| 名称 | Osimertinib |
| 描述 | Osimertinib (AZD-9291) is an EGFR third-generation inhibitor that inhibits the T790M resistance mutation produced by second-generation EGFR inhibitors with irreversible and oral activity. Osimertinib has antitumor activity for the treatment of EGFR-mutated non-small-cell lung cancer. |
| 细胞实验 | AZD-9291 is dissolved in DMSO and stored, and then diluted with appropriate medium before use[1]. PC-9 cells are seeded into T75 flasks (5×105 cells/flask) in RPMI growth media and incubated at 37°C, 5% CO2. The following day the media is replaced with media supplemented with a concentration of EGFR inhibitor equal to the EC50 concentration predetermined in PC-9 cells. Media changes are carried out every 2-3 days and resistant clones allowed to grow to 80% confluency prior to the cells being trypsinised and reseeded at the original seeding density in media containing twice the concentration of EGFR inhibitor. Dose escalations are continued until a final concentration of 1.5 μM Gefitinib, 1.5 μM Afatinib, 1.5 μM WZ4002 or 160 nM AZD-9291 are achieved[1]. |
| 激酶实验 | ACY-1215 is dissolved and subsequently diluted in assay buffer [50 mM HEPES, pH 7.4, 100 mM KCl, 0.001% Tween-20, 0.05% BSA, and 20 μM tris(2-carboxyethyl)phosphine] to 6-fold the final concentration. HDAC enzymes are diluted to 1.5-fold of the final concentration in assay buffer and pre-incubated with ACY-1215 for 10 minutes before the addition of the substrate. The amount of FTS (HDAC1, HDAC2, HDAC3, and HDAC6) or MAZ-1675 (HDAC4, HDAC5, HDAC7, HDAC8, and HDAC9) used for each enzyme is equal to the Michaelis constant (Km), as determined by a titration curve. FTS or MAZ-1675 is diluted in assay buffer to 6-fold the final concentration with 0.3 μM sequencing grade trypsin. The substrate/trypsin mix is added to the enzyme/compound mix and the plate is shaken for 60 seconds and then placed into a SpectraMax M5 microtiter plate reader. The enzymatic reaction is monitored for release of 7-amino-4-methoxy-coumarin over 30 minutes, after deacetylation of the lysine side chain in the peptide substrate, and the linear rate of the reaction is calculated[1]. |
| 体外活性 | 方法：人非小细胞肺癌细胞 PC-9 (exon 19del)、H3255 (L858R)、PC-9ER (exon 19del+T790M) 和 H1975 (L858R+T790M) 用 Osimertinib (0.0001-10 µmol/L) 处理 72 h，使用 MTS 方法检测细胞生长抑制情况。
结果：Osimertinib 剂量依赖诱导 PC-9、H3255、PC-9ER 和 H1975 细胞生长，IC50 分别为 41、26、41 和 31 nM。[1]
方法：EGFR 突变的人非小细胞肺癌细胞 PC-9、H1975、H1650 和 H3255 用 Osimertinib (0.1-1000 nM) 处理 6 h，使用 Western Blot 方法检测靶点蛋白表达水平。
结果：Osimertinib 抑制 EGFR 突变肿瘤细胞的 pEGFR(Y1068)、pERK(P-p44/42)、pAKT(S473)。[2]
方法：人非小细胞肺癌细胞 NCI-H1975 用 Osimertinib (10-100 nM) 处理 24 h，然后用 2-20 Gy 的剂量照射，使用 Flow Cytometry 方法分析细胞周期情况。
结果：联合治疗组 G2/M 和 S 期细胞比例呈剂量依赖性降低。Osimertinib 减少照射后 G2/M 期细胞周期停滞。[3] |
| 体内活性 | 方法：为检测体内抗肿瘤活性，将 Osimertinib (5-10 mg/kg) 口服给药给携带人肺癌肿瘤 H1975、PC9 和 A431 的 SCID 小鼠，每天一次，持续七天。
结果：Osimertinib 治疗显著抑制 H1975、PC9 和 A431 肿瘤的生长，表明在体内具有抗肿瘤活性。[4]
方法：为检测体内抗肿瘤活性，将 Osimertinib (6 mg/kg) 口服给药给用 PC-9/ffluc 细胞构建软脑膜癌病 (LMC) 模型的 SHO-SCID 小鼠，每天一次，持续五十天。
结果：Osimertinib 治疗明显延缓了 LMC 的发展。第三代 EGFR-TKI Osimertinib 可能是由 EGFR 突变型肿瘤引起的第一代或第二代 EGFR-TKI 耐药性 LMC 的有效治疗方法。[5] |
| 存储条件 | In solvent: -80°C for 1 year | Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | Ethanol : 22 mg/mL (44.03 mM), Sonication is recommended.
DMSO : 120 mg/mL (240.19 mM), Sonication is recommended.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 9.2 mg/mL (18.41 mM), Suspension.
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 关键字 | tumor xenograft model | PC-9 | Osimertinib | Inhibitor | inhibit | HER1 | H1975 | ErbB-1 | Epidermal growth factor receptor | EGFR (L858R/T790M) | EGFR (Exon 19 deletion) | EGFR | cancer | Ba/F3 cells | AZD9291 | AZD 9291 |
| 相关产品 | Neratinib | Lapatinib | Chalcone | Gefitinib | Erlotinib | Erlotinib hydrochloride | Afatinib | Afatinib Dimaleate | Lidocaine Hydrochloride hydrate | Genistein | Khellin | (S)-Afatinib |
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