GSK461364-GSK461364A-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEGSK461364Cat.No.:HY-50877CASNo.:929095-18-1Synonyms:GSK461364A分?式:C??H??F?N?O?S分?量:543.6作?靶點(diǎn):Polo-likeKinase(PLK)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:50mg/mL(91.98mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM1.8396mL9.1979mL18.3959mL5mM0.3679mL1.8396mL3.6792mL10mM0.1840mL0.9198mL1.8396mL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%saline1/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:≥2.5mg/mL(4.60mM);Clearsolution2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(4.60mM);Clearsolution3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.60mM);ClearsolutionBIOLOGICALACTIVITY?物活性GSK461364選擇性，可逆，ATP競(jìng)爭(zhēng)性的PLK1抑制劑，Ki值為2.2nM。IC50&TargetPLK12.2nM(Ki)體外研究GSK461364isapotent,selective,andreversibleATP-competitivePlk1inhibitor(Ki,2.2nM)withatleast390-foldgreaterselectivityforPlk1thanforPlk2andPlk3and1,000-foldgreaterselectivitythanforapanelof48otherkinases[1].GSK461364(GSK461364A,250nM)inhibitingplk1causesprolongedmitoticdelay,aberrantmitoticexit,andp53activationinA549andPL45cells.Knockdownofp53significantlyenhancesthesensitivityofthecellstoGSK461364A(30or300nM)inpreventingoutgrowthinA549andNCI-H460cells,comparedwithcellswithnonsilencingcontrolsiRNA[2].GSK461364caninhibitscellgrowthofmostproliferatingcancercelllines,andsuppresses89%ofcancercellline(66of74lines)proliferation,withaGI50(concentrationrequiredtoinhibit50%cellgrowth)of≤100nM.GSK461364(GSK461364A,>20nM)blockscellsinG2-MphasewithreductionofcellsinG1phaseofA549lungcarcinomaline.GSK461364(10-250nM)blockscellsinG2andMphasesofthecellcycleandcausesM-phasecaspase-3/caspase-7activationincancercells[3].GSK461364(0.5-2μM)decreaseslevelofPLK1andpCDC25C,andcuasesadose-andtime-dependentincreaseinpHisH3,anindicatorofmitoticarrestinOScelllines[4].體內(nèi)研究GSK461364(50mg/kg)exhibitsvariousdegreesoftumorgrowthdelay(TGD)inmultiplexenografttumormodelsbyi.p.onedoseevery2daysrepeatedtwelvetimes(q2d×12)[1].PROTOCOLKinaseAssay[3]Kinasereactionsareperformedinafinalassayvolumeof10μLusingtheZ'-LyteAssaykit(Ser/Thrpeptide16).Briefly,reactionscontained50mMHEPES(pH7.5),10mMMgCl2,1mMEGTA,1mMDTT,0.01%Brij35,0.01mg/mLcasein,200μMATP,200μMPoloBoxpeptide(NH2-MAGPMQS[pT]PLNGAKK-OH),and6nMrecombinantPlk1(H6-tev-PLK1-603).Plk1ispreincubatedfor60mininthepresenceorabsenceof0to1,000nMGSK461364.Reactionsaretheninitiatedbytheadditionof2μMpeptide.After15minat23°C,reactionsarequenchedandprocessedaccordingtheZ'-Lyteprotocolandreadonanplatereader.Rawfluorescencevaluesareconvertedtoconcentrationofproductformedusingsubstrateandproductstandards.IC50valuesaredeterminedusingatwo-parameterfit(HillcoefficientandIC50)usingGraFitsoftware.BecausethepotencyofinhibitionforGSK461364isobservedtovaryasafunctionoftheATPconcentrationinamannerconsistentwithanATP-competitivemodeofinhibition,anupperlimitfortheKi*appforGSK461364isdeterminedbyapplyingtheCheng-Prusoffrelationshipforacompetitiveinhibitor(ATP2/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEKm*app=16μM)totheIC50valueobtainedwith60minpreincubationofGSK461364.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[3]Celllinesgrowintherecommendedmediaat37°C,5%CO2inahumidifiedincubator.Cellsareplatedintriplicate96-wellmicrotiterplatesat1,000cellsperwellinculturemedia.GSK461364(GSK461364A)dissolvedinDMSOornegativecontrol(0.1%DMSO)areaddedthefollowingday,andoneplateisharvestedwith50μLofCellTiter-Gloforatime0(T=0)measurement.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalCellsareimplantedinNudemiceandgrownastumorxenografts.DosingbeganwhentumorsachieveapprAdministration[3]100mm3.GSK461364(GSK461364A)orthevehicle[4%DMA/Cremaphore(50:50),pH5.6]isgiveni.p.tomiceevery2d(q2d×6,q2d×12)orevery4d(q4d×3)atnominaldoselevelsof25,50,and100mg/kg/dose.Resultsarereportedasmediantumorvolumeforn=7to8mice.Paclitaxel(30mg/kgi.v.;q4d×3)isusedasapositivecontrolforcomparison.TumorsaremeasuredthriceaweekwithVerniercalipers,andtumorvolumeiscalculatedfromtwo-dimensionalmeasurementsusinganequationapproximatingthevolumeofaprolateellipsoid[tumorvolumemm3=(length×width2)×0.5].Themaximumtolerateddoseisdefinedasthehighestdosethatproduces>20%mortalityor>20%weightloss(appr4g).Antitumoractivityisdefinedastumorgrowthdelay(TGD),partialregression(PR),orcompleteregression(CR).TGDrepresentsthetimedifferentialbetweenthetreatedandcontroltumorstoreachapredeterminedtumorvolumeof1,000mm3.PRisdefinedasadecreaseinanindividualtumorvolumetoone-halftheinitialstartingvolumeforatleast1wk(threeconsecutivemeasurements).CRisdefinedasadecreaseinanindividualtumorvolumeto3foratleast1wk.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?MolCell.2022May25;S1097-2765(22)00443-9.?SciTranslMed.2018Jul18;10(450).pii:eaaq1093.?Theranostics.2022;12(8):3911-3927.?EurJCancer.2013Sep;49(14):3020-8.?JCellSci.2019Jul1;132(13):jcs229385.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].OlmosD,etal.PhaseIstudyofGSK461364,aspecificandcompetitivePolo-likekinase1inhibitor,inpatientswithadvancedsolidmalignancies.ClinCancerRes.2011May15;17(10):3420-30.[2].DegenhardtY,etal.SensitivityofcancercellstoPlk1inhibitorGSK461364Aisassociatedwithlossofp53functionandchromosomeinstability.M