Hotline:400-820-3792Inhibitors ? ScreeningLibraries ? Proteinswww.MedChemENN3201Cat.No.:HY-177578作用靶點: Antibody-DrugConjugates(ADCs);c-Kit;Apoptosis;Microtubule/Tubulin;ERK;Akt;Caspase作用通路: Antibody-drugConjugate/ADCRelated;ProteinTyrosineKinase/RTK;Apoptosis;CellCycle/DNADamage;Cytoskeleton;MAPK/ERKPathway;StemCell/Wnt;PI3K/Akt/mTOR儲存方式: PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY生物活性NN3201是一種靶向c-Kit的抗體-藥物偶聯(lián)物(ADC)，具有高親和力(KD=0.19pM)。NN3201由4-(3-Tosyl-2-(tosylmethyl)propanoyl)benzoicacid-glu(PEG24-Me)-val-cit-NH-benzyloxyformicacid-MMAE(HY-178219)和抗c-Kit人源單克隆抗體NN2101(HY-P991293)偶聯(lián)合成。NN3201能夠快速內(nèi)化并抑制SCF驅(qū)動的信號傳導，通過遞送有效載荷誘導細胞周期阻滯和凋亡(apoptosis)。由于FcγR結(jié)合力降低，NN3201不表現(xiàn)出Fc介導的ADCC和CDC效應(yīng)功能。NN3201在多種腫瘤模型中表現(xiàn)出顯著的c-Kit依賴性抗腫瘤功效。NN3201可用于小細胞肺癌(SCLC)、胃腸道間質(zhì)瘤(GIST)和急性髓系白血病(AML)的相關(guān)研究[1][2]。體外研究NN3201(1μg/mL,1h)exhibitsdose-dependentbindingtoc-Kit-highandc-Kit-mediumcelllines,whichsaturatedat315pM,withnobindingobservedtoc-Kit-lowornegativecelllines[1].NN3201(1μg/mL,0.5-24h)internalizesrapidly(within30minutes)andcontinuouslyinc-Kit-positiveGIST-T1cells,withthesignalincreasingforupto24hours,butitsinternalizationwasc-Kit-dependent[1].NN3201(3-7days)demonstratespotent,c-Kit-dependentcytotoxicitymediatedbyitsMMAEpayloadinapanelofpositivecelllines,withGI50valuesof0.09nM(GIST-T1),0.69nM(GIST-430),0.12nM(GIST-430/654),and17.06nM(NCI-H1048)[1].NN3201(0.01-1μg/mL,1h)dose-dependentlydecreasestheSCF-mediatedphosphorylationonc-Kit(Y719,Y568/570),Erk1/2,andAkt(S473)inNCI-H1048cells,whileonlypartiallysuppressingphosphorylation(pY568/570c-KitandpErk1/2)inGIST-430/654cells[1].NN3201(1μg/mL,1-3days)inducesc-Kitdegradationandsignalinginhibitionthroughcontinuousinternalization,whichtriggeredapoptosis,demonstratedbytheincreaseincleavedcaspase-3andcaspase-7[1].NN3201(5μg/mL,24-48h)inducescellcyclearrestintheG2/Mandsub-G1phases,whichisattributedtothemicrotubule-disruptingactionofitsreleasedMMAEpayload[1].NN3201(4μg/mL,7days)triggersapotentbystandereffect,mediatedbyfreeMMAEreleasedfrom1/5 MasterofBioactiveMolecules—您身邊的抑制劑大師www.MedChemENN3201-treatedGIST-430/654cells,whichkilledneighboringMCF7-GFPcancercellswithnegligiblec-Kitexpression[1].CellViabilityAssay[1]CellLine:MCF7-GFP+cellsConcentration:4μg/mLIncubationTime:7daysResult:Exhibitednegligiblecytotoxicity(GI50>20μg/mL)againstthec-Kit-negativeMCF7-GFPcellline,consistentwithitstarget-dependentmechanismandinsharpcontrasttothepotentactivityofitsfreepayload,MMAE(GI50=4.96nM).CellCycleAnalysis[1]CellLine:GIST-430/654andNCI-H1048cellsConcentration:1μg/mLIncubationTime:24and48hResult:InducedsubstantialcellcyclearrestattheG2/Mphaseandincreasedthesub-G1populationinbothcelllines.WesternBlotAnalysis[1]CellLine:GIST-430/654andMCI-H1048cellsConcentration:1μg/mLIncubationTime:1,2,and3daysResult:Inducedatime-dependentdegradationofc-Kitinbothcelllines.Increasedlevelsofcleavedcaspase-3andcaspase-7.Eventuallyinhibitedthec-KitdownstreamsignalingpathwaybysustainedinternalizationanddegradationoftheNN3201-c-Kitcomplexover3days.