DocCode — SEQ.TXT
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`SCOREPlaceholder Sheet for IFW Content
`
`Application Number: 13750352
`
`DocumentDate: 01/25/2013
`
`The presenceofthis form in the IFW record indicates that the following document type was
`received in electronic format on the date identified above. This content is stored in the SCORE
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`SequenceListing
`
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`Form Revision Date: February 8, 2006
`
`Regeneron Exhibit 1002.0001
`
`

`

`PATO5S157-USNP —::
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`nnnnnrrewrrenevretenvntetendAAAtARAN
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`FRING BY “EXPRESS MAIL” UNDER 37 CER1.46
`oo
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`Express Mall LabalNumber
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`“Bate of Deposit
`
`Commissioner for Patents.
`PO Box 1440
`Alexandria, VA 22343-1486
`
`UTILITY PATENT APPLICATION TRANSMITTAL AND FEE SHEET
`
`Transmitted herewith for filing under 37 CFR §1.53(b) Is the utility patent application of
`
`Applicant (or identifier):
`
`Sigg, Juergen et al.
`
`Title:
`
`SYRINGE
`
`Enclosed are:
`
`ON
`
`CRIBS
`
`CIO
`MIXES
`
`Specification (including Claims and Abstract) - 27 pages
`Drawings - 1 sheets
`Executed Declaration and Power of Attorney(original or capy)
`Microfiche Computer Program (appendix)
`Nucleotide and/or Amina Acid Sequence Submission
`i} Computer Readable Copy
`{] Paper Copy
`i] Staternent Verifying identity of Above Copies
`Preliminary Amendment
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`Other. a) Submission of Sequence Listing including Statement of Verification (4
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`b) Unexecuted Declaration for Udlity Application using an Appiication Data
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`©) Transmittal for Power of Attorney to one or more registered Practitioners
`and Power of Attorney by Applicant (2 sheets)
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`Sefore calculating the filing fee, please enter the enclosed Preliminary Amendment.
`Before caiculating the filing fee, please cancel claims
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`
`Regeneron Exhibit 1002.0002
`
`

`

`
`[Basic Filing Fes TeenBe
`Search Fee
`cL
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`sevnententnntntennetnnennnteevuneene
`ExarninationFee_
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`|Multiple Dependent Claim Fee ($ ).
`i| Foreign Language Surcharge (
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`Filed
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`eeeeee
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`independent
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`Claims
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`PExira
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`S90 orfraction thereof
`Sheels
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`rounded up to a whole number)
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`oeDdLD
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`| TOTAL FILING FEE|S$ 4970
`
`
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`Sheats
`
`
`
`XI Please charge Deposit Account No. 19-0134 in the name of Novartis in the amount of
`$1970. An additional copyof this paper is enclosed. The Commissioner is hereby
`authorized to charge any additional fees under 37 CFR §1.16 and §1.17 which may be
`required in connection with this application, or credit any overpayment, to Deposit
`Account No. 19-0134 in the name of Novartis.
`
`Megse address all carrespandence to the address associated with Customer No. 003095,
`which is currently:
`
`Novartis Pharmaceuticals Corporation
`One Health Plaza, Bldg. 104
`East Hanover, NJ 07936
`
`Please direct all telephone calls fo the undersigned at the number given below, andall
`telefaxes to +1 9737818265.
`
`Novartis Pharmacauiicais Corporation
`One Health Plaza, Bldg. 101
`East Hanover, NJ 07936
`+1 8627785816
`
`Date: January 25, 2013
`
`Respectfully submitted,
`
`/ Andraw Holmes /
`
`Andrew Holmes
`Agent for Applicant
`Rag. No. 81.819
`
`
`Regeneron Exhibit 1002.0003
`
`

