70 nM）。 Pelitinib對正常人角質(zhì)細(xì)胞（IC50：61 nM）,以及A431（IC50：125 nM）和MDA-468（IC50：260 nM）腫瘤細(xì)胞的增殖有明顯抑制作用,但對MCF-7細(xì)胞活性較低（IC50：3.6 μM）。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : < 1 mg/mL (insoluble or slightly soluble)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 4.68 mg/mL (10 mM), Sonication is recommended.
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| 關(guān)鍵字 | inhibit | Src | EKB569 | HER1 | Pelitinib | Epidermal growth factor receptor | Inhibitor | WAY-EKB-569 | ErbB-1 | EGFR | WAY-EKB569 | EKB 569 |
| 相關(guān)產(chǎn)品 | Lidocaine hydrochloride | Lapatinib | Neratinib | Lidocaine | Gefitinib | Erlotinib | Osimertinib | Erlotinib hydrochloride | Dabrafenib | Genistein | Regorafenib | Sorafenib |
| 相關(guān)庫 | 抗癌活性化合物庫 | 經(jīng)典已知活性庫 | 激酶抑制劑庫 | 膜蛋白靶向化合物庫 | 藥物功能重定位化合物庫 | 酪氨酸激酶分子庫 | 疼痛相關(guān)化合物庫 |