Europll1che1
`Patentemt
`European
`Patent Office
`Office eufl>phn
`delbnlwetl
`
`Bescheinigung
`
`Certificate
`
`Attestation
`
`Die angehefteten
`Unterlagen stimmen mit der
`als ursprOnglich eingereicht
`geltenden Fassung der auf
`dem nachsten Blatt
`bezeichneten europaischen
`Patentanmeldung Uberein.
`
`The attached documents are
`exact copies of the text in
`which the European patent
`application described on the
`following page is deemed to
`have been filed.
`
`Les documents joints a la
`presente attestation sont
`conformes au texte,
`considere comme
`initialement depose, de la
`demande de brevet
`europeen qui est specifiee a
`la page suivante.
`
`Patentanmeldung Nr.
`
`Patent application No.
`
`Demande de brevet n°
`
`12174860.2 / EP12174860
`
`The organization code and number of your priority application, to be used for filing abroad under the Paris
`Convention, is EP12174860.
`
`EPA/EPO/OEB Form 1014
`
`05.12
`
`Der Prasident des Europaischen Patentamts;
`Im Auftrag
`
`For the President of tlie European Patent Office
`
`Le President de !'Office europeen des brevets
`p.o.
`
`U. lngmann
`
`MV22832
`
`Regeneron Exhibit 1035.001
`
`

`

`Anmeldung Nr:
`Application no.:
`Demande no:
`
`12174860.2
`
`Anmelder / Applicant(s) / Demandeur(s):
`
`Novartis AG
`Lichtstrasse 35
`4056 Basel/CH
`
`Anmeldetag:
`Date of filing:
`Date de depot:
`
`03.07.12
`
`Bezeichnung der Erfindung / Title of the invention / Titre de !'invention:
`(Falls die Bezeichnung der Erfindung nicht angegeben ist, oder falls die Anmeldung in einer Nicht-Amtssprache des EPA eingereicht
`wurde, siehe Beschreibung bezuglich ursprunglicher Bezeichnung.
`If no title is shown, or if the application has been filed in a non-EPO language, please refer to the description for the original title.
`Si aucun titre n'est indique, ou si la demande a ete deposee dens une langue autre qu'une langue officielle de l'OEB, se referer a la
`description pour le titre original.)
`
`Syringe
`
`In Anspruch genommene Prioritat(en) / Priority(Priorities) claimed/ Priorite(s) revendiquee(s)
`Staat/Tag/Aktenzeichen / State/Date/File no. / Pays/Date/Numero de depot:
`
`Am Anmeldetag benannte Vertragstaaten I Contracting States designated at date of filing / Etats contractants designees tors
`du depot:
`
`AL AT BE BG CH CY CZ DE DK EE ES Fl FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL
`PT RO RS SE SI SK SM TR
`
`EPA/EPO/OEB Form 1014
`
`05.12
`
`2
`
`Regeneron Exhibit 1035.002
`
`

`

`PAT055157-EP-EPA
`
`TECHNICAL FIELD
`
`SYRINGE
`
`The present invention relates to a syringe, particularly tu a small volume syringe such as a
`
`syringe suitable for ophthalmic injection8.
`
`5
`
`BACKGROUND ART
`
`Many medicaments are delivered to a patient in a syringe from which the user can dispense the
`
`medicament. If medicament is delivered to a patient in a syringe it is often to enable the patient,
`
`or a caregiver, to inject the medicament.
`
`It is important for patient safety and medicament
`
`integrity that the syringe and the contents of that syringe are sufficiently sterile to avoid
`
`l O
`
`infection, or other, risks for patients. Sterilisation can be achieved by terminal sterilisation in
`
`which the assembled product, typically already in its associated packaging, is sterilised using
`
`heat or a sterilising gm;.
`
`For small volume syringes, for example those for injections into the eye in which it is intended
`
`that about 0.1 ml or less of liquid is to be injected the sterilisation can pose difficulties that are
`
`15
`
`not necessarily associated with larger syringes. Changes in pressure, internal or external to the
`
`syringe, can cause parts of the syringe to move unpredictably, which may alter sealing
`
`characteristics and potentially compromise sterility. Incorrect handling of the syringe can also
`
`pose risks to product sterility.
`
`Furthermore, certain therapeutics such as biologic molecules are patiicularly sensitive to
`
`20
`
`sterilisation, be it cold gas sterilisation, thermal sterilisation, or irradiation. Thus, a careful
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`balancing act is required to ensure that while a suitable level of sterilisation is carried out, the
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`syringe remains suitably sealed, such that the therapeutic is not compromised.
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`There is therefore a need for a new syringe construct which provides a robust seal for its content,
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`but which maintains ease of use.
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`25
`
`DISCLOSURE OF THE INVENTION
`
`The present invention provides a pre-filled syringe, the syringe comprising a body, a stopper and
`
`a plunger, the body comprising an outlet at an- outlet end and the stopper being arranged within
`
`the body such that a front surface of the stopper and the body define a variable volume chamber
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`from which a fluid can be expelled though the outlet, the plunger comprising a plunger contact
`
`30
`
`surface at a first end and a rod extending between the plunger contact surface and a rear portion,
`
`-1-
`
`Regeneron Exhibit 1035.003
`
`

