Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemEZ-VAD(OMe)-FMKCat.No.:HY-16658CASNo.:187389-52-2分?式:C??H??FN?O?分?量:467.49Sequence:Z-Val-Ala-Asp(OMe)-FMKSequenceZVA-D(OMe)-FMKShortening:作?靶點(diǎn):Caspase作?通路:Apoptosis儲(chǔ)存?式:Powder-80°C2years-20°C1yearInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:100mg/mL(213.91mM;Needultrasonic)掃描?維碼，運(yùn)?溶解?案計(jì)算器獲得適合您實(shí)驗(yàn)體系的溶解?案MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.1391mL10.6954mL21.3908mL5mM0.4278mL2.1391mL4.2782mL10mM0.2139mL1.0695mL2.1391mL請(qǐng)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(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存?式和期限。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請(qǐng)先按照InVitro?式配制澄的儲(chǔ)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(chǔ)備液可以根據(jù)儲(chǔ)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(qǐng)依序添加每種溶劑：1%DMSO99%saline1/4www.MedChemEwww.MedChemE2.Solubility:≥0.52mg/mL(1.11mM);Clearsolution請(qǐng)依序添加每種溶劑：10%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.5mg/mL(5.35mM);Clearsolution此?案可獲得≥2.5mg/mL(5.35mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲(chǔ)備液加到400μLPEG300中，混合均勻；向上述3.體系中加?50μLTween-80，混合均勻；然后繼續(xù)加?450μL?理鹽?定容?1mL。請(qǐng)依序添加每種溶劑：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(5.35mM);Clearsolution此?案可獲得≥2.5mg/mL(5.35mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲(chǔ)備液加到900μL20%的SBE-β-CD?理鹽??溶4.液中，混合均勻。請(qǐng)依序添加每種溶劑：10%DMSO90%cornoilSolubility:≥2.5mg/mL(5.35mM);Clearsolution此?案可獲得≥2.5mg/mL(5.35mM，飽和度未知)的澄溶液，此?案不適?于實(shí)驗(yàn)周期在半個(gè)?以上的實(shí)驗(yàn)。5.以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲(chǔ)備液加到900μL??油中，混合均勻。請(qǐng)依序添加每種溶劑：5%DMSO40%PEG3005%Tween-8050%saline6.Solubility:≥2.62mg/mL(5.60mM);Clearsolution請(qǐng)依序添加每種溶劑：5%DMSO95%(20%SBE-β-CDinsaline)Solubility:≥2.62mg/mL(5.60mM);ClearsolutionBIOLOGICALACTIVITY?物活性Z-VAD(OMe)-FMK(Z-Val-Ala-Asp(OMe)-FMK)?種不可逆的pan-caspase抑制劑。Z-VAD(OMe)-FMK泛素C末端?解酶L1(UCHL1)抑制劑。Z-VAD(OMe)-FMK通過靶向UCHL1活性位點(diǎn)對(duì)UCHL1進(jìn)?不可逆地修飾[2]。IC50&TargetCaspase體外研究Z-VAD(OMe)-FMK(Z-Val-Ala-Asp(OMe)-FMK)isabroad-spectrumcaspaseinhibitor,hasbeenshowntoinhibittheintracellularactivationofcaspase-likeproteases.TheinjectionofZ-VAD(OMe)-FMKsuppressesthecaspase-3activityinlungtissues,andsignificantlydecreasesthenumberofterminaldUTPnick-endlabeling-positivecells[1].Z-VAD(OMe)-FMKeffectivelyinhibitsUCHL1'sreactionwithhemagglutinin-taggedubiquitinvinylmethylester(HA-UbVME)attheconcentrationof100μM[2].Z-VAD(OMe)-FMKisadministeredintraperitoneallyat1hourbeforeand6hoursafterSAH.Expressionofcaspase-3andpositiveTUNELisexaminedasmarkersforapoptosis.Z-VAD(OMe)-FMKsuppressesTUNELandcaspase-3staininginendothelialcells,decreasescaspase-3activation,reducesBBBpermeability,relievesvasospasm,abolishesbrainedema,andimprovesneurologicaloutcome[3].Z-VAD(OMe)-FMKisacell-permeablecaspaseinhibitor,efficientlyblockscelldeathinducedbySMNdeficiency[4].