1、Product Data SheetEntinostatCat. No.: HY-12163CAS No.: 209783-80-2分式: CHNO分量: 376.41作靶點: HDAC; Autophagy; Apoptosis作通路: Cell Cycle/DNA Damage; Epigenetics; Autophagy; Apoptosis儲存式: Powder -20C 3 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 DMSO : 75 mg/mL (199.25 mM; Need ultrasonic)SolventMas

2、s1 mg 5 mg 10 mgConcentration制備儲備液1 mM 2.6567 mL 13.2834 mL 26.5668 mL5 mM 0.5313 mL 2.6567 mL 5.3134 mL10 mM 0.2657 mL 1.3283 mL 2.6567 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 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?shù)娜芙獍?。以下溶?/p>

3、案都請先按照 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，適當保存；體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當天使； 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: 2.08 mg/mL (5.53 mM); Clear solution此案可獲得 2.08 mg/mL (5.53 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例

4、，取 100 L 20.8 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: 2.08 mg/mL (5.53 mM); Clear solution此案可獲得 2.08 mg/mL (5.53 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 20.8 mg/mL 的澄 DMSO 儲備液加到 900 L 20% 的 SBE-C

5、D 理鹽溶液中，混合Page 1 of 2 www.MedChemE均勻。3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.08 mg/mL (5.53 mM); Clear solution此案可獲得 2.08 mg/mL (5.53 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 20.8 mg/mL 的澄 DMSO 儲備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 Entinostat 選擇性，可服的 HDAC class I 抑制劑，抑制 H

6、DAC1，HDAC2 和 HDAC3 的 IC50 分別為 243 nM，453nM 和 248 nM。IC & Target HDAC1 HDAC3 HDAC2243 nM (IC50) 248 nM (IC50) 453 nM (IC50)體外研究 Binding affinity of Entinostat (MS-275) against HDAC1 and HDAC2 is 282 nM and 156 nM, respectively1. Effects ofthe HDAC inhibitor Entinostat (MS-275) have been examined in h

7、uman leukemia and lymphoma cells (U937, HL-60,K562, and Jurkat) as well as in primary acute myelogenous leukemia blasts in relation to differentiation and apoptosis.MS-275 displays dose-dependent effects in each of the cell lines. When administered at a low concentration (e.g., 1 M), MS-275 exhibits

8、 potent antiproliferative activity, inducing p21CIP1/WAF1-mediated growth arrest and expressionof differentiation markers (CD11b) in U937 cells. Entinostat (MS-275) potently induces cell death, triggering apoptosisin 70% of cells at 48 h2.體內(nèi)研究 Entinostat (MS-27-275) at 49 mg/kg shows marked antitumo

9、r effects against KB-3-1, 4-1St, and St-4 tumor lines, anda moderate effect against Capan-1 tumor. Entinostat at 24.5 mg/kg and 12.3 mg/kg also shows significant effectsagainst these tumors. In addition, oral administration of Entinostat apparently increases the level of histoneacetylation in HT-29

10、tumor xenografts 4-24 h after the administration3. MS-275 administration (3.5 mg/kg i.p.) toExperimental autoimmune neuritis (EAN) rats once daily from the appearance of first neurological signs greatlyreduces the severity and duration of EAN and attenuated local accumulation of macrophages, T cells

11、 and B cells,anddemyelination of sciatic nerves. In addition, MS-275 treatment increases proportion of infiltrated Foxp3+ cells andanti-inflammatory M2 macrophages in sciatic nerves of EAN rats4.PROTOCOLKinase Assay 1 Biochemical assays of HDAC activity are carried out by Nanosyn in a reaction volum

12、e of 10 L in 384-wellmicroplates. A standard enzymatic reaction contains 5 L of 2 HDAC inhibitor (e.g., Entinostat), 4 L of 2.5 enzyme,and 1 L of 10 substrate in assay buffer (100 mM HEPES, pH 7.5, 25 mM KCl, 0.1% BSA, 0.01% Triton X-100, 1%DMSO). Final concentration of all HDACs in the enzymatic as

13、says is between 0.5 and 5 nM. A final substrateconcentration of 1 M FAM-RHKK(Ac)-NH2 or FAM-RHKK(trifluoroacetyl)-NH2 is used in all assays and found to bebelow the determined Km,app for each enzyme1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell

14、Assay 1 SH-SY5Y cells are maintained under normal culture conditions in a humidified incubator at 37C with 5% CO2 and aresplit twice weekly. Cells are plated in black 384-well plates at 2500 cells/well in 20-L volume of DMEM/F-12 culturemedia supplemented with 10% FBS and permitted to adhere overnig

15、ht. The following day, HDAC inhibitors (e.g.,Entinostat) are serially diluted in 100% DMSO, and this series is subsequently cross-diluted into culture media. 5 L ofcompound (e.g., Entinostat) diluted in media is added to the appropriate well of the cell plate to afford the indicatedfinal concentrati

16、on of inhibitor (e.g., Entinostat) with a final 0.1% DMSO. Treated cells are incubated under normalPage 2 of 3 www.MedChemEtissue culture conditions for 6, 24, 48, 72, or 96 h prior to quantitation of cellular ATP levels as measured usingCellTiter-Glo reagents. Similarly, after 6 h of incubation wit

17、h HDAC inhibitors (e.g., Entinostat), media from separatecell plates are aspirated, and cells are washed once with media containing no inhibitors. 25 L of mediasupplemented with 10% FBS and 0.1% DMSO (no inhibitors) is added back to the cells, and cellular ATP levels aredetermined using CellTiter-Gl

18、o after 24, 48, 72, or 96 h of incubation. Luminescence is measured at each time pointusing an Envision Instrument with a 0.1 s count time1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice3Administration 34 A2780 cells (9106) are suspended in

19、 PBS and are injected subcutaneously into the flank of nude mouse. For theother tumor lines, KB-3-1, HCT-15, 4-1St, Calu-3, St-4, Capan-1, and HT-29, tumors are passaged several times beforestarting in vivo antitumor testing, and a tumor lump (2-3 mm in diameter) is transplanted subcutaneously into

20、theflank of a nude mouse by using a trocar needle. Treatment (four or five mice in each experimental group) with thedrugs is started after the tumors are confirmed to have grown in the body (tumor size, 20-100 mm3). Entinostat isadministered orally once daily 5 days per week for 4 weeks. Tumor lengt

21、h and width are monitored twice weekly, andtumor volume is calculated.Rats4Male Lewis rats (8-10 weeks, 170-200 g) are housed under a 12-h light/dark cycle with free access to food and water.For therapeutic treatment, EAN rats receive i.p. injection of MS-275 (3.5 mg/kg) daily from day 10 to day 14

22、(sixrats/group). For injection, MS-275 is suspended in phosphate buffered saline (PBS) and the same volume (1 mL) ofPBS is given to control rats.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 Cell. 2019 Mar 7;176(6):1447-1460.e14. Proc Natl

23、 Acad Sci U S A. 2019 Feb 19;116(8):2961-2966. Clin Cancer Res. 2020 Jan 14. pii: clincanres.2884.2019. J Cell Physiol. 2019 May 23. Carcinogenesis. 2015 Feb;36(2):192-201.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Lauffer BE, et al. Histone deacetylase (HDAC) inhibitor kinetic rate constants correlate with cellular histone acetylation but not transcription and cellviability. J Biol Chem. 2013 Sep 13;288(37):26926-43.2. Rosato RR, et al. The histone deacetylase inhibitor MS-275 promotes d