Pazopanib 是一种小分子抑制剂，可抑制多种具有潜在抗肿瘤活性的蛋白酪氨酸激酶。它抑制VEGFR1、VEGFR2、VEGFR3、PDGFRβ、c-Kit、FGFR1和c-Fms的IC50分别为10、30、47、84、74、140和146 nM。

Pazopanib, a small molecule inhibitor, inhibits multiple protein tyrosine kinases with potential antineoplastic activity. Pazopanib selectively inhibits VEGFR-1, -2 and -3, c-kit and PDGF-R, which may result in inhibition of angiogenesis in tumors in which these receptors are upregulated.

小鼠对Pazopanib治疗具有良好的耐受性,且每组小鼠的体重未出现差异.与低剂量组（载体或10 mg/kg Pazopanib）小鼠相比,高剂量组（30 mg/kg或100 mg/kg Pazopanib）小鼠肿瘤负荷明显降低.

在包括SYO-1和HS-SY-II细胞的所有滑膜肉瘤细胞系中,Pazopanib均呈现剂量生长依赖的抑制作用。Pazopanib浓度为1μg/ml时,SYO-1和HS-SY-II细胞的增殖被抑制,浓度为5μ时被完全抑制。在VEGF诱导的HUVEC 细胞中,Pazopanib能有效抑制VEGFR2磷酸化作用（IC50：8 nM）。Pazopanib由于引起G1期阻滞从而抑制滑膜肉瘤细胞生长。和载体处理的细胞不同,经Pazopanib处理的SYO-1细胞中的Akts,GSK-3β,JNKs,p70 S6 激酶,和mTOR磷酸化作用受到抑制。浓度在20 mg/ml和22.5 mg/ml之间时,Pazopanib浓度逐渐增加,RPE细胞活性降低。

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].

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].

444731-52-6

C21H23N7O2S

437.52

帕唑帕尼;GW786034

DMSO:81 mg/mL (185.1 mM)
H2O:<1 mgml
Ethanol:<1 mgml
Powder: -20°C for 3 years
In solvent: -80°C for 2 years