`

`TECHNICAL FIELD
`
`SYRINGE
`
`The present invention relates to a syringe, particularly to a small volume syringe such as a
`syringe suitable for ophthalmic injections.
`
`5
`
`BACKGROUND ART
`
`10
`
`15
`
`20
`
`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
`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 ora sterilising gas.
`
`For small volumesyringes, for example those for injections into the eye in which it is intended
`that about 0.1ml or less of liquid is to be injected the sterilisation can pose difficulties that are
`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 compromisesterility.
`Incorrect handling of the syringe can also
`poserisks to productsterility.
`
`Furthermore, certain therapeutics such as biologic molecules are particularly sensitive to
`sterilisation, be it cold gas sterilisation,
`thermal sterilisation, or irradiation. Thus, a careful
`balancing act is required to ensure that while a suitable level ofsterilisation is carried out, the
`syringe remains suitably sealed, such that the therapeutic is not compromised. Of course, the
`syringe must also remain easy to use,
`in that the force required to depress the plunger to
`administer the medicament must not be too high.
`
`25
`
`Thereis therefore a need for a new syringe construct which provides a robustseal for its content,
`
`but which maintains case of use.
`
`DISCLOSURE OF THE INVENTION
`
`The present invention providesa 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 suchthat a front surface of the stopper and the body define a variable volume chamber
`
`30
`
`-1-
`
`
`Regeneron Exhibit 1002.0004
`
`

`

`from which a fluid can be expelled though the outlet, the plunger comprising a plunger contact
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`surface at a first end and a rod extending between the plunger contact surface and a rear portion,
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`ihe phinger 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
`
`§
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`chamber, characterised in that the fluid comprises an ophthalmic solution. In one embodiment,
`
`the ophthalmic solution comprises a VEGF-antagonist.
`
`In one embodiment,
`
`the syringe is suitable for ophthalmic injections, siore particularly
`
`intravitreal injections, and as sack has a suitably small volume, The syringe may also be silicone
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`oil free, or substantially silicone oil free, or may comprise a lowlevel of silicone oi as lubricant.
`
`1G
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`In one embodiment, despite the lowsilicone off level, the stopper break loose and slide force is
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`less than 20N.
`
`For ophthalmic iniections,
`l
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`it
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`is
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`particularly important for the ophthalmic solution to have
`5
`I
`I
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`particularly low particle content.
`
`In one embodiment,
`
`the syringe meets US Pharmacopeia
`
`standard 789 (USP789).
`
`iS
`
`Syringe
`
`The bady of the syringe may be a substantially cylindrical shell, or may include a substantially
`
`eylindrical bore with a non circular outer shape. The outlet end of the body inchides an outlet
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`through which a fluid housed within the variable volume chamber can be expelled as the volume
`of said chamberis reduced. The outlet may comprise a projection from the outlet end through
`
`20
`
`which extends a channel having a smaller diameter than that of the variable volume chamber.
`fey
`The outlet may be adapted, for example via a luer lock type connection, for connection to a
`
`needie or other accessory such as a sealing device which is able fo seal the variable volume
`
`chamber, but can be operated, or removed,
`
`to unseal the variable volume chamber and allow
`
`connection of the syringe to another accessory, such as a needle. Such a connection may be
`
`28
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`made directly between the syringe and accessory, or via the sealing device. The body extends
`
`along afirst axis from the outlet end to a rear end.
`
`The body may be made from a plastic material (e.g. a cyclic clefin polymer) or from glass and
`
`may include indicia on a surface thereof to act as an infection guide. In one embodiment the
`body may comprise a priming mark. This allows the physician to align a pre-determined part of
`the stopper (suchas the tip of the front surface or one of the circumferential ribs, discussed later)
`or plunger with the mark, thus expelling excess ophthalmic solution and any air bubbles from the
`
`30
`
`
`Regeneron Exhibit 1002.0005
`
`

