冻干显微镜下的冻干工艺 英国凯文.沃德博士.pptx

冻干显微镜下的冻干工艺 英国凯文.沃德博士.pptx

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FormulationDesignandCharacterisationforSuccessfulFreeze-DryingCycleDevelopment,冻干显微镜下的冻干工艺Dr.KevinWard,MRSC凯文.沃德博士DirectorofR&D,BiopharmaTechnologyLtd.,WinchesterUK英国Biopharma技术有限责任公司研发总监ChairofPharmaceutical&HealthcareSciencesSocietyFreeze-DryingSpecialInterestGroup,SynopsisofPresentation,2,SomegeneralrulesforformulationdevelopmentandafewverybasicrulesforcycledevelopmentMethodsofformulationcharacterisationFreeze-DryingMicroscopy（FDM）DTA/electricalimpedanceanalysis（Zsin）Residualmoistureanalysisinthelyophilisedproduct,TheIdealFormulation?

3,Insomerespects,theactiveingredient/materialaloneisthebestformulation:

Lowestsolutedensity（therefore,lowerresistancetovapourflow）Noexcipient（in）compatibilityissuesLowercostofmanufactureHowever,theactiveingredientmayneedtobestabilisedpriortoandduringfreeze-drying,FormulationIssues

（1）,4,Itshouldberememberedthat:

FREEZINGinvolvesCONCENTRATION“Freezingisinitselfaformofdehydration”（FelixFranks）DryingmayrisktheremovalofwaterinvolvedinmaintainingthestructureoftheAPI（especiallyforproteins）,FormulationIssues

（2）,5,Lowvolumeofhigherconcentrationshouldfreeze-dryfasterthanlargervolumeofalowerconcentrationManyproteinsloseactivitywhenfreeze-driedfromlowconcentrationsHowever,somematerials（especiallyorganisms）aredifficultorimpossibletoconcentratethemwithoutdamage,FormulationIssues（3）,6,Additionally,someproductsarenotstableinsolution,therebyrequiringpHbufferingand/orotherstabilisationevenbeforefreeze-dryingstartsHowever,rememberthatpHbuffersaredesignedtoworkinsolutiontherearenoguaranteesforthefrozenordriedstate!

APIcharacteristics,7,Crystallineoramorphous?

Teu/Tg/TcBulkcharacteristicswhenfreeze-driedSolubilityConcentrationrequiredpriortoFDpH-stabilityplotIEP&aggregationissuesforproteins,WhatareweformulatingtopreventfortheAPI?

8,DestabilisationinliquidstateDamagebythefreezingprocessLossofactivityduringdryingDegradationduringstorageTherefore,needexcipientsthatarechemicallycompatiblewiththeAPIduringalltheabovestages,Formulationforfreeze-dryinginvolvestheuseofexcipientsto,9,providemechanicalstrength（bulk）affordthermalstability（ahighTcritical）duringlyophilisationandinthedriedproductprotecttheactiveingredient（s）fromdamagebefore,duringandafterprocessinggivecorrectpH,andtonicitywhererequired（sometimesachievedbyreconstitutingmediumratherthanstartingsolution）,CommonExcipients:

pros&cons,10,*fulfilthebasicrequirementofremainingamorphousbutprotectiveabilitydependsonAPI*PEGoftenprovidescryoprotectionbutnotnecessarilylyoprotectionasitcancrystallise,“Lyo-friendly”buffers,11,CitrateTrisGlycine/HistidinePhosphateoftenbestavoidedduetopHshiftsonfreezing,resultingfromdi-sodiumsaltcrystallisingoutOtherbufferssuchasacetate,HEPES,borate,phthalate,arelesswellstudiedforfreeze-dryingbutmaybesuitable,Otherissuesinformulatingforfreeze-drying,12,Mixingamorphousand“crystallising”componentstogether:

