BIIB021-CNF2024-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEBIIB021Cat.No.:HY-10212CASNo.:848695-25-0Synonyms:CNF2024分?式:C??H??ClN?O分?量:318.76作?靶點:HSP;Autophagy作?通路:CellCycle/DNADamage;MetabolicEnzyme/Protease;Autophagy儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數據體外實驗DMSO:≥45mg/mL(141.17mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM3.1372mL15.6858mL31.3716mL5mM0.6274mL3.1372mL6.2743mL10mM0.3137mL1.5686mL3.1372mL請根據產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，避免反復凍融造成的產品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內使?，-20°C儲存時，請在1個?內使?。體內實驗請根據您的實驗動物和給藥?式選擇適當的溶解?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：(為保證實驗結果的可靠性，澄的儲備液可以根據儲存條件，適當保存；體內實驗的?作液，建議您現?現配，當天使?；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沉淀1/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE、析出現象，可以通過加熱和/或超聲的?式助溶)1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(7.84mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(7.84mM);ClearsolutionBIOLOGICALACTIVITY?物活性BIIB021(CNF2024)?種合成的具有?服活性的HSP90抑制劑，Ki值和EC50值分別為1.7nM和38nM。IC50&TargetHSP90HSP901.7nM(Ki)38nM(EC50)體外研究BIIB021bindsintheATP-bindingpocketofHsp90,interfereswithHsp90chaperonefunction,andresultsinclientproteindegradationandtumorgrowthinhibition.BIIB021inhibitstumorcell(BT474,MCF-7,N87,HT29,H1650,H1299,H69andH82)proliferationwithIC50from0.06-0.31μM.BIIB021inducesthedegradationofHsp90clientproteinsincludingHER-2,Akt,andRaf-1andup-regulatedexpressionoftheheatshockproteinsHsp70andHsp27[1].BIIB021inhibitsHodgkin'slymphomacells(KM-H2,L428,L540,L540cy,L591,L1236andDEV)withIC50from0.24-0.8μM.BIIB021showslowactivityinlymphocytesfromhealthyindividuals.BIIB021inhibitstheconstitutiveactivityofNF-κBdespitedefectiveIκB.BIIB021inducestheexpressionofligandsfortheactivatingNKcellreceptorNKG2DonHodgkin'slymphomacellsresultinginanincreasedsusceptibilitytoNKcell-mediatedkilling[2].BIIB021enhancestheinvitroradiosensitivityofHNSCCAcelllines(UM11BandJHU12)withacorrespondingreductionintheexpressionofkeyradioresponsiveproteins,increasesapoptoticcellsandenhancesG2arrest[3].BIIB021isconsiderablymoreactivethan17-AAGagainstadrenocorticalcarcinomaH295R.ThecytotoxicactivityofBIIB021isnotinfluencedbylossofNQO1orBcl-2overexpression,molecularlesionsthatdonotpreventclientlossbutarenonethelessassociatedwithreducedcellkillingby17-AAG.BIIB021isalsoactivein17-AAGresistantcelllines(NIH-H69,MESSADx5,NCI-ADR-RES,Nalm6)[4].體內研究OraladministrationofBIIB021leadstotumorgrowthinhibitioninmanytumorxenograftmodelsincludingN87,BT474,CWR22,U87,SKOV3andPanc-1[1].BIIB021effectivelyinhibitsgrowthofL540cytumoratadoseof120mg/kg[2].BIIB021significantlyenhancesantitumorgrowtheffectofradiationinJHU12xenograft[3].PROTOCOLKinaseAssay[1]Forfluorescencepolarizationcompetitionmeasurements,theFITC-geldanamycinprobe(20nM)isreducedwith2mMTCEPatroomtemperaturefor3hours,afterwhichthesolutionisaliquotedandstoredat-80°Cuntilused.RecombinanthumanHsp90α(0.8nM)andreducedFITC-geldanamycin(2nM)areincubatedina96-wellmicroplateatroomtemperaturefor3hoursinthepresenceofassaybuffercontaining20mMHEPES(pH7.4),50mMKCl,5mMMgCl2,20mMNa2MoO4,2mMDTT,0.1mg/mLBGG,and0.1%(v/v)CHAPS.Followingthispreincubation,BIIB021in100%DMSOisthenaddedtofinalconcentrationsof0.2nMto10μ2/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEM(finalvolume100μL,2%DMSO).Thereactionisincubatedfor16hoursatroomtemperatureandfluorescenceisthenmeasuredinanAnalystplatereader,excitation=485nm,emission=535nm.HighandlowcontrolscontainnoBIIB021ornoHsp90,respectively.Thedataarefittoafour-parametercurveandIC50isgenerated.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]AmodifiedtetrazoliumsaltassayisusedtomeasuretheIC50.Tumorcellsareaddedto96-wellplatesandpropagatedfor24hoursbeforeBIIB021addition.BIIB021isaddedtotheplatedcells.DMSO(0.03-0.003%)isincludedasavehiclecontrol.Afterincubationphenazinemethosulfate(stockconcentration1mg/mL)and3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,innersalt(stockconcentration2mg/mL)aremixedataratioof1:20andaddedtoeachwellofa96-wellplate.Reductionof3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,innersaltgivesrisetoasolubleformazanproductthatissecretedintotheculturemedium.After4hoursincubation,theformazanproductisquantitatedspectrophotometricallyatawavelengthof490nm.DataareacquiredusingSOFTmaxPROsoftware,and100%viabilityisdefinedastheA490ofDMSO-treatedcellsstainedwith3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,innersalt(themeanA490ofcellstreatedwithDMSOatarangeof0.03-0.003%).PercentviabilityofeachsampleiscalculatedfromtheA490valuesasfollows:%viability=(A490nmsample/A490nmDMSO-treatedcells×100).TheIC50isdefinedastheconcentrationthatgivesriseto50%inhibitionofcellviability.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalBALB/candathymicmiceareobtainedfromHarlanSprague-Dawleyatage6to8weeks.ThemiceareAdministration[1]maintainedinsterilizedcagesinaventilatedcagingsystemwitha12hlight/12hdarkphotoperiodattemperatureof21°Cto23°Candarelativehumidityof50±5%.Irradiatedpelletedfoodandautoclaveddeionizedwaterareprovidedadlibitum.Animalsareidentifiedbytheuseofindividuallynumberedeartags.N87tumorfragments(appr2mm3)herightflankoftheanimal.BIIB021isadministeredtoanimalsbearingN87stomachcarcinomatumorsatdosesof31,62.5,and125mg/kg,oncedaily,fromMondaytoFriday,for5weeks.Tumordimensionsaremeasuredusingcalipersandtumorvolumesarecalculatedusingtheequationforanellipsoidsphere(l×w2)/2=mm3,wherelandwrefertothelargerandsmallerdimensionscollectedateachmeasurement,respectively.Tumorvolumesaremeasuredandanimalsareweighedandmonitoredfortoxicityatleasttwiceweekly.Pvaluesarecalculatedusingthetwo-tailedStudent'sttesttoassessthedifferenceintumorvolumesbetweencontrolandtreatedgroups.P<0.05isconsideredsignificant.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產品發(fā)表的科研?獻?NatCommun.2017Sep4;8(1):422.?Viruses.2021,13(4),610.Seemorecustomervalidationsonwww.MedChemEREFERENCES3/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE[1].Lundgren,Karen.,etal.BIIB021,anorallyavailable,fullysyntheticsmall-moleculeinhibitoroftheheatshockproteinHsp90.Molecu