| 名稱 | Nazartinib |
| 描述 | Nazartinib (EGF816) (EGF816, NVS-816) is a covalent, irreversible, mutant-selective EGFR inhibitor that has nanomolar inhibitory potency against activating mt (L858R, ex19del) and T790M mt, with up to 60-fold selectivity over wild type (wt) EGFR in vitro. |
| 細(xì)胞實(shí)驗(yàn) | H1975, H3255, HCC827, A431, and HaCaT cells are maintained in RPMI media supplemented with antibiotics and 10% FBS, maintained in a 37°C, 5% CO2 humidified incubator. After an overnight incubation in 384-well plates, serial diluted compounds are transferred to cells and incubated for 3 hours. HaCaT cells are stimulated with 10 ng/mL EGF (50 ng/mL EGF for A431) for 5 minutes. Cells are lysed in 1% Triton X-100 buffer containing protease and phosphatase inhibitors. Lysates are analyzed by sandwich ELISA utilizing goat anti-EGFR capture antibody, anti-phospho-EGFR(Y1173), and anti-rabbit HRP. Signal is measured by chemiluminescent detection.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Recombinant kinase domain of EGFR L858R and T790M-L858R mutants are incubated with EGF816 to confirm covalent modification of EGFR and site of adduction. Recombinant enzyme is incubated at room temperature with a 20-fold molar excess of compound in 40 mM Tris, pH 8, 500 mM NaCl, 1% glycerol, 5 mM TCEP for 1 h. The reaction is quenched by addition of dithiothreitol (DTT, 80-fold excess to compound) and transfer to ice. A third of the reaction (10 μL) is processed for intact MS by adding an equal volume of 6 M Guan HCl, 100 mM Tris, pH 8, 20 mM DTT, 10 mM TCEP and incubating at room temperature for 15 min. Intact MS analysis is performed on an Agilent 6520 QToF mass spectrometer equipped with a dual spray ion source (IS of 4500 V, fragmentor of 250 V, fas temp of 350°C, and skimmer of 75 V). The samples are injected onto a PLRP-S column (2.1 mm × 50 mm), heated to 60°C, and desalted for 2 min at 500 μL/min and 3% B prior to elution with a fast gradient of 3-50% B in 3 min (B, 0.1% formic acid). The data are analyzed in MassHunter for automatic peak selection, integration, and spectral deconvolution with a mass range of 15?000-75?000 Da. |
| 體外活性 | Nazartinib是一種新型共價(jià)的、突變選擇性EGFR抑制劑，對(duì)致癌基因(L858R和ex19del)和T790M耐藥性突變具有幾乎等效的活性，并對(duì)WT EGFR具有良好的選擇性。Nazartinib在體外對(duì)最常見的EGFR突變L858R、Ex19del和T790M表現(xiàn)出強(qiáng)大的抑制作用。通過使用攜帶L858R、Ex19del和L858R/T790M突變的三種特征明確的細(xì)胞系H3255、HCC827和H1975，評(píng)估了Nazartinib對(duì)EGFR突變體的細(xì)胞活性。與細(xì)胞孵育3小時(shí)后，Nazartinib在H3255、HCC827和H1975細(xì)胞中顯示出對(duì)pEGFR水平的強(qiáng)效抑制，其EC50值分別為5、1和3nmol/L。基于細(xì)胞的測(cè)定顯示Nazartinib對(duì)突變型EGFR比對(duì)WT EGFR具有選擇性。 |
| 體內(nèi)活性 | Nazartinib在小鼠中具有良好的耐受性、有利的理化性質(zhì)以及良好的口服生物利用度。在嚙齒動(dòng)物中, Nazartinib的分布量適中，清除率從低到中等（分別為大鼠和小鼠肝臟血流的30%和35%）。在狗中，Nazartinib表現(xiàn)出高清除率和高分布量。此外，Nazartinib在exon 20插入突變模型中顯示出抗腫瘤活性。在高于有效劑量的水平上，Nazartinib治療對(duì)WT EGFR的抑制作用最小且有良好的耐受性。在單劑量研究中，Nazartinib能持續(xù)抑制EGFR的磷酸化，這與其不可逆結(jié)合的能力相一致。Nazartinib在人體中的半衰期比在小鼠中更長(zhǎng)，目前正在進(jìn)行I/II期臨床試驗(yàn)，評(píng)估其在攜帶EGFR突變（包括T790M）的患者中的效果。 |
| 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 : 92 mg/mL (185.9 mM)
DMSO : 92 mg/mL (185.9 mM)
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | EGFR | NVS 816 | inhibit | HER1 | Epidermal growth factor receptor | NVS816 | EGF 816 | ErbB-1 | Inhibitor | Nazartinib | EGF-816 |
| 相關(guān)產(chǎn)品 | Lapatinib | Neratinib | Chalcone | Gefitinib | Erlotinib | Osimertinib | Erlotinib hydrochloride | Afatinib Dimaleate | Lidocaine Hydrochloride hydrate | Genistein | Khellin | Osimertinib mesylate |
| 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 | 膜蛋白靶向化合物庫 | 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 |