I IIIII IIIIIIII Ill lllll lllll lllll lllll lllll lllll lllll lllll 111111111111111111
`US009220631B2
`
`c12) United States Patent
`Sigg et al.
`
`(IO) Patent No.:
`(45) Date of Patent:
`
`US 9,220,631 B2
`Dec. 29, 2015
`
`(54) SYRINGE
`
`(71) Applicant: Novartis AG, Basel (CH)
`
`(72)
`
`Inventors: Juergen Sigg, Loerrach (DE);
`Christophe Royer, Munich (DE);
`Andrew Mark Bryant, Basel Land
`(CH); Heinrich Martin Buettgen,
`Rheinfelden (CH); Marie Picci,
`Ranspack-le-bas (FR)
`
`(73) Assignee: Novartis AG, Basel (CH)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term ofthis
`patent is extended or adjusted under 35
`U.S.C. 154(b) by O days.
`
`(21) Appl. No.: 13/750,352
`
`(22) Filed:
`
`Jan.25,2013
`
`(65)
`
`Prior Publication Data
`
`US 2014/0012227 Al
`
`Jan. 9, 2014
`
`(30)
`
`Foreign Application Priority Data
`
`Jul. 30, 2012
`Oct. 23, 2012
`Nov. 16, 2012
`Nov. 16, 2012
`Nov. 16, 2012
`Nov. 23, 2012
`Nov. 23, 2012
`Dec. 3, 2012
`Jan. 23, 2013
`Jan. 23, 2013
`Jan. 23, 2013
`
`(EP) ..................................... 12174860
`(EP) ..................................... 12189649
`(AU) ................................ 2012101677
`(AU) ................................ 2012101678
`(DE) ..................... 202012011016U
`(DE) ..................... 20 2012 011 259 U
`(DE) ..................... 20 2012 011 260 U
`(EP) ..................................... 12195360
`(AU) ................................ 2013100070
`(AU) ................................ 2013100071
`(DE) ..................... 20 2013 000 688 U
`
`(51)
`
`Int. Cl.
`A61M5/00
`A61F 9/00
`A61M5/178
`
`(2006.01)
`(2006.01)
`(2006.01)
`
`(Continued)
`
`(52) U.S. Cl.
`CPC ............. A61F 9/0008 (2013.01 ); A61K 9/0019
`(2013.01); A61K 9/0048 (2013.01); A61K
`38/179 (2013.01); A61M 5/002 (2013.01);
`A61M 5/178 (2013.01); A61M 5/28 (2013.01);
`A61M 5/31 (2013.01); A61M 5/315 (2013.01);
`A61M 5/31505 (2013.01); A61M 5/31513
`(2013.01);
`
`(Continued)
`
`(58) Field of Classification Search
`CPC ........ A61K 9/0048; A61F 9/008; A61M 5/31
`USPC .................................................. 604/218, 294
`See application file for complete search history.
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`6,090,081 A * 7/2000 Sudo et al. .................... 604/230
`7,141,042 B2 * 11/2006 Lubrecht ....................... 604/230
`(Continued)
`
`FOREIGN PATENT DOCUMENTS
`
`AU
`AU
`
`12/2012
`2012101677 A4
`12/2012
`2012101678 A4
`(Continued)
`
`OTHER PUBLICATIONS
`
`Badkar et al., "Development of biotechnology products in pre-filled
`syringes:
`technical considerations and approaches", AAPS
`PharmaSciTech, vol. 12, No. 2, pp. 564-572, (Jun. 2011).
`(Continued)
`
`Primary Examiner - Aarti B Berdichevsky
`(74) Attorney, Agent, or Firm - Michael Mazza
`
`(57)
`
`ABSTRACT
`The present invention relates to a syringe, particularly to a
`small volume syringe such as a syringe suitable for oph(cid:173)
`thalmic injections.
`
`26 Claims, 1 Drawing Sheet
`
`Regeneron Exhibit 1001.001
`
`

