Dactolisib-BEZ235-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEDactolisibCat.No.:HY-50673CASNo.:915019-65-7Synonyms:BEZ235;NVP-BEZ235分⼦式:C₃₀H₂₃N₅O分⼦量:469.54作⽤靶点:PI3K;mTOR;Autophagy作⽤通路:PI3K/Akt/mTOR;Autophagy储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验5%TFA:8.33mg/mL(17.74mM;ultrasonicandwarmingandheatto60°C)DMSO:5mg/mL(10.65mM;ultrasonicandwarmingandheatto60°C)DMF:2mg/mL(4.26mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.1297mL10.6487mL21.2974mL5mM0.4259mL2.1297mL4.2595mL10mM0.2130mL1.0649mL2.1297mL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1/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE、析出现象，可以通过加热和/或超声的⽅式助溶)1.请依序添加每种溶剂：10%1-Methyl-2-pyrrolidinone>>90%PEG300Solubility:≥1.1mg/mL(2.34mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥0.52mg/mL(1.11mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥0.52mg/mL(1.11mM);Clearsolution4.请依序添加每种溶剂：50%PEG300>>50%salineSolubility:12.5mg/mL(26.62mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY⽣物活性Dactolisib(BEZ235)⼀种具有⼝服活性的、双重的pan-classIPI3K和mTOR抑制剂，作⽤于p110α/γ/δ/β和mTOR，IC50分别为4nM/5nM/7nM/75nM和20.7nM。Dactolisib(BEZ235)抑制mTORC1和mTORC2。IC50&Targetp110αp110βp110δp110γ4nM(IC50)75nM(IC50)7nM(IC50)5nM(IC50)p110α-H1047Rp110α-E545KmTORmTORC14.6nM(IC50)5.7nM(IC50)20.7nM(IC50)mTORC2Autophagy体外研究Dactolisib(BEZ235)potentlyinhibitsPI3KinanATPCompetitiveManner.Dactolisib(BEZ235)(250nM)significantlyreducedthephosphorylationlevelsofthemTORactivatedkinasep70S6K.Dactolisib(BEZ235)alsoleadstoareductionofS235/S236P-RPS6levelswithanIC50of6.5nM,suggestingthatDactolisib(BEZ235)candirectlyinhibitthemTORkinase,asthekinasedomainofmTORishighlyhomologoustotheoneofclassIAPI3K.TheactivityofDactolisib(BEZ235)againstmTORisconfirmedusingabiochemicalmTORK-LISAassay(IC50,20.7nM)[1].TheIC50sofDactolisib(BEZ235)forHCT116,DLD-1,andSW480celllinesare14.3±6.4,9.0±1.5,and12.0±1.6nM,respectively[2].体内研究Dactolisib(BEZ235)(45mg/kg,p.o.)treatmentinducescolonictumorregressioninaGEMmodelforsporadicPIK3CAwild-typeCRC[2].Dactolisib(BEZ235)(45mg/kg)isadministeredtoMENXrats(n=2eachgroup)byoralgavageandanimalsaresacrificed1or6hoursaftertreatment.ImmunostainsforP-AKTandP-S6showconsiderablereductionofthetwoproteins,andparticularlyofP-S6,6hoursafteradministrationofDactolisib(BEZ235)whencompareswithPEG-treatedrats.At6hoursaftertreatment,thepituitaryadenomasofDactolisib(BEZ235)-treatedratshasaproteomicprofilesignificantlydifferentfromthetumorsofplacebo-treatedrats[3].PROTOCOLCellAssay[2]HCT116(PIK3CAmutant;kinasedomainatH1047R),DLD-1(PIK3CAmutant;helicaldomainatE545K),2/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEandSW480(PIK3CAwild-type)humanCRCcelllines(ATCC)andisogenicDLD-1PIK3CAmutantandwild-typecellsaremaintainedinDMEM.Cellsareplatedatdifferentinitialdensities(HCT116:3,000cells/well,DLD-1:5,500cells/well,SW480:4,500cells/well,DLD-1PIK3CAmutant:7,000cells/well,andDLD-1PIK3CAwild-type:9,000cells/well)toaccountfordifferentialgrowthkinetics.After16hours,cellsareincubatedwithincreasingconcentrationsofBEZ235(10,100,1000nM),anddrug-containinggrowthmediumischangedevery24hours.Cellviabilityisassessed16hoursaftertheinitialplatingand48hoursafterinitiationofdrugtreatmentusingthecolorimetricMTSassayCellTiter96AQueousOneSolutionCellProliferationAssay.Cellviabilityafterdrugtreatmentisnormalizedtothatofuntreatedcellsalsogrownfor48hours.IC50valuesarecalculatedusing4parameternonlinearregressioninGraphPadPrism5[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[2]Administration[2][3]Tumor-bearingApcCKOmicearerandomlyassignedtotreatmentwitheithercontrolvehiclealone(n=8)or45mg/kgbodyweightBEZ235in10%1-methyl-2-pyrrolidone/90%PEG300(n=8)bydailyoralgavagefor28days.Thetreatmentdoseischosenbasedonliteratureindicatingthat40-50mg/kgbodyweightBEZ235effectivelytreatsmurinetumormodelswithoutadverseeffects.BaseonpharmacokineticstudiesdemonstratingmaximaltissueconcentrationonehourafterNVP-BEZ235administration,tumor-bearingmicearesacrificedonehourafterfinaltreatmentdose.Colonictumorvolumeisassessedusingcalipers(width×length×height)andtumorsareharvestedforbothwesternblotanalysisandimmunohistochemistry.Rats[3]MENX-affectedratsused.ThreedosesofBEZ235aretestedinMENXrats:20,30,and45mg/kg.Asthetwohigherdosescausesaweightloss>10%after10daysoftreatment,thedoseof20mg/kgisusedforfurtherstudies.ForMRIstudies,MENX-affectedratsat7to8monthsofage(withsizeableadenomasbutstillingoodgeneralhealth)aretreatedfor14dayswithBEZ235(20mg/kg)orplacebo(PEG)administereddailyperoralgavage.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•Nature.2018Aug;560(7719):499-503.•CellRes.2019Nov;29(11):895-910.•Blood.2019Oct17;134(16):1323-1336.•SciTranslMed.2018Jul18;10(450).pii:eaaq1093.•NatCommun.2017Jun8;8:15617.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].MairaSM,etal.IdentificationandcharacterizationofNVP-BEZ235,aneworallyavailabledualphosphatidylinositol3-kinase/mammaliantargetofrapamycininhibitorwithpotentinvivoantitumoractivity.