`

`PAT055157-EP-EPA
`
`the plunger contact surface arranged to contact the stopper, such that the plunger can be used to
`
`force the stopper towards the outlet end of the body, reducing the volume of the variable volume
`chamber, characterised in that the fluid comprises an ophthalmic solution. In one embodiment,
`the ophthalmic solution comprises a VEGF-antagonist.
`
`5
`
`In one embodiment, the syringe is suitable for ophthalmic injections, more particularly
`
`intravitreal injections, and as such has a suitably small volume. The syringe may also be silicone
`
`free, or substantially silicone free, or may comprise a low level of silicone oil as lubricant.
`
`For ophthalmic injections, it is particularly important for the ophthalmic solution to have
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`particularly low particle content. In one embodiment, the syringe meets US Pharmacopeia
`
`lO
`
`standard 789 (USP789).
`
`Syringe
`
`The body of the syringe may be a substantially cylindrical shell, or may include a substantially
`
`cylindrical bore with a non circular outer shape. The outlet end of the body includes an outlet
`through which a fluid housed within the variable volume chamber can be expelled as the volume
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`15
`
`of said chamber is reduced. The outlet may comprise a projection from the outlet end through
`
`which extends a channel having a smaller diameter than that of the variable volume chamber.
`
`The outlet may be adapted, for example via a luer lock type connection, for connection to a
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`needle or other accessory such as a sealing device which is able to seal the variable volume
`
`chamber, but can be opemte<l, or removed, to unseal the variable volume chamber an<l allow
`
`20
`
`connection of the syringe to another accessory, such as a needle. Such a connection may be
`
`made directly between the syringe and accessory, or via the sealing device. The body extends
`
`along a first a.xis from the outlet end to a rear end.
`
`The body may be made from a plastic material (e.g. a cyclic olefin polymer) or from glass and
`
`may include indicia on a surface thereof to act as an injection guide. In one embodiment the
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`25
`
`body may comprise a priming mark. This allows the physician to align a pre-determined part of
`
`the stopper (such as the tip of the front surface or one of the circumferential ribs, discussed later)
`
`with the mark, thus expelling excess ophthalmic solution and any air bubbles from the syringe.
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`The priming process ensures that an exact, pre-determined dosage is administered to the patient.
`
`The stopper may be made from rubber, silicone or other suitable resiliently deformable materiaJ.
`
`30
`
`The stopper may be substantially cylindrical and the stopper may include one or more
`
`circumferential ribs around an outer surface of the stopper, the stopper and ribs being
`
`-2-
`
`Regeneron Exhibit 1035.004
`
`

