AM580-CD336-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEAM580Cat. No.: HY-10475CAS No.: 102121-60-8Synonyms: CD336; NSC608001; Ro 40-6055分式: CHNO分量: 351.44作靶點(diǎn): RAR/RXR; Autophagy作通路: Metabolic Enzyme/Protease; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1

2、month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 45 mg/mL (128.04 mM)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (7.11 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% corn oil1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemESolubility: 2.5 mg/mL (7.11 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 AM580是選擇性的 RAR 激動(dòng)劑

3、，IC50 和 EC50 分別為8 nM，0.36 nM。體外研究 In the presence of G-CSF, AM580 (at 10-8 M) produces a remarkable induction in LAP mRNA of NB4 cells.At a concentration of 10-5 M, AM580 and ATRA, in combination with G-CSF, induce almost the same level ofLAP transcript. AM580 (at 10-8 M) leads to an approximately s

4、ixfold increase in the steady-state levels of thetranscript coding for the G-CSF receptor in NB4 cells 1. AM580 (50 nM) increases caspase-3 expression inall of the colonies, and in 30% of the colonies induce acinar-like cavitation 2. Knockdown of RAR1 inprimary Myc cells using shRAR1 followed by Am5

5、80 treatment results in an even higher level of CRBP1expression, showing that in these cells RAR has a repressive effect on the RAR target gene CRBP1.Am580 (200 nM) enhances the anti-proliferative effect exhibited by RAR knockdown in the MCF-10A andMCF-7 cell lines but not in the MDA-MB-231 cells 3.

6、體內(nèi)研究 Am580 (0.3 mg/kg/day) treatment has a more profound effect on tumor-free survival of MMTV-wnt1 mice, theeffect being noticeable even in early appearing tumors, and no overt toxicity is found in liver, lungs, kidney,and spleen. Am580 treatment reduces substantially and equally the level of hyper

7、plasia in both transgenicglands 2. Treatment of MMTV-Myc mice with the RAR-selective agonist Am580 leads to significantinhibition of mammary tumor growth, lung metastasis and extends tumor latency in 63% of mice 3.PROTOCOLKinase Assay 1 Approximately 1106 NB4, HL-60, and APL fresh leukemic cells or

8、CML neutrophils are harvested, pellettedby centrifugation at 400 g for 10 minutes, washed once with 0.9% NaCl, and centrifuged again. The washedcell pellet is resuspended in homogenization buffer (1 mM MgCl2, 1 mM CaCl2, 20 mM ZnCl2, 0.1 mM NaCl,0.1% vol/vol Triton X-100, 50 mM Tris/HCl, pH 7.4) and

9、 disrupted by vigorous pipetting. The homogenate isused for the LAP assay, which is performed with p-nitrophenol phosphate as substrate according to theinstructions of the manufacturer. LAP activity is normalized for the content of protein in the sample. Proteinsare measured according to the Bradfor

10、d method using BSA fraction V as a standard. One unit of LAP activityis defined as the amount of enzyme capable of transforming 1 nmol of substrate in 1 minute at 25C. Enzymeassays are performed in conditions of linearity relative to the substrate and to the concentration of proteins.MCE has not ind

11、ependently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 MCF-10A (2104) control cells or overexpressing RAR are seeded in triplicates in 6-well culture dishes.After 24 hrs, cells are washed with PBS, incubated in 2 mL of DMEM-F12 culture medium, detached andcounte

12、d every 24 hrs. Statistical significance is determined by t-test. pRB and p27 expression is tested byimmunofluorescence analysis of control MCF-10A monolayers and monolayers stably transfected with pSG5-RAR expression vector, using the same antibodies described for the IHC analysis.MCE has not indep

13、endently confirmed the accuracy of these methods. They are for reference only.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEAnimal Four months old uniparous (1 pregnancy/lactation cycle) MMTV-neu and 6 weeks old nulliparous MMTV-Administration 1 wnt1 female mice (50 mice/group) are treated with t

14、he RAR agonist AM580 (0.3 mg/kg body weight permouse per day) in the diet (Purina 5053) by adding 1.5 mg AM580 per kg of Purina 5053 diet. Mice thatdevelop tumors within the first month of treatment are removed from the study. Mice are palpated twice aweek and tumor appearance is recorded. Once palp

15、able, the size of the tumors is measured weekly. Tumor-free survival is calculated from Kaplan-Meier curves and statistical significance is determined by the Log-ranktest for the survival studies and t-test for the tumor growth studies. Metastasis is evaluated by removing thelungs of all the anesthe

16、tized mice, selecting randomLy 20 mice per group and inspecting the lung surface forlesions using a stereoscope.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Giann M, et al. AM580, a stable benzoic derivative of retinoic acid, has powerfu

17、l and selective cyto-differentiating effects on acutepromyelocytic leukemia cells. Blood. 1996 Feb 15;87(4):1520-31.2. Lu Y, et al. Mechanism of inhibition of MMTV-neu and MMTV-wnt1 induced mammary oncogenesis by RARalpha agonist AM580.Oncogene. 2010 Jun 24;29(25):3665-76.3. Bosch A, et al. Reversal by RAR agonist Am580 of c-Myc-induced imbalance in RAR/RAR expression during MMTV-