DocCode - SEQ.TXT
`
`SCORE Placeholder Sheet for IFW Content
`
`Application Number: 13750352
`
`Document Date: 01/25/2013
`
`The presence of this 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
`database.
`
`• Sequence Listing
`
`Since this was an electronic submission, there is no physical artifact folder, no artifact folder is
`recorded in PALM, and no paper documents or physical media exist. The TIFF images in the
`IFW record were created from the original documents that are stored in SCORE.
`
`To access the documents in the SCORE database, refer to instructions developed by SIRA.
`
`At the time of document entry (noted above):
`• Examiners may access SCORE content via the eDAN interface.
`• Other USPTO employees can bookmark the current SCORE URL
`(http://es/ScoreAccessWeb/).
`• External customers may access SCORE content via the Public and Private PAIR
`interfaces.
`
`Form Revision Date: February 8, 2006
`
`Regeneron Exhibit 1002.0001
`
`

`

`··---·-······---
`
`~-----···
`
`·---·1-DocKet Nurr1ber I PAT055157-US-NP
`EU,l~G av "EXPRESS MAIL" UNDER 37 CFB.J.J.Q
`I
`L_= Express Mail Label N1.imbe-r - - - -
`
`··-----~~=--~~te of Deposit
`
`--···············,
`
`"']
`i
`
`____ J
`
`Commissioner for Patent&
`PO Box 1450
`Alexandria, VA 22313-·1450
`UTILITY PATENT APPLICATION TRANSMffT AL AND FEE SHEET
`Transmitted herewith for filing under 37 CFR §1.53(b) ls the utmty patent application of
`
`/\pp!icant (or identifier): Sigg, Juergen et a!.
`
`Title:
`
`SYRINGE
`
`Enclosed are:
`1. 0 Specification (Including Claims and Abstract) - 27 pages
`0 Drawings - ·1 sheets
`2..
`3. 0 Executed Declaration and Power of Attorney (original or copy)
`LJ Microfiche Computer Program (appendix)
`4.
`Nucleotide and/or Amino Acid Sequence Submission
`5.
`CJ Computer Readable Copy
`0 PaperCopy
`D Statement Verifying Identity of Above Copies
`Preliminary Amendment
`Assignment Papers (Cover Sheet & Document(s))
`English Translation of
`Information Disclosure Statement
`Certified Copy of Priority Document(s)
`Return Receipt Postcard
`Application Data Sheet
`Other: a) Submission of Sequence Listing Including Staternent of Verification ( ·1
`Sheet)
`b) Unexecuted Declaration for Utility Application using an Application Data
`Sheet (5 Sheets)
`c) Transmittal for Power of Attorney to one or more registered Practitioners
`and Power of Attorney by Applicant (2 s!1eets)
`
`6. D
`7. D
`[]
`8.
`tSl
`9,
`10. D
`11. 0
`12. 0
`13. 0
`
`Filing fee calculation:
`D Before calculating the filing fee, please enter the enclosed Preliminary Amendment.
`D Before calculating the filing fee, please cancel claims
`
`Regeneron Exhibit 1002.0002
`
`

