CCT245737-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemECCT245737Cat.No.:HY-18958CASNo.:1489389-18-5分?式:C??H??F?N?O分?量:379.34作?靶點:CheckpointKinase(Chk)作?通路:CellCycle/DNADamage儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:≥32mg/mL(84.36mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.6362mL13.1808mL26.3616mL5mM0.5272mL2.6362mL5.2723mL10mM0.2636mL1.3181mL2.6362mL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%saline1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:≥2.5mg/mL(6.59mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.59mM);ClearsolutionBIOLOGICALACTIVITY?物活性CCT245737?種可?服的，選擇性的Chk1抑制劑，IC50值為1.3nM。IC50&TargetChk1Chk2ERK8PKD11.3nM(IC50)2440nM(IC50)130nM(IC50)298nM(IC50)RSK2RSK1FLT3MARK3361nM(IC50)362nM(IC50)582nM(IC50)698nM(IC50)NUAK1CLK2BRSK1AMPK711nM(IC50)1370nM(IC50)1660nM(IC50)2970nM(IC50)PHKCDK2/CyclACDK1/CyclB3470nM(IC50)3850nM(IC50)9030nM(IC50)體外研究CCT245737(10μM)shows>80%inhibitionofapanelof124kinases,includingERK8,PKD1,RSK2andRSK1withIC50sof130,298,361and362nM[1].CCT245737abrogatesanVP-16-inducedG2checkpointinHT29,SW620,MiaPaCa-2,andCalu6celllines,withIC50srangingfrom30to220nM[2].體內(nèi)研究CCT245737(150mg/kgp.o.)inhibitstumorgrowthincombinationwithLY188011(100mg/kgiv)inHT29coloncancerxenografts.CCT245737(300mg/kg,p.o.)alsoinhibitstheLY188011(60mg/kgiv)inducedpSer296CHK1autophosphorylationat24hinSW620humancoloncancerxenografts[1].CCT245737(150mg/kg,p.o)alonesignificantlyinhibitstumorgrowthinanEμ-MycmousemodelofhumanB-celllymphocyticleukemia[2].PROTOCOLCellAssay[2]Cytotoxicityisdeterminedasthedrugconcentrationthatgives50%inhibitionoftumorcellproliferation(GI50)usinga96hSulforhodamineB(SRB)assay.InhibitionofintracellularCHK1activityismeasuredusingacellbasedELISAfortheabrogationofanVP-16inducedG2checkpoint(mitosisinductionassay,MIA).TheIC50forG2checkpointabrogation(MIA)isdeterminedinthepresenceofnocodazoleusingUCN01asapositivecontrol.Theactivityindex(AI)isusedasameasureofthecompoundsabilitytoinducemitosisrelativetoitstoxicity(i.e.,ratioofMIAIC50:96hSRBGI50).Routinepotentiationstudiesarecarriedoutusingafixedconcentration(GI50)ofeitherLY188011orSN38incombinationwitharangeofCCT245737concentrationstodeterminethecombinationGI50ofCCT245737.TheabilityofCCT245737toenhanceLY188011orSN38cellkillingisexpressedasapotentiationindex(PI)equaltotheratiooftheGI50forCCT245737aloneversusthecombinationGI50forCCT245737.PIvalues>1indicatepotentiationofthegenotoxicactivity.Inaddition,aseriesofexperimentsiscarriedoutusingfixed,non-orminimallytoxic2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEconcentrationsofCCT245737(≤GI20)witharangeofdifferentconcentrationsofLY188011orSN38todeterminetheextenttowhichCCT245737enhancesdrugcytotoxicitycomparedwiththegenotoxicagentalone,i.e.conventionalPI(ratioGI50genotoxicalone:GI50genotoxiccombinedwithnon-toxicCCT245737concentration,ConPI)[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalHumanHT29colorectalcarcinomacellsareinjecteds.cintotheflanksoffemaleNCrathymicmice6-8Administration[1]weeksofage.Dosingcommenced5daysaftertransplantationwhentumorsreachameandiameterof5.5mm.LY188011(100mg/kgi.v.)isdosedinsalineondays0,7and14andcompounds4(CCT245737)and41(150mg/kgp.o.)in10%DMSO20%PEG400,5%Tween80,65%water,24and48haftereachdoseofLY188011.Tumorsaremeasuredandbodyweightsrecordedthreetimesweekly.Animalsareculledonanindividualbasiswhentumorsreachapredeterminedhumaneendpoint(meandiameter[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].OsborneJD,etal.MultiparameterLeadOptimizationtoGiveanOralCheckpointKinase1(CHK1)InhibitorClinicalCandidate:(R)-5-((4-((Morpholin-2-ylmethyl)amino)-5-(trifluoromethyl)pyridin-2-yl)amino)pyrazine-2-carbonitrile(CCT245737).JMedChem.201[2].WaltonMI,etal.TheclinicaldevelopmentcandidateCCT245737isanorallyactiveCHK1inhibitorwithpreclinicalactivityinRASmutan