Erlotinib-Hydrochloride-CP-358774-hydrochloride-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEErlotinibHydrochlorideCat.No.:HY-12008CASNo.:183319-69-9Synonyms:CP-358774hydrochloride;NSC718781hydrochloride;OSI-774hydrochloride分⼦式:C₂₂H₂₄ClN₃O₄分⼦量:429.9作⽤靶点:EGFR;Autophagy作⽤通路:JAK/STATSignaling;ProteinTyrosineKinase/RTK;Autophagy储存⽅式:4°C,sealedstorage,awayfrommoisture*Insolvent:-80°C,6months;-20°C,1month(sealed

storage,awayfrommoisture)溶解性数据体外实验DMSO:6.2mg/mL(14.42mM;Needultrasonicandwarming)H2O:<0.1mg/mL(insoluble)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.3261mL11.6306mL23.2612mL5mM0.4652mL2.3261mL4.6522mL10mM0.2326mL1.1631mL2.3261mL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)1/4MasterofBioactiveMolecules—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:≥0.5mg/mL(1.16mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥0.5mg/mL(1.16mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:0.5mg/mL(1.16mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY⽣物活性ErlotinibHydrochloride(CP-358774Hydrochloride)抑制EGFR激酶的IC50为2nM。IC50&TargetEGFR2nM(IC50)体外研究ErlotinibHydrochloride(CP-358774Hydrochloride)isalsoapotentinhibitoroftherecombinantintracellular(kinase)domainoftheEGFR,withanIC50of1nM.TheproliferationofDiFicellsisstronglyinhibitedbyErlotinibwithanIC50of100nMforan8-dayproliferationassay[1].ThecombinationofB-DIMandErlotinib(2μM)resultsinasignificantinhibitionofcolonyformationinBxPC-3cellswhencomparedwitheitheragentalone.ThecombinationofB-DIMandErlotinib(2μM)resultsinasignificantinductionofapoptosisonlyinBxPC-3cellswhencomparewiththeapoptoticeffectofeitheragentalone[2].体内研究Thereisa1.49-foldstatisticallysignificantdifferencebetweenAUC0-infafterp.o.administrationofErlotinib(5mg/kg)comparingBcrp1/Mdr1a/1b-/-andWTmice(7,419±1,720versus4,957±1,735ng*h/mLrespectively,P=0.01)[3].TheadministrationofErlotinib(10mg/kg/day,or20mg/kg/day)toBleomycin(BLM)-treatedratsshowsnoexacerbationoflunginjuriesinindicessuchasmacroscopicfindings,lungweights,histopathologicalscores(lunglesiondensityandlungfibrosisscore),andpulmonaryhydroxyproline(HyP)level.TheresultsuggeststhatErlotinibdoesnothaveanyexacerbatingeffectsonlunginjuriesinducedbyBLMinrats[4].PROTOCOLKinaseAssay[1]Thekinasereactionisperformedin50μLof50mMHEPES(pH7.3),containing125mMNaCl,24mMMgCl2,0.1mMNa3VO4,20μMATP,1.6μg/mLEGF,and15ngofEGFR,affinitypurifiedfromA431cellmembranes.ThecompoundinDMSOisaddedtogiveafinalDMSOconcentrationof2.5%.PhosphorylationisinitiatedbyadditionofATPandproceededfor8mmatroomtemperature,withconstantshaking.Thekinasereactionisterminatedbyaspirationofthereactionmixtureandiswashed4timeswithwashbuffer.PhosphorylatedPGTismeasuredby25mimofincubationwith50μLperwellHRP-conjugatedPY54antiphosphotyrosineantibody,dilutedto0.2μg/mLinblockingbuffer(3%BSAand0.05%Tween20inPBS).Antibodyisremovedbyaspiration,andtheplateiswashed4timeswithwashbuffer.ThecolonmetricsignalisdevelopedbyadditionofTMBMicrowellPeroxidaseSubstrate,50μLperwell,andstoppedbytheadditionof0.09Msulfuricacid,50μLperwell.Phosphotyrosineisestimatedbymeasurementofabsorbanceat450nm.Thesignalforcontrolsistypically0.6-1.2absorbanceunits,withessentiallynobackgroundinwells2/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEwithoutAlP,EGFR,orPOTandisproportionaltothetimeofincubationfor10mm[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]TotesttheviabilityofcellstreatedwithB-DIM,Erlotinib,orthecombination,BxPC-3andMIAPaCacellsareplated(3,000-5,000perwell)ina96-wellplateandincubatedovernightat37°C.ArangeofconcentrationsforbothB-DIM(10-50µM)andErlotinib(1-5µM)isinitiallytested.Basedontheinitialresults,theconcentrationofB-DIM(20µM)andErlotinib(2µM)arechosenforallassays.TheeffectsofB-DIM(20µM),Erlotinib(2µM),andthecombinationonBxPC-3andMIAPaCacellsaredeterminedbythestandardMTTassayafter72handisrepeatedthreetimes.ThecolorintensityismeasuredbyaTecanmicroplatefluorometerat595nm.DMSO-treatedcellsareconsideredtobetheuntreatedcontrolandassignedavalueof100%.Inadditiontotheaboveassay,wehavealsodoneclonogenicassayforassessingtheeffectsoftreatment[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[3]Administration[3][4]Bcrp1/Mdr1a/1b-/-andWTmicearetreatedp.o.ori.p.with5mg/kgErlotinib.Thei.p.administrationischosenassuminggooddrugabsorptionandcompletebioavailability.Samplingisdonefromthetipofthelateraltailveininthreeseries.Duringthefirstseries,wholebloodsamplesarecollectedat15minand0.5,1.5,5,and10hafterinjection.Basedontheresultsofthisinitialgroup,thesamplingtimesofthetwosubsequentseriesareadaptedto5and15minand0.5,1.5,4,and8hafterinjection.Aftercollection,thebloodsamplesareimmediatelycentrifugedandplasmaisstoredat-20°Cuntilhigh-performanceliquidchromatographicanalysistookplace.Rats[4]Seven-week-oldmaleCrl:CD(SD)rats(244-297g)areused.TheanimalsaretreatedwithErlotinibHydrochloride(10mg/kgand20mg/kg)orallybygavage.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CellRes.2020Oct;30(10):833-853.•NatImmunol.2022Feb;23(2):251-261.•JClinInvest.2020Mar2;130(3):1417-1430.•MolCell.2022May14;S1097-2765(22)00434-8.•SciTranslMed.2018Jul18;10(450).pii:eaaq1093.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].MoyerJD,etal.InductionofapoptosisandcellcyclearrestbyCP-358,774,aninhibitorofepidermalgrowthfactorreceptortyrosinekinase.CancerRes.1997Nov1;57(21):4838-48.[2].AliS,etal.Apoptosis-inducingeffectoferlotinibispotentiatedby3,3'-diindolylmethaneinvitroandinvivousinganorthotopicmodelofp