`

`fa-as-ic-Fi-!in_g_F_e_e__________________________
`r Search Fee
`L $ _ 62Q __
`-·········-······-·-·-···--·---
`r Examination.f.~-~ - - - - - - - - -
`250
`. $
`·-··-······-----
`·--------------------··-··-··-----------$ _ _ _ 0
`0
`$
`
`.·--····-=--~~~----~rl. ___ 3_8_0.
`
`[_-=R-at~----.·_·------+---···-..]
`Number I
`,,, I$
`8 _:__ __ ---- 250 ·· ··-·=r $---0-,
`l X
`
`}
`~Multiple Dependent Claim Fee($
`}
`: Foreign Lan~uage Surc_~_?..iByJ .(~--
`i
`Number I
`For
`I
`i
`l______
`______ ,
`Filed
`Extra
`!
`i -20
`Extra
`12
`32
`1 Tota! Claims
`Claims
`-··--------- -----
`!
`...
`independent
`0
`Claims
`'
`-------
`-•-•n-•••---·· ••--••._
`~22!!_cation Size Fee,-------,--~-------·---···········[::]
`Extra ~umber of each additbnal
`Rate
`Total
`50 or fraction thereof
`Sheets
`Sheets
`-.-----.'----+----+-
`~nd~d uo to a whole numb~.
`i $
`0
`0
`- 100
`28
`X
`
`1
`
`. ,
`1
`
`I
`
`··,:)
`
`i
`I X
`i
`'
`
`60
`
`120
`
`[---·-------------------------~---·---······-··-- TOTA~ F!UNG FEE [ $ -·----~!~~::!
`
`!--··-·········
`: i $
`0
`
`310
`
`~ Please charge Deposit Account No. ·, S-0134 in the name of Novartis in the amount of
`$1970 . .An additional copy of this paper is endosed. 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 crecm any overpayment, to Deposit
`Account No. 19-0134 in the name of Novartis.
`Please address au correspondence to the address associated with Customer No. 00'1095,
`which is currently:
`
`Novartis Pharmaceuticals Corporation
`One Health Plaza, Bldg. 101
`East Hanover, NJ 07936
`
`Please direct all telephone ca!!s to the undersigned at the number given below, and all
`telefaxes to +1 9737818265.
`
`Respectfully submitted,
`
`/ Andrew Holmes/
`--------·---··-·-·····---··-·········
`Andrew Holmes
`Agent for Applicant
`Reg. No. 5·1 ,813
`
`Novartis Pharmaceuticals Corporation
`One Health Plaza, Bldg. 101
`East Hanover, NJ 07936
`+1 8627785816
`
`Date: ,January 25, 2013
`
`2
`
`Regeneron Exhibit 1002.0003
`
`

`

`TECHNICAL FIELD
`
`SYRIN(;E
`
`The present invention relates to a syringe, particularly to a small volume syringe such as a
`syringe suitable for ophthalmic injections.
`
`5 BACKGROUND ART
`
`IO
`
`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,
`It is important for patient safety and medicament
`or a caregiver, to inject the 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 or a sterilising gas.
`
`For small volume syringes, 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 compromise sterility. Incorrect handling of the syringe can also
`
`pose risks to product sterility.
`
`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 of sterilisation 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
`
`There is therefore a need for a new syringe construct which provides a robust seal for its content,
`but which maintains ease of use.
`
`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
`
`30
`
`-1-
`
`Regeneron Exhibit 1002.0004
`
`

