| 名稱 | BMS-599626 |
| 描述 | BMS-599626 (AC480) has been used in trials studying the treatment of Cancer, Metastases, and HER2 or EGFR Expressing Advanced Solid Malignancies. |
| 細(xì)胞實(shí)驗(yàn) | All cell lines are maintained in RPMI 1640 supplemented with 10% fetal bovine serum, 100 units/mL penicillin, and 100 μg/mL streptomycin. Cells are plated at 1,000 per well in 96-well plates and are cultured for 24 hours before BMS-599626 is added. BMS-599626 is diluted in culture medium such that the final concentrations of DMSO are ≤ 1%. Following the addition of BMS-599626, the cells are cultured for an additional 72 hours before cell viability is determined by measuring the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye with the CellTiter96 kit. For some cell lines, there is a lack of a correlation between 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye metabolism and cell number, and a thymidine uptake assay is used to measure proliferation of these cell lines. Cells are plated in 96-well plates and treated with compounds as above. At the end of the 72-hour incubation, cells are pulsed with [3H]thymidine (0.4 μCi/well) for 3 hours before they are harvested. Cells are digested with 2.5% trypsin for 10 minutes at 37 °C and are harvested by filtration using a Packard Filtermate Harvester and GF/C Unifilter plates. Incorporation of radioactive thymidine into nucleic acids is determined by liquid scintillation counting.(Only for Reference) |
| 激酶實(shí)驗(yàn) | Protein kinase assays: The entire cytoplasmic sequences of HER1, HER2, and HER4 are expressed as recombinant proteins in Sf9 insect cells. HER1 and HER4 are expressed as fusion proteins with glutathione-S-transferase and are purified by affinity chromatography on glutathione-S-Sepharose. HER2 is subcloned into the pBlueBac4 vector and expressed as an untagged protein using an internal methionine codon (M687) for translation initiation. The truncated HER2 protein is isolated by chromatography on a column of DEAE-Sepharose equilibrated in a buffer that contains 0.1 M NaCl, and the recombinant protein is eluted with a buffer containing 0.3 M NaCl. For the HER kinase assays, reaction volumes are 50 μL and contains 10 ng of glutathione-S-transferase fusion protein or 150 ng of partially purified HER2. The mixtures also contains 1.5 μM poly(Glu/Tyr) (4:1), 1 μM ATP, 0.15 μCi [γ-33P]ATP, 50 mM Tris-HCl (pH 7.7), 2 mM DTT, 0.1 mg/mL bovine serum albumin, and 10 mM MnCl2. Reactions are allowed to proceed at 27°C for 1 hour and are terminated by the addition of 10 μL of a stop buffer (2.5 mg/mL bovine serum albumin and 0.3 M EDTA), followed by a 108-μL mixture of 3.5 mM ATP and 5% trichloroacetic acid. Acid-insoluble proteins are recovered on GF/C Unifilter plates with a Filtermate harvester. Incorporation of radioactive phosphate into the poly(Glu/Tyr) substrate is determined by liquid scintillation counting. Percent inhibition of kinase activity is determined by nonlinear regression analyses and data are reported as the inhibitory concentration required to achieve 50% inhibition relative to control reactions (IC50). Data are the averages of triplicate determinations. All other tyrosine kinases are also assayed using poly(Glu/Tyr) as a substrate. Kinetics of HER1 and HER2 inhibition are determined in reaction mixtures that contains varying concentrations of ATP and BMS-599626. |
| 體外活性 | 在體內(nèi),口服60 mg/kg到240 mg/kg范圍的BMS-599626,導(dǎo)致劑量依賴性抑制 Sal2腫瘤生長(zhǎng),在其人乳腺腫瘤KPL-4異種移植物中產(chǎn)生有效的抗腫瘤活性最大耐受劑量為180 mg/kg,并且在其他HER2擴(kuò)增異種移植模型以及其他HER1過(guò)表達(dá)異種移植模型中也具有類似的抗腫瘤活性. |
| 體內(nèi)活性 | MS-599626抑制表達(dá)高水平HER1和/或HER2的腫瘤細(xì)胞增殖,包括Sal2,BT474,N87,KPL-4,HCC202,HCC1954,HCC1419,AU565,ZR-75-30,MDA-MB-175,GEO和PC9細(xì)胞,IC50分別為0.24 μM,0.31 μM,0.45 μM,0.38 μM,0.94 μM,0.34 μM,0.75 μM,0.63 μM,0.51 μM,0.84 μM,0.90 μM和0.34 μM。BMS-599626選擇性抑制重組HER1和HER2激酶的酶活性,IC50分別為20 nM和30 nM。BMS-599626通過(guò)促進(jìn)周期重新分布和抑制DNA修復(fù),顯著增強(qiáng)了表達(dá)EGFR和Her2細(xì)胞的HN-5細(xì)胞的放射敏感性。此外,BMS-599626也抑制相關(guān)受體HER4,但是效力較低,IC50為190 nM。BBMS-599626不會(huì)明顯抑制不表達(dá)HER1或HER2的卵巢腫瘤細(xì)胞系A(chǔ)2780和MRC5成纖維細(xì)胞的增殖。 |
| 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 | H2O : <1 mg/mL
DMSO : 104 mg/mL (196 mM)
Ethanol : 16 mg/mL (30.2 mM)
|
| 關(guān)鍵字 | antitumor | Epidermal growth factor receptor | Sal2 | inhibit | cells | ErbB-1 | AC-480 | proliferation | EGFR | KPL4 | GEO | AC 480 | Inhibitor | HER1 | cancer | autophosphorylation | BMS 599626 | BMS-599626 | selective | BMS599626 |
| 相關(guān)產(chǎn)品 | Lidocaine hydrochloride | Lapatinib | Neratinib | Chalcone | Lidocaine | Gefitinib | Erlotinib | Osimertinib | Erlotinib hydrochloride | Lidocaine Hydrochloride hydrate | Genistein | Nintedanib |
| 相關(guān)庫(kù) | 抗卵巢癌化合物庫(kù) |