`

`PATOSS 157-EP-EPA
`
`dimensioned such that the ribs form a substantially fluid tight seal with an internal surface of the
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`syringe body. The front surface of the stopper may be any suitable shape, for example
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`substantially planar, substantially conical or of a domed shape. The rear surface of the stopper
`
`may include a substantia!Iy central recess. Such a central recess could be used to connect a
`
`5
`
`plunger to the stopper using a snap fit feature or thread connection in a known manner. The
`
`stopper may be substantially rotationally symmetric about an axis through the stopper.
`
`The plunger comprises a plunger contact surface and extending from that a rod extends from the
`
`plunger contact surface to a rear portion. The rear portion may include a user contact portion
`
`adapted to be contacted by a user during an injection event. The user contact portion may
`
`l O
`
`comprise a substantially disc shaped portion, the radius of the disc extending substantially
`
`perpendicular to the axis along which the rod extends. The user contact portion could be any
`
`suitable shape. The axis along which the rod extends may be the first axis, or may be
`
`substantially parallel with the first axis.
`
`The syringe may include a backstop arranged at a rear portion of the body. The backstop may be
`
`15
`
`removable from the syringe. If the syringe body includes terminal flanges at the end opposite the
`
`outlet end the backstop may be configured to substantially sandwich terminal flanges of the body
`
`as this prevent movement of the backstop in a direction parallel to the first axis.
`
`The rod may comprise at least one rod shoulder directed away from the outlet end and the
`
`backstop may include a backstop shoulder directed towards the outlet end to cooperate with the
`
`20
`
`rod shoulder to substantially prevent movement of the rod away from the outlet end when the
`
`backstop shoulder and rod shoulder are in contact. Restriction of the movement of the rod away
`
`from the outlet end can help to maintain sterility during terminal sterilisation operations, or other
`
`operations in which the pressure within the variable volume chamber or outside the chamber
`
`may change. During such operations any gas trapped within the variable volume chamber, or
`
`25
`
`bubbles that may form in a liquid therein, may change in volume and thereby cause the stopper
`
`to move. Movement of the stopper away from the outlet could result in the breaching of a
`
`sterility zone created by the stopper. This is particularly important for low volume syringes
`
`where there are much lower tolerances in the component sizes and less flexibility in the stopper.
`
`The term sterility zone as used herein is used to refer to the area within the syringe that is sealed
`
`30
`
`by the stopper from access from either end of the syringe. This may be the area between a seal
`
`of the stopper, for example a circumferential rib, closest to the outlet and a seal of the stopper,
`
`for example a circumferential rib, furthest from the outlet. The distance between these two seals
`
`-3-
`
`Regeneron Exhibit 1035.005
`
`

`

`PAT055157-EP-EPA
`
`defines the sterility zone of the stopper since the stopper is installed into the syringe barrel in a
`
`sterile environment.
`
`To further assist in maintaining sterility during the operations noted above the stopper may
`
`comprise at a front circumferential rib and a rear circumferential rib and those ribs may be
`
`5
`
`separated in a direction along the first axis by at least 3mm, by at least 3.5 mm, by at least
`
`3.75mm or by 4mm or more. One or more additional ribs (for example 2, 3, 4 or 5 additional
`
`ribs, or between 1-10, 2-8, 3-6 or 4-5 additional ribs) may be arranged between the front and rear
`
`ribs. ln one embodiment there arc a total of three circumferential ribs.
`
`A stopper with such an enhanced sterility zone can also provide protection for the injectable
`
`10 medicament during a terminal sterilisation process. More ribs on the stopper, or a greater
`
`distance between the front and rear ribs can reduce the potential exposure of the medicament to
`
`the sterilising agent. However, increasing the number of ribs can increase the friction between
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`the stopper and syringe body, reducing ease of use. While this may be overcome by increasing
`
`the siliconisation of the syringe, such an increase in silicone levels is particularly undesirable for
`
`15
`
`syringes for ophthalmic use.
`
`The rod shoulder may be arranged within the external diameter of the rod, or may be arranged
`
`outside the external diameter of the rod. By providing a shoulder that extends beyond the
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`_ external diameter of the rod, but still fits within the body, the shoulder can help to stabilise the
`
`movement of the rod within the body by reducing movement of the rod perpendicular to the first
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`20
`
`axis. The rod shoulder may comprise any suitable shoulder forming elements on the rod, but in
`
`one embodiment the rod shoulder comprises a substantially disc shaped portion on the rod.
`
`In one embodiment of the syringe, when arranged with the plunger contact surface in contact
`
`with the stopper and the variable volume chamber is at its intended maximum, volume there is a
`
`clearance of no more than about 2mm between the rod shoulder and backstop shoulder. In some
`
`25
`
`embodiments there is a clearance of less than about l.5 mm and in some less than about 1 mm.
`
`This distance is selected to substantially limit or prevent excessive rearward (away from the
`
`outlet end) movement of the stopper.
`
`In one embodiment the variable volume chamber has an internal diameter greater than 5mm or
`
`6mm and less than 3mm or 4mm. The internal diameter may be between 3mm and 6mm, or
`
`30
`
`between 4mm and 5mm.
`
`-4-
`
`Regeneron Exhibit 1035.006
`
`

