G-5555-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEG-5555Cat.No.:HY-19635CASNo.:1648863-90-4分⼦式:C₂₅H₂₅ClN₆O₃分⼦量:492.96作⽤靶点:PAK作⽤通路:CellCycle/DNADamage;Cytoskeleton储存⽅式:4°C,sealedstorage,awayfrommoisture*Insolvent:-80°C,6months;-20°C,1month(sealed

storage,awayfrommoisture)溶解性数据体外实验DMSO:25mg/mL(50.71mM;ultrasonicandwarmingandheatto80°C)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.0286mL10.1428mL20.2856mL5mM0.4057mL2.0286mL4.0571mL10mM0.2029mL1.0143mL2.0286mL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(sealedstorage,awayfrommoisture)。-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:≥2.5mg/mL(5.07mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(5.07mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(5.07mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性G-5555有效PAK1抑制剂，对PAK1和PAK2的Ki值分别为3.7nM和11nM。IC50&TargetPAK1PAK23.7nM(Ki)11nM(Ki)体外研究G-5555isapotentPAK1inhibitorwithaKiof3.7nM.G-5555showsexcellentkinaseselectivityandinhibitsonlyeightoutofthe235kinasestestedotherthanPAK1withinhibition>70%:PAK2,PAK3,KHS1,Lck,MST3,MST4,SIK2,andYSK1.TheIC50sofG-5555againstSIK2,PAK2,KHS1,MST4,YSK1,MST3andLckare9,11,10,20,34,43,52nM,respectively.Ingeneral,G-5555demonstrateshighselectivityforthegroupIPAKs.ThereisnegligibleactivityforG-5555againstthehERGchannelwithIC50morethan10μMinapatchclampassay[1].G-5555potentlyinhibitsPAK2,withaKiof11nM.Inanarrayof23breastcancercelllines,G-5555hassignificantlygreatergrowthinhibitoryactivityincelllinesthatarePAK-amplifiedcomparedtonon-amplifiedlines[2].体内研究G-5555exhibitslowbloodclearanceandanacceptablehalf-life.Goodoralexposure(AUC=30μM•h)andhighoralbioavailability(F=80%)areachieved[1].InanH292non-smallcelllungercancer(NSCLC)xenograftstudyinmice,G-5555inhibitsphosphorylationofthePAK1/2downstreamsubstratemitogen-activatedproteinkinase1(MEK1)S298and,whenadministeredatanoraldoseof25mg/kgb.i.d.,imparts60%tumorgrowthinhibitioninthismodel13andaPAK1amplifiedbreastcancerxenograftmodel,MDAMB-175[2].PROTOCOLKinaseAssay[1]The10µLassaymixturescontain50mMHEPES(pH7.5),0.01%Brij-35,10mMMgCl2,1mMEGTA,2µMFRETpeptidesubstrate,andPAKenzyme(20pMPAK1;50pMPAK2;90pMPAK4).Incubationsarecarriedoutat22°Cinblackpolypropylene384-wellplates.Priortotheassay,enzyme,FRETpeptidesubstrateandseriallydilutedtestcompounds(G-5555,etc.)arepreincubatedtogetherinassaybuffer(7.5µL)for10minutes,andtheassayisinitiatedbytheadditionof2.5µLassaybuffercontaining4×ATP(160µMPAK1;480µMPAK2;16µMPAK4).Followingthe60-minuteincubation,theassaymixturesarequenchedbytheadditionofdevelopmentreagent,and1hourlatertheemissionsofCoumarin(445nm)andFluorescein(520nm)aredeterminedafterexcitationat400nm[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]Threemiceineachofthetwogroupsareadministered25mg/kgoralsuspensiondosetwice,withtheseconddosegiven6hoursafterthefirstdose.Thedosevolumesare5mL/kgfortheIVgroupand10mL/kgforthePOgroups.FollowingadministrationofG-5555,15µLofbloodiscollectedateachtimepointare2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEstoredat-70to-80°Cuntilanalysis[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•ActaPharmSinB.2020Apr;10(4):603-614.•Elife.2017Mar13;6:e22207.•EndocrRelatCancer.2019Aug;26(8):699-712.•ResearchSquarePreprint.2021Apr.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].NdubakuCO,etal.DesignofSelectivePAK1InhibitorG-5555:ImprovingPropertiesbyEmployinganUnorthodoxLow-pKaPolarMoiety.ACSMedChemLett.2015Oct31;6(12):1241-6.[2].RudolphJ,etal.ChemicallyDiverseGroupIp21-ActivatedKinase(PAK)InhibitorsImpartAcuteCardiovascularToxicitywithaNarrowTherapeuticWindow.JMedChem.