`

`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 bet\veen 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
`
`5
`
`chamber, characterised in that the fluid comprises an ophthahnic solution. 1n one embodiment.
`
`the ophthalmic solution comprises a VEGF-antagonist.
`
`1n one embodiment, the syringe is suitable for ophthaln1ic injections, rrwre particularly
`
`intravitrea1 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 1ubricant
`
`1 O
`
`In one embodiment, despite the lo\</ silicone oil level, the stopper break loose and slide force is
`
`less than 20N.
`
`For ophthalmic injections, it is particularly impmi:ant fix the ophthalmic solution to have
`particularly low particle content. rn one embodiment, the syringe meets US Pharmacopeia
`standard 789 (USP789).
`
`l5
`
`Syringe
`
`The body of the syringe may be a substantially cylindrical sheH, or may indude a substantially
`
`cylindrical bore with a non circular outer shape. The outlet end of the body includes an outlet
`
`through which a t1uid housed vdthin the variable volume chamber can be expelled as the volume
`
`of said chamber is reduced. The outlet may comprise a projection frorn the outlet end through
`
`20 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
`
`25 made directly between the syringe and accessory, or via the sealing device. The body extends
`
`along a first axis fron1 the outlet end to a rear end.
`
`The body may be made from a plastic rnaterial (e.g. a cydic 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 co1nprise a priming mark. This allows the physician to align a pre~determined part of
`
`30
`
`the stopper (such as the tip of the front surface or one of the drcumferentia! ribs, discussed later)
`
`or plunger with the mark, thus expelling excess ophthalmic solution and any air bubbles from the
`
`-2-
`
`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 from rubber, silicone or other suitable resiliently deformable material.
`
`The stopper may be substantially cylindrical and the stopper may include one or n1ore
`
`5
`
`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 ,:vith an internal surface of the
`
`syringe body, T11e 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
`
`10
`
`plunger to the stopper using a snap fit feature or thread connection in a knmvn manner. The
`
`stopper may be substantially rotationally symmetric about an axis through the stopper.
`
`0l11e 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 nortion mav
`'
`comprise a substantially disc shaped poition, the radius of the disc extending substantially
`
`15
`
`perpendicular to the axis along which the rod extends. The user (:ontact pmtion could be any
`
`suitable shape. TI1e 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 ofthe body. The backstop may be
`
`20
`
`removable from the syringe. ff the syringe body includes tem1inal flanges at the end opposite the
`
`outlet end the backstop may be configured to substantiaily samhvich terminal flanges of the body
`
`as this prevent movement of the backstop in a direction paral1el to the first axis.
`
`The rod may comprise at least one rod shoulder directed away from the outlet end and the
`
`backstop rnay include a backstop shoulder directed towards the outkt end to cooperate ·with the
`
`25
`
`rod shoulder to substantially prevent movernent of the rod away from the outlet end when the
`
`backstop shoulder and rod shoulder are in contact. Restriction of the rnovement of the rod away
`
`frorn the outlet end can help to maintain sterility during terminal sterilisation operations, or other
`
`operations in which the pressure within the variable vohim(~ .:.:hamber 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 cau::se 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
`
`.. 3 ..
`
`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 vvithin 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 exampie a circumferential rib, closest to the outlet and a seai of the stopper,
`
`5
`
`for example a circumferential rib, farthest from the outlet The distance between these two seals
`
`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
`
`1 O
`
`separated in a direction along the first axis by at !east 3mm, by at ieast 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, In one embodiment there are a total of three circumferential ribs.
`
`A stopper with such an enhanced sterility zone can also provide protection for the injectable
`
`15 medicarnent during a terminal sterilisation process.
`
`f\fore 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. Hmvever, increasing the number of ribs can increase the friction between
`
`the stopper and syringe body, reducing ease of use. \\lhile this may· be oven.:ome by increasing
`
`the siliconisation of the syringe, such an increase in silicone oil levels is particularly undesirable
`
`20
`
`for syringes for ophthalmic use.
`
`The rod shoulder may be arranged within the external dbm.eter of the rod, or may be arranged
`
`outside the extema! diameter of the rod, By providing a shoulder that extends beyond the
`externa! 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
`
`25
`
`axis. The rod shoulder may co1nprise any suitable shoulder forming elements on the rod, but in
`
`one embodiment the rod shoulder comprises a substantially disc shaped portion on the rod.
`
`1n one embodiment of the syringe, when arranged \i.1-\th the plunger contact surfi:Ke in contact
`
`with the stopper and the variable volurne chamber is at its intended maximum volume there is a
`
`clearance of no m.ore than about 2mm between the rod shoulder and backstop shoulder. ln some
`
`30
`
`embodiments there is a clearance of less than about ] . .5 mm and in some less than about ] mm.
`
`This distance is selected to substantially limit or prevent excessive rearward (away from the
`
`outlet end) movement of the stopper.
`
`A-
`
`Regeneron Exhibit 1002.0007
`
`

