PDD-00017273-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEPDD00017273Cat.No.:HY-108360CASNo.:1945950-21-9分⼦式:C₂₃H₂₆N₆O₄S₂分⼦量:514.62作⽤靶点:Poly(ADP-ribose)Glycohydrolase(PARG)作⽤通路:CellCycle/DNADamage储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:25mg/mL(48.58mM;ultrasonicandwarmingandheatto60°C)MassSolvent1mg5mg10mgConcentration制备储备液1mM1.9432mL9.7159mL19.4318mL5mM0.3886mL1.9432mL3.8864mL10mM0.1943mL0.9716mL1.9432mL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%salineSolubility:≥2.5mg/mL(4.86mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(4.86mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.86mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性PDD00017273⼀种有效的聚(腺苷⼆磷酸核糖)⽔解酶(PARG)抑制剂，IC50值为26nM，KD值为1.45nM。IC50&TargetIC50:26nM(PARG)[1]KD:1.45nM(PARG)[1]体外研究PDD00017273isapotentinhibitorofPARG,withanIC50of26nM,andaKDof1.45nM.PDD00017273(10μM)doesnotinhibitfivecommonCytochromeP450enzymes.PDD00017273(30μM)modestlyincreasessphosphorylatedH2AX(γH2AX)intensity,PDD00017273alsodecreasesinNAD/HthroughPARGinhibitionafterDNAdamage.PDD00017273suppressestheZR-75-1cellscarringBRCA1andBRCA2wildtype,andexhibitslesspotentactivitiesagainstMDA-MB-436cellscarrythe5396+1G>AmutationinBRCA1[1].PDD00017273(0.3μM)inhibitsdegradationofPARpolymersinMCF7cells.PDD00017273(0.3μM)alsoreducestheviabilityofBRCA1,BRCA2,PALB2,FAM175A,andBARD1depletedcells.PDD00017273stallsreplicationforksandinducesDNAdamagethatrequireshomologousrecombination(HR)forrepair[2].PROTOCOLKinaseAssay[1]Briefly,PARGinvitroassaysareconductedinatotalvolumeof15μLinastandard384-wellformat.Atotalof5μLofhumanfulllengthPARGusedatafinalreactionconcentrationof65pM,isaddedto5μLofBt-NADribosylatedPARP1substrateatafinalreactionconcentrationof4.8nMinassaybuffer(50mMTrispH7.4,0.1mg/mLBSA,3mMEDTA,0.4mMEGTA,1mMDTT,0.01%Tween20,50mMKCl).ThereactionisincubatedatRTfor10min,andthen5μLofdetectionreagentisadded.Detectionreagentconsistsof42nMmAbanti-6HISXL665and2.25nMstreptavidineuropiumcryptate,bothat3×workingstockconcentrations(finalconcentrationsof14nMand0.75nM,respectively),indetectionbuffer(50mMTrispH7.4,0.1mg/mLBSAand100mMKF).FollowingincubationatRTfor60mininthedark,TR-FRETsignalismeasuredatλEx340nmandλEm665nmandλEm620nmusingaPHERAstarFSplatereader.Theratioiscalculatedas[Em665/EM620]×104foreachwellandusedtocalculatepercentinhibitionfortestcompounds[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]HeLacellsareseededin30μLofmediaat1×104cells/mLinGreiner384-wellplates.Atotalof16-24hlater,cellsaretreatedwithinhibitors(8ptdoseresponse,0.01-30μM,triplicates)orvehicle(DMSO)control.Theouterwellsareleftundosedtoaccountforedgeeffects.After72h,50μLof3.7%formaldehyde/PBSisaddedtoeachwell,andcellsarefixedfor20min.CellsarethenrinsedtwicewithPBSandstainedfor1hwithHoechst33342/PBS(1:2000)inthedark.AftertwofurtherrinseswithPBS,imagesarecapturedand2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEnucleicountedonaCellInsight.Themaximumnumberoffieldsarecapturedfromeachtriplicatewell,whichapproximatedtoatleast1000nucleiinvehicle-dosedwells[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CellRep.2021Oct5;37(1):109695.•Elife.2022Apr27;11:e72464.•Viruses.2022Sep15;14(9):2049.•CancerRep.26August2022.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].JamesDI,etal.First-in-ClassChemicalProbesagainstPoly(ADP-ribose)Glycohydrolase(PARG)InhibitDNARepairwithDifferentialPharmacologytoAZD2281.ACSChemBiol.2016Nov18;11(11):3179-3190.Epub2016Oct12.[2].GravellsP,etal.SpecifickillingofDNAdamage-responsedeficientcellswithinhibitorsofpoly(ADP-ribose)glycohydrolase.DNARe