Phaseseparation（ice+glass+crystals）Unpredictable“criticaltemperature”Possiblemicrocollapse/micromeltingInhibitionofcrystallisationResultingmetastablecomponentscouldchangeovertimeindrystate,“Extrascientific”issuesaffectingexcipientselection,13,EthicalacceptabilityintargetmarketPreviousacceptancebyregulatorybodies（FDA,MHRAetc.）foreachmodeofuse（e.g.in-vitro,PO,SC,ID,IP,IM,IV）GradeofpurityavailableCostSupplychainreliability,Vialsoffreeze-driedproduct,GoodOK,Poor,Poor,Theproductinthe“Poor”vialshasbecomesoftanddenseduringfreeze-drying,becauseithasbecomewarmerthanits“CriticalTemperature”!

14,“WhatistheCriticalTemperatureforourproduct?

”,15,The“CriticalTemperature”willbe:

Theeutectictemperature（Teu）forcrystallinematerialsThecollapsetemperature（Tc）foramorphousmaterials（somewhereatorabovetheglasstransitiontemperature）Theloweroftheabovetemperaturesformixedsystems（dependingonwhethermicro-collapseisacceptable）,Wecananalysethecriticaltemperatureofaformulationbeforefreeze-dryingit,forexampleusing:

Freeze-DryingMicroscopy（FDM）Impedance（Zsin）andThermalAnalysis,Freeze-dryingmicroscopy（FDM）,FDMisthestudyoffreeze-dryingatthemicroscopiclevelFDMallowsdeterminationofcollapse,meltingand“qualitativephenomena”suchasskinformation,16,WhatisaFreeze-DryingMicroscope?

Effectivelyamicrofreeze-dryerwherethefreeze-dryingofasmallsamplemaybeobservedFirstdesignsinthemid-1960sNowmanufacturedcommercially,17,SamplePreparationforFDM,Sampleholder,Block,Sampleloadingtakesabout60seconds.Routineanalysisusuallytakes3090minutes,SideDoo18r,SampleFormatinLyostat2,Temperature-ControlledBlock,LightSource（frombelow）,Aperture,Quartzcoverslip（16mmdia.）,2lofsample,ObjectiveLens（usually10x）Glasscoverslip（13mmdia.）MetalSpacer（70mthick）,19,IdeallytherawformulationisusedSometimesnecessarytousesamplesthathavepreviouslybeenfrozenorlyophilisedAfterloadingthesample,theLyostat2issettocooltothedesiredtemperature,Thesampleisallowedtocoolandfreeze（Note:

foreutecticmaterials,therewillbemorethanonefreezingevent!

）,SampleLoadingandCooling,20,Whensamplereachestheholdingtemperatureandhasbeenobservedtofreeze,vacuumpumpisswitchedonanddryingbegins.,Sublimationinterfacecanbeseenmovingthroughthefrozensample.,Frozensample,Driedsample,Sublimationfront,On-lineplot,Temp/timetable,INITIALFDMIMAGE,21,Increasingordecreasingthetemperatureofthesampleallowsyoutoviewitsfreeze-dryingcharacteristics.,Byexaminingthefreeze-driedstructurebehindtheinterface,thecollapsetemperatureofthematerialcanbedetermined.,ThetemperaturemaybecycledinordertoevaluateTcmoreclosely,Frozensample,Sublimationfront,Collapsedmaterial,INTERPRETATIONOFEVENTS,22,Samplestructurelostwhencollapsetemperaturewasexceeded.,Structureregainedassamplewasre-cooledtobelowitscollapsetemperature.,Frozensample,Collapsedsample,Regainedstructure,Sublimationfront,INTERPRETATIONOFEVENTS,23,100%structurehasbeenregainedbyloweringthesampletemperature.,Sampletemperaturewasagainincreasedtoaboveitscollapsetemperature,causingthesampletocollapse.,Driedsamplewithstructure,Collapsingagainonreheating,Frozensample,Sublimationfront,INTERPRETATIONOFEVENTS,24,Micro-collapse（seee.g.Wang,2004）,BelowTcofamorphousphase,AboveTcofamorphousphase,Asimilareffectmayalsobeobservedduetothemeltingofcrystallinecomponent（s）ontoarigidamorphousstructure（dependingonwhichhasthelowercriticaltemperature）,Macroscopicallysimilarbutisit:

Wetter?

Lessstable?

Moredifficulttoreconstitute?

25,FDMimageofanaqueoussolutionof2%Mannitol+1%Glucose,Frozenmaterial（Dryingfront）,Regionswithgooddriedstructure.Justmannitol?

Regionsof（micro）collapse.Justglucose?

-41oC,aroundTcforglucose.Possibleevidenceofvisiblemicro-collapse26,So,whatelsecanFDMtellus?

27,Eutecticmeltingtemperature,NaClBelowEutecticTemperature,Frozen,28,Dry,NaClAboveEutecticTemperature,Notechangesinappearanceoffrozenstructure,Eutecticliquid,29,So,whatelsecanFDMtellus?

30,EutecticmeltingtemperatureMaygivesomeindicationofskin（crust）formationpotentialofaformulation,Layerofconcentratedsoluteatedgeofsample,Crustformation

（1）,31,CrustFormation

（2）,Dryingonlyoccursthroughbreaksinthecrust,32,So,whatelsecanFDMtellus?

33,EutecticmeltingtemperatureMaygivesomeindicationofskin（crust）formationpotentialofaformulationWhetherheat-annealingmaybeofbenefitToincreaseicecrystalsizeandwhatconditionsarerequiredforthis（aboveTg?

）Toencouragesomecomponentstocrystallise,Effectofannealingonicecrystalsize,Samplecooledto-40C,thenwarmedto-10C,Samesampleafterafurther15minutesat-10C,Experimentscanbecarriedouttocompareratesofchangeatdifferenttemperatures,inordertoestablishwhatannealingtemperaturemightb,mostefficienttouseinthefreeze-dryer.,34,SamplePolariser,FDMsetupwithpolarisedlightCameraAnalyser,35,Effectofannealingonsolutebehaviour:

FDMwithpolarisedlightfunction,Samplequenchcooledbelow-40CNosignofcrystals（nolightrotation）,SameSamplenowdryingat-18CPolarisershowspresenceofcrystals（whiteareas）,36,FurtherapplicationsofFDM,37,Itispossibletoexaminedifferencesinrelativedryingrates:

FordifferentformulationsForaspecificformulationatdifferenttemperaturesRef:

Zhai,S.,Taylor,R.,Sanches,R.andN.K.H.Slater（2003）.MeasurementofLyophilisationprimarydryingratesbyfreeze-dryingmicroscopy.Chem.Eng.Sci.58,2313-2323,DTAandElectricalImpedanceanalysis（Zsin）ofFrozenFormulations,38,DifferentialThermalAnalysis（DTA）,39,EffectiveyetsimpleandinexpensivemethodofanalysingfrozensolutionsGivesexothermicandendothermicevents,whichcanindicate:

Glasstransitions（amorphous）Eutecticmelts（crystalline）Crystallisations（amorphoustocrystalline）AtBiopharma,weusethisincombinationwithelectricalimpedance（Zsin）analysistogiveamorecompletepicture,ElectricalImpedance（Zsin）Analysis,40,Thisisamoresophisticatedversionofelectricalresistance（R）analysisImpedance（Z）isacombinationofResistance+Inductance+CapacitanceLookingatZ（ormorespecificallyZsin）cangivemoredetailedinformationaboutfrozensolutemobility（Rey,1999）.,InvestigatingZsinMethods

（1）,Wehavedevelopedadevice（Lyotherm2）incollaborationwithProf.LouisRey,whichiscapableofanalysingImpedanceatafrequencyof1000Hz,Lyotherm2allowsbothDTAandImpedance（Zsin）analysistobecarriedoutonasampleinthefrozenstatelargesamplevolumetogivestrongersignal,41,InvestigatingZsinMethods

（2）,Appr