`

`syringe. The priming process ensures that an exact, pre-determined dosage is administered to the
`
`patient.
`
`The stopper may be made fromrubber, silicone or other suitable resitiently deformable material.
`
`The stopper may be substantially cylindrical and the stopper may include one or more
`
`sn
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`circumferential
`
`ribs around an outer surface of the stopper,
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`the stopper and ribs being
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`dimensioned such that the ribs form a substantially fluid tight seal with an internal surface ofthe
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`syringe body. The front surface of the stopper may be any suitable shape, for example
`
`substantially planar, substantially conical or of a domed shane. The rear surface of the stopper
`
`may include a substantially central recess. Such a central recess could be used to connect a
`10 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 inclade a user contact portion
`
`adapted to be contacted by a user during an injection event. The user contact portion may
`
`13
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`comiprise a substantially disc shaped portion, the radius of the disc extending substantially
`
`perpendicular ta 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
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`substantially parallel with the first axis.
`
`The syringe may include a backstop arranged at a rear portion of the body. The backstop may be
`
`20
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`removable from the syringe. Hthe syringe body includes terminal flanges at the end opposite the
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`outlet end the backstop may be configured to substantially sandwich terminal flanges ofthe body
`
`
`
`
`
`
`
`
`
`
`
`
`
`
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`as this Heltotheth prevent movementt t of theof the backstbackstop in a direction parallel to thedirec first axis.first ,
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`
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`The rod may comprise at least one rod shoulder directed away from the outlet end and the
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`backstop may inchide a backstop shoulder directed towards the outlet end ta cooperate with the
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`25
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`rod shoulder to substantially prevent movement of the rod away fromthe outlet end when the
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`backstop shoulder and rod shoulder are in coniact. Restriction of the movement of the rod away
`
`fromthe outlet end can help io maintain sterility during termina! 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
`
`30
`
`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 im the breaching of a
`
`sterility zone created by the stopper. This is particularly important for low volume syringes
`
`63
`
`
`Regeneron Exhibit 1002.0006
`
`

`

`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
`
`bythe stopper from access from ether end of the syringe. This may be the area betweena seal
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`of the stopper, for example a circumferential rid, closest to the outlet and a seal of the stopper,
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`Hs
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`for example a circumferential rib, furthest from the outlet. The distance between these two seals
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`efines the sterility zone of the stopper since the stopper is installedinto the syringe barrel in a
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`sterile environment.
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`To further assist in maintaming sterility daring the operations noted above the stopper may
`
`comprise at a front circumferential rib and a rear circumferential rib and those ribs may be
`
`10
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`separated in a direction along the first axis by at least 3mm, by at least 3.3 mm, byat least
`
`3.75mm or by 4mm or more. One or more additional ribs (for example 2, 3, 4 or 3 additional
`
`ribs, or between 1-10, 2-8, 3-6 or 4-5 additional ribs) may be arranged between the front and rear
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`ribs.
`
`In one embodiment there are a total of three circunrferential ribs.
`
`A stopper with such an enhanced sterility zone can also provide protection for the injectable
`
`pa As
`
`medicament during a termmal sterilisation pracess. More ribs on the stopper, or a greater
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`distance between the front and rear ribs can reduce the potential exposure of the medicament tc
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`the sterilising agent. However, Increasing the number of ribs can increase the friction between
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`the stopper and syringe body, redhicing ease of use. While this may be overcome by increasing
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`the siliconisation of the syringe, such an increase insilicone off levels is particularly undesirable
`
`20
`
`for syringes for ophthalmic use.
`
`The rod shoulder may be arranged within the external diameter ofthe 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 stil fits within the body, the shoulder can help to stabilise the
`
`movement of the rod within the body by reducingmovement of the rod perpendicular to the first
`
`25
`
`axis. The rod shoulder may comprise any suitable shoulder forrning 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 chamberis at its intended maximumvolume there is a
`
`clearance of no more than about 2mm between the rod shoulder and backstop shoulder,
`
`In same
`
`30
`
`embodiments there is a clearance of less than about 1.5 mm and in some fess than about Imm.
`
`This distance is selected to substantially Hmit or prevent excessive rearward (away from the
`
`outlet end} movement ofthe stopper.
`
`
`Regeneron Exhibit 1002.0007
`
`