`

`US 9,220,631 B2
`Page 2
`
`(51)
`
`Int. Cl.
`A61M5/315
`A61K9/00
`A61K 38/17
`A61M5/28
`A61M 5/31
`(52) U.S. Cl.
`CPC .................. A61M 2005/3104 (2013.01); A61M
`2005/3139 (2013.01)
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`7,303,748 B2 * 12/2007 Wiegand et al. ........... 424/134.1
`2006/0172944 Al *
`8/2006 Wiegand et al.
`514/12
`2006/0293270 Al
`12/2006 Adamis et al.
`2007/0190058 Al*
`8/2007 Shams ....................... 424/145.1
`2008/0312607 Al* 12/2008 Delmotte et al.
`604/230
`2010/0310309 Al
`12/2010 Abendroth et al.
`2011/0257601 Al
`10/2011 Furfine et al.
`2011/0276005 Al* 11/2011 Hioki et al.
`2012/0078224 Al
`3/2012 Lerner et al.
`2013/0012918 Al
`1/2013 Foster
`2014/0249484 Al *
`9/2014 Jones et al.
`
`604/187
`
`604/230
`
`FOREIGN PATENT DOCUMENTS
`
`CN
`DE
`EP
`EP
`EP
`JP
`JP
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`
`201578690 U
`10 2008 005938 Al
`0264273 A2
`0879611 A2
`2371406
`2001-104480
`2002241264 A2
`97/44068 Al
`2006047325 Al
`2006128564 Al
`WO 2007/035621 Al
`2007084765 A2
`WO 2007/149334 A2
`W02010/060748
`2010136492
`WO 2011/123722 Al
`W02011/135067
`WO 2012/134528 Al
`WO 2012/149040 A2
`2014/005728 Al
`
`9/2010
`7/2009
`4/1988
`11/1998
`10/2011
`4/2001
`8/2002
`11/1997
`5/2006
`12/2006
`3/2007
`7/2007
`12/2007
`6/2010
`12/2010
`10/2011
`11/2011
`10/2012
`11/2012
`1/2014
`
`OTHER PUBLICATIONS
`
`Ausubel et al., "Current Protocols in Molecular Biology", 7.7.18 of
`Current protocols in Molecular Biology, eds., supplement 30, ( 1987).
`Badkar et al., Analysis of Two Commercially Available Bortezomib
`Products Differences in Assay of Active Agent and Impurity
`Profile>> AAPS PharrnaSciTech, vol. 12, No. 2, pp. 564-572, (Jun.
`2011).
`
`Schoenknecht, "Requirements on pre-fillable glass suringes", AAPS
`National Biotechnology Conference 2007-Abstract No. NBC07-
`000488, 2007.
`Holash et al., "VEGF-Trap: A VEGF blocker with potent anitumor
`effects", PNAS USA, vol. 99, No. 17, pp. 11393-11398, (Aug. 20,
`2002).
`Riely & Miller, "Vascular Endothelial Growth Factor Trap in Non(cid:173)
`Small Cell lung Cancer", Clin Cancer Res, 13:4623-7s, (Aug. 1,
`2007).
`Li et al., "KH906, a recombinant human VEGF receptor fusion
`protein,
`is a new effective
`topical
`treatment
`for corneal
`neovascularization", Molecular Vision, 17:797-803,(Mar. 25, 2011).
`Smith & Waterman, "Comparison of Biosequences", Adv Appl.
`Math, 2:482-489, (1981).
