Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemECobicistatCat.No.:HY-10493CASNo.:1004316-88-4Synonyms:GS-9350分?式:C??H??N?O?S?分?量:776.02作?靶點(diǎn):CytochromeP450;HIV作?通路:MetabolicEnzyme/Protease;Anti-infection儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:250mg/mL(322.16mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.2886mL6.4431mL12.8863mL5mM0.2577mL1.2886mL2.5773mL10mM0.1289mL0.6443mL1.2886mL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(shí)，請?jiān)?個(gè)?內(nèi)使?，-20°C儲存時(shí)，請?jiān)?個(gè)?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)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(shí)驗(yàn)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(shí)驗(yàn)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:≥2.08mg/mL(2.68mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(2.68mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(2.68mM);ClearsolutionBIOLOGICALACTIVITY?物活性Cobicistat有效，選擇性的細(xì)胞?素酶P4503A(CYP3A)抑制劑，IC50值為30-285nM。Cobicistat?種藥代動?學(xué)增強(qiáng)劑，可增強(qiáng)抗HIV藥物的吸收。IC50&TargetIC50:30-285nM(CytochromeP450)[1]體外研究InHIV-1proteaseenzymaticassayandantiviralcellularassays.CobicistatisinactiveagainstHIV-1protease(IC50>30μM).AndCobicistathasnoinhibitoryeffectagainstHIVreplicationinamulticycle5-dayMT-2HIVinfectionassay(EC50>30μM).InassaysusingMT-2cells,Cobicistatexhibitsminimalcytotoxicity,withaCC50valueabove80μM[1].ThemodeofinhibitionofhumanCYP3AbyCobicistatandRitonavirsharesthesamemechanismofactionfortheinhibitionofCYP3A.ItshowsitsinhibitoryeffectsonCYP3AmayinvolvedirectlyatthehemegroupoftheCYP3Aenzyme[1].TheminimaladverseeffectsofCobicistatintheseassayssuggestalowerpotentialfortoxicityrelatedtoalteredlipidmetabolism.Inthelipidaccumulationassaywiththehumanadipocytes,RitonavirshowsacleareffectwithanEC50of16μM.However,Cobicistatexhibitsnoeffectataconcentrationupto30μM[1].Intheglucoseuptakeassaywithmouseadipocytes,Ritonavirshowsapronouncedeffectattheconcentrationof10μM.Incontrast,theeffectsonglucoseuptakebyCobicistat(10μM)issignificantlyless[1].PROTOCOLKinaseAssay[1]InhibitionofhumancytochromeP450activitiesisdeterminedinduplicateinpooledhumanhepaticmicrosomalfractionsfollowingcurrentscientificandregulatoryguidelines.Reactionconditionsarelinearwithrespecttoincubationtimeandhepaticmicrosomalproteinconcentration.SubstratesarepresentatconcentrationsequaltoorlessthantheirrespectiveKmvaluesdeterminedunderthesamereactionconditions.Metaboliteand/orsubstrateconcentrationsaredeterminedusingspecific,internalstandardcontrolledHPLCMS/MSassays.Forreactionsmonitoringmetaboliteformationthereislessthan20%consumptionofsubstrateduringthereaction.Unlessotherwisenotedmicrosomalfraction,dilutedinpotassiumphosphatebuffer,ispreincubatedwithsubstrateandinhibitorfor5minat37°CandthereactioninitiatedbytheadditionofanNADPHgeneratingsystemfollowedbyfurtherincubationat37°Cwithshaking.Enzyme-selectivepositivecontrolinhibitorsaretestedinparallel.Atappropriatetimesaliquotsofthemixtureareremovedandthereactionterminatedbyadditiontoamixtureofmethanolandacetonitrilecontainingtherespectiveinternalstandard.AftercentrifugationaliquotsofthesupernatantaresubjectedtoHPLC-MS/MSanalysis.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemECellAssay[1]Five-foldserialdilutionsofthetestedcompoundsarepreparedintriplicatein96-wellplates.MT-2cellsareaddedtoplatesatadensityof20,000/wellinafinalassayvolumeof200μL.Aftera5-dayincubationat37°C,thecytotoxiceffectisdeterminedusingacellviabilityassay.OnehundredμLmediaisremovedfromeachwellandreplacedwith100μLofphosphate-bufferedsalinecontaining1.7mg/mLXTTand5μg/mLPMS.Following1-hourincubationat37°C,20μLof2%TritonX-100isaddedtoeachwellandabsorbanceisreadat450nmwithabackgroundsubtractionat650nm.Thedataareplottedascellviabilityvs.drugconcentration.Cellviabilityisexpressedasapercentageofthesignalfromuntreatedsamples(0%cytotoxicity)afterthesubtractionofsignalfromsamplestreatedwith10μMofPodophyllotoxin(100%cytotoxicity).TheCC50valueiscalculatedfromtheinhibitionplotsastheconcentrationofdrugwhichinhibitscellproliferationby50%.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?ActaPharmSinB.2021Mar22.?ActaPharmSinB.2020Aug.?EurJMedChem.2020Oct15;204:112626.?EurJMedChem.2020Aug15;200:112427.?Viruses.2020Apr16;12(4):452.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].LianhongXu,etal.Cobicistat(GS-9350):APotentandSelectiveInhibitorofHumanCYP3AasaNovelPharmacoenhancer.ACSMed.Chem.Lett.,2010,1(5),pp209–213[2].TemesgenZ.Cobicistat,apharmacoenh