GW-6471-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemEGW6471Cat.No.:HY-15372CASNo.:880635-03-0分?式:C??H??F?N?O?分?量:619.67作?靶點(diǎn):PPAR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:83.33mg/mL(134.47mM;Needultrasonic)掃描?維碼，H2O:<0.1mg/mL(insoluble)運(yùn)?溶解?案計(jì)算器獲得適合您實(shí)驗(yàn)體系的溶解?案MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM1.6138mL8.0688mL16.1376mL5mM0.3228mL1.6138mL3.2275mL10mM0.1614mL0.8069mL1.6138mL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，并請(qǐng)注意儲(chǔ)備液的保存?式和期限。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?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶1.請(qǐng)依序添加每種溶劑：10%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.08mg/mL(3.36mM);Clearsolution此?案可獲得≥2.08mg/mL(3.36mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲(chǔ)備液加到400μLPEG300中，混合均勻；向上述體系中加?50μLTween-80，混合均勻；然后繼續(xù)加?450μL?理鹽?定容?1mL。1/4www.MedChemEwww.MedChemE2.請(qǐng)依序添加每種溶劑：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:2.08mg/mL(3.36mM);Suspendedsolution;Needultrasonic此?案可獲得2.08mg/mL(3.36mM)的均勻懸濁液，懸濁液可?于?服和腹腔注射。以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲(chǔ)備液加到900μL20%的SBE-β-CD?理鹽??溶3.液中，混合均勻。請(qǐng)依序添加每種溶劑：10%DMSO90%cornoilSolubility:≥2.08mg/mL(3.36mM);Clearsolution此?案可獲得≥2.08mg/mL(3.36mM，飽和度未知)的澄溶液，此?案不適?于實(shí)驗(yàn)周期在半個(gè)?以上的實(shí)驗(yàn)。以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲(chǔ)備液加到900μL??油中，混合均勻。BIOLOGICALACTIVITY?物活性GW6471?種有效的PPARα拮抗劑。IC50&TargetPPARα體外研究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].體內(nèi)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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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