`Chan et al: "Syringe Siliconization Process Investigation and Opti(cid:173)
`mization" Journal of Pharmaceutical Science and Technology, Issue
`66, pp. 137, 147-148, Mar. 2012.
`Lankers: "The Relationship Between Silicone Layer Thickness, Free
`Silicone Oil and Protein Aggregation in Prefilled Syringes" 2010
`AAPS National Biotechnology Conference San Francisco, Slides 25,
`39, 46, May 19, 2010.
`Majumdar et al: "Evaluation of the Effect of Syringe Surfaces on
`Protein Formulations" Journal of Pharmaceutical Sciences, Issue
`100, pp. 2563-2573, Jul. 2011.
`Bakri and Ekdawi: "Intravitreal Silicone Oil Droplets after
`Intravitreal Drug Injections" Retina, Issue 28, pp. 996-1001, Jul.
`2008.
`Daikyo Ru Crystal Zenith Insert Needle Syringe System, West Deliv(cid:173)
`ering Innovative Solutions, 2010.
`Injection
`Intravitreal
`a Safe
`Meyer
`et
`al:
`"Steps
`for
`Technique",Meyer et al. "Steps for a Safe Intravitreal Injection
`Technique"Retinal Physician, p. 3, Jul. 1, 2009.
`"Biopharmaceuticals-SPE applications", RapID Particle Systems,
`Single Particle Explore, D6a, Sep. 28, 2015, http //www.particle(cid:173)
`explorer.com/yourapplications/biopharaceuticals/index.htrnl[Sep.
`16, 2015 11:12:45].
`Email dated Sep. 9, 2015 from Elizabeth Scuderi, Senior meeting
`Manager, AAPS to Teresa Hornnch re Inquiry about publication of
`conference abstract.
`Tibor Hlobik: "Reducing quality risks to drug products and meeting
`needs of patients with enhanced components for prefilled syringe
`systems". West Delivering
`Innovative
`Solutions, www.
`ondrugdelivery.com, 2012 No. 33, pp. 32-34.
`Summary of Product Characteristics-Zaltrap (undated).
`"Ranibizumab", Scientific Discussion, EMEA, 2007, pp. 1-54.
`"Avastin", Scientific Discussion, EMEA, 2005, pp. 1-61.
`Melmet Selim Kocabora, et al. "Intreavitreal silicone oil droplets
`following pegaptanib injection", Acta Ophthalmologica, 2010 e44-
`345.
`N. Clunas, et al: "Ranibizumab pre-filled syringe: recently approved
`innovation in the Eurpean Union with the potential to reduce infec(cid:173)
`tion risk, improve does accuracy, and enhance efficient treatment
`administration". Congress on Controversies in Ophthamology,
`Abstract, 2014.
`"COPHy Poster List-Group A"(Poster 17), The 5th World congress
`on Controversies in Opthalmology (COPHy) Mar. 20-23, 2014,
`Lisbon, Portugal.
`
`* cited by examiner
`
`Regeneron Exhibit 1001.002
`
`

`

`U.S. Patent
`
`Dec. 29, 2015
`
`US 9,220,631 B2
`
`z
`I
`
`,-
`
`"
`
`t(
`
`">
`L
`
`"
`
`Fig 1
`
`r--. ...
`
`~ ' ~
`
`6
`
`/
`
`4-
`
`\L 3.&
`
`'30
`
`4-2.
`
`/6
`
`10
`
`I
`
`66
`
`Fig4
`
`Fig 5
`
`~
`6z..
`
`60
`
`Regeneron Exhibit 1001.003
`
`

`

`US 9,220,631 B2
`
`1
`SYRINGE
`
`TECHNICAL FIELD
`
`The present invention relates to a syringe, particularly to a
`small volume syringe such as a syringe suitable for oph(cid:173)
`thalmic injections.
`
`BACKGROUND ART
`
`15
`
`2
`For ophthalmic injections, it is particularly important for
`the ophthalmic solution to have particularly low particle con(cid:173)
`tent. In one embodiment, the syringe meets US Pharmacopeia
`standard 789 (USP789).
`5 Syringe
`The body of the syringe may be a substantially cylindrical
`shell, or may include a substantially cylindrical bore with a
`non circular outer shape. The outlet end of the body includes
`an outlet through which a fluid housed within the variable
`10 volume chamber can be expelled as the volume of said cham(cid:173)
`ber is reduced. The outlet may comprise a projection from the
`outlet end through which extends a channel having a smaller
`diameter than that of the variable volume chamber. The outlet
`may be adapted, for example via a luer lock type connection,
`for connection to a 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.
`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 embodi(cid:173)
`ment 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
`30 excess ophthalmic solution and any air bubbles from the
`syringe. The priming process ensures that an exact, pre-de(cid:173)
`termined 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
`more circumferential ribs around an outer surface of the stop-
`per, 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
`40 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
`45 stopper may be substantially rotationally symmetric about an
`axis through the stopper.
`The plunger comprises a plunger contact surface and
`extending from that a rod extends from the plunger contact
`surface to a rear portion. The rear portion may include a user
`50 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-
`55 able shape. The axis along which the rod extends may be the
`first axis, or may be substantially parallel with the first axis.
`The syringe may include a backstop arranged at a rear
`portion of the body. The backstop may be removable from the
`syringe. If the syringe body includes terminal flanges at the
`60 end opposite the outlet end the backstop may be configured to
`substantially sandwich terminal flanges of the body as this
`prevent movement of the backstop in a direction parallel to
`the first axis.
`The rod may comprise at least one rod shoulder directed
`away from the outlet end and the backstop may include a
`backstop shoulder directed towards the outlet end to cooper(cid:173)
`ate with the rod shoulder to substantially prevent movement
`
`Many medicaments are delivered to a patient in a syringe
`from which the user can dispense the medicament. If medi(cid:173)
`cament 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 steri- 20
`lised using heat or a sterilising gas.
`For small volume syringes, for example those for injec(cid:173)
`tions 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 diffi(cid:173)
`culties that are not necessarily associated with larger 25
`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(cid:173)
`promise sterility. Incorrect handling of the syringe can also
`pose risks to product sterility.
`Furthermore, certain therapeutics such as biologic mol(cid:173)
`ecules are particularly sensitive to sterilisation, be it cold gas
`sterilisation, thermal sterilisation, or irradiation. Thus, a care(cid:173)
`ful balancing act is 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(cid:173)
`cament must not be too high.
`There is therefore a need for a new syringe construct which
`provides a robust seal for its content, but which maintains
`ease of use.
`
`35
`
`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 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(cid:173)
`prises an ophthalmic solution. In one embodiment, the oph(cid:173)
`thalmic solution comprises a VEGF-antagonist.
`In one embodiment, the syringe is suitable for ophthalmic
`injections, more particularly intravitreal injections, and as
`such has a suitably small volume. The syringe may also be
`silicone oil free, or substantially silicone oil free, or may
`comprise a low level of silicone oil as lubricant. In one 65
`embodiment, despite the low silicone oil level, the stopper
`break loose and slide force is less than 20N.
`
`Regeneron Exhibit 1001.004
`
`

