KR20220085906A - Syringe comprising ophthalmic formulation - Google Patents

Abstract

The present invention relates to a body (2) provided with an outlet (12) at an outlet end (14);
a plunger (4) located inside the body (2); and a front surface (16) located in the interior of the body (2) at the front of the plunger (4), which is silicon oil free and forms a variable volume chamber (18) with the body (2). and a stopper 10 for discharging the ophthalmic formulation 20 filled in the variable volume chamber 18 by the movement of the plunger 4 through the outlet 12, wherein the plunger 4 is , a plunger rod 26 extending in the longitudinal direction, and a flange portion 22 located at a first end 24 adjacent to the stopper 10 in the plunger rod 26 , and from the flange portion 22 . It includes a spherical or screw-shaped insertion fixing part 23 protruding toward the stopper 10, and an insertion fixing groove 62 into which the insertion fixing part 23 is inserted and fixed to the stopper 10 is provided. wherein the ophthalmic formulation comprises, in an aqueous medium, (a) aflibercept in a therapeutically effective amount of 10 to 100 mg/ml; (b) 2 to 12 w/v% of a stabilizer; (c) 0.01 to 0.1 w/v% of a surfactant; and (d) 5-100 mM of an ionic isotonic agent; It includes, has a pH of 5.2 to 6.2, and provides a silicone oil free syringe.

Description

Syringe comprising ophthalmic formulation

The present invention relates to a syringe containing an ophthalmic formulation, and more particularly, to a pre-filled syringe in which an ophthalmic formulation containing aflibercept as an active ingredient is filled.

A type of cell-derived dimer mitogen having selectivity for vascular endothelial cells has been identified, and it is termed vascular endothelial growth factor (VEGF). VEGF is an important factor that increases angiogenesis and vascular permeability.

VEGF is known to activate membrane-spanning tyrosine kinase receptors, VEGF receptors (VEGFR-1, VEGFR-2, VEGFR-3). Among the VEGF receptors, VEGFR-1 and VEGFR-2 have seven Ig-like sequences in the extracellular region for VEGF binding, a single transmembrane region, and a consensus tyrosine kinase region. This property is used as a sequence for a VEGF antagonist (anti-VEGF agent), and in aflibercept, an ophthalmic therapeutic agent, the second binding domain of VEGFR-1 and the third binding domain of VEGFR-2 are bound to the Fc region of human IgG1. It is a soluble decoy receptor of about 115 kDa (including glycosylation) in the structure of the present invention.

Aflibercept is a recombinant fusion protein composed of a VEGF binding site derived from the extracellular domains of human vascular endothelial growth factor (VEGF) receptors 1 and 2 fused to the Fc portion of human IgG1 immunoglobulin. to be. Aflibercept under the trade name Eylea ^™ has been approved in the United States and Europe for the treatment of wet macular degeneration. Abnormal angiogenesis in the mechanism through VEGF is ophthalmic diseases wet age-related macular degeneration, diabetic macular edema, and retinal vein occlusion-macular edema (macular edema in retinal vein). occlusion) are known to be related.

The protein-containing pharmaceutical composition is physicochemically denatured under conditions that are not optimal. In particular, factors such as concentration of protein, type of buffer, type and concentration of stabilizer, type and concentration of organic cosolvent, concentration of salt, pH, temperature, contact with air, etc. can cause protein oxidation and deamidation. (deamidation), isomerization (isomerization), polymerization (polymerization), etc. have a significant effect. Such denaturation may reduce physiological activity by generating protein aggregation, fragments, and isomers. In particular, protein aggregation is a major intrinsic factor in the formation of insoluble particles, which can cause side effects such as immune responses. In addition, due to the specificity of the ophthalmic formulation, the standards for insoluble microparticles should be maintained more strictly.

Accordingly, although the development of an ophthalmic formulation containing aflibercept, a recombinant fusion protein composed of a VEGF binding site, has been actively conducted, conventional formulations have a problem in that the stabilizing effect of aflibercept is reduced under severe conditions.

On the other hand, many medicaments are delivered to a patient from a syringe through which a user can dispense the medicament, and when the medicament is delivered to the patient by means of a syringe, it is common to enable the patient or caregiver to inject the medicament. It is important for patient safety and medicinal integrity that the syringe and its contents are sufficiently sterile to avoid infection or other risks to the patient. Sterilization may be accomplished by terminal sterilization, typically by sterilizing the assembled product already placed in its associated packaging with heat or sterilizing gas.

For small volume syringes, eg syringes for injection into the eye intended to inject less than about 0.1 ml of liquid, sterilization can present difficulties that are not necessarily associated with large syringes. Changes in the internal or external pressure of a syringe can cause parts of the syringe to move unpredictably, which can change the sealing properties and potentially compromise sterility. In addition, incorrect handling, including assembling syringes, may pose a risk to product sterilization.

Therefore, there is a need to develop an ophthalmic formulation containing aflibercept that can be stably present even under severe conditions, and a syringe that safely delivers it to the eye and has excellent storage stability.

Accordingly, the present inventors have found that a fusion protein such as Aflibercept is not only stable for a long period of time under storage conditions, but also a liquid formulation with improved stability even under harsh conditions, and the formulation can be safely delivered to the eye, and the existing commercially available ophthalmic syringe The present invention has been completed by developing a syringe that has more improved properties and can stably exist during storage.

The present invention provides a silicone-free syringe filled with an ophthalmic formulation containing aflibercept as an active ingredient in order to solve the above problems.

Specifically, an object of the present invention is a body (2) provided with an outlet (12) at the outlet end (14);

a plunger (4) located inside the body (2); and

Located in the interior of the body (2) at the front of the plunger (4), it is silicon oil free and has a front surface (16) forming a variable volume chamber (18) with the body (2). and a stopper 10 for discharging the ophthalmic formulation 20 filled in the variable volume chamber 18 through the outlet 12 by the movement of the plunger 4,

The plunger (4) includes a plunger rod (26) extending in the longitudinal direction, and a flange portion (22) located at a first end (24) of the plunger rod (26) adjacent to the stopper (10); Includes a spherical or screw-shaped insertion fixing part 23 protruding from the flange part 22 toward the stopper 10,

An insertion fixing groove 62 into which the insertion fixing part 23 is inserted and fixed is provided in the stopper 10,

The ophthalmic formulation comprises, in an aqueous medium, (a) a therapeutically effective amount of aflibercept; (b) stabilizers; (c) surfactants; and (d) an ionic isotonic agent; It contains, has a pH of 5.2 to 6.2, to provide a silicone-free (silicon oil free) syringe.

