Torkinib-PP-242-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemETorkinibCat.No.:HY-10474CASNo.:1092351-67-1Synonyms:PP242分⼦式:C₁₆H₁₆N₆O分⼦量:308.34作⽤靶点:mTOR;Autophagy;Mitophagy;Apoptosis作⽤通路:PI3K/Akt/mTOR;Autophagy;Apoptosis储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:50mg/mL(162.16mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM3.2432mL16.2159mL32.4317mL5mM0.6486mL3.2432mL6.4863mL10mM0.3243mL1.6216mL3.2432mL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%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:≥2.5mg/mL(8.11mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(8.11mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(8.11mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Torkinib(PP242)⼀种选择性，ATP竞争型的mTOR抑制剂，IC50为8nM。PP242抑制mTORC1和mTORC2的IC50分别为30nM和58nM。IC50&TargetmTORC1mTORC2mTORp110δ30nM(IC50)58nM(IC50)8nM(IC50)100nM(IC50)DNA-PKPDGFRp110αp110β410nM(IC50)410nM(IC50)2μM(IC50)2.2μM(IC50)p110γAblHckScr1.3μM(IC50)3.6μM(IC50)1.2μM(IC50)1.4μM(IC50)Scr(T338I)VEGFR2EGFREphB45.1μM(IC50)1.5μM(IC50)4.4μM(IC50)3.4μM(IC50)AutophagyMitophagy体外研究Torkinib(PP242)potentlyinhibitsmTOR(IC50=8nM)butismuchlessactiveagainstotherPI3Kfamilymembers.TestingofTorkinib(PP242)against219proteinkinasesrevealsremarkableselectivityrelativetotheproteinkinome:ataconcentration100-foldaboveitsIC50formTOR,Torkinib(PP242)inhibitsonlyonekinasebymorethan90%(Ret)andonlythreebymorethan75%(PKCα,PKCβIIandJAK2V617F)[1].Torkinib(PP242)hasadose-dependenteffectonproliferationandathigherdosesismuchmoreeffectivethanRapamycinatblockingcellproliferation.TheabilityofTorkinib(PP242)toblockcellproliferationmoreefficientlythanRapamycincouldbearesultofitsabilitytoinhibitmTORC1andmTORC2,becauseRapamycincanonlyinhibitmTORC1.InSIN1-/-mouseembryonicfibroblasts(MEFs),RapamycinisalsolesseffectiveatblockingcellproliferationthanTorkinib.ThatTorkinib(PP242)andRapamycinexhibitverydifferentanti-proliferativeeffectsinSIN1-/-MEFssuggeststhatthetwocompoundsdifferentiallyaffectmTORC1[2].体内研究Infatandliver,Torkinib(PP242)isabletocompletelyinhibitthephosphorylationofAktatS473andT308,consistentwithitseffectonthesephosphorylationsitesobservedincellculture.Surprisingly,Torkinib(PP242)isonlypartiallyabletoinhibitthephosphorylationofAktinskeletalmuscleandismoreeffectiveatinhibitingthephosphorylationofT308thanS473,despiteit'sabilitytofullyinhibitthephosphorylationof4EBP1andS6.Theseresultswillbeconfirmedbyinvivodose-responseexperiments,but,consistentwiththepartialeffectofTorkinib(PP242)onpAktinskeletalmuscle,amuscle-specificknockoutoftheintegral2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEmTORC2componentrictorresultedinonlyapartiallossofAktphosphorylationatS473.TheseresultssuggestthatakinaseotherthanmTOR,suchasDNA-PK,maycontributetophosphorylationofAktinmuscle[2].PROTOCOLCellAssay[2]Wild-typeandSIN1-/-MEFsareplatedin96-wellplatesatapproximately30%confluenceandleftovernighttoadhere.ThefollowingdaycellsaretreatedwithTorkinib(PP242)(1nM,10nM,100nM,1μM,and10μM),Rapamycin,orvehicle(0.1%DMSO).After72hoftreatment,10μLof440μMresazurinsodiumsaltisaddedtoeachwell,andafter18h,theflorescenceintensityineachwellismeasuredusingatop-readingflorescentplatereaderwithexcitationat530nmandemissionat590nm[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[2]Administration[2]Six-wk-oldmaleC57BL/6micearefastedovernightpriortodrugtreatment.Torkinib(PP242)(0.4mg),Rapamycin(0.1mg),orvehiclealoneisinjectedIP.After30minfortheRapamycin-treatedmouseor10minfortheTorkinib(PP242)andvehicle-treatedmice,250mUofinsulinin100μLofsalineisinjectedIP.15minaftertheinsulininjection,themicearekilledbyCO2asphyxiationfollowedbycervicaldislocation.Tissuesareharvestedandfrozenonliquidnitrogenin200μLofcaplysisbuffer.Thefrozentissueisthawedonice,manuallydisruptedwithamortarandpestle,andthenfurtherprocessedwithamicrotissue-homogenizer.ProteinconcentrationoftheclearedlysateismeasuredbyBradfordassayand5-10μgofproteinisanalyzedbyWesternblot[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•ActaPharmSinB.2020Jun;10(6):1004-1019.•CancerRes.2013Apr15;73(8):2574-86.•JExpClinCancerRes.2021Jan9;40(1):25.•Theranostics.2022Jan1;12(2):675-688.•BreastCancerRes.2020Jun3;22(1):59.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].ApselB,etal.Targetedpolypharmacology:discoveryofdualinhibitorsoftyrosineandphosphoinositidekinases.NatChemBiol.2008Nov;4(11):691-9.[2].FeldmanME,etal.Active-siteinhibitorsofmTORtargetrapa