Perifosine-KRX-0401-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEPerifosineCat.No.:HY-50909CASNo.:157716-52-4Synonyms:KRX-0401;NSC639966;D21266分?式:C??H??NO?P分?量:461.66作?靶點:Akt;Autophagy;Apoptosis作?通路:PI3K/Akt/mTOR;Autophagy;Apoptosis儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗H2O:≥153.33mg/mL(332.13mM)DMF:<1mg/mL(insoluble)DMSO:<1mg/mL(insolubleorslightlysoluble)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.1661mL10.8305mL21.6610mL5mM0.4332mL2.1661mL4.3322mL10mM0.2166mL1.0830mL2.1661mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE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.請依序添加每種溶劑：PBSSolubility:50mg/mL(108.30mM);Clearsolution;NeedultrasonicBIOLOGICALACTIVITY?物活性Perifosine?種有?服活性的Akt抑制劑，抑制不同腫瘤細(xì)胞系增殖的IC50值為0.6-8.9μM。IC50&TargetAutophagy體外研究TheIC50forgrowthofNtv-a/LacZcelllinesisdeterminedbyMTTassay.Whenthecellsareculturedfor48hoursin10%FCS-supplementedmedia,theIC50forcellswithconstitutivelyactivePDGF,Ras,orAktsignalingissimilarandfoundtobe~45μM[1].Perifosine,aoral-bioavailablealkylphospholipid(ALK),onthecellcyclekineticsofimmortalizedkeratinocytes(HaCaT)aswellasheadandnecksquamouscarcinomacells.Proliferationisassessedbytheincorporationof[3H]thymidineintocellularDNA.ExposuretoPerifosine(0.1-30μM)for24hresultsinadose-dependentinhibitionof[3H]thymidineuptakeinallcelllinestested.TheIC50sforgrowtharebetween0.6and8.9μM,reachingIC80sof~10μM.PerifosineblockscellcycleprogressionofheadandnecksquamouscarcinomacellsatG1-SandG2-Mbyinducingp21WAF1,irrespectiveofp53function,andmaybeexploitedclinicallybecausethemajorityofhumanmalignanciesharborp53mutations.Perifosine(20μM)inducesbothG1-SandG2-Mcellcyclearrest,togetherwithp21WAF1expressioninbothp53wild-typeandp53-/-clones[2].體內(nèi)研究Miceareidentifiedwithtumorsbybioluminescenceimagingandeithertreatedthemwith100mg/kgTemozolomide,or30mg/kgPerifosine,oracombinationwith100mg/kgTemozolomideand30mg/kgPerifosine(Temozolomide+Perifosine)for3to5days.ThemicearesacrificedandtumorsanalyzedhistologicallyforcellproliferationbyKi-67immunostaining.Ki-67stainingindexissignificantlyreducedinmicetreatedwitheitherTemozolomide(Ki-67stainingindex=5.5±1.2%,n=4,P=0.0019)orPerifosine(Ki-67stainingindex=3.2±1.1%,n=3,P=0.001)comparedwithControl,demonstratingtheinhibitoryeffectonproliferation.Mostimportantly,thetumorstreatedwithTemozolomide+PerifosinehavethelowestKi-67stainingindex(1.7±1.2%,n=3,P=0.0005).TheadditionaltreatmentwithPerifosineresultsinasignificantlylowerproliferationratethanTemozolomidealone(P=0.0087)[1].Perifosinemarkedlydecreasesp-Aktfrom10minto24hoursandsubsequently,moderatelydecreasedp-S6from1hto24hafterinjection[3].PROTOCOLKinaseAssay[2]Exponentiallygrowingcells(HN12,HN30,andHaCaT)arelysed,and500μgoftotalcellularproteinareusedtoimmunoprecipitateactivecdc2andcdk2complexes.AftercapturingwithgammabindGSepharoseandsubsequentwashes,theactiveimmunecomplexesareassessedforactivityinthepresenceofincreasingconcentrationsofPerifosine(0.1-30μM)orflavopiridol(300nM)inthekinaseassaybuffercontaining[γ-32P]ATP(3000Ci/mmol)and0.2mg/mLhistoneH1,25μMATP.Reactionsareincubatedat37°Cfor30minandterminatedbytheadditionofSDS-gelloadingbuffer,resolvedinSDS,anddried2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEgelsaresubjectedtoautoradiographyandphosphorimaging[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]Cellproliferationstudiesbymeasuringtheuptakeof[3H]thymidineisperformed.Briefly,HNSCCandHaCaTcells(1-2×104/well)aregrownovernightin24-wellplatesandexposedtoeitherPerifosine(0.1-30μM)orPBS(control).Aftertreatment(24-48h),cellsarepulsedwith[3H]thymidine(1μCi/well)for4-6h,fixed(5%trichloroaceticacid),andsolubilized(0.5MNaOH)beforescintillationcounting.Experimentsareperformedintriplicates[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1][3]Drugtreatmentoftumor-bearingmice.Image-positiveEf-lucNtv-amicearetreateddailywithi.p.administrationofbufferaloneasacontrol,ori.p.administrationof100mg/kgTemozolomide,ororaladministrationof30mg/kgPerifosine,oracombinationwithPerifosineandTemozolomidefor3to5days.Themeandosesofthetreatmentsare:Control,5(allfive);Temozolomide,3.75(threetofive);Perifosine,3.75(threetofour);andPerifosine+Temozolomide,3(allthree).Controlbuffersolutionconsistedof5%DMSOand1%Tween80indistilledwater.Rats[3]TofurtherdeterminewhethertheparadoxicaleffectofrapamycinonS6phosphorylationisrelatedtoupstreamsignalsofAkt-mTOR,ratsaretreatedwithPerifosine(20mg/kg,ip,once),anAktinhibitor,30minbeforerapamycinadministration.Ratsaresacrificed1hor6hafterrapamycininjection.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻?SciTranslMed.2018Jul18;10(450).pii:eaaq1093.?IntJBiolSci.2016Mar30;12(5):607-16.?FrontOncol.2021Apr12;11:608570.?JCellBiochem.2020Mar;121(3):2343-2353.?ExpCellRes.2021Jul29;112752.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].MomotaH,etal.Perifosineinhibitsmultiplesignalingpathwaysinglialprogenitorsandcooperateswithtemozolomidetoarrestcellproliferationingliomasinvivo.CancerRes,2005,65(16),