3α-Aminocholestane-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemE3α-AminocholestaneCat.No.:HY-19776CASNo.:2206-20-4分⼦式:C₂₇H₄₉N分⼦量:387.68作⽤靶点:Phosphatase作⽤通路:MetabolicEnzyme/Protease储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验Ethanol:50mg/mL(128.97mM;Needultrasonic)DMSO:<1mg/mL(insolubleorslightlysoluble)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.5794mL12.8972mL25.7945mL5mM0.5159mL2.5794mL5.1589mL10mM0.2579mL1.2897mL2.5794mL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%EtOH>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:≥3.25mg/mL(8.38mM);Clearsolution2.请依序添加每种溶剂：10%EtOH>>90%cornoilSolubility:≥3.25mg/mL(8.38mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性3α-Aminocholestane含有选择性SH2结构域的肌醇-5'-磷酸酶1(SHIP1)的抑制剂，其IC50值约为2.5μM。IC50&TargetIC50:2.5μM(SHIP1)[1]体外研究OPM2cellviabilityiseffectivelyreducedby3α-Aminocholestane(3AC)treatment.RPMI8226andU266cellsshowsignificantlylesssensitivityto3α-AminocholestanetreatmentwhencomparewithOPM2cells,althoughviabilityisdecreasedsignificantlyatconcentrationsof≥12.5μM.Treatmentwith3α-Aminocholestanefor36hseverelyreducesthepercentageofcellsintheSphase,whichisaccompaniedbyanincreaseofcellsintheG2/Mphase.Incontrast,inthelessproliferativeRPMI8226andU266cells,cellcycleprogressionisblockedintheG0/G1phaseupon3α-Aminocholestanetreatment,inconjunctionwithareducedpercentageofcellsundergoingtheSphase[2].体内研究Itisfoundthat3α-Aminocholestane(3AC)resultsinreducedmultiplemyeloma(MM)growthinvivo,asdeterminedbyquantitationoffreehumanIgλlightchainintheplasmaafterOPM2challenge.Inaddition,reducednumbersofcirculatingOPM2cells,asdeterminedbyhumanHLA-ABCstaining,isobservedinperipheralbloodfrom3α-Aminocholestane-treatedmicecomparewithvehiclecontrols.Mostimportantly,3α-Aminocholestanetreatmentresultsinsignificantlyenhancedsurvivalofmiceaftertumorchallenge.In3α-Aminocholestane-treatedmicethatresisttreatment,itisfoundthatMMtumorsexhibitanupregulationofSHIP2,reminiscentofinvitrotreatmentofOPM2cellsandsuggestingthatSHIP1inhibitionmayselectfortumorcellswithincreasedSHIP2expression[2].PROTOCOLCellAssay[2]Cellsaretreatedintriplicateormorewithincreasingconcentrationsofcompounds(including3α-Aminocholestane).CellviabilityisdeterminedwithaCellCountingKitaccordingtothemanufacturer’sinstructions.Theoddsdensity(OD)ofcompound(including3α-Aminocholestane)-treatedcellsisdividedbytheODoftheirvehiclecontrol,andtheviabilityisexpressedasapercentageofuntreatedcells.Resultsareexpressedasmean±standarderrorofthemean(SEM)ofthreeindividualexperiments.Forphosphatidylinositolphosphates(PIP)add-backexperiments,MCF-7cellsaretreatedfor2hwith10μMSHIPinhibitors(including3α-Aminocholestane),afterwhichcellsarewashedandfreshmediumisadded.Cellsaresubsequentlyculturedintheabsence(0μM)orpresence(10or20μM)ofeitherPtdIns(3,4)P2-diC16(P-3416)orPtdIns(3,5)P2-diC16(P-3516)for36h,afterwhichcellviabilityisdeterminedbytheCellCountingKit[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEAnimalNOD/SCID/γcIL2R(NSG)miceareinjectedintraperitoneallywith1×107OPM2cellsand6hlaterreceiveanAdministration[2]initialinjectionof3α-Aminocholestane(3AC)orvehicle.3α-Aminocholestaneissuspendedina0.3%Klucel/H2Osolutionat11.46mMandadministeredbyintraperitonealinjectionof100μLsolution.Vehicle-treatedmicereceive100μLinjectionof0.3%Klucel/H2Osolution.Themicearethentreatedwith3α-Aminocholestaneorvehicledailyforthenext6dandthentwiceperweekintheremaining15wksofthesurvivalstudy[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CancerRes.(2022)82(10):1991-2002.•OxidMedCellLongev.2019Apr28;2019:6527638.•CommunBiol.2022Apr8;5(1):339.•CellBiolInt.2020Dec15.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].ChenZ,etal.SignallingthresholdsandnegativeB-cellselectioninacutelymphoblasticleukaemia.Nature.2015May21;521(7552):357-61.[2].GwennyMFuhler,etal.TherapeuticPotentialofSH2Domain-ContainingInositol-5′-Phosphatase1(SHIP1)andSHIP2Inhibition