Apoptozole-Apoptosis-Activator-VII-DataSheet-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEApoptozoleCat. No.: HY-15098CAS No.: 1054543-47-3Synonyms: Apoptosis Activator VII分式: CHFNO分量: 625.56作靶點: HSP; Apoptosis作通路: Cell Cycle/DNA Damage; Metabolic Enzyme/Protease;Apoptosis儲存式: Powder -20C 3 years4C 2 yearsIn solvent

2、-80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 DMSO : 100 mg/mL (159.86 mM)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.00 mM); Clear solution1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE2. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (4.00 mM); Suspended solution;

3、Need ultrasonic3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (4.00 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Apoptozole是Hsc70 和 Hsp70 的 ATPase 結(jié)構(gòu)域抑制劑，可誘導(dǎo)凋亡，Kd 值分別為 0.21 和 0.14 M。IC50 & Target HSP70 HSC700.14 M (Kd) 0.21 M (Kd)體外研究 Apoptozole is an inhibitor of Hsc70 and Hsp70, which binds

4、 to Hsc70 and Hsp70, with Kds of 0.21 and 0.14 M, respectively. Apoptozole (1 M) induces apoptosis in P19 cells. Apoptozole shows inhibitory activitiesagainst several cancer cell lines, such as SK-OV-3 (ovarian cancer cells), HCT-15 (colon cancer cells), andA549 (lung cancer cells), with IC50s of 0.

5、22, 0.25, and 0.13 M, respectively 1. Apoptozole binds to theATPase domain of Hsc70 and Hsp70, but does not binds to other types of heat shock proteins such asHsp60, Hsp90 or Hsp40 2. Apoptozole (0-15 M) suppresses the growth of A549 cells, HeLa cells, andMDA-MB-231 cells, with IC50s ranging from 5

6、to 7 M. Apoptozole (5 or 10 M) shows no effect onassociations of HSP70 with ASK1, JNK, or BAX, and does not induce AIF-mediated caspase-independentapoptosis in HeLa cells 3.體內(nèi)研究 Apoptozole (4 mg/kg, i.p.) exhibits antitumor activities in nude mice xenografted with A549, RKO (colorectalcarcinoma), an

7、d HeLa cells 3.PROTOCOLKinase Assay 1 Stock solutions of malachite green (0.081% w/v), polyvinyl alcohol (2.3% w/v), and ammoniumheptamolybdate tetrahydrate (5.7% w/v in 6 M HCl) are prepared and stored at 4C. Three solutions aremixed with water in the ratio of 2 : 1 : 1 : 2 to prepare the malachite

8、 green reagent. For the determination ofthe ATPase activity of Hsc70, a master mixture of an ATPase domain of Hsc70 is prepared in assay buffer(100 mM Tris-HCl, 20 mM KCl, and 6 mM MgCl2, pH 7.4) as the final concentration of 1 mM. An aliquot (10mL) of this mixture is added into each well of a 96-we

9、ll plate. To this solution is added each compound(including Apoptozole) in assay buffer, and the plate is incubated for 30 min at room temperature. To start thereaction, 1 mL of 4 mM ATP is added to the solution. The final concentrations are 1 mM protein and 200 mMATP in 20 mL of assay buffer. After

10、 3 h incubation at 37C, 80 mL of the malachite green reagent is addedinto each well. The samples are mixed thoroughly and incubated at 37C for 15 min, and 10 mL of 34%sodium citrate is added to stop the nonenzymatic hydrolysis of ATP. The absorbance is determined at 620nm on a SpectraMax 340 PC 384

11、1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 3 Cells (5 105 per well) are plated in triplicate in 96-well plates in 0.1 mL of culture media with 10% FBS.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEAfter 24 hr, cells are treated with

12、 various concentrations of Apoptozole (0-15 M) in culture media with 3%FBS (final volume: 0.2 mL per well) for 18, 48, and 72 hr before treatment with MTT. Absorbance at 570 nmis measured using a UV microplate reader 3.MCE has not independently confirmed the accuracy of these methods. They are for r

13、eference only.Animal Male nude mice are housed in a pathogen-free room under controlled temperature and humidity. Mice agedAdministration 3 4 weeks are injected with tumor cells for the xenograft experiments. Viable A549 and RKO cells (5 106)and HeLa cells (5 106) are injected subcutaneously into th

14、e flank of mice. The A549 and RKO cellxenograft mice are immediately and randomly assigned to two groups. The first group (n = 10) is used as acontrol group and receives vehicle only. The second group (n = 10) receives intraperitoneal injections ofApoptozole (4 mg/kg/day) every other day for 2 weeks

15、. The HeLa cell xenograft mice are immediately andrandomly assigned to four groups. The first group (n = 10) is a control group receiving vehicle only. Thesecond group (n = 10) receives intraperitoneal injections of Apoptozole (4 mg/kg/day) every other day for 2weeks. The third group (n = 10) receiv

16、es intraperitoneal injections of doxorubicin (15 mg/kg/day) every otherday for 2 weeks. The fourth group (n = 10) receives intraperitoneal injections of Apoptozole (4 mg/kg/day)and doxorubicin (15 mg/kg/day) every other day for 2 weeks. Tumors in all mice are measured in twodimensions with calipers

17、every 3 days and tumor volumes are calculated using the formula volume = w l2/2, where w is the width at the widest point of the tumor and l is the length perpendicular to w. The resultsfrom individual mice are plotted as average tumor volumes versus time 3.MCE has not independently confirmed the ac

18、curacy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻 CNS Neurosci Ther. 2019 Jun 20.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Williams DR, et al. An apoptosis-inducing small molecule that binds to heat shock protein 70. Angew Chem Int Ed Engl.2008;47(39):7466-9.2. Cho HJ, et al. Probing the effect of an inhibitor of an ATPase domain of Hsc70 on clathrin-mediated endocytosis. Mol Biosyst. 2015Oct;11(10):2763-9.3. Ko SK, et al. A small molecule inhibitor of ATPase activity of HSP70