`

`In one embodiment the variable volume chamber has an internal diameter greater than 5mm or
`
`6mm, or less than 31mn or 4mm. The internal diameter may be between 3mm and 6mm, or
`
`between 4mm and 5rnm.
`
`1n another embodiment the syringe is dirnensioned so as to have a nominal rnaximum fiU volume
`
`5
`
`of bet\:veen about 0.1ml and about L5rnL In certain embodirnents the nominal maximum fill
`
`volume is between about 0.5ml and about 1ml.
`
`ln certain embodirnents the nominal maximum
`
`fill volume is about 0.5ml or a.bout 1ml, or a.bout L5ml.
`
`The length of the body of the syringe may be Jess than 70mm, fess than 60mrn or less than
`
`50mm. In one embodiment the length of the syringe body is between 45mm and 50mm.
`
`IO
`
`In one embodiment the syringe is filled with between about 0.01ml and about 1.5ml (for
`
`example between about 0.05ml and about "lml, between about 0.1ml and about 0.5ml, between
`
`about 0.15ml and about 0.175ml) of a VEGF antagonist solution. In ont:'. embodiment, the
`
`syringe is filled \vith 0. 165ml of a VEGF antagonist solution, Of course, typically a syringe is
`
`filled with more than the desired dose to be administered to the patient, to take into account
`
`15
`
`wastage due to "dead space" within the syringe and needle. There may also be a certain amount
`
`of wastage vvhen the syringe is primed by the physician, so that it is ready to inject the patient.
`
`Thus, in one embodiment, the syringe is filled with a dosage volume (Le. the volume of
`
`medicament intended for delivery to the patent) of between about 0.01ml and about 1.5ml (e.g.
`
`behveen about 0.05ml and about 1ml, behveen about 0,1ml and about 0.5ml) of a VEGF
`
`20
`
`antagonist solution. In one embodiment, the dosage volume is between about 0.03ml and about
`
`0.05ml. For example, for Lucentis, the dosage volurne is 0.05m.! or 0.03ml (0.5mg or 0.3mg) of a
`
`l Omg/m.1 injectable medicament solution; for Eylea, the dosage vo.lume is 0.05ml of a 40mg/ml
`
`injectable medicament solution, Although unapproved for ophthairnk indications, bevacizumab
`
`is used off-label in such ophthalmic indications at a concentration of 251ng/ml; typica Hy at a
`
`25
`
`dosage volume of 0.05ml ( 1.25mg). ln one embodiment, the extractable volume from the syringe
`
`(that is the amount of product obtainable frorn the syringe following filling, taking into account
`
`loss due to dead space in the syringe and needle) is about 0.09ml.
`
`1n 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
`
`30
`
`and about 0.3ml and the dosage volume is between about 0.03ml and about 0.05ml.
`
`-5-
`
`Regeneron Exhibit 1002.0008
`
`

`

`As the syringe contains a medkmnent solution, the outlet may be reversibly sealed to maintain
`
`sterility of the medicament. This sealing 1nay be achieved through the use of a sealing device as
`is known in the art for example the OVS1
`
`M system which is available from Vetter Pharma
`
`International GmbH.
`
`5
`
`It is typical to siliconise the syringe in order to aHow ease of use, i.e. to apply silicone oil to the
`
`inside of the barrel, which decreases the force required to move tbe swpper. However, fbr
`
`ophthalmic use, it is desirable to decrease the likelihood of sfficone 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 "floaters" and an increase in intra-ocular pressure,
`
`10
`
`Furthermore, silicone oil can cause proteins to aggregate. A typical 1ml syringe comprises 100-
`
`SOOµg silicone oil in the barrel, though a survey of manufacturers reported that 500-l OOOµg 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 cornprises less than about 800µg
`
`(Le. about less than about 500~tg, less than about :.rno~ig, less than about 200~tg, less than about
`
`15
`
`100µg, less than about 75µg, less than about 50µg, less than about 25~tg, less than about l5iig.
`
`less than about lOµg) silicone oil in the barrel. If the syringe comprises a low kvel of silicone
`
`oil, this may be more than about lµg, more than about 3~tg, more than about 5~tg, more than
`
`about 7µg or more than about 10pg silicone oil in the barreL Thus, in one embodiment, the
`
`syringe may comprise about l µg-about 500Fg, about 3µg-about. 200µg, about 5~tg-about 1 OO~tg
`
`20
`
`or about 1 O~tg-about SOµg sllicone 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 tht~ oil diluted in a suitable
`
`solvent. Various types of silicone oil are available, but typically either DC360 (Dow Corning@';
`
`with a viscosity of 1000cP) or DC365 emulsion (Dow Coming"1'; DC:360 oil with a viscosity of
`
`25
`
`350cP) are used for syringe silkonisation, In one embodinwnt, the pre-filled syringe of the
`
`invention comprises DC365 emulsion.
`
`During testing it ;,vas surpdsingly found that, for syringes having small dimensions, such as those
`
`discussed above, and particularly those described in cor\junction \Vith the Flgures below, the
`
`break loose and sliding forces for the stopper within the syringe are substantially unaffected by
`
`30
`
`reducing the silkonisation Ieve1s far below the cunent standard to the levels discussed here. This
`
`is in contrast to conventional thinking that would suggest that if you decrease the silicone oil
`
`level, the forces required would increase (see e.g. Schoenlrnecht AAPS National Biotechnology
`
`Conference 2007 - Abstract no. NBC07~000488, which indicates that while 400pg silicone oil is
`
`-6-
`
`Regeneron Exhibit 1002.0009
`
`

