Amifostine-thiol-dihydrochloride-WR-1065-dihydrochloride-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEAmifostinethioldihydrochlorideCat.No.:HY-103640CASNo.:14653-77-1Synonyms:WR-1065dihydrochloride分⼦式:C₅H₁₆Cl₂N₂S分⼦量:207.16作⽤靶点:MDM-2/p53作⽤通路:Apoptosis储存⽅式:-20°C,sealedstorage,awayfrommoisture*Insolvent:-80°C,6months;-20°C,1month(sealed

storage,awayfrommoisture)溶解性数据体外实验H2O:≥100mg/mL(482.72mM)DMSO:25mg/mL(120.68mM;Needultrasonic)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM4.8272mL24.1359mL48.2719mL5mM0.9654mL4.8272mL9.6544mL10mM0.4827mL2.4136mL4.8272mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month(sealedstorage,awayfrommoisture)。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥1.67mg/mL(8.06mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥1.67mg/mL(8.06mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥1.67mg/mL(8.06mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Amifostinethiol(WR-1065)dihydrochloride可以保护正常组织免受某些癌症药物的毒性作⽤，并通过JNK依赖性信号通路激活p53。IC50&Targetp53[1]体外研究TheDNA-bindingactivityisincreasedinaAmifostinethioldihydrochloride(Amifostinethiol)concentration-dependentmanner.Cellstreatedwith1mMAmifostinethioldihydrochloridefor24hrevealthatallofthep53-inducedgenesanalyzedaretransactivatedfollowingAmifostinethioldihydrochloridetreatment,inap53-dependentmanner.Significantly,treatmentwithAmifostinethioldihydrochlorideleadstoa3-foldincreaseinluciferaseexpressiondrivenbyAP-1,anda5-foldincreasewhenthisreportergeneisdrivenbyNF-κB,whenthesevaluesarenormalizedtothelevelofthecotransfectedβ-galactosidasegene[2].体内研究TheresultsshowthatAmifostinethioldihydrochloride(Amifostinethiol)attenuatestheseverityof6-OHDA-inducedcatalepsy(P[3].PROTOCOLKinaseAssay[2]ForWesternanalysis,cellsaretreatedwith1mMWR-1065dihydrochloride(WR-1065)for24h,andsubconfluentculturesofcellsareharvestedandlysedinRIPAbuffersupplementedwithproteaseinhibitors.Proteinconcentrationsaredeterminedbyadetergent-compatibleassay.WesternblotsareblockedandincubatedinantibodyinPBS/0.2%Tween20/5%nonfatdrymilk.Blotsareincubatedwith1μg/mLantibodyfor1hatroomtemperature,followedbywashinginPBS/0.2%Tween20andincubationinperoxidase-conjugatedsecondaryantibodyandchemiluminescencedetection[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]TotesttheeffectsofpaclitaxelinthepresenceorabsenceofWR-1065dihydrochloride(WR-1065)oncellgrowth,cellsareseededin96-welltissueculturedishesat20%confluenceandallowedtoattachandrecoverforatleast24h.Varyingcombinationsofpaclitaxelaloneorincombinationwitha60minpretreatmentwith1mMWR-1065dihydrochloridearethenaddedtoeachwell,andtheplatesareincubatedforanadditional48hor72h.Thenumberofsurvivingcellsisdeterminedbystaining.Thepercentageofcellskilledbypaclitaxeland/orWR-1065dihydrochlorideiscalculatedasthepercentagedecreaseinsulforhodamineBbindingcomparewithcontrolcells[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEAnimalSeventytworatsaredividedrandomlyinto9equalgroups:1)ControlgroupreceivesnoinjectionandisleftAdministration[3]untreatedfortheentireperiodoftheexperimentasintactanimals;2)Shamoperatedgroupissubjectedonlytosurgicalprocedure;3)Vehicle(saline)-treatedgroupreceives2μLsaline(intra-SNc);4)Lesionedgroupreceives6-hydroxydopamine;5)Vehicle+6OHDAgroupreceivessalineasavehicle3daysoncedaily(2μL/rat)before6-OHDAinjection;6to8)Ratsinthesegroupsarepretreatedwithintra-SNcinjectionofWR-1065dihydrochloride(WR-1065)(20,40and80μg/2μL/rat)3daysbefore6-OHDAinjection;9)Non-lesionedanimalsreceiveintra-SNcinjectionofWR-1065dihydrochloride(80μg/2μL/rat)forthreedays[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•FrontCellDevBiol.2020Jul29;8:703.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].PluquetO,etal.ThecytoprotectiveaminothiolWR1065activatesp53throughanon-genotoxicsignalingpathwayinvolvingc-JunN-terminalkinase.JBiolChem.2003Apr4;278(14):11879-87.[2].ShenH,etal.BindingoftheaminothiolWR-1065totranscriptionfactorsinfluencescellularresponsetoanticancerdrugs.JPharmacolExpTher.2001Jun;297(3):1067-73.[3].AfshinKheradmand,etal.EffectofWR-1065on6-hydroxydopamine-inducedcatalepsyandIL-6level