1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEWR-1065 dihydrochlorideCat. No.: HY-103640CAS No.: 14653-77-1分式: CHClNS分量: 207.16作靶點(diǎn): MDM-2/p53作通路: Apoptosis儲(chǔ)存式: Powder -20C 3 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) H2O : 100 mg/mL (482.72 mM)DMSO : 25 mg/mL (120.6

2、8 mM; Need ultrasonic)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 4.8272 mL 24.1359 mL 48.2719 mL5 mM 0.9654 mL 4.8272 mL 9.6544 mL10 mM 0.4827 mL 2.4136 mL 4.8272 mL請(qǐng)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(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期限。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?，配制前?qǐng)先配制澄清的儲(chǔ)備液，再依次

3、添加助溶劑(為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請(qǐng)依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 1.67 mg/mL (8.06 mM); Clear solution2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 1.67 mg/mL (8.06 mM); Clear solution3. 請(qǐng)依序添加每種溶劑： 1

4、0% DMSO 90% corn oil1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemESolubility: 1.67 mg/mL (8.06 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 WR-1065 dihydrochloride 可以保護(hù)正常組織免受某些癌癥藥物的毒性作，并通過(guò) JNK 依賴性信號(hào)通路激活p53。IC50 & Target p53 1體外研究 The DNA-binding activity is increased in a WR-1065 dihydrochloride (WR

5、-1065) concentration-dependentmanner. Cells treated with 1 mM WR-1065 dihydrochloride for 24 h reveal that all of the p53-induced genesanalyzed are transactivated following WR-1065 dihydrochloride treatment, in a p53-dependent manner.Significantly, treatment with WR-1065 dihydrochloride leads to a 3

6、-fold increase in luciferase expressiondriven by AP-1, and a 5-fold increase when this reporter gene is driven by NF-B, when these values arenormalized to the level of the cotransfected -galactosidase gene 2.體內(nèi)研究 The results show that wR-1065 dihydrochloride (WR-1065) attenuates the severity of 6-OH

7、DA-inducedcatalepsy (P 3.PROTOCOLKinase Assay 2 For Western analysis, cells are treated with 1 mM WR-1065 dihydrochloride (WR-1065) for 24 h, andsubconfluent cultures of cells are harvested and lysed in RIPA buffer supplemented with protease inhibitors.Protein concentrations are determined by a dete

8、rgent-compatible assay. Western blots are blocked andincubated in antibody in PBS/0.2% Tween 20/5% nonfat dry milk. Blots are incubated with 1 g/mL antibodyfor 1 h at room temperature, followed by washing in PBS/0.2% Tween 20 and incubation in peroxidase-conjugated secondary antibody and chemilumine

9、scence detection 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 2 To test the effects of paclitaxel in the presence or absence of WR-1065 dihydrochloride (WR-1065) on cellgrowth, cells are seeded in 96-well tissue culture dishes at 20% con

10、fluence and allowed to attach andrecover for at least 24 h. Varying combinations of paclitaxel alone or in combination with a 60 minpretreatment with 1 mM WR-1065 dihydrochloride are then added to each well, and the plates are incubatedfor an additional 48 h or 72 h. The number of surviving cells is

11、 determined by staining. The percentage ofcells killed by paclitaxel and/or WR-1065 dihydrochloride is calculated as the percentage decrease insulforhodamine B binding compare with control cells 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal S

12、eventy two rats are divided randomly into 9 equal groups: 1) Control group receives no injection and is leftAdministration 3 untreated for the entire period of the experiment as intact animals; 2) Sham operated group is subjected onlyto surgical procedure; 3) Vehicle (saline)-treated group receives

13、2 L saline (intra-SNc); 4) Lesioned groupreceives 6-hydroxydopamine; 5) Vehicle+6OHDA group receives saline as a vehicle 3 days once daily (2 2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEL/rat) before 6-OHDA injection; 6 to 8) Rats in these groups are pretreated with intra-SNc injection of WR-10

14、65 dihydrochloride (WR-1065) (20, 40 and 80 g/2 L/rat) 3 days before 6-OHDA injection; 9) Non-lesioned animals receive intra-SNc injection of WR-1065 dihydrochloride (80 g/2 L/rat) for three days 3.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFEREN

15、CES1. Pluquet O, et al. The cytoprotective aminothiol WR1065 activates p53 through a non-genotoxic signaling pathway involving c-Jun N-terminal kinase. J Biol Chem. 2003 Apr 4;278(14):11879-87.2. Shen H, et al. Binding of the aminothiol WR-1065 to transcription factors influences cellular response to anticancer drugs. J PharmacolExp Ther. 2001 Jun;297(3):1067-73.3. Afshin Kheradmand, et al. Effect of WR-1065 on 6-hydroxydopamine-induced catalepsy and IL-6 level in rats. Iran J Basic