DHW-221-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors • ScreeningLibraries • Proteinswww.MedChemEDHW-221Cat.No.:HY-130133CASNo.:2378831-21-9分子式:C₂₇H₂₂F₂N₄O₅S分子量:552.55作用靶点:PI3K;mTOR;Akt;Apoptosis;Paraptosis;p38MAPK;MitochondrialMetabolism;P-glycoprotein;CDK;MMP;HIF/HIFProlyl-Hydroxylase;VEGFR作用通路:PI3K/Akt/mTOR;Apoptosis;MAPK/ERKPathway;MetabolicEnzyme/Protease;MembraneTransporter/IonChannel;CellCycle/DNADamage;ProteinTyrosineKinase/RTK储存方式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY生物活性DHW-221是一种口服有效的PI3K/mTOR双重抑制剂，对四种I类PI3K亚型以及mTOR(PI3Kα，IC50=0.50nM；PI3Kβ，IC50=1.9nM；PI3Kγ，IC50=1.8nM；PI3Kδ，IC50=0.74nM;mTOR,IC50=3.9nM)均表现出低纳米摩尔级别的效力。DHW-221通过阻断PI3K/Akt/mTOR通路、诱导线粒体(mitochondrial)凋亡(apoptosis)和副凋亡(paraptosis)(通过内质网应激和MAPK信号传导)以及引起细胞周期阻滞，进而抑制细胞迁移、侵袭和血管生成，从而发挥抗肿瘤作用。DHW-221在A549/Taxol(HY-B0015)和HCC827小鼠模型中均抑制肿瘤生长。DHW-221可用于非小细胞肺癌(NSCLC)、结肠癌和乳腺癌的研究[1][2][3]。IC50&TargetPI3KαPI3KβPI3KδPI3Kγ0.50nM(IC50)1.9nM(IC50)0.74nM(IC50)1.8nM(IC50)mTOR3.9nM(IC50)体外研究DHW-221(compound8i)(72h)showsantiproliferationactivityagainsttypicallyhumancancercellswithIC50valuesof0.36μM(T47D),0.14μM(HCT116),and0.31μM(MCF-7)[1].DHW-221(0.3-3μM)decreasesthephospho-Akt(S473)inadose-dependentmannerinHCT116cellswithnosignificantchangeintheexpressionoftotalproteinAkt[1].DHW-221(0-5μM,2weeks)inhibitstheproliferationofHCT116cellsinadose-dependentmanner[1].DHW-221(0.3-3μM,0-48h)inhibitsHCT116cellmetastasisandinvasivenessinadose-dependentmanner[1].1/7 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEDHW-221(0.3-3μM)inducesapoptosisinHCT116cellsinaconcentration-dependentmanner,withmarkednuclearfragmentationandcondensationofchromatin[1].DHW-221fitsintothebindingsiteofPI3KkinasesimilarlytoOmipalisib(HY-10297),forminghydrogenbondswithVal882,Lys833,andThr887,aswellasPi-Piinteractionswithmultipleaminoacidresidues[1].DHW-221(72h)exhibitssignificantcytotoxicityinaconcentration-andtime-dependentmannerinA549/Taxol(IC50=0.5274μM)andA549cells(IC50=0.4242μM)[2].DHW-221(0-2.4μM,48-72h)exertssignificantinhibitoryactivityinMDRcancercells(A549andA549/Taxolcells),resultinginseverecellulardamageat48hcomparedtocontrolgroupinbothcells[2].DHW-221(0.15-2.4μM,48h)increasesintracellularRho-123accumulationinaconcentration-dependentmannerinA549/TaxolcellsandsignificantlydownregulatesP-gpexpression,indicatingthatitinhibitsP-gpfunctionandproteinexpression[2].DHW-221(50nM,48h)significantlyenhancesthecytotoxiceffectofTaxol,aneffectsimilartotheP-gpinhibitorVerapamil(HY-14275),suggestingthatitfunctionsasaP-gpinhibitorandaMDRreversalagent[2].DHW-221(0.15-2.4μM,12-48h)triggersapoptosisandparaptosisinA549/Taxolcellsthroughthemitochondrialpathway,ERstressandtheMAPKsignalingpathway[2].DHW-221(0.15-2.4μM,48h)arreststhecellcycleattheG0/G1phaseinA549/TaxolandA549cellsbydownregulatingcyclinD1,CDK4,andCDK6,andupregulatingp21[2].DHW-221(0.05-0.15μM,48h)suppressesthemigrationandinvasioncapabilitiesofA549/Taxolcellsthroughreversingepithelial-mesenchymaltransition(EMT)phenotypicchanges[2].DHW-221(0.15-2.4μM,48h)significantlyblocksthePI3K/AktsignalingpathwayinbothA549andA549/Taxolcells,asdemonstratedbydecreasedlevelsofPI3Kp110αandphosphorylatedAkt(p-Akt)[2].DHW-221(25-400nM,1-3h)inhibitsendothelialtubeformationinHumanUmbilicalVeinEndothelialCells(HUVECs)bysuppressingthePI3K/HIF-1α/VEGFsignalingaxis[3].DHW-221(1.56-6.25µM)inhibitsmicrovesselsproutinginrataorticringassaysexvivo[3].DHW-221(1.25-5µM)inhibitsneovascularizationinchickchorioallantoicmembraneswithoutaffectingembryoviability[3].CellProliferationAssay[1]CellLine:HCT116cellsConcentration:0.3,1and3μMIncubationTime:2weeksResult:InhibitedtheformationofHCT116cellclonesinaconcentration-dependentmanner.Almostcompletelyinhibitedtheproductionofcellclonesat3μM.CellMigrationAssay[1]CellLine:HCT116cellsConcentration:0.3,1and3μMIncubationTime:0,24and48h2/7 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEResult:Itsinhibitionabilityincreasedinaconcentration-dependentmanner.CellInvasionAssay[1]CellLine:HCT116cellsConcentration:0.3,1and3μMIncubationTime:48hResult:SignificantlyinhibitedHCT116cellmetastasisandinvasivenessinadose-dependentmanner.CellViabilityAssay[2]CellLine:A549andA549/TaxolcellsConcentration:0.075,0.15,0.3,0.5,1.2and2.4μMIncubationTime:24,48and72hResult:Exhibitedsignificantcytotoxicityinaconcentration-andtime-dependentmannerinA549/TaxolandA549cellswithanresistanceindex(RI)of1.2.CellProliferationAssay[2]CellLine:A549andA549/TaxolcellsConcentration:0.15,0.60and2.40μMIncubationTime:2weeksResult:DecreasedthecolonyformationabilityofA549/TaxolandA549cellsinaconcentration-dependentmanner.CellCytotoxicityAssay[2]CellLine:A549andA549/TaxolcellsConcentration:0.15,0.60and2.40μMIncubationTime:48hResult:IncreasedLDHreleaseratesinA549andA549/Taxolcellsinaconcentration-dependentmanner.WesternBlotAnalysis[2]CellLine:A549andA549/Taxolcells3/7 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEConcentration:0.15,0.60and2.40μMIncubationTime:3,6,12,24and48hResult:Upregulatedtheexpressionlevelsofapoptosis-relatedproteins(cytochromeC,cleavedcaspase3,andcleavedPARP)inA549/Taxolcellsinaconcentration-dependentmanner.Downregulatedtheexpressionlevelsoftheanti-apoptoticprotein,Bcl-2,inA549/Taxolcellsinaconcentration-dependentmanner.SignificantlyinhibitedtheexpressionofAlixinA549/Taxolcells.SignificantlyupregulatedtheexpressionlevelsofATF4,CHOP(keyregulatorinERstress),p-JNK,p-ERK,andp-p38inA549/Taxolcells.DecreasedtheexpressionlevelsofcyclinD1,CDK4,andCDK6andincreasedp21levelsinA549/Taxolcellsinaconcentration-dependentmanner.SignificantlydecreasedtheexpressionlevelsofPI3Kp110aandp-Aktinaconcentration-dependentmanner,withnoobviouschangeinthetotalAktlevelinbothcells.BlockedthePI3K/AktsignalingpathwayinbothA549/TaxolandA549cells.Significantlydownregulatedp-FOXO3a(Ser253)expressionandupregulatedFOXO3aexpression,accompaniedbyanincreaseinBimexpressioninA549/Taxolcells.ShowednosignificantchangesinFOXO3aandBimexpressionlevelswereobservedinA549cells.PromotedFOXO3aaccumulationinA549/TaxolcellswhencombinedwithMG132(HY-13259).InterferedwithFOXO3adegradationinaproteasome-independentmanner.Immunofluorescence[2]CellLine:A549/TaxolcellsConcentration:0.15,0.60and2.40μMIncubationTime:24and48hResult:IncreasednuclearFOXO3aproteinanddecreasedcytoplasmicphosphorylatedFOXO3aattheSer253site.ReversedtheEMTphenotype,evidencedbyenhancedfluorescenceintensityoftheepithelialmarkeroccludinandweakenedfluorescenceofthemesenchymalmarkervimentin.ApoptosisAnalysis[2]CellLine:A549andA549/TaxolcellsConcentration:0.15,0.60and2.40μM4/7 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEIncubationTime:12h(forvacuolation),and48h(forapoptosis/MMP)Result:SignificantlyincreasedtheratesofearlyandlateapoptosisinbothA549/TaxolandA549cellsafter48h.Itspro-apoptoticabilityinA549/TaxolcellwasstrongerthanthatofA549cellsat2.4μM.TriggeredapoptosisinA549/TaxolandA549cellsinaconcentration-dependentmanner.DecreasedtheintracellularMMPinA549/Taxolcellsafter48h.InducedtheformationofmassivevacuolesinA549/Taxolcellsat2.4μMafter12h.AttenuatedcytoplasmicvacuolationwhencombinedwithCycloheximide(HY-12320,20μM).CellCycleAnalysis[2]CellLine:A549/TaxolcellsConcentration:0.15,0.60and2.40μMIncubationTime:48hResult:SignificantlyreducedtheproportionofcellsinSphaseandincreasedtheproportionofcellsinG0/G1phase.CellMigrationAssay[2]CellLine:A549andA549/TaxolcellsConcentration:0.05,0.10and0.15μMIncubationTime:0,24and48hResult:SignificantlydecreasedthewoundhealingratecomparedtothecontrolgroupinbothA549andA549/Taxolcells.CellInvasionAssay[2]CellLine:A549andA549/TaxolcellsConcentration:0.05,0.10and0.15μMIncubationTime:24hResult:SignificantlyreducedthenumberofcellsthatmigratedandinvadedthroughthechamberinA549/Taxolcells.WesternBlotAnalysis[2]CellLine:A549/Taxolcells5/7 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEConcentration:0.05,0.10and0.15μMIncubationTime:48hResult:IncreasedtheexpressionlevelsofE-cadherinandoccludinbutsignificantlydecreasedtheexpressionlevelsofmesenchymalmarkers,includingvimentinandslug.Decreasedtheexpressionoftranscriptionfactorsnail.SignificantlydownregulatedtheexpressionlevelsofMMP2andMMP9inA549/Taxolcells.体内研究DHW-221(10,20,and40mg/kg,i.g.,q.d.for2weeks)inhibitstumorgrowthwithoutbodyweightchangeandtoxicitythroughFOXO3atranslocationinA549/Taxolcell-bearingmice[2].DHW-221(10,20,and40mg/kg,p.o.,q.d.for21days)inhibitstumorgrowthinHCC827xenograftmicemodel[3].AnimalModel:Malenudemice(4-6weeksold)intravenouslyinjectedwithA549/Taxolcells[2]Dosage:25and50mg/kgAdministration:i.g.,q.d.for2weeksResult:Significantlydecreasedthenumberoflungnodules.Downregulatedp-FOXO3aexpressionandupregulatedFOXO3aexpression.ShowednochangeinKi67expression.Showednosignificantdifferencesinthebodyweightandvisceraindexcomparedtothecontrolgroup.Causednobehavioralabnormalitiesorlossofappetite.Exhibitednoevidenthistologicalabnormalitiesintheheart,liver,kidney,orspleen.Resultedinnoabnormaltoxicitysignsinserumbiochemicalparameters(CK,ALT,AST,CRE),indicatingnocardiac,hepatic,orrenaltoxicity.AnimalModel:NudemicebearingHCC827xenografts(Subcutaneousandorthotopicinjection)[3]Dos