`

`PA T055157-EP-EPA
`
`In another embodiment the syringe is dimensioned so as to have a nominal maximum fill volume
`
`of between about 0.1ml and about 1.5ml. In certain embodiments the nominal maximum fill
`
`volume is between about 0.5ml and about 1 ml. In certain embodiments the nominal maximum
`
`fi]l volume is about 0.5ml or about 1ml, or about 1.5ml.
`
`5
`
`The length of the body of the syringe may be less than 70mm, less than 60mm or less than
`
`50mm. In one embodiment the length of the.syringe body is between 45mm and 50mm.
`
`In one embodiment, the syringe is filled with between about 0.01ml and about 1.5ml (for
`
`example between about 0.05ml and about I ml, between about 0. I ml and about 0.5ml, between
`
`about 0.15ml and about 0.175ml) of a VEGF antagonist solution. In one embodiment, the
`
`10
`
`syringe is filled with 0.165ml of a VEGF antagonist solution. Of course, typically a syringe is
`
`fi11ed with more than the desired dose to be administered to the patient, to take into account
`
`wastage due to "dead space" within the syringe and needle. There may also be a certain amount
`
`of wastage when the syringe is primed by the physician, so that it is ready to inject the patient.
`
`Thus, in one embodiment, the syringe is fil1ed with a dosage volume (i.e. the volume of
`
`15 medicament intended for delivery to the patent) of between about 0.01 ml and about 1.5ml (e.g.·
`
`between about 0.05ml and about 1ml, between about 0.1ml and about 0.5ml) of a VEGF
`
`antagonist solution. In one embodiment, the dosage volume is between about 0.03ml and about
`
`0.05ml. For example, for Lucentis, the dosage volume is 0.05ml or 0.03ml (0.5mg or 0.3mg) ofa
`
`lOmg/ml injectable medicament solution; for Eylea, the dosage volume is 0.05ml of a 40mg/ml
`
`20
`
`injectable medicament solution.
`
`In one embodiment the length of the syringe body is between about 45mm and about 50mm, the
`
`internal diameter is between about 4mm and about 5mm, the fill volume is between about 0.12
`
`and about 0.3ml and the dosage volume is between about 0.03ml and about 0.05ml.
`
`As the syringe contains a medicament solution, the outlet may be reversibly sealed to maintain
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`25
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`sterility of the medicament. This sealing may be achieved through the use of a sealing device as
`
`is known in the art. For example the OVS™ system which is available from Vetter Pharma
`
`lntcmational GmbH.
`
`It is typical to siliconise the syringe in order to allow ease of use, i.e. to apply silicone to the
`
`inside of the barrel, which decreases the force required to move the stopper. However, for
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`30
`
`ophthalmic use, it is desirable to decrease the likelihood of silicone droplets being injected into
`
`the eye. Furthermore, silicone can cause proteins to aggregate. A typical I ml syringe comprises
`
`-5-
`
`Regeneron Exhibit 1035.007
`
`

`

`PAT055157-EP-EPA
`
`100-SOOµg silicone in the barrel. Thus, in one embodime!}t, a syringe according to the invention
`
`comprises less than about 800µg (i.e. about less than about 500µg, less than about 300µg, less
`
`than about 200µg, less than about lOOµg, less than about 75µg, less than about 50µg, less than
`
`about 25µg, less than about 15µg, less than about lOµg) silicone in the barrel. Methods for __
`
`5 measuring the amount of silicone in such a syringe barrel are known in the art and include, for
`
`example, differential weighing methods and quantitation by infrared-spectroscopy of the oil
`
`diluted in a suitable solvent.
`
`During testing it was found that, for syringes having small dimensions, such as those discussed
`
`above, and particularly those described in conjunction with the Figures below, the break loose
`
`lO
`
`and sliding forces for the stopper within the syringe are substantially unaffected by reducing the
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`siliconisation levels far below the current standard to the levels discussed here.
`
`In one
`
`embodiment the glide force for the stopper within the pre-filled syringe is less than about I IN or
`
`less than 9N, less than 7N, less than SN or between abo1,1t 3N to 5N. Having too great a force
`
`required to move the stopper can cause problems during use for some users, for example
`
`15
`
`accurate dose setting or smooth dose delivery may be made more difficult if significant strength
`
`is required to move, and/or keep in motion, the stopper.
`
`In one embodiment the syringe barrel has an internal coating of silicone that has an average
`
`thickness of about 450nm or less (i.e. 400nm or less, 350nm or less, 300nm or less, 200nm or
`
`less, I 00nm or less, 50nm or less, 20nm or less). Methods to measure the thickness of silicone in
`
`20
`
`a syringe are known in the art and include the rap.ID Layer Explorer® Application, which can
`
`also be used to measure the mass of silicone inside a syringe barrel.
`
`In one embodiment, the syringe is silicone free, or substantially silicone free. Such low silicone
`
`levels can be achieved by using uncoated syringe barrels and/or by avoiding the use of silicone
`
`as a lubricant for product contacting machine parts, or pumps in the syringe assembly and fill
`
`25
`
`line.
`
`The syringe according to the invention may also meet certain requirements for particulate
`
`content. ln one embodiment, the ophthalmic solution comprises no more than 2 particles .::::50µm
`
`in diameter per ml. In one embodiment, the ophthalmic solution comprises no more than 5
`
`particles :=:;25µm in diameter per ml. In one embodiment, the ophthalmic solution comprises no
`
`30 more than 50 particles .::::lOµm in diameter per ml. In one embodiment, the ophthalmic solution
`
`comprises no more than 2 particles .::::50µm in diameter per ml, no more than 5 particles .::::25µm
`in diameter per ml and no more than 50 particles .::::lOµm in diameter per ml. In one embodiment,
`
`-6-
`
`Regeneron Exhibit 1035.008
`
`

`

`-·- - -,...__"e.-__________
`
`PAT055157-EP-EPA
`
`a syringe according to the invention meets USP789.
`
`In one embodiment the syringe has low
`
`levels of silicone sufficient for the syringe to meet USP789.
`
`VEGF Antagonists
`
`Antibody VEGF antagonists
`
`5 VEGF is a well-characterised signal protein which stimulates angiogenesis. Two antibody VEGF
`
`antagonists have been approved for human use, namely ranibizumab (Lucentis®) and
`
`bcvacizumab (A vastin®).
`
`Non-Antibody VEGF antagonists
`
`[n one aspect of the invention, the non-antibody VEGF antagonist is an immunoadhesin. One
`
`10
`
`such immuoadhesin is aflibercept (Eylea®), which has recently been approved for human use
`
`and is also known as VEGF-trap (Holash et al. (2002) PNAS USA 99: 11393-98; Riely & Miller
`
`(2007) Clin Cancer Res 13:4623-?s). Atliberc<?pt is the preferred non-antibody VEGF antagonist
`
`for use with the invention. Aflibercept is a recombinant human soluble VEGF receptor fusion
`
`protein consisting of portions of human VEGF receptors I and 2 extracellular domains fused to
`
`15
`
`the Fe portion of human TgG I. Tt is a dimeric glycoprotein with a protein molecular weight of 97
`
`kilodaltons (kDa) and contains glycosylation, constituting an additional 15% of the total
`
`molecular mass, resulting in a total molecular weight of 115 kDa. It is conveniently produced as
`
`a glycoprotein by expression in recombinant CHO Kl cells. Each monomer can have the
`
`following amino acid sequence (SEQ ID NO: 1):
`
`20
`
`25
`
`SDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATY
`KEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPS
`SKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPP
`CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
`YRVVSVL TVLHQDWLNGKEYKCKVSNKALPAP I EKT IS KAKGQPRE PQVYTLPPS RDELTI-CNQVS LTCLVK
`GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL
`SLSPG
`
`and disulfide bridges can be formed between residues 30-79, 124-185, 246-306 and 352-410
`
`within each monomer, and between residues 211-211 and 214-214 between the monomers.
`
`Another non-antibody VEGF antagonist immunoa<lhesin currently in pre-clinical development is
`
`30
`
`a recombinant human soluble VEGF receptor fusion protein similar to VEGF-trap containing
`
`extracellular ligand-binding domains 3 and 4 from VEGFR2/KDR, and domain 2 from
`
`VEGFR J /Flt- I; these domains are fused to a human IgG Fe protein fragment (Li et al., 20 I l
`
`-7-
`
`Regeneron Exhibit 1035.009
`
`

`

`PAT055157-EP-EPA
`
`Molecular Vision 17:797-803). This antagonist binds to isoforms VEGF-A, VEGF-B and VEGF(cid:173)
`
`C. The molecule is prepared using two different production processes resulting in different
`
`glycosylation patterns on the final proteins. The two glycoforms are referred to as KH902
`
`(conbercept) and KH906. The fusion protein can have the following amino acid sequence (SEQ
`
`5
`
`IDN0:2):
`
`10
`
`MVSYWDTGVLLCALLSCLLLTGSSSGGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDT
`LIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEK
`LVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSG
`LMTKKNSTFVRVHEKPFVAFGSGMESLVEATVGERVRLPAKYLGYPPPEIKWYKNGIPLESNHTIKAGHVL
`TIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVYVPPGPGDKTHTCPLCPAPELLGGPSVFLFPPKPKDT
`LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
`KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK
`ATPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
`
`and, like VEGF-trap, can be present as a dimer. This fusion protein and related molecules are
`
`15
`
`further characterized in EP 1767546.
`
`Other non-antibody VEGF antagonists include antibody mimetics (e.g. Affibody@ molecules,
`
`affilins, affitins, anticalins, avimers. Kunitz domain peptides, and monobodies) with VEGF
`
`antagonist activity. This includes recombinant binding proteins comprising an ankyrin repeat
`
`domain that binds VEGF-A and prevents it from binding to VEGFR-2. One example for such a
`
`20 molecule is DARPin@) MPO 112. The ankyrin binding domain may have the following amino
`
`acid sequence (SEQ ID NO: 3):
`
`GSDLGKKLLEAARAGQDDEVRILMANGADVNTADSTGWTPLHLAVPWGHLEIVEVLLKYGADVNAKDFQGW
`TPLHLAAAIGHQEIVEVLLKNGADVNAQDKFGKTAFDISIDNGNEDLAEILQKAA
`
`Recombinant binding proteins comprising an ankyrin repeat domain that binds VEGF -A and
`
`25
`
`prevents it from binding to VEGFR-2 are described in more detail in W02010/060748 and
`
`W020I l/l35067.
`
`Further specific antibody mimetics with VEGF antagonist activity are the 40 kD pegylated
`
`anticalin PRS-050 and the monobody angiocept (CT-322).
`
`The afore-mentioned non-antibody VEGF antagonist may be modified to further improve their
`
`30
`
`pharmacokinetic properties or bioavailability. For example, a non-antibody VEGF antagonist
`
`may be chemically modified (e.g., pegylated) to extend its in vivo half-life. Alternatively or in
`
`addition, it may be modified by glycosylation or the addition of further glycosylation sites not
`
`-8-
`
`Regeneron Exhibit 1035.010
`
`