`

`acceptable, usability improves when increased to 800µg). 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 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
`1 OOµg-about 800µg silicone oil. In one embodiment the glide/slide force for the stopper within
`the pre-filled syringe is less than about 1 lN 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 l lN or less
`than 9N, less than 7N, less than 5N or between about 2N to SN. 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 maximal fill
`volume of between about 0.5ml and 1ml, contains less than about lOOµg silicone oil and has a
`break loose force between about 2N to 5N.
`
`In one embodiment the syringe barrel has an internal coating of silicone oil that has an average
`thickness of about 450nm or less (i.e. 400nm or less, 350nm or less, 300nm or less, 200nm or
`less, 100nm or less, 50nm or less, 20nm 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® Application, which can
`also be used to measure the mass of silicone 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 and fill line. A further way to reduce silicone oil and inorganic silica levels in a pre(cid:173)
`filled syringe is to avoid the use of silicone 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 ~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
`more than 50 particles ~lOµm in diameter per ml. In one embodiment, the ophthalmic solution
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`-7-
`
`Regeneron Exhibit 1002.001 O
`
`

`

`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,
`
`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.
`
`VEGF Antagonists
`
`Antibody VEGF antagonists
`
`VEGF is a well-characterised signal protein which stimulates angiogenesis. Two antibody VEGF
`
`antagonists have been approved for human use, namely ranibizumab (Lucentis®) and
`
`10
`
`bevacizumab (A vastin®).
`
`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 Fe portion of human IgG 1. 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 Kl cells. Each monomer can have the
`
`following amino acid sequence (SEQ ID NO: 1):
`
`SDTGRPFVEMYSEIPEIIHM:TEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATY
`
`KEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPS
`
`25
`
`SKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPP
`
`CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
`
`YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK
`
`GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL
`
`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.
`
`-8-
`
`Regeneron Exhibit 1002.0011
`
`