Another object of the present invention is to provide a method for filling the syringe with an ophthalmic formulation.

The present invention relates to a body (2) provided with an outlet (12) at an outlet end (14);

a plunger (4) located inside the body (2); and

Located in the interior of the body (2) at the front of the plunger (4), it is silicon oil free and has a front surface (16) forming a variable volume chamber (18) with the body (2). and a stopper 10 for discharging the ophthalmic formulation 20 filled in the variable volume chamber 18 through the outlet 12 by the movement of the plunger 4,

The plunger (4) includes a plunger rod (26) extending in the longitudinal direction, and a flange portion (22) located at a first end (24) of the plunger rod (26) adjacent to the stopper (10); Includes a spherical or screw-shaped insertion fixing part 23 protruding from the flange part 22 toward the stopper 10,

An insertion fixing groove 62 into which the insertion fixing part 23 is inserted and fixed is provided in the stopper 10,

The ophthalmic formulation may contain, in an aqueous medium, (a) aflibercept in a therapeutically effective amount from 10 to 100 mg/ml; (b) 2 to 10 w/v% of a stabilizer; (c) 0.01 to 0.1 w/v% of a surfactant; and (d) 5-100 mM of an ionic isotonic agent; It contains, has a pH of 5.2 to 6.2, and provides a silicone oil free syringe.

The syringe of the present invention is for use in administering an ophthalmic formulation to a patient with an ophthalmic disease, wherein the ophthalmic disease is age-related macular degeneration (AMD), visual impairment due to diabetic macular edema (DME), retinal vein occlusion (branch visual impairment due to secondary macular edema of RVO or central RVO), diabetic retinopathy in patients with diabetic macular edema, or visual impairment due to secondary choroidal neovascularization (CNV) in pathologic myopia.

Hereinafter, the present invention will be described in more detail.

Syringes containing ophthalmic formulations

The syringe of the present invention includes a body (2) provided with an outlet (12) at an outlet end (14); a plunger (4) located inside the body (2); and a front surface (16) located in the interior of the body (2) at the front of the plunger (4), which is silicon oil free and forms a variable volume chamber (18) with the body (2). and a stopper 10 for discharging the ophthalmic formulation 20 filled in the variable volume chamber 18 by the movement of the plunger 4 through the outlet 12, wherein the plunger 4 is , a plunger rod 26 extending in the longitudinal direction, and a flange portion 22 located at a first end 24 adjacent to the stopper 10 in the plunger rod 26 , and from the flange portion 22 . It includes an insertion fixing part 23 in the form of a spherical or screw that protrudes toward the stopper 10, and the insertion fixing part 23 is pressed into the stopper 10, and the insertion fixing groove 62 is inserted and fixed. ) may be provided. Here, the screw shape means a screw shape with a partially protruding surface, and a spherical shape means that it does not have a partially protruded surface and has a smooth shape, unlike the screw shape, and preferably has a bore 64 of the insertion fixing groove 62 . A lower end portion 64 ′ having a size and shape similar to or identical to that of the inner fixing groove 66 may have an upper end portion 66 ′ having a size and shape similar or identical to that of the inner fixing groove 66 .

In the syringe of the present invention, as the plunger and the stopper are coupled and connected by a press-fit coupling of the insertion fixing part 23 to the insertion fixing groove 62 of the stopper 10, the plunger and the stopper are integrated into the stopper has the advantage of being retractable, not distorted in an unpredictable manner, without detrimental to the sealing and/or sterilization of the final product, reliably storing the ophthalmic formulation without leakage or contamination, and providing adequate delivery of the ophthalmic formulation. . In addition, if the plunger and the stopper are integrally coupled, the plunger shoulder 32 is configured at a specific position of the plunger body, and the backward prevention shoulder 34 is included, the air bubbles generated in the filling process will be removed from the subsequent steam hydrogen peroxide sterilization process or ethylene oxide. It is possible to prevent problems such as unexpected sterilization cracking because it has the advantage of being able to control so that the stopper does not retreat from the intended position more precisely to retract the stopper while expanding under the reduced pressure accompanying the seed gas sterilization process.

In the syringe of the present invention, the body 2 may be a substantially cylindrical shell, or may include a substantially cylindrical bore 36 having a non-circular outer shape. The outlet end 14 of the body 2 comprises an outlet 12 from which the ophthalmic formulation 20 stored in the variable volume chamber 18 can be released when the volume of the variable volume chamber 18 is reduced. The outlet 12 may include a protrusion from the outlet end 14 through which a channel having a diameter less than the diameter of the variable volume chamber 18 extends. The outlet 12 may be configured to connect to a needle or other accessory, such as a sealing device, for example by a luer lock type connection, which may seal the variable volume chamber but may be manipulated or removed to close the variable chamber. It may be opened to allow connection of the syringe with another accessory, such as a needle. This connection may be made directly between the syringe and the accessory or by a sealing device. The body 2 extends along a first axis A from the outlet end 14 to the rear portion 25 .

The body 2 may be made from a plastic material or from glass or from any other suitable material and may include on its surface an indicator serving as an injection guide. The plastic material may be, for example, a cycloolefin polymer (COP), a cycloolefin copolymer (COC), or the like. Preferably, a COP material is used.

The stopper 10 may be made of a deformable material such that the stopper body 58 has rubber, silicone, or other suitable elastic and deformable material. The stopper 10 provides a sealing function by forming a leak-tight seal at the rear of the variable volume chamber for the ophthalmic formulation 20 filled in the variable volume chamber. The stopper 10 may be substantially cylindrical, and the stopper may include one or more ribs 52, 56 around an outer surface of the stopper, wherein the stopper 10 and the ribs 52, 56 are ribbed. It may contact the inner surface of the syringe body to form a substantially leak-tight seal. The front surface 16 of the stopper may have any suitable shape and may be planar or conical, preferably conical. In addition, the stopper may include an insertion fixing groove 62, and the insertion fixing groove 62 may have any shape as long as it does not harm the sealing function of the stopper. The insertion groove 62 may be cylindrical, or the insertion groove 62 may include a bore 64 having a first diameter, the bore 64 being a length within the stopper from the rear surface 60 . It leads to an inner fixing groove 66 having a second diameter extending in the direction, the second diameter being larger than the first diameter and may be conical. By inserting the insertion fixing part 23 of the plunger into the insertion fixing groove 62, the stopper and the plunger may be coupled. In addition, the insertion fixing part 23 includes a spherical shape or a screw shape, and may preferably have a spherical shape. The spherical insertion fixing part 23 can be strongly fixed to the stopper by the spherical protrusion of the insertion fixing part 62 while being press-fitted into the insertion fixing groove 62 of the stopper having elasticity. Compared to the additional screw type, there is an advantage that can be stably coupled without damaging the stopper to be coupled ( FIGS. 7 and 8 ). Therefore, it is more preferable that the shape of the insertion fixing part is spherical. In addition, removing material from the central portion of the stopper (if this is not necessary for the stopper to perform its required function) reduces the stopper weight and reduces the amount of material required to manufacture the stopper while reducing the insertion fixture 23 of the plunger. ) may be coupled by being inserted into the insertion fixing groove 62 of the stopper.

The plunger (4) includes a flange portion (22) and a plunger rod (26) extending from the flange portion (22) to a rear portion (25). The rear portion 25 may include a disc-shaped flange 42 which is a user-contacting portion adapted to be contacted by a user while the formulation filled in the syringe is being injected. The disk-shaped flange 42 may include a disk-shaped portion, the radius of the disk extending perpendicular to the axis along which the rod extends, but the user-contacting portion may have a variety of other suitable shapes other than the disk shape. The axis along which the plunger rod 26 extends may be the first axis A and may be substantially parallel to the first axis.

The flange part 22 is positioned at the first end 24 adjacent to the stopper 10 and may have a structure capable of being coupled to the stopper 10 . Specifically, by including a spherical or screw-shaped, preferably spherical-shaped insertion fixing part 23 protruding from the central portion of the flange part 22 toward the stopper, the insertion fixing part 23 is the insertion fixing groove of the stopper. The plunger 4 and the stopper 10 can be coupled by being inserted into the 62 by a pressing action, and the coupled form is as shown in FIGS. 7 and 8 (right side of FIGS. 7 and 8, respectively). In addition, the flange portion 22 may have an outer diameter smaller than the inner diameter of the body 2 and may have a circular shape. Also, the diameter of the flange portion 22 may be the same as the diameter of the rear surface 60 of the stopper in contact with the flange portion 22 . Also, the flange portion 22 may be rotationally symmetrical to the rear surface 60 of the stopper. In the present invention, since the stopper 10 and the plunger 4 are coupled and mechanically connected, the plunger and the stopper form an integral body, and the stopper can be moved together to the rear of the syringe according to the movement of the plunger. When the stopper and the plunger are integrated in this way, the air bubbles generated in the filling process expand under reduced pressure accompanying the subsequent steam hydrogen peroxide sterilization process or ethylene oxide gas sterilization process, and are no longer at the position where the stopper is more accurately intended to retract. It has the advantage of being able to control not to retreat, so it is possible to prevent problems such as unexpected sterilization cracking.

The plunger rod 26 may have a circular or cross-section. The cross-section may be formed from a plunger rib 44 extending along at least a portion of the plunger rod 26 , the plunger rod 26 including a disc-shaped portion 46 , the disc-shaped portion 46 including the plunger It may extend radially beyond the ribs 44 .

The plunger rib 44 may extend substantially parallel to an axis along which the plunger rod 26 extends, and the cross-section may provide rigidity to the plunger rod 26 without increasing manufacturing complexity. In addition, the plunger rod 26 may be made of a plastic such as cycloolefin polymer (COP) and cycloolefin copolymer (COC), preferably made of cycloolefin polymer (COP). The plunger rod 26 may be substantially rigid under anticipated conditions of use.

The syringe may comprise an anti-retraction part 6 arranged on the rear part of the body. The anti-retraction part 6 may be removable from the syringe. When the body (2) of the syringe includes a distal flange (28) at the end opposite to the outlet end (14), the anti-retraction (6) prevents the distal flange (28) of the body (28) from sandwiching the sandwich portion (30). It may be configured to be positioned, which may prevent movement of the anti-reverse portion 6 in a direction parallel to the first axis A.

The plunger 4 may also comprise at least one plunger shoulder 32 positioned on the portion of the plunger rod 26 opposite the first end 24 . The plunger shoulder 32 may be arranged within the outer diameter of the plunger rod 26 or may be arranged outside the outer diameter of the plunger rod 26 . The plunger shoulder 32 extends beyond the outer diameter of the plunger rod 26 but still fits within the body and reduces the movement of the plunger rod 26 perpendicular to the first axis A, thereby reducing the plunger within the body 2 . It may serve to stabilize the movement of the rod (26).

Further, the anti-backward portion 6 may be attached to the body 2 by coupling to the distal flange 28 of the body 2, wherein at least a portion of the distal flange 28 of the body 2 is substantially sanded. may include a sandwich portion (30) adapted to be positioned by leaving one side of the anti-retraction portion (6) open so that the anti-retraction portion (6) can be fitted into the body (2) on that side of the body It can be adapted to couple to (2) (FIG. 10).

In addition, the anti-retraction portion 6 includes two finger projections 40 perpendicular to the first axis A and extending in opposite directions away from the body 2 to facilitate the use and handling of the syringe 1 . can be done easily

In addition, the anti-reverse part 6 is located on the rear part 25 of the body 2 to prevent the plunger 4 from being separated by cooperation with the plunger shoulder 32 when the plunger 4 moves backward. Solder 34 for preventing backward movement may be included. Restricting movement of the plunger rod 26 away from the outlet end may help maintain sterility during terminal sterilization operations, or other operations where the pressure may vary within or outside the variable volume chamber.

Also, the plunger rib 44 may be substantially hollow and may include a void 48 . The disc-shaped portion 46 may be hollow or may include voids 50 . The voids 48 and 50 may be used to facilitate gas flow within the body 2 for sterilization or other purposes, if necessary, but such voids are not necessarily included and may be added and removed as appropriate. have.

As mentioned above, the syringe may be sterilized using terminal sterilization methods, which may use known methods such as ethylene oxide or steam hydrogen peroxide sterilization methods.

This sterilization process has a problem in that air bubbles generated in the filling process expand under the reduced pressure accompanying the sterilization process, resulting in a change in volume in the syringe. On the other hand, the combination of the stopper and the plunger according to the present invention has the advantage of being able to control the volume change more precisely at the intended position, so that it is possible to prevent problems such as unexpected sterilization cracking.

The inclusion of one or more circumferential ribs 52 , 56 on the stopper 10 may alter the force required to move the stopper from its rest position, and may alter the sealing properties of the stopper. The stopper may also serve to protect the injectable medicament (ophthalmic formulation) during terminal sterilization methods. Since some medicaments, such as biopharmaceuticals, can be damaged by exposure to sterilizing agents such as ethylene oxide, the ribs of the stopper can reduce this exposure and thus protect the medicament filled in the syringe. By adjusting the number of the ribs and the distance between the ribs, exposure and damage by the sterilizing agent can be reduced.

The sealing part 8 is composed of an upper end 81 and a lower end 82, and the lower end may have a smaller diameter than the upper end, and the lower end 82 has a hollow 83 that is hollow in the center. can have An outlet 12 provided at the outlet end 14 of the body 2 may be inserted into the depression 83 , and the distal end of the outlet contacts and seals the protrusion 84 to seal the formulation 20 filled in the syringe. ) can be prevented from leaking from the outlet (12). In addition, the lower end 83 is a part that can be coupled to the body 2 and includes a screw shape, so that it can be sealed by a screw tightening action. A part of the body 2 coupled to the lower end may also include a screw shape. The upper end 81 of the sealing part 8 is a part that the user contacts to release the sealing by the coupling of the lower end and the body, and may have a shape that repeatedly protrudes at regular intervals to prevent slipping (Fig. 9).

According to the present invention, the syringe is suitable for ophthalmic injection and thus has a suitably small volume. The syringe may be configured for ophthalmic injection. In addition, the syringe does not contain silicone oil, and preferably, the stopper of the present invention is silicone-free, and at least the surface of the stopper in contact with the drug solution, more preferably the entire stopper is not coated with silicone oil. However, other lubricants that facilitate movement of the stopper may be included instead of silicone oil.

The syringe of the present invention is an ophthalmic syringe in which the variable volume chamber is filled with an injectable ophthalmic formulation and the outlet can be reversibly sealed.

In addition, the variable volume chamber of the syringe may be filled with any suitable injectable liquid or medicament, eg, may be filled with an injectable medicament. The variable volume chamber has an inner diameter greater than 5 mm or 6 mm and less than 3 mm or 4 mm. The inner diameter may be between 3 mm and 6 mm, or between 4 mm and 5 mm. Also, the volume of the liquid formulation that can be filled into the variable volume chamber is from about 0.05 ml to about 1 ml, preferably from about 0.1 ml to about 0.5 ml, more preferably from 0.14 ml to 0.3 ml, most preferably from 0.15 ml to 0.2 ml.

Those skilled in the art know that syringes are generally filled to a larger volume than the volume that will actually be administered to the patient, taking into account the losses due to the manufacture of injectable syringes. Therefore, the volume actually administered to the patient is 0.01 ml to 1 ml, preferably 0.02 to 0.5 ml, more preferably 0.025 to 0.3 ml, even more preferably 0.03 ml to 0.1 ml.

The syringe of the present invention is shown in Figs. 1 to 3, the plunger is shown in Figs. 4 to 6, the stopper is shown in Figs. 7 and 8, the sealing unit is shown in Fig. 9, and the reverse preventing unit is shown in Fig. 10.

In the method of assembling a syringe and filling the syringe with the ophthalmic formulation of the present invention, the stopper of the present invention can be mounted aseptically by an automatic filling machine after the ophthalmic formulation is filled in the variable volume chamber.

Specifically, the method of filling the syringe of the present invention with the ophthalmic formulation comprises the steps of: 1) preparing the ophthalmic formulation of claim 1; 2) sterilizing and filtering the prepared ophthalmic formulation; 3) aseptically filling the sterilized and filtered formulation into the variable volume chamber of a syringe using a pump of an automatic filling machine; 4) aseptically mounting the stopper by an automatic charger; and 5) coupling the stopper with the plunger.

Ophthalmic formulations filled in syringes

The ophthalmic formulations of the present invention may be filled into the variable volume chamber of a syringe.

The ophthalmic formulations according to the invention have formulations most suitable for use in particular in the above-mentioned syringe conditions. Specifically, when the silicone oil is not coated on the inside of the syringe, in general, the active ingredient in the formulation may cause a problem in stability, such as adsorption or decomposition on the inner wall. However, the ophthalmic formulation according to the present invention is a formulation capable of ensuring excellent stability even in a syringe without such silicone oil, and may exhibit excellent properties when provided with a gastric syringe.

The ophthalmic formulation comprises, in an aqueous medium, (a) a therapeutically effective amount of aflibercept; (b) stabilizers; (c) surfactants; and (d) an ionic isotonic agent; and may have a pH of 5.2 to 6.2.

In the present invention, the ophthalmic formulation comprises (a) aflibercept in a therapeutically effective amount of 10 to 100 mg/ml; (b) 2 to 10 w/v% of a stabilizer; (c) 0.01 to 0.1 w/v% of a surfactant; and (d) 5-100 mM of an ionic isotonic agent; Including, and may be one having a pH of 5.2 to 6.2.

In the present invention, aflibercept is a VEGF-specific fusion protein in which a VEGF binding site derived from the extracellular domains of human VEGF receptor 1 and VEGF receptor 2 and an Fc region of human IgG1 are fused. Specifically, the VEGF-specific fusion protein comprises a region essentially comprising immunoglobulin-like (Ig) domain 2 of human VEGF receptor 1 (Flt1) and Ig domain 3 of human VEGF receptor 2 (Flt1 or Flt4) and human IgG1 It is a fusion protein in which the Fc region is fused, and may be aflibercept having the amino acid sequence of SEQ ID NO: 1 below.

The amino acid sequence of aflibercept (SEQ ID NO: 1) is as follows:

(Disulfide bridge: 30-79; 124-185; 246-306; 352-410, Dimer: 211; 214)

Such aflibercept may be recombinantly or synthetically produced.

In the present invention, "excellent stability" or "kept stable" may mean that the structure and/or physical, chemical, and/or biological properties of a protein in the composition are maintained during storage (eg, storage low protein duplex, multimer, polymer formation rate, low protein aggregation rate, low protein fragment production, degradation rate, and/or low denaturation rate, etc.). Various analytical techniques for measuring the stability of proteins are well known in the art. The protein shows little or no change in aggregation, precipitation and/or denaturation when observed by visual inspection of color and/or clarity, or as determined by UV light scattering (to measure visible aggregates) or size exclusion chromatography (SEC). If there is no change at all, it can be said that the formulation "maintains its physical stability".

The ophthalmic formulation of the present invention may contain aflibercept in an amount of 10 to 100 mg/ml, preferably 10 to 50 mg/ml, more preferably 20 to 50 mg/ml as a therapeutically effective amount of aflibercept. can do.

In the present invention, the ophthalmic formulation may further include a buffer of 5 to 20 mM, and the concentration of the buffer may be preferably 7 to 15 mM, more preferably 8 to 12 mM. . A “buffered solution” refers to a buffered solution that withstands changes in pH due to the action of its acid-base conjugate component. In one embodiment of the present invention, the buffer solution may be an acetate buffer solution, preferably sodium acetate buffer solution.

The aqueous medium includes, without limitation, an aqueous medium capable of providing a buffer together with an acetate salt buffer, and includes, for example, distilled water for injection, sterile purified water, and the like.

In the present invention, the stabilizer may include sucrose, trehalose, mannitol, glucose, and the like, preferably sucrose. In addition, the concentration of the stabilizer may be 2 to 12 w/v%, preferably 3 to 10 w/v%, more preferably 5 to 8 w/v%.

In the present invention, the surfactant may include polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, etc., preferably polysorbate 20 may be used. In addition, the formulation may contain a surfactant in a concentration of 0.01 to 0.1 w/v%, preferably 0.01 to 0.08 w/v%, more preferably 0.01 to 0.05 w/v%.

In the present invention, the ionic tonicity agent is sodium chloride, and the formulation of the present invention contains sodium chloride, an ionic tonicity agent, of 5 to 100 mM, preferably 10 to 50 mM, more preferably 10 to 40 mM. concentration may be included.

The formulation provided herein may be isotonic with the living body. For example, the formulation may have an osmotic pressure of about 200 mOsm/kg to about 400 mOsm/kg, for example, about 250 mOsm/kg to about 300 mOsm/kg, the scope of the present invention is not limited thereto, and the osmotic pressure can be controlled by stabilizing agents.

An ophthalmic application according to the present invention (a) a therapeutically effective amount of 20 to 50 mg/ml of aflibercept; (b) 8-12 mM sodium acetate buffer; (c) 5 to 8 w/v% sucrose; (d) 0.01 to 0.05 w/v% polysorbate 20; and (e) 10-40 mM of an ionic isotonic agent of sodium chloride; may include, and the pH of the formulation may be 5.4 to 6.2, preferably, 5.4 to 6.0.

More specifically, an ophthalmic application according to the present invention (a) a therapeutically effective amount of about 40 mg/ml of aflibercept; (b) about 10 mM sodium acetate buffer; (c) about 6.5 w/v% sucrose; (d) 0.03 w/v% polysorbate 20; and (e) about 15 mM of an ionic isotonic agent of sodium chloride; may include, and may be 5.4 to 6.0. Specifically, the pH may be about 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 or 6.0.

The ophthalmic formulation may be provided in the form of a syringe of the present invention in the form of a prefilled syringe.

The ophthalmic formulation according to the present invention has excellent self-stability, and exhibits excellent effects of inhibiting the formation of aggregates (dimers, multimers, etc.) and fragments.

In another aspect of the present invention, the present invention also provides a kit comprising one or more prefilled syringes of the present invention. Preferably, the kit comprises a blister pack. A "blister pack" has a cavity or pocket, usually made of thermoformed plastic, and a backing of cardboard or a lidding seal of aluminum foil or Tyvek material. The blister pack can be sterilized inside the blister pack using steam hydrogen peroxide or ethylene oxide gas after sealing the Tyvek lid. The kit may further include a needle if the prefilled syringe does not include a stack-in needle. The kit may further include instructions for use.

The syringe containing the ophthalmic formulation of the present invention has excellent storage, and can maintain the efficacy of the drug even when stored for a long period of time. Since the structure of the syringe is also stable, it can be safely delivered to the eye without leakage of the formulation, and the formulation can be stably present in the syringe during the storage period of the formulation.

1 is a view showing the side of the syringe of the present invention.
2 and 3 are views showing a cross-section of the syringe of the present invention.
4 and 5 are views showing the plunger of the present invention.
6 is a view showing a cross-section of the plunger.
7 and 8 are views showing a cross-section of a stopper and a cross-section in which the plunger and the stopper are coupled.
9 is a view showing a side and a cross-section of a sealing part.
10 is a view showing a reverse preventing unit.

Hereinafter, the present invention will be described in detail by way of Examples. However, the following examples only illustrate the present invention, and the present invention is not limited by the following examples.

Example 1: Preparation of ophthalmic formulations

An ophthalmic formulation was prepared according to the ingredients and contents of Table 1 below.

The pH of the formulation was adjusted to be 5.2 to 6.2 by changing the ratio of sodium to acetate in the sodium acetate buffer, and in the case of polysorbate 20, it was added to a content of 0.03% at the end, as shown in Table 1 below, about 40 mg/ A liquid formulation containing aflibercept in ml content was prepared.

division ingredient content active ingredient aflibercept 40 mg/ml Acetate Buffer sodium acetate 10 mM stabilizer sucrose 6.5% Surfactants Polysorbate 20 0.03% ionic isotonic agents sodium chloride 15 mM menstruum water remaining amount

Example 2: Preparation of syringe formulations filled with ophthalmic formulations

The ophthalmic formulation prepared in Example 1 was sterilized and filtered using a disposable 0.22 μm polyethersulfone membrane. Thereafter, the syringe according to the present invention was aseptically filled with a volume of about 0.17 ml into the variable volume chamber by using the pump of the automatic filling machine.

