PHA-680632-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEPHA-680632Cat.No.:HY-10178CASNo.:398493-79-3分⼦式:C₂₈H₃₅N₇O₂分⼦量:501.62作⽤靶点:AuroraKinase作⽤通路:CellCycle/DNADamage;Epigenetics储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥30mg/mL(59.81mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM1.9935mL9.9677mL19.9354mL5mM0.3987mL1.9935mL3.9871mL10mM0.1994mL0.9968mL1.9935mL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%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:≥2.08mg/mL(4.15mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.15mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.15mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性PHA-680632极光激酶抑制剂(aurora)，对auroraA，B和C的IC50值分别为27，135和120nM。IC50&TargetAuroraAAuroraBAuroraC27nM(IC50)135nM(IC50)120nM(IC50)体外研究PHA-680632shows30-to200-foldhigherIC50sofFLT3,LCK,PLK1,STLK2,VEGFR2,andVEGFR3comparedwithAuroraA.PHA-680632haspotentantiproliferativeactivityinawiderangeofcelltypes.TheIC50sare0.32,0.41,0.06,1.17,0.56,0.62,0.29,0.11,1.56,0.62,0.07,0.13,0.41μMforC33A,HeLa,HCT116,HT29,LOVO,A549,MCF7,A2780,U2OS,DU145,U937,HL60,NHDF.PHA-680632cancausepolyploidyintumorcells.PHA-680632celltreatmentinducesphenotypessimilartoAuroraAorBdepletion[1].PHA680632,inhibitscolonyformationindifferentcancercelllinesandinducedpolyploidy.Aurora-AinhibitionbyPHA680632enhancesradiationresponseincancercells,especiallyinp53-deficientcells[2].体内研究PHA-680632suppressestumorgrowthinanimalmodels.PHA-680632treatmentat45mg/kgdoseresultsin85%ofTGIwithoutsignsoftoxicityintheHL60humanacutemyelogenousleukemiaxenograftmodel.PHA-680632treatmentat60mg/kgi.v.b.i.d.for5daysresultsin78%ofTGIwithoutsignsoftoxicityintheA2780humanovariancarcinomamodel[1].PHA680632inassociationwithradiationleadstoanadditiveeffectincancercells,especiallyinthep53-deficientcells,butdoesnotactasaradiosensitiser[2].PROTOCOLKinaseAssay[1]InhibitionofkinaseactivitybyPHA-680632isassessedusingascintillationproximityassayformat.Inthisassay,thebiotinylatedsubstrateistransphosphorylatedbythekinaseinpresenceofATPtracedwithγ33-ATP.Thephosphorylatedsubstrateisthencapturedusingstreptavidin-coatedscintillationproximityassaybeadsandtheextentofphosphorylationisevaluatedbyβ-counteraftera4-hourrestforthefloatationofthebeadsonadense5MCsClsolution.InparticularapeptidederivedfromtheChocktidesequence(LRRWSLGL)isusedassubstrateforAuroraA,whereastheoptimizedpeptideAuroratide1isemployedforAuroraBandC.Theassayisruninarobotizedformaton96-wellplates.ThepotencyofthecompoundtowardAurorakinasesand29additionalkinasesbelongingtoourKinaseSelectivityScreeningpanelisevaluatedandtherelevantIC50saredetermined[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]Cellsareseededatdifferentdensitiesrangingfrom5,000to15,000cm2in24-wellplatewiththeappropriatecompletemedium.After24hours,platesaretreatedwithPHA-680632andincubatedfor72hoursat37°Cin2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE5%CO2atmosphere.Attheendofincubationtime,cellsaredetachedfromeachplateandcountedusingacellcounter.IC50sarecalculatedusingpercentageofgrowthversusuntreatedcontrol[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice:Tumourxenograftmicearerandomlyallocatedintofourgroups(sixmicepergroup):A,control;B,IRAdministration[2]alone,8ꢀGyin1day;C,PHA-680632alone,40ꢀmg/kg,b.i.d.,for4days;D,samedoseofPHA-680632combinedwithIR(24ꢀhafterthefirstadministrationofPHA680632,similarscheduleasIRalone)for4days.Drugorvehiclecontrol(samevolumeof20%Tween-80in5%glucosesolution)isadministeredintraperitoneally(i.p.).Thetumoursizeismeasuredtwiceaweekusinganelectroniccaliper.Follow-upofindividualmiceisconducted.Thetumourvolumeisestimatedfrom2Dtumourmeasurements[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].SonciniC,etal.PHA-680632,anovelAurorakinaseinhibitorwithpotentantitumoralactivity.ClinCancerRes.2006Jul1;12(13):4080-9.[2].TaoY,etal.EnhancementofradiationresponsebyinhibitionofAurora-AkinaseusingsiRNAoraselectiveAurorakinaseinhibitorPHA680632inp53-deficient