Amifostine-thiol-dihydrochloride-WR-1065-dihydrochloride-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEAmifostinethioldihydrochlorideCat.No.:HY-103640CASNo.:14653-77-1Synonyms:WR-1065dihydrochloride分?式:C?H??Cl?N?S分?量:207.16作?靶點(diǎn):MDM-2/p53作?通路:Apoptosis儲存?式:-20°C,sealedstorage,awayfrommoisture*Insolvent:-80°C,6months;-20°C,1month(sealed

storage,awayfrommoisture)溶解性數(shù)據(jù)體外實(shí)驗(yàn)H2O:≥100mg/mL(482.72mM)DMSO:25mg/mL(120.68mM;Needultrasonic)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM4.8272mL24.1359mL48.2719mL5mM0.9654mL4.8272mL9.6544mL10mM0.4827mL2.4136mL4.8272mL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：-80°C,6months;-20°C,1month(sealedstorage,awayfrommoisture)。-80°C儲存時(shí)，請?jiān)?個(gè)?內(nèi)使?，-20°C儲存時(shí)，請?jiān)?個(gè)?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)動物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實(shí)驗(yàn)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(shí)驗(yàn)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)1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥1.67mg/mL(8.06mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥1.67mg/mL(8.06mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥1.67mg/mL(8.06mM);ClearsolutionBIOLOGICALACTIVITY?物活性Amifostinethiol(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JNK依賴性信號通路激活p53。IC50&Targetp53[1]體外研究TheDNA-bindingactivityisincreasedinaAmifostinethioldihydrochloride(Amifostinethiol)concentration-dependentmanner.Cellstreatedwith1mMAmifostinethioldihydrochloridefor24hrevealthatallofthep53-inducedgenesanalyzedaretransactivatedfollowingAmifostinethioldihydrochloridetreatment,inap53-dependentmanner.Significantly,treatmentwithAmifostinethioldihydrochlorideleadstoa3-foldincreaseinluciferaseexpressiondrivenbyAP-1,anda5-foldincreasewhenthisreportergeneisdrivenbyNF-κB,whenthesevaluesarenormalizedtothelevelofthecotransfectedβ-galactosidasegene[2].體內(nèi)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?邊的抑制劑?師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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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