The above-mentioned syringe has a silicone oil-free body made of COP material. A syringe filled with the ophthalmic formulation of Example 1 (specifically, a formulation having a pH of 5.5 is used) by coupling the stopper and the plunger and mounting the anti-reverse part after the stopper is aseptically mounted by the automatic filling machine after filling The formulation was prepared.

Example 3: Analysis of stability under refrigerated storage conditions and accelerated storage conditions of the syringe formulation of the present invention

In the refrigerated storage conditions (5 ± 3 °C) and accelerated storage conditions (25 ± 2 °C / 60 ± 5% RH) of the syringe formulation prepared in Example 2 (specifically, a formulation having a pH of 5.5 is used) Stability was confirmed, and for this purpose, analysis of insoluble particles under each condition, analysis of binding ability to VEGF, evaluation of fragment/dimer/multimer production, asparagine deamidation analysis, electrophoresis analysis, and biological activity analysis were performed, and these The analysis method is as follows.

(1) Method for analysis of insoluble particles

In order to analyze the insoluble particles that may be generated during the stability test period, it was analyzed by the method of USP<789> using the HIAC9703+ (Beckman Coulter) instrument. It was evaluated whether particles of μm or larger satisfy 5 or less per 1 mL, and the results are shown in Tables 2 and 3.

(2) VEGF Binding potency evaluation method

To evaluate the binding ability of aflibercept to VEGF, 200 ng/mL of rhVEGF was coated on a plate and incubated at 4 °C for 18 to 20 hours. After blocking for 1 hour, 1000, 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78, 0.39, 0.08 ng/mL of a standard solution (Eylea) and a test solution were loaded and at room temperature at 200 rpm for 1 hour. reacted. Detection antibody (Goat anti-Human IgG Fc Cross Adsorbed Secondary Antibody, HRP conjugate) diluted with 1/10,000, TMB substrate solution was sequentially treated for 1 hour and 30 minutes to react.

After completion of the reaction by adding 1 M sulfuric acid, the absorbance (OD) was measured at 450 nm using an ELISA Reader (Molecular Devices, SpectraMax M3) to obtain the EC50 value of the sample. It is summarized in 3.

(3) size exclusion HPLC analysis fragments used, dimer , and multimer Generating evaluation method

Size exclusion chromatography (SEC-HPLC) is used to determine the presence and content of various impurities such as higher molecular weight dimers and multimers or fragments of protein material that may be generated during stability testing. The peak area % of the main peak (monomer) and the peak area % of the measured impurity were obtained, and the analysis conditions for size exclusion HPLC are as follows:

- column; TSK-GEL G3000SWXL, 7.8 X 30 (ID mm X L cm) (Tosoh, Cat. # 08541), mobile phase: 200 mM potassium phosphate buffer (pH 6.2) containing 250 mM KCl, flow rate: 0.5 ml/min, wavelength: 280 nm.

The results of size exclusion HPLC analysis under the above conditions are summarized in Tables 2 and 3.

(4) asparagine deamidation ( Asn deamidation ) analysis method

Protein denaturation by performing asparagine deamidation analysis, that is, quantitative analysis of isoaspartate for asparagine deamidation quantification, on samples for stability testing stored under refrigerated conditions (2-8°C) and accelerated conditions (25°C). was evaluated. Asparagine deamidation analysis was performed using the ISOQUANT Isoasparatate Detection Kit (Promega, Cat. # MA1010) according to the manufacturer's instructions, and the results are summarized in Tables 2 and 3.

(5) isoelectric focusing ( Isoelectric focusing) electrophoretic analysis method

To check the protein isoelectric point pattern, 10 μg of standard and test solution are prepared, loaded into Novex® pH 3-10 IEF Gel 1.0 mm, 10 Well (Life technologies), and each at 100 V, 200 V, and 500 V voltage for 1 hour. , 1 hour, 30 minutes electrophoresis was performed. The gel was fixed in 12% trichloroacetic acid for 30 minutes, washed 3 times for 5 minutes each with ultrapure water, and treated for 1 hour until sufficiently stained with Instant Blue dye (Expedeon, Cat. # 1SB1L). After staining was completed, the isoelectric band was visually confirmed by washing with ultrapure water, and the intensity of each isoelectric band was analyzed using imaging equipment, and the results are shown in Tables 2 and 3.

(6) cell proliferation deterrent Biological activity evaluation method through analysis

For the ophthalmic formulation included in the syringe of the present invention, the biological activity under refrigerated storage conditions and accelerated conditions was evaluated.

That is, when umbilical vein endothelial cells (HUVEC, Human Umbilical Vein Endothelial Cells) are treated with vascular endothelial growth factor (VEGF, Vascular Endothelial Growth Factor), the cells proliferate. At this time, an in vitro test was performed to evaluate the degree of inhibition of cell proliferation caused by treatment with aflibercept.

In this test, the final treatment concentration of rhVEGF165 (Promokine, Cat. # C-64420) is 50 ng/ml, and aflibercept is set to have a concentration gradient of 2.4 to 2500 ng/ml at 37°C for 3 hours, Cells were treated with samples subjected to a neutralization reaction in a humidified 5% CO ₂ incubator, and then cultured for 3 days. In order to evaluate the degree of inhibition of cell proliferation, alamarBlue (Invitrogen Cat. # DAL 1025) reagent was added and incubated for another 14 to 20 hours. After shaking the 96-well plate at 200 rpm for 30 minutes before analysis, the concentration (IC ₅₀ ) of the drug required for half (50%) inhibition was measured by measuring the fluorescence intensity at 590 nm, and the results are shown in Table 2 and Table 3 summarized.

(7) Results

The stability test results of the syringe formulation prepared in Example 2 under refrigerated storage conditions (5 ± 3 °C) are shown in Table 2 below.