`

`US 9,220,631 B2
`
`3
`of the rod away from the outlet end when the backstop shoul(cid:173)
`der and rod shoulder are in contact. Restriction of the move(cid:173)
`ment 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 5
`chamber or outside the chamber may change. During such
`operations any gas trapped within the variable volume cham(cid:173)
`ber, 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 breach- 10
`ing of a sterility zone created by the stopper. This is particu(cid:173)
`larly important for low volume syringes where there are much
`lower tolerances in the component sizes and less flexibility in
`the stopper. The term sterility zone as used herein is used to
`refer to the area within the syringe that is sealed by the stopper 15
`from access from either end of the syringe. This may be the
`area between a seal of the stopper, for example a circumfer(cid:173)
`ential 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 sterility zone of 20
`the stopper since the stopper is installed into the syringe barrel
`in a sterile environment.
`To further assist in maintaining sterility during the opera(cid:173)
`tions noted above the stopper may comprise at a front circum(cid:173)
`ferential rib and a rear circumferential rib and those ribs may 25
`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 addi(cid:173)
`tional 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 30
`embodiment there are a total of three circumferential ribs.
`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(cid:173)
`conisation of the syringe, such an increase in silicone oil 40
`levels is particularly undesirable for syringes for ophthalmic
`use.
`The rod shoulder may be arranged within the external
`diameter of the rod, or may be arranged outside the external
`diameter of the rod. By providing a shoulder that extends
`beyond the external diameterofthe 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(cid:173)
`pendicular to the first axis. The rod shoulder may comprise
`any suitable shoulder forming elements on the rod, but in one 50
`embodiment the rod shoulder comprises a substantially disc
`shaped portion on the rod.
`In one embodiment of the syringe, when arranged with the
`plunger contact surface in contact with the stopper and the
`variable volume chamber is at its intended maximum volume 55
`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 ofless 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.
`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.
`In another embodiment the syringe is dimensioned so as to
`have a nominal maximum fill volume ofbetween about 0.1 ml
`
`4
`and about 1.5 ml. In certain embodiments the nominal maxi(cid:173)
`mum 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.
`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.
`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 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 wastage
`due to "dead space" within the syringe and needle. There may
`also be a certain amount of wastage when the syringe is
`primed by the physician, so that it is ready to inject the patient.
`Thus, in one embodiment, the syringe is filled with a dos(cid:173)
`age 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 or0.03 ml (0.5 mg or0.3 mg) ofa 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(cid:173)
`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(cid:173)
`ume from the syringe (that is the amount of product obtain(cid:173)
`able from the syringe following filling, taking into account
`35 loss due to dead space in the syringe and needle) is about 0.09
`ml.
`In one embodiment the length of the syringe body is
`between about 45 mm and about 50 mm, the internal diameter
`is between about 4 mm and about 5 mm, the fill 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.
`As the syringe contains a medicament solution, the outlet
`may be reversibly sealed to maintain sterility of the medica(cid:173)
`ment. This sealing may be achieved through the use of a
`45 sealing device as is known in the art. For example the OVS™
`system which is available from Vetter Pharma International
`GmbH.
`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(cid:173)
`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
`"floaters" and an increase in intra-ocular pressure. Further(cid:173)
`more, silicone oil can cause proteins to aggregate. A typical 1
`ml syringe comprises 100-800 µg silicone oil in the barrel,
`though a survey of manufacturers reported that 500-1000 µg
`was typically used in pre-filled syringes (Badkar et al. 2011,
`60 AAPS PharmaSciTech, 12(2):564-572). Thus,
`in one
`embodiment, a syringe according to the invention comprises
`less than about 800 µg (i.e. about less than about 500 µg, less
`than about 300 µg, less than about 200 µg, less than about 100
`µg, less than about 75 µg, less than about 50 µg, less than
`65 about 25 µg, less than about 15 µg, less than about 10 µg)
`silicone oil in the barrel. If the syringe comprises a low level
`of silicone oil, this may be more than about 1 µg, more than
`
`Regeneron Exhibit 1001.005
`
`

