as) United States
`a2) Patent Application Publication co) Pub. No.: US 2012/0091026 Al
`(43) Pub. Date: Apr. 19, 2012
`
`Chacornacetal.
`
`US 20120091026A1
`
`(54)
`
`METHOD OFSTORING A VACCINE
`CONTAINING AN ALUMINUM ADJUVANT
`
`(75)
`
`Inventors:
`
` a
`Isabelle Chacornac, Tupin E
`Semons (FR); Nabila
`Ikhelef-Gribi, Francheville (FR);
`Frédéric Ronzon, Montromant
`(FR); Julien Tirefort, Lyon (FR):
`Sandrine Lentsch Graf, Sainte Foy
`Les Lyon (FR)
`
`(73)
`
`Assignee:
`
`SANOFI PASTEUR,Lyon cedex
`(FR)
`
`(21)
`
`Appl. No.:
`
`13/274,714
`
`(22)
`
`Filed:
`
`Oct. 17, 2011
`
`Related U.S. Application Data
`
`(60)
`
`Provisional application No. 61/454,248, filed on Mar.
`18, 2011.
`
`(0)
`
`Foreign Application Priority Data
`
`CFR) occ teeeeeesrentene 1058464
`Oct. 18, 2010
`Publication Classification
`
`(51)
`
`(52)
`67)
`
`Int. Cl.
`(2006.01)
`B6SD 85/00
`(2006.01)
`BO6SB 7/28
`VS. C1. ceccccccccet cee certeseeeeeseeceee 206/524,3; 53/471
`ABSTRACT
`
`The invention relates to a method for loading and storing a
`vaccine composition, containing the antigen adsorbed on the
`aluminumadjuvant which (a) comprises (i) loading the com-
`position into a container; and(ii) closing the container witha
`device in particular acting as a stopper, the surface of the
`device getting into contact with the composition being coated
`with a fluoropolymersuch as Teflon™;and/or(b) loading the
`composition into a container wherein the inner surface of
`whichis coated with polymerized silicone. The use of fluo-
`ropolymeror polymerized silicone optimizes the adsorbed
`antigen stability upon storage. In a particular embodiment,
`the antigen is the hepatitis B surface antigen and the alumi-
`num adjuvant is aluminum oxy hydroxide.
`
`Regeneron Exhibit 1014.001
`
`

`

`US 2012/0091026 Al
`
`Apr. 19, 2012
`
`METHOD OF STORING A VACCINE
`CONTAINING AN ALUMINUM ADJUVANT
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`
`[0009] Whatever the container used for storage and the
`device used for closing the container, administration at the
`time of injection consists of using a syringe and sliding the
`plungeror the stopper/plunger combination so that the vac-
`cine is delivered.
`
`SUMMARYOF THE INVENTION
`
`‘Lhis application claims the benefit of U.S. provi-
`[0001]
`sional application 61/454,248,
`filed Mar. 18, 2011, and
`French patent application no. 10 58464, filed Oct. 18, 2011.
`The entire contents ofboth of these applications are incorpo-
`rated by reference.
`
`BACKGROUNDOF THE INVENTION
`
`1. Tield of the Invention
`[0002]
`[0003] The present invention relates to a method for reduc-
`ing and/or decelerating the desorption of an antigen that has
`been adsorbed on an aluminum adjuvant as well as to the
`product thereof—namelythe combination of a vaccine com-
`position comprising the antigen adsorbed on the aluminum
`adjuvant and a container for the composition, said container
`having particular characteristics.
`[0004]
`2.Summary of the Related Art
`[0005] A large numberofantigensare able to adsorb on an
`aluminumadjuvant, in particular at neutral pH or at a pH close
`to neutrality, which is the pH naturally required for compo-
`sitions that have to be administered to mammals, including
`humans.
`
`[0010] We have now foundthatthe material ofthe container
`itselfas well as that ofthe device for closing the container can
`affect the adsorptionrate.
`[0011] We observed that a vaccine composition containing
`the hepatitis B surface antigen (HBsAg) adjuvanted with an
`aluminum adjuvant and loaded as a single dose in standard
`syringe closed with a standard stopper underwent different
`adsorption rates when stored under identical conditions and
`for the same period of time (a few days to several months)
`depending on whether the syringe wasstoredin the vertical or
`horizontal position. The same phenomenon has also been
`seen with vials. The practical consequence of the different
`storage positions was that the vaccine contained in the reser-
`voirs stored vertically was not in contact with the stopper,
`whereas there was contact between the composition and the
`stopperin the horizontally stored reservoirs.
`[0012] Aftera certain period oftime,the level ofadsorption
`of HBsAg was measured, and we observedthat the level of
`adsorption was much lowerfor the HBsAg contained in the
`horizontally-stored reservoirs. ‘his indicated to us that the
`material of the standard stopper (chlorobutyl or bromobutyl
`polymer) was responsible for the adsorption decrease.
`[0013] The solution to this problem is either to prevent
`contact between the adjuvanted vaccine composition and a
`stopper made of a material that contributes to decreased
`adsorptionor to use a stopper made of a material that does not
`contribute to decreased adsorption. In one embodiment, the
`invention comprises a method and apparatus using a stopper
`coated with a film of a fluoropolymer, such as Teflon™or a
`Teflon™—\like substance (such as Omniflex™ from Helvoet
`Pharmaor[‘luorotec™ from West Pharma), which decrease
`or eliminate desorption caused bythe stopper.
`
`DETAILED DESCRIPTION OF THE INVENTION
`
`Inoncaspect, the invention comprisesa first method
`[0014]
`for reducing and/or slowing down the desorption of ananti-
`gen initially adsorbed on an aluminum adjuvant during stor-
`age, the method comprising (i) loading a container with a
`vaccine composition containing the antigeninitially adsorbed
`on the aluminum adjuvant; and (ii) closing the container with
`a device acting as a stopper, the surface of the device con-
`tactable with the composition being coated with a fluoropoly-
`mer.
`
`Provided that the amount of adjuvant is suchthat the
`[0006]
`antigen can actually adsorb on the adjuvant in an optimum
`amount when the two compounds are mixed together, the
`maximumdegree of adsorption is very frequently achieved.
`However, over time, depending on the environmental condi-
`tions, the percentage of adsorbed antigen (adsorption rate)
`may decrease, and this desorption mayconstitute an instabil-
`ity factor.
`[0007] Known environmental conditions that can affect the
`percentage of adsorbed antigen (adsorption rate) include, for
`example, variations in pH (evenslight variations), and the
`addition ofone or more medium component(s) or one or more
`additional antigen(s) that may compete with thefirst antigen
`for the adsorptionsites on the adjuvant.
`[0008] Conventionally, a ready-to-use multi-dose vaccine
`compositionis loaded intovials, e.g., glass vials closed with
`a plastic stopper. Similarly, a single dose of a vaccine com-
`position may be loaded in a mono-dosevial or a ready-for-
`injection syringe consisting, in a standard manner,of a reser-
`voir containing the vaccine, a plunger that closes the reservoir
`at ils distal end, and a device for administration, such as a
`needle attached at its proximal end. According to an alterna-
`tive standard filling mode, the vaccine dose may also be
`loaded in a needleless syringe to which the practitioner adds
`Inother words, the inventionrelates to a first method
`[0015]
`a separately packaged needle at the time of the injection. ‘he
`for fillmg and storing a composition containing the antigen
`reservoir of the syringe is generally madeofeither glass or
`adsorbed on the aluminum adjuvant which comprises(i) fill-
`plastic and the plunger or the stopper is simply made of
`ing a container with the vaccine composition; and (ii) closing
`plastic, such as a chlorobuty] or bromobuty! polymer, without
`the container with a device acting as a stopper, the surface of
`particular lamination. Standard glass or plastic syringes are
`the device contactable with the composition being coated
`sold, for example, by Becton-Dickinson; Gerreshcimer A G,
`with a fluoropolymer.
`Schott A G, Germany; Nuova Ompisrl, Italy; and West
`[0016]
`Ina similar manner, the inventionalso relates to the
`Pharma/Daykio. In order to facilitate sliding, the plunger or
`use of a device acting as a stopper for closing a container
`stopper mayhave been immersed in a silicone-in-water emul-
`sion so that a silicone film is formedat its surface. Standard
`containing a vaccine composition comprising the antigen
`adsorbed on an aluminum adjuvant, the surface ofthe device
`plungers/stoppers are sold by Helvoet, Stelmi and West
`Pharma, for example; some ofthem alreadybeing sold coated
`contactable with the composition being coated with a fluo-
`
`withasiliconefilm (ref. B2 from West Pharma). ropolymer.
`
`Regeneron Exhibit 1014.002
`
`

`

`US 2012/0091026 Al
`
`Apr. 19, 2012
`
`[0017] The container may be, for example, a vial or the
`reservoir ofa syringe.Thisalso appliesto all the other aspects
`of the invention described hereinafter.
`
`container, such as by sliding the plunger of a syringe, it is
`recommended to siliconize the innersurface of the container.
`
`
`
`
`[0024] However, it has been observed that silicone may in
`‘lhe vaccine composition maybe liquid orsolid,e.g.
`[0018]
`somecases be detrimental to adsorption. Indeed, the desorp-
`lyophilized. A lyophilized composition mayhave the appear-
`tion rate observed in compositions stored in syringes conven-
`ance of a powder. At the time of injection to a patient, the
`tionally siliconized by mere surface-treatment withasili-
`lyophilized composition is reconstituted. with an appropriate
`cone-in-water emulsion may be higher than the desorption
`pharmaceutical solution. This also applies to all the other
`rate observed in compositionsstored in non-siliconized con-
`aspects of the invention described hereinafter.
`tainers. We postulate that althoughthesilicone adheresto the
`[0019] According to conventionalpractice in the art and for
`inner surface of the container, it remains in free form and,
`he purposesherein, it is understood that “antigen adsorbed”
`uponshakingorstirring, can flow away fromthe inner surface
`or “initially adsorbed”is not intended to meanthat 100% of
`and pass into the container’s content (the vaccine composi-
`he antigen amountis actually adsorbed.‘hese terms simply
`tion).
`mean that a substantial amount ofantigen is adsorbed. This
`[0025] We have now foundthat this latter problem can be
`also appliesto all the other aspects ofthe invention described
`hereinafter.
`solved by using a container wherein the inner surfaces are
`coated with polymerized silicone. Such a container can be
`[0020] As mentioned above, the device may bea plastic
`obtained bytreating the inner surface of the container with a
`device madeoutof, for example, a chlorobuty] or bromobuty!
`silicone-in-water emulsion, followed by heating the con-
`polymer. This standard device is treated with a fluoropoly-
`mer; in particular, it may be submitted to a laminar flow
`tainer, for example at a temperature of 270 to 330° C. for 30
`reatment with a fluoropolymer, this laminar flow treatment
`min. Upon heating, the silicone polymerizes on the inner
`being carried out on the entire device or, at the very least, on
`surface of the container and is therefore no longer capable of
`he surface ofthe device contactable with the composition.
`mixing with the composition. Polymerizing the silicone
`The laminarflow treatment makesit possible to deposit a very
`makesit possible to reduce the surface energy ofthe silicone
`hin layer(e.g., film) of the Muoropolymer. As will be appre-
`to which the vaccine composition may besensitive.
`ciated, the area of the coated surface may exceed the surface
`[0026] Additionally,the siliconizing operation comprising
`contactable with the composition. Indeed, in one embodi-
`a polymerization step (1) is more precise and more homoge-
`ment, the whole surface of the device is coated with the
`neous that a simple standard siliconizing operation; and(ii)
`flucropolymer.
`makesit possible to reduce the amountofsilicone that is used
`[0021]
`For use in the present invention, the fluoropolymer
`(that is, loaded on the inner surface of the container) by about
`may be, for example, polytetrafluoroethylene (PTFE), poly-
`a factor of 10 without any loss of lubricating effect. For
`ctrafluoropropylene (PTFP), fluorinated ethylene propylene
`example, according to a standard siliconizing process, from
`TEP, a copolymer ofhexafluoropropyleneandtetratluorocet-
`400 to 1000 pg ofsilicone are deposited in a syringe intended
`hylene), polychloratrifluoroethylene (PCTFE), perfluaro-
`to contain doses of 0.5-1 ml (the total inner surface of the
`alkoxy co-polymer (PFA), poly(ethylene-co-tetrafluoroeth-
`0.5-1 mlsyringe reservoir is about 8 cm”; in this examplethis
`ylene)
`(ETFE),
` poly(ethylenechlorotrifluoroethylene)
`
`surface correspondsto an amountofsilicone offrom about 50
`ECTFE), polyvinyl fluoride (PVF) or polyvinylidene fluo-
`to 125 ug/em*), whereas from 40 to 100 ug of silicone are
`ride (PVPF).
`sufficient for the same syringe (about 5 to 12 wg/em?) if
`[0022] The method/use disclosed herein makesit possible
`‘o reduce the desorption speed ofthe antigen adsorbed on the
`silicone is deposited on the innersurfaces ofthe container and
`aluminum adjuvant and/or the desorption percentage (or des-
`then polymerized, for example by heating. The fact that the
`orption ratc) after a defined storage time at a given tempera-
`inner surface of the syringe is coated with a low amount of
`ture. The desorption rate may be expressed as follows:
`polymerized silicone in a more homogenous manner than
`with a low amount of free silicone allows non-siliconized
`amount of non-adsorbed antigen)/(total antigen amount
`present in the composition). Typically, the desorption rate can
`plungersto slide smoothly, whereas such plungers are inop-
`be assessed by centrifuging the vaccine composition
`erative with syringes coated with low amountoffree silicone.
`samples at T (time)—0 and at the end of the experiment);
`[0027] This is the reason whythe invention also comprises
`recovering the supernatants which contain the desorbedanti-
`a second method for reducing and/or slowing downthe des-
`gen; and then quantifying the desorbed fraction by assaying
`orption ofan antigen adsorbed on an aluminum adjuvant, the
`the antigen in the supernatantsand in the whole vaccine using
`method comprisingfilling a container with a vaccine compo-
`a suitable method chosen according to the nature oftheanti-
`sition comprising the antigen adsorbed on the aluminum
`gen. The desorption percentage (or desorption rate) can vary
`adjuvant, whercin the innersurface of the containeris coated
`from one antigen to another accordingto the strength/weak-
`with polymerizedsilicone.
`ness ofthe antigen-adjuvant interaction. Nevertheless, it is
`[0028]
`In other words, the invention also comprises a sec-
`consideredthat the desorption percentage(or desorptionrate)
`ond method for loading and storing a vaccine composition
`can be reduced by 10 to 15 or 20% compared with a standard
`containing an antigen adsorbed on the aluminumadjuvant,
`loading method using standard stoppers—said reduction
`the method comprising filling a container with the composi-
`being measured 1 or 2 months after the date of loading.
`tion wherein the inner surface of the container is coated with
`During this period of time, the storage is carried out at a
`polymerized silicone.
`temperature of+5 to 25° C. As maybeeasily appreciated, the
`adsorption percentage (adsorption rate) may be easily
`[0029]
`Inasimilar manner, the invention also comprises the
`deduced from the desorption percentage (or desorptionrate).
`use of a container having an inner surface coated with poly-
`[0023] Whenthe device is used not only to close the con-
`merizedsilicone for storing a vaccine composition compris-
`tainer but also to deliver the composition contained in the
`ing an antigen adsorbed on an aluminumadjuvant.
`
`Regeneron Exhibit 1014.003
`
`

`

`US 2012/0091026 Al
`
`Apr. 19, 2012
`
`after storage of the vaccine-containing containers at 25°
`C. for 2 months, starting from the date of filling the
`containers.
`
`the container having inner sur-
`[0030] Advantageously,
`faces coated with polymerized silicone is madeofplastic or
`glass. Advantageously, the container is the reservoir of a
`syringe.
`For the purposes of the present invention, the con-
`[0044]
`tainer may be any type ofreservoir, such asvials or syringes,
`[0031] The amount of polymerized silicone coated on the
`innersurface ofthe containeris from 3 to 25 ug/em?; advan-
`and may contain multiple doses (multidose container) or a
`single dose (single-dose container). As an example, the con-
`tageously from 5 to 20 ug/cm?; preferably from 5 to 15
`ug/em?.
`tainer maybea syringe or a part ofa syringe comprising the
`reservoir containing, the vaccine closed by a device acting, as
`[0032] As maybe casily understood, the container used in
`a stopper and asasystem for releasing the vaccineat the time
`the first methods of the invention may be advantageously
`ofadministration (e.g., using a plunger). The device acting as
`coated with polymerized silicone as described above.
`a stopper may be a plunger.
