BIIB021-CNF2024-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEBIIB021Cat.No.:HY-10212CASNo.:848695-25-0Synonyms:CNF2024分⼦式:C₁₄H₁₅ClN₆O分⼦量:318.76作⽤靶点:HSP;Autophagy作⽤通路:CellCycle/DNADamage;MetabolicEnzyme/Protease;Autophagy储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥45mg/mL(141.17mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM3.1372mL15.6858mL31.3716mL5mM0.6274mL3.1372mL6.2743mL10mM0.3137mL1.5686mL3.1372mL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%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(7.84mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(7.84mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性BIIB021(CNF2024)⼀种合成的具有⼝服活性的HSP90抑制剂，Ki值和EC50值分别为1.7nM和38nM。IC50&TargetHSP90HSP901.7nM(Ki)38nM(EC50)体外研究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].体内研究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—您⾝边的抑制剂⼤师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.户使⽤本产品发表的科研⽂献•NatCommun.2017Sep4;8(1):422.•Viruses.2021,13(4),610.Seemorecustomervalidationsonwww.MedChemEREFERENCES3/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE[1].Lundgren,Karen.,etal.BIIB021,anorallyavailable,fullysyntheticsmall-moleculeinhibitoroftheheatshockproteinHsp90.Molecu