體內(nèi)研究ThesurvivalrateofmiceisprolongedsignificantlybytheinjectionofZ-VAD(OMe)-FMK(Z-Val-Ala-Asp(OMe)-FMK).AllmicesuccumbedtoLPSwithin30hours.Bycontrast,themicetreatedwithZ-VAD(OMe)-FMKsurvivesignificantlylongerand27%ofthemicesurvivedmorethan7days[1].2/4www.MedChemEwww.MedChemEPROTOCOLCellAssay[4]PCRproductscontainingcodingsequencesforthedSMN(forwardprimer:5′-TAATACGACTCACTATAGGGAAGACGTACGACGAGTCG-3′;andreverseprimer:5′-TAATACGACTCACTATAGGGGTGGTGCTGGCTTCTTTC-3′;productlength,601bps;boldanditalicslettersrepresentT7promotersequences)andcontrolDrosophilaPresenilin(dPsn)gene(forwardprimer:5′-TAATACGACTCACTATAGGGTGGCTGCTGTCAATCTC-3′;andreverseprimer:5′-TAATACGACTCACTATAGGGCGATAGCAACGCTTCTTG-3′;productlength:543bps)areobtainedandgel-purified.Double-strandedRNAs(dsRNA)aregeneratedbytranscriptionwithRibomaxT7TranscriptionkitanddigestedwithRnase-freeDNase.ThedsRNAproductsareethanolprecipitatedandannealedbyincubationat65°Cfor30minandthenslowlyallowedtocoolatroomtemperature.TheannealeddsRNAproductsareanalyzedona1%agaorsegeltoensurethemajorityofdsRNAexistedasasingleband.ThedsRNA(2μg)and/orplasmidDNAs(2μg)areintroducedintocellsbyusingCellfectin.Caspaseinhibitionisachievedbyusing50μMofZ-VAD(OMe)-FMKintheculturemedium[4].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1][3]Miceusedinthisstudyare5-to6-week-old(20to22g)ICRmales.Miceareinjectedwith30mg/kgLPSfromE.coliserotypeO111:B4throughthetailvein.AsingleintravenousinjectionofZ-VAD(OMe)-FMK(0.25mg)ismade15minutesbeforeLPSinjection,followedbythreeintravenousinjectionsofZ-VAD(OMe)-FMK(0.1mgeach)perhour.Controlmiceareinjectedwiththesamevolumeof1%DMSOinsterilesaline.Rats[3]MaleSprague-Dawleyratsweighing300to350gareanesthetizedwithα-chloralose(40mg/kgIP)andurethane(400mg/kgIP).Animalsareintubated,andrespirationismaintainedwithasmallanimalrespirator.Rectaltemperatureismaintainedat37±0.5°Cwithaheatingpad.Theleftexternalcarotidarteryisisolatedanda4.0monofilamentnylonsutureisinsertedthroughtheinternalcarotidarterytoperforatethemiddlecerebralartery.SAHisconfirmedatautopsyineachrat.Sham-operatedratsunderwentthesameproceduresexceptthatthesutureiswithdrawnafterresistanceisfelt.Z-VAD(OMe)-FMK(50μMper0.3mL)isinjectedintraperitoneallyat1hourbeforeand6hoursafterSAHinduction.Invehiclegroup,ratsunderwentSAHinductionandaretreatedwiththesamevolumeofvehicle(DMSOdilutedinphysiologicalbuffersolution).Notreatmentisappliedinsham-operatedanimals.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?Nature.2020Apr;580(7803):386-390.?Cell.2020Mar5;180(5):941-955.e20.?Cell.2018Sep6;174(6):1477-1491.e19.?CellMetab.2021Feb2;33(2):424-436.e10.?CellRes.2018Dec;28(12):1171-1185.Seemorecustomervalidationsonwww.MedChemEREFERENCES3/4www.MedChemEwww.MedChemE[1].KawasakiM,etal.Protectionfromlethalapoptosisinlipopolysaccharide-inducedacutelunginjuryinmicebyacaspaseinhibitor.AmJPathol.2000Aug;157(2):597-603.[2].ParkS,etal.Neurovascular