索拉非尼作用機制 - Medchemexpress - MCE中國

1、Product Data SheetSorafenibCat. No.: HY-10201CAS No.: 284461-73-0分式: CHClFNO分量: 464.83作靶點: Raf; VEGFR; FLT3; Autophagy; Apoptosis; Ferroptosis作通路: MAPK/ERK Pathway; Protein Tyrosine Kinase/RTK; Autophagy; Apoptosis儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 DMSO :

2、 45 mg/mL (96.81 mM)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制備儲備液1 mM 2.1513 mL 10.7566 mL 21.5132 mL5 mM 0.4303 mL 2.1513 mL 4.3026 mL10 mM 0.2151 mL 1.0757 mL 2.1513 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 mo

3、nth。-80C 儲存時，請在 6 個內(nèi)使，-20C 儲存時，請在 1 個內(nèi)使。體內(nèi)實驗請根據(jù)您的實驗動物和給藥式選擇適當?shù)娜芙獍?。以下溶解案都請先按?In Vitro 式配制澄清的儲備液，再依次添加助溶劑：為保證實驗結(jié)果的可靠性，澄 的儲備液可以根據(jù)儲存條件，適當保存；體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的式助溶1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.38

4、mM); Clear solution此案可獲得 2.5 mg/mL (5.38 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 400 L PEG300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (5.38 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.5

5、mg/mL (5.38 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.38 mM); Clear solution此案可獲得 2.5 mg/mL (5.38 mM，飽和度未知) 的澄 溶液，此案不適于實驗周 期在半個以上的實驗。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 油中，混合均勻

6、。BIOLOGICAL ACTIVITY物活性 Sorafenib (Bay 43-9006)種有效的服活性 Raf 抑制劑，對 Raf-1 和 B-Raf 的 IC50 分別為 6 nM 和 20 nM。Sorafenib 種多激酶抑制劑，對 VEGFR2，VEGFR3，PDGFR，F(xiàn)LT3 和 c-Kit 的 IC50 分別為 90 nM，15 nM，20nM，57 nM 和 58 nM。Sorafenib 誘導細胞噬 (autophagy) 和凋亡 (apoptosis)，并具有抗腫瘤活性。Sorafenib 也是種 ferroptosis 激動劑。IC & Target VEGFR3

7、Braf Raf-1 VEGFR220 nM (IC50) 22 nM (IC50) 6 nM (IC50) 90 nM (IC50)PDGFR BrafV599E c-Kit Flt357 nM (IC50) 38 nM (IC50) 68 nM (IC50) 58 nM (IC50)體外研究 Sorafenib (BAY 43-9006) also inhibits BRAFwt (IC50=22 nM), BRAFV599E (IC50=38 nM), VEGFR-2 (IC50=90 nM), VEGFR-3 (IC50=20 nM), PDGFR- (IC50=57 nM), c-K

8、IT (IC50=68 nM), and Flt3 (IC50=58 nM) in biochemical assays. In MDA-MB-231 breast cancer cells, Sorafenib completely blocks activation of the MAPK pathway. Cells are preincubated withSorafenib (0.01 to 3 M), and dose-dependent inhibition of basal MEK 1/2 and ERK 1/2 phosphorylation (IC50, 40 and100

9、 nM, respectively)1.體內(nèi)研究 Sorafenib demonstrates broad oral antitumor efficacy in panel of human tumor xenograft models. Sorafenib is givenorally at 7.5 to 60 mg/kg. There is no lethality and no increase in weight loss in any treated group relative to the corresponding control group. Daily oral admin

10、istration of Sorafenib (30 to 60 mg/kg) produces complete tumor stasis during treatment in five of the six models1. The survival rate is 73.3 % in Diethyl nitrosamine (DENA) group and 83.3% in Sorafenib group compared to 100 % in the normal control group. DENA group shows a significant increase inli

11、ver index (1.51-fold increase, p0.05) compared to normal control group, while treatment with Sorafenib showssignificant decrease (p0.05) in liver index when compared to DENA group. The liver index in Sorafenib groupsignificantly decreases to lower than its value in the normal control2.PROTOCOLKinase

