AZD3965-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEAZD3965Cat.No.:HY-12750CASNo.:1448671-31-5分?式:C??H??F?N?O?S分?量:515.51作?靶點:MonocarboxylateTransporter作?通路:MembraneTransporter/IonChannel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(193.98mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.9398mL9.6991mL19.3983mL5mM0.3880mL1.9398mL3.8797mL10mM0.1940mL0.9699mL1.9398mL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：0.5%HPMC>>0.2%Tween80Solubility:5mg/mL(9.70mM);Suspendedsolution;Needultrasonic1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(4.03mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.03mM);Clearsolution4.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.03mM);Clearsolution5.請依序添加每種溶劑：20%HP-β-CDinsalineSolubility:5mg/mL(9.70mM);Suspendedsolution;Needultrasonicandwarmingandheatto60°CBIOLOGICALACTIVITY?物活性AZD3965選擇性的MCT1抑制劑，Ki值為1.6nM，?MCT2的選擇性?6倍。IC50&TargetKi:1.6nM(MCT1)[1]體外研究AZD3965isdesignedtoselectivelyinhibitMonocarboxylatetransporter-1(MCT1)andwillthereforebeexpectedtoinfluencethemovementoflactateintoandoutofcells[1].AZD3965treatmentcausesa3.7foldincreaseinintracellularlactateinhypoxicCOR-L103and3.7foldand3.9foldincreasesinnormoxicandhypoxicNCI-H1048cellsrespectively.Inallothercasesa50ofNCI-H1048isincreasedfrom0.14nMto10.5nMinNCI-H1048cells.ThisisconsistentwithAZD3965actingviaMCT1inhibition[2].體內(nèi)研究COR-L103tumorbearingmicearetreatedwith100mg/kgAZD3965orvehicleBIDfor21daysandthetumorvolumemonitored.Pharmacokineticanalysisdemonstratesthat100mg/kgAZD3965BIDresultsinconcentrationsoffreeAZD3965predictedtoinhibitlactatetransport.AZD3965treatmentsignificantlyreducesthegrowthofCOR-L103tumors,althoughtumorregressionisnotseen,consistentwithAZD3965onlytargetingthehypoxicfractionofthetumor[2].PROTOCOLKinaseAssay[1]Cellsareplatedovernightandtreatedwith100nMAZD3965orvehiclefor24hours.Thecellsarethenwashed,onceinPBSandtwicewithlysisbuffer(50mMMops,100mMKCl,5mMMgCl2,1mMEDTA,0.1mMDTT,1mMPMSFsupplementedwith1×minicompleteproteaseinhibitorcocktailtablets.Thecellsareharvestedbyscrapingandcentrifugation,andthepelletsnapfrozenwithoutbufferinliquidnitrogen.Frozenaliquotsofcellsarethawedoniceandre-suspendedinlysisbuffer.Cellsarelysedby3roundsoffreezinginliquidnitrogenandthawingat37°Cfor2minuteseach.Lysatesaresubsequentlycentrifuged(13000g,10min,4°C)untilclearandthenstoredonice.Enzymeactivityinthecelllysatesisdeterminedusingamicro-platereadertomeasureeitherproductionordepletionofNADH/NADPH,throughitsabsorbanceat340/10nm,occurringasaresultofdirectorcoupledenzymereactions.The96wellplatesusedfortheassaysarepre-heatedto37°Cfor10minutespriortostartingreactions,initiatedbytheadditionof5μLcelllysateto95μLofreactionbuffer(50mMMopspH7.4,100mMKCl,5mMfreemagnesium).Thestandardreactionbufferissupplementedtoassaythekineticsofthedifferentenzymes.Absorbancevaluesforcontrolsaresubtractedfromabsorbanceofcorrespondingreactions.Graphpadprism6isusedtoplotV0valuesagainst2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEsubstrateconcentrationanddetermineVmaxandKmvalues.TheVmaxisthennormalisedtotheproteinconcentrationinthecelllysate[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[2]Administration[1]COR-L103xenograftsaregrownbysubcutaneousinjectionof5×106cellsin0.2mLof1:1serum-freeRPMI:Matrigelintothemid-dorsalflankof8to14-week-oldmaleNODscidgammamice.Micearehousedinindividuallyventedcagingsystemsina12-hourlight/12-hourdarkenvironmentandmaintainedatuniformtemperatureandhumidity.Tumorsizeismeasuredtwiceaweekusingcalipersandthevolumecalculatedastumorlength×tumorwidth2/2.30daysafterimplantation,micebearingtumorsbetween150and250mm3arerandomizedintotwogroupsofsixandtreatedwith100mg/kgBIDAZD3965in0.5%hydroxypropylmethylcellulose,0.1%tween80orvehicleonlybyoralgavagefor21days.Measurementsarecontinued3timesaweekforthedurationofdrugtreatmenttoassesstumorgrowthkinetics.Atsacrificetumorsarecollectedtodetermineintra-tumorlactateconcentration.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?Cell.2021Jan21;184(2):370-383.e13.?Cell.2019Jul11;178(2):330-345.e22.?CellMetab.2021Sep17;S1550-4131(21)00421-6.?CellStemCell.2022Apr7;29(4):545-558.e13.?NatMetab.2021Nov11.Seemorecustomervalidat