| 名稱 | AEE788 |
| 描述 | AEE788 (NVP-AEE 788) has been used in trials studying the treatment of Cancer, Glioblastoma Multiforme, and Brain and Central Nervous System Tumors. |
| 細胞實驗 | Methylene Blue Cell Proliferation Assay.Cells are seeded at 1.5 × 103 cells/well into 96-well microtiter plates and incubated overnight at 37 °C, 5% v/v CO2 and 80% relative humidity. AEE788 dilutions are added on day 1, with the highest concentration being 10 μM. After incubation of the cell plates for an additional 4 (T24) or 6 (BT-474, SK-BR-3, and NCI-H596) days, cells are fixed with 3.3% v/v glutaraldehyde, washed with water, and stained with 0.05% w/v methylene blue. After washing, the dye is eluted with 3% HCl and the absorbance measured at 665 nm with a SpectraMax 340 spectrophotometer. IC50 values are determined by mathematical curve-fitting and are defined as the drug concentration leading to 50% inhibition of net cell mass increase compared with untreated control cultures. (Only for Reference) |
| 激酶實驗 | Protein Kinase Assays: The in vitro kinase assays are performed in 96-well plates (30 μL) at ambient temperature for 15–45 min using the recombinant glutathione S-transferase-fused kinase domains (4-100 ng, depending on specific activity). [γ33P]ATP is used as phosphate donor and polyGluTyr-(4:1) peptide as acceptor. With the exception of protein kinase C-α, cyclin-dependent kinase 1/cycB and protein kinase A are protamine sulfate (200 μg/mL), histone H1 (100 μg/mL), and the heptapeptide Leu-Arg-Arg-Ala-Ser-Leu-Gly (known as Kemptide Bachem) respectively and are used as peptide substrates. Assays are optimized for each kinase using the following ATP concentrations: 1.0 μM (c-Kit, c-Met, c-Fms, c-Raf-1, and RET), 2.0 μM (EGFR, erbB2, ErbB3, and ErbB4), 5.0 μM (c-abl), 8.0 μM (Flt-1, Flt-3, Flt-4, Flk, KDR, FGFR-1, and Tek), 10.0 μM (PDGFR-β, protein kinase C-α, and cyclin-dependent kinase 1), and 20.0 μM (c-Src and protein kinase A). The reaction is terminated by the addition of 20 μL 125 mM EDTA. Thirty μL (c-abl, c-Src, insulin-like growth factor-1R, RET-Men2A, and RET-Men2B) or 40 μL (all other kinases) of the reaction mixture is transferred onto Immobilon-polyvinylidene difluoride membrane, presoaked with 0.5% H3PO4 and mounted on a vacuum manifold. Vacuum is then applied and each well rinsed with 200 μL 0.5% H3PO4. Membranes are removed and washed four times with 1.0% H3PO4 and once with ethanol. Dried membranes are counted after mounting in a Packard TopCount 96-well frame and with the addition of 10 μL/well of Microscint. IC50 values (±SE) are calculated by linear regression analysis of the percentage inhibition and are averages of at least three determinations. |
| 體外活性 | AEE788（50 mg/kg）抑制盲腸和腹膜腫瘤的生長（> 50％）,并將植入裸鼠盲腸的HT29細胞的淋巴結(jié)轉(zhuǎn)移發(fā)生率降低至70％,對體重無影響.50 mg/kg AEE788作用于NeuT/ErbB2 GeMag模型誘導腫瘤消退達57%.AEE788有效抑制A431腫瘤中的EGF誘導的EGFR磷酸化和GeMag腫瘤中的erbB2磷酸化.AEE788在NCI-H596或DU145異種移植模型中產(chǎn)生對腫瘤生長的劑量依賴性抑制,僅有輕微的體重變化.AEE788作用于K562腫瘤細胞,可以促進LBH589調(diào)節(jié)的活性氧簇的產(chǎn)生,可以增強凋亡. AEE788劑量依賴性地抑制由VEGF誘導的血管生成并且不抑制bFGF誘導的血管生成.AEE788作用于HT29盲腸癌,明顯降低pEGFR和pVEGFR 的表達水平,但是不改變EGF,VEGF,EGFR或VEGFR的表達水平.和CPT-11聯(lián)用,AEE788明顯抑制淋巴癌轉(zhuǎn)移 AEE788抑制Daoy,DaoyPt和DaoyHER2移植瘤的生長,抑制分別達51%,45%和72%. |
| 體內(nèi)活性 | 0.2-1.0 μM AEE788抑制HT29細胞中EGFR和Akt的磷酸化。AEE788抑制人皮膚SCC細胞系（Colo16,HaCaT,SRB1和SRB12細胞）中EGFR,VEGFR2,Akt和MAPK的磷酸化,導致生長抑制和誘導細胞凋亡。 AEE788有效抑制A431細胞中的EGFR磷酸化,IC5??0為11 nM。 AEE788還抑制CHO細胞中KDR的磷酸化和BT-474細胞中的erbB2的磷酸化,而對A31細胞中的PDGF-誘導的磷酸化沒有任何影響。AEE788作用于髓母細胞瘤細胞系,抑制細胞增殖,抑制EGF和神經(jīng)調(diào)節(jié)蛋白誘導的HER1,HER2和HER3激活。AEE788抑制NCI-H596,MK,BT-474和SK-BR-3細胞增殖,IC50分別為78,56,49和381 nM。AEE788也抑制EGF和VEGF促進的人臍靜脈內(nèi)皮細胞增殖,IC50分別為43和155 nM。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 82 mg/mL (186.1 mM)
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| 關(guān)鍵字 | Apoptosis | HER1 | Inhibitor | Epidermal growth factor receptor | AEE788 | NVP-AEE788 | ErbB-1 | inhibit | NVP-AEE-788 | AEE-788 | AEE 788 | EGFR |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Gefitinib | Metronidazole | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Myricetin | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 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 | 藥物功能重定位化合物庫 | 酪氨酸激酶分子庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |