PLX7904-PB04-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEPLX7904Cat.No.:HY-18997CASNo.:1393465-84-3Synonyms:PB04分⼦式:C₂₄H₂₂F₂N₆O₃S分⼦量:512.53作⽤靶点:Raf作⽤通路:MAPK/ERKPathway储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥30mg/mL(58.53mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM1.9511mL9.7555mL19.5111mL5mM0.3902mL1.9511mL3.9022mL10mM0.1951mL0.9756mL1.9511mL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。-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:≥2.5mg/mL(4.88mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性PLX7904有效的，选择性的BRAF抑制剂，在表达突变体RAS的细胞中，对BRAFV600E的IC50值约为5nM。IC50&TargetBRafV600E5nM(IC50,inmutantRASexpressingcells)体外研究PLX7904inhibitstheinvitrogrowthoftwomelanomacelllines(A375andCOLO829)andanadditionalhumancolorectalcancercelllineCOLO205thatexpressesBRAFV600EwithIC50valuesof0.17μM,0.53μM,and0.16μM,respectively,onaparwithvemurafenibIC50valuesinthesameassays(0.33μM,0.69μM,and0.25μM,respectively)[1].PLX7904andPLX8394potentlyinhibitERK1/2-drivenGAL4-Elk1reporteractivityinPRTcellsaswellasparentalcells.PLX7904andPLX8394treatmentat1μMconcentrationreducecolonyformationandviabilityinparentalcellstoasimilarlevelasPLX4720[2].PPLX7904potentlyinhibitsphosphorylationofERK1/2inmutantBRAFmelanomacellswithoutelicitingparadoxicalactivationinwild-typeBRAF,mutantNRASmelanomacells.PPLX7904inhibitsERK1/2inPLX470-resistantcelllines.PPLX7904treatmentpromotesapoptosisandinhibitsanchorage-independentgrowthofvemurafenibresistantcells[3].PROTOCOLCellAssay[2]ForMTTassays,2×103cellsareseededintriplicatein96wellsintheirregularculturemedium(containingPLX4720forPRTlines).Nextday,cellsarewashedtwicewithPBSandthenthemediumisreplenishedcontainingtheindicatedRAFinhibitor.Mediumischanged48hourslaterandafterafurther48hours,10μLof5mg/mLMTTreagentisaddedtowells,andincubatedforthreehours.Formazancrystalsarethensolubilizedovernightwitha1:10dilutionof0.1Mglycine(pH10.5)inDMSO.Wellsarethenanalyzedat450nMinaMultiskan®Spectrumspectrophotometer.ResultsdepictedarenormalizedtoDMSOconditionsandareacompositeofthreeindependentexperiments.Errorbarsshownarerepresentativeofthestandarderrorofmean(SEM).MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalCOLO205tumourcellsareculturedinDMEM10%FBS1%penicillin/streptomycinsupplementedwithbovineAdministration[1]insulin,at37°C.Balb/Cnudemice,female,6-8weeksold,weighingapproximately18-22g,areinoculatedsubcutaneouslyattherightflankwithCOLO205tumourcells(5×106)in0.1mLofPBSmixedwithmatrigel(50:50)fortumourdevelopment.Thetreatmentisstartedwhenmeantumoursizereachapproximately100mm3,witheightmiceineachtreatmentgrouprandomizedtobalancetheaverageweightandtumoursize.B9cellsareexpandedinDMEM10%FBS1%penicillin/streptomycin.Upontrypsinizationthecellsarewashedthreetimeswith20mLRPMI,andafterthefinalcentrifugationarere-suspended,counted,andadjustedbyvolumetoafinalconcentrationof5×107cellspermillilitre.B9xenograftsarestartedbyinjection2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEof5×106cellssubcutaneouslyin6-to7-week-oldfemalenudeBalb/cmice.Compounddosingstartswhentheaveragesizeoftumoursreach50-70mm3.Animalsareequallydistributedovertreatmentgroups(n=10)tobalancetheaveragetumoursizeandbodyweight.Animalsaredosedorallyfordays1-14twicedailyanddays15-28oncedailywithvehicle,vemurafenib50mgperkg,orPLX790450mgperkg.12-O-tetradecanoylphorbol-13-acetate(TPA)isputontheskinofallmicetwiceaweekduringweeks3and4atadoseof2µgin200µLacetone.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•NatCommun.2018Nov14;9(1):4775.•MolCellBiol.2016Sep26;36(20):2612-25.•ResearchSquarePrint.October27th,2022.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].ZhangC,etal.RAFinhibitorsthatevadeparadoxicalMAPKpathwayactivation.Nature.2015Oct22;526(7574):583-586.[2].BasileKJ,etal.InhibitionofmutantBRAFsplicevariantsignalingbynext-generation,selectiveRAFinhibitors.PigmentCellMelanomaRes.2014May;27(3):479-84[3].LeK,etal.SelectiveRAFinhibitorimpairsERK1/2phosphorylationandgrowthinmutantNRAS,vemurafenib-resistantmelanomacells