Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEFMK9aCat.No.:HY-100522CASNo.:1955550-51-2分?式:C??H??FN?O?分?量:392.42作?靶點(diǎn):Autophagy作?通路:Autophagy儲(chǔ)存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:100mg/mL(254.83mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.5483mL12.7414mL25.4829mL5mM0.5097mL2.5483mL5.0966mL10mM0.2548mL1.2741mL2.5483mL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；?旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?，-20°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?。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(guò)程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過(guò)加熱和/或超聲的?式助溶)1.請(qǐng)依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(6.37mM);Clearsolution1/2MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(6.37mM);Clearsolution3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.37mM);ClearsolutionBIOLOGICALACTIVITY?物活性FMK9aautophagin-1的抑制劑，在FRET和LRA試驗(yàn)中測(cè)定的IC50值分別為80和73nM。IC50&TargetIC50:80nM(autophagin-1,FRETassay);73nM(autophagin-1,LRAassay)[1]體外研究ATG4Borautophagin-1isacysteineproteasethatcleavesATG8familyproteins.ATG4Bplaysessentialrolesintheautophagosomeformationandtheautophagypathway.FMK9ashowsstronginhibitionofATG4B,withIC50valuesof80and73nMintheTR-FRETandcellular-basedLRAassays,respectively.LC?MS/MSstudyindicatesthatFMK9aformsanirreversiblecovalentbondwiththemorereactivethiolgroupofCys74locatedatthecatalyticsiteofATG4BandthusinactivatesATG4Bproteolyticactivity[1].體內(nèi)研究FMK9ashowsmoderatesolubility(LYSA:41μg/mL)andhighhumanandmouselivermicrosomeclearancesof13.9and70mL/kgperminute[1].PROTOCOLKinaseAssay[1]FMK9aisdissolvedinDMSO.5μLofserial-dilutedFMK9a(1%DMSO)inassaybufferisaddedinto5μLofpurifiedATG4B(final0.1nM)in384-wellplate,andthesolutionisincubatedatroomtemperature(rt)for30min.Then,5μLofHis-GATE-16-GST(final20nM)isaddedintothewells,andthesolutionisincubatedforanother30min.5μLofdetectionsolution(final2nMofEu-anti-Hisand20nMofUlight-anti-GST)isadded,andtheresultingmixtureisincubatedatrtfor40min[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?Autophagy.2022Mar8;1-18.?ResearchSquarePreprint.2021Jul.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].QiuZ,etal.DiscoveryofFluoromethylketone-BasedPeptidomimeticsasCovalentATG4B(Autophagin-1)Inhibitors.ACSMedChemLett.2016Jun25;7(8):802-6.McePdfHeight2/2MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemECaution:Producthasnotbeenfullyvalidatedform