| 名稱 | Vandetanib |
| 描述 | Vandetanib (ZD6474) is a potent inhibitor of VEGFR2 (IC50: 40 nM). It also inhibits VEGFR3 and EGFR. |
| 細胞實驗 | HUVEC proliferation in the presence and absence of growth factors was evaluated using [3H]thymidine incorporation. Briefly, HUVECs isolated from umbilical cords were plated (at passage 2–8) in 96-well plates (1000 cells/well) and dosed with ZD6474 ± VEGF or EGF (3 ng/ml) or bFGF (0.3 ng/ml). The cultures were incubated for 4 days (37°C; 7.5% CO2) and then pulsed with 1 μCi/well [3H]thymidine and reincubated for 4 h. Cells were harvested and assayed for the incorporation of tritium using a beta counter. IC50 data were interpolated as described above [1]. |
| 激酶實驗 | The ability of ZD6474 to inhibit the kinase activity associated with the VEGFRs KDR, Flt-1, and Flt-4 was determined using a previously described ELISA. Briefly, ZD6474 was incubated with enzyme, 10 mm MnCl2, and 2 μm ATP in 96-well plates coated with a poly(Glu, Ala, Tyr) 6:3:1 random copolymer substrate. Phosphorylated tyrosine was then detected by sequential incubation with a mouse IgG anti-phosphotyrosine 4G10 antibody, a horseradish peroxidase-linked sheep anti-mouse immunoglobulin antibody, and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). Microcal Origin software was used to interpolate IC50 values by nonlinear regression. This methodology was adapted to examine selectivity versus tyrosine kinases associated with EGFR, PDGFRβ, Tie-2, FGFR1, c-kit, erbB2, IGF-IR, and FAK. All enzyme assays (tyrosine or serine-threonine) used appropriate ATP concentrations at or just below the respective Km (0.2–14 μm). Selectivity versus serine-threonine kinases (CDK2, AKT, and PDK1) was examined using a relevant scintillation proximity assay (SPA) in 96-well plates. CDK2 assays contained 10 mm MnCl2, 4.5 μm ATP, 0.15 μCi of [γ-33P]ATP/reaction, 50 mm HEPES (pH 7.5), 1 mm DTT, 0.1 mm sodium orthovanadate, 0.1 mm sodium fluoride, 10 mm sodium glycerophosphate, 1 mg/ml BSA fraction V, and a retinoblastoma substrate (part of the retinoblastoma gene, 792–928, expressed in a glutathione S-transferase expression system; 0.22 μm final concentration). Reactions were allowed to proceed at room temperature for 60 min before quenching for 2 h with 150 μl of a solution containing EDTA (62 mm final concentration), 3 μg of a rabbit immunoglobulin anti-glutathione S-transferase antibody and protein A SPA-polyvinyltoluene beads (0.8 mg/reaction). Plates were then sealed, centrifuged (1200 × g for 5 min), and counted on a Topcount NXT Microplate scintillation counter for 30 s [1]. |
| 動物實驗 | Methodology to enable blood pressure measurement in anesthetized rats was as described previously. Briefly, anesthesia was induced in male Alderley Park rats using α-chloralose by the i.v. route and then maintained with thiopentone via the i.p. route. Once surgical anesthesia was established, the carotid artery was cannulated to enable blood pressure recording using a pressure transducer and a Lectromed MT8P amplifier. The jugular vein was cannulated to allow growth factor administration. Body temperature was maintained with a thermostatically controlled heated blanket coupled to a rectal thermometer. Human VEGF165 (32 μg/kg) or bFGF (40 μg/kg) was administered as a bolus injection [0.1 ml/250 g body weight in 0.85% (w/v) sodium chloride], and a maximal blood pressure drop was recorded within 2 min (typically 26–30 mm Hg). These changes were sustainable for more than 20 min in control experiments. ZD6474 (2.5 mg/kg) or vehicle alone [25% (w/v) hydroxypropyl-β-cyclodextrin in Sorensons phosphate buffer (pH 5.5)] was administered i.v., and blood pressure was recorded 5 min later to determine the effect on growth factor-induced hypotension [1]. |
| 體外活性 | Vandetanib (ZD6474) 是一種強效的KDR/VEGFR2酪氨酸激酶活性抑制劑（IC50：40 nM），對VEGFR3（IC50：110 nM）和EGFR/HER1酪氨酸激酶活性（IC50：500 nM）也有一定抑制作用。ZD6474對KDR酪氨酸激酶的抑制作用能強效抑制VEGF激活的內(nèi)皮細胞（人臍靜脈內(nèi)皮細胞）增殖（IC50：60 nM）[1]。ZD6474能劑量依賴性抑制小鼠NIH-EGFR成纖維細胞和人MCF-10A ras乳腺癌細胞中EGFR的磷酸化，并在具有功能性EGFR但缺乏VEGFR-2的七種人類細胞系中，劑量依賴性抑制軟瓊脂生長。ZD6474與紫杉醇或多西他賽聯(lián)合治療在體外觀察到增長抑制和凋亡的劑量依賴性超加性效應[2]。Vandetanib和neratinib對基礎ABCG2-ATP酶活性表現(xiàn)出抑制效果，在相對高濃度（10-20 mM）時，vandetanib能抑制激活的ABCG2-ATP酶活性[3]。 |
| 體內(nèi)活性 | 給予ZD6474 (2.5 mg/kg, 靜脈注射) 能夠逆轉(zhuǎn)由VEGF引起的低血壓變化(63%)，但對由基礎成纖維細胞生長因子引起的變化無顯著影響。每日一次口服50 mg/kg ZD6474給缺乏胸腺的小鼠（這些小鼠體內(nèi)已經(jīng)皮下植入了A549腫瘤細胞）同樣顯著抑制了由腫瘤引起的新血管生成（5天后抑制率達63%）。對于用50 mg/kg/day ZD6474處理24天的Calu-6腫瘤進行組織學分析顯示，非壞死區(qū)域內(nèi)CD31（內(nèi)皮細胞）染色顯著減少（>70%）[1]。ZD6474對攜帶可觸知GEO結(jié)腸癌異種移植瘤的裸鼠治療顯示出劑量依賴性的腫瘤生長抑制作用。ZD6474在GEO腫瘤異種移植瘤模型中的抗腫瘤活性，在與紫杉醇聯(lián)合應用時得到增強。所有使用ZD6474加紫杉醇治療的小鼠觀察到腫瘤退縮，并且伴隨著顯著增強的抗血管生成抑制作用[2]。Vandetanib (15 mg/kg) 對H1650/PTEN和H1650親本異種移植瘤的生長具有相似的效果[4]。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 27.5 mg/mL (57.85 mM), Sonication is recommended.
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| 關鍵字 | VEGFR | inhibit | Vascular endothelial growth factor receptor | Inhibitor | Apoptosis | ZD-6474 | Autophagy | ZD 6474 | Vandetanib |
| 相關產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | L-Glutamic acid | Gefitinib | Hydroxychloroquine | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | L-Ascorbic acid | Paeonol | Sodium 4-phenylbutyrate |
| 相關庫 | 抗癌上市藥物庫 | 抗癌活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 藥物功能重定位化合物庫 | FDA 上市激酶抑制劑庫 |