`

`In one embodiment the variable volume chamber has an interna! diameter greater than Smmi or
`
`6mm, or less than 3mm or 4mm. The internal diameter may be between Jram and Omm, or
`
`between 4mm and Smrn.
`
`In another embodiment the syringe is dimensioned so as to have a nominal maximumfll volume
`
`§
`
`of between about O.Im! and about {.Sml. In ceriain embodiments the nominal maximem fill
`
`yalume is between about O.$mi and about Iml.
`
`in certain embodiments the nominal maximum
`
`fill volume is about 0.5m] or about imi, or about 1.45mi.
`
`The length of the body ofthe syringe may be less than 7Omm, less than 60mm orless than
`
`50mm.
`
`In one embodiment the length of the syringe bodyis between 45mmand 50min,
`
`10
`
`In one embodiment, the syringe is filed with between about G.0iml and about 1Sml (for
`
`example between about 0.68mi and about Iml, between about O.im! and about 0.3ml, between
`
`about 0.1 Simi and about 0.17Sml} of a VEGF antagonist solution.
`
`In one embodiment,
`
`the
`
`syringe is filled with O.165mi of a VEGF antagonist solution. OF course, typically a syringe is
`
`Aled with more than the desired dose to be administered to the patient, to take into account
`
`Ht
`
`wastage due to “dead space” withinthe syringe and needle. There mayalso be a certain amount
`
`of wastage when the syringe is primed by the ohysician, so that it is ready to inject the patient.
`
`Thus,
`
`in one embodiment,
`
`the syringe is filled with a dosage yolume (Le.
`
`the volume of
`
`medicament intended for delivery to the patent) of between about 0.0iml and about 1.5m (e.g.
`
`between about 6.05mi and about Ind, between about O.Imi and about G4m) of a VEGF
`
`20
`
`antagonist solution. In one embodiment, the dosage volume is between about 0.05mi andabout
`
`G.0Sral. For example, for Lucentis, the dosage volume is 0.05mi or G.03ml (0.5mg or O.3mig) ofa
`
`lOmg/ml injectable medicament solution; for Eylea, the dosage volume is 0.0Sml of a 4Omeg/ml
`
`injectable medicament solution. Although unapproved for ophthalmic indications, bevacizumab
`
`is used off-label in such ophthalmic indications at a concentration of 25meg/ml; typically at a
`
`na a
`
`dosage volume of 6.05mi (1. 28mg). In one embodiment, the extractable volume from the syringe
`
`(that is the amount of product obtainable fromthe syringe following filling, taking Into account
`
`loss due to dead space in the syringes and needle) is about 0.09m1.
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`tn one embodiment the length of the syringe bodyis between about 43mm and about S0hmm, the
`
`internal diameter is between about 4rnm and about Smm, the fill vohume is befween about 0.12
`
`30
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`and about 0.3m! and the dosage volumeis between about 0.03mi and about 0.0SmL.
`
`
`Regeneron Exhibit 1002.0008
`
`

`

`As the syringe contains a medicament solution, the outlet may be reversibly sealed to maintain
`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
`
`International GmbH.
`
`5S
`
`It is typical to siliconise the syringe in order to allow ease of use, i.e. fo applysilicone oil to the
`inside of the barrel, which decreases the force required to move the stopper. However, for
`
`ophthalmic use, it is desirable to decrease the likeHhood ofsilicone oil droplets being injected
`into the eye. With multiple injections, the amount ofsilicone droplets can build up in the eye,
`
`causing potential adverse effects, including “floaters” and an increase im intra-ocular pressure.
`
`10
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`Furthermore, silicone oil can cause protems to aggregate. A typical ml syringe comprises 100-
`
`800p¢ silicone olf in the barrel, though a survey of manufacturers reported that 500-1000u¢ was
`
`typically used in pre-filled syringes (Badkar ef af. 2011, AAPS PharmaSciTech, 12(2):564-372).
`
`Thus, in one embodiment, a syringe according to the invention comprises less than about 800ug
`
`(Le. about less than about S0Oug, less than about 300g, Jess than about 200g, less than about
`
`15
`
`$0ug, fess than about 75pg, less than about 50pg, less than about 25p¢, less than about [Sug,
`less than about 10ug) silicone ofl in the barrel. [f the syringe comprises a lowlevel ofsilicone
`
`oil, this may be more than about lug, more than about 3ug, more than about Sug, more than
`
`about 7ug or more than about 1Oug silicone oil in the barrel. Thus,
`
`in one embodiment, the
`
`syringe may comprise about lug-about 500ug, about 3ug-about 200ug, about Spg-about O0ug
`
`20
`
`or about 10gg-about SOug silicone oil
`
`in the barrel, Methads for measuring the amount of
`
`silicone oil in such a syringe barrel are knownin the art and include, for example, differential
`
`weighing methods and quantitation by infrared-spectroscopy of the oil diluted in a suitable
`solvent. Various types of silicone ofl are available, but typically esther DC360 (Dow Corning”:
`with a viscosity of 1000cP) or DC364 emulsion (Dow Corning”; ENT360 olf with a viscosity of
` 3S80cP) are used for syringe siliconisation. In one embodiment, the pre-filled syringe of the
`
`25
`
`invention cornprises DC365 emalsion.
`
`Doring testing it was surprisingly 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 and sliding forces for the stopper within the syringe are substantially unaffected by
`
`30
`
`reducing the siliconisation levels far belowthe current standard fo the levels discussed here. This
`
`is in contrast to conventional thinking that would suggest that If you decrease the silicone ol!
`
`level, the forces required would increase (ses e.g, Schoenknecht. AAPS National Biotechnology
`Conference 2007 — Abstract no. NBC07-000488, which indicates that while 400u¢silicone ail is
`
`&-
`
`
`Regeneron Exhibit 1002.0009
`
`

