Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEHI-TOPK-032Cat.No.:HY-101550CASNo.:487020-03-1分?式:C??H??N?OS分?量:369.4作?靶點(diǎn):TOPK作?通路:CellCycle/DNADamage儲(chǔ)存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:5mg/mL(13.54mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.7071mL13.5355mL27.0709mL5mM0.5414mL2.7071mL5.4142mL10mM0.2707mL1.3535mL2.7071mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲(chǔ)備液；?旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?，-20°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請(qǐng)先按照InVitro?式配制澄的儲(chǔ)備液，再依次添加助溶劑：(為保證實(shí)驗(yàn)結(jié)果的可靠性，澄的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)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(guò)程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過(guò)加熱和/或超聲的?式助溶)1.請(qǐng)依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥0.5mg/mL(1.35mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEBIOLOGICALACTIVITY?物活性HI-TOPK-032有效特異的TOPK抑制劑。體外研究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].體內(nèi)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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?OncolRep.2022Jul;48(1):125.2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESeemorecustomervalidationsonwww.MedChemEREFERENCES[1].KimDJ,etal.NovelTOPKinhibitorHI-TOPK-032effectivelysuppressescoloncancergrowth.CancerRes.2012Jun15;72(12):3060-8.McePdfHe