`[0033]
`In another aspect, the invention comprises:
`[0045]
`Stoppers and/or plungers for use in the present
`[0034]
`A—Acontainer (i) which contains a vaccine com-
`invention are sold, for example, by Helvoet Pharma (Omni-
`position comprising an antigen adsorbed on an aluminum
`fiex™ technology) and by West Pharma (Fluorotec™ tech-
`adjuvant; and (ii) which is closed by a device acting as a
`nology). Glass syringe reservoirs coated with polymerized
`stopper, wherein the surface of the device contactable with
`silicone for use in the present invention are sold, for example,
`the composition is coated with a fluoropolymer;
`by Nuova Ompisrl, Becton-Dickinson and Gerresheimer
`[0035] B—A container (i) having inner surfaces coated
`(Baked-on technology).
`with polymerizedsilicone; and (ii) containing a vaccine com-
`[0046] The antigen andthe aluminumadjuvant may be any
`position comprising an antigen adsorbed on an aluminum
`antigen and any aluminum adjuvant provided, of course, that
`adjuvant; and
`they are both capable of interacting with each other. Further-
`[0036] C—A container (i) having inner surfaces coated
`more, it may casily be understoodthat the present invention is
`with polymerized silicone; (ii) containing a vaccine compo-
`of particularinterest for an antigen-aluminum adjuvant pair
`sition comprising an antigen adsorbed on an aluminum adju-
`having, a relatively weak interaction force; the interaction
`vant; and(iii) whichis closed bya device acting asa stopper,
`force possibly depending on the environment. This interac-
`wherein the surface of the device contactable with the com-
`tion force can be assessed accordingto a varietyof tests. For
`position is coated with a fluoropolymer.
`example, an aluminum adjuvant may be used to form various
`[0037]
`In other words, the invention relales to a vaccine
`antigen-adjuvantpairs (the antigen varies from onepair to the
`composition comprising an antigen adsorbed on an aluminum
`other, the adjuvant remaining the same). Then a large amount
`adjuvant which is loaded into and stored in a container (i)
`of a compound able to compete with the antigen for the
`having the inner surface coated with polymerized silicone;
`interaction with the adjuvant is added. The various prepara-
`and/or (ii) which is closed by a device acting as a stopper,
`tions are centrifuged and the supernatants recovered. Finally,
`whereinat least the surface afthe device contactable withthe
`the amountofantigen desorbedis assayed in the supernatants,
`composition is coated with a fluoropolymer.
`and as a result, antigens may be compared for their interaction
`[0038] Vaccine compositions stored in containers accord-
`force with the adjuvant.
`ing to the invention include:
`[0047] A relatively weak interaction force is an interaction
`[0039] A vaccine composition comprising an antigen,
`force that leads to an adsorption that may be detrimentally
`wherein the minimal antigen amount required for
`affected bya standard filling with the composition containing
`intended use (e.g., as a dose for administration to a
`the antigen-aluminum adjuvant complex. Various elements
`involved in the manufacture of a container, such as, for
`human) is adsorbed on an aluminum adjuvant;
`example, latex, antioxidants, silicone and metal ions (e.g.,
`[0040] A vaccine composition comprising an antigen
`zinc and tungsten), can destabilize the antigen-adjuvant com-
`adsorbed on an aluminum adjuvant, wherein the vaccine
`plex.
`composition when loaded in a container for use in the
`present invention, exhibits an adsorption percentage of
`[0048] By “adsorption” it is generally meant any phenom-
`at least:
`enonaimed at forming, an antigen-adjuvant complex involv-
`ing i.a. electrostatic interaction forces, hydrophobic interac-
`tionsorligand exchange. Thus, the antigen may be attached at
`the surface of the network of the aluminum adjuvant or
`embedded inside after co-precipitation with the aluminum
`adjuvant.
`For usc in the present invention, an aluminum adju-
`[0049]
`vant maybe aluminum oxy hydroxide (AIOOI)), such as the
`product sold by Brenntag AG (Superfos) or Reheis Corp.; and
`aluminum hydroxyphosphate (AIOHPO,), such as the prod-
`uct sold by Alphos.
`[0050]
`For a vaccine composition to be effective, the mini-
`mal antigen amount required for adsorption onto the alumi-
`num adjuvant essentially depends upon the antigen itself, and
`is readily determinable by those ofordinary skill in the art.
`[0051]
`Ina particular embodiment, the antigen can be the
`hepatitis B surface antigen (HBsAg).It is particularly advan-
`tageous to adsorb HBsAg onto AIOOH as HBsAgexhibits an
`
`(a) 65-70% ofthe total antigen amount present
`[0041]
`in the composition, when, immediatelyafter loading,
`the compositionis stored al (1) 523°C. for 2-3 years;
`or (ii) 253° C. for 2-3 months; or
`[0042]
`(b) 80-90%,of the total antigen amountpresent
`in the composition, when, immediately after loading,
`the composition is stored at 543° C. for 18 months;
`and
`
`[0043] A vaccine composition comprising an aluminum
`adjuvant and an antigen able to adsorb onto the alumi-
`num adjuvant, wherein the percentage of the antigen
`adsorbed on the aluminum adjuvantts at least 5 or 10%
`higher than the percentage observed when the same
`vaccine composition is contained ina standard container
`having uncoated inner surfaces or inner surfaces coated
`with non-polymerizcdsilicone, the comparison between
`the antigen adsorption percentages being carried out
`
`Regeneron Exhibit 1014.004
`
`

`

`US 2012/0091026 Al
`
`Apr. 19, 2012
`
`iso electric point (IEP) less than 7 (about 4 to 5) and AIOOH
`exhibils a point of zero charge (PZC) greater than 7 (about 9
`to 11).
`[0052] The vaccine composition for use in the present
`invention can contain one or more antigen(s), at least one of
`them being adsorbed on the aluminum adjuvant andit being
`possible for the others to be adsorbed as well or not.
`[0053] According to one embodiment, the composition for
`use in the present invention comprises HBsAg adsorbed on
`AIOOH (AIOOH-HbsAg complex) and a second antigen,
`whichis polyribosylribitol phosphate (PRP) ofHaemophilus
`influenzae (HiB valence), preferably in a form conjugated to
`a carrier protein (C) which maybei.a. Dt or Tt.
`[0054] According to another particular embodiment, when
`the antigen adsorbed on the aluminum adjuvant is HBsAg,the
`vaccine composition mayalso contain, as additionalantigens,
`one or more ofthe following: diphtheria toxoid (Dt) (diph-
`theria valence); tetanus toxoid (Tt) (tetanus valence); Borde-
`tella pertussis detoxified toxin (Ptdx), fimbriae, filamentous
`haemagglutinin (FHA) and/or pertactin (69 kD antigen) (per-
`tussis valence); inactivated poliovirus serotype 1, 2, or 3
`(polio valence); and polyribosylribitol phosphate (PRP) of
`Haemophilus influenzae (IMB valence), preferably in a form
`conjugated to a carrier protein (C) which maybe 1.a. Dt or Tt.
`[0055] As a matter of example, the composition may com-
`prise HBsAg, Dt, Tt, Pt and FHA adsorbed on AIOOH (the
`AIOOH-HbsAg-Dt-Tt-Pt-FHAcomplex), the polio valence,
`and PRP-C substantially non-adsorbed on AIOOH.
`[0056]
`In a general manner, the inventionis also particu-
`larly advantageous when the vaccine composition comprises
`several valences, for example 2, 3, 4, 5, 6 or more, each
`represented by one or more antigens (2, 3, 4 or 5), several
`antigens being adsorbed on the aluminum adjuvant. Indeed,
`under the standard filling mode, the higher the number of
`antigens/valences adsorbed on the aluminumadjuvant, the
`more crilical is the phenomenon of destabilization by the
`container. The antigen-adjuvant interaction force often dif-
`fers from one antigen to another and, in a composition con-
`taining several antigens, the antigen with the weakest inter-
`action torce exhibits the highest tendency to desorb under
`adverse conditions.
`
`Examples and Experimental Results
`
`A—Abulk ofa vaccine composition containing the
`[0057]
`hepatitis B surface antigen, diphtheria toxoid, tetanus toxoid,
`and pertussis valence, each adsorbed on aluminum oxy
`hydroxyde, as well as the polio and non-adsorbed Haemophi-
`lus influenzae B (HiB) valences was distributed into three
`categories of single-dose syringes,
`the characteristics of
`which were as follows:
`[0058]
`(1) standard siliconized glass syringes with stan-
`dard stopper/plunger made of non-laminated plastic;
`[0059]
`(2) standard siliconized glass syringes with fluo-
`ropolymer-coated stopper/plunger (West Pharma; I'Ino-
`rotech™technology); and
`[0060]
`(G3) syringes, the inner surface of which is coated
`with polymerized silicone (Baked-on syringe system
`Luercone™ from Gerreshcimer) with fluoropolymer-
`coated stopper/plunger (West Pharma;
`I"luorotech™
`technology).
`[0061] The bulk was distributed in 0.5 mL single doses,
`each dose containing 10 ug of HBsAg, 30 Lf of Dt, 10 Lf of
`Tt, 25 ug of Pt, 25 wg of FHA, 40 DU (Antigen D Unit) of
`IPV1,8 DU ofIPV2, 32 DU ofIPV3, 12 ug ofPRP (in PRP-Tt
`
`conjugate form), 0.6 mg ofAl, 55 mM ofphosphate ions, 20
`mMof carbonale ions, and Tris sucrose buffer, 2.5 mM,
`2.125%, at pH 6.8-7.2.
`[0062] All the syringes of the three categories were stored
`horizontally at 25° C. for two months (accelerated ageing).
`The HBsAg desorption was measured in each of the three
`categories at T=0 (just after loading the syringes) and. then
`after two months.
`
`[0063] Desorption was evaluated by centrifuging, the con-
`tent of the syringes and then measuring the amountof des-
`orbed HBsAgpresent in the supernatant by ELISA (sandwich
`ELISA,involving a mouse anti-HBsAg monoclonal antibody
`(gM)for coating and capture, a second mouse anti-HBsAg
`monoclonal antibody (IgG) and a third anti-mouse IgG poly-
`clonal antibody coupled to peroxydase (Sigma, Ref. A3673)
`whichis revealed by adding tetramethyl benzidine).
`[0064] AtT (timce)=0, the HbsAg adsorption level was iden-
`tical in the three categories (98% of the total HBsAg was
`adsorbed). At T—2 months, desorption was observedinall the
`categories, but the desorption percentage differed depending
`upon the category. The highest desorption percentage was
`found in category (1) (At T—1 and 2 months, 55 and 50% of
`the total HBsAg was adsorbed, respectively), whereas the
`lowest percentage was found in category (3) (At T=1 and 2
`months, 72 and 69% of the total HBsAg was adsorbed,
`respectively).
`[0065]
`B—Abulk of the vaccine composition described in
`A—was distributed into two categories of single-dose 1 mL
`syringes, the characteristics of which were as follows:
`[0066]
`(1) standard siliconized glass syringes (free sili-
`cone); and
`[0067]
`(2) non-siliconized syringes.
`[0068] The bulk was distributed in 0.5 mL single doses,
`each dose containing 10 ug of HBsAg, 30 Lf of Dt, 10 Lf of
`Tt, 25 ug of Pt, 25 ug of FHA, 40 DU (Antigen D Unit) of
`IPV1, 8 DU of[PV2, 32 DU ofIPV3, 12 ugofPRP Gn PRP-Tt
`conjugate form), 0.6 mg ofAl, 55 mM ofphosphate ions, 20
`mMof carbonale ions, and Tris sucrose buffer, 2.5 mM,
`2.125%, at pH 6.8-7.2.
`[0069] All the syringes of the twocategories were stored
`vertically al 25° C. for two months (accelerated ageing). The
`HBsAgdesorption was measured in each ofthe two catego-
`ries at T=0 Gust after filling syringes) and then after two
`months, as described in A—above.
`[0070] AtT (time)=0, the HbsAgadsorption level was iden-
`tical in the two categories (98% of the total HBsAg was
`adsorbed). At T=2 months, desorption was observedinall the
`categories, but the desorption percentage differed depending
`upon the category. The highest desorption percentage was
`foundin category (1) (At T=1 and 2 months, 69 and 68% of
`the total HBsAg was adsorbed, respectively), whereas the
`lowest desorption percentage was found in category (2) (At
`T=1 and 2 months, 73% of the total HBsAg was adsorbed).
`
`This clearly indicates that the antigen adsorption onto an
`aluminumadjuvant is sensitive to free silicone.
`[0071]
`C—Abulk of the vaccine composition described in
`A—wasdistributed into three types of single-dose 1 mf.