`

`acceptable, usability improves when increased to 800yg). Having too great a force required to
`move the stopper can cause problems during use for some users, for example accurate dose
`setting or smooth dose delivery may be made moredifficult if significant strength is required to
`move, and/or keep in motion, the stopper. Smooth administration is particularly important in
`sensitive tissues such as the eye, where movement of the syringe during administration could
`cause local tissue damage. Break loose and slide forces for pre-filled syringes known in the art
`are typically in the region of less than 20N, but where the pre-filled syringes contain about
`100pg-about 800gsilicone oil. In one embodimentthe glide/slide force for the stopper within
`the pre-filled syringe is less than about 11N or less than 9N, less than 7N,
`less than 5N or
`between about 3N to 5N.In one embodiment, the break loose force is less than about 11N orless
`than 9N,less than 7N,less than 5N or between about 2N to 5N. Note that such measurements are
`
`for a filled syringe, rather than an empty syringe. The forces are typically measured at a stopper
`travelling speed of 190mm/min. In one embodiment, the forces are measured with a 30G x 0.5
`inch needle attached to the syringe. In one embodiment, the syringe has a nominal maximalfill
`volume of between about 0.5ml and 1ml, contains less than about 100g silicone oil and has a
`
`5
`
`10
`
`15
`
`break loose force between about 2N to 5N.
`
`In one embodimentthe syringe barrel has an internal coating of silicone oil that has an average
`thickness of about 450nm orless (i.e. 400nm or less, 350nm or less, 300nm or less, 200nm or
`less, 100nm or less, 50nm or less, 20nm orless). Methods to measure the thicknessofsilicone oil
`
`20
`
`ina syringe are knownin theart and include the rap.1D Layer Explorer® Application, which can
`
`also be used to measure the massofsilicone oil inside a syringe barrel.
`
`In one embodiment, the syringe is silicone oil free, or substantially silicone oil free. Such low
`silicone oil levels can be achieved by using uncoated syringe barrels and/or by avoiding the use
`of silicone oil as a lubricant for product contacting machine parts, or pumps in the syringe
`assembly andfill line. A further way to reducesilicone oil and inorganic silica levels in a pre-
`filled syringe is to avoid the use ofsilicone tubing in filling lines, for example between storage
`
`tanks and pumps.
`
`The syringe according to the invention may also meet certain requirements for particulate
`content. In one embodiment, the ophthalmic solution comprises no more than 2 particles >50um
`in diameter per ml. In one embodiment, the ophthalmic solution comprises no more than 5
`particles >25um in diameter per ml. In one embodiment, the ophthalmic solution comprises no
`more than 50 particles >10m in diameter per ml. In one embodiment, the ophthalmic solution
`
`25
`
`30
`
`-7-
`
`
`Regeneron Exhibit 1002.0010
`
`

