甲苯磺酸索拉非尼 T0093

Product Introduction
Bioactivity


英文名: Sorafenib tosylate

描述: Sorafenib tosylate (Bay 43-9006) 是一种口服活性Raf 抑制剂，也是铁死亡激动剂。它是多激酶抑制剂，对VEGFR2，VEGFR3，PDGFRβ，FLT3和c-Kit 的IC50分别为 90 nM，15 nM，20 nM，57 nM 和 58 nM。它诱导细胞自噬和凋亡，有抗肿瘤活性。

细胞实验: Tumor cell lines were plated at 2 × 105 cells per well in 12-well tissue culture plates in DMEM growth media (10% heat-inactivated FCS) overnight. Cells were washed once with serum-free media and incubated in DMEM supplemented with 0.1% fatty acid-free BSA containing various concentrations of BAY 43-9006 in 0.1% DMSO for 120 minutes to measure changes in basal pMEK 1/2, pERK 1/2, or pPKB. Cells were washed with cold PBS (PBS containing 0.1 mmol/L vanadate) and lysed in a 1% (v/v) Triton X-100 solution containing protease inhibitors. Lysates were clarified by centrifugation, subjected to SDS-PAGE, transferred to nitrocellulose membranes, blocked in TBS-BSA, and probed with anti-pMEK 1/2 (Ser217/Ser221; 1:1000), anti-MEK 1/2, anti-pERK 1/2 (Thr202/Tyr204; 1:1000), anti-ERK 1/2, anti-pPKB (Ser473; 1:1000), or anti-PKB primary antibodies. Blots were developed with horseradish peroxidase (HRP)-conjugated secondary antibodies and developed with Amersham ECL reagent on Amersham Hyperfilm [1].

激酶实验: 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 the 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.

动物实验: Female NCr-nu/nu mice (Taconic Farms, Germantown, NY) were used for all studies. Three to five million cells were injected s.c. into the right flank of each mouse. DLD-1 tumors were established and maintained as a serial in vivo passage of s.c. fragments (3 × 3 mm) implanted in the flank using a 12-gauge trocar. A new generation of the passage was initiated every three weeks, and studies were conducted between generations 3 and 12 of this line. Treatment was initiated when tumors in all mice in each experiment ranged in size from 75 to 144 mg for antitumor efficacy studies and from 100 to 250 mg for studies of microvessel density and ERK phosphorylation. All treatment was administered orally once daily for the duration indicated in each experiment.

体外活性: 除 Raf-1 外，Sorafenib 还能在生化试验中抑制 VEGFR-3（IC50：20 nM）、BRAF wt（IC50：22 nM）、B-RAF V599E（IC50：38 nM）、VEGFR-2（IC50：90 nM）、PDGFR-β（IC50：57 nM）、c-KIT（IC50：68 nM）和 Flt3（IC50：58 nM）[1]。当 10-0505 细胞与抗人源抗 HGF 抗体共同处理时，Sorafenib 诱导的 c-Met、p70S6K 和 4EBP1 磷酸化显著降低，这表明Sorafenib 治疗会导致 HGF 分泌增加和c-Met 和 mTOR 靶点激活[2]。

体内活性: Sorafenib Tosylate（10、30、50及100 mg/kg，口服）依剂量依赖方式抑制06-0606及10-0505异种移植瘤生长（P<0.01）。Sorafenib同样显著降低了06-0606和10-0505异种移植瘤的生长速率。经Sorafenib（50/100 mg/kg）处理的小鼠中，06-0606瘤体重分别约为对照组的13%及5%。50 mg剂量的Sorafenib在带有5-1318、26-1004和10-0505细胞系的小鼠中显著抑制肿瘤生长（P<0.01）。在50 mg剂量下，06-0606、26-1004、5-1318和10-0505异种移植瘤中，Sorafenib和安慰剂处理瘤体末次治疗中位重量（mg）的T/C比分别是0.13、0.10、0.12和0.49 [2]。存活率在二乙基亚硝胺（DENA）组为73.3%，Sorafenib组为83.3%，相较之下正常对照组为100%。DENA组较正常对照组显著增加肝脏指数（增加1.51倍，p<0.05），而Sorafenib处理则与DENA组相比显著降低肝脏指数（p<0.05），Sorafenib组的肝脏指数显著降至低于正常对照组水平 [3]。

存储条件: Powder: -20°C for 3 years | In solvent: -80°C for 1 year

溶解度: DMSO : 50 mg/mL (78.49 mM)
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
H2O : < 1 mg/mL (insoluble or slightly soluble)


关键字: inhibit | CD135 | Sorafenib | FLT3 | Raf kinases | Sorafenib Tosylate | Cluster of differentiation antigen 135 | Sorafenib tosylate | VEGFR | Autophagy | Ferroptosis | Raf | Inhibitor | Vascular endothelial growth factor receptor | Apoptosis | Fms like tyrosine kinase 3

相关产品: Ara-SH | dl-Maackiain | Euphorbia Factor L2 | 2-Hydroxychalcone | AEE788 | 2-Methoxyestradiol | OsMundacetone | Ac-DEVD-CHO | Oleuropein | ASP3026

相关库: Drug Repurposing Compound Library | FDA-Approved Kinase Inhibitor Library | Anti-Cancer Drug Library | Membrane Protein-targeted Compound Library | Kinase Inhibitor Library | Inhibitor Library | Anti-Cancer Clinical Compound Library | Tyrosine Kinase Inhibitor Library | Anti-Cancer Active Compound Library | Anti-Cancer Approved Drug Library

甲苯磺酸索拉非尼 T0093信息由TargetMol中国为您提供，如您想了解更多关于甲苯磺酸索拉非尼 T0093报价、型号、参数等信息，欢迎来电或留言咨询。

注：该产品未在中华人民共和国食品药品监督管理部门申请医疗器械注册和备案，不可用于临床诊断或治疗等相关用途