AM966-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEAM966Cat.No.:HY-15277CASNo.:1228690-19-4分?式:C??H??ClN?O?分?量:490.93作?靶點:LPLReceptor作?通路:GPCR/GProtein儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:100mg/mL(203.70mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.0370mL10.1848mL20.3695mL5mM0.4074mL2.0370mL4.0739mL10mM0.2037mL1.0185mL2.0370mL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)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：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:2.5mg/mL(5.09mM);Suspendedsolution;Needultrasonic1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(5.09mM);ClearsolutionBIOLOGICALACTIVITY?物活性AM966?種?親和性的選擇性LPA1-拮抗劑，抑制LPA-刺激的細(xì)胞內(nèi)鈣釋放，IC50為17nM。IC50&TargetLPA1[1]體外研究AM966isapotent,selective,orallybioavailableLPA1receptorantagonist.AM966inhibitsLPA1-mediatedchemotaxisofhumanA2058melanomacells(IC50=138±43nM),IMR-90humanlungfibroblasts(IC50=182±86nM)andCHOmLPA1cells(IC50=469±54nM)[1].LPA-inducedERK1/2activationiscompletelyblockedbyAM966(100nM),whichselectivelyantagonizesLPA1overLPA2-5,withanIC50valueof3.8±0.4nM.Pre-treatmentwithAM966(100nM)completelyblocksERK1/2phosphorylationinducedbyeitheramitriptylineormianserin[2].體內(nèi)研究AM966(30mg/kg,BID)reducesvascularleakage,inflammationandlunginjuryandinflammationina3daybleomycinmodel.AM966inhibitslungfibrosis,maintainsmousebodyweightanddecreaseslunginflammation14daysafterbleomycinlunginjury.AM966reducesvascularleakage,tissueinjuryandpro-fibroticcytokineproductioninthe14daybleomycinstudy.AM966demonstratesgreaterefficacycomparedtopirfenidoneinthe14daybleomycinmodel.AM966decreasesmortalityandfibrosisatlatetimepointsafterbleomycininjury[1].PROTOCOLCellAssay[2]CHO-K1cellsaregrownto80%confluencyin12-wellplates,serum-starvedfor24handincubatedinserum-freemediumwithAM966.After21h,[3H]thymidine(0.5μCi/well)isaddedandtheincubationiscontinuedfor3h.Themediumisthenremoved,andthecellsareplacedoniceandwashedtwicewith1mLofice-coldPBScontaining5%trichloroaceticacid.Cellsaresolubilizedand[3H]thymidineincorporationisdeterminedbyliquidscintillationcounting.Assaysareperformedintriplicate[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]TheoralexposureofAM966isdeterminedinfastedmice.AnimalsreceivedAM966(10mg/kg)invehicle(water)byoralgavageandarethenkilledbyCO2inhalationat1,2,4,8and24hpostdose(n=2animalspertimepointforeachtestcompound).Blood(approximately300μL)iscollectedviacardiacpunctureintoEDTA-containingtubesandcentrifugedat1450×gfor10min.TheplasmaisremovedandanalysedforAM966contentbyliquidchromatography-massspectrometry(LCMS).Briefly,knownamountsofAM966areaddedtothawedmouseplasmatoyieldaconcentrationrangefrom0.8to4000ng/mL.Mouseplasmasamplesareprecipitatedusingacetonitrile(1:4,v:v)containingtheinternalstandardbuspirone.A10μLaliquotoftheanalytemixtureisinjectedusingaLeapPALautosampler.AnalysesareperformedusinganAgilentZorbaxSB-C8column(2.1×50mm;5μm)linkedtoaShimadzuLC-10ADVPwithSCL-10AVP2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEsystemcontroller.TandemmassspectrometricdetectioniscarriedoutonaPESciexAPI3200inthepositiveionmode(ESI)bymultiplereactionmonitoring.Thecalibrationcurvesareconstructedbyplottingthepeak-arearatioofanalysedpeaksagainstknownconcentrations.Thelowerlimitofquantitationis0.8ng/mL.Thedataaresubjectedtolinearregressionanalysiswith1/x2weighting.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?CellMetab.2022Mar10;S1550-4131(22)00083-3.?Autophagy.2022Feb27;1-22.?NeuropsychopharmacolRep.2019Sep;39(3):156-163.?BiochemPharmacol.2015Jun15;95(4):311-23.?Apoptosis.2019Jun;24(5-6):478-498.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].Swaney,JS,etal.Anovel,orallyactiveLPA1receptorantagonistinhibitslungfibrosisinthemousebleomycinmodel.BrJPharmacol.2010Aug;160(7):1699-713.[2].OlianasMC,etal.AntidepressantsactivatethelysophosphatidicacidreceptorLPA(1)toinduceinsulin-likegrowthfactor-Ireceptortransactivation,stimulationofERK1/2signalingandcellproliferationinCHO