(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2014/0012227 A1
`Sigg et al.
`(43) Pub. Date:
`Jan. 9, 2014
`
`US 201400 12227A1
`
`(54) SYRINGE
`
`(71) Applicant: NOVARTIS AG, Basel (CH)
`(72) Inventors: Juergen Sigg, Loerrach (DE),
`Christopher Royer, Munich (DE);
`Andrew Mark Bryant, Reinach (CH):
`Heinrich Martin Buettgen, Rheinfelden
`(CH); Marie Picci, Ranspack-le-bas
`(FR)
`(73) Assignee: Novartis AG, Basel (CH)
`
`Nov. 16, 2012 (DE) ......................... 2O2O12O11016.O
`Nov. 23, 2012 (DE).
`... 202O12O11259.7
`Nov. 23, 2012 (DE) ......................... 2O2O12O1126O.O
`Dec. 3, 2012 (EP) .................................. 1219536O.8
`Jan. 23, 2013 (AU).
`... 20131 OOO70
`Jan. 23, 2013 (AU) ................................ 20131 OOO71
`
`Publication Classification
`
`(51) Int. Cl.
`A6DF 9/00
`A6M5/00
`A6M5/35
`
`(2006.01)
`(2006.01)
`(2006.01)
`
`1-1.
`(22) Filed:
`Jan. 25, 2013
`(30)
`Foreign Application Priority Data
`Jul. 3, 2012 (EP).
`... 121748602
`Oct. 23, 2012 (EP).
`... 1218964.92
`Nov. 16, 2012 (AU).
`... 2012.101677
`Nov. 16, 2012 (AU).
`... 2012.101678
`
`CPC .......... A61F 9/0008 (2013.01); A61M 5/31505
`(2013.01); A61M 5/002 (2013.01)
`USPC ............................ 604/506; 604/218: 206/438
`ABSTRACT
`(7)
`The present invention relates to a syringe, particularly to a
`Small Volume Syringe such as a syringe Suitable for oph
`thalmic injections.
`
`
`
`
`
`
`
`
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`Novartis Exhibit 2044.001
`Regeneron v. Novartis, IPR2020-01318
`
`

`

`Patent Application Publication
`
`Jan. 9, 2014
`
`US 2014/0012227 A1
`
`
`
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`Novartis Exhibit 2044.002
`Regeneron v. Novartis, IPR2020-01318
`
`

`

`US 2014/0012227 A1
`
`Jan. 9, 2014
`
`SYRINGE
`
`TECHNICAL FIELD
`0001. The present invention relates to a syringe, particu
`larly to a small Volume syringe Such as a syringe Suitable for
`ophthalmic injections.
`
`BACKGROUND ART
`0002 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. Sterili
`sation can be achieved by terminal sterilisation in which the
`assembled product, typically already in its associated pack
`aging, is sterilised using heat or a sterilising gas.
`0003 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 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 com
`promise sterility. Incorrect handling of the Syringe can also
`pose risks to product sterility.
`0004 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 actis required to ensure that while a suitable
`level of sterilisation is carried out, the Syringe remains Suit
`ably 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 medi
`cament must not be too high.
`0005. There is therefore a need for a new syringe construct
`which provides a robust seal for its content, but which main
`tains ease of use.
`
`DISCLOSURE OF THE INVENTION
`0006. 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 from
`which a fluid can be expelled though the outlet, the plunger
`comprising a plunger contact surface at a first end and a rod
`extending between the plunger contact surface and a rear
`portion, 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 com
`prises an ophthalmic Solution. In one embodiment, the oph
`thalmic Solution comprises a VEGF-antagonist.
`0007. In one embodiment, the syringe is suitable for oph
`thalmic injections, more particularly intravitreal injections,
`and as such has a suitably small Volume. The Syringe may also
`be silicone oil free, or substantially silicone oil free, or may
`comprise a low level of silicone oil as lubricant. In one
`embodiment, despite the low silicone oil level, the stopper
`break loose and slide force is less than 20N.
`
`0008 For ophthalmic injections, it is particularly impor
`tant for the ophthalmic solution to have particularly low par
`ticle content. In one embodiment, the syringe meets US Phar
`macopeia standard 789 (USP789).
`Syringe
`0009. 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 of
`said chamber is reduced. The outlet may comprise a projec
`tion 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 needle or other
`accessory Such as a sealing device which is able to 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 made directly between the Syringe and
`accessory, or via the sealing device. The body extends along
`a first axis from the outlet end to a rear end.
`0010. 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 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) or plunger with the
`mark, thus expelling excess ophthalmic solution and any air
`bubbles from the Syringe. The priming process ensures that an
`exact, pre-determined dosage is administered to the patient.
`0011. The stopper may be made from rubber, silicone or
`other suitable resiliently deformable material. 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 dimensioned Such that
`the ribs form a substantially fluid tight seal with an internal
`surface of the syringe body. The front surface of the stopper
`may be any Suitable shape, for example Substantially planar,
`substantially conical or of a domed shape. The rear surface of
`the stopper may include a Substantially central recess. Such a
`central recess could be used to connect a 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.
`0012. The plunger comprises a plunger contact Surface
`and extending from that a rod extends from the plunger con
`tact 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 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 Suit
`able shape. The axis along which the rod extends may be the
`first axis, or may be substantially parallel with the first axis.
`0013 The syringe may include a backstop arranged at a
`rear portion of the body. The backstop may be 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.
`
`Novartis Exhibit 2044.003
`Regeneron v. Novartis, IPR2020-01318
`
`

`

`US 2014/0012227 A1
`
`Jan. 9, 2014
`
`0014. 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 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 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 termsterility Zone as used herein
`is used to refer to the area within the syringe that is sealed 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 defines the ste
`rility Zone of the stopper since the stopper is installed into the
`Syringe barrel in a sterile environment.
`0015 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 separated in a direction along the first axis by at
`least 3 mm, by at least 3.5 mm, by at least 3.75 mm or by 4 mm
`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. In one
`embodiment there are a total of three circumferential ribs.
`0016 A stopper with such an enhanced sterility Zone can
`also provide protection for the injectable 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 the stopper and Syringe body, reducing ease
`of use. While this may be overcome by increasing the sili
`conisation of the Syringe. Such an increase in silicone oil
`levels is particularly undesirable for syringes for ophthalmic
`SC.
`0017. The rod shoulder may be arranged within the exter
`nal diameter of the rod, or may be arranged outside the exter
`nal diameter of the rod. By providing a shoulder that extends
`beyond the 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 per
`pendicular to the first 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.
`0018. 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 2 mm
`between the rod shoulder and backstop shoulder. In some
`embodiments there is a clearance of less than about 1.5 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.
`0019. In one embodiment the variable volume chamber
`has an internal diameter greater than 5 mm or 6 mm, or less
`than 3 mm or 4 mm. The internal diameter may be between 3
`mm and 6 mm, or between 4 mm and 5 mm.
`0020. In another embodiment the syringe is dimensioned
`So as to have a nominal maximum fill Volume of between
`about 0.1 ml and about 1.5 ml. In certain embodiments the
`nominal maximum fill volume is between about 0.5 ml and
`about 1 ml. In certain embodiments the nominal maximum fill
`Volume is about 0.5 ml or about 1 ml, or about 1.5 ml.
`0021. The length of the body of the syringe may be less
`than 70 mm, less than 60 mm or less than 50 mm. In one
`embodiment the length of the syringe body is between 45 mm
`and 50 mm.
`0022. In one embodiment, the syringe is filled with
`between about 0.01 ml and about 1.5 ml (for example
`between about 0.05 ml and about 1 ml, between about 0.1 ml
`and about 0.5 ml, between about 0.15 ml and about 0.175 ml)
`of a VEGF antagonist solution. In one embodiment, the
`syringe is filled with 0.165 ml of a VEGFantagonist solution.
`Of course, typically a syringe is filled 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.
`0023 Thus, in one embodiment, the syringe is filled with
`a dosage Volume (i.e. the Volume of medicament intended for
`delivery to the patent) of between about 0.01 ml and about 1.5
`ml (e.g. between about 0.05 ml and about 1 ml, between about
`0.1 ml and about 0.5 ml) of a VEGF antagonist solution. In
`one embodiment, the dosage volume is between about 0.03
`ml and about 0.05 ml. For example, for Lucentis, the dosage
`volume is 0.05 ml or 0.03 ml (0.5 mg or 0.3 mg) of a 10 mg/ml
`injectable medicament Solution; for Eylea, the dosage Volume
`is 0.05 ml of a 40 mg/ml injectable medicament solution.
`Although unapproved for ophthalmic indications, bevaci
`Zumab is used off-label in Such ophthalmic indications at a
`concentration of 25 mg/ml; typically at a dosage Volume of
`0.05 ml (1.25 mg). In one embodiment, the extractable vol
`ume from the Syringe (that is the amount of product obtain
`able from the Syringe following filling, taking into account
`loss due to dead space in the syringe and needle) is about 0.09
`ml.
`0024. In one embodiment the length of the syringe body is
`between about 45mmandabout 50 mm, the internal diameter
`is between about 4 mm and about 5 mm, the till volume is
`between about 0.12 and about 0.3 ml and the dosage volume
`is between about 0.03 ml and about 0.05 ml.
`0025. As the syringe contains a medicament solution, the
`outlet may be reversibly sealed to maintain sterility of the
`medicament. This sealing may beachieved through the use of
`a sealing device as is known in the art. For example the
`OVSTM system which is available from Vetter Pharma Inter
`national GmbH.
`0026. It is typical to siliconise the syringe in order to allow
`ease of use, i.e. to apply silicone oil to the inside of the barrel,
`which decreases the force required to move the stopper. How
`ever, for ophthalmic use, it is desirable to decrease the like
`lihood of silicone oil droplets being injected into the eye. With
`multiple injections, the amount of silicone droplets can build
`up in the eye, causing potential adverse effects, including
`
`Novartis Exhibit 2044.004
`Regeneron v. Novartis, IPR2020-01318
`
`

`

`US 2014/0012227 A1
`
`Jan. 9, 2014
`
`“floaters' and an increase in intra-ocular pressure. Further
`more, silicone oil can cause proteins to aggregate. A typical 1
`ml syringe comprises 100-800 ug silicone oil in the barrel,
`though a survey of manufacturers reported that 500-1000 ug
`was typically used in pre-filled syringes (Badkar et al. 2011,
`AAPS PharmaSciTech, 12(2):564-572). Thus, in one
`embodiment, a Syringe according to the invention comprises
`less than about 800 Lug (i.e. about less than about 500 ug, less
`than about 300 ug, less than about 200 ug, less than about 100
`ug, less than about 75 ug, less than about 50 ug, less than
`about 25 ug, less than about 15 Jug, less than about 10 ug)
`silicone oil in the barrel. If the syringe comprises a low level
`of silicone oil, this may be more than about 1 g, more than
`about 3 Jug, more than about 5 ug, more than about 7 Jug or
`more than about 10 ug silicone oil in the barrel. Thus, in one
`embodiment, the Syringe may comprise about 1 ug-about 500
`ug, about 3 ug-about 200 ug, about 5ug-about 10 ug or about
`10 ug-about 50 ug silicone oil in the barrel. Methods for
`measuring the amount of silicone oil 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. Various types of sili
`cone oil are available, but typically either DC360 (Dow Corn
`ing(R); with a viscosity of 1000 cB) or DC365 emulsion (Dow
`Corning R; DC360 oil with a viscosity of 350cP) are used for
`Syringe siliconisation. In one embodiment, the pre-filled
`syringe of the invention comprises DC365 emulsion.
`0027. During 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
`reducing the siliconisation levels far below the current stan
`dard to the levels discussed here. This is in contrast to con
`ventional thinking that would suggest that if you decrease the
`silicone oil level, the forces required would increase (see e.g.
`Schoenknecht. AAPS National Biotechnology Conference
`2007. Abstract no. NBC07-000488, which indicates that
`while 400 ug silicone oil is acceptable, usability improves
`when increased to 800 ug). 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 deliv
`ery may be made more difficult 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 100 ug-about 800 ug silicone oil.
`In one embodiment the glide/slide force for the stopper within
`the pro-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 or
`less 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 mea
`Sured at a stopper travelling speed of 190 mm/min. In one
`embodiment, the forces are measured with a 30Gx0.5 inch
`needle attached to the Syringe. In one embodiment, the
`Syringe has a nominal maximal fill Volume of between about
`0.5 ml and 1 ml, contains less than about 100 ug silicone oil
`and has a break loose force between about 2N to 5N.
`
`0028. In one embodiment the syringe barrel has an internal
`coating of silicone oil that has an average thickness of about
`450 nm or less (i.e. 400 nm or less, 350 nm or less, 300 nm or
`less, 200 nm or less, 10 nm or less, 50 nm or less, 20 nm or
`less). Methods to measure the thickness of silicone oil in a
`Syringe are known in the art and include the rap. ID Layer
`Explorer(R) Application, which can also be used to measure
`the mass of silicone oil inside a Syringe barrel.
`0029. 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
`and fill line. A further way to reduce silicone oil and inorganic
`silica levels in a pre-filled syringe is to avoid the use of
`silicone tubing in filling lines, for example between storage
`tanks and pumps.
`0030 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 >50 um in diameter per ml. In one embodi
`ment, the ophthalmic solution comprises no more than 5
`particles >25 um in diameter per ml. In one embodiment, the
`ophthalmic solution comprises no more than 50 particles >10
`um in diameter per ml. In one embodiment, the ophthalmic
`Solution comprises no more than 2 particles >50 um in diam
`eter per ml, no more than 5 particles >25um in diameter per
`ml and no more than 50 particles >10 Lum 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 sufficient for the syringe
`to meet USP789.
`
`VEGF Antagonists
`
`Antibody VEGF Antagonists
`
`0031 VEGF is a well-characterised signal protein which
`stimulates angiogenesis. Two antibody VEGF antagonists
`have been approved for human use, namely ranibizumab (Lu
`centis(R) and bevacizumab (AvastinR).
`
`Non-Antibody VEGF Antagonist
`
`0032. In one aspect of the invention, the non-antibody
`VEGFantagonist is an immunoadhesin. One Such immuoad
`hesin is aflibercept (EyleaR), which has recently been
`approved for human use and is also known as VEGF-trap
`(Holashet al. (2002) PNAS USA99: 11393-98: Riely & Miller
`(2007) Clin Cancer Res 13:4623-7s). Aflibercept is the pre
`ferred non-antibody VEGFantagonist for use with the inven
`tion. 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 con
`tains 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 K1 cells. Each monomer can
`have the following amino acid sequence (SEQID NO: 1):
`
`Novartis Exhibit 2044.005
`Regeneron v. Novartis, IPR2020-01318
`
`