Test Items standard Example 2 (syringe formulation) 0 months 1 month 3 months 6 months appearance Transparent syringe with colorless or light yellow transparent liquid pass pass pass pass particle Free of insoluble foreign matter pass pass pass pass pH 5.4 - 6.0 5.8 6.0 5.7 5.8 osmotic pressure 270 ± 30 mOsm/kg 278 277 276 277 insoluble particles ≥10 μm: ≤50 counts/mL
≥25 μm: ≤5 counts/mL ≥10 μm: 3
≥25 μm: 1 ≥10 μm: 16
≥25 μm: 1 ≥10 μm: 7
≥25 μm: 0 ≥10 μm: 9
≥25 μm: 0 Molecular Weight (SDS-PAGE) Band shape equivalent to standard product pass pass pass pass VEGF-A binding capacity EC50 value is within 70 - 130% of standard product 90% 92% 89% 94% Protein content (UV 280nm) 40 ± 4 mg/mL 40 39 40 40 Size exclusion HPLC Main peak: 95% Excess aggregate: 5% or less Main peak: 99%
Aggregate: 1% Main peak: 99%
Aggregate: 1% Main peak: 98%
Aggregate: 2% Main peak: 98%
Aggregate: 2% Asparagine deamidation 0.76 pmol/pmol or less <0.35 pmol/pmol <0.35 pmol/pmol <0.35 pmol/pmol <0.35 pmol/pmol isoelectric focusing electrophoresis (IEF) 1. The sum of the band strengths in the range of isoelectric point 7,0-7.7 is 80% or more2. No new bands with isoelectric points above 8.2 and below 6.5 1. 93%.
2. pass 1. 90%.
2. pass 1. 87%.
2. pass 1. 92%.
2. pass biological activity
(HUVEC cell proliferation inhibitory ability) IC50 value is within 70 - 130% of standard product 91% 98% 90% 97%

The stability test results of the syringe formulation prepared in Example 2 under accelerated storage conditions (25 ± 2 °C / 60 ± 5% RH) are shown in Table 3 below.

Test Items standard Example 2 (syringe formulation) 0 months 1 month 3 months 6 months appearance Transparent syringe with colorless or light yellow transparent liquid pass pass pass pass particle Free of insoluble foreign matter pass pass pass pass pH 5.4 - 6.0 5.8 6.0 5.7 5.7 osmotic pressure 270 ± 30 mOsm/kg 278 281 278 277 insoluble particles ≥10 μm: ≤50 counts/mL≥25 μm: ≤5 counts/mL ≥10 μm: 3
≥25 μm: 1 ≥10 μm: 9
≥25 μm: 1 ≥10 μm: 8
≥25 μm: 0 ≥10 μm: 7
≥25 μm: 1 Molecular Weight (SDS-PAGE) Band shape equivalent to standard product pass pass pass pass VEGF-A binding capacity EC50 value is within 70 - 130% of standard product 90% 93% 81% 77% Protein content (UV 280nm) 40 ± 4 mg/mL 40 39 41 40 Size exclusion HPLC Main peak: 95% Excess aggregate: 5% or less Main peak: 99%
Aggregate: 1% Main peak: 98%
Aggregate: 2% Main peak: 97%
Aggregate: 3% Main peak: 97%
Aggregate: 3% Asparagine deamidation 0.76 pmol/pmol or less <0.35 pmol/pmol <0.35 pmol/pmol <0.35 pmol/pmol <0.35 pmol/pmol isoelectric focusing electrophoresis (IEF) 1. The sum of the band strengths in the range of isoelectric point 7,0-7.7 is 80% or more2. No new bands with isoelectric points above 8.2 and below 6.5 1. 93%.
2. pass 1. 90%.
2. pass 1. 87%.
2. pass 1. 91%.
2. pass biological activity
(HUVEC cell proliferation inhibitory ability) IC50 value is within 70 - 130% of standard product 91% 96% 86% 84%

As can be seen in Table 2, the syringe formulation of the present invention was found to maintain the same physical properties as in the initial stage even under refrigerated storage conditions and storage conditions for up to 6 months, and biological activity was found to be superior to that of the initial stage.

In addition, as can be seen in Table 3 above, it was confirmed that the syringe formulation of the present invention had physical properties at a level almost similar to that of the initial stage even during long-term storage under accelerated storage conditions. Although the biological activity was lowered after storage for 6 months compared to the initial stage, it was confirmed that it was at a level that meets the criteria for use as a pharmaceutical preparation.

According to these results, the syringe containing the ophthalmic formulation of the present invention does not generate foreign substances even when stored for a long time in refrigerated storage and accelerated storage conditions, and has the advantage of maintaining excellent physical properties as the initial ophthalmic formulation. In particular, despite the result of maintaining the body in a silicone oil-free condition, the result was maintained without significant change in terms of protein content.

Example 4: Confirmation of effect of reducing drug contamination of blister packaged products by coupling of stopper and plunger

Prepare a syringe manufactured by the method of Example 2, on the other hand, the other components of the syringe were prepared in the same manner as in Example 2, but there was no protrusion at the distal end of the plunger rod, so the stopper and the plunger were not coupled separately. After manufacturing, each of the two products was packaged in the form of Tyvek sealed blisters.

In order to further improve product safety when injecting into the vitreous for ophthalmology in the future, using a sterilizer of the VHP LTS-V model (Steris), steam hydrogen peroxide inside the blister pack is used for the same conditions for both products. sterilization process was performed. At this time, using a vacuum pump at a temperature of 28 ℃ so that the vapor hydrogen peroxide gas passes through the Tyvek membrane and effectively penetrates into the blister pack, the pressure is reduced to about 10 mbar, and then the process of injecting the vapor hydrogen peroxide is performed for a total of 7 cycles. proceeded. It was confirmed that the inside of the blister pack (outside of the syringe) was effectively sterilized for both products through the biomarker that can confirm sterilization.

However, in the case of a syringe in which the stopper and the plunger are not coupled, microbial contamination of the chemical solution filled in the syringe in some products could not be avoided. This causes the stopper to retreat as the air bubbles generated in the chemical filling process swell under reduced pressure condition, and the distance at which the stopper retreated exceeds the distance (4mm) where the stopper and the barrel were joined. Because it was moved to the location. It is interpreted that the chemical solution in contact with the inner surface of the barrel where the sterility is not maintained does not exert the sterilization effect because the steam hydrogen peroxide does not penetrate to the chemical layer even in the steam hydrogen peroxide sterilization process.

On the other hand, in the case of the product in which the stopper and the plunger are coupled, the movement distance of the stopper is relatively small even under reduced pressure, so that microbial contamination does not occur at all. Table 6 below shows the average moving distance and the rate of microbial contamination of the stoppers in the two products.

sample group Stopper retraction average moving distance (mm) Microbial contamination rate (%) Separable product with stopper and plunger rod 4.6 25 Combination of stopper and plunger rod 2.8 0

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims to be described later and their equivalents.

1: syringe
2: body
4: plunger
6: Reverse prevention part
8: seal
A: 1st axis
10: stopper
12: Exit
14: exit end
16: front surface
18: variable volume chamber
20: ophthalmic formulation
22: flange portion
23: insertion fixture
24: plunger first end
25: rear
26: plunger rod
28: end flange
30: sandwich portion
32: plunger shoulder
34: anti-reverse shoulder
36: bore
38: rear end
40: finger protrusion
42: disk shaped flange
44: plunger rib
46: disk-shaped part
48, 50: gap
52, 56: ribs
58: stopper body
60: rear surface
62: insertion fixing groove
64: bore having a first diameter
66: internal fixing groove having a second diameter
64': the lower end of the spherical insertion fixing part
66': the upper part of the spherical insertion fixing part
81: upper part
82: lower part
83: depression
84: protrusion

<110> SAM CHUN DANG PHARM. CO., LTD. <120> Syringe comprising ophthalmic formulation <130> SCD116 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 431 <212> PRT <213> Artificial Sequence <220> <223> Aflibercept <400> 1 Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu 1 5 10 15 Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val 20 25 30 Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr 35 40 45 Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe 50 55 60 Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu 65 70 75 80 Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg 85 90 95 Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly Ile 100 105 110 Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg Thr 115 120 125 Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys 130 135 140 His Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser Gly 145 150 155 160 Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val Thr 165 170 175 Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met 180 185 190 Thr Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys Asp Lys Thr 195 200 205 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 210 215 220 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 225 230 235 240 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 245 250 255 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 260 265 270 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 275 280 285 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 290 295 300 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 305 310 315 320 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 325 330 335 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 340 345 350 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 355 360 365 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 370 375 380 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 385 390 395 400 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 405 410 415 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 420 425 430

Claims (20)

a body (2) having an outlet (12) at the outlet end (14);
a plunger (4) located inside the body (2); and
Located in the interior of the body (2) at the front of the plunger (4), it is silicon oil free and has a front surface (16) forming a variable volume chamber (18) with the body (2). and a stopper 10 for discharging the ophthalmic formulation 20 filled in the variable volume chamber 18 through the outlet 12 by the movement of the plunger 4,
The plunger (4) includes a plunger rod (26) extending in the longitudinal direction, and a flange portion (22) located at a first end (24) of the plunger rod (26) adjacent to the stopper (10); Includes a spherical or screw-shaped insertion fixing part 23 protruding from the flange part 22 toward the stopper 10,
The stopper 10 is provided with an insertion fixing groove 62 into which the insertion fixing part 23 is inserted and fixed, and the ophthalmic formulation is in an aqueous medium, (a) 10 to 100 mg in a therapeutically effective amount. aflibercept in /ml; (b) 2 to 12 w/v% of a stabilizer; (c) 0.01 to 0.1 w/v% of a surfactant; and (d) 5-100 mM of an ionic isotonic agent; Including, having a pH of 5.2 to 6.2, silicone-free (silicon oil free) syringe. According to claim 1,
A syringe in which the insertion fixing part 23 is fixed to the insertion fixing groove 62 of the stopper 10 by press-fitting. According to claim 1,
A syringe having a spherical shape in which the insertion fixing part 23 protrudes toward the stopper 10 . According to claim 1,
The syringe is made of plastic, and the body is silicone-free. 5. The method of claim 4,
The syringe, wherein the plastic is selected from the group consisting of cycloolefin polymers (COP) and cycloolefin copolymers (COC). According to claim 1,
the plunger (4) comprises at least one plunger shoulder (32) located on the part of the plunger rod (26) opposite the first end (24);
In the rear part 25 of the body 2, when the plunger 4 moves backward, it includes a shoulder 34 for preventing the plunger from being separated by cooperation with the plunger shoulder 32. Syringe that is further provided with a reverse prevention unit (6). 7. The method of claim 6,
The flange portion 22 is arranged such that the plunger contact portion 22 is coupled with a stopper 10 and the plunger shoulder 32 and the anti-retraction shoulder 34 are at full volume when the variable volume chamber 18 is at its maximum volume. Syringes with a gap of 2 mm or less between them. According to claim 1,
A syringe wherein said outlet is reversibly sealed. According to claim 1,
A syringe for use in administering an ophthalmic formulation to a patient with an ophthalmic condition. According to claim 1,
The syringe, wherein a therapeutically effective amount of aflibercept has the amino acid sequence consisting of SEQ ID NO: 1. According to claim 1,
The ophthalmic formulation further comprises a buffer of 5 to 20 mM, a syringe. 12. The method of claim 11,
The buffer solution is an acetate buffer solution, syringe. 12. The method of claim 11,
The buffer solution is sodium acetate, syringe. According to claim 1,
The stabilizer is any one selected from the group consisting of sucrose, trehalose, mannitol and glucose syringe. According to claim 1,
The stabilizer is a syringe, which is sucrose. According to claim 1,
The syringe, wherein the surfactant is at least one selected from the group consisting of polysorbate 20, polysorbate 40, polysorbate 60 and polysorbate 80. According to claim 1,
The surfactant is polysorbate 20 syringe. According to claim 1,
Syringe in which the ionic isotonic agent is sodium chloride. According to claim 1,
The ophthalmic formulation comprises (a) a therapeutically effective amount of 20-50 mg/ml of aflibercept; (b) 8-12 mM sodium acetate buffer; (c) 5 to 10 w/v% sucrose; (d) 0.01 to 0.05 w/v% polysorbate 20; and (e) 15-40 mM of an ionic isotonic agent of sodium chloride; Including, and having a pH of 5.4 to 6.0, a syringe. 1) preparing the ophthalmic formulation of claim 1;
2) sterilizing and filtering the prepared ophthalmic formulation;
3) aseptically filling the sterilized and filtered formulation into the variable volume chamber of a syringe using a pump of an automatic filling machine; and
4) aseptically mounting the stopper by an automatic charger; and
5) coupling the stopper with the plunger; a method for filling an ophthalmic formulation in a syringe comprising a.

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