`

`PAT055157-EP-EPA
`
`present in the protein sequence of the natural protein from which the VEGF antagonist was
`
`derived.
`
`Variants of the above-specified VEGF antagonists that have improved characteristics for the
`
`desired application may be produced by the addition or deletion of amino acids. Ordinarily, these
`
`5
`
`amino acid sequence variants will have an amino acid sequence having at least 60% amino acid
`
`sequence identity with the amino acid sequences of SEQ ID NO: I, SEQ ID NO: 2 or SEQ ID
`NO: 3, preferably at lea.st 80%, more preferably at least 85%, more preferably at least 90%, and
`most preferably at least 95%, including for example, 80%, 81%. 82%, 83%, 84%, 85%, 86%,
`
`87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and 100%. Identity
`
`l O
`
`or homology with respect to this sequence is defined herein as the percentage of amino acid
`
`residues in the candidate sequence that are identical with SEQ lD NO: l, SEQ ID NO: 2 or SEQ
`
`10 NO: 3, after aligning the sequences and introducing gaps, if necessary, to achieve the
`
`maximum percent sequence identity, and not considering any conservative substitutions as part
`
`of the sequence identity.
`
`15
`
`Sequence identity can be determined by standard methods that are commonly used to compare
`
`the similarity in position of the amino acids of two polypeptides. Using a computer program
`
`such as BLAST or FAST A, two polypeptides are aligned for optimal matching of their
`
`respective amino acids ( either along the full length of one or both sequences or along a pre(cid:173)
`
`determined portion of one or both sequences). The programs provide a default opening penalty
`
`20
`
`and a default gap penalty, and a scoring matrix such as PAM 250 [a standard scoring matrix; see
`
`Dayhoff et al., in Atlas of Protein Sequence and Structure, vol. 5, supp. 3 (1978)] can be used in
`
`cmtjunction with the computer program. For example, the percent identity can then be
`
`calculated as: the total number of identical matches multiplied by 100 and then divided by the
`
`sum of the length of the longer sequence within the matched span and the number of gaps
`
`25
`
`introduced into the longer sequences in order to align the two sequences.
`
`Preferably, the non-antibody VEGF antagonist of the invention binds to VEGF via one or more
`
`protein domain(s) that are not derived from the antigen-binding domain of an antibody. The non(cid:173)
`
`antibody VEGF antagonist of the invention are preferably proteinaccous, but may include
`
`modifications that are 11011-proteinaceous (e.g., pegylation, glycosylation).
`
`30
`
`Therapy
`
`The syringe of the invention may be used to treat an ocular disease, including but not limited to
`
`choroidal neovascularisation, age-related macular degeneration (both wet and dry forms),
`
`-9-
`
`Regeneron Exhibit 1035.011
`
`

`

`PAT055157-EP-EPA
`
`macular edema secondary to retinal vein occlusion (RVO) including both branch RVO (bR VO)
`
`and central RVO (cRVO), choroidal neovascularisation secondary to pathologic myopia (PM),
`
`diabetic macular edema (DME), diabetic retinopathy, and proliferative retinopathy.
`
`Thus. the invention provides a method of treating a patient suffering from of an ocular disease
`
`5
`
`selected from choroidal neovascularisation, wet age-related macular degeneration, macular
`
`edema secondary to retinal vein occlusion (RVO) including both branch RVO (bRVO) and
`central RVO (cRVO), choroidal neovascularisation secondary to pathologic myopia (PM),
`
`diabetic macular edema (DME), diabetic rctinopathy, and proliferative rctinopathy, comprising
`
`the step of administering an ophthalmic solution to the patient using a pre-filled syringe of the
`
`lO
`
`invention. This method prefen1bly further comprises an initial priming step in which the
`
`physician depresses the plunger of the pre-filled syringe to align the pre-determined part of the
`
`stopper with the priming mark.
`
`Kits
`
`Also provided are kits comprising the pre-filled syringes of the invention. In one embodiment,
`
`15
`
`such a kit comprises a pre-filled syringe of the invention in a blister pack. The blister pack may
`
`itself be sterile on the inside. In one embodiment, syringes according to the invention may be
`
`placed inside such blister packs prior to undergoing sterilisation, for example terminal
`
`sterilisation.
`
`Such a kit may further comprise a needle for administration of the VEGF antagonist. If the
`
`20 VEGF antagonist is to be administered intravitreally, it is typical to use a 30-gauge x !h inch
`
`needle, though 31-gauge and 32-gauge needles may be used. Such kits may further comprise
`
`instructions for use. In one embodiment, the invention provides a carton containing a pre-filled
`
`syringe according to the invention contained within a blister pack, a needle and optionally
`
`instructions for administration.
`
`25
`
`Sterilisation
`
`As noted above, a terminal sterilisation process may be used to sterilise the syringe and such a
`
`process may use a known process such as an ethylene oxide or a hydrogen peroxide sterilisation
`
`process. Needles to be used with the syringe may be sterilised by the same method, as may kits
`
`according to the invention.
`
`30 The package is exposed to the sterilising gas until the outside of the syringe is sterile. Following
`
`such a process, the outer surface of the syringe may remain sterile (whilst in its blister pack) for
`
`-10-
`
`Regeneron Exhibit 1035.012
`
`

