G-5555-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEG-5555Cat.No.:HY-19635CASNo.:1648863-90-4分?式:C??H??ClN?O?分?量:492.96作?靶點(diǎn):PAK作?通路:CellCycle/DNADamage;Cytoskeleton儲(chǔ)存?式:4°C,sealedstorage,awayfrommoisture*Insolvent:-80°C,6months;-20°C,1month(sealed

storage,awayfrommoisture)溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:25mg/mL(50.71mM;ultrasonicandwarmingandheatto80°C)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.0286mL10.1428mL20.2856mL5mM0.4057mL2.0286mL4.0571mL10mM0.2029mL1.0143mL2.0286mL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(sealedstorage,awayfrommoisture)。-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:≥2.5mg/mL(5.07mM);Clearsolution2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(5.07mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(5.07mM);ClearsolutionBIOLOGICALACTIVITY?物活性G-5555有效PAK1抑制劑，對(duì)PAK1和PAK2的Ki值分別為3.7nM和11nM。IC50&TargetPAK1PAK23.7nM(Ki)11nM(Ki)體外研究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].體內(nèi)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?邊的抑制劑?師www.MedChemEstoredat-70to-80°Cuntilanalysis[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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.