| 名稱 | Erlotinib hydrochloride |
| 描述 | Erlotinib hydrochloride (NSC 718781) is an EGFR inhibitor (IC50: 2 nM). It is used for the treatment of non-small cell lung cancer. |
| 細(xì)胞實(shí)驗(yàn) | Exponentially growing cells are seeded in 96-well plastic plates and exposed to serial dilutions of erlotinib (30 nM-20 μM), pemetrexed, or the combination at a constant concentration ratio of 4:1 in triplicates for 72 h. Cell viability is assayed by cell count and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Growth inhibition is expressed as the percentage of surviving cells in drug-treated versus PBS-treated control cells (which is considered as 100% viability). The IC50 value is the concentration resulting in 50% cell growth inhibition by a 72-h exposure to the drug(s) compared with untreated control cells and is calculated by the CalcuSyn software [4]. |
| 激酶實(shí)驗(yàn) | 96-well plates are coated by incubation overnight at 37 °C with 100 μL per well of 0.25 mg/mL PGT in PBS. Excess PGT is removed by aspiration, and the plate is washed 3 times with washing buffer (0.1% Tween 20 in PBS). The kinase reaction is performed in 50 μL of 50 mM HEPES (pH 7.3), containing 125 mM sodium chloride, 24 mM magnesium chloride, 0.1 mM sodium orthovanadate, 20 μM ATP, 1.6 μg/mL EGF, and 15 ng of EGFR, affinity purified from A431 cell membranes. Erlotinib HCl in DMSO is added to give a final DMSO concentration of 2.5%. Phosphorylation is initiated by addition of ATP and proceeded for 8 minutes at room temperature, with constant shaking. The kinase reaction is terminated by aspiration of the reaction mixture and is washed 4 times with washing buffer. Phosphorylated PGT is measured by 25 minutes of incubation with 50 μL per well HRP-conjugated PY54 anti-phosphotyrosine antibody, diluted to 0.2 μg/mL in blocking buffer (3% BSA and 0.05% Tween 20 in PBS). The antibody is removed by aspiration, and the plate is washed 4 times with washing buffer. The colorimetric signal is developed by addition of TMB Microwell Peroxidase Substrate, 50μL per well, and stopped by the addition of 0.09 M sulfuric acid, 50 μL per well. Phosphotyrosine is estimated by measurement of absorbance at 450 nm. The signal for controls is typically 0.6-1.2 absorbance units, with essentially no background in wells without AlP, EGFR, or PGT and is proportional to the time of incubation for 10 minutes [1]. |
| 動(dòng)物實(shí)驗(yàn) | Six-week-old male SD rats weighing 180 to 210 g are used. Cisplatin (CP) is freshly prepared in saline at a concentration of 1 mg/mL and then injected intraperitoneally in SD rats (n=28) at a dose of 7 mg/kg on day 0. To investigate the effect of Erlotinib, 28 CP-N rats are divided into two groups. Separate groups (n=14) each of animals are administered with either Erlotinib (20 mg/kg) (CP+E, n=14) or vehicle (CP+V, n=14) daily by oral gavage from the day -1 (24 hours prior to the CP injection) to day 3. Vehicle-treated groups receive an equivalent volume of saline. Five male SD rats at the age of 6 weeks are used as a normal control group (NC, n=5). The NC rats are given an equivalent volume of saline daily by oral gavage from the day -1 to day 3. At day 4 (96 hours after CP injection), each rat is anesthetized and sacrificed by exsanguination after the cardiac puncture; blood is collected by cardiac puncture and kidneys are collected. Renal tissue is divided; separate portions are snap-frozen in liquid nitrogen or fixed in 2% paraformaldehyde/phosphate-buffered saline (PBS) for later use. All surgery is performed under diethyl ether gas anesthesia, and all efforts are made to minimize suffering [2]. |
| 體外活性 | Erlotinib 是一種直接作用于人類EGFR酪氨酸激酶的抑制劑（IC50為2nM），能夠顯著降低完整腫瘤細(xì)胞中EGFR的自磷酸化作用（IC50為20nM）。同時(shí)，Erlotinib 對(duì)重組的EGFR細(xì)胞內(nèi)（激酶）域也是一種有效抑制劑（IC50為1nM）。DiFi細(xì)胞的增殖受到Erlotinib 的強(qiáng)力抑制，在為期8天的增殖試驗(yàn)中，IC50達(dá)到100nM[1]。 |
| 體內(nèi)活性 | Erlotinib（20 mg/kg，口服）顯著減輕順鉑(CP)所引起的大鼠體重(BW)損失，與CP+載體(V)組相比(P<0.05)。Erlotinib 治療明顯改善了CP-N（正常對(duì)照組，NC）大鼠的腎功能。與CP+V大鼠相比，CP+E（Erlotinib ）組大鼠的血清肌酐(s-Cr)水平、血尿素氮(BUN)、尿N-乙酰-β-D-葡萄糖胺酶(NAG)指數(shù)顯著降低（P<0.05），且尿量(UV)及肌酐清除率(Ccr)顯著提高（P<0.05）[2]。Erlotinib 在小鼠體內(nèi)顯著抑制人頭頸癌HN5腫瘤異種移植物的生長(zhǎng)，ED50值為9 mg/kg [3]。 |
| 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 : 5.5 mg/mL (12.79 mM)
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| 關(guān)鍵字 | OSI774 | Erlotinib hydrochloride | Inhibitor | EGFR | HER1 | inhibit | Epidermal growth factor receptor | Erlotinib | NSC718781 | OSI 744 | OSI-774 | OSI744 | OSI 774 | CP358774 | CP 358774 | Autophagy | Erlotinib Hydrochloride | ErbB-1 | NSC-718781 |
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