`

`US 2014/0012227 A1
`
`Jan. 9, 2014
`
`SDTGRPFWEMYSEIPEIIHMTEGRELWIPCRWTSPNITWTLKKFPLDTLIPDGKRI IWDSRKGFIISNATY
`
`KEIGLLTCEATVNGHLYKTNYLTHROTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPS
`
`SKHOHKKLVNRDLKTOSGSEMKKFLSTLTIDGVTRSDOGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPP
`
`CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYWDGVEVHNAKTKPREEOYNST
`
`YRVVSVLTVLHODWLNGKEYKCKVSNKALPAPIEKTISKAKGOPREPOVYTLPPSRDELTKNOWSLTCLVK
`
`GFYPSDIAVEWESNGOPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWOOGNWFSCSVMHEALHNHYTOKSL
`
`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 mono
`CS.
`0033. Another non-antibody VEGF antagonist immu
`noadhesin currently in pre-clinical development is a recom
`binant 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
`VEGFR1/Flt-1; these domains are fused to a human IgGFc
`protein fragment (Li et al., 2011 Molecular Vision 17:797
`803). This antagonist binds to isoforms VEGF-A. VEGF-B
`and VEGF-C. The molecule is prepared using two different
`production processes resulting in different glycosylation pat
`terns 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 (SEQID NO:2):
`
`0035 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
`WO2O1 O/O6O748 and WO2O11/135067.
`0036 Further specific antibody mimetics with VEGF
`antagonist activity are the 40 kD pegylated anticalin PRS-050
`and the monobody angiocept (CT-322).
`0037. The afore-mentioned non-antibody VEGF antago
`nist may be modified to further improve their pharmacoki
`netic properties or bioavailability. For example, a non-anti
`body 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 present in the protein
`sequence of the natural protein from which the VEGFantago
`nist was derived.
`0038 Variants of the above-specified VEGF antagonists
`that have improved characteristics for the desired application
`
`MWSYWDTGWLLCALLSCLLLTGSSSGGRPFWEMYSEIPEIIHMTEGRELWIPCRWTSPNITWTLKKFPLDT
`
`LIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHROTNTIIDWVLSPSHGIELSVGEK
`
`LVLNCTARTELNVGIDFNWEYPSSKHOHKKLVNRDLKTOSGSEMKKFLSTLTIDGVTRSDOGLYTCAASSG
`
`LMTKKNSTFWRWHEKPFWAFGSGMESLWEATWGERWRLPAKYLGYPPPEIKWYKNGIPLESNHTIKAGHWL
`
`TIMEVSERDTGNYTVILTNPISKEKOSHVVSLVVYWPPGPGDKTHTCPLCPAPELLGGPSVFLFPPKPKDT
`
`LMISRTPEVTCVVVDVSHEDPEVKFNWYWDGVEVHNAKTKPREEOYNSTYRVVSWLTVLHODWLNGKEYKC
`
`KVSNKALPAPIEKTISKAKGOPREPOVYTLPPSRDELTKNOWSLTCLVKGFYPSDIAVEWESNGOPENNYK
`
`ATPPVLDSDGSFFLYSKLTVDKSRWOOGNWFSCSVMHEALHNHYTOKSLSLSPGK
`
`and, like VEGF-trap, can be present as a dimer. This fusion
`protein and related molecules are further characterized in
`EP1767546.
`0034. Other non-antibody VEGFantagonists include anti
`body mimetics (e.g. Affibody(R) molecules, affilins, affitins,
`anticalins, avimers, Kunitz domain peptides, and monobod
`ies) with VEGF antagonist activity. This includes recombi
`nant binding proteins comprising an ankyrin repeat domain
`that binds VEGF-A and prevents it from binding to VEGFR
`2. One example for such a molecule is DARPintR) MP01 12.
`The ankyrin binding domain may have the following amino
`acid sequence (SEQ ID NO:3):
`
`may be produced by the addition or deletion of amino acids.
`Ordinarily, these amino acid sequence variants will have an
`amino acid sequence having at least 60% amino acid
`sequence identity with the amino acid sequences of SEQID
`NO: 1, SEQID NO: 2 or SEQID NO:3, preferably at least
`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 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
`
`GSDLGKKLLEAARAGODDEVRILMANGADVNTADSTGWTPLHLAVPWGHLEIVEWLLKYGADVNAKDFQGW
`
`TPLHLAAAIGHQEIVEWLLKNGADVNAQDKFGKTAFDISIDNGNEDLAEILOKAA
`
`Novartis Exhibit 2044.006
`Regeneron v. Novartis, IPR2020-01318
`
`

