印度多吉美

产品描述

Sorafenib is a potent multikinase inhibitor (IC50s: 6/20/22 nM for Raf-1/VEGFR-4/B-Raf).

靶点活性

B-Raf,22nM

B-Raf (V599E),38nM

Raf-1,6nM

VEGFR2/Flk1,15nM

VEGFR3,20nM

体外活性

Sorafenib tosylate明显抑制NIH 3T3细胞中VEGFR2磷酸化和HEK-293细胞中Flt-3磷酸化。Sorafenib tosylate也能有效抑制mVEGFR2 (Flk-1)，mVEGFR3，mPDGFRβ，Flt3和c-Kit，IC50分别为15 nM，20 nM，57 nM，58 nM和68 nM。Sorafenib tosylate抑制野生型和V599E突变型B-Raf活性，IC50分别为22 nM和38 nM。Sorafenib tosylate抑制HAoSMC和MDA-MB-231细胞的增殖，IC50分别为0.28 μM和2.6 μM。Sorafenib tosylate除了抑制RAF/MEK/ERK信号通路，明显抑制eIF4E的磷酸化作用，并以MEK/ERK依赖的方式降低肝癌的细胞中Mcl-1水平。Sorafenib tosylate抑制PLC/PRF/5和HepG2细胞的增殖，IC50分别为6.3 μM和4.5 μM，并诱导细胞凋亡。

体内活性

30-60 mg/kg Sorafenib与5 mg/kg TRAIL结合对TRAIL抗性HCT116 bax-/-和HT29肿瘤异种移植物有明显的功效。口服60 mg/kg Sorafenib tosylate ，对人肿瘤异种移植模MDA-MB-231，Colo-205，HT-29，DLD-1，NCI-H460和A549表现出广谱的、剂量依赖性抗肿瘤活性，并且没有毒性。Sorafenib通过下调NF-κB介导的Mcl-1 和 cIAP2的表达使bax-/-细胞对TRAIL剂量依赖性敏感。Sorafenib tosylate有效抑制HT-29 和 MDA-MB-231异种移植物中MEK 1/2磷酸化和pERK 1/2水平，并且明显抑制MDA MB-231，HT-29和Colo-205肿瘤异种移植物中肿瘤微血管面积和微血管密度。

激酶实验

Biochemical assays: Recombinant baculoviruses expressing Raf-1 (residues 305–648) and B-Raf (residues 409–765) are purified as fusion proteins. Full-length human MEK-1 is generated by PCR and purified as a fusion protein from Escherichia coli lysates. Sorafenib tosylate is added to a mixture of Raf-1 (80 ng), or B-Raf (80 ng) with MEK-1 (1 μg) in assay buffer [20 mM Tris (pH 8.2), 100 mM NaCl, 5 mM MgCl2, and 0.15% β-mercaptoethanol] at a final concentration of 1% DMSO. The Raf kinase assay (final volume of 50 μL) is initiated by adding 25 μL of 10 μM γ[33P]ATP (400 Ci/mol) and incubated at 32 °C for 25 minutes. Phosphorylated MEK-1 is harvested by filtration onto a phosphocellulose mat, and 1% phosphoric acid is used to wash away unbound radioactivity. After drying by microwave heating, a β-plate counter is used to quantify filter-bound radioactivity. Human VEGFR2 (KDR) kinase domain is expressed and purified from Sf9 lysates. Time-resolved fluorescence energy transfer assays for VEGFR2 are performed in 96-well opaque plates in the time-resolved fluorescence energy transfer format. Final reaction conditions are as follows: 1 to 10 μM ATP, 25 nM poly GT-biotin, 2 nM Europium-labeled phospho (p)-Tyr antibody (PY20), 10 nM APC, 1 to 7 nM cytoplasmic kinase domain in final concentrations of 1% DMSO, 50 mM HEPES (pH 7.5), 10 mM MgCl2, 0.1 mM EDTA, 0.015% Brij-35, 0.1 mg/mL BSA, and 0.1% β-mercaptoethanol. Reaction volumes are 100 μL and are initiated by addition of enzyme. Plates are read at both 615 and 665 nM on a Perkin-Elmer VictorV Multilabel counter at ~1.5 to 2.0 hours after reaction initiation. Signal is calculated as a ratio: (665 nm/615 nM) × 10,000 for each well. For IC50 generation, Sorafenib tosylate is added before the enzyme initiation. A 50-fold stock plate is made with Sorafenib tosylate serially diluted 1:3 in a 50% DMSO/50% distilled water solution. Final Sorafenib tosylate concentrations range from 10 μM to 4.56 nM in 1% DMSO.

细胞实验

Cells are exposed to increasing concentrations of Sorafenib tosylate for 72 hours. Cell number is quantitated using the Cell TiterGlo ATP Luminescent assay kit. This assay measures the number of viable cells per well by measurement of luminescent signal based on amount of cellular ATP. (Only for Reference)

细胞系： MDA-MB-231, and HAoSMC

动物实验

动物模型：FeNCr-nu/nu mice implanted s.c. with MDA-MB-231, Colo-205, HT-29, H460, or A549 cells

分子量

464.83

分子式

C21H16ClF3N4O3

CAS

284461-73-0

溶解度

DMSO: 59 mg/mL (126.9 mM)

Ethanol: <1 mg/mL

Water: <1 mg/mL

（ < 1 mg/ml refers to the product slightly soluble or insoluble )

储存条件

store at -80°C

备注

For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.