`

`PAT055157-EP-EPA
`
`up to 6 months, 9 months, 12 months, 15 months, 18 months or longer. In one embodiment, less
`
`than one syringe in a million has detectable microbial presence on the outside of the syringe after
`
`18 months of storage. In one embodiment, the pre-filled syringe has been sterilised using EtO
`with a Sterility Assurance Level of at least 10-6
`
`. Of course, it is a requirement that significant
`
`5
`
`amounts of the sterilising gas should not enter the variable volume chamber of the syringe. The
`
`term "significant amounts" as used herein refers to an amount of gas that would cause
`
`unacceptable modification of the ophthalmic solution within the variable volume chamber. Tn
`
`one embodiment, the sterilisation process causes .::::_10% (preferably .::::_5%, .::::_3%, .::::_1%) alkylation
`
`of the VEGF antagonist. In one embodiment, the pre-filled syringe has been sterilised using EtO,
`
`10
`
`but the outer surface of the syringe has .:::lppm, preferably S0.2ppm EtO residue.
`
`Ge11eral
`
`The term "comprising" means "including" as well as "consisting" e.g. a composition
`"comprising" X may consist exclusively of X or may include something additional e.g. X + Y.
`
`The term "about" in relation to a numerical value x means, for example, x±l0%.
`
`15
`
`References to a percentage sequence identity between two amino acid sequences means that,
`
`when aligned, that percentage of amino acids are the same in comparing the two sequences. This
`
`alignment and the percent homology or sequence identity can be determined using software
`
`programs known in the art, for example those described in section 7.7.18 of Current Protocols in
`
`Molecular Biolog,-1 (F.M. Ausubel et al., eds., 1987) Supplement 30. A preferred alignment is
`
`20
`
`determined by the Smith-Waterman homology search algorithm using an affine gap search with
`
`a gap open penalty of 12 and a gap extension penalty of 2, BLOSUM matrix of 62. The Smith(cid:173)
`
`Waterman homology search algorithm is disclosed in Smith & Waterman (1981) Adv. Appl.
`
`Math. 2: 482-489
`
`25
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`Figure l shows a side view of a syringe
`
`Figure 2 shows a cross section of a top down view of a syringe
`
`Figure 3 shows a view of a plunger
`
`Figure 4 shows a cross section though a plunger
`
`-11-
`
`Regeneron Exhibit 1035.013
`
`

`

`PAT055157-EP-EPA
`
`Figure 5 shows a stopper
`
`MODES FOR CARRYING OUT THE INVENTION
`
`The invention will now be further described, by way of example only, with reference to the
`
`drawings.
`
`5
`
`Figure l shows a view from a side of a syringe l comprising a body 2, plunger 4, backstop 6 and a
`
`sealing device 8.
`
`Figure 2 shows a cross section through the syringe 1 of Figure 1 from above. The syringe 1 is
`
`suitable for use in an ophthalmic injection. The syringe 1 comprises a body 2, a stopper l O and a
`
`plunger 4. The syringe l extends along a first axis A. The body 2 comprises an outlet 12 at an
`
`lO
`
`outlet end 14 and the stopper 10 is arranged