質(zhì)譜藥物發(fā)現(xiàn)

DrugXuehengVivaBiotechLtd,MassspectrometryDrugdiscoveryApplicationofmassspectrometryindrugMassspectrometry–specialSensitivity,sampleprep,Application–specialAffinityselectionmassspectrometryforbiologicalFindingbioactivecompoundsfromnatureusingmassspectrometryComponentsComponentsofmassMassQualitativewhatAccurateQauntitativehowmuchAbsolute–requiringreferencestandard,-LCMS,GCMS,LCMSismostcommonindrugComponentidentificationComponentidentificationfromLCMSMassUVSelectedionCalibrationWhyusemassspectrometry?(advantagesWhyusemassspectrometry?(advantagesoverotheranalyticaltechniques)MuchmorethanVU/Vis,IR,ButlessthanfluorescenceandMuchmorethanVU/Vis,R,fluorescenceandButNMRisprobablymoreMorethanButlessthanUV/Vis,fluorescenceandprobablycomparabletoRandMuchmorethanVU/Vis,R,fluorescenceandButlessthanBetterthananyotheranalyticaltechniquesindealingwithDrugR&D疾病藥物（蛋白）靶點(diǎn)藥物先導(dǎo)化合物先導(dǎo)化合物優(yōu) 臨床前試 臨床試 上市新FromNoesisChemoInformaticsUtilityofMSUtilityofMSinDrug疾病藥物（蛋白）靶點(diǎn)藥物先導(dǎo)化合物先導(dǎo)化合物優(yōu)化臨床前試驗(yàn) 蛋白結(jié)構(gòu)測(cè)定化合物結(jié)構(gòu)測(cè)定蛋白/ MassSpectrometryDependingoncompoundstructureandionization+ + Electrosprayionization 1-10umMassMassSpectrometryM > > MassSpectrometry H++[M [M > >> MassMassMixtureAslongasthereisnomassCouplingwithseparation Canhandleevenmore inthe MassoverlapOKifretentiontime LCMSmost MassspecissensitiveandGoodforyourcompoundofMayalsobebadforyourcompoundofIonIonIonization(especiallyESI)isacompetitiveMoresignalsfromsamplematrixmeanslesssignalfromcompoundsofinterestDealingwithionBettersamplepreparation(toremovematrixCouplingwithbetterseparationtechniqueMassMassSpectrometrySampleSampleprepisasimportantasMSanalysisSometimessampleprepismoreimportantthantheExtractiontechniquesforsampleSolidphaseextractionEasytoProteinprecipitationAffinityselectionmassspectrometryDiscoverhighaffinityligandstowardproteinsfromsolutionmixturesbyaffinityselectionandmassspectrometrydetectionThistechniqueutilizesseveralfeaturesofmass--Mixture--Qualitative:whatarethecompoundsthatbindtothe--Quantitative:howmucharethecompoundsboundtotheproteinvs.freeinsolution(bindingconstant)?RelativeRelativeAffinityScreeningAffinityScreeningSurfacePlasmonResonance(SPR)techniques(BIACore,surface,kineticandbindingconstantinformationThermoFluorrayProtein-compoundco-crystallization–ligandispreferentiallyco-Soaking–liganddiffusesintothecrystalandisretainedthereduetoASMS-AffinitySelection/MassR2&+ 10kDa ASMSASMSisanenrichmentAfactorof10enrichmentoftightbindersvs.non-bindersaftereachroundofselectionQuantitativelyBoundNocompound CompoundwithKd=50%2xlosseachUnbound 10xlosseach CycleMSASMSScreening:SignaltoBeforeAfter3roundsofASMS-ComparisonASMS-ComparisontotraditionalEquilibriumsolutionbindingprocessSimilaritiestotraditionalBinderstoallregionsofproteins(potentiallynovelTargetfunctionneednotbeAnysolubleAnylibraryDiscoveryofaNovelSmall-MoleculeInhibitorforMur-EssentialforbacterialUniquetoprokaryotes ActivityHTSnotNO OSCompoundwithhighaffinitydiscoveredbyOJournalofBiomolecularScreening11(2006)ProteinScience14(2005)3039- bindUDP-MurNAc-tripeptideApplicationApplicationofASMSinnaturalproductDiscovernewmedicinesfromBetterchancesoffindingbioactivecompoundsfromChinesemedicinethansyntheticcompoundsRelativeRelativeChemicalstructureChemicalstructurediversityofnaturalproductsismoresimilartoknowndrugsthansyntheticcompoundMorethan60%knowndrugsarefromderivedfromnaturalproductNatural derivedHowHowtofindbioactivecompoundsfromTraditionaldifferences,etc.Brute-forceisolationandThestructures,etc.DifferentcompoundAfactorof10enrichmentofbindersvs.non-bindersafter QuantitativelyBoundroundof NocompoundCompoundwithKd=50%2xlosseachUnbound10xlosseach1 CycleAdvantagesofAdvantagesofASMSfornaturalproductscreeningIdentificationofligandstospecifictargetsprovidesinformationforOnlynon-covalentinteractionligandsareCandealwithdifferentconcentrationsofNoneedtoseparateandpurifyindividualcomponentsbeforeCanidentifythestructureofCandete