| 名稱(chēng) | 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. |
| 細(xì)胞實(shí)驗(yàn) | 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]. |
| 激酶實(shí)驗(yàn) | 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]. |
| 體外活性 | 方法：人非小細(xì)胞肺癌細(xì)胞 PC-9 (exon 19del)、H3255 (L858R)、PC-9ER (exon 19del+T790M) 和 H1975 (L858R+T790M) 用 Osimertinib (0.0001-10 μmol/L) 處理 72 h，使用 MTS 方法檢測(cè)細(xì)胞生長(zhǎng)抑制情況。
結(jié)果：Osimertinib 劑量依賴(lài)誘導(dǎo) PC-9、H3255、PC-9ER 和 H1975 細(xì)胞生長(zhǎng)，IC50 分別為 41、26、41 和 31?nM。[1]
方法：EGFR 突變的人非小細(xì)胞肺癌細(xì)胞 PC-9、H1975、H1650 和 H3255 用 Osimertinib (0.1-1000 nM) 處理 6 h，使用 Western Blot 方法檢測(cè)靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：Osimertinib 抑制 EGFR 突變腫瘤細(xì)胞的 pEGFR(Y1068)、pERK(P-p44/42)、pAKT(S473)。[2]
方法：人非小細(xì)胞肺癌細(xì)胞 NCI-H1975 用 Osimertinib (10-100 nM) 處理 24 h，然后用 2-20 Gy 的劑量照射，使用 Flow Cytometry 方法分析細(xì)胞周期情況。
結(jié)果：聯(lián)合治療組 G2/M 和 S 期細(xì)胞比例呈劑量依賴(lài)性降低。Osimertinib 減少照射后 G2/M 期細(xì)胞周期停滯。[3] |
| 體內(nèi)活性 | 方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Osimertinib (5-10 mg/kg) 口服給藥給攜帶人肺癌腫瘤 H1975、PC9 和 A431 的 SCID 小鼠，每天一次，持續(xù)七天。
結(jié)果：Osimertinib 治療顯著抑制 H1975、PC9 和 A431 腫瘤的生長(zhǎng)，表明在體內(nèi)具有抗腫瘤活性。[4]
方法：為檢測(cè)體內(nèi)抗腫瘤活性，將 Osimertinib (6 mg/kg) 口服給藥給用 PC-9/ffluc 細(xì)胞構(gòu)建軟腦膜癌病 (LMC) 模型的 SHO-SCID 小鼠，每天一次，持續(xù)五十天。
結(jié)果：Osimertinib 治療明顯延緩了 LMC 的發(fā)展。第三代 EGFR-TKI Osimertinib 可能是由 EGFR 突變型腫瘤引起的第一代或第二代 EGFR-TKI 耐藥性 LMC 的有效治療方法。[5] |
| 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 : 22 mg/mL (44 mM)
DMSO : 55 mg/mL (110.09 mM)
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 9.2 mg/mL (18.41 mM), Suspension. Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | AZD 9291 | Inhibitor | Ba/F3 cells | Osimertinib | H1975 | EGFR | PC-9 | tumor xenograft model | ErbB-1 | AZD9291 | HER1 | inhibit | cancer | Epidermal growth factor receptor |
| 相關(guān)產(chǎn)品 | Lapatinib | Neratinib | Chalcone | Gefitinib | Erlotinib | Erlotinib hydrochloride | Afatinib Dimaleate | Lidocaine Hydrochloride hydrate | BLU-945 | Genistein | Khellin | Osimertinib mesylate |
| 相關(guān)庫(kù) | 抗癌上市藥物庫(kù) | 抗癌活性化合物庫(kù) | EMA 上市藥物庫(kù) | 激酶抑制劑庫(kù) | 高選擇性抑制劑庫(kù) | FDA 上市藥物庫(kù) | 酪氨酸激酶分子庫(kù) | FDA 上市激酶抑制劑庫(kù) | 藥物功能重定位化合物庫(kù) | 抗癌臨床化合物庫(kù) |