12、 Assay 1 To test compound inhibition against various RAF kinase isoforms, Sorafenib is added to a mixture of Raf-1 (80 ng), wtBRAF, or V599E BRAF (80 ng) with MEK-1 (1 g) in assay buffer 20 mM Tris (pH 8.2), 100 mM NaCl, 5 mM MgCl2, and0.15% -mercaptoethanol at a final concentration of 1% DMSO. The

13、RAF kinase assay (final volume of 50 L) isinitiated by adding 25 L of 10 M -33PATP (400 Ci/mol) and incubated at 32C for 25 minutes. PhosphorylatedMEK-1 is harvested by filtration onto a phosphocellulose mat, and 1% phosphoric acid is used to wash awayunbound radioactivity. After drying by microwave

14、 heating, a -plate counter is used to quantify filter-boundradioactivity1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedChemECell Assay 1 The MDA-MB-231 human mammary adenocarcinoma cell lines are plated at 2105 cells per well in 12

15、-well tissueculture plates in DMEM growth media (10% heat-inactivated FCS) overnight. Cells are washed once with serum-freemedia and incubated in DMEM supplemented with 0.1% fatty acid-free BSA containing various concentrations of BAY43-9006 (0.01, 0.03 , 0.1, 0.3, 1, 3 M) in 0.1% DMSO for 120 minut

16、es to measure changes in basal pMEK 1/2, pERK1/2, or pPKB. Cells are washed with cold PBS (PBS containing 0.1 mM vanadate) and lysed in a 1% (v/v) Triton X-100solution containing protease inhibitors. Lysates are clarified by centrifugation, subjected to SDS, transferred tonitrocellulose membranes, b

17、locked 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. Blotsare developed with horseradish peroxidase (HRP)-conjugated secondary antibodies and developed w

18、ith AmershamECL reagent on Amersham Hyperfilm1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice1Administration 12 Female NCr-nu/nu mice are used. Mice bearing 75 to 150 mg tumors are treated orally with Sorafenib (7.5 to 60mg/kg), administere

19、d daily for 9 days. In each model, Sorafenib produces dose-dependent tumor growth inhibitionwith no evidence of toxicity, as measured by increased weight loss relative to control animals or drug-relatedlethality. In parallel to the antitumor efficacy studies, additional groups of four mice bearing 1

20、00 to 200 mg tumorsare treated orally with vehicle or Sorafenib (30 to 60 mg/kg), administered daily for 5 days, which is the shortesttreatment duration producing complete tumor stasis in the treated groups.Rats2In the study, 100- to 120-g male albino rats are utilized. After acclimatization period,

21、 rats are weighed and randomlydivided into three groups: Group 1 (normal control group; n=10) is given the vehicle daily for 8 weeks. Group 2(DENA group; n=15) receive i.p. single dose of 200 mg/kg DENA. Group 3 (Sorafenib group; n=12) is given Sorafeniborally at a dose of 10 mg/kg daily for 2 weeks

22、, 6 weeks after DENA i.p. injection. At the end of the experiment (8weeks), rats are weighed, anesthetized by ether, and killed, and their livers are dissected. Fresh liver is washed twicewith ice-cold saline, dried on clean paper towel, and weighed. Liver index is calculated as liver weight (g)/fin

23、al bodyweight (g)100. The liver is divided into five portions: one portion is preserved in 10 % formalin for histopathologicalexamination and the other portions are immediately frozen in liquid nitrogen and stored at 80C.MCE has not independently confirmed the accuracy of these methods. They are for

24、 reference only.戶使本產(chǎn)品發(fā)表的科研獻 Cancer Discov. 2019 Dec;9(12):1686-1695. Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. ACS Appl Mater Interfaces. 2017 Apr 12;9(14):12195-12202. Lab Chip. 2018 Nov 6;18(22):3379-3392. Cancer Lett. 2019 May 28;450:132-143.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Wilhelm SM, et al. BAY 43-9006 exhibits broad spectrum oral antitumor activity and targets the RAF/MEK/ERK pathway and re