HI-TOPK-032-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEHI-TOPK-032Cat.No.:HY-101550CASNo.:487020-03-1分⼦式:C₂₀H₁₁N₅OS分⼦量:369.4作⽤靶点:TOPK作⽤通路:CellCycle/DNADamage储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:5mg/mL(13.54mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.7071mL13.5355mL27.0709mL5mM0.5414mL2.7071mL5.4142mL10mM0.2707mL1.3535mL2.7071mL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、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:≥0.5mg/mL(1.35mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEBIOLOGICALACTIVITY⽣物活性HI-TOPK-032有效特异的TOPK抑制剂。体外研究HI-TOPK-032stronglysuppressesTOPKkinaseactivitybuthaslittleeffectonextracellularsignal-regulatedkinase1(ERK1),c-jun-NH2-kinase1,orp38kinaseactivities.HI-TOPK-032occupiestheATP-bindingsiteofTOPKandfitsthebindingsiteverywell.ThecompoundformshydrogenbondswithGLY83andASP151andhasahydrophobicinteractionwithLYS30.However,HI-TOPK-032atthehighestconcentration(5μM)alsoinhibitsMEK1activityby40%.HI-TOPK-032alsoinhibitsanchorage-dependentand-independentcoloncancercellgrowthbyreducingERK-RSKphosphorylationaswellasincreasingcoloncancercellapoptosisthroughregulationoftheabundanceofp53,cleavedcaspase-7,andcleavedPARP[1].体内研究Treatmentofmicewith1or10mg/kgofHI-TOPK-032significantlyinhibitsHCT-116tumorgrowthbymorethan60%relativetothevehicle-treatedgroup.MicearewelltoleratedwithHI-TOPK-032treatment.Theexpressionofp53isstronglyinduced,andphosphorylationofERKandRSK,adirectdownstreamproteinofERK,ismarkedlyinhibitedintheHI-TOPK-032-treatedgroup[1].PROTOCOLKinaseAssay[1]TheeffectofHI-TOPK-032onERK1,JNK1,andp38activityisassessedbyaninvitrokinaseassayusingERK1(active,500ng),inactiveRSK2(ERK1substrate,1μg),JNK1(active,50ng),c-Jun(JNK1substrate,1μg)andp38(active,200ng),andATF2(p38substrate,500ng)with[γ-32P]ATP.Briefly,thereactioniscarriedoutinthepresenceof10μCiof[γ-32P]ATPwithHI-TOPK-032(0.5,1,2,5μM)in40μLofreactionbuffer.Afterincubationatroomtemperaturefor30minutes,thereactionisstoppedbyadding10μLproteinloadingbufferandthemixtureisseparatedbySDS[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]HCT-116coloncancercellsaretreatedwithdifferentdosesofHI-TOPK-032(1,2,5μM).Afterincubationfor1,2,or3days,20μLofCellTiter96AQueousOneSolutionisaddedandthencellsareincubatedfor1hourat37°Cina5%CO2incubator.Absorbanceismeasuredat492nm[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice:Micearedividedinto4groups:(i)untreatedvehiclegroup;(ii)1mgHI-TOPK-032/kgofbodyweight;Administration[1](iii)10mgHI-TOPK-032/kgofbodyweight;and(iv)nocellsand10mgHI-TOPK-032/kgofbodyweight.HCT-116cellsaresuspendedinserum-freeMcCoy5Aotherightflankofeachmouse.HI-TOPK-032orvehicleisinjected3timesperweekfor25days.Tumorvolumeiscalculated[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•OncolRep.2022Jul;48(1):125.2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESeemorecustomervalidationsonwww.MedChemEREFERENCES[1].KimDJ,etal.NovelTOPKinhibitorHI-TOPK-032effectivelysuppressescoloncancergrowth.CancerRes.2012Jun15;72(12):3060-8.McePdfHe