| 名稱 | Pazopanib |
| 描述 | Pazopanib (GW786034) is an inhibitor of protein tyrosine kinases that inhibits VEGFR1, VEGFR2, VEGFR3, PDGFRβ, c-Kit, FGFR1, and c-Fms (IC50=10/30/47/84/74/140/146 nM). Pazopanib has antitumor activity. |
| 細(xì)胞實(shí)驗(yàn) | Pazopanib is prepared in DMSO and then diluted to final concentration in medium[1]. The effect of Pazopanib on cell proliferation is measured using 5-bromo-2-deoxyuridine (BrdU) incorporation method using commercially available kits. HUVEC is seeded in medium containing 5% fetal bovine serum (FBS) in type 1 collagen coated 96-well plates and incubated overnight at 37°C, 5% CO2. The medium is aspirated from the cells, and various concentrations of Pazopanib in serum-free medium are added to each well. After 30 min, either VEGF (10 ng/mL) or bFGF (0.3 ng/mL) is added to the wells. Cells are incubated for an additional 72 h and BrdU (10 μM) is added during the last 18 to 24 h of incubation. At the end of incubation, BrdU incorporation in cells is measured by ELISA. Data are fitted with a curve described by the equation, y=Vmax(1?(x/(K+x))), where K is equal to the IC50[1]. |
| 激酶實(shí)驗(yàn) | VEGFR enzyme assays for VEGGR1, VEGFR2, and VEGFR3 are run in homogeneous time-resolved fluorescence (HTRF) format in 384-well microtiter plates using a purified, baculovirus-expressed glutathione-S-transferase (GST) fusion protein encoding the catalytic c-terminus of human VEGFR receptor kinases 1, 2, or 3. Reactions are initiated by the addition of 10 μL of activated VEGFR2 kinase solution [final concentration, 1 nM enzyme in 0.1 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 7.5, containing 0.1 mg/mL bovine serum albumin (BSA), 300 μM dithiothreitol (DTT)] to 10 μL substrate solution [final concentration, 360 nM peptide, (biotin-aminohexyl-EEEEYFELVAKKKK-NH2), 75 μM ATP, 10 μM MgCl2], and 1 μL of titrated compound in DMSO. Plates are incubated at room temperature for 60 min, and then the reaction is quenched by the addition of 20 μL of 100 mM ethylene diamine tetraacetic acid (EDTA). After quenching, 20 μL HTRF reagents (final concentration, 15 nM Streptavidin-linked allophycocyanin, 1 nM Europium-labeled antiphosphotyrosine antibody diluted in 0.1 mg/mL BSA, 0.1 M HEPES, pH 7.5) is added and the plates incubated for a minimum of 10 min. The fluorescence at 665 nM is measured with a Wallac Victor plate reader using a time delay of 50 μs[1]. |
| 體外活性 | 方法: SCLC 細(xì)胞系 NCI-H446 和 NCI-H82 用 Pazopanib (0.01-30 μM) 處理 24-72 h，通過 CCK-8 assay 檢測(cè)細(xì)胞活力。
結(jié)果: Pazopanib 以劑量和時(shí)間依賴的方式顯著降低了 NCI-H446 細(xì)胞的增殖，24 h 時(shí) IC50 值為 1.05 μM。Pazopanib 還可以以劑量和時(shí)間依賴的方式誘導(dǎo) NCI-H82 細(xì)胞中的強(qiáng)效細(xì)胞死亡，24 h 的 IC50 為 1.298 μM。Pazopanib 顯著拮抗小細(xì)胞肺癌細(xì)胞增殖。[1]
方法: 人結(jié)直腸癌細(xì)胞 HCT-116 用 Pazopanib (1-20 μM) 處理 3-24 h，通過 Western Blot 檢測(cè)靶點(diǎn)蛋白表達(dá)水平。
結(jié)果: Pazopanib 顯著誘導(dǎo)了 PUMA 的表達(dá)，這是時(shí)間和劑量依賴性的。[2] |
| 體內(nèi)活性 | 方法: 為檢測(cè)體內(nèi)抗腫瘤活性，將 Pazopanib (30 mg/kg，suspended in 0.5% hydroxypropylmethyl cellulose and 0.1% Tween-80 in water) 灌胃給藥給攜帶 NCI-H446 異種移植物的 NOD-SCID 小鼠，每天一次，持續(xù)兩周。
結(jié)果: Pazopanib 的給藥顯著抑制了 NCI-H446 異種移植物的生長(zhǎng)。[1] |
| 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 : < 1 mg/mL (insoluble or slightly soluble)
H2O : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 20 mg/mL (45.71 mM)
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| 關(guān)鍵字 | FGFR | Autophagy | Platelet-derived growth factor receptor | Pazopanib | GW 786034 | c-Kit | PDGFR | Vascular endothelial growth factor receptor | CD117 | inhibit | VEGFR | Fibroblast growth factor receptor | GW-786034 | Inhibitor | SCFR |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | Taurine | Gefitinib | Aceglutamide | Hydroxychloroquine | Ferulic Acid | Curcumin | Stavudine | Paeonol | Sodium 4-phenylbutyrate |
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