阿扎胞苷作用機制 - Medchemexpress - MCE中國

1、Product Data Sheet5-AzacytidineCat. No.: HY-10586CAS No.: 320-67-2分式: CHNO分量: 244.2作靶點: Nucleoside Antimetabolite/Analog; DNA Methyltransferase; Bacterial; Autophagy作通路: Cell Cycle/DNA Damage; Epigenetics; Anti-infection; Autophagy儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 mont

2、h溶解性數(shù)據(jù)體外實驗 DMSO : 31 mg/mL (126.95 mM)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制備儲備液1 mM 4.0950 mL 20.4750 mL 40.9500 mL5 mM 0.8190 mL 4.0950 mL 8.1900 mL10 mM 0.4095 mL 2.0475 mL 4.0950 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(fù)凍融造成的產(chǎn)品失效。儲備液的保存式和期限：-80C, 6

3、months; -20C, 1 month。-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(dāng)?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(dāng)保存；體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當(dāng)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

4、: 2.5 mg/mL (10.24 mM); Clear solution此案可獲得 2.5 mg/mL (10.24 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 (10.24 mM); Clear solutionPage 1 of 2 w

5、ww.MedChemE此案可獲得 2.5 mg/mL (10.24 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 (10.24 mM); Clear solution此案可獲得 2.5 mg/mL (10.24 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

6、DMSO 儲備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 5-Azacytidine (Azacitidine; 5-AzaC; Ladakamycin)是胞苷核苷類似物，特異型抑制 DNA 甲基化。5-Azacytidine 與DNA 結(jié)合共價捕獲 DNA甲噬 (autophagy)。轉(zhuǎn)移酶 (DNA methyltransferases)，有益于逆轉(zhuǎn)表觀遺傳變化。5-Azacytidine 誘導(dǎo)細(xì)胞IC & Target DNMT1 Nucleoside AutophagyAntimetabolite/Analog體外研究 Unmethylated C

7、pG islands associated with a variety of genes become partially or fully methylated in tumors and canbe reactivated by 5-Azacytidine1. 5-Azacytidine acts as weak inducers of erythroid differentiation of Frienderythroleukemia cells in the same concentration range where they affect DNA methyltransferas

8、e activity2. 5-Azacytidine inhibits L1210 cells with ID50 and ID90 values of 0.019 and circa 0.15 g/mL, respectively3.體內(nèi)研究 TdR-3H incorporation is significantly inhibited when the animals are exposed to 5-Azacitidine (100 mg/kg, i.p.) for 2 hror longer3.PROTOCOLKinase Assay 3 A crude cell-free extra

9、ct is isolated from LI 210 cells in culture by suspension of the cells in a given volume of0.05mol/LTris-HCl buffer, pH 7.4, and sonic extraction with a Biosonik at 70% maximal output for 30 sec. Thesupernatant is collected after centrifugation at 105,000 g for 60 min (4C) in a Model L Spinco ultrac

10、entrifuge. Thefinal protein concentration of the cell-free extracts is approximately 3 mg/mL. The extracts are used as the source ofenzymes. Ribonucleotide reductase activity is measured. A unit of enzyme is defined as the amount that catalyzeddCMP synthesis at a rate of 1 mmole/hr. The assay system

11、s for the measurement of pyrimidine nucleoside (CR) anddeoxynucleoside (TdR, CdR) kinases are essentially those described by Chu and Fischer. However, reactions areterminated by heating for 2 min in a boiling water bath, and the phosphorylated derivatives are isolated according tothe method of Bach.

12、 Fifty-jul aliquots are applied to 1-inch discs of diethylaminoethyl paper, which are then placed incounting vials and eluted with 0.5 mL of 0.5 mol/LPCA. After 1 hr, 12 mL of Diotol are added, and the radioactivity isdetermined.MCE has not independently confirmed the accuracy of these methods. They

13、 are for reference only.Cell Assay 3 Twenty mL of cells (circa 1104 cells/mL) are pipetted into sterilized culture tubes with screw caps and incubated at37C overnight. The experiment is initiated by the addition of 1 mL of 5-Azacytidine (5-azaCR) or medium for a givenperiod (from 0 to 240 min) prior

14、 to the addition of 1 mL of metabolite (or medium). Cell growth is determined twice aday for 3 days by means of a Model A Coulter counter. To determine IDSO and ID90 values, 5 mL of L1210 cells(5103 cells/mL) are incubated with the drug at 37C for 3 days, and cell growth is determined.MCE has not in

15、dependently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedChemEAnimal For the in vivo experiments, leukemic mice (bearing circa 1103 cells/animal) are given injections i.p. with 0.2 mL ofAdministration 3 5-Azacytidine (5-azaCR) of a given concentration. Two

16、hr later, the reaction is started by injecting 0.5 mL of labeledmetabolite (TdR-3H or UR-3H, 10 /Ci/12.5 g). After 1 hr, animals (3 mice/group) are killed by cervical fracture, andthe ascites are treated with heparin, collected, pooled, and then centrifuged immediately in a Sorvall refrigeratedcentr

17、ifuge Model R2C-B at 800g for 10 min (4C).MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) PLoS Pathog. 2020 Mar. Mol Oncol. 2018 Feb;12(2):180-195. Cell Death Dis. 2018 Apr 27;9(5):497. Cell Death Dis. 2018 Jan 26;9(2):129. Cell Commun Signa

18、l. 2019 Aug 14;17(1):94.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Christman JK. 5-Azacytidine and 5-aza-2-deoxycytidine as inhibitors of DNA methylation: mechanistic studies and their implications for cancer therapy.Oncogene. 2002 Aug 12;21(35):5483-95.2. Creusot F, et al. Inhibition of DNA methyltransferase and induction of Friend erythroleukemia cell differentiation by 5-azacytidineand 5-aza-2-de