.,1,1,Sterilization滅菌,AGeneralDiscussionfromCGMPPerspective從CGMP角度的綜合討論,.,2,2,Outline討論綱要,PARTITerminology專(zhuān)業(yè)用詞的定義GMPRequirementsinTheManufactureofSterilePharmaceuticalProducts無(wú)菌藥品生產(chǎn)過(guò)程中GMP的基本要求PARTIIFacilityDesign生產(chǎn)設(shè)施的設(shè)計(jì)HVACSystem空調(diào)系統(tǒng)EnvironmentalMonitoring(EM)環(huán)境的監(jiān)視PharmaceuticalWater制藥用水Cleaning/Sanitation清潔消毒Personnel無(wú)菌區(qū)操作人員PARTIIIMethodsofsterilization滅菌方法SterileProductionandvalidation無(wú)菌生產(chǎn)和驗(yàn)證TheTrendofSterileProduction無(wú)菌產(chǎn)品生產(chǎn)趨勢(shì),.,3,PresentationOutline概要,USRegulations美國(guó)法規(guī)MoistHeatSterilization濕熱滅菌DryHeat/Depyrogenation干熱/去熱原法SterilizationProcessValidation滅菌工藝驗(yàn)證OtherSterilizationMethods其他滅菌方法,.,4,CodeFederalRegulation美國(guó)聯(lián)邦法規(guī),211.84(c)(3)sterileequipment滅菌設(shè)備Sterileequipmentandasepticsamplingtechniquesshallbeusedwhennecessary必要時(shí)應(yīng)使用滅菌設(shè)備和無(wú)菌取樣技術(shù)。211.94(c)and(d)sterilized(c)Drugproductcontainersandclosuresshallbecleanand,whereindicatedbythenatureofthedrug,sterilizedandprocessedtoremovepyrogenicpropertiestoassurethattheyaresuitablefortheirintendeduse.藥品容器和密封系統(tǒng)應(yīng)清潔并根據(jù)藥品的性質(zhì)和要求，進(jìn)行滅菌，除熱原過(guò)程以確保預(yù)期的用途。(d)Standardsorspecifications,methodsoftesting,and,whereindicated,methodsofcleaning,sterilizing,andprocessingtoremovepyrogenicpropertiesshallbewrittenandfollowedfordrugproductcontainersandclosures.應(yīng)建立并執(zhí)行對(duì)藥品容器和密封系統(tǒng)的規(guī)格或質(zhì)量標(biāo)準(zhǔn)，測(cè)試方法，清潔方法，滅菌和除熱原過(guò)程的相關(guān)書(shū)面程序。,.,5,CodeFederalRegulation美國(guó)聯(lián)邦法規(guī),211.113(a)and(b)sterilization滅菌(a)Appropriatewrittenprocedures,designedtopreventobjectionablemicroorganismsindrugproductsnotrequiredtobesterile,shallbeestablishedandfollowed.應(yīng)該建立并執(zhí)行用于防止非無(wú)菌藥品被致病菌污染的相關(guān)書(shū)面程序。(b)Appropriatewrittenprocedures,designedtopreventmicrobiologicalcontaminationofdrugproductspurportingtobesterile,shallbeestablishedandfollowed.Suchproceduresshallincludevalidationofanysterilizationprocess.應(yīng)該建立并執(zhí)行用于阻止無(wú)菌藥品被致病菌污染的相關(guān)書(shū)面程序。這些程序應(yīng)包括任何無(wú)菌工藝的驗(yàn)證。,.,6,CodeFederalRegulation美國(guó)聯(lián)邦法規(guī),211.167(a)testing（a）測(cè)試Foreachbatchofdrugproductpurportingtobesterileand/orpyrogen-free,thereshallbeappropriatelaboratorytestingtodetermineconformancetosuchrequirements.Thetestproceduresshallbeinwritingandshallbefollowed.對(duì)無(wú)菌和/或無(wú)熱原的每批藥品應(yīng)進(jìn)行相應(yīng)的實(shí)驗(yàn)室測(cè)試以確定其與符合要求。測(cè)試程序應(yīng)有書(shū)面文件并遵照?qǐng)?zhí)行。,.,7,SterilizationMethods滅菌方法,MoistHeat:濕熱Fordrugsanddevices.Themodeofactionisproteindenaturation.用于藥品和設(shè)備。作用方式是使蛋白質(zhì)變性DryHeat:干熱Fordepyrogenationandequipmentsterilization.Themodeofactionisproteindenaturation用于去熱原和設(shè)備滅菌。作用方式是使蛋白質(zhì)變性EthyleneOxide:氧化乙烯Primarilyfordevices.Themodeofactionisalkylationofnucleicacids.主要用于設(shè)備。作用方式是使核酸烷基化Radiation:輻射Primarilyfordevices.ThemodeofactionisDNAstrandbreakage主要用于設(shè)備。作用方式是使DNA鏈破壞Othermethods?其他方法？,.,8,Bioburden生物負(fù)荷,Definition:定義Populationofviablemicroorganismsonorinaproductand/orapackage產(chǎn)品和/包裝上的活性微生物的數(shù)量和類(lèi)型Mixtureoforganisms有機(jī)物混合Indigenousmicroflora地方微植物群Needstobeinactivatedbysterilization需滅菌失活,.,9,Bioburden生物負(fù)荷,BioburdenSources生物負(fù)荷來(lái)源Environment環(huán)境Productcontactsurface,personnel,air產(chǎn)品接觸表面，人員，空氣Materials材料Water,rawmaterials.plastic,paper水，原材料，塑料，紙張CharacteristicsofBioburden生物負(fù)荷特點(diǎn)Typesofmicroorganisms微生物類(lèi)型Resistancetosterilizationprocess對(duì)無(wú)菌工藝的耐受Numberoforganisms有機(jī)物數(shù)量,.,10,BiologicalIndicator(BI)生物指示劑,Microbiologicaltestsystemprovidingadefinedresistancetoaspecificsterilizationprocess微生物測(cè)試系統(tǒng)對(duì)指定滅菌工藝有明確抵抗性。Acharacterizedpreparationofspecificmicroorganismsresistanttoaparticularsterilizationprocess某一確定的微生物(指示劑)應(yīng)具有對(duì)某一特定滅菌工藝的抵抗性,.,11,TypicalBiologicalIndicators典型生物指示劑,Moistheatsterilization濕熱滅菌GeobacillusstearothermophilusBacillusstearothermophilusDryheatandEOsterilization干熱和環(huán)氧乙烷滅菌BacillusatrophaeusBacillussubtilisvar.niger,.,12,FormsofBiologicalIndicators生物指示劑形式,Strips/discsinglassineenvelopes在透明紙信封里的條形板/光盤(pán)Strips/discs條/光盤(pán)Self-contained獨(dú)立包裝的Ampoules安瓶Stripswithmedium中號(hào)條形板Liquidsuspension液體懸浮液Metal金屬Fiberglass玻璃纖維,.,13,ChoiceofanAppropriateBI相關(guān)生物指示劑的選擇,Sterilizationprocess滅菌工藝Cycledesignmethod循環(huán)設(shè)計(jì)方法Productbioburden產(chǎn)品生物負(fù)荷Population數(shù)量Resistance抵抗性,.,14,BIsPreparedbyUser生物指示劑的準(zhǔn)備,Performance性能Resistance抵抗性Population數(shù)量Purity純度D-valueD值Recoverymethods恢復(fù)方法Storagerequirements儲(chǔ)存要求,.,15,BiologicalIndicatorUse生物指示劑使用,PlaceBIwithin把BI放進(jìn)Product產(chǎn)品Package包裝Sterilizerloadtomonitorprocess滅菌器負(fù)荷以監(jiān)視滅菌工藝Exposetosterilizingconditions暴露在滅菌狀態(tài)RemoveBIandtest移除BI和測(cè)試Countsurvivors生存數(shù)量的計(jì)算Growth/nogrowthresponse生長(zhǎng)/無(wú)生長(zhǎng)反應(yīng),.,16,D-valueD值,TheDvalueisthetime,usuallyinminutes,requiredtoachieveinactivationof90%(oronelogarithm)ofapopulationofthetestmicroorganismatspecifiedconditions.D值是在特定條件下微生物數(shù)量降低的90%（或一對(duì)數(shù)）所用的時(shí)間，通常是以分鐘為單位的。BacillusstearothermophilushasaDvalue:嗜熱脂肪芽胞桿菌的D值2minat121oC20minat110oC0.2-0.3minat130oCOfalltheaspectsofsterilizationvalidation,theDvalueisperhapsthemostimportant.ValidatingaprocesswithoutconsiderationoftheDvalueislargelyineffectiveandisnotacceptablefromCGMPperspective滅菌驗(yàn)證中，D值可能是最重要的。不考慮D值的驗(yàn)證過(guò)程多半是無(wú)效的且不被CGMP接受。,.,17,Z-valueZ值,Z-value:numberofdegreesoftemperaturerequiredfora1logarithmchangeintheD-valueZ值：D值1對(duì)數(shù)改變需要的溫度數(shù)Z=-1/slopeofthethermalresistancecurveZ=-1/熱阻力曲線斜率whereSlope=logarithmicchangeinD-value/changeintemperature斜率=D值上對(duì)數(shù)的改變/溫度的改變,.,18,ImpactofZ-valueZ值的影響,Whenz-valueissmall,considerablylessinactivationwillresultbelowreferencetemperatureandconsiderablymoreabovethereferencetemperature當(dāng)Z值較小時(shí)，較低程度的失活將導(dǎo)致溫度低于參考溫度，較大程度的失活將導(dǎo)致溫度高于參考溫度。,.,19,TypicalTemperatureProfiles典型溫度分布,.,20,TypeofSterilization滅菌類(lèi)型,MoistHeatSterilization濕熱滅菌,.,21,MoistHeatSterilization濕熱滅菌,Characteristics:特征Wellunderstoodandwellcharacterizedprocess,firstvalidatedprocessinpharmindustry濕熱滅菌是一已被很好地理解并描述滅菌工藝，也是在制藥工業(yè)中第一個(gè)被驗(yàn)證的滅菌工藝Suitableforawidevarietyofapplications適合于較大應(yīng)用范圍Equipmentisreadilyavailable設(shè)備很容易從市場(chǎng)購(gòu)買(mǎi)到Costonaperusebasisislow每次使用基準(zhǔn)花費(fèi)低,.,22,MoistHeatSterilization濕熱滅菌,Applications:應(yīng)用Terminalsterilizationofparentalproduct注射劑的終端滅菌Sterilizationofequipmentandcomponentsforuseinasepticfilling無(wú)菌灌裝線上設(shè)備和配件的滅菌Sterilizationoflaboratorymaterials實(shí)驗(yàn)室用材料的滅菌In-situsterilizationofprocesspipingandequipment(SIP)工藝管道和設(shè)備在線滅菌,.,23,BasicTypesofMoistHeatSterilization濕熱滅菌基本類(lèi)型,Saturatedsteam飽和蒸汽Autoclaves(self-closing)高壓滅菌柜（半封閉）SIP在線滅菌Superheatedwater過(guò)熱水Spray噴霧Submerged浸沒(méi)的SIP在線滅菌Steam-air-mixture(SAM)水蒸氣空氣混合物,.,24,BasicElementsofSterilizationProcessValidation滅菌工藝驗(yàn)證的基本元素,Emptyvesselheatdistribution空容器熱分布Heatdistributionandpenetration熱分布和滲透3.Microbiologicalchallenges微生物挑戰(zhàn),.,25,SteamSterilizationValidation:Prerequisites蒸汽滅菌驗(yàn)證：前提,OQforanautoclave:高壓滅菌柜運(yùn)行確認(rèn)Emptychambertemperaturemappingwithin1.0oCofthemean空腔體溫度分布圖在平均值的1.0oC內(nèi)Chamberintegritytest(noleaking)腔體完整性測(cè)試CertificationofHEPAfiltrationontheairusedtobreakvacuumorintegritytestingoftheventfilter用于隔斷真空或通氣過(guò)濾器完整性測(cè)試的空氣HEPA過(guò)濾認(rèn)證RequirementsforSIP在線滅菌要求Temperaturemapping溫度分布圖Anintegritytest,whereappropriate相關(guān)完整性測(cè)試UseofBI生物指示劑的使用Allcriticalinstrumentsmustbecalibrated所有關(guān)鍵儀器需校驗(yàn),.,26,SteamSterilizationValidation:Prerequisites蒸汽滅菌驗(yàn)證：前提,Acceptabletestresultsfornon-condensablegases,super-heatedsteamanddrynessshouldbeobtainedforthecleansteamusedfortheautoclave/SIP應(yīng)獲得用于高壓滅菌柜/SIP的潔凈蒸汽中的不凝氣體，過(guò)熱蒸汽及干燥度的可接受測(cè)試結(jié)果5.ToolsfortheconductofthePQstudy:進(jìn)行PQ研究的工具BIwith106sporesandknownDandZvaluesBI有106個(gè)孢子，已知D值和Z值Temperaturesensors溫度傳感器Recordingdevicecapableofsupporting12tempsensorswithanaccuracyof0.5oC,recordingdataeveryminuteorless記錄設(shè)備能支撐12溫度傳感器，精度0.5oC，每分鐘或間隔更短時(shí)間記錄數(shù)據(jù)Meansofintroducingtempsensorsintotheautoclave/SIP將溫度傳感器導(dǎo)入高壓滅菌柜/SIP的方法,.,27,SteamSterilizationValidation:PreparationofPQprotocol蒸汽滅菌驗(yàn)證：PQ方案的準(zhǔn)備,Aprotocolshallbepreparedfor:對(duì)于下列各項(xiàng)應(yīng)建立方案：Newautoclave/SIP新高壓滅菌柜/SIPNewloadingpatternsorproductconfigurations新裝料方式或產(chǎn)品配置Changestoexistingpatterns對(duì)現(xiàn)有裝料方式的變更Changestooperationcycleparameters對(duì)運(yùn)行周期參數(shù)的變更Majorchangetoequipmentasdirectedbychangecontrol變更控制要求的設(shè)備主要變更,.,28,SteamSterilizationValidation:PreparationofPQprotocol蒸汽滅菌驗(yàn)證：PQ方案的準(zhǔn)備,Theprotocolmayinclude:方案可能包括：Objectivesofthevalidationstudy驗(yàn)證研究的目的Identificationanddescriptionofthesterilizeranditsprocesscontrols滅菌器的識(shí)別和說(shuō)明及工藝控制IdentificationofSOPsfortheprocessequipment工藝設(shè)備SOP的識(shí)別DescriptionoforSOPreferenceforinstrumentcalibrationprocedures儀器校驗(yàn)程序的說(shuō)明或SOP參考Identificationofcalibrationproceduresfortemp-monitoringequipment,whichincludeatwopointpre-runcalibrationandapost-runverificationforeachrun溫度監(jiān)測(cè)設(shè)備校驗(yàn)程序的識(shí)別，包括一個(gè)兩點(diǎn)預(yù)運(yùn)行校驗(yàn)和每次運(yùn)行后的確認(rèn)Processparameteracceptancecriteria工藝參數(shù)的驗(yàn)收標(biāo)準(zhǔn),.,29,SteamSterilizationValidation:PreparationofPQprotocol蒸汽滅菌驗(yàn)證：PQ方案的準(zhǔn)備,Adescriptionofthefollowing:以下說(shuō)明Biodurdendeterminationstudies生物負(fù)荷確認(rèn)研究Emptychamberheatdistributionstudies(1oC)空腔體熱分布研究（1oC）Loadedchamber(includingLoadconfiguration,maxladingandminloading)heatpenetrationstudies滿(mǎn)載腔體（包括裝載配置,最大和最少）熱穿透研究Containermappingstudies(maynotneededif100mL)容器分布圖研究（如果容量3logreduction大于3個(gè)對(duì)數(shù)減少值,.,44,AdvantagesofDry-heatDepyrogenation干熱去熱原的優(yōu)點(diǎn),Inactivatespyrogenswhilesterilization滅菌時(shí)使熱原失活Materialsdryattheendofcycle循環(huán)結(jié)束時(shí)物料干燥Corrosiveeffectsareminimal腐蝕性最小Conveyorsystemsallowforhighertempsandshorterdwelltime傳送系統(tǒng)允許更高溫度和更短的停留時(shí)間,.,45,DisadvantagesofDry-heatDepyrogenation干熱去熱原的缺點(diǎn),Slowprocess(airisapoorconductor)降低工藝速度（空氣是不良導(dǎo)體）Heatpenetrationslowerthansteam熱穿透比蒸汽慢Ratevaries(slowforglass,rapidforstainlesssteel)速度各異（玻璃慢，不銹鋼快）Heatmustpenetratetoinnersurfaceviaconduction熱必須通過(guò)傳導(dǎo)穿透到內(nèi)表面Layeringcanoccurduetodifferencesinairdensitywithtemperature;mechanicalcirculationneeded空氣密度，溫度不同可能導(dǎo)致分層，需要機(jī)械循環(huán)Heatdegradationlimitsmaterials熱降解限制材料Contractionduringcoolingmaydrawcontaminants冷卻過(guò)程中接觸可能引起污染,.,46,DepyrogenationbyTunnels通過(guò)烘箱去熱原,HEPA-filtered,verticallaminarairflowinheatingandcoolingzones,orradiantheatersinheatingzoneandverticallaminarairflowincoolingzone在加熱和冷卻區(qū)經(jīng)HEPA過(guò)濾的垂直層流氣流，或加熱區(qū)的輻射加熱器和冷卻區(qū)的垂直層流氣流Conveyorbelttoprovidein-linecontinuousflowofsterileglasswaretoasepticarea傳送帶提供連續(xù)在線的無(wú)菌玻璃器皿到無(wú)菌區(qū)的流動(dòng)Limitedtoonetypeofloadatatime一次限用一種裝載方式Problemtocontrolspeedmatchotherlineequipment(filler)控制速度以匹配其他線設(shè)備（灌裝機(jī)）的問(wèn)題Difficulttoachieveuniformheating,andheatsourcemaygenerateparticles很難達(dá)到均勻加熱，熱原可能產(chǎn)生顆粒Largeproductvolumeneededtojustifytunnel調(diào)整烘箱要求大的產(chǎn)品容量,.,47,ValidationofDepyrogenationCycles去熱原循環(huán)驗(yàn)證,Runengineeringtrialstofindcoldspots進(jìn)行試運(yùn)行以找到冷點(diǎn)Runtrialstodetermineworsecasevial,e.g.,thickestglass,packingeffects試車(chē)以確定最差狀況的玻璃瓶，如最厚的玻璃，緊束效應(yīng)Procureorprepareendotoxinindicators獲得或制備內(nèi)毒素指示劑Run3fullloadswithendotoxinindicatorsinplaceatcoldspotsandrandomsites進(jìn)行3次滿(mǎn)載運(yùn)行，內(nèi)毒素指示劑置于冷點(diǎn)和隨機(jī)位置Acceptancecriteria驗(yàn)收標(biāo)準(zhǔn)min.3logreductioninEUforspikedvials帶孔玻璃瓶3個(gè)對(duì)數(shù)減少值，單位為EU3successfulrunswithcontrolsanddocumentation受控并記錄的3次成功運(yùn)行,.,48,CaseStudy實(shí)例分析DryHeatDepyrogenationTunnel干熱去熱原隧道烘箱,ValidationAcceptanceCriteria驗(yàn)證驗(yàn)收標(biāo)準(zhǔn)Aminimumofthreesuccessfulvalidationrunsmustbeperformed.必須至少進(jìn)行三次成功的驗(yàn)證Distributionthermocoupletemperaturesmustbewithin15oCofthesetpointtemperature,afterstabilization.穩(wěn)定后，分配熱電偶溫度必須在設(shè)定溫度的15oC內(nèi)AminimumcumulativeFHvalueoftwelveminutesmustbedemonstratedforeachpenetrationthermocoupleattheendofeachcycle.每次循環(huán)結(jié)束時(shí)，必須證明每個(gè)滲透熱電偶12分鐘內(nèi)的最少累計(jì)FH值A(chǔ)minimum3-logreductionofendotoxinmustbedemonstratedforeachendotoxinspikedvialexposedtothedepyrogenationcycle.必須證明暴露于去熱原循環(huán)的每個(gè)內(nèi)毒素長(zhǎng)頸瓶至少有3個(gè)內(nèi)毒素對(duì)數(shù)減少值,.,49,CaseStudy實(shí)例分析DryHeatDepyrogenationTunnel干熱去熱原隧道烘箱,ValidationAcceptanceCriteria驗(yàn)證驗(yàn)收標(biāo)準(zhǔn)ThecontrollingRTD(hotzoneEntranceRTD)andthethermocoupleadjacenttothecontrollingRTDmustbewithin10oCofeachotherduringtheexposure/dwellphaseafterstabilization.穩(wěn)定后，暴露/停留階段控制RTD（高溫區(qū)入口RTD）和靠近控制RTD的熱電偶之間溫差必須在10CAminimumof20of26thermocouplesmustbefunctionalandmeetpostcyclecalibrationcheckcriteriaperXXXXXSOP.26個(gè)熱電偶中至少有20個(gè)能起作用，滿(mǎn)足XXXXXSOP后循環(huán)校驗(yàn)檢查標(biāo)準(zhǔn)。,.,50,OtherSterilizationMethods其他滅菌方法,EthyleneOxideSterilization氧化乙烯滅菌Agaseouschemicalagent一種氣態(tài)化學(xué)劑M