`

`US 2014/0012227 A1
`
`Jan. 9, 2014
`
`ID NO: 1, SEQID NO: 2 or SEQID 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.
`0039. 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 FASTA, two polypep
`tides 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 a default gap penalty, and a scoring matrix Such as PAM
`250 a standard scoring matrix; see Dayhoffet al., in Atlas of
`Protein Sequence and Structure, Vol. 5, Supp. 3 (1978) can be
`used in conjunction 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 introduced into the
`longer sequences in order to align the two sequences.
`0040 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-antibody VEGF antagonist of the inven
`tion are preferably proteinaceous, but may include modifica
`tions that are non-proteinaceous (e.g., pegylation, glycosyla
`tion).
`Therapy
`0041. The syringe of the invention may be used to treat an
`ocular disease, including but not limited to choroidal neovas
`cularisation, age-related macular degeneration (both wet and
`dry forms), macular edema secondary to retinal vein occlu
`sion (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.
`0042. Thus the invention provides a method of treating a
`patient Suffering from of an ocular disease selected from
`choroidal neovascularisation, wet age-related macular degen
`eration, macular edema secondary to retinal vein occlusion
`(RVO) including both branch RVO (bRVO) and central RVO
`(cRVO), choroidal neovascularisation secondary to patho
`logic myopia (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 invention. This method pref
`erably 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.
`0043. In one embodiment, the invention provides a
`method of treating an ocular disease 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 patho
`logic myopia (PM), diabetic macular edema (DME), diabetic
`retinopathy, and proliferative retinopathy, comprising admin
`istering a non-antibody VEGF antagonist with a pre-filled
`Syringe of the invention, wherein the patient has previously
`received treatment with an antibody VEGFantagonist.
`
`Kits
`
`0044 Also provided are kits comprising the pre-filled
`Syringes of the invention. In one embodiment. 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 sterilisa
`tion, for example terminal sterilisation.
`0045. Such a kit may further comprise a needle for admin
`istration of the VEGFantagonist. If the VEGFantagonist is to
`be administered intravitreally, it is typical to use a 30-gaugex
`/2 inch needle, though 31-gauge and 32-gauge needles may
`be used. For intravitreal administration, 33-gauge or
`34-gauge needles could alternatively 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.
`
`Sterilisation
`
`0046. As noted above, a terminal sterilisation process may
`be used to sterilise the Syringe and Such a process may use a
`