`

`comprises no more than 2 particles >50um in diameter per ml, no more than 5 particles >25um
`
`in diameter per ml and no more than 50 particles >10um in diameter per ml. In one embodiment,
`
`a syringe according to the invention meets USP789 (United States Pharmacopoeia: Particulate
`
`Matter in Ophthalmic Solutions).
`
`In one embodiment the syringe has low levels of silicone oil
`
`5
`
`sufficient for the syringe to meet USP789.
`
`VEGFAntagonists
`
`Antibody VEGF antagonists
`
`VEGFis a well-characterised signal protein which stimulates angiogenesis. Two antibody VEGF
`
`antagonists have been approved for human use, namely ranibizumab (Lucentis®) and
`
`10
`
`bevacizumab (Avastin®).
`
`Non-Antibody VEGF antagonists
`
`In one aspect of the invention, the non-antibody VEGF antagonist is an immunoadhesin. One
`
`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
`
`15.
`
`(2007) Clin Cancer Res 13:4623-7s). Aflibercept 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 1 and 2 extracellular domains fused to
`
`the Fc portion of human IgG1. It is a dimeric glycoprotein with a protein molecular weight of 97
`
`kilodaltons (kDa) and contains glycosylation, constituting an additional 15% of the total
`
`20 molecular mass, resulting in a total molecular weight of 115 kDa.It is conveniently produced as
`
`a glycoprotein by expression in recombinant CHO K1 cells. Each monomer can have the
`
`following amino acid sequence (SEQ ID NO:1):
`
`SDTGRPFVEMYSEI PEI IHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATY
`
`KEIGLLTCEATVNGHLYKTNYLTHROTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPS
`
`25
`
`SKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDOGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPP
`
`CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
`
`YRVVSVLUTVLHODWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNOVSLTCLVK
`
`GFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQOGNVF SCSVMHEALHNHYTQKSL
`
`SLSPG
`
`
`
`30
`
`~=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.
`
`Regeneron Exhibit 1002.0011
`
`

`

`Another non-antibody VEGF antagonist immumoadhesin currently in pre-clinical development is
`
`a recombinant human soluble VEGF receptor fusion protein similar to YVEGF-trap containing
`
`extracellular
`
`ligand-binding domains 3 and 4 from VEGFR2/KDR, and domain 2 from
`
`VEGFRI/Flt-L; these domains are fused to a human IgG Fe protein fragment (Li et al, 2011
`
`§ Molecular Vision 17:797-803). This antagonist binds to isoforms VEGF-A, VEGF-B and VEGP-
`
`C. The molecule is prepared using two different production processes resulting in different
`
`glycosylation patterns on the final proteins, The two ghycoforms are referred to as KH9Q2
`
`conbercept) and KH906. The fasion protein can have the following ammo acid sequence (SEQ
`
`o f
`
`1D NO:2):
`
`16
`
`MYSYWDEGVLLCALLSCLLLPGSSSGGRPFVEMY SEI PRI THMTEGRELVI PCRVTSPNITVILERFELDT
`
`LIPDGKRI IWDSREGF I I SNATYREIGLLTCRATVNGHLYKTINYLUBROTNTI LDVVLSPSHG [ ELSVGER
`
`LYLNCTARTELNVGIDENWEYPS SKRAGHKKLVNRDLRTOSGS EMERF LSTLPIDGVTRSDOGLYTCAASSG
`
`LMTRANS PEVRVHERPPVAFPGSGMESLVEATVGERVRIPARY LOGY PPPEIRWYKNGT PLESNHTIRKAGEVL
`
`TIMEVSERDTGNYTVILTNPT SKERQSRVVSLVVYVEPSPSDRTHTCPLC PAPELLGGPSVFLEPPERPRD?
`
`is
`
`LMISREPEVTCVVVDV SHEDPEVKPNWYVIG VEVHNARTRPREEOYNSTYRVVSVLTVLEQDWLNGREYRCE
`
`KVSNRALPAPIBETT SKARGOPRE POVYTLEPSRDELTENOVSLTCOVRGPY PSDIAVEWESNGGOPENNYE
`
`ATPPVLDSDG SEFLYSRIPVDESRWQQGNVE SCSVMHEALHNAY TORSLSLSPG
`
`and, like VEGF-trap, can be present as a dimer. This fusion protein and related molecules are
`
`further characterized in EP1767546.
`
`20
`
`Other non-antibody VEGF antagonists include antibody mimetics (e.g. Affibody® molecules,
`
`affilins, affitins, anticalins, avimers, Kunitz domain peptides, and monabodies} with VEGF
`
`antagonist activity. This includes recombinant binding proteins comprising an ankyrin repeat
`
`domain that binds VEGF-A and prevents it from binding to VEGPR-2. One example for such a
`
`molecule is DARPin® MPOL12. The ankyrin binding domain may have the following amino
`
`oS tay
`
`acid sequence (SEQ TD NO: 3):
`
`OSDLGKELLEAARAGGDDEVRI LMANGADVNTADSTGWT PLHLAVPUGHRLEI VEVIURY GADVNARDFOQGW
`
`
`TPLHLAAATGHORIVEVLLRENGADVNAGDEFGRTAFDIS IDNGNEDLARBTLORAA
`
`Recombinant binding proteins comprising an ankyrin repeat domain that binds VEGF-A and
`
`prevents it from binding to VEGFR-2 are described in more detail in WO20104)60748 and
`
`30 WO2011/935067.
`
`Further specific antibody mimeties with VEGF antagonist activity are the 40 kD pegylated
`
`anticalin PRS-050 and the monobody angiocept (CT-322).
`
`
`Regeneron Exhibit 1002.0012
`
`

