高三尖杉酯碱作用机制 - Medchemexpress - MCE中国

1、Product Data SheetHomoharringtonineCat. No.: HY-14944CAS No.: 26833-87-4分式: CHNO分量: 545.62作靶点: STAT作通路: JAK/STAT Signaling; Stem Cell/Wnt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 : 50 mg/mL (91.64 mM)* means soluble, but saturation unk

2、nown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 1.8328 mL 9.1639 mL 18.3278 mL5 mM 0.3666 mL 1.8328 mL 3.6656 mL10 mM 0.1833 mL 0.9164 mL 1.8328 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

3、 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: 2.5 mg/mL (4.58 mM); Clear solution此案可获得 2.5 mg/mL

4、(4.58 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 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: 2.5 mg/mL (4.58 mM); Clear solution此案可获得 2.5 mg/mL (4.58 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL

5、的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合Page 1 of 2 www.MedChemE均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (4.58 mM); Clear solution此案可获得 2.5 mg/mL (4.58 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 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Homoharring

6、tonine (Omacetaxine mepesuccinate;HHT)起作。具有抗肿瘤特性的细胞毒性物碱，其通过抑制翻译延长来IC & Target STAT3体外研究 Homoharringtonine inhibits IL-6-induced STAT3 phosphorylation in a dose- and time-dependent manner.Homoharringtonine (HHT) inhibits cells growth, cell viability and colony formation, as well as induced cell apopt

7、osisthrough mitochondria pathway. The cytotoxicity of Homoharringtonine on human NSCLC cell lines is investigated,A549 (wild type EGFR) and NCI-H1975 (H1975, mutant EGFR with L858R and T790M), Gefitinib is used as a control. ByMTT assay, Homoharringtonine has moderate cytotoxicity to A549 with an IC

8、50 of 3.7 M and H1975 cells are moresensitive to Homoharringtonine with an IC50 of 0.7 M. Homoharringtonine inhibits the cell proliferation and growthof A549 cells and H1975 cells in a time- and dose-dependent manner through MTT assay. By trypan blue exclusionassay, Homoharringtonine rapidly reduces

9、 viable A549 and H1975 cells in a dose- and time-dependent manner.Homoharringtonine significantly inhibits the clonogenic ability of A549 and H1975 cells1.体内研究 Homoharringtonine (10 mg/kg) efficiently represses tumor growth compared to vehicle control or Gefitinib (P0.05).Additionally, Homoharringto

10、nine (HHT) treatment does not reduce the mice body weight, which suggests thatHomoharringtonine has no apparent side effect. All the mice are euthanized, the tumors are isolated and imaged andthe tumor sample cells are harvested to extract protein for determination if Homoharringtonine inhibits STAT

11、3phosphorylation via western blot. The level of STAT3 phosphorylation and MCL1 from Homoharringtonine treatmentgroup is significantly decreased compared to vehicle control or Gefitinib treatment. Meanwhile, consistant with theresults in the above, AKT1/2/3 and ERK1/2 phosphorylation is not inhibited

12、 with Homoharringtonine treatment. Tofurther examine the STAT3 phosphorylation in the xenograft tumor samples with different treatments, the tumorsamples are frozen and cutted into 10 m sections for fluorescent immunohistochemistry. Homoharringtoninetreatment inhibits STAT3 phosphorylation compared

13、to vehicle control or Gefitinib treatment1.PROTOCOLCell Assay 1 Human NSCLC cell lines MCF-10A, A549 and H1975 cells are seeded into 96-well plate and precultured for 24 h, thentreated with Homoharringtonine for 24 h or 48 h. Cell cytotoxicity is determined by MTT assay. The absorbance ismeasured at

14、 570 nm by Varioskan Flash Multimode Reader, and the cell death rate is calculated. Cell viability isestimated by trypan blue dye exclusion assay. The cells which exclude the dye are viable. Place 0.5 mL of a suitablecell suspension (dilute cells in complete medium without serum to 1106 cells per mL

15、) following adding 0.1 mL of0.4% trypan blue dye and mixing thoroughly, and then incubate at room temperature for 3 min and load into ahemacytometer to count cells in three separate fields (nonviable, deep blue cells as well as viable, clear cells). The cellviability rate is calculated. After staini

16、ng with Hoechst 33258 at 10 mg/mL for 10 min, cell death is observed by afluorescence microscope1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedChemEAnimal Mice1 Equal amounts of female and male nude immunodeficient mice (nu/nu), 6-

17、8 weeks old, are injectedAdministration 1 subcutaneously with NSCLC H1975 cells (2.5106) suspended in 100 L RPMI-1640 medium into the rightflank ofeach mouse. Treatments are started when the tumors reached a palpablesize. Mice are randomly divided into threegroups (n=10) and treated with Homoharring

18、tonine (10 mg/kg), Gefitinib (30 mg/kg) or vehicle control for 3 weeks.Vernier caliper measurements of the longest perpendicular tumor diameters are conducted along with the micetreatment to estimate the tumor volume, using the following formula: 4/3(width/2)2(length/2), representing the3-dimensiona

19、l volume of an ellipse tumor tissue. Animals are sacrificed when tumors reached to 2 cm or if the miceappeared moribund to prevent unnecessary morbidity to the mice. At the time of the animals death, tumors areexcised; cells are separatedand lyzed for western blot using anti-STAT3 antibody, anti-pSTAT3, anti-MCL1 and anti-GAPDH antibodies and immunohistochemistry.MCE has not independently confirmed the accuracy of these methods. They are for reference onl