Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEPHA-680632Cat.No.:HY-10178CASNo.:398493-79-3分?式:C??H??N?O?分?量:501.62作?靶點:AuroraKinase作?通路:CellCycle/DNADamage;Epigenetics儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:≥30mg/mL(59.81mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.9935mL9.9677mL19.9354mL5mM0.3987mL1.9935mL3.9871mL10mM0.1994mL0.9968mL1.9935mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；體內(nèi)實驗的?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?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶)1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:≥2.08mg/mL(4.15mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.15mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.15mM);ClearsolutionBIOLOGICALACTIVITY?物活性PHA-680632極光激酶抑制劑(aurora)，對auroraA，B和C的IC50值分別為27，135和120nM。IC50&TargetAuroraAAuroraBAuroraC27nM(IC50)135nM(IC50)120nM(IC50)體外研究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].體內(nèi)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?邊的抑制劑?師www.MedChemE5%CO2atmosphere.Attheendofincubationtime,cellsaredetachedfromeachplateandcountedusingacellcounter.IC50sarecalculatedusingpercentageofgrowthversusuntreatedcontrol[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice:Tumourxenograftmicearerandomlyallocatedintofourgroups(sixmicepergroup):A,control;B,IRAdministration[2]alone,8?Gyin1day;C,PHA-680632alone,40?mg/kg,b.i.d.,for4days;D,samedoseofPHA-680632combinedwithIR(24?hafterthefirstadministrationofPHA680632,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