`

`The afore-mentioned non-antibody VEGF antagonist may be modified to further improve their
`
`pharmacokinetic properties or bioavailability. For example, a non-antibody VEGF antagonist
`
`may be chemically modified (e.g., pegylated} to extend its iv vive half-life. Alternatively or in
`
`addition, it may be modified by glycosylation or the addition of further glycosylation sites not
`
`§
`
`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 inyproved characteristics for the
`
`desired application may be produced bythe addition or deletion of amino acids. Ordinarily, these
`
`amino acid sequence variants will have an amino acid sequence_— at least 60% amine acid
`
`sequenceidentity with the amino acid sequences of SEG TD
`
`NOh 1, SEQ TID NO: 2 or SEQID
`
`16
`
`NO: 3, preferably at least 86%, more preferably at least 85%, more preferablyat least 90%, and
`
`mast preferably at least 95%, Including for example, 80%, 81%, 82%, 83%, 84%, 85%, 86%,
`
`87%, 88%, 89%, 90%, 91%, 92%, 93%,
`
`94%, 95%, 95%, 97%, 98%, 99%, and 140%. Tdentity
`
`or homology with respect to this sequence is defined herein as the percentage of amino acid
`
`{5
`
`residues in the candidate sequence that are identical with SEG E> NO: 1, SEQ JD NO: 2 or SEQ
`
`1D NO: 3, after aligning the sequences and introdhicing gaps, if necessary,
`
`to achieve the
`
`maximumpercent sequenceidentity, and not considering any conservative substitutions as part
`
`of the sequence identity.
`
`Seqnence identity can be determined by standard methods that are commonly used to compare
`
`20
`
`the similarity in position of the amino acids of two polypeptides. Using a computer program
`
`such as BLAST or FASTA,
`
`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-
`
`determined portion of one or both sequences}, The programs provide a default opening penalty
`and 3 default gap penalty, and a scoring matrix such as PAM 250ta standard scoring matrix; see
` Bavhoff et al., in Atlas of Protein Sequence and Structure, vol, 5, supp. 3 (19783) can be used in
`
`28
`
`confmction with the computer program.
`
`For example.
`
`the percent
`
`identity can then be
`
`caleulated as: the total mumber of identical matches multiplied by I{X) and then divided by the
`
`sum of the length of the longer sequence within the matched span and the number of gaps
`
`introduced into the longer sequences in order to align the two sequences.
`
`30
`
`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-
`
`
`Regeneron Exhibit 1002.0013
`
`Ate
`
`

`

`antibody VEGF antagonist of the invention are preferably protemaceous, but may inchide
`
`modifications that are non-proteinaceous (e.g., pegylation, glycosylation).
`
`Therapy
`
`The syringe of the invention may be used to treat an acular disease, inchuling bat not limited to
`
`het
`
`choroidal neovascularisation, age-related macular degeneration (both wet and dry forms),
`
`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 retinopathy, and proliferative retinopathy.
`
`Thus the invention provides a method of treating a patient suffering from of an ocular disease
`
`10
`
`selected from choroidal neovascularisation, wet age-related macular degeneration, macular
`
`edema secondary to retinal vein occlusion (RVO) including both branch RYO (RVO) and
`
`central RVO {(cRVO), choroidal neovascularisation secondary to pathologic pryopia (PM),
`
`diabetic macular edema (DME), diabetic retinopathy, and proliferative retinopathy, comprising
`
`the step of administering an ophthalmic solution to the patient using a pre-filled syringe of the
`
`1S
`
`invention. This method preferably 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.
`
`In one embodiment, the invention provides a method of treating an ocular disease selected from
`
`choroidal neovascu