`

`Another non-antibody VEGF antagonist inummoadhesi11 currently in pre-clinical development is
`
`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
`VEGFRl/Flt-1; these domains are fused to a human IgG Fe prottin fragment (Li et al., 2011
`
`5 Molecular Vision 17:797-803). This antagonist binds to isoforrns VEGF-A, VEG11-B and VEGF-·
`
`C. The molecule is prepared using two different production processes resulting ln different
`
`glycosylation patterns on the final proteins. The two glycoforms axe referred to as KH902
`
`(conbercept) and KB906. The fusion protein can have the follmving amino add sequence (SEQ
`
`ID N0:2):
`
`lO
`
`15
`
`l\'lVSY\rJD'I'GVLLCALLSCLLL'l'GSSSGGRPFVEMYSEIPEIIHMTEGRELVIPCR\J"TSPNITVTLKKFPLDT
`
`LIPDGKRII\'v-:DSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYL'l'HRQTNTJ.IDVVL,SPSHGIELSVGEK
`
`Li/LNCTARTELNVGIDF'rNJEYPSSKHQHKKLVNRDLK'l\.~SGSEi)1.KKFLST'L'rIDGVTP.SDQGLYTCA.,\SSG
`
`LNTKIZNS'l'FVRVHEKPFVAFGSGMESLVEATVGERVRLPAKYLGYPPPEIK\rvYK.!.\fGIPLESNHTIK.AGHVL
`
`TIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVYVPPGPGDK'l'H'.l'CPLCPAPELLGGPSVFLF'PPKPKDT
`
`LM1SR~'PEVrCV\/VDVS.HEDPEVKFNWYVDGVEV1iNAKTKPREEQYNS'I'YR\TVSVLTVLHQm,JLNGKEYKC
`
`KVSNKALPAPIEKTISKAKGQPREPQ\T'JTLPPSRDELTK.NQVSLTCINKGFYPSDIAVEWESNGQPENNYK
`
`ATPPVLDSDGSF'.F'LYSKL'l'VDKSRWQQGNVFSCSV.ViHEALHffHY'l'QKSLSLSPGK
`
`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 mimetks (e.g. Affibody@ molecules,
`
`affilins, affitins, anticatins, avimers, Kunitz domain peptides, and monobodies) with VEGF
`
`antagonist activity. This includes recombinant binding proteins co.rnprising an ankyrin repeat
`
`domain that binds VEGF-A and prevents it from binding to VEGFR-2. One example for such a
`
`rnokcule is DAR.Pin® MPOl 12. The ankyrin binding domain may have the following amino
`
`25
`
`acid sequence (SEQ ID NO: 3):
`
`GSDLGKKLLEAARAGQDDEVRILH.Al.~GADv1'JTADSTG\'\iTPLBLAVP\rvGHLEIVE'VLLKYGADVNAKDF'QGW
`
`'rPLHLAAAI GHQE IVEVLLKNGADVNAQDI<.FG.K'l'AFDI SI DNGNEDLAEI LQKAA
`
`Recombinant binding proteins comprising an ankyrin repeat domain that binds VEGF-A and
`
`prevents it from binding to VEGFRY2 are described in more detail in W020l0/060748 and
`
`30 W02011/135067.
`
`Further specific antibody mimetks vvith VEGF antagonist activity are the 4-0 kD pegylated
`
`anticalin PRS-050 and the monobody angiocept (CT-322)-
`
`-9-
`
`Regeneron Exhibit 1002.0012
`
`

`

`The <1fore-mentioned non-antlbody VEGF antagonist may be modified to further improve their
`
`pharmacokinetic propt~rt.ies 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 rnay be modified by glycosylation or the addition of further glycosy1ation sites not
`
`5
`
`present in the protein sequence of the natu.rai protein from which the VEGF antagonist was
`
`derived.
`
`Variants of tht, above-specified VEGF antagonists that have irnproved characteristics for the
`
`desired application may be produced by the addition or deletion of amino acids. Ordinarily, these
`arnino acid sequence variants wiU have an amino acid sequence having at least 6m10 amino acid
`sequence identity with the amino acid sequences of SEQ lD NO: l, SEQ m NO: 2 or SEQ lD
`NO: \ preferably at least 8(Y}o, more preferably at least 85%, more preferably at least 90%, and
`
`10
`
`most preferably at least 95°/o, including few example, 80%,, 81 %, 82%, 8.3%, 84%, 85?1o, 86%,
`
`87%, 88%i, 89%, 90%, 91 %, 92%, 93%>, 94%, 95%, 96%1, 97o/o, 98%,, 99%, and 100%. Identity
`
`or homology with respec:t to this sequence is defined herein as the percentage of amino acid
`
`15
`
`residues in the candidate sequence that are identical with SEQ ID NO: !, SEQ JD NO: 2 or SEQ
`
`lD NO: 3, after aligning the sequences and introducing gaps, if necessary, to achieve the
`
`maximum percent sequence identity, and not considering any conse:rvative substitutions as part
`
`of the sequence identity.
`
`Sequence identity can be determined by standard methods that are commonly used to compare
`
`20
`
`the similarity in position of the amino acids of two po1ypeptides. 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 sequenees or along a pre·
`
`determined portion of one OJ' 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
`Dayhoff et at, in Atlas of Protein Sequence and Structure, voL 5, supp. 3 (1978)] can be used in
`
`25
`
`conjunction with the computer program.
`
`For example, the percent kk,ntity 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 vvithin 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-
`
`-10--
`
`Regeneron Exhibit 1002.0013
`
`

`

`antibod:y VEGF antagonist of the invention are preferably proteinaceous, but n:rny include
`
`rnodifications that are non--proteinaceous (e.g., pegylation, glycosylation).
`
`Thempy
`
`The