體內(nèi)研究NN3201(0.5-10mg/kg,i.v.,Q10Dx3orQ1Wx3)exhibitssignificantc-Kit-dependentanti-tumorefficaciesinvariousGISTandSCLCtumormicemodels[1].AnimalModel:FemaleC.B-17SCIDmice(5-6weeksold)subcutaneouslyinjectedwithGIST-T1cells[1]Dosage:0.5,1.5and3.0mg/kgAdministration:i.v.,Q10Dx32/5 MasterofBioactiveMolecules—您身邊的抑制劑大師www.MedChemEResult:Completelyregressedtumorvolumeforupto112daysat1.5and3mg/kg,withImatinib(HY-15463)(100mg/kg,p.o.,Q1Dx30)onlyinhibitedtumorprogressionduringthetreatmentperiod.Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:NOGmice(5-6weeksold)subcutaneouslyinjectedwithGIST-430/654cells[1]Dosage:1,3and5mg/kgAdministration:i.v.,Q1Wx3Result:Inducedsignificanttumorshrinkageat3mg/kgand5mg/kg,andstablycontrolleduntilday42and49,respectively.Inducedtumorgrowthinhibition(TGI)of47.2%onday21at1mg/kg,whichwassimilartothatby30mg/kgofSunitinib(HY-10255A)(30mg/kg,p.o.,Q1Dx21;TGIof45.2%onday21).Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:FemaleNOD/SCIDmicesubcutaneouslyimplantedwithGS5108tumors[1]Dosage:3,5and10mg/kgAdministration:i.v.,Q1Wx3Result:Induceddurabletumorstasisat10mg/kg,whichwasfollowedbysignificanttumorregressionforupto60days.Demonstratedsuperiorinvivoefficacyoverallstandard-of-care(SoC)treatmentsinthe3rdlineGISTmodels.Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:FemaleC.B-17SCIDmice(5-6weeksold)subcutaneouslyinjectedwithNCI-H526cells[1]Dosage:1,1.5,2,2.5and3mg/kgAdministration:i.v.,Q1Wx3Result:Inducedcompleteregressionofthetumorwithnoregrowthafter70and84daysat2.5and3mg/kg.Exhibited84.9%ofTGIcomparedtothevehiclecontrolonday17at2mg/kg.Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:FemaleC.B-17SCIDmice(5-6weeksold)subcutaneouslyinjectedwithNCI-H1048cells[1]3/5 MasterofBioactiveMolecules—您身邊的抑制劑大師www.MedChemEDosage:1,3,and5mg/kgAdministration:i.v.,Q1Wx3Result:ShowedTGIof95.1%onday21at1mg/kg,betterthanthecombinationofCarboplatin(HY-17393)(60mg/kg,i.p.,day0and10)andEtoposide(HY-13629)(3mg/kg.i.p.,day0~4and10~14;TGI=70.51%onday21).Inducedcompletetumorremissionforupto35daysand56daysat3mg/kgand5mg/kg,respectively.Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:FemaleC.B-17SCIDmice(5-6weeksold)subcutaneouslyinjectedwithNCI-H1048cells[1]Dosage:5mg/kgAdministration:i.v.,Q1Wx3Result:Wasabletoreducethetumorvolumetobelowbaseline,whereasneitherTopotecan(HY-13768)(0.83mg/kg,i.p.,BIWx3)norIrinotecan(HY-16562)(33mg/kg,i.v.,Q1Wx3)couldachievethis.Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:FemaleC.B-17SCIDmice(5-6weeksold)subcutaneouslyinjectedwithSHP-77cells[1]Dosage:1,3,and5mg/kgAdministration:i.v.,Q1Wx3Result:ExhibitednoinvivopotencyinSHP-77xenografts,wherec-Kitexpressionisnegligible,confirmingitsc-Kitdrivenproperty.Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:FemaleHsd:AthymicNude-Foxn1numicesubcutaneouslyimplantedwithCTG-1252tumors[1]Dosage:1,3,and5mg/kgAdministration:i.v.,Q1Wx3Result:Dose-dependentlyinhibitedtumorgrowth.Achieved70daysofcompleteregressionat5mg/kg.Inducednobodyweightlossbutratheranincreaseastumorsizeenlarges.AnimalModel:FemaleHsd:AthymicNude-Foxn1numicesubcutaneouslyimplantedwithCTG-20934/5 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