GW-6471-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemEGW6471Cat.No.:HY-15372CASNo.:880635-03-0分⼦式:C₃₅H₃₆F₃N₃O₄分⼦量:619.67作⽤靶点:PPAR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:83.33mg/mL(134.47mM;Needultrasonic)扫描⼆维码，H2O:<0.1mg/mL(insoluble)运⽤溶解⽅案计算器获得适合您实验体系的溶解⽅案MassSolvent1mg5mg10mgConcentration制备储备液1mM1.6138mL8.0688mL16.1376mL5mM0.3228mL1.6138mL3.2275mL10mM0.1614mL0.8069mL1.6138mL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，并请注意储备液的保存⽅式和期限。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%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.08mg/mL(3.36mM);Clearsolution此⽅案可获得≥2.08mg/mL(3.36mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到400μLPEG300中，混合均匀；向上述体系中加⼊50μLTween-80，混合均匀；然后继续加⼊450μL⽣理盐⽔定容⾄1mL。1/4www.MedChemEwww.MedChemE2.请依序添加每种溶剂：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:2.08mg/mL(3.36mM);Suspendedsolution;Needultrasonic此⽅案可获得2.08mg/mL(3.36mM)的均匀悬浊液，悬浊液可⽤于⼝服和腹腔注射。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL20%的SBE-β-CD⽣理盐⽔⽔溶3.液中，混合均匀。请依序添加每种溶剂：10%DMSO90%cornoilSolubility:≥2.08mg/mL(3.36mM);Clearsolution此⽅案可获得≥2.08mg/mL(3.36mM，饱和度未知)的澄溶液，此⽅案不适⽤于实验周期在半个⽉以上的实验。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL⽟⽶油中，混合均匀。BIOLOGICALACTIVITY⽣物活性GW6471⼀种有效的PPARα拮抗剂。IC50&TargetPPARα体外研究Inacell-basedreporterassay,GW6471completelyinhibitsGW409544-inducedactivationofPPARαwithanIC50of0.24μM[1].ThefunctionalroleofPPARαisevaluatedonrenalcellcarcinoma(RCC)cellviabilitybyMTTassay.BothCaki-1(VHLwildtype)and786-O(VHLmutated)cellsareincubatedseparatelywithaspecificPPARαagonist,WY14,643,oraspecificPPARαantagonist,GW6471atconcentrationsfrom12.5to100µMfor72hours,andcellviabilityisassessed.WhileWY14,643eitherhasnoaffecton,orslightlyincreased,cellviability,GW6471significantlyanddose-dependentlyinhibitscellviability(uptoapproximately80%)inbothcelllines[2].体内研究TotesttheantitumoractivityofPPARαantagonisminvivo,asubcutaneousxenograftmousemodelisused.Caki-1cellsareimplantedsubcutaneouslyinnude(Nu/Nu)mice.Aftertumormassesreach-5mmindiameter,GW6471isadministratedintraperitoneallyeveryotherdayfor4wkatadose(20mg/kgmousebodywt)thatisdescribedtobeeffectiveinaninvivodose-responsestudyandconfirmedheretobeefficacious.Therearesignificantdifferencesintumorgrowthbetweenvehicle-andGW6471-treatedanimals.NotoxicityisobservedatthedosesofGW6471basedonweightsoftheanimals,andlaboratoryvalues,includingkidneyandliverfunctiontests,arenotadverselyaffected.Todemonstrateon-targeteffectsofGW6471,c-Myclevelsareevaluatedinthetumors,whichshowsignificantdecreasesintheGW6471-treatedanimals[3].PROTOCOLCellAssay[2]786-OandCaki-1cellsareplatedin96wellplates.BothcellsareincubatedseparatelywithWY14,643orGW6471atconcentrationsfrom12.5to100µMfor72hours,andaftertheindicatedtreatments,thecellsareincubatedinMTTsolution/mediamixture.Then,theMTTsolutionisremovedandthebluecrystallineprecipitateineachwellisdissolvedinDMSO.Visibleabsorbanceofeachwellat540nmisquantifiedusingamicroplatereader[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.2/4www.MedChemEwww.MedChemEAnimalMice[3]Administration[3]MaleathymicNu/Numice(8wkofage,~25gbodywt)areinjectedwith1×105Caki-1cellssubcutaneously(3:1DMEM-Matrigel)intheflankregion.Tumorprogressionismonitoredweeklybycalipers.Whentumorsizereaches~80-100mm3,animalsarerandomlyassignedtofourgroupsandtreatmentsarestarted(day1).ThevehiclegroupreceiveDMSO(4%inPBS)intraperitoneallyandvegetableoilviaoralgavage.ThePPARαgroupisinjectedintraperitoneallywithGW6471inthesamevehicle(20mg/kgbodywt;murinedoseresponseisreportedelsewhere)everyotherday.TheSunitinibgroupreceiveSunitinibinvegetableoilviaoralgavage(40mg/kgbodywt)5days/wk.AnothergroupreceiveGW6471+Sunitinib.Todetermineanypotentialtoxicityofthetreatment(s),bodyweightsoftheanimalsaremeasuredandsignsofadversereactionsaremonitored.Onday28,themiceareeuthanizedandthetumormassisdetermined.Tumorgrowthrateiscalculated[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•Gut.2020Nov30;gutjnl-2020-321774.•JCellPhysiol.2021Mar;236(3):1889-1902.•OxidMedCellLongev.2021Feb26.•OxidMedCellLongev.2019Nov3;2019:7536803.•JCellMolMed.2020Mar;24(6):3384-3398.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].XuHE,etal.Structuralbasisforantagonist-mediatedrecruitmentofnuclearco-repressorsbyPPARalpha.Nature.2002Feb14;415(6873):813-7.[2].AbuAboudO,etal.InhibitionofPPARαinducescellcyclearrestandapoptosis,andsynergizeswithglycolysisinhibitioninkidneycancercells.PLoSOne.2013Aug7;8(8):e71115.[3].AbuAboudO,etal.PPARαinhibitionmodulatesmultiplereprogrammedmetabolicpathwaysinkidneycancerandattenuatestumorgrowth.AmJPhysiolCellPhysiol.2015Jun1;308(11):C890-8.McePdfHeight3/4www.M