`syringes, the characteristics of which were as follows:
`[0072]
`(1) lowsiliconized glass syringes (50-100 ug free
`silicone/syringe);
`[0073]
`(2) standard highly siliconized glass syringes
`((RTFsyringe Luercone™from Gerresheimer): 800 ug
`to 1 mgfree silicone/syringe); and
`
`Regeneron Exhibit 1014.005
`
`

`

`US 2012/0091026 Al
`
`Apr. 19, 2012
`
`
`
`19, The method as claimed in claim 7, wherein the antigen
`contained in the composition is hepatitis B surface antigen
`and the aluminum adjuvant is aluminum oxy hydroxide.
`20. The container as claimed in claim 16, wherein the
`container is the reservoir of a syringe that is closed by a
`plunger.
`21. The container as claimed in claim 13, wherein the
`antigen contained in the composition is hepatitis B surface
`antigen and the aluminum adjuvantis aluminum oxy hydrox-
`ide.
`
`9. A container for storing and/or administering a vaccine
`(3) syringes having inner surface coated with
`[0074]
`composition, wherein the container (i) contains a vaccine
`polymerized silicone (50-100 ug/syringe).
`composition comprising, an antigen adsorbed on an aluminum
`[0075]
`Syringes of category (1) are operative only if the
`adjuvant and (ii) is closed. by a device acting as a stopper,
`plunger used for injection is also siliconized, because the
`wherein the surface of the device contactable with the com-
`amountof silicone coating the inner surface ofthe syringeis
`positionis coated with a fluoropolymer.
`too lowto allowsliding on its own.
`10. The container as claimed in claim 9, wherein the vac-
`[0076] All the syringes (types 1 to 3) were closed with the
`same type of non-siliconized stopper.
`cine composition is in liquid or lyophilized form.
`[0077] The bulk vaccine was distributed in 0.5 mL single
`11. The container as claimed in claim 10, wherein the
`doses, each dose being as described in A and B.
`fluoropolymeris selected from the group consisting of poly-
`[0078] All the syringes of the three types were stored ver-
`tetrafluoroethylene
`(PTFE),
` polytetrafluoropropylene
`tically at 25° C. for two months (accelerated ageing). The
`(PTFP), fluorinated ethylene propylene (FEP, a copolymerof
`HBsAgdesorption was measured in each ofthe three typesat
`hexafluoropropylene andtetrafluoroethylene), polychlorotr-
`T=0 (just after filling svringes) and then after two months, as
`fluoroethylene (PCTFE), perfluoroalkoxy co-polymer (PFA),
`described in A above.
`poly(ethylene-co-tetrafluorvethylene)
`(ETFE), poly(ethyl-
`[0079] AtT (time)=0, the HbsAg adsorption level was iden-
`enechlorotrifluoroethylene)
`(ECTFE), polyvinyl
`fluoride
`tical
`in the three types (94% of the total HBsAg was
`(PVF) and polyvinylidene fluoride (PVPF).
`adsorbed). At ‘l1=2 months, desorption was observed in all
`12. A containerfor storing and/or administering a vaccine
`syringes, but the desorption percentage differed depending
`composition, wherein the container (1) has inner surfaces
`upon the type. The highest desorption percentage was found
`coated with polymerizedsilicone and (11) contains a vaccine
`in type (2) (At T—1 and 2 months, 60 and 58% of the total
`composition comprising an antigen adsorbed on an aluminum
`HBsAgwasadsorbed, respectively), whereas the desorption
`adjuvant.
`percentage was found similar in types (1) and (3) and defini-
`13, The container as claimed in claim 12, wherein the
`tively much lower than in type (2): In type (1), at T=1 and 2
`vaccine composition is in liquid or lyophilized form.
`months, 73% and 68%of the total HBsAg was adsorbed,
`14. A containerfor storing and/or administering a vaccine
`respectively. In type (3), at T=1 and 2 months, 69% and 66%
`composition, wherein the container (1) has inner surtaces
`ofthe total HBsAg wasadsorbed,respectively.
`coated with polymerized silicone;
`(11) contains a vaccine
`[0080] Again, this clearly indicates that Gi) the antigen
`composition comprising an antigen adsorbed on an aluminum
`adsorption onto an aluminum adjuvant is sensitive to tree
`adjuvant; and (iii) which is closed by a device acting as a
`silicone loaded in an amount necessary for sliding and (11)
`stopper, wherein the surface ofthe device contactable with
`polymerization ofsilicone allows overcomingthis issue.
`the composition is coated with a fluoropolymer.
`[0081]
`D—Astability stu