依维莫司作用机制 - Medchemexpress - MCE中国

1、Product Data SheetEverolimusCat. No.: HY-10218CAS No.: 159351-69-6分式: CHNO分量: 958.22作靶点: mTOR; FKBP; Autophagy; Apoptosis作通路: PI3K/Akt/mTOR; Apoptosis; Autophagy; Immunology/Inflammation储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 54 mg/mL (56.35 mM)H2O : 0.

2、1 mg/mL (insoluble)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 1.0436 mL 5.2180 mL 10.4360 mL5 mM 0.2087 mL 1.0436 mL 2.0872 mL10 mM 0.1044 mL 0.5218 mL 1.0436 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 mont

3、h。-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 (2.61 mM

4、); Clear solution此案可获得 2.5 mg/mL (2.61 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 (2.61 mM); Precipitated solutionPage 1 of 2 www.MedChemE此案可获得

5、 2.5 mg/mL (2.61 mM，饱和度未知)以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (2.61 mM); Clear solution此案可获得 2.5 mg/mL (2.61 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。B

6、IOLOGICAL ACTIVITY物活性 Everolimus (RAD001)种 Rapamycin 的衍物，也 种有效的，选择性的和服活性的 mTOR1 抑制剂。Everolimus 与 FKBP-12 结合可产免疫抑制复合物。Everolimus 抑制肿瘤细胞增殖并诱导细胞凋亡 (apoptosis) 和噬 (autophagy)。Everolimus 具有有效的免疫抑制和抗癌活性。IC & Target mTOR5-6 nM (IC50)体外研究 Everolimus (RAD001) is an orally active derivative of rapamycin that

7、inhibits the Ser/Thr kinase, mTOR1. In both thesensitive murine B16/BL6 melanoma (IC50, 0.7 nM) and the insensitive human cervical KB-31 (IC50, 1,778 nM),antiproliferative concentrations of Everolimus results in total dephosphorylation of S6K1 and the substrate S6 and ashift in the mobility of 4E-BP

8、1, which is indicative of a reduced phosphorylation status3. Everolimus exhibits a dose-dependent inhibition in both the total cells and the stem cells from the BT474 cell line and the primary breast cancercells, albeit with different degrees of growth inhibition. Compare with the total cells, Evero

9、limus is less effective ingrowth inhibition in the stem cells at all tested concentrations (P0.001). The IC50 values of Everolimus for BT474 andthe primary CSCs are 2,054 and 3,227 nM, or 29 times and 21 times greater than the IC50 values for theircorresponding total cells, respectively4.体内研究 Everol

10、imus is orally active in both mice and rats, producing an antitumor effect that is characterized by dramaticreduction in tumor growth rates as opposed to producing tumor regressions. In the rat CA20498 model, daily treatment with Everolimus (0.5 or 2.5 mg/kg) dose-dependently inhibits growth, and in

11、termittent dosing using ahigher dose of 5 mg/kg (once or twice per week) also shows similar antitumor efficacy. Inhibition by Everolimus ischaracterized by sustained suppression rather than regression and is not associated with any body weight loss1. Theeffect of Everolimus treatment (0.1-10 mg/kg/d

12、) is selective and differ from the effects of PTK/ZK (100 mg/kg). Witheither growth factor, Everolimus dose-dependently increases the hemoglobin content (convert to blood equivalentsand indicative of the number of vessels as well as vascular leakiness) but reduces the Tie-2 content (number ofendothe

13、lial cells indicative of the number of vessels) and this is significant for VEGF stimulation but not bFGFstimulation. The pharmacokinetics of Everolimus in mice shows that maximum levels of only 0.1 M are achieved in ahuman tumor xenograft following a single administration, whereas plasma levels rea

14、ch 1 to 3 M for 4 h3.PROTOCOLCell Assay 2 Tumor cells are plated into 96-well plates at densities ranging from 500 to 5,000/100 L/well, with repeatexperiments being done at an optimal cell number, typically 1,000 to 2,000 per well, and incubated overnight. Cellsare exposed to Everolimus and incubate

15、d for 4 days and the cell number is determined by methylene blue staining.For this, 50 L glutaraldehyde 20% (v/v) is added to the wells incubated for 10 min at room temperature. Theculture medium is aspirated, cells are washed with distilled water, and 100 L methylene blue 0.05% (w/v) in water isadd

16、ed and incubated for 10 min at 37C. Stained cells are washed three times with water, 200 L HCl 3% (v/v) isPage 2 of 3 www.MedChemEadded, and the plate shaken at room temperature for 20 min. The absorbance of each well is determined at 650 nm.The IC50 values are calculated using Softmax 2.0 software2

17、.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice2Administration 2 Everolimus, PTK/ZK, and their respective vehicles are prepared each day just before administration to animals and theadministration volume is individually adjusted based on an

18、imal body weight. In C57/BL6 mice, Everolimus isadministered at doses ranging from 0.1 to 10 mg/kg/d orally (10 mL/kg) and predominantly at 2.5 to 10 mg/kgbecause these doses provides the maximum effect. PTK/ZK is administered at 50 to 100 mg/kg/d orally.Rats2Wistar-Furth rats are divided into two e

19、qual groups based on body weight and treated either with vehicle orEverolimus (10 mg/kg/d orally in mice and 5 mg/kg three times per week orally in rats). Directly after the firstmeasurement at baseline (day 0), Everolimus or vehicle is administered orally by gavage (10 mL/kg) for up to 7 daysmaximu

20、m with subsequent magnetic resonance measurements made within 30 min of the last dose.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Nature. 2016 Dec 1;540(7631):119-123. Nat Med. 2016 Jul;22(7):723-6. Sci Transl Med. 2013 Jul 31;5(196):196

21、ra99. J Clin Invest. 2017 Sep 1;127(9):3339-3352. Cell Syst. 2020 Jan 22;10(1):66-81.e11.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. OReilly T, et al. Biomarker Development for the Clinical Activity of the mTOR Inhibitor Everolimus (RAD001): Processes,Limitations, and FurtherProposals. Transl Oncol. 2010 Apr;3(2):65-79.2. Lane HA, et al. mTOR inhibitor RAD00