醉茄素A作用机制 - Medchemexpress - MCE中国

1、Product Data SheetWithaferin ACat. No.: HY-N2065CAS No.: 5119-48-2分式: CHO分量: 470.6作靶点: NF-B; Ferroptosis作通路: NF-B; Apoptosis储存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解性数据体外实验 DMSO : 100 mg/mL (212.49 mM; Need ultrasonic)SolventMass1 mg 5 mg 10 mgCon

2、centration制备储备液1 mM 2.1249 mL 10.6247 mL 21.2495 mL5 mM 0.4250 mL 2.1249 mL 4.2499 mL10 mM 0.2125 mL 1.0625 mL 2.1249 mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month (protect from light)。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。体内实验请根据您的实验动物和给药式选择适当的溶解案。以下

3、溶解案都请先按照 In Vitro 式配制澄清的储备液，再依次添加助溶剂：为保证实验结果的可靠性，澄 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 0.83 mg/mL (1.76 mM); Clear solution此案可获得 0.83 mg/mL (1.76 mM，饱和度未知) 的澄清溶液。以 1 mL 作液

4、为例，取 100 L 8.3 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 0.83 mg/mL (1.76 mM); Clear solution此案可获得 0.83 mg/mL (1.76 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 8.3 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-C

5、D 理盐溶液中，混合均匀。Page 1 of 2 www.MedChemE3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 0.83 mg/mL (1.76 mM); Clear solution此案可获得 0.83 mg/mL (1.76 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 8.3 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Withaferin A从南醉茄中分离到的甾体内酯，可以抑制 NF-kB 的活性，

6、靶作于波形蛋 (vimentin)，具有抗炎，抗肿瘤等功效。IC & Target NFB体外研究 Withaferin A has antiinflammatory activity, and potently inhibits NF-kB activation by preventing the TNF-inducedactivation of Ik-B kinase beta via a thioalkylation-sensitive redox mechanism1. Withaferin A also has anticanceractivity. Withaferin A tar

7、gets the IF protein vimentin, causes aggregation of vimentin filaments in bovine aorticendothelial cells (BAECs) at 3 M, and induces vimentin fragmentation in endothelial cells at 10 M2. Withaferin A(0.5, 1.5 M) alone or incombination with cisplatin (CIS) dose-dependently reduces tumorigenic potenti

8、al of ALDH1positive cancer stem cells (CSCs)3.体内研究 Withaferin A (2 mg/kg, i.p.) shows potent angiogenesis inhibitory activity via vimentin in mice2. Withaferin A (2mg/kg) combined with cisplatin (CIS) regulates the expression of ALDH1 marker, and downregulates the expression of securin in tumors col

9、lected from mice3.PROTOCOLCell Assay 3 Ovarian epithelial cancer cell line A2780 is maintained in RPMI1640 medium supplemented with insulin (5 g/mL),penicillin/streptomycin (100 IU/mL and 100 g/mL respectively) and 10% fetal bovine serum (FBS) from Hyclone.Withaferin A, cisplatin (CIS) and other rea

10、gents are prepared in DMSO. Cisplatin is prepared fresh each time3.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice2Administration 2 Vimentin homozygous-deficient mice (Vim/) and mice that are vimentin-heterozygous deficient (Vim+/) in the129

11、/Svev background are used in the assay. In brief, mice between 4 and 6 wk of age are anesthetized byintraperiteoneal (i.p.) injection of ketamine and xylazine. Corneas are topically anesthetized by application ofproparacain eye drop, and 1 L drop of dilute 0.15 M sodium hydroxide is applied for 1 mi

12、n. The cornea isimmediately washed extensively in saline solution, and corneal and limbal epithelium gently removed by scraping.The cornea is topically treated with atropine eye drop and covered with tobramycin and erythromycin antibiotic eyeointment. Withaferin A or 12-D WS (2 mg/kg solubilized in

13、DMSO) or vehicle (DMSO) is injected i.p. in respectivedrug or control groups of mice after their recovery from corneal injury, and subsequently every day thereafter for aperiod of 10 days. Mice are humanely killed and eyes enucleated. The anterior segment half of eyes are dissected andcorneal button

14、s are prepared. Corneal tissues are fixed in 100% acetone for 20 min, washed in PBS for 1 hr, andblocked for 18 hr in 1% BSA-PBS at 4C. Cornea whole-mount staining is performed by incubating tissues in FITC-conjugated rat anti-mouse CD31 antibody (1:333 dilution in 1% BSA-PBS) for 12 hr, washed away

15、 for 24 hr at 4C in1% BSA-PBS, and affixed to glass slides with a coverslip. Fluorescent staining is visualized on microscope, andquantified by importing digital images to NIH ImageJ2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedCh

16、emE户使本产品发表的科研献 Cancer Cell Int. 2020 May.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Kaileh M, et al. Withaferin a strongly elicits IkappaB kinase beta hyperphosphorylation concomitant with potent inhibition of its kinase activity. J BiolChem. 2007 Feb 16;282(7):

17、4253-64. Epub 2006 Dec 6.2. Bargagna-Mohan P, et al. The tumor inhibitor and antiangiogenic agent withaferin A targets the intermediate filament protein vimentin. Chem Biol. 2007Jun;14(6):623-34.3. Kakar SS, et al. Withaferin A (WFA) inhibits tumor growth and metastasis by targeting ovarian cancer stem cells. Oncota