`

`US 9,220,631 B2
`
`5
`about 3 µg, more than about 5 µg, more than about 7 µg or
`more than about 10 µg silicone oil in the barrel. Thus, in one
`embodiment, the syringe may comprise about 1 µg-about 500
`µg, about 3 µg-about 200 µg, about 5 µg-about 10 µg or about
`10 µg-about 50 µg silicone oil in the barrel. Methods for 5
`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(cid:173)
`cone oil are available, but typically either DC360 (Dow Com- 10
`ing®; with a viscosity of 1000 cP) or DC365 emulsion (Dow
`Coming®; DC360 oil with a viscosity of350 cP) are used for
`syringe siliconisation. In one embodiment, the pre-filled
`syringe of the invention comprises DC365 emulsion.
`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 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. Schoe(cid:173)
`nknecht. AAPS National Biotechnology Conference 2007-
`Abstract no. NBC07-000488, which indicates that while 400 25
`µg silicone oil is acceptable, usability improves when
`increased to 800 µg). 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(cid:173)
`filled syringes contain about 100 µg-about 800 µg silicone oil.
`In one embodiment the glide/ slide force for the stopper within
`the pro-filled syringe is less than about 11 N or less than 9N,
`less than 7N, less than 5N or between about 3N to SN. In one
`embodiment, the break loose force is less than about 1 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 mea-
`sured at a stopper travelling speed of 190 mm/min. In one
`embodiment, the forces are measured with a 30 Gx0.5 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 µg silicone oil
`and has a break loose force between about 2N to SN.
`In one embodiment the syringe barrel has an internal coat(cid:173)
`ing 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,
`
`15
`
`20
`
`30
`
`35
`
`40
`
`45
`
`50
`
`6
`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®
`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-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 embodi(cid:173)
`ment, the ophthalmic solution comprises no more than 2
`particles 2:50 µmin diameter per ml. In one embodiment, the
`ophthalmic solution comprises no more than 5 particles 2:25
`µm in diameter per ml. In one embodiment, the ophthalmic
`solution comprises no more than 50 particles 2:10 µm in
`diameter per ml. In one embodiment, the ophthalmic solution
`comprises no more than 2 particles 2:50 µm in diameter per
`ml, no more than 5 particles 2:25 µmin diameter per ml and no
`more than 50 particles 2:10 µm 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
`VEGF is a well-characterised signal protein which stimu(cid:173)
`lates angiogenesis. Two antibody VEGF antagonists have
`been approved for human use, namely ranibizumab (Lucen(cid:173)
`tis®) and bevacizumab (Avastin®).
`Non-Antibody VEGF Antagonist
`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 (2007) Clin
`Cancer Res 13:4623-7s). Aflibercept is the preferred non(cid:173)
`antibody VEGF antagonist for use with the invention. Afliber(cid:173)
`cept 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 molecular mass,
`resulting in a total molecular weight of 115 kDa. It is conve(cid:173)
`niently produced as a glycoprotein by expression in recom(cid:173)
`binant CHO Kl cells. Each monomer can have the following
`amino acid sequence (SEQ ID NO: 1 ):
`
`SDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATY
`
`KEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPS
`
`SKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPP
`
`CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST
`
`YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK
`
`GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL
`
`SLSPG
`
`Regeneron Exhibit 1001.006
`
`

`

`US 9,220,631 B2
`
`7
`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-
`mers.
`Another non-antibody VEGF antagonist immunoadhesin
`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 protein fragment (Li et
`al., 2011 Molecular Vision 17:797-803). This antagonist
`binds to isoforms VEGF-A. VEGF-B and VEGF-C. The mol(cid:173)
`ecule is prepared using two different production processes
`resulting in different glycosylation patterns on the final pro(cid:173)
`teins. The two glycoforms are referred to as KH902 ( conber- 15
`cept) and KH906. The fusion protein can have the following
`amino acid sequence (SEQ ID N0:2):
`
`8
`be produced by the addition or deletion of amino acids. Ordi(cid:173)
`narily, these amino acid sequence variants will have an amino
`acid sequence having at least 60% amino acid sequence iden(cid:173)
`tity with the amino acid sequences of SEQ ID NO: 1, SEQ ID
`5 NO: 2 or SEQ ID 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
`10 100%. 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 ID NO: 1,
`SEQ ID NO: 2 or SEQ ID NO: 3, after aligning the sequences
`and introducing gaps, if necessary, to achieve the maximum
`percent sequence identity, and not considering any conserva(cid:173)
`tive substitutions as part of the sequence identity.
`
`MVSYWDTGVLLCALLSCLLLTGSSSGGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDT
`
`LIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEK
`
`LVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSG
`
`LMTKKNSTFVRVHEKPFVAFGSGMESLVEATVGERVRLPAKYLGYPPPEIKWYKNGIPLESNHTIKAGHVL
`
`TIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVYVPPGPGDKTHTCPLCPAPELLGGPSVFLFPPKPKDT
`
`LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
`
`KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLT