_ The Science and
`Practice of Pharmacy
`
`Bemmgton
`
`21st E DIT I O N
`
`Novartis Exhibit 2024.001
`
`Novartis Exhibit 2024.001
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`·! ,.,·:Edit
`Managing Editor: Matthew J, ffauber
`Marketing Manager: Marisa A. O'Brien
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`material contai:ned herein. This publication contains information relating to general principhlslf medical <:are which should not
`be construed as specific instructions for individual patients. Manufacturers productinformatida,nd package inserts should be
`reviewed for current information, including contraindications., dosages and precautions,
`
`·»··.,«,
`
`Printed in the United States of Amel'ica
`Entered according to Act of Congress, in the year 1885 by Joseph P lkmington, in ;}e ·6mi!tof' Librarian of Congress. at
`J
`·
`Washington DC
`. ,.,
`I
`!
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`,(
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`Copyright 1889, 1894, 1905, 1907, 1917, by ,Joseph P lkmington
`
`Copyright J:926, 1936., by th.e Joseph P Rentington Estate
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`Copyright 1948, 1,951, by the Philadelphia College of Pharmacy and Science
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`Copyright 1956, 1960, 1965, 1970, 1975, 1980, 1985, 1990, 1995, by the Philadelphia Oollege of Pjrmacy and Science
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`Copyright 2000, 2006, by the University of the Scfonces in Philadelphia
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`All Rights Reserved
`Libra~y of Congress Catalog Card lnfonnation is ayailabie
`JSBN 0-7817-4673-6
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`The publishers have made e,uiry .effort to trace the copyright holdr,rs for born>wed materiµl: ll fliey
`' any, they will be pleased to make the necessary arrangements at the fi.rst .opportnnity.
`Th~ use of structural fonnulas from USA.N and the USP Diel.ion.a,ry of Drug Names i11 b;_y per.
`Conuentiords n<,t r.esponsi.hle for any inaccuracy contained herein.
`
`'J:his text is not intended ta rcipresent, nor shall it be interpreted to be, the equivakap ,
`, ,, :~d States Pharmacopeia (USP) and I or the Nat ion.al Formulary (NP). ln tfu! €!Jlfiit~f~1~ .
`cun,wit offieidl USP or NF st.and.ards of strength, quality, purit:,,\ packaging and labei ·
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`hereiW/ t;~e conte:rJ and effect of the o{!icial mmpewiilt sluill prevail.
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`To J}'utehase additional copies of this bonk call our t.'tk~tomer sen'lce depfJ.rtment at (
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`dverte.n.tly overlooked
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`USP C:orwen.tion. The
`
`' tte for the offkial
`i.ficrep<tnliy between the
`· esentations of them
`
`ers to (301)
`
`12345678910
`
`Novartis Exhibit 2024.002
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`used in pharmaceutical technology and the equipment em(cid:173)
`ployed to carry out these methods:
`
`Method
`Moist heat steriliwt:ion
`
`Dry heat sterili1eation
`
`Chemic,,il cokl .~te:rilir.ation
`
`Rudiation ~t.erilization
`
`"J<'iltration
`
`Equipment
`Satnrnt-<!d steam m.1toclave,
`Sttpe~heat.ed wawr at1t-Oda,·e~
`Air OV'er ~team autodave.s
`Bat.ch ster!.lir.ers
`Contim,ous tunnel sterilizers
`Ethylctie oxide
`Vaporized hydrogen )WToxide
`Bydrogm _pcroxidc/steum
`Ot.h.crgase.
`Eledromagnetic
`Particulate
`:t1fombm,ies
`
`DEFINITIONS
`The following terms, relating to sterilization, shou]d be under•
`stood by those can·ying out storilization processes or handling
`stel'ile products:
`Antisepti~~A substance that arrests or prevent,; tl:te growth of ml·
`croorgan"i~ms by inhibiting their activity without n~\C£:S$nl'ily destroying
`them.
`.Meptio-lli,,fers to areas and practices where the int1>.nt is to i,,,
`st<l.rile.
`Aseptlc Processing-Those operations pcrfonnetl betw<>en th.(cid:173)
`st-erilization of an object or preparation and the finu! st>aling of ii.;; pack(cid:173)
`age. These OJ)<'rat-i,ms are. by definition, carried out in the compl.ete ab- ,j
`:1 ,
`sens~ of microorganisms.
`l
`Bactericide-Any agent that d.ast.roys microorganisms. _
`Bact-eriostat----Any agent th.at arrest.a or reta-rds the growth ofmi- '!,
`J'
`croorgani~m.s.
`Bioburden------T.h.c number of viable microorgani,;nu; present prior to i
`,!1--
`sterilization; Usually cxpre~e<l io colony-forming uni¼ of volume.
`Dii.infeetion- A process that decreases the probability of iofection !i ,_.
`hy destroying vegetative microorganisms. but not ordinarily bacteriaJ P ·
`sporns._ The t-erm usua-lly io applied to the u.:w of chemical agents on :.•. 1::t
`'! .
`inauimate-objects.
`Germicide'"--,An agent that destroys m:icroorgani~_ms, b\1t noL net-'i · ·
`e;<Sarily bacterial sp,)res.
`,
`,: , ,
` , ..
`Sanitization- A process th.at 1·edt1ces th<' level <>fbioburclen in oro.n __ i
`a product or object to a safe level.
`Sterile-The absolute absence of viable microorganisms. There.,: ,,,
`no degree <>1· partiality.
`·
`Sterility A&lur11nt,e Lovd (SALl-An est-im{1te 11f the dfecthe.l _
`ncse of a stcrili.zi>tion p:rocims. It usw11ly is Cl(pI'<Jssod in tenn~ <>f the: ''
`negative J)<)Wer of 10 (ie, l in 1 million = 10 ").
`! i (cid:173)
`i : ;:
`
`1.
`
`Sterilization is an essential concept in the preparation of ster(cid:173)
`ile pharmaceutical products. Its aim is to provide a product that
`is safe and eliu1inates the posi,ribility nf introducing infection.
`:Mergers and acquisitions of pharmaceutical companies cre(cid:173)
`ate multinational organizations faced with complying with all
`of the regulatory agencies of the involved countries. To date
`there is no global regulatory agency that oversees the produc(cid:173)
`tion of sterile pharmaceutical pruduets. Multinational eompa•
`nies must be familiar with the regulation,s of all countri.es .in
`which they operate and meet those regulations. Although it is
`not the intent of this chapter to delineate the sterilization stan(cid:173)
`dards for all countries, it is to provide a detailed description of
`the techniques used thrnughout the world to sterilize -pharma(cid:173)
`ceutical products. •rhere a.re many attempts to standardize
`practice8 throughout the miiltinational industry. These include
`the efforts of the Internati.onal Council on Harmonization (ICH)
`and thQ issuance of various tech11ical ISO standards and com(cid:173)
`pimdial efforts of the various countries like United States Phar(cid:173)
`n;uu:opo,iia.s (USP) t.o set s<>me basic standards. Additionally, or(cid:173)
`ganiiations like International Society for Pharmaceutical
`Engineering (ISPE) and Parente_ral Drug Association {PDA)
`have issued various documents. which include all facets of the
`inten1ational regulatory requirements.
`Sterilization is a process uMd to dt>,stroy or eliminate viable
`microorganh1rn~ that may be present in or on a particular prod_.
`uct or package. The process .requires an overall understanding
`and control of all parts of the p.reparation for use of a particular
`product. Thos.e areas include the selection and aL-eeptance of all
`materials used for the product and package, environment in
`which the product is prepared and used and the ultimate dis-(cid:173)
`position of the remaining materials after use. Sterilization may
`be required for several steps of the process using any one or a
`comhination of the techniques listed in this chapter.
`The aim of a sterilization process is to destroy o.r eliminate
`microorganisms that are present on or in an objeet or prepru:a(cid:173)
`tion, to make sure that this has been achieved with an ex(cid:173)
`tremely high level of probability and to ensure that the object
`or pi-eparation is free from infection hazard.~. The current-ly ac(cid:173)
`cepted pmfn.rmance target for a sterilization. process is that it,
`provide for a probability of finding a nonsterile unit of less than
`1 in 1 million. That is, the process (including production, stor-(cid:173)
`age, and shipment) will provide a Sterility Assurance Level
`(SAL) equal tonr better than 10•-s_ Tbis is achieved through the
`processing of product. in validated equipment and systems.
`Thorough validation and periodic requalification is essential to
`meeting these s terility requirement..-,,
`1'he purpose of this chapter is t(, provide a basic un.der(cid:173)
`standi.ng of the foll<>wing sterilization methods currently being
`
`776
`
`Novartis Exhibit 2024.003
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`.Sterililmtion-A proc;,s,; hy which all viable mi<:roorgimis,ns are
`removed or dBstroy~. baaed on a· probability function,
`Te:nninol Sterillzat,ipn- A process used to render products stei:ile
`to a preforrod SAL.
`Validation-The act of verii),-ing that a procedure is capable of pro(cid:173)
`duc&\g the inwnded result under pre~crihed circumstances and chal(cid:173)
`lenges to predefined sp€cificatio11s,
`Virieide-An agent that will desti·oy vi;ruses,
`
`STERILITY AS A TOT AL SYSTEM ·
`It is ne-cessary to reiterate the concept already brieflf"·ad·
`dressed in the introduction. The task of the technology we are,,
`dealing with is to provide the product in sterile conditions to the
`end us(l'r. It is currently ackriowl.edged !,hat the quality of the
`pmduct mu.st be built into the proce~s. This concept is particu(cid:173)
`larly true when one of the essential qualities of' the product is
`sterility,
`Accordingly, the above-mentioned task is accomplished with
`a series of design, ptoductinn, and dist.ribut.ion ateps that can be
`suimnarized as adivities for the selection and routine checking
`of the following items:
`• Active con&tituent-s, additives, raw m,aterial;; in general
`• Water used both as solvent and as wu&hlngirinsing agent
`• Pa~.kngin.i:: suitable fort.he product and for the sterilfantion pro(cid:173)
`cesij that will be used
`• Working en"ironxnent and equipment
`• Pen;onnel
`
`These pr()(:edures clearly have the purpose of providing the
`sterilii:ation process with a prod1.tct that has a minimum, defi(cid:173)
`nite, and consistent hioburden. There are also the following ac(cid:173)
`tivities:
`• Selection of the sterilization method that mQst stdt2 the unit
`formed by the product and its packaging, Slid definition oft.he pro(cid:173)
`cea, vuriubfos for obtaining the intended SAL
`• Selection of the machine that is most suitablo for performing the
`selected metliod and of the utilities that this machine requires
`• qualifi.eation and validation of tho machine and of' th.- pl'OC<Jss
`• Routine checldng of the process
`• Checking of the results of the sterilization proces;,
`• Propcr sforago of sterile goods und verification that their ste.riljty
`is maintained w,th full reliability throughout the allowed st-Ornge
`period
`• Delivering, opening, and using sterile goods without recont.ami•
`n.atfon.
`
`It also should be noted that, in December 2002, the US Food
`and Drug Administration (FDA) proposed new regulations for
`aseptic processing and terminal sterilization, The proposed
`rules as ddined in their Concept Paper require tlu1t. rnanufao(cid:173)
`turera of ;;t,erile p:rod\lcts use validar,ed and robust steriliiation
`techniques wherever possible. The European Pharmacopeia
`and related pharmacopeia_s have modified their requirements
`in their rulings identified as Annex L
`
`CONTAMINATION
`Certain fact,~ about microorganisms nmst be kept in mind when
`preparing sterile products-. Some microbes (bacteria, molds, etc)
`multiply in the refrigerator,. others at temperatures as high as
`60•C. Microbes vary in their oxygen r.equitemHnts from the
`st.net anaerobes that cannot tolerate oxygen to aerobes that de(cid:173)
`mand it. Slightly alkaline growth media will support the multi(cid:173)
`plication of many microorganisms while ot.hers flourish in
`acidic environments. Some microorganisms have t.he ability to
`use nitrogen and carbon dioxide from the air and thus can ac(cid:173)
`tually multiply in distilled water. In general. however, most
`pathogenic bacteria have, rather selective cultural require(cid:173)
`ments, with optimum temperatt1res of ,\0° to 37•0 and a pH of
`7,0. Cont,aminating yeasts and molds can develop readily in
`glocose and other sugar solutions.
`
`CHAPTER 40.: STERILIZATION
`
`777
`
`Actively growing microbes are, for the most part, vegetative
`forms with little resista,nce to heat and disinfectants. However.
`some foxms of bacteria-among th.Hm the bacteria t,hat cause
`anthrax, tetanus, and gas gangre.ne-have tbe ability to as-
`1mme a spore state that .is very.resistant to heat as well as to
`many disinfectants, For this reason, an excellent measure ()f
`succ.e'Ssful sterilization is. whether the, highly resistant. spore
`forms ofnonpatliogenic bacteria have been killed,
`'.rhenature rJfexpected contamination and the bioburden are
`important t-0 pharmacists preparing materials to be ate.rilized,
`The raw materials they work with rarely will be sterile, and im(cid:173)
`proper storage may jncrease the microbial con tent, Because the
`pharmacist seldom handles all raw materials in a sterile or
`prot.eeted 1mvitomuent, the enviromnental elements of the
`manufacturing area (air, surfaces, water, etc) can be e~pected
`to contribute to the contamination of a preparation, 'l'he con(cid:173)
`taine.r O'r packaging material may or may not be presterilize.d
`. and thus may contribute to the total microbial load.
`Understanding the nature of contaminants prior to sterd·
`ization and application of methods for minimizing s uch con(cid:173)
`tamination is vital to prepa1·ing for successful pharmaceutical
`sterilization, Examples of such methodi; include:
`• Ma111tena11c,; of a hygienic laboratory
`• Frequent disinfection of floors and surfaces
`• Minimization of traffic• in an.d oat of the area
`• Rofrigorated sto~age of raw materials and preparations that sup(cid:173)
`poJ·t microbial growth
`• Use of laminar aiJ·flow devices fot· certain criti,:aJ operations
`• Use of water that is of appropriate USP quality and i$ free Qfmi•·
`crobfal contamination (It is preferable to use presterilized water
`to avoid uny possible contamination.)
`
`METHODS
`General
`The procedure to be used fot stetilizing a drug, a pliarmacenti•
`ca.1 prepar~tion, or a medical device is determ.ined to a large ex.(cid:173)
`tent by the nature of the product. It is important to remember
`that the ~ame sterilization technique cannot be applied univer(cid:173)
`sally because the unique properties of some materials may
`result in their destruction or modi6cation, l\fothods of inacti(cid:173)
`vating microorgariisms may be da..,sified as either pbysfofJl {Jr
`chemical. Physical methods include moist heat, dry beat, and
`irradiation. Sterile filtration is another process, but it only re(cid:173)
`moves, not inactivates, microorganisms. Chemical methods in•
`elude the use of eithe1' gaseous or liquid sterilants. Guidelines
`for the use ofman,y types ofindustrial and hospital ste.rilization
`am available, 1- 10
`P;ach sterilization method can be evaluated using experi(cid:173)
`mentally derived values representing the general ina.ctivation
`rates of the procl'.SS. For exm11r,le, a dt~ath ·m te or survival curve
`for a standardized species can be d.ia,gramed for different ster(cid:173)
`ilization conditions. This is done by plotting the logarithm of
`surviving organisms against time of exposure to the steriliza(cid:173)
`tion method. In most instances, these data show a linear rela(cid:173)
`tionship, ty:pieal of first-order kinetics, and suggest that a
`,com,tallt proportiou ofa contaminant population is inact.ivated
`in, any given time intei:vaL .Based on such inactivation curves.
`it i1o possible to derive values that represent the general inacti(cid:173)
`vation rates of the process. For exan1ple, based on such data, it
`has become common to derive a decimal .reduction time or D
`value, which represents the time under a stated set of steriliza(cid:173)
`tion exposure conditions required to reduce a s urviving micro(cid:173)
`bial population by a factor of 90%.
`IJ val.ues, or other expressions of ste.rihzation process rates,
`provide a means of establishing dependable sterilization cycles.
`Obviously, the initial microbial load on a product to be steril(cid:173)
`ized becomes an important consideraiion, Beyond th.is, how(cid:173)
`ever, kinetic data al$o can be UI;ed to provide a su;r.tistica] basis
`for the su,ccP,ss of sterili.zat:ion cydes, A simple example wi.11
`
`Novartis Exhibit 2024.004
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`778
`
`PARr S: PHARMACEUTICAL MANUFACTURING
`
`0
`
`10"
`10'
`!O' .---.-~,.,.
`10'
`1 o' . im.>®"'ov .. .,.
`NegaOve-(No Growth)
`Thorrno··Chemical
`,
`1
`O · P'os;tive-(Growthl "'--.... Death Time
`••.--1~--.. 1-, .... , -
`, .. ....,. ...... ,. .... ,~
`10
`20
`30
`·.40
`., 10••., S . 15
`25
`35 ' ,,
`',,,
`.!!! !0''·7
`Exposure llme
`= .3 :
`lo Have a
`" ,
`·.g lO ·1
`' ,,
`10·• Probability
`"8 1Q'·i
`. . "-, of Survival
`a: 10, -; Probat5ility of One Organism Surv1\/1ng"•,, ,
`/
`,. ..... ,
`1 od ..... ,._ .... , ...... ,.....-,-...,..-,--.
`' '
`75
`55
`45
`5
`1 5
`25
`35
`65
`30
`40
`50
`60
`70
`Time-Mrnutes
`
`10
`
`20
`
`80
`
`tions of the chosen microorganism. Depending on the initial
`hypothesis of exposm·e at constant temperature, each D value
`always refore to a specified tert:1perat\ire,
`Equation l allows one to draw a first very irnportant condo,,
`sfon: the time requfred to reduce the microorganism concentra(cid:173)
`tion to any preset value is the function of the initial
`concentratio11. The sterilization ni.action is therefore neither an
`all-or-,wthing process nor a p<1tential barrier process as was
`once thought.
`It a]$O is evident immediatelv that the offed of st.erilization
`at the same constant temperature will be very different de(cid:173)
`pend.ing on the D value of t,he contaminating microbial species
`(or on the largest D value in thH u.;mal case of mixed c~mtami(cid:173)
`nation). Figure 40-2 shows that t hi; same reduction ratjo for dif(cid:173)
`fornnt .species is achieved after exposure time propo1·tional to
`the D value of each species. The graph derives only from Equa(cid:173)
`tion 1 and from the definition of D value. The basic hypothesis
`of fh(l t(impemture being constant, is thoroughly valid.
`Sterility Is a Probable Effect of Exposure Time-Let u& now
`consider what happens v.ithio a batch of units (vials , bottles,
`or others) with an in.itial constant unit contamination of lOQ
`m.icrnorganifnns equal to 102
`. If the D value at 12l "C i1> as(cid:173)
`sumed to be 1, after 1 min at 121°C, a reduction equal to 101
`,~ 10 microorganisms is achieved.; after another minute, only
`10° ,~ 1 microorganism is still surviving, After another
`minute, the surviving mfrrol;ial population would be 10" 1 ~
`1/10 microorganism. A contamination of 1/10 must not be tin·
`derstood to mean that each unit contains 1/10 of a microor·
`ganism, which is biologically meaningless (in this case the
`unit probably would be sterile) but that there is a probabiHty
`of having 1/10 of the units still contawinated within the batch
`of st<a>.rilized units.
`In fact, 3 min would be t he necessary time to redo<'.t' the
`:microbial population to a single surviving microorganism if
`the initial population were 10 times larger than the one at is(cid:173)
`sue. This higlwt· 'initial contamination could be regarded ei,
`ther as a 10 timefl larger numl>er of microorganisms in the
`same unit, or as the initial contamination of a 10 times large,r
`unit.
`If the unit L~ not considered any longer as the single vial ot
`bottle, bt1t as the whole of all the items produced O\•er a period
`of ti.me, t.lie initial number of mi~"roorganisms pres.ent in eath
`item has to be multiplied times the number ofi'tems produced, ,
`and the exposure time to achieve the reduction to the same
`number of viable microorganisms left in the whole of ihe items
`·produced, has to be increased correspondingly. The following
`(1X~ltrrple w.ill be helpfiil to focus the matter.
`A new sterile product in ampules has to be manufactured[
`·
`the number of ampules to be produced over all the life pe ·
`the product is expected to be 101.0. The ma:ximum nuh1
`contaminated amr>ules deemed to be acceptable is 10 "" 1:
`obviously means that the pwbability of having non.sterile
`
`pules aft.er sterilization must.not exceed 10-10, Let. us also
`·pose that the microbial population within each a,rupule afte
`filling and the sealing does not exceed 103 mkro
`These mu!'t be destroyed by means of moist heat-terminal
`ilization at 121 •c. The applicable D val.u1i is 1 min, The
`number of microorganisms to be desi royed during the life of
`product will be
`
`1010+3"' 1013
`
`If this wholt} nunobial population Wffft~ exposed to tnoist
`at 121 •c ove.r a period of ·13 min, it woukl be reduced to 1
`time.sits initial numbei- (ic, to 1013- rn = 10° = ,1. Thee .
`time of 13 min thus would be sufficient (under all the ·
`above hypothes es) to prevent the total number of contami
`ampules from exceeding the vr,Lue of 1.
`From the point of view of each single ampul.e, 13 min
`posure would reduce the microbial population to the theqre
`value of
`
`Figure 40-1. Sterihz.ation model u,ing D values.
`
`suffice (Fig 40-1). When the initial microbial contamination
`level is assumed to be 10°, and if the D value of the sterilization
`process is 7 m.inut-0s, r,omplet-0 kill is approached by apr,licatfon
`of6 D vah.es (42 minutes) . .However, at this point reliable ster·
`iliiation would not be assured because a few abnormally resis(cid:173)
`tant members of the population may remain. In this example,
`by extending the process to include an additional 6 D values,
`most of the remaining population is inactivated, reducing the
`fJrobability of one orgauism survivi1)g to one in l milli@,
`
`Moist Heat
`
`ESSENTIALS OF STEAM STERILIZATION KINETICS
`Let us 1;t1ppos1;, a system contaminated by m.ic.roorgan.isms
`(which we assume, for the sake of simplicity, to be pure and ho(cid:173)
`mogeneous) is immersed in pressurized saturated steam, at
`co11.~tant. temperature; for exam pk\ it could be a vial containing
`an aqtuiow; s 11spension of a certain spore-forming microor(cid:173)
`ganism.
`It has been shown expe,rimentally that, under the above con(cid:173)
`ditions. the reaction of thermal degradation of the microorgan(cid:173)
`ism obevs the laws of chemical reactions: the rat.e ofreduction
`of the n~mber of microorganisms present in the system in each
`m,,ment is proportional tn the uctnal nurnl;el' itself. The pro(cid:173)
`port.ionality coeUi.cient .is typical of the species and condit..ions of
`the chosen microorganism,
`Thus, the degradation rnactirm (the sterilization process) d(!·
`velops like a first-order dum1ic:\l reaction in which the reaction
`rate is proport.ional, in each moment, only to t he amount oi: mi(cid:173)
`crnorganisms still to be inat,tivated. This seems to be obvious
`for dry sterilization. but less rigorous for steam sterilizaticln, in
`which the water vapor molecules also seem to take part in the
`react.ion. Actually, this bimolecular reaction is of the fin1torder,
`as the steam is IJresent in high excess during the on ti.re reaction
`and its con.c.entralfon may be regarded as constant.
`, 'rhe most frequently used mathematical expression of the
`above facts is
`
`N = No 1.0"uv
`
`(l)
`
`where N 0 is the initial number of microorgat)isms, t is the
`elapsed exposu re. (equal to ste.rilization iime), N is the n.uml~r
`ofi:nicr(lorganisms after the exposure time t, and Dis the deci(cid:173)
`mal deca.y time; defined as the time interval required, at a spec(cid:173)
`ifled constant temperature, to reduce the microbial population
`being considered by 1/10 (ie, by one logarithmic. value; eg, from
`100% to 10% or from 10% to 1% of the initial vafoe),
`The D value is inversely-proporticmal to the first-order rear(cid:173)
`tion ~oeflkien t and is therefore typical oft.he specie& and con di-
`
`Novartis Exhibit 2024.005
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`CHAPTER 40: STERILIZATION
`
`179
`
`interpret (~s numeric value as the probability of st:iU hav(cid:173)
`g one contaminated. ampule in 10 billion sterilized ampules
`eans that a single ampule will still be contaminated out of a
`le lot of 1010_ This probability value is defined as PNSU
`robability of nonsterile unit),
`In recent times the PNSU as a sterility evaluation criterion
`being replaced by the SAL. The name itself could generate
`me misunderstanding, because a level of assurance com(cid:173)
`ly is deemed to be good if high, but SAL seems to have been
`ned in such a way that its numerical value is the same as
`SU. This noffiithstanding, it is sometimes calculated as the
`iprocal value of PNSU. The SAP (sterility assurance .pmba(cid:173)
`lity) criterion has been proposed as well and SAP seems for.
`themomeu,t to have been granted the same definition of PNSU,
`'even if it would be better understandable if its value ap(cid:173)
`proached unity after a satisfactory sterilization.
`The above discussion and example lead to the conclusion that
`the optimum exp0sure time for a sterilization process must take
`into account not only the initial microbial population within the
`single item to be sterilized and the species and conditions of the
`contaminating microorganism, but also the total number of
`items expected to be sterilized over the lifo of tho product.
`.
`Effect of Temperature Changes-All the above. considera•
`Ii: tfons have been developed under the basic assumption that the
`'" temperature is kept constant during the entire exposure time.
`I It seems rather obvious that the D value will change as the tem-
`perature changes, If the D values experimentally obtained for a
`given microbial species are plotted on a semilogarithmic chart
`. as th-0 function of the temperature T, a path similar to I<'igure
`40-.'l is obtained.
`In this case, it can be seen that D value is 1 min at 121 •c (i-0,
`the average value which very often is assumed to be acceptable
`in the absence of more exact. e~perimental data). It also can be
`
`seen that D value varies by a factor of 10 if foe temperature
`varies by 10•c.
`The z value fa defined as t.he temperat ure coefficient of mi·
`crobi.al d-0struction, the number of deb,rees of temperature that
`causes a IO-fold variation ofD for, more generally, of the steril(cid:173)
`ization rate). The i values generally oscillate between 6 and 13
`for st.earn sterilization in t he range 100• to rno•c, and z value
`often is assumed to be equal to 10 in the ahstmce of more pre•
`dse experimental d.ata.
`The fact that D value varies by 10 times for a variation of
`1o•c wh-0n z ~ 10 mu.<it not lead to the false assumption t:hat D
`varies by one time (ie, doubles) for an increase of 1 •c. Obv.iously,
`thi.~ is not true, It is actually a nrntter of finding tbe mimber·
`which yields 10 when raised to the tenth power, This number is
`.1.24 .. Therefore, a variation of 1 •c entails a variation ofD value
`of24%. This is quite a significant number, which illustratell the
`dramatic effects that are. generat~.d when the sterilization ten1·
`perature is also ()nJy a few degrees· lower than the expected
`value, perhaps only in some areas of the i,i;erilizer load.
`It is also useful to reme,mber tha t the effect of temperature
`variation decreases considerably as t he temperature ri$es and
`drops to approximately 1/2 (or eVf:U less) for dry sterilization at
`approx:imately 2oo•c. 'Under these conditions the z value is
`about 20 instead of about 10. Therefore, t.he small temperature
`differeuces that can be so dramatic in steam sterilization ha ve
`much less effect in dry sterilization,
`The foregoing refers to ave.rage values because the actual D
`values and z values depend to a large extent on the medium
`t,hat contains t h.e microorganisms and on their history. At
`121 •c no mkroorganis m has exactly D "" 1 and z "' 10, How•
`ever, the combined use of these two parameters in calculating
`F0 and PNSU provides ample margins of safety with regard to
`the m.icroorgauisms with which we deal commonly.
`
`Number of microorganisms per unit
`1CJA2
`
`1QA(-1)
`
`10"(·2)
`
`1(}A(-3)
`
`1(}A(-4)
`
`1 (}A(-5)
`
`... "'lit
`16
`
`10"{-6) 0
`
`1
`
`5
`4
`3
`2
`Steri~zation time (minutes)
`
`6
`
`7
`
`8
`
`figure 40-2. Effect of varying D values on steriliiation rate (courtesy, f edegari Autod avi).
`
`Novartis Exhibit 2024.006
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`780
`
`PARTS: PHARMACEUTICAL MANUFACTURING
`
`Number of microorganism per unit
`10•2
`
`z=1o·c
`1 QA(-1) .___....__ _ __._ _ ____. __ ......_ _ _._ __
`121
`126
`116
`131
`100
`111
`101
`Sterilization temperature (·C)
`
`Figure 40-3. Effect of temperature on microbii!I destruction {courtesy, Fedegari Autociav1).
`
`Foor Equfoalent Sterilization 7'ime a.t 121°C-1t is of th!! ut(cid:173)
`most interest to calculate thf;! lethal effect of the exposure of 1.1
`mi<..·r obial population fo a variable tempc,n,ture, T, by relating it
`to an hypothetical sterilization performed at a constant temper(cid:173)
`atuni, To, for the time, to. If the co11stant reference teiµperature
`is assumed equal to 121.l°C (originally 2/.i0°F') an d the z value
`equal to 10, the equivalent, time is termed 1"0 , Thus, F'o .is the
`equivalent exposm-e time at 121.l°C of the actual exposure time
`at a variable t emperature, calculated for .µ1 ideal microorgan(cid:173)
`ism with a temperati1rn coefficient of destruction equal to 10.
`First introduced in the Laboratory Manua./. for Food Canners
`and Processors by the National Canners Association in 1968, F 0
`has become a common term .in pharmaceutical ptod uL'tion since
`tbe FDA used it extensively in the "Proposed Rules" of June 1,
`1976 (21 CFR 212.3) with the following meaning:
`Fo meana the equiv-ulent amount of time, ln minult-s at 1.21,l"C
`(250'.F). which has been delive:red to a product by the sterilization pro(cid:173)
`cess .. For the calculation ofit, a z value of 10°C 01· 1e•.F is aesumed; the
`term z value means the slope of the therm.al death t-in,e cnrve and may
`be expressed as the number of degrees required to bring about a 10-fold
`change in the dealh rut:e.
`l n practice, the knowfodge of the tellrpernture values as the contin(cid:173)
`uous function of elapsing time is 11ot availa.hle, and F0 is calculated M
`1'-·· 121.l
`-
`F0 '" At. l 10 -
`z
`where l!.t is the time interval between two consecutive mea(cid:173)
`surements of T, Tis the temperature of the st-erilized product
`at time t, and z- is the temperature coefficient, assumed to be
`equal to 10.
`
`(2)
`
`low pliiut and running costs. The sterilizing medium obviou1;ly
`is pressurized saturated steam and the typical operating tem(cid:173)
`perature is 12.1 •c (250°F), but. h igher or lower temperatures of(cid:173)
`ten are used.
`The term dry saturated steam sometimes is u~ed; it should
`be nJade dear t hat th.is is an ideal condition of st.Jato, and that
`moist t:!aturated. steao:i is ustid in practice for sterilir,ation. How(cid:173)
`ever, the steam must entrain the smallest possible amount of
`condensate. The water oapor ratio of the steam defines the
`amount of condensate entrained by 100 parts by weight of moist
`steam; a water vapor ratio of 0.95 means that 100 g of steam
`cons.ist of 95 g of dry saturated steani plus 5 g of condensate
`which is, or should be, at the same temperature as the steam.
`The reliability of sterilization performed with saturated
`steam is based Ott several particular characteristics of this
`medium, \X,7_1e.n steam condenses, it releases calories at a con•
`!Stant temperatl)r.- and in a considerable amount: 1 kg of pure
`saturated steam condensing at 121°c (turning into water at
`121 •c, thus without cooling) releases as much as 525 kcal. The
`t-Omperaturos and pressures of saturated steam have a two-way
`correlation. On<~e the tetripl.".rature of t,he steam ill determiued.,
`1:10 is its presi:mre, and vice uersa. S11tural:ed st!;lam at 121°C in(cid:173)
`evitably has a pressure of2.05 abs bar, This entails two very in.·
`teresti:ng, practical possibilities:
`l. A pure !i>aturnted ~team uutodave cau oo con'frolled indifferently
`according t-0 the 1emperat.ure parameter or according to the p.res•
`s.ure p(l..raineter,
`2. 'R,egardle&~ of th.e parameter used for eontrnl, the second parame0
`ter c;m be. used easily to cTOs$-monitQr the first one.
`
`Saturated Steam
`
`PRINCIPLES
`Sterilization with saturated. steam is the m ethod that provide.s
`the best combination of flexibility in operation, safe r esults and
`
`A 1 gram molecule of water (18 g, or 18 mL in the liquid state)
